Comparative studies on the 5-aminolaevulinic acid dehydratases from Pisum sativum, Escherichia coli and Saccharomyces cerevisiae.
5-Aminolaevulinic acid dehydratase (ALAD) is an essential enzyme in most organisms, catalysing an inaugural step in the tetrapyrrole biosynthetic pathway, the Knorr-type condensation reaction of two molecules of 5-aminolaevulinic acid (ALA) to form the monopyrrole porphobilinogen. ALADs can be conveniently separated into two main groups: those requiring Zn2+ for activity (typified here by the enzymes from Escherichia coli and Saccharomyces cerevisiae, yeast) and those requiring Mg2+ (represented here by the enzyme from Pisum sativum, pea). Here we describe a detailed comparison of these two metal-dependent systems. Kinetically influential ionizations were identified by using pH-dependent kinetics. Groups with pKa values of approx. 7 and 10 (assigned to cysteine and lysine residues) were detected in the free enzyme and enzyme-substrate states of all three enzymes, and a further ionizable group with a pKa of approx. 8.5 (assigned to histidine) was found to be additionally important to the yeast enzyme. The importance of these residues was confirmed by using protein modifying reagents. Shifts in the pKa values of the pea and E. coli enzymes consequent on E-S complex formation suggest a change to a less hydrophobic micro-environment when substrate binds. Studies with inhibitors revealed that the three enzymes exhibit differential susceptibilities and, in the case of succinylacetone, this is reflected in Ki values that vary by three orders of magnitude. In addition, the crystallization of the yeast ALAD is described, raising the possibility of an X-ray-derived three-dimensional structure of this enzyme.


Impact of perinatal factors on continuous early monitoring of brain electrocortical activity in very preterm newborns by amplitude-integrated EEG.
Amplitude-integrated electroencephalogram (aEEG) is increasingly used for neuromonitoring in preterms. We aimed to quantify the effects of gestational age (GA), postnatal age (PNA), and other perinatal factors on the development of aEEG early after birth in very preterm newborns with normal cerebral ultrasounds. Continuous aEEG was prospectively performed in 96 newborns (mean GA: 29.5 (range: 24.4-31.9) wk, birth weight 1,260 (580-2,120) g) during the first 96 h of life. aEEG tracings were qualitatively (maturity scores) and quantitatively (amplitudes) evaluated using preestablished criteria. A significant increase in all aEEG measures was observed between day 1 and day 4 and for increasing GA (P < 0.001). The effect of PNA on aEEG development was 6.4- to 11.3-fold higher than that of GA. In multivariate regression, GA and PNA were associated with increased qualitative and quantitative aEEG measures, whereas small-for-GA status was independently associated with increased maximum aEEG amplitude (P = 0.003). Morphine administration negatively affected all aEEG measures (P < .05), and caffeine administration negatively affected qualitative aEEG measures (P = 0.02). During the first few days after birth, aEEG activity in very preterm infants significantly develops and is strongly subjected to the effect of PNA. Perinatal factors may alter the early aEEG tracing and interfere with its interpretation.


The effect of school meals with fatty fish on adolescents' self-reported symptoms for mental health: FINS-TEENS - a randomized controlled intervention trial.
There is a growing body of evidence linking fish consumption and n-3 LCPUFAs to mental health. Still, the results from randomized trials with n-3 LCPUFAs show conflicting results, and it is possible that the combined effect of several nutrients in fish may explain the observed associations. To aim of the present study was to investigate if school meals with fatty fish three times per week for 12 weeks could alter mental health in a sample of typically developing adolescents. In the Fish Intervention Studies-TEENS (FINS-TEENS), adolescents from eight secondary schools (n=425) in Norway, were randomized to receive school meals with fatty fish, meat or n-3 LCPUFA supplements. Mental health was assessed with the Strengths and Difficulties Questionnaire (SDQ) and the differences between the groups were assessed with linear mixed effect models, unadjusted and adjusted for baseline and dietary compliance. The results showed no effects of school meals with fatty fish compared to similar meals with meat or n-3 LCPUFAs on the adolescents' self-reported symptom scores for mental health. Among adolescents scoring above the SDQ cut-offs (high-scorers), the fish- improved less than the meat group in the self-reported symptom scores for total difficulties- and emotional problems. However, the findings should be regarded as preliminary, as the analyses for the high-scorer group were underpowered. In conclusion, serving school meals with fatty fish did not alter mental health in a typically developing sample of adolescents. It is possible that serving healthy school meals with meat is more beneficial than similar meals with fatty fish in adolescents scoring high on mental health problems. However, the results should be seen as preliminary, as the dietary compliance in the fish group was low and the analyses in the high score group underpowered. Thus, further studies should investigate the associations between fish consumption and adolescents' mental health.


Gastrointestinal motility disorders in neurologic disease.
The extrinsic and autonomic nervous system intricately controls the major functions of the gastrointestinal tract through the enteric nervous system; these include motor, secretory, sensory, storage, and excretory functions. Disorders of the nervous system affecting gastrointestinal tract function manifest primarily as abnormalities in motor (rather than secretory) functions. Common gastrointestinal symptoms in neurologic disorders include sialorrhea, dysphagia, gastroparesis, intestinal pseudo-obstruction, constipation, diarrhea, and fecal incontinence. Diseases of the entire neural axis ranging from the cerebral hemispheres to the peripheral autonomic nerves can result in gastrointestinal motility disorders. The most common neurologic diseases affecting gastrointestinal function are stroke, parkinsonism, multiple sclerosis, and diabetic neuropathy. Diagnosis involves identification of the neurologic disease and its distribution, and documentation of segmental gut dysfunction, typically using noninvasive imaging, transit measurements, or intraluminal measurements of pressure activity and coordination of motility. Apart from treatment of the underlying neurologic disease, management focuses on restoration of normal hydration and nutrition and pharmacologic treatment of the gut neuromuscular disorder.


DR (Ia-like) antigens on human melanoma cells. Serological detection and immunochemical characterization.
11 cultured human melanoma cell lines were tested for the expression of DR antigens by using specific allo- and xenoantisera in an indirect rosette microassay. Four of these melanoma cell lines expressed DR antigens, but in lower amounts than expressed on cultured human B-lymphoid cells. Rabbits injected with the DR-positive melanoma cells produced antibodies that were serologically and immunochemically reactive with B-cell-derived DR antigens. Immunochemical studies indicate that melanoma cell-derived DR antigens have a two-chain structure with 34,000 and 27,000 mol wt components. The melanoma cell-derived DR beta-chain at 27,000 mol wt is slightly smaller than that of the Victor cell DR beta-chain whose mol wt is 29,000.


Trends in Comorbidities and Complications Among Patients Undergoing Hip Fracture Repair.
Hip fracture patients represent various perioperative challenges related to their significant comorbidity burden and the high incidence of morbidity and mortality. As population trend data remain rare, we aimed to investigate nationwide trends in the United States in patient demographics and outcomes in patients after hip fracture repair surgery. After Institutional Review Board (IRB) approval (IRB#2012-050), data covering hip fracture repair surgeries were extracted from the Premier Healthcare Database (2006-2016). Patient demographics, comorbidities, and complications, as well as anesthesia and surgical details, were analyzed over time. Cochran-Armitage trend tests and simple linear regression assessed significance of (linear) trends. Among N = 507,274 hip fracture cases, we observed significant increases in the incidence in preexisting comorbid conditions, particularly the proportion of patients with >3 comorbid conditions (33.9% to 43.4%, respectively; P < .0001). The greatest increase for individual comorbidities was seen for sleep apnea, drug abuse, weight loss, and obesity. Regarding complications, increased rates over time were seen for acute renal failure (from 6.9 to 11.1 per 1000 inpatient days; P < .0001), while significant decreasing trends for mortality, pneumonia, hemorrhage/hematoma, and acute myocardial infarction were recorded. In addition, decreasing trends were observed for the use of neuraxial anesthesia either used as sole anesthetic or combined with general anesthesia (7.3% to 3.6% and 6.3% to 3.4%, respectively; P < .0001). Significantly more patients (31.9% vs 41.3%; P < .0001) were operated on in small rather than medium- and large-sized hospitals. From 2006 to 2016, the overall comorbidity burden increased among patients undergoing hip fracture repair surgery. Throughout this same time period, incidence of postoperative complications either remained constant or declined with the only significant increase observed in acute renal failure. Moreover, use of regional anesthesia decreased over time. This more comorbid patient population represents an increasing burden on the health care system; however, existing preventative measures appear to be effective in minimizing complication rates. Although, given the proposed benefits of regional anesthesia, decreased utilization may be of concern.


CRISPR-Cas13d Exhibits Robust Antiviral Activity Against Seneca Valley Virus.
In recent years, Seneca Valley virus (SVV) as a newly identified pathogen of porcine vesicular disease spread quickly and has posed a potential threat to the swine industry in several countries resulting in economic losses. Considering the evolution of SVV, attention should be given to controlling SVV epidemics. So far there are no commercial vaccines or drugs available to combat SVV. Therefore, development of strategies for preventing and controlling SVV infection should be taken into account. In the current study, we evaluated whether the CRISPR-Cas13d system could be used as a powerful tool against SVV infection. Besides, selected crRNAs showed different capacity against SVV infection. Our study suggests the CRISPR-Cas13d system significantly inhibited SVV replication and exhibited potent anti-SVV activity. This knowledge may provide a novel alternative strategy to control epidemics of SVV in the future.


Diagnosis of rinderpest in Tanzania by a rapid chromatographic strip-test.
A simple chromatographic strip-test based on Clearview technology, is under development as a pen-side test for the detection of rinderpest antigen in eye swabs taken from cattle in the field. An outbreak of rinderpest occurred in the northern zone of Tanzania from late February to June 1997. The affected cattle exhibited very mild clinical signs, which made clinical diagnosis difficult. One hundred and seven eye swabs were collected from cattle suspected of infection with rinderpest. These were tested in the field using a prototype of the pen-side test and 13 (12.15%) of the samples were found to be positive for the presence of rinderpest antigen. These were confirmed by ICE. The positive cases were predominantly found in the Ngorongoro district. This demonstrates the usefulness of such a simple, rapid pen-side diagnostic assay, particularly when clinically 'mild' strains of rinderpest are present.


Image processing pipeline for segmentation and material classification based on multispectral high dynamic range polarimetric images.
We propose a method for the capture of high dynamic range (HDR), multispectral (MS), polarimetric (Pol) images of indoor scenes using a liquid crystal tunable filter (LCTF). We have included the adaptive exposure estimation (AEE) method to fully automatize the capturing process. We also propose a pre-processing method which can be applied for the registration of HDR images after they are already built as the result of combining different low dynamic range (LDR) images. This method is applied to ensure a correct alignment of the different polarization HDR images for each spectral band. We have focused our efforts in two main applications: object segmentation and classification into metal and dielectric classes. We have simplified the segmentation using mean shift combined with cluster averaging and region merging techniques. We compare the performance of our segmentation with that of Ncut and Watershed methods. For the classification task, we propose to use information not only in the highlight regions but also in their surrounding area, extracted from the degree of linear polarization (DoLP) maps. We present experimental results which proof that the proposed image processing pipeline outperforms previous techniques developed specifically for MSHDRPol image cubes.


3H-1,2-Dithiole-3-thione as a novel therapeutic agent for the treatment of ischemic stroke through Nrf2 defense pathway.
Cerebral ischemic stroke accounts for more than 80% of all stroke cases. During cerebral ischemia, reactive oxygen species produced in brain tissue induce oxidative stress and inflammatory responses. D3T, the simplest compound of the cyclic, sulfur-containing dithiolethiones, is found in cruciferous vegetables and has been reported to induce antioxidant genes and glutathione biosynthesis through activation of Nrf2. In addition to antioxidant activity, D3T was also reported to possess anti-inflammatory effects. In this study, we evaluated the therapeutic potential of D3T for the treatment of ischemic stroke and investigated the mechanisms underlying the protective effects of D3T in ischemic stroke. Mice subjected to transient middle cerebral artery occlusion/reperfusion (tMCAO/R) were administered with vehicle or D3T to evaluate the effect of D3T in cerebral brain injury. We observed D3T reduced infarct size, decreased brain edema, lessened blood-brain barrier disruption, and ameliorated neurological deficits. Further investigation revealed D3T suppressed microglia (MG) activation and inhibited peripheral inflammatory immune cell infiltration of CNS in the ischemic brain. The protective effect of D3T in ischemic stroke is mediated through Nrf2 induction as D3T-attenuated brain injury was abolished in Nrf2 deficient mice subjected to tMCAO/R. In addition, in vitro results indicate the induction of Nrf2 by D3T is required for its suppressive effect on MG activation and cytokine production. In summary, we demonstrate for the first time that D3T confers protection against ischemic stroke, which is mediated through suppression of MG activation and inhibition of CNS peripheral cell infiltration, and that the protective effect of D3T in ischemic stroke is dependent on the activation of Nrf2.


Tunable vapor-condensed nanolenses.
Nanostructured optical components, such as nanolenses, direct light at subwavelength scales to enable, among others, high-resolution lithography, miniaturization of photonic circuits, and nanoscopic imaging of biostructures. A major challenge in fabricating nanolenses is the appropriate positioning of the lens with respect to the sample while simultaneously ensuring it adopts the optimal size and shape for the intended use. One application of particular interest is the enhancement of contrast and signal-to-noise ratio in the imaging of nanoscale objects, especially over wide fields-of-view (FOVs), which typically come with limited resolution and sensitivity for imaging nano-objects. Here we present a self-assembly method for fabricating time- and temperature-tunable nanolenses based on the condensation of a polymeric liquid around a nanoparticle, which we apply to the high-throughput on-chip detection of spheroids smaller than 40 nm, rod-shaped particles with diameter smaller than 20 nm, and biofunctionalized nanoparticles, all across an ultralarge FOV of >20 mm(2). Previous nanoparticle imaging efforts across similar FOVs have detected spheroids no smaller than 100 nm, and therefore our results demonstrate the detection of particles >15-fold smaller in volume, which in free space have >240 times weaker Rayleigh scattering compared to the particle sizes detected in earlier wide-field imaging work. This entire platform, with its tunable nanolens condensation and wide-field imaging functions, is also miniaturized into a cost-effective and portable device, which might be especially important for field use, mobile sensing, and diagnostics applications, including, for example, the measurement of viral load in bodily fluids.


Oral squamous cell carcinoma arising from areas of Graft-versus-host disease: A systematic review.
Graft-versus-host disease (GVHD) is an immune system reaction that occurs in patients with a history of hematopoietic stem cell transplantation (HSCT), in which the grafted donor's cells attack those of the host. The objective of this systematic review was to present a study on oral squamous cell carcinoma (OSSC) that developed from GVHD areas in patients undergoing HSCT. An electronic search was conducted in the databases PUBMED, WEB OF SCIENCE, SCOPUS, MEDLINE and SCIENCE DIRECT, according to PRISMA guidelines. Of the 1582 results, 23 articles were included, resulting in 81 cases. The most common underlying disease for performing the transplant was Myeloid Leukemia (55.6%). The mean age was 39 years, with a predilection for males (64.2%). The tongue was the site of GVHD that most frequently underwent transformation to SCC (59.3%). The average time between transplantation and the development of GVHD was of approximately of 8 months, while the average period of development between transplantation and the development of OSCC was of approximately of 111 months. The most common treatment to GVHD was cyclosporine associated with corticosteroids. OSCCs arising from areas of GVHD present a different evolution from conventional oral carcinomas, since they affect younger patients, smoking and alcohol are not important etiological factors and finally because they present good prognosis, but further studies with larger number cases followed are needed to confirm our findings.


Multidetector CT appearance of the pelvis after vaginal delivery: normal appearances and abnormal acute findings.
Vaginal delivery is the most commonly performed delivery in the world and accounts for nearly two-thirds of all deliveries in the United States. It is a secure method but may be associated with some acute complications, especially in the immediate postpartum days, which can potentially be fatal for the mother. The most frequent acute complications are hemorrhages/hematomas, uterine rupture, endometritis, retained product of conception (RPOC), ovarian thrombosis and HELLP syndrome (hemolysis, elevated liver enzymes, low platelet count). A first evaluation of the clinical status of the patients is executed by the clinicians who, depending on their experience, perform ultrasonography by themselves and eventually may request further radiologic exams in doubtful cases. Radiologists may play an important role recognizing early postpartum complications and differentiating them from physiologic postoperative findings. In this setting, the use of multidetector computed tomography (MDCT) is important for diagnosis of suspected postpartum complications. The aim of this article is to review the normal and abnormal post vaginal delivery MDCT aspects in order to help the clinical management by preventing misdiagnoses and tailoring the best medical treatments.


Gene therapy of spontaneous canine melanoma and feline fibrosarcoma by intratumoral administration of histoincompatible cells expressing human interleukin-2.
The production of human interleukin-2 (hIL-2) local to the tumor site by engineered histoincompatible cells has been shown in various murine models to promote a strong immune response leading to tumor growth inhibition or rejection. To assess whether this strategy would be similarly applicable for treatment of primary neoplastic cells, two naturally occurring tumors were used as preclinical models; the highly metastatic melanoma of the dog and the low metastatic fibrosarcoma of the cat. We demonstrate that both cats and dogs when treated by tumor surgery, radiotherapy and repeated local injections of xenogeneic Vero cells secreting high levels of hIL-2 relapse less frequently and survive longer than control animals treated by surgery and radiotherapy alone. Local secretion of hIL-2 by the xenogeneic cells is shown to be necessary for the induction of an optimal antitumor effect. Moreover, the safety of the procedure was demonstrated in both animal models and through extensive toxicological analysis performed in rats. These results confirm for the first time to our knowledge the safety and therapeutic potential of a gene transfer strategy in animals with spontaneous metastatic and nonmetastatic tumors.


Recognition memory for new associations: electrophysiological evidence for the role of recollection.
The electrophysiological correlates of recognition memory for new associations were investigated in two experiments. In both experiments subjects first studied unrelated word pairs. At test, they were presented with old words in the same pairing as at study (same pairs), old words in a different pairing from study (rearranged pairs), and pairs of new words. In Experiment 1 the test requirement was to discriminate between old and new pairs and, for any pair judged old, to then judge whether the pair was the same or rearranged. In Experiment 2 the requirement was merely to discriminate between old and new pairs. Event-related potentials (ERPs) were recorded for correctly classified same, rearranged and new pairs. The ERPs elicited by same pairs exhibited a similar pattern of effects in both experiments. Relative to the ERPs to new pairs, these effects took the form of sustained positive shifts with two distinct scalp maxima, over the left temporo-parietal and right frontal scalp respectively. ERPs to rearranged pairs showed effects which were similar in scalp topography, but markedly smaller in magnitude. This pattern of ERP effects closely resembles that found previously for test items defined as recollected on the basis of their attracting a successful source judgement. The findings therefore suggest that associative recognition memory shares some of the recollective processes that are engaged by the requirement to retrieve contextual information about a study episode. The findings from Experiment 2 indicate that the processes associated with the recollection of associated pairs are engaged regardless of whether the retrieval of associative information is an explicit task requirement.


Protein phosphatase 1 regulates huntingtin exon 1 aggregation and toxicity.
Huntington's disease is neurodegenerative disorder caused by a polyglutamine expansion in the N-terminal region of the huntingtin protein (N17). Here, we analysed the relative contribution of each phosphorylatable residue in the N17 region (T3, S13 and S16) towards huntingtin exon 1 (HTTex1) oligomerization, aggregation and toxicity in human cells and Drosophila neurons. We used bimolecular fluorescence complementation to show that expression of single phosphomimic mutations completely abolished HTTex1 aggregation in human cells. In Drosophila, mimicking phosphorylation at T3 decreased HTTex1 aggregation both in larvae and adult flies. Interestingly, pharmacological or genetic inhibition of protein phosphatase 1 (PP1) prevented HTTex1 aggregation in both human cells and Drosophila while increasing neurotoxicity in flies. Our findings suggest that PP1 modulates HTTex1 aggregation by regulating phosphorylation on T3. In summary, our study suggests that modulation of HTTex1 single phosphorylation events by PP1 could constitute an efficient and direct molecular target for therapeutic interventions in Huntington's disease.


Modeling multi-decadal mangrove leaf area index in response to drought along the semi-arid southern coasts of Iran.
Leaf Area Index (LAI; as an indicator of the health) of the mangrove ecosystems on the northern coasts of the Persian Gulf and the Gulf of Oman was measured in the field and modeled in response to observed (1986-2017) and predicted (2018-2100) drought occurrences (quantified using the Standardized Precipitation Index [SPI]). The relationship of LAI with the normalized difference vegetation index (NDVI) obtained from satellite images was quantified, the LAI between 1986 and 2017 retrospectively estimated, and a relationship between LAI and SPI developed for the same period. Long-term climate data were used as input in the RCP8.5 climate change scenario to reconstruct recent and forecast future drought intensities. Both the NDVI and the SPI were strongly related with the LAI, indicating that realistic LAI values were derived from historic satellite data to portray annual changes of LAI in response to changes in SPI. Our findings show that projected future drought intensities modeled by the RCP8.5 scenario increase more and future LAIs decreased more on the coasts of the Gulf of Oman than the coasts of the Persian Gulf in the coming decades. The year 1998 was the most significant change-point for mean annual rainfall amounts and drought occurrences as well as for LAIs and at no time between 1998 and 2017 or between 2018 and 2100 are SPI and LAI values expected to return to pre-1998 values. LAI and SPI are projected to decline sharply around 2030, reach their lowest levels between 2040 and 2070, and increase and stabilize during the late decades of the 21st century at values similar to the present time. Overall, this study provides a comprehensive picture of the responses of mangroves to fluctuating future drought conditions, facilitating the development of management plans for these vulnerable habitats in the face of future climate change.


A biomechanical comparison of conventional classifications of bowling action-types in junior fast bowlers.
Fast bowling is categorised into four action types: side-on, front-on, semi-open and mixed; however, little biomechanical comparison exists between action types in junior fast bowlers. This study investigated whether there are significant differences between action-type mechanics in junior fast bowlers. Three-dimensional kinematic and kinetic analyses were completed on 60 junior male fast bowlers bowling a five-over spell. Mixed-design factorial analyses of variance were used to test for differences between action-type groups across the phases of the bowling action. One kinetic difference was observed between groups, with a higher vertical ground reaction force loading rate during the front-foot contact phase in mixed and front-on compared to semi-open bowlers; no other significant group differences in joint loading occurred. Significant kinematic differences were observed between the front-on, semi-open and mixed action types during the front-foot contact phase for the elbow and trunk. Significant kinematic differences were also present for the ankle, T12-L1, elbow, trunk and pelvis during the back-foot phase. Overall, most differences in action types for junior fast bowlers occurred during the back-foot contact phase, particularly trunk rotation and T12-L1 joint angles/ranges of motion, where after similar movement patterns were utilized across groups during the front-foot contact phase.


Complex Scalp and Calvarium Defects After Giant Basal Cell Carcinoma Excision: Management, Challanges, Outcomes.
Giant basal cell carcinoma (GBCC) is defined as a tumor ≥5 cm in diameter. GBCC of scalp usually requires extended resection of soft tissues, calvarium, and dura. In this study, we present 5 patients with GBCC of head, who underwent a single-stage combined scalp, calvarium, and dural reconstruction. Herein, we aim to discuss reconstruction methods, cerebrospinal fluid (CSF) leakage, duration of hospital stay, and tumor recurrency. Peroperative and postoperative follow-ups, defect areas, and performed calvarium reconstruction methods of 5 patients, who underwent complex scalp and calvarium reconstruction after GBCC between year 2010 and 2017, were retrospectively maintained. We studied reconstruction methods, CSF leakage, duration of hospital stay, and tumor recurrency. All patients undergone single-stage reconstruction. Avarge duration of hospital stay was 15 days. Titanium mesh was used in 3 patients and methyl methacrylate was used in 2 patients for calvarium reconstruction. CSF leakage was seen in patients who underwent calvarium reconstruction with titanium mesh. Tumor recurrence occured next to calvarium in 1 patient who undergone calvarium reconstruction with methyl methacrylate. CSF leakage and duration of hospital stay may induce morbidity of this oncoplastic procedure. The fact of longer hospital stay of patients reconstructed with titanium mesh might be a new data presented in this study. These parameters can be related with the method of calvarium reconstruction.


Genetic crosses in the apicomplexan parasite Cryptosporidium parvum define recombination parameters.
Recombinant progeny lines of Cryptosporidium parvum were generated by coinfecting immunosuppressed mice with two genetically distinct isolates of C. parvum. Progeny lines were obtained from a cross of parental lines MD x TU114 through targeted propagation in mice of progeny oocysts originating from populations lacking one parental allele at one or more loci. For each infection lineage this process was repeated until only a single allele remained for each marker, indicating that the progeny line was clonal. To study genetic recombination, 16 progeny clones were genotyped at 40 loci located on each of the eight chromosomes. The inheritance of parental alleles was significantly skewed towards the more virulent parent isolate MD. A contiguous 476 kb segment of chromosome V displayed MD allele in all progeny recovered, while MD and TU114 alleles were detected at other loci throughout the genome. The absence of alleles from one parental isolate in this chromosomal region may indicate phenotypic selection for the MD allele during the generation of these lines. A range for the meiotic crossover frequency was determined on the basis of 40 markers and the number of meioses estimated to have taken place during the crossing experiment. C. parvum exhibits a high rate of recombination commensurate with other Apicomplexa.


Protective effect of Spirulina platensis against cell damage and apoptosis in hepatic tissue caused by high fat diet.
Spirulina platensis is a microalga that may be a source of antioxidants that can reduce body fat deposition. Consumption of a high fat diet produces elevated blood lipid levels, inflammation and apoptosis. We investigated the possible effects of S. platensis on the blood lipid profile, and liver inflammation and apoptosis in rats fed a high fat diet. Sixty-four young male rats were divided into eight equal groups. The control group was fed a basic diet. The experimental groups were fed a diet for 60 days that was prepared by mixing variable amounts of 43% vegetable oil and 10% cholesterol with or without 3% S. platensis mixed with the basal diet. Blood and liver tissue samples were collected from each animal. Serum samples were used to analyze lipid parameters, total antioxidant status and total oxidant status. iNOS and eNOS were determined by immunohistochemistry. TUNEL staining was used to detect apoptosis to investigate a possible connection between inflammation and apoptosis in the liver tissue. The relations between fat deposition and liver degeneration were assessed by Sirius red staining and alpha-smooth muscle actin immunostaining. S. platensis reduced serum HDL-C, LDL-C and triglyceride, increased HDL-C levels in rats fed a high fat diet to near control levels, and reduced iNOS levels and increased eNOS levels in the liver tissue compared to vegetable oil and cholesterol treated groups. The apoptotic index was reduced in the groups that were fed a high fat or a basic diet when supplemented with S. platensis.


A Case of Successful Therapy by Intrapleural Injection of Fibrin Glue for Chylothorax after Lung Transplantation for Lymphangioleiomyomatosis.
A 37-year-old woman underwent bilateral lung transplantation for lymphangioleiomyomatosis. Dense pleural adhesions due to past pleurodesis for chylothorax were observed and dissected in both thoracic cavities. The patient developed chylothorax after transplant. Chylothorax in the right thoracic cavity was successfully treated by conventional pleurodesis; however, pleural effusion from the left thoracic cavity was not reduced. According to fluoroscopic images obtained by injecting a contrast medium through the chest tube, the remaining pleural space in the left thoracic cavity was small and localized in the lower region adjacent to the mediastinum. We opted to fill this space with fibrin glue; we injected fibrinogen and thrombin solution into the space through the chest tube. We performed fibrin glue treatment three times and pleural effusion was dramatically decreased. We removed the chest tube on day 107 post-transplant. No recurrent chylothorax has been recorded for 10 years after lung transplantation.


Interrater and intrarater reliability of finger goniometric measurements.
Intrarater and interrater reliability were determined for middle finger range of motion (ROM) using the Rolyan finger goniometer. Seven raters measured ROM 3 times (trials) on 20 healthy adults. Intraclass reliability analysis and analysis of variance was used to assess the consistency and stability of measures. Level of significance was set at p < or = .05. Significant differences (p < .05) existed between raters for metacarpophalangeal, proximal interphalangeal, and distal interphalangeal active finger ROM measurements and for metacarpophalangeal and proximal interphalangeal passive range of motion. Intrarater reliability ranged from .43 to .99 (single measure intraclass coefficient). Interrater reliability ranged from .24 to .95, with passive measures lower than active measures. These results provide clinicians with evidence for using this specific goniometer and issues associated with various therapists assessing finger ROM.


Improved catalytic efficiency and active site modification of 1,4-beta-D-glucan glucohydrolase A from Thermotoga neapolitana by directed evolution.
Thermotoga neapolitana 1,4-beta-d-glucan glucohydrolase A preferentially hydrolyzes cello-oligomers, such as cellotetraose, releasing single glucose moieties from the reducing end of the cello-oligosaccharide chain. Using directed evolution techniques of error-prone PCR and mutant library screening, a variant glucan glucohydrolase has been isolated that hydrolyzes the disaccharide, cellobiose, at a 31% greater rate than its wild type (WT) predecessor. The mutant library, expressed in Escherichia coli, was screened at 85 degrees C for increased hydrolysis of cellobiose, a native substrate rather than a chromogenic analog, using a continuous, thermostable coupled enzyme assay. The V(max) for the mutant was 108 +/- 3 units mg(-1), whereas that of the WT was 75 +/- 2 units mg(-1). The K(m) for both proteins was nearly the same. The k(cat) for the new enzyme increased by 31% and its catalytic efficiency (k(cat)/K(m)) for cellobiose also rose by 31% as compared with the parent. The nucleotide sequence of two positive clones and two null clones identified 11 single base shifts. The nucleotide transition in the most active clone caused an isoleucine to threonine amino acid substitution at position 170. Structural models for I170T and WT proteins were derived by sequence homology with Protein Data Bank code 1BGA from Paenibacillus polymyxa. Analysis of the WT and I170T model structures indicated that the substitution in the mutant enzyme repositioned the conserved catalytic residue Asn-163 and reconfigured entry to the active site.


Effectiveness of Chiropractic Care to Improve Sensorimotor Function Associated With Falls Risk in Older People: A Randomized Controlled Trial.
This study assessed whether 12 weeks of chiropractic care was effective in improving sensorimotor function associated with fall risk, compared with no intervention, in community-dwelling older adults living in Auckland, New Zealand. Sixty community-dwelling adults older than 65 years were enrolled in the study. Outcome measures were assessed at baseline, 4 weeks, and 12 weeks and included proprioception (ankle joint position sense), postural stability (static posturography), sensorimotor function (choice stepping reaction time), multisensory integration (sound-induced flash illusion), and health-related quality of life (SF-36). Over 12 weeks, the chiropractic group improved compared with the control group in choice stepping reaction time (119 milliseconds; 95% confidence interval [CI], 26-212 milliseconds; P = .01) and sound-induced flash illusion (13.5%; 95% CI, 2.9%-24.0%; P = .01). Ankle joint position sense improved across the 4- and 12-week assessments (0.20°; 95% CI, 0.01°-0.39°; P = .049). Improvements were also seen between weeks 4 and 12 in the SF-36 physical component of quality of life (2.4; 95% CI, 0.04-4.8; P = .04) compared with control. Sensorimotor function and multisensory integration associated with fall risk and the physical component of quality of life improved in older adults receiving chiropractic care compared with control. Future research is needed to investigate the mechanisms of action that contributed to the observed changes in this study and whether chiropractic care has an impact on actual falls risk in older adults.


Evolution by adapting surrogates.
To deal with complex optimization problems plagued with computationally expensive fitness functions, the use of surrogates to replace the original functions within the evolutionary framework is becoming a common practice. However, the appropriate datacentric approximation methodology to use for the construction of surrogate model would depend largely on the nature of the problem of interest, which varies from fitness landscape and state of the evolutionary search, to the characteristics of search algorithm used. This has given rise to the plethora of surrogate-assisted evolutionary frameworks proposed in the literature with ad hoc approximation/surrogate modeling methodologies considered. Since prior knowledge on the suitability of the data centric approximation methodology to use in surrogate-assisted evolutionary optimization is typically unavailable beforehand, this paper presents a novel evolutionary framework with the evolvability learning of surrogates (EvoLS) operating on multiple diverse approximation methodologies in the search. Further, in contrast to the common use of fitness prediction error as a criterion for the selection of surrogates, the concept of evolvability to indicate the productivity or suitability of an approximation methodology that brings about fitness improvement in the evolutionary search is introduced as the basis for adaptation. The backbone of the proposed EvoLS is a statistical learning scheme to determine the evolvability of each approximation methodology while the search progresses online. For each individual solution, the most productive approximation methodology is inferred, that is, the method with highest evolvability measure. Fitness improving surrogates are subsequently constructed for use within a trust-region enabled local search strategy, leading to the self-configuration of a surrogate-assisted memetic algorithm for solving computationally expensive problems. A numerical study of EvoLS on commonly used benchmark problems and a real-world computationally expensive aerodynamic car rear design problem highlights the efficacy of the proposed EvoLS in attaining reliable, high quality, and efficient performance under a limited computational budget.


Urocortin, a CRF-like peptide, restores key indicators of damage in the substantia nigra in a neuroinflammatory model of Parkinson's disease.
We have recently observed that the corticotrophin releasing hormone (CRF) related peptide urocortin (UCN) reverses key features of nigrostriatal damage in the hemiparkinsonian 6-hydroxydopamine lesioned rat. Here we have studied whether similar effects are also evident in the lipopolysaccaride (LPS) neuroinflammatory paradigm of Parkinson's disease (PD). To do this we have measured restoration of normal motor behaviour, retention of nigral dopamine (DA) and also tyrosine hydroxylase (TH) activity. Fourteen days following intranigral injections of LPS and UCN, rats showed only modest circling after DA receptor stimulation with apomorphine, in contrast to those given LPS and vehicle where circling was pronounced. In separate experiments, rats received UCN seven days following LPS, and here apomorphine challenge caused near identical circling intensity to those that received LPS and UCN concomitantly. In a similar and consistent manner with the preservation of motor function, UCN 'protected' the nigra from both DA depletion and loss of TH activity, indicating preservation of DA cells. The effects of UCN were antagonised by the non-selective CRF receptor antagonist alpha-helical CRF and were not replicated by the selective CRF2 ligand UCN III. This suggests that UCN is acting via CRF1 receptors, which have been shown to be anti-inflammatory in the periphery. Our data therefore indicate that UCN is capable of maintaining adequate nigrostriatal function in vivo, via CRF1 receptors following a neuro-inflammatory challenge. This has potential therapeutic implications in PD.


A rare case of limbic encephalitis with anti leucine-rich glioma inactivated-1 (LGI1) antibodies.
We report a case of limbic encephalitis with LGI1 antibodies and cranial MRI abnormality. A 41-year-old woman who presented with confusional state that had started 1 month ago. Brain magnetic resonance imaging revealed hyperintense signal in right basal ganglia followed by hyperintense signals in left hippocampus and bilateral basal ganglia half a month later. This case expands the spectrum of limbic encephalitis to include LGI1 antibodies and cranial MRI changing process.


The epidemiology of paediatric supracondylar fracture fixation: a population-based study.
The epidemiology of paediatric supracondylar fracture (SCF) fixation has not been evaluated at a population level. The purpose of this study was to: (1) determine the incidence density rate (IDR) of SCF fixation and (2) determine the rate of and risk factors for re-operation. Using administrative databases, all patients who underwent SCF fixation (closed reduction percutaneous pinning (CRPP) or open reduction (OR)) in Ontario between April 2002 and March 2010 were identified. Exclusion criteria included age (>12 years), a prior or concurrent non-SCF elbow fracture or previous humeral osteotomy. The overall IDR of SCF fixation and for subgroups of age, sex and season were calculated. A multivariate regression (immediate and short-term re-operation) and a Cox proportional hazards model (long-term re-operation) were used to identify patient, injury and provider factors that influenced re-operation risk and were reported as odds ratios or hazard ratios (HRs) with 95% confidence intervals (CIs), respectively. A total of 3235 patients with a median age of 6.0 years (interquartile range (IQR): 3.0) underwent SCF fixation. The median follow-up was 6.0 years (IQR: 3.7). The majority underwent a CRPP (78.7%) which were performed after hours (75.6%). The overall IDR was 20.7/100,000 person-years (py), but it varied significantly by season and age. Re-operation was uncommon in the immediate (1.0%), short-term (1.4%) and long-term (0.3%) follow-up period. As compared to CRPP, patients who underwent OR were more likely to undergo early nerve exploration (odds ratio: 7.8 (CI: 3.0-20.6)) and re-operation in the long term (HR: 3.0 (CI: 1.0-8.7)). Increased surgeon volume of SCF fixation was protective against repeat fixation (odds ratio: 0.9 (CI: 0.9-1.0)) and re-operation in the long term (HR: 0.9 (CI: 0.8-1.0)). While SCF fixation is common, the rate of re-operation is low. No differences existed between the sexes and a higher volume of fixations occurred during the summer months.


Results of stereotactic radiosurgery in patients with hormone-producing pituitary adenomas: factors associated with endocrine normalization.
The goal of this study was to determine factors associated with endocrine normalization after radiosurgery is performed in patients with hormone-producing pituitary adenomas. Between 1990 and 1999, 43 patients with hormone-producing pituitary adenomas underwent radiosurgery: 26 patients with growth hormone (GH)-producing tumors, nine with adrenocorticotrophic hormone-producing tumors, seven with tumors that produced prolactin (PRL) alone, and one with a tumor that secreted both GH and PRL. The median patient age was 42 years. Thirty-seven patients (86%) had undergone surgery earlier and in 30 (70%) there was tumor extension into the cavernous sinus. The product-limit method was used to calculate endocrine normalization while patients were not receiving any hormone-suppressive medication. The median follow-up period after radiosurgery was 36 months (range 12-108 months). In 20 patients (47%) there was normalization of hormone secretion at a median of 14 months (range 2-44 months) after radiosurgery; no correlation was found between tumor type and cure. Actuarial cure rates were 20, 32, and 61% at 1, 2, and 4 years posttreatment. Multivariate analysis demonstrated that the absence of hormone-suppressive medications at the time of radiosurgery (relative risk 8.9, 95% confidence interval [CI] 1.2-68.7, p = 0.04) and maximum radiation doses greater than 40 Gy (relative risk 3.9, 95% CI 1.3-11.7, p = 0.02) correlated with an endocrine cure. A new anterior pituitary deficiency developed in seven patients (16%), temporal lobe necrosis was identified in two patients, an asymptomatic internal carotid artery stenosis was detected in two patients, and unilateral blindness occurred in one patient. Radiosurgery provides an endocrine cure for many patients with persistent or recurrent hormone-producing pituitary adenomas. Further study is needed to determine whether pituitary hormone-suppressive medications have a radioprotective effect.


Cortical thickness and resting-state cardiac function across the lifespan: A cross-sectional pooled mega-analysis.
Understanding the association between autonomic nervous system [ANS] function and brain morphology across the lifespan provides important insights into neurovisceral mechanisms underlying health and disease. Resting-state ANS activity, indexed by measures of heart rate [HR] and its variability [HRV] has been associated with brain morphology, particularly cortical thickness [CT]. While findings have been mixed regarding the anatomical distribution and direction of the associations, these inconsistencies may be due to sex and age differences in HR/HRV and CT. Previous studies have been limited by small sample sizes, which impede the assessment of sex differences and aging effects on the association between ANS function and CT. To overcome these limitations, 20 groups worldwide contributed data collected under similar protocols of CT assessment and HR/HRV recording to be pooled in a mega-analysis (N = 1,218 (50.5% female), mean age 36.7 years (range: 12-87)). Findings suggest a decline in HRV as well as CT with increasing age. CT, particularly in the orbitofrontal cortex, explained additional variance in HRV, beyond the effects of aging. This pattern of results may suggest that the decline in HRV with increasing age is related to a decline in orbitofrontal CT. These effects were independent of sex and specific to HRV; with no significant association between CT and HR. Greater CT across the adult lifespan may be vital for the maintenance of healthy cardiac regulation via the ANS-or greater cardiac vagal activity as indirectly reflected in HRV may slow brain atrophy. Findings reveal an important association between CT and cardiac parasympathetic activity with implications for healthy aging and longevity that should be studied further in longitudinal research.


Niacin induces PPARgamma expression and transcriptional activation in macrophages via HM74 and HM74a-mediated induction of prostaglandin synthesis pathways.
HM74 and HM74a have been identified as receptors for niacin. HM74a mediates the pharmacological anti-lipolytic effects of niacin in adipocytes by reducing intracellular cyclic AMP (cAMP) and inhibiting release of free fatty acids into the circulation. In macrophages, niacin induces peroxisome proliferator-activated receptor gamma (PPARgamma)-dependent and cAMP-dependent expression of genes mediating reverse cholesterol transport, although via an unidentified receptor. We describe constitutive expression of HM74a mRNA and hypoxia- and IFNgamma-inducible expression of HM74 and HM74a in human monocytic cell lines and primary cells in culture. In U937 cells niacin-induced expression of 15-deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)), the most potent endogenous ligand of PPARgamma. Both niacin and the structurally distinct HM74/HM74a ligand acifran-induced nuclear expression of PPARgamma protein and enhanced PPARgamma transcriptional activity. Niacin-induced PPARgamma transcriptional activity was pertussis toxin sensitive and required activity of phospholipase A(2) (EC 18.104.22.168), cyclo-oxygenase (EC 22.214.171.124) and prostaglandin D(2) synthase (EC 188.8.131.52). Niacin also induced PPARgamma transcriptional activity in HM74 and HM74a CHO cell transfectants, although not in vector-only control cells. This was sensitive to pertussis toxin and to inhibition of phoshoplipase A(2) and cyclo-oxygenase activity. Additionally, niacin increased intracellular cAMP in U937 via a pertussis toxin and cyclo-oxygenase-sensitive mechanism. These results indicate that HM74 and HM74a can mediate macrophage responses to niacin via activation of the prostaglandin synthesis pathway and induction and activation of PPARgamma. This suggests a novel mechanism(s) mediating the clinical effects of pharmacological doses of niacin.


Acting, seeing, and conscious awareness.
We argue that there is a relation between the judgements that 'I did it' and 'I saw it'. Both are statements are about the individual, not just the world. We show that the dorsal prefrontal cortex is activated both when human subjects judge that they are the agents of their actions and when they judge that they are confident that they have seen a masked visual stimulus. Macaque monkeys have also been taught to report whether they have or have not seen visual stimuli and cells can be found in the dorsal prefrontal cortex that distinguish between 'seen' and 'not seen'. The judgement is abstract in that it applies largely irrespective of the nature and location of the stimulus. We suggest that the reason why the prefrontal cortex is involved is that it evolved in primates, adapted by searching for fruit and leaves and using their hands to retrieve them. There is cell activity in the dorsal prefrontal cortex that relates to eye movements, covert attention and visual search; activity that relates to learning abstract rules; and activity that relates to the planning of the hand movements that are appropriate. We propose that this is the reason why this area is involved in making judgements about both agency and visual detection.


Population trends in Pacific Oceanic sharks and the utility of regulations on shark finning.
Accurate assessment of shark population status is essential for conservation but is often constrained by limited and unreliable data. To provide a basis for improved management of shark resources, we analyzed a long-term record of species-specific catches, sizes, and sexes of sharks collected by onboard observers in the western and central Pacific Ocean from 1995 to 2010. Using generalized linear models, we estimated population-status indicators on the basis of catch rate and biological indicators of fishing pressure on the basis of median size to identify trends for blue (Prionace glauca), mako (Isurus spp.), oceanic whitetip (Carcharhinus longimanus), and silky (Carcharhinus falciformis) sharks. Standardized catch rates of longline fleets declined significantly for blue sharks in the North Pacific (by 5% per year [CI 2% to 8%]), for mako sharks in the North Pacific (by 7% per year [CI 3% to 11%]), and for oceanic whitetip sharks in tropical waters (by 17% per year [CI 14% to 20%]). Median lengths of silky and oceanic whitetip sharks decreased significantly in their core habitat, and almost all sampled silky sharks were immature. Our results are consistent with results of analyses of similar data sets. Combined, these results and evidence of targeted fishing for sharks in some regional fisheries heighten concerns for sustainable utilization, particularly for oceanic whitetip and North Pacific blue sharks. Regional regulations that prohibit shark finning (removal of fins and discarding of the carcass) were enacted in 2007 and are in many cases the only form of control on shark catches. However, there is little evidence of a reduction of finning in longline fisheries. In addition, silky and oceanic whitetip sharks are more frequently retained than finned, which suggests that even full implementation of and adherence to a finning prohibition may not substantially reduce mortality rates for these species. We argue that finning prohibitions divert attention from assessing whether catch levels are sustainable and that the need for management of sharks should not be addressed by measures that are simple to implement but complex to enforce and evaluate.


Genomic and transcriptomic heterogeneity in metaplastic carcinomas of the breast.
Metaplastic breast cancer (MBC) is a rare special histologic type of triple-negative breast cancer, characterized by the presence of neoplastic cells showing differentiation towards squamous epithelium and/or mesenchymal elements. Here we sought to define whether histologically distinct subgroups of MBCs would be underpinned by distinct genomic and/or transcriptomic alterations. Microarray-based copy number profiling identified limited but significant differences between the distinct MBC subtypes studied here, despite the limited sample size (n = 17). In particular, we found that, compared to MBCs with chondroid or squamous cell metaplasia, MBCs with spindle cell differentiation less frequently harbored gain of 7q11.22-23 encompassing CLDN3 and CLDN4, consistent with their lower expression of claudins and their association with the claudin-low molecular classification. Microarray-based and RNA-sequencing-based gene expression profiling revealed that MBCs with spindle cell differentiation differ from MBCs with chondroid or squamous cell metaplasia on the expression of epithelial-to-mesenchymal transition-related genes, including down-regulation of CDH1 and EPCAM. In addition, RNA-sequencing revealed that the histologic patterns observed in MBCs are unlikely to be underpinned by a highly recurrent expressed fusion gene or a pathognomonic expressed mutation in cancer genes. Loss of PTEN expression or mutations affecting PIK3CA or TSC2 observed in 8/17 MBCs support the contention that PI3K pathway activation plays a role in the development of MBCs. Our data demonstrate that despite harboring largely similar patterns of gene copy number alterations, MBCs with spindle cell, chondroid and squamous differentiation are distinct at the transcriptomic level but are unlikely to be defined by specific pathognomonic genetic alterations.


Cutaneous vasodilation during dorsal column stimulation is mediated by dorsal roots and CGRP.
Dorsal column stimulation (DCS) is used clinically to provide pain relief from peripheral vascular disease and has the benefit of increasing cutaneous blood flow to the affected lower extremities. The purpose of this study was to examine the role of dorsal roots, calcitonin gene-related peptide (CGRP), and substance P in the cutaneous vasodilation induced by DCS. Male rats were anesthetized with pentobarbital sodium (60 mg/kg ip). A unipolar ball electrode was placed unilaterally on the spinal cord at the L1-L2 spinal segment. Blood flow was recorded in each hindpaw foot pad with laser Doppler flowmeters. Blood flow responses were assessed during 1 min of DCS (either 0.2 mA subdural or 0.6 mA epidural at 50 Hz, 0.2-ms pulse duration). Dorsal rhizotomy of L3-L5 (n = 5) abolished the cutaneous vasodilation to subdural DCS, whereas removal of T10-T12 (n = 5) and T13-L2 dorsal roots (n = 5) did not attenuate the DCS-induced vasodilation. The CGRP antagonist, CGRP-(8-37) (2.6 mg/kg iv, n = 7), eliminated the epidural DCS-induced vasodilation, whereas the substance P receptor antagonist, CP-96345 (1 mg/kg iv, n = 6), had no effect. In summary, L3-L5 dorsal roots and CGRP are essential for the DCS-induced vasodilation. We propose that DCS antidromically activates afferent fibers in the dorsal roots, thus causing peripheral release of CGRP, which produces cutaneous vasodilation.


A major quantitative trait locus for rice yellow mottle virus resistance maps to a cluster of blast resistance genes on chromosome 12.
ABSTRACT Two doubled-haploid rice populations, IR64/Azucena and IRAT177/ Apura, were used to identify markers linked to rice yellow mottle virus (RYMV) resistance using core restriction fragment length polymorphism (RFLP) maps. Resistance was measured by mechanical inoculation of 19-day-old seedlings followed by assessment of virus content by enzyme-linked immunosorbent assay tests 15 days after inoculation. IR64/Azucena and IRAT177/Apura populations, 72 and 43 lines, respectively, were evaluated, and resistance was found to be polygenic. Resistance was expressed as a slower virus multiplication, low symptom expression, and limited yield loss when assessed at the field level. Bulked segregant analysis using the IR64/Azucena population identified a single random amplified polymorphic DNA marker that mapped on chromosome 12 and corresponded to a major quantitative trait locus (QTL) evidenced by interval mapping. When pooling RFLP data, integrated mapping of this chromosome revealed that the QTL was common to the two populations and corresponded to a small chromosomal segment known to contain a cluster of major blast resistance genes. This region of the genome also reflected the differentiation observed at the RFLP level between the subspecies indica and japonica of Oryza sativa. This is consistent with the observation that most sources of RYMV resistance used in rice breeding are found in upland rice varieties that typically belong to the japonica subspecies.


Tissue artifact removal from respiratory signals based on empirical mode decomposition.
On-line measurement of respiration plays an important role in monitoring human physical activities. Such measurement commonly employs sensing belts secured around the rib cage and abdomen of the test object. Affected by the movement of body tissues, respiratory signals typically have a low signal-to-noise ratio. Removing tissue artifacts therefore is critical to ensuring effective respiration analysis. This paper presents a signal decomposition technique for tissue artifact removal from respiratory signals, based on the empirical mode decomposition (EMD). An algorithm based on the mutual information and power criteria was devised to automatically select appropriate intrinsic mode functions for tissue artifact removal and respiratory signal reconstruction. Performance of the EMD-algorithm was evaluated through simulations and real-life experiments (N = 105). Comparison with low-pass filtering that has been conventionally applied confirmed the effectiveness of the technique in tissue artifacts removal.


How social-class stereotypes maintain inequality.
Social class stereotypes support inequality through various routes: ambivalent content, early appearance in children, achievement consequences, institutionalization in education, appearance in cross-class social encounters, and prevalence in the most unequal societies. Class-stereotype content is ambivalent, describing lower-SES people both negatively (less competent, less human, more objectified), and sometimes positively, perhaps warmer than upper-SES people. Children acquire the wealth aspects of class stereotypes early, which become more nuanced with development. In school, class stereotypes advantage higher-SES students, and educational contexts institutionalize social-class distinctions. Beyond school, well-intentioned face-to-face encounters ironically draw on stereotypes to reinforce the alleged competence of higher-status people and sometimes the alleged warmth of lower-status people. Countries with more inequality show more of these ambivalent stereotypes of both lower-SES and higher-SES people. At a variety of levels and life stages, social-class stereotypes reinforce inequality, but constructive contact can undermine them; future efforts need to address high-status privilege and to query more heterogeneous samples.


Vegetable-derived bioactive nitrate and cardiovascular health.
Vegetable derived nitrate is now recognised as an important bioactive phytochemical with cardioprotective properties. Nitrate, through the recently described enterosalivary nitrate-nitrite-nitric oxide (NO) pathway, increases NO, a molecule pivotal for cardiovascular health. Clinical trials have observed that dietary nitrate has similar effects to NO when supplied exogenously. These effects include reduced blood pressure and improvements in other markers of vascular health such as endothelial function, arterial stiffness, ischemia reperfusion injury, blood flow, and platelet aggregation. Few observational studies, however, have examined dietary nitrate intake and long term cardiovascular health outcomes. This represents a significant gap in the literature. There is also a lingering concern about a possible carcinogenic effect of nitrate intake. Additionally, a number of potential factors that could impact nitrate to nitrite to NO reduction have been identified. This review will provide an overview of the evidence to date that nitrate, through its effects on endogenous NO and vascular health, is an important bioactive cardioprotective component of a diet rich in vegetables.


Viral Injection and Cranial Window Implantation for In Vivo Two-Photon Imaging.
Two-photon (2P) imaging has proven to be a powerful tool for investigating neural structure and function both in brain slices and in intact systems. In vivo 2P imaging presents significant challenges in sample preparation, which are exacerbated in non-murine species. Here, we describe procedures for the effective virally mediated labeling of neurons and for the implantation of cranial windows for imaging. The procedures described here are applicable to a range of species, including mice, and are routinely used in ferrets and tree shrews to provide large-scale labeling of cortical volumes and high-quality imaging data.


Spatial correlations between the vacuolation, prion protein deposits, and surviving neurons in the cerebral cortex in sporadic Creutzfeldt-Jakob disease.
In the cerebral cortex of cases of sporadic Creutzfeldt-Jakob disease (sCJD), the vacuolation (spongiform change) and PrP deposits are aggregated into clusters which are regularly distributed parallel to the pia mater. The objective of the present study was to determine the spatial relationships between the clusters of the vacuoles and PrP deposits and between the pathological changes and variations in the density of surviving neurons. In areas with low densities of pathological change, clusters of vacuoles were spatially correlated with the surviving neurons and not with the PrP deposits. By contrast, in more significantly affected areas, clusters of vacuoles were spatially correlated with those of the PrP deposits and not with the surviving neurons. In addition, areas with a high density of vacuoles and a low density of PrP deposits exhibited no spatial correlations between the variables. These data suggest that the spatial relationships between the vacuolation, PrP deposits and surviving neurons in sCJD depend on the density of lesions present. Differences in the pattern of correlation may reflect the developmental stage of the pathology in particular cortical areas.


Evaluation and mechanism analysis of HIV prevention programme using resilience framework among female sex workers: A randomised controlled trial.
Evidence shows traditional sexual harm reduction for female sex workers (FSW) based on health behaviour theories is effective but short-lived. This study aimed to evaluate and understand the effectiveness of a resilience-promoting programme in improving psychological health and, ultimately, safe sex practice. A randomised controlled trial was conducted at three Hong Kong-based non-governmental organisations. 127 sex workers were recruited and randomly assigned to the intervention or control groups. The former received a six-session resilience-promoting programme designed to improve self-esteem, self-efficacy and coping skills, whereas the latter had the usual care. Between-group differences in psychological outcomes and condom use were tested using the intention-to-treat, with ANOVA and chi-square tests, measured at baseline, post-intervention and 3-month follow-ups. Multiple mediation analysis was used to examine how the intervention worked through resilience factors. Significant between-group improvements in adaptive coping (F 1,119 = 5.82, p < .05) and reduction in psychological distress (F 1,118 = 5.00, p < .05) were seen at post-intervention and 3-month follow-ups, with significant time × group interaction changes suggesting the changes occurred at different rates between the two groups. Condom use during the last transactions had increased in the intervention group and the rate of consistent condom use during transactions improved in the intervention group at follow-ups (χ 2 = 4.35, p < .05). Self-esteem and resilience significantly mediated the effect of intervention at reducing psychological distress. These findings suggest that resilience improves the psychological health and general wellbeing of Chinese FSWs.


POL and PLL1 phosphatases are CLAVATA1 signaling intermediates required for Arabidopsis shoot and floral stem cells.
The post-embryonic development of above-ground tissues in plants is dependent upon the maintenance and differentiation of stem cells at the shoot meristem. The Arabidopsis WUSCHEL (WUS) transcription factor establishes an organizing center within the shoot meristem that is essential for specification of stem-cell identity in overlying cells. The CLAVATA (CLV) signaling pathway, including the CLV1 receptor-kinase, promotes the differentiation of stem cells by limiting the WUS expression domain, yet the mechanism of CLV signaling is largely unknown. Previously, we have shown that mutations in two protein phosphatases, POLTERGEIST (POL) and PLL1, partially suppress clv mutant phenotypes. Here, we demonstrate that POL and PLL1 are integral components of the CLV1 signaling pathway. POL and PLL1 are essential for stem-cell specification, and can also block stem-cell differentiation when overexpressed. We provide extensive evidence that POL and PLL1 act downstream of CLV signaling to maintain WUS expression and that they regulate WUS at a transcriptional level. Our findings suggest that POL and PLL1 are central players in regulating the balance between stem-cell maintenance and differentiation, and are the closest known factors to WUS regulation in the shoot meristem.


Family and peer influences on adjustment among Chinese, Filipino, and White youth.
Little is known about the influence of parent-adolescent relationships and peer behavior on emotional distress and risky behaviors among Asian American adolescents; in particular, cross-cultural and longitudinal examinations are missing from the extant research. To test and compare a theoretical model examining the influence of family and peer factors on adolescent distress and risky behavior over time, using a nationally representative sample of Chinese, Filipino, and White adolescents. Data were utilized from Waves I (1994) and II (1995) of the National Longitudinal Study on Adolescent Health; the sample is composed of 194 Chinese, 345 Filipino, and 395 White adolescents and weighted to correct for design effects, yielding a nationally representative sample. Structural equation modeling was used to test the theoretical model for each ethnic group separately followed by multiple-group analyses. The measurement model was examined for each ethnic group, using both unweighted and weighted samples, and was deemed equivalent across groups. Tests of the theoretical model by ethnicity revealed that for each group, family bonds have significant negative effects on emotional distress and risky behaviors. For Filipino and White youth, peer risky behaviors influenced risky behaviors. Multiple-group analyses of the theoretical model indicated that the three ethnic groups did not differ significantly from one another. Findings suggest that family bonds and peer behavior exert significant influences on psychological and behavioral outcomes in Asian American youth and that these influences appear to be similar with White adolescents. Future research should be directed toward incorporating variables known to contribute to the impact of distress and risky behaviors in model testing and validating findings from this study.


Nano in cancer: linking chemistry, biology, and clinical applications in vivo.
Development of nanoparticle agents for cancer therapeutics and diagnostics is steadily progressing and was the subject of the inaugural conference entitled, "Nano in Cancer," held during January 12-15, 2011, in Miami, FL. The meeting program was developed by co-chairs David Piwnica-Worms (Washington University in St. Louis), Jan Schnitzer (Proteogenomics Research Institute for Systems Medicine, San Diego), and Karen Wooley (Texas A&M University). Topics discussed for nanoparticle platforms under development included: nanotechnologies for cancer diagnostics and imaging, overcoming in vivo barriers, therapeutic nanoparticles and clinical prospects, and safety issues for nanotechnologies. Two important concepts emerged from this meeting. The first was the urgent need for uniform standards and protocols for nanoparticle characterization in vitro and in vivo, and the second was the continued need for discovery of definitive targets for tumor-directed nanoparticles across various cancers.


Incidence and nature of dysphagia in polio survivors.
Questionnaires pertaining to swallowing function were mailed to 220 members of postpolio support groups in Connecticut. Of the 109 responses, 80 individuals reported having no difficulty with swallowing, while 29 reported having either intermittent or consistent swallowing problems. Twenty-one of the 29 were seen for videofluoroscopic swallowing studies and pulmonary function testing. The swallowing studies showed that 43% of these individuals had difficulty with bolus control, 19% with delayed swallow response, and 81% with decreased pharyngeal transit. Although none of these individuals were observed to aspirate, two were judged to be at significant risk. Incidence of dysphagia within the group of polio survivors was estimated to be approximately 18%. Seventeen of the 20 postpolio subjects with dysphagia also demonstrated decreased breathing capacity. Although moderately to severely depressed values in the pulmonary function measures accompanied moderate dysphagia in certain postpolio individuals, reduced values in these same measures were also present in individuals with minimal swallowing dysfunction. Therefore, although impaired breathing may complicate swallowing dysfunction and vice versa, it does not appear that one can be predicated from the other. Management of dysphagia in postpolio individuals is discussed.


Pragmatic randomized trial of antenatal intervention to prevent post-natal depression by reducing psychosocial risk factors.
Social support theory and observational risk factor studies suggest that increased antenatal psychosocial support could prevent post-natal depression. We used empirical knowledge of risk and protective factors for post-natal depression not employed previously in order to develop and evaluate an antenatal preventive intervention. We conducted a pragmatic randomized controlled trial in antenatal clinics. We screened 1300 primiparous women and 400 screened positive, 69 screen-positive women were untraceable or not eligible. Of 292 women who completed baseline assessment, 209 consented to randomization, of these 190 provided outcome data 3 months post-natally. 'Preparing for Parenthood', a structured antenatal risk factor reducing intervention designed to increase social support and problem-solving skills, was compared with routine antenatal care only. We compared the percentage depressed at 3 months after childbirth using the self-completion General Health Questionnaire Depression scale and Edinburgh Post-natal Depression Scale (EPDS), and the Schedules for Clinical Assessment in Neuropsychiatry a systematic clinical interview. Assignment to the intervention group did not significantly impact on post-natal depression (odds ratio for GHQ-Depression 1.22 (95% CI 0.63-2.39), P = 0.55) or on risk factors for depression. Forty-five per cent of the intervention group women attended sufficient sessions to be likely to benefit from intervention if effective. Attenders benefited no more than non-attenders. Prevention services targeting post-natal depression should not implement antenatal support programmes on these lines until further research has demonstrated the feasibility and effectiveness of such methods. The development of novel, low cost interventions effective in reducing risk factors should be completed before further trial evaluation.


Vasoactivity of the major intracranial arteries in newborn infants.
Blood flow to the head and arms was measured by a new volumetric Doppler technique in 30 preterm infants and 10 full term infants. At least 80% of this blood flow is considered to perfuse the brain. At the same time mean blood velocity in the middle and anterior cerebral arteries was measured by Duplex Doppler scanning. While blood flow to the head and arms remained substantially constant in both groups over the first 48 hours of life (60 ml/100 g brain weight/min), blood velocity doubled in both cerebral arteries in the preterm group. The term infants had higher blood velocities in both arteries at all times, but the velocities also increased over 48 hours, although to a lesser extent than in the preterm group. These findings imply that the major intracranial arteries are themselves vasoactive, being dilated at birth and subsequently constricting, possibly as an autoregulatory response to rising arterial blood pressure. While vasodilated, the cerebral arteries will be less efficient at damping pressure transients, placing preterm infants at particular risk of periventricular haemorrhage during the early hours of life. The implications for possible approaches to the prevention of cerebral haemorrhage are discussed.


Accuracy of CT in the diagnosis of pulmonary embolism: a systematic literature review.
We sought to summarize systematically the published evidence describing the accuracy of contrast-enhanced helical CT for diagnosing pulmonary embolism. We selected all systematic reviews published before December 2003 that evaluated the accuracy of CT angiography for the diagnosis of pulmonary embolism. We also selected all prospective studies from the same time period in the primary literature in which all subjects underwent both CT and conventional angiography, the latter being considered the reference standard. Articles were identified through a computerized MEDLINE search and by other means. The quality and content of each article were evaluated independently by pairs of researchers. Six systematic reviews and eight primary studies were selected. The combined sensitivities of CT for detecting pulmonary embolism ranged from 66% to 93% across the systematic reviews and the combined specificities ranged from 89% to 97%. Only one of the reviews reported a combined sensitivity of greater than 90%. Among the eight primary studies, the sensitivities ranged from 45% to 100% and specificities ranged from 78% to 100%. Only three of the eight primary studies reported a sensitivity greater than 90%. None of the primary studies used scanners with four or more detectors. A systematic literature review revealed a wide range of reported sensitivities, only a minority of which exceeded 90%. Pooled estimates of sensitivity and specificity reported by systematic literature reviews should be interpreted with caution because of potential selection bias and heterogeneity in the reviewed studies. Accuracy studies of recent generations of MDCT scanners are not yet available despite the current dissemination of this technology.


Respiratory drive during carbachol challenge in allergic sheep.
Diaphragmatic activity is the standard assessment of respiratory neural output but is difficult to measure and cannot be used for long-term clinical monitoring. The tidal breathing minute ventilation (V') and mean inspiratory flow (VT/tI) reflect respiratory drive and can be monitored non-invasively using respiratory inductive plethysmography (RIP). Recent findings indicate that peak inspiratory acceleration (PIA) correlates to diaphragmatic activity during CO2 rebreathing in piglets. The aim of the present study was to assess whether tidal breathing peak inspiratory acceleration reflects respiratory drive during carbachol challenge. Aerosolized carbachol was administered to 15 allergic sheep until pulmonary resistance increased by at least 400%. After threshold dose, slight increases in V' and VT/tI were seen (8 and 5%, respectively; P=NS), while PIA increased by 30% (P<0.05). The change in PIA was closely correlated to changes in V'[bar over] and VT/tI (r=0.73, P<0.01 and r=0.88, P<0.001, respectively). In conclusion, peak inspiratory acceleration reflected respiratory drive during bronchoprovocation. Further, this new measure of drive has the potential to accurately estimate drive in chronic obstructive lung disease where intrinsic positive end expiratory pressure prevents accurate selection of beginning inspiration necessary for computation of, e.g. VT/tI.


Drought reduces floral resources for pollinators.
Climate change is predicted to result in increased occurrence and intensity of drought in many regions worldwide. By increasing plant physiological stress, drought is likely to affect the floral resources (flowers, nectar and pollen) that are available to pollinators. However, little is known about impacts of drought at the community level, nor whether plant community functional composition influences these impacts. To address these knowledge gaps, we investigated the impacts of drought on floral resources in calcareous grassland. Drought was simulated using rain shelters and the impacts were explored at multiple scales and on four different experimental plant communities varying in functional trait composition. First, we investigated the effects of drought on nectar production of three common wildflower species (Lathyrus pratensis, Onobrychis viciifolia and Prunella vulgaris). In the drought treatment, L. pratensis and P. vulgaris had a lower proportion of flowers containing nectar and O. viciifolia had fewer flowers per raceme. Second, we measured the effects of drought on the diversity and abundance of floral resources across plant communities. Drought reduced the abundance of floral units for all plant communities, irrespective of functional composition, and reduced floral species richness for two of the communities. Functional diversity did not confer greater resistance to drought in terms of maintaining floral resources, probably because the effects of drought were ubiquitous across component plant communities. The findings indicate that drought has a substantial impact on the availability of floral resources in calcareous grassland, which will have consequences for pollinator behaviour and populations.


Implications of Early Decline in eGFR due to Intensive BP Control for Cardiovascular Outcomes in SPRINT.
The Systolic BP Intervention Trial (SPRINT) found that intensive versus standard systolic BP control (targeting <120 or <140 mm Hg, respectively) reduced the risks of death and major cardiovascular events in persons with elevated cardiovascular disease risk. However, the intensive intervention was associated with an early decline in eGFR, and the clinical implications of this early decline are unclear. In a post hoc analysis of SPRINT, we defined change in eGFR as the percentage change in eGFR at 6 months compared with baseline. We performed causal mediation analyses to separate the overall effects of the randomized systolic BP intervention on the SPRINT primary cardiovascular composite and all-cause mortality into indirect effects (mediated by percentage change in eGFR) and direct effects (mediated through pathways other than percentage change in eGFR). About 10.3% of the 4270 participants in the intensive group had a ≥20% eGFR decline versus 4.4% of the 4256 participants in the standard arm (P<0.001). After the 6-month visit, there were 591 cardiovascular composite events during 27,849 person-years of follow-up. The hazard ratios for total effect, direct effect, and indirect effect of the intervention on the cardiovascular composite were 0.67 (95% confidence interval [95% CI], 0.56 to 0.78), 0.68 (95% CI, 0.57 to 0.79), and 0.99 (95% CI, 0.95 to 1.03), respectively. All-cause mortality results were similar. Although intensive systolic BP lowering resulted in greater early decline in eGFR, there was no evidence that the reduction in eGFR owing to intensive systolic BP lowering attenuated the beneficial effects of this intervention on cardiovascular events or all-cause mortality.


Dissolution rate enhancement, in vitro evaluation and investigation of drug release kinetics of chloramphenicol and sulphamethoxazole solid dispersions.
Formulation of solid dispersions is one of the effective methods to increase the rate of solubilization and dissolution of poorly soluble drugs. Solid dispersions of chloramphenicol (CP) and sulphamethoxazole (SX) as model drugs were prepared by melt fusion method using polyethylene glycol 8000 (PEG 8000) as an inert carrier. The dissolution rate of CP and SX were rapid from solid dispersions with low drug and high polymer content. Characterization was performed using fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC) and scanning electron microscopy (SEM). FTIR analysis for the solid dispersions of CP and SX showed that there was no interaction between PEG 8000 and the drugs. Hyper-DSC studies revealed that CP and SX were converted into an amorphous form when formulated as solid dispersion in PEG 8000. Mathematical analysis of the release kinetics demonstrated that drug release from the various formulations followed different mechanisms. Permeability studies demonstrated that both CP and SX when formulated as solid dispersions showed enhanced permeability across Caco-2 cells and CP can be classified as well-absorbed compound when formulated as solid dispersions.


Differential glucose uptake in quadriceps and other leg muscles during one-legged dynamic submaximal knee-extension exercise.
One-legged dynamic knee-extension exercise (DKE) is a widely used model to study the local cardiovascular and metabolic responses to exercise of the quadriceps muscles. In this study, we explored the extent to which different muscles of the quadriceps are activated during exercise using positron emission tomography (PET) determined uptake of [(18)F]-fluoro-deoxy-glucose (GU) during DKE. Five healthy male subjects performed DKE at 25 W for 35 min and both the contracting and contralateral resting leg were scanned with PET from mid-thigh and distally. On average, exercise GU was the highest in the vastus intermedius (VI) and lowest in the vastus lateralis (VL; VI vs VL, p < 0.05), whereas the coefficient of variation was highest in VL (VL vs VI, p < 0.05). Coefficient of variation between the mean values of the four quadriceps femoris (QF) muscles in the exercising leg was 35 ± 9%. Compared to mean GU in QF (=100%), GU was on average 73% in VL, 84% in rectus femoris, 115% in vastus medialis, and 142% in VI. Variable activation of hamstring muscles and muscles of the lower leg was also observed. These results show that GU of different muscles of quadriceps muscle group as well as between individuals vary greatly during DKE, and suggests that muscle activity is not equal between quadriceps muscles in this exercise model. Furthermore, posterior thigh muscles and lower leg muscles are more active than hitherto thought even during this moderate exercise intensity.


SANTAVAC ™: A Novel Universal Antigen Composition for Developing Cancer Vaccines.
Development of a universal cancer vaccine for the prevention of all cancers has been under development for many years. Antiangiogenic cancer vaccines elicit immune responses with the potential of destroying tumor vasculature endothelial cells without affecting vasculature integrity in normal tissues. The methods used in the development of antigen compositions comprising these vaccines have been recently improved and described in this report in the context of SANTAVAC ™ development - the first cancer vaccine based on endothelial cell heterogeneity. The present report summarizes data related to SANTAVAC™ development, including technical key points associated with optimal SANTAVAC™ production, a description of the composition required for preparing cancer vaccines with the highest predicted efficacy and safety, and a strategy for SANTAVAC™ large-scale implementation. Patents related to SANTAVAC™ and other universal cancer vaccines are also described. SANTAVAC ™ was shown to be the most promising antigen composition for anti-cancer vaccination, allowing for immune targeting of the tumor vasculature in experimental models with a high predicted efficacy (up to 60), where efficacy represents the fold decrease in the number of endothelial cells with a tumor-induced phenotype and directly related to predicted arrest of tumor growth. The use of SANTAVAC ™ as a universal antigenic composition may spur vaccine development activities resulting in a set of therapeutic or prophylactic vaccines against different types of solid cancers.


Distinctive patterns of microRNA expression in primary muscular disorders.
The primary muscle disorders are a diverse group of diseases caused by various defective structural proteins, abnormal signaling molecules, enzymes and proteins involved in posttranslational modifications, and other mechanisms. Although there is increasing clarification of the primary aberrant cellular processes responsible for these conditions, the decisive factors involved in the secondary pathogenic cascades are still mainly obscure. Given the emerging roles of microRNAs (miRNAs) in modulation of cellular phenotypes, we searched for miRNAs regulated during the degenerative process of muscle to gain insight into the specific regulation of genes that are disrupted in pathological muscle conditions. We describe 185 miRNAs that are up- or down-regulated in 10 major muscular disorders in humans [Duchenne muscular dystrophy (DMD), Becker muscular dystrophy, facioscapulohumeral muscular dystrophy, limb-girdle muscular dystrophies types 2A and 2B, Miyoshi myopathy, nemaline myopathy, polymyositis, dermatomyositis, and inclusion body myositis]. Although five miRNAs were found to be consistently regulated in almost all samples analyzed, pointing to possible involvement of a common regulatory mechanism, others were dysregulated only in one disease and not at all in the other disorders. Functional correlation between the predicted targets of these miRNAs and mRNA expression demonstrated tight posttranscriptional regulation at the mRNA level in DMD and Miyoshi myopathy. Together with direct mRNA-miRNA predicted interactions demonstrated in DMD, some of which are involved in known secondary response functions and others that are involved in muscle regeneration, these findings suggest an important role of miRNAs in specific physiological pathways underlying the disease pathology.


Keeping Up With the Orthopaedic In-Training Examination: National Survey on Orthopaedic Residency Training in Practice Management.
The evolving health care environment warrants its physicians to be competent in basic practice management (PM) areas. A manifestation of this importance was reflected in the inaugural inclusion of a PM subsection on the 2017 Orthopaedic In-Training Examination. The purpose of this orthopaedic resident national survey study was to gain insight on the current state of formal residency education in PM. This study surveyed 500 orthopaedic residents nationwide in 2016. Resident participation was online, anonymous, and voluntary. Only complete survey responses were included, yielding a 49.2% (246/500) response rate. The majority of orthopaedic residents (72.4%, 178/246) reported no formal education in PM topics, and 86.2% (212/246) responded that they do not receive direct feedback on individual accuracy of Current Procedural Terminology (CPT) code case logging. Of the residents without formal education in PM, 87.1% (155/178) desire its implementation. The evolving health care system is becoming increasingly reliant on physicians to provide cost-effective, value-based health care to its patients. Consideration should be given to formally incorporating basic teaching elements on important PM topics at the residency training level. (Journal of Surgical Orthopaedic Advances 28(2):81-88, 2019).


Advances in radical probe mass spectrometry for protein footprinting in chemical biology applications.
Radical Probe Mass Spectrometry (RP-MS), first introduced in 1999, utilizes hydroxyl radicals generated directly within aqueous solutions using synchrotron radiolysis, electrical discharge, and photochemical laser sources to probe protein structures and their interactions. It achieves this on millisecond and submillisecond timescales that can be used to capture protein dynamics and folding events. Hydroxyl radicals are ideal probes of solvent accessibility as their size approximates a water molecule. Their high reactivity results in oxidation at a multitude of amino acid side chains providing greater structural information than a chemical cross-linker that reacts with only one or few residues. The oxidation of amino acid side chains occurs at rates in accord with the solvent accessibility of the residue so that the extent of oxidation can be quantified to reveal a three-dimensional map or footprint of the protein's surface. Mass spectrometry is central to this analysis of chemical oxidative labelling. This tutorial review, some 15 years on from the first reports, highlights the development and significant growth of the application of RP-MS including its validation and utility with ion-mobility mass spectrometry (IM-MS), the use of RP-MS data to help model protein complexes, studies of the onset of oxidative damage, and more recent advances that enable high throughput applications through simultaneous protein oxidation and on-plate deposition. The accessibility of the RP-MS technology, by means of a modified electrospray ionization source, enables the approach to be implemented in many laboratories to address a wide range of applications in chemical biology.


Effects of Exercise Training in Hypoxia Versus Normoxia on Vascular Health.
Exercise training (ExT) prompts multiple beneficial adaptations associated with vascular health, such as increases in skeletal muscle capillarization and vascular dilator function and decreases in arterial stiffness. However, whether ExT performed in hypoxic conditions induces enhanced effects is unclear. We sought to systematically review the literature and determine whether hypoxic ExT leads to superior vascular adaptations compared with normoxic ExT. We searched MEDLINE, Scopus, and Web of Science from their inception until September 2015 for articles assessing vascular adaptations to ExT performed under hypoxic and normoxic conditions. We performed meta-analyses to determine the standardized mean difference (SMD) between the effects of ExT performed in hypoxia versus normoxia on vascular adaptations. We assessed heterogeneity among studies using I 2 statistics and evaluated publication bias via the Begg and Mazumdar's rank correlation test and Egger's regression test. After systematic review, we included 21 controlled studies, including a total of 331 individuals (mean age 19-57 years, 265 males). ExT programs primarily consisted of cycling endurance training performed in normobaric hypoxia or normoxia; duration ranged from 3 to 10 weeks. The exercise intensity was similar in relative terms in the groups trained in hypoxia and normoxia in the majority of studies (17 of 21). After data pooling, skeletal muscle capillarization (n = 182, SMD = 0.40, 95 % confidence interval [CI] 0.10-0.70; P = 0.01) and vascular dilator function (n = 71, SMD = 0.67, 95 % CI 0.17-1.18; P = 0.009) but not arterial stiffness (n = 112, SMD = -0.03, 95 % CI -0.69 to 0.63; P = 0.93), were enhanced with ExT performed in hypoxia versus normoxia. We only found heterogeneity among studies assessing arterial stiffness (I 2 = 63 %, P = 0.02), and no publication bias was detected. Based on current published studies, hypoxic ExT potentiates vascular adaptations related to skeletal muscle capillarization and dilator function. These findings may contribute to establishing effective exercise programs designed to enhance vascular health.


Influence of the BDNF genotype on amygdalo-prefrontal white matter microstructure is linked to nonconscious attention bias to threat.
Cognitive processing biases, such as increased attention to threat, are gaining recognition as causal factors in anxiety. Yet, little is known about the anatomical pathway by which threat biases cognition and how genetic factors might influence the integrity of this pathway, and thus, behavior. For 40 normative adults, we reconstructed the entire amygdalo-prefrontal white matter tract (uncinate fasciculus) using diffusion tensor weighted MRI and probabilistic tractography to test the hypothesis that greater fiber integrity correlates with greater nonconscious attention bias to threat as measured by a backward masked dot-probe task. We used path analysis to investigate the relationship between brain-derived nerve growth factor genotype, uncinate fasciculus integrity, and attention bias behavior. Greater structural integrity of the amygdalo-prefrontal tract correlates with facilitated attention bias to nonconscious threat. Genetic variability associated with brain-derived nerve growth factor appears to influence the microstructure of this pathway and, in turn, attention bias to nonconscious threat. These results suggest that the integrity of amygdalo-prefrontal projections underlie nonconscious attention bias to threat and mediate genetic influence on attention bias behavior. Prefrontal cognition and attentional processing in high bias individuals appear to be heavily influenced by nonconscious threat signals relayed via the uncinate fasciculus.


Lower respiratory tract infections in Inuit infants on Baffin Island.
It has long been suspected that Canadian Inuit children suffer from frequent severe lower respiratory tract infections (LRTIs), but the causes and risk factors have not been documented. This study assessed the infectious causes and other epidemiologic factors that may contribute to the severity of LRTI in young Inuit children on Baffin Island. A prospective case study was carried out at the Baffin Regional Hospital in Iqaluit, Nunavut, of infants less than 6 months of age, who were admitted to hospital between October 1997 and June 1998 with a diagnosis of LRTI. Immunofluorescent antibody testing was used to identify respiratory viruses, and enzyme immunoassay (EIA) and polymerase chain reaction (PCR) were used to test for Chlamydia trachomatis. Demographic and risk factor data were obtained through a questionnaire. The annualized incidence rate of admission to hospital for bronchiolitis at Baffin Regional Hospital was 484 per 1000 infants who were less than 6 months of age; 12% of the infants were intubated. Probable pathogens were identified for 18 of the 27 cases considered in our study. A single agent was identified for 14 infants: 8 had respiratory syncytial virus, 2 adenovirus, 1 rhinovirus, 1 influenza A, 1 parainfluenza 3 and 1 had cytomegalovirus. For 4 infants, 2 infectious agents were identified: these were enterovirus and Bordetella pertussis, adenovirus and enterovirus, cytomegalovirus and respiratory syncytial virus, and respiratory syncytial virus and adenovirus. C. trachomatis was not identified by either EIA or PCR. All infants were exposed to maternal smoking in utero, second-hand smoke at home and generally lived in crowded conditions. Inuit infants in the Baffin Region suffer from an extremely high rate of hospital admissions for LRTI. The high frequency and severity of these infections calls for serious public health attention.