Long-term outcome of percutaneous transluminal angioplasty in chronic haemodialysis patients with peripheral arterial disease.
Chronic haemodialysis patients are at an increased risk of peripheral artery disease (PAD). Although percutaneous transluminal angioplasty (PTA) has become a widely used therapeutic intervention for PAD, its outcome in haemodialysis patients remains poorly understood. The aim of this study was to clarify the long-term outcome of PTA as a primary treatment for PAD in haemodialysis patients. Consecutive 118 haemodialysis patients with 205 lesions and 108 non-haemodialysis patients with 143 lesions who underwent successful PTA as a first-choice therapeutic option for PAD were enrolled. Outcome measures included primary patency, limb salvage and survival. Incidence of diabetes, history of coronary artery disease and femoropopliteal lesion were significantly more frequent in haemodialysis patients (P = 0.008, 0.005 and 0.0001, respectively), but critical limb ischaemia and TransAtlantic Inter-Society Consensus (TASC) lesion types occurred with comparable frequency in both groups. No patients had in-hospital complications. The 5-year primary patency, limb salvage and survival rates were significantly lower in haemodialysis patients (P = 0.01, 0.029 and 0.0024, respectively). On Cox multivariate analysis, haemodialysis was strongly predictive of amputation and all-cause death, but not of restenosis. In haemodialysis patients, TASC C+D lesion and ulceration/gangrene were independent predictors for restenosis and amputation. The long-term outcome after PTA may be fully acceptable in haemodialysis patients who are at the highest risk of cardiovascular disease. PTA is a useful therapeutic strategy in haemodialysis patients with PAD, but PTA for TASC C+D lesions remains controversial.


Perceptions and competence in evidence-based medicine: Are surgeons getting better? A questionnaire survey of members of the Dutch Orthopaedic Association.
The Journal of Bone and Joint Surgery, American Volume (The Journal) recently initiated a section called "Evidence-Based Orthopaedics." Furthermore, a level-of-evidence rating is now used in The Journal to help readers in clinical decision-making. Little is known about whether this recent emphasis has influenced surgeons' perceptions about and competence in evidence-based medicine. Therefore, we examined perceptions and competence in evidence-based medicine among Dutch orthopaedic surgeons. Members of the Dutch Orthopaedic Association were surveyed to examine their attitudes toward evidence-based medicine and their competence in evidence-based medicine. We evaluated competences using a newly developed instrument tailored to surgical practice. Of the 611 members, 367 surgeons (60%) responded. Orthopaedic surgeons welcomed evidence-based medicine. Practical evidence-based medicine resources were perceived as the best method to move from opinion-based or experience-based to evidence-based practice. Four variables were significantly and positively associated with the competence instrument: (1) a younger age, particularly between thirty-six and forty-five years (p = 0.007), (2) experience of less than ten years (p = 0.032), (3) having a PhD degree (p < 0.001), and (4) working in an academic or teaching setting (p = 0.004). The majority of the respondents were aware of The Journal's evidence-based medicine section (84%) and level-of-evidence ratings (65%), and 20% used The Journal's evidence-based medicine abstracts in clinical decision-making. This increased awareness of evidence-based medicine was also reflected in the frequent use of Cochrane reviews in clinical decision-making (27% of the respondents). Surgeons who used and those who were aware of but did not use The Journal's evidence-based medicine abstracts or Cochrane reviews in clinical decision-making had significantly higher competence instrument scores than those who were unaware of these resources (p = 0.03 and p < 0.001, respectively). Evidence-based medicine is welcomed by Dutch orthopaedic surgeons. The recent emphasis on evidence-based medicine is reflected in an increased awareness about The Journal's evidence-based medicine section, levels of evidence, and the largest evidence-based medicine resource: the Cochrane reviews. Younger orthopaedic surgeons had better knowledge about evidence-based medicine. The development and use of evidence-based resources as well as preappraised summaries such as The Journal's evidence-based medicine abstracts and Cochrane reviews were perceived as the best way to move from opinion-based to evidence-based orthopaedic practice.


The National DNA Data Bank of Canada: a Quebecer perspective.
The Canadian National DNA Database was created in 1998 and first used in the mid-2000. Under management by the RCMP, the National DNA Data Bank of Canada offers each year satisfactory reported statistics for its use and efficiency. Built on two indexes (convicted offenders and crime scene indexes), the database not only provides increasing matches to offenders or linked traces to the various police forces of the nation, but offers a memory repository for cold cases. Despite these achievements, the data bank is now facing new challenges that will inevitably defy the way the database is currently used. These arise from the increasing power of detection of DNA traces, the diversity of demands from police investigators and the growth of the bank itself. Examples of new requirements from the database now include familial searches, low-copy-number analyses and the correct interpretation of mixed samples. This paper aims to develop on the original way set in Québec to address some of these challenges. Nevertheless, analytic and technological advances will inevitably lead to the introduction of new technologies in forensic laboratories, such as single cell sequencing, phenotyping, and proteomics. Furthermore, it will not only request a new holistic/global approach of the forensic molecular biology sciences (through academia and a more investigative role in the laboratory), but also new legal developments. Far from being exhaustive, this paper highlights some of the current use of the database, its potential for the future, and opportunity to expand as a result of recent technological developments in molecular biology, including, but not limited to DNA identification.


Risk of Helicobacter pylori infection among long-term residents in developing countries.
The seroprevalence and incidence of Helicobacter pylori infection were determined among 312 North American missionaries who were serving in developing countries between 1967 and 1984. The majority (81%) resided in sub-Saharan Africa. When initially evaluated, the missionaries had a mean age of 40 years, 65% were female, and all were of white race/ethnicity. An ELISA showed that the initial prevalence of IgG antibody to H. pylori was 17%. After a mean of 7.4 years of service (1917 person-years of exposure), 37 (14%) of 259 initially seronegative subjects seroconverted to anti-H. pylori, giving an annual incidence of 1.9%. These data indicate a relatively higher risk of H. pylori infection among missionaries compared with an annual incidence of seroconversion of 0.3-1.0% in industrialized nations. Long-term residents in developing countries should be evaluated for H. pylori infection when gastrointestinal symptoms develop.


Population structure of Stemphylium lycopersici associated with leaf spot of tomato in a single field.
Stemphylium lycopersici is an important pathogen causing leaf spot of tomatoes worldwide. Although much information is available about the pathogen, little is known about dynamics of S. lycopersici in tomato fields. Seventy-nine symptomatic leaf samples were collected from two tomato cultivars grown in a farm (Miral and Inbred line). Fungal species associated with the disease were isolated on potato dextrose agar. Seventy-nine isolates were obtained and identified as S. lycopersici based on sequence analysis of combined dataset of the internal transcribed spacer and glyceraldehyde-3-phosphate dehydrogenase regions. The 79 isolates were subjected to amplified fragment length polymorphism analysis using three primer combinations. The Stemphylium lycopersici population from the two cultivars was found to have a very low level of genetic diversity (H = 0.0948). Cluster analysis showed intermixing of isolates from the two cultivars. In addition, analysis of molecular variance showed the presence of a very low level of genetic differentiation between populations obtained from the two cultivars (Fst = 0.0206). These findings indicate the presence of a high rate of gene flow between the two populations and may suggest that the two populations originated from the same inoculum source. The implications of these findings on the management of Stemphylium-induced leaf spot of tomatoes are discussed.


A Comparison of the Preservation of Mouse Adipose Tissue-Derived Mesenchymal Stem Cells Using the University of Wisconsin Solution and Hank's Balanced Salt Solution.
Preservation of adipose tissue before the isolation of cells is one of the most important steps in maintaining the cell viability of adipose tissue-derived mesenchymal stem cells (ADSCs) for clinical use. Hank's balanced salt solution (HBSS) is one of the main ADSC preservation solutions used clinically. However, this step is known to lead to decreased cell viability. The University of Wisconsin (UW) solution is recognized by transplant physicians as an excellent organ preservation solution. We aimed to investigate the effectiveness of UW solution in preservation of the viability of ADSCs. We collected adipose tissue from the inguinal fat pad of mice and compared preservation in UW solution and HBSS overnight by measuring cell viability after isolation. We found that the number of viable cells harvested per gram of adipose tissue mass was higher in UW solution- than HBSS-preserved tissue.


Differential expression of miRNAs in Brassica napus root following infection with Plasmodiophora brassicae.
Canola (oilseed rape, Brassica napus L.) is susceptible to infection by the biotrophic protist Plasmodiophora brassicae, the causal agent of clubroot. To understand the roles of microRNAs (miRNAs) during the post-transcriptional regulation of disease initiation and progression, we have characterized the changes in miRNA expression profiles in canola roots during clubroot disease development and have compared these to uninfected roots. Two different stages of clubroot development were targeted in this miRNA profiling study: an early time of 10-dpi for disease initiation and a later 20-dpi, by which time the pathogen had colonized the roots (as evident by visible gall formation and histological observations). P. brassicae responsive miRNAs were identified and validated by qRT-PCR of miRNAs and the subsequent validation of the target mRNAs through starBase degradome analysis, and through 5' RLM-RACE. This study identifies putative miRNA-regulated genes with roles during clubroot disease initiation and development. Putative target genes identified in this study included: transcription factors (TFs), hormone-related genes, as well as genes associated with plant stress response regulation such as cytokinin, auxin/ethylene response elements. The results of our study may assist in elucidating the role of miRNAs in post-transcriptional regulation of target genes during disease development and may contribute to the development of strategies to engineer durable resistance to this important phytopathogen.


ClpX stimulates the mitochondrial unfolded protein response (UPRmt) in mammalian cells.
Proteostasis is crucial for life and maintained by cellular chaperones and proteases. One major mitochondrial protease is the ClpXP complex, which is comprised of a catalytic ClpX subunit and a proteolytic ClpP subunit. Based on two separate observations, we hypothesized that ClpX may play a leading role in the cellular function of ClpXP. Therefore, we analyzed the effect of ClpX overexpression on a myoblast proteome by quantitative proteomics. ClpX overexpression results in the upregulation of markers of the mitochondrial proteostasis pathway, known as the "mitochondrial unfolded protein response" (UPRmt). Although this pathway is described in detail in Caenorhabditis elegans, it is not clear whether it is conserved in mammals. Therefore, we compared features of the classical nematode UPRmt with our mammalian ClpX-triggered UPRmt dataset. We show that they share the same retrograde mitochondria-to-nucleus signaling pathway that involves the key UPRmt transcription factor CHOP (also known as Ddit3, CEBPZ or GADD153). In conclusion, our data confirm the existence of a mammalian UPRmt that has great similarity to the C. elegans pathway. Furthermore, our results illustrate that ClpX overexpression is a good and simple model to study the underlying mechanisms of the UPRmt in mammalian cells.


Comprehensive quality evaluation and comparison of Angelica sinensis radix and Angelica acutiloba radix by integrated metabolomics and glycomics.
Angelica radix (Danggui in Chinese) used in China and Japan is derived from two species of Angelica, namely Angelica sinensis and Angelica acutiloba, respectively. The differences in quality between A. sinensis radix (ASR) and A. acutiloba radix (AAR) should be therefore investigated to guide the medicinal and dietary applications of these two species. Secondary metabolites and carbohydrates have been demonstrated to be the two major kinds of bioactive components of Danggui. However, previously, quality comparison between ASR and AAR intensively concerned secondary metabolites but largely overlooked carbohydrates, thus failing to include or take into consideration an important aspect of the holistic quality of Danggui. In this study, untargeted/targeted metabolomics and glycomics were integrated by multiple chromatography-based analytical techniques for qualitative and quantitative characterization of secondary metabolites and carbohydrates in Danggui so as to comprehensively evaluate and compare the quality of ASR and AAR. The results revealed that not only secondary metabolites but also carbohydrates in ASR and AAR were different in type and amount, which should collectively contribute to their quality difference. By providing more comprehensive chemical information, the research results highlighted the need to assess characteristics of both carbohydrates and secondary metabolites for overall quality evaluation and comparison of ASR and AAR.


The Mental Health Outcomes of Drought: A Systematic Review and Causal Process Diagram.
Little is understood about the long term, indirect health consequences of drought (a period of abnormally dry weather). In particular, the implications of drought for mental health via pathways such as loss of livelihood, diminished social support, and rupture of place bonds have not been extensively studied, leaving a knowledge gap for practitioners and researchers alike. A systematic review of literature was performed to examine the mental health effects of drought. The systematic review results were synthesized to create a causal process diagram that illustrates the pathways linking drought effects to mental health outcomes. Eighty-two articles using a variety of methods in different contexts were gathered from the systematic review. The pathways in the causal process diagram with greatest support in the literature are those focusing on the economic and migratory effects of drought. The diagram highlights the complexity of the relationships between drought and mental health, including the multiple ways that factors can interact and lead to various outcomes. The systematic review and resulting causal process diagram can be used in both practice and theory, including prevention planning, public health programming, vulnerability and risk assessment, and research question guidance. The use of a causal process diagram provides a much needed avenue for integrating the findings of diverse research to further the understanding of the mental health implications of drought.


Entomological profile of yellow fever epidemics in the Central African Republic, 2006-2010.
The causative agent of yellow fever is an arbovirus of the Flaviviridae family transmitted by infected Aedes mosquitoes, particularly in Africa. In the Central African Republic since 2006, cases have been notified in the provinces of Ombella-Mpoko, Ouham-Pende, Basse-Kotto, Haute-Kotto and in Bangui the capital. As the presence of a vector of yellow fever virus (YFV) represents a risk for spread of the disease, we undertook entomological investigations at these sites to identify potential vectors of YFV and their abundance. Between 2006 and 2010, 5066 mosquitoes belonging to six genera and 43 species were identified. The 20 species of the Aedes genus identified included Ae. aegypti, the main vector of YFV in urban settings, and species found in tropical forests, such as Ae. africanus, Ae. simpsoni, Ae. luteocephalus, Ae. vittatus and Ae. opok. These species were not distributed uniformly in the various sites studied. Thus, the predominant Aedes species was Ae. aegypti in Bangui (90.7 %) and Basse-Kotto (42.2 %), Ae. africanus in Ombella-Mpoko (67.4 %) and Haute-Kotto (77.8 %) and Ae. vittatus in Ouham-Pende (62.2 %). Ae. albopictus was also found in Bangui. The distribution of these dominant species differed significantly according to study site (P < 0.0001). None of the pooled homogenates of Aedes mosquitoes analysed by polymerase chain reaction contained the YFV genome. The results indicate a wide diversity of vector species for YFV in the Central African Republic. The establishment of surveillance and vector control programs should take into account the ecological specificity of each species.


Recognizing the same face in different contexts: Testing within-person face recognition in typical development and in autism.
Unfamiliar face recognition follows a particularly protracted developmental trajectory and is more likely to be atypical in children with autism than those without autism. There is a paucity of research, however, examining the ability to recognize the same face across multiple naturally varying images. Here, we investigated within-person face recognition in children with and without autism. In Experiment 1, typically developing 6- and 7-year-olds, 8- and 9-year-olds, 10- and 11-year-olds, 12- to 14-year-olds, and adults were given 40 grayscale photographs of two distinct male identities (20 of each face taken at different ages, from different angles, and in different lighting conditions) and were asked to sort them by identity. Children mistook images of the same person as images of different people, subdividing each individual into many perceived identities. Younger children divided images into more perceived identities than adults and also made more misidentification errors (placing two different identities together in the same group) than older children and adults. In Experiment 2, we used the same procedure with 32 cognitively able children with autism. Autistic children reported a similar number of identities and made similar numbers of misidentification errors to a group of typical children of similar age and ability. Fine-grained analysis using matrices revealed marginal group differences in overall performance. We suggest that the immature performance in typical and autistic children could arise from problems extracting the perceptual commonalities from different images of the same person and building stable representations of facial identity.


Preparing Oncology Advanced Practice RNs as Generalists in Palliative Care.
To train and support oncology advanced practice RNs (APRNs) to become generalist providers of palliative care. APRNs with master's or doctor of nursing practice degrees and at least five years of experience in oncology (N = 165) attended a National Cancer Institute-funded national training course and participated in ongoing support and education. Course participants completed a precourse, postcourse, and six-month follow-up evaluation regarding palliative care practices in their settings, course evaluation, and their perceived effectiveness in applying course content in their practice. The precourse results showed deficiencies in current practice, with a low percentage of patients having palliative care as part of their oncology care. Barriers included lack of triggers that could assist in identifying patients who could benefit from palliative care. Six-month postcourse data showed more APRNs participating in family meetings, recommending palliative care consultations, speaking with family members regarding bereavement services, and preparing clinical staff for impending patient deaths. APRNs require palliative care training to integrate this care within their role. APRNs can influence practice change and improve care for patients in their settings.


A semi-quantitative serological method to assess the potency of inactivated rabies vaccine for veterinary use.
Potency is one of the most important indexes of inactivated vaccines. A number of methods have been established to assay the potency, of which the NIH test and single-dose mouse protection test are the "prescribed methods". Here, we report a method to semi-quantitatively assay the potency of an inactivated rabies vaccine, which uses fewer animals and takes less time to complete. Depending on the quality requirements of a vaccine (e.g. minimum potency), a rabies reference vaccine is, for example, diluted to the minimum potency, and 50 μL of the dilution is taken to inoculate 10 mice. The same amount of the test rabies vaccine is inoculated into another 10 mice. After two weeks, all mice are bled and serum samples are assayed for viral neutralizing antibody by the fluorescent antibody virus neutralization (FAVN) test. By comparing the median and interquartile range of antibody titers of the reference vaccine with those of the test vaccine, the test vaccine potency can be semi-quantitatively judged as to whether it is in accord with the required quality. The reliability of this method was also confirmed in dogs. The procedure can be recommended for batch potency testing during inactivated rabies vaccine production.


Comparing the glucuronidation capacity of the feline liver with substrate-specific glucuronidation in dogs.
This study aimed to assess the overall glucuronidation capacity of cats, using prototypic substrates identified for human UDP-glucuronosyltransferases (UGTs). To this end, Michaelis-Menten kinetics were established for the substrates using feline hepatic microsomal fractions, and results were compared with similar experiments carried out with dog liver microsomes. Cats are known for their low capacity of glucuronide formation, and UGT1A6 was found to be a pseudogene. However, functional studies with typical substrates were not performed and knowledge of the enzymology and genetics of other glucuronidation enzymes in felidae is lacking. The results of this study showed extremely low formation of naphthol-1-glucuronide (1.7 ± 0.4 nmol/mg protein/min), estradiol-17-glucuronide (<0.7 nmol/mg protein/min), and morphine-3-glucuronide (0.2 ± 0.03 nmol/mg protein/min), suggesting a lack of functional UGT1A6 and UGT2B7 homologues in the cat's liver. Dog liver microsomes were producing these glucuronides in much higher amounts. Glucuronide capacity was present for the substrates 17β-estradiol (estradiol-3-glucuronide, 2.9 ± 0.2 nmol/mg protein/min) and 4-methylumbelliferone (31.3 ± 3.3 nmol/mg protein/min), assuming that cats have functional homologue enzymes to at least the human UGT1A1 and probably other UGT1A isozymes. This implies that for new drugs, glucuronidation capacity has to be investigated on a substance-to-substance base. Knowledge of the glucuronidation rate of a drug provides the basis for pharmacokinetic modeling and as a result proper dosage regimens can be established to avoid undesirable drug toxicity in cats.


Ideal Soft Tissue Facial Profile in Iranian Males and Females: Clinical Implications.
Proper pre-operative facial analysis that includes a thorough evaluation of both the bony and soft tissue anatomy is paramount to success in performing aesthetic surgery of the face. Ethnic variations in soft tissue profile add an important variable to pre-operative facial analysis. The aim of our study was to determine the role of ethnic variations in soft tissue facial profiles through profile analysis of Iranian male and female patients. Photographs of 100 Iranian males and 100 Iranian females (16 to 40 years old) were carried out. A review committee selected 10 male images and 10 female images, which they believed to be most ideal. The soft tissue profiles were then analyzed. A total of 21 measurements were analyzed and statistically compared with North American Caucasian males and females. The upper lip projection and lower lip projection were significantly more prominent in Iranian males as compared with North American Caucasian males. In addition, Iranian males had longer face as compared with North American Caucasian males along with a more drooping nasal tip. The frontonasal area is straighter and the lower face is longer in Iranian females compared with North American Caucasian in addition to more convex faces along with a shorter upper face. Significant differences in ideal soft tissue profiles exist between Iranian and Caucasian males. These differences should be recognized as they may play an important role in performing facial aesthetic and reconstructive procedures, particularly rhinoplasty, genioplasty, midface/facelifts, lip augmentation, and maxillofacial surgery.


Defining myocardial fibrosis in haemodialysis patients with non-contrast cardiac magnetic resonance.
Extent of myocardial fibrosis (MF) determined using late gadolinium enhanced (LGE) predicts outcomes, but gadolinium is contraindicated in advanced renal disease. We assessed the ability of native T1-mapping to identify and quantify MF in aortic stenosis patients (AS) as a model for use in haemodialysis patients. We compared the ability to identify areas of replacement-MF using native T1-mapping to LGE in 25 AS patients at 3 T. We assessed agreement between extent of MF defined by LGE full-width-half-maximum (FWHM) and the LGE 3-standard-deviations (3SD) in AS patients and nine T1 thresholding-techniques, with thresholds set 2-to-9 standard-deviations above normal-range (1083 ± 33 ms). A further technique was tested that set an individual T1-threshold for each patient (T11SD). The technique that agreed most strongly with FWHM or 3SD in AS patients was used to compare extent of MF between AS (n = 25) and haemodialysis patients (n = 25). Twenty-six areas of enhancement were identified on LGE images, with 25 corresponding areas of discretely increased native T1 signal identified on T1 maps. Global T1 was higher in haemodialysis than AS patients (1279 ms ± 5.8 vs 1143 ms ± 12.49, P < 0.01). No signal-threshold technique derived from standard-deviations above normal-range associated with FWHM or 3SD. T11SD correlated with FWHM in AS patients (r = 0.55) with moderate agreement (ICC = 0.64), (but not with 3SD). Extent of MF defined by T11SD was higher in haemodialysis vs AS patients (21.92% ± 1 vs 18.24% ± 1.4, P = 0.038), as was T1 in regions-of-interest defined as scar (1390 ± 8.7 vs 1276 ms ± 20.5, P < 0.01). There was no difference in the relative difference between remote myocardium and regions defined as scar, between groups (111.4 ms ± 7.6 vs 133.2 ms ± 17.5, P = 0.26). Areas of MF are identifiable on native T1 maps, but absolute thresholds to define extent of MF could not be determined. Histological studies are needed to assess the ability of native-T1 signal-thresholding techniques to define extent of MF in haemodialysis patients. Data is taken from the PRIMID-AS (NCT01658345) and CYCLE-HD studies (ISRCTN11299707).


The global burden of pathogens and pests on major food crops.
Crop pathogens and pests reduce the yield and quality of agricultural production. They cause substantial economic losses and reduce food security at household, national and global levels. Quantitative, standardized information on crop losses is difficult to compile and compare across crops, agroecosystems and regions. Here, we report on an expert-based assessment of crop health, and provide numerical estimates of yield losses on an individual pathogen and pest basis for five major crops globally and in food security hotspots. Our results document losses associated with 137 pathogens and pests associated with wheat, rice, maize, potato and soybean worldwide. Our yield loss (range) estimates at a global level and per hotspot for wheat (21.5% (10.1-28.1%)), rice (30.0% (24.6-40.9%)), maize (22.5% (19.5-41.1%)), potato (17.2% (8.1-21.0%)) and soybean (21.4% (11.0-32.4%)) suggest that the highest losses are associated with food-deficit regions with fast-growing populations, and frequently with emerging or re-emerging pests and diseases. Our assessment highlights differences in impacts among crop pathogens and pests and among food security hotspots. This analysis contributes critical information to prioritize crop health management to improve the sustainability of agroecosystems in delivering services to societies.


Influence of soyabean meal supplementation on the resistance of Scottish blackface lambs to haemonchosis.
Protein supplementation improves the resistance of sheep to haemonchosis. This experiment investigated the Scottish blackface breed to establish whether dietary protein supplementation is still beneficial in a genetically resistant breed. Lambs were given either a basal diet or a diet supplemented with soyabean meal to give an additional 80 g crude protein kg dry matter-1. The lambs were given an initial loading dose of Haemonchus contortus, followed by a trickle infection for 10 weeks. The weight gains of the lambs given the supplemented diet were greater and their carcases were leaner, irrespective of infection status. Infected animals on the basal diet were more anaemic and hypoalbuminaemic than animals receiving the supplemented diet, although there were no statistically significant differences in mean worm burdens or faecal egg counts.


MESOPHYLLUM SPHAERICUM SP. NOV. (CORALLINALES, RHODOPHYTA): A NEW MAËRL-FORMING SPECIES FROM THE NORTHEAST ATLANTIC(1).
Mesophyllum sphaericum sp. nov. is described based on spherical maërl individuals (up to 10 cm) collected in a shallow subtidal maërl bed in Galicia (NW Spain). The thalli of these specimens are radially organized, composed of arching tiers of compact medullary filaments. Epithallial cells have flattened to rounded outermost walls, and they occur in a single layer. Subepithallial initials are as long as, or longer than the daughter cells that subtend them. Cell fusions are abundant. Multiporate asexual conceptacles are protruding, mound-like with a flattened pore plate, lacking a peripheral raised rim. Filaments lining the pore canal and the conceptacle roof are composed of five to six cells with straight elongate and narrow cells at their base. Carposporangial conceptacles are uniporate, protruding, and conical. Spermatangial conceptacles were not observed. Molecular results placed M. sphaericum near to M. erubescens, but M. sphaericum is anatomically close to M. canariense. The examination of the holotype and herbarium specimens of M. canariense indicated that both species have pore canal filaments with elongate basal cells, but they differ in number of cells (five to six in M. sphaericum vs. four in M. canariense). Based on the character of pore canal filaments, M. canariense shows similarities with M. erubescens (three to five celled). The outermost walls of epithallial cells of M. canariense are flared compared to the round to flattened ones of M. erubescens, the latter being widely accepted for the genus Mesophyllum. The addition of M. sphaericum as new maërl-forming species suggests that European maërl beds are more biodiverse than previously understood.


Air quality impacts and health-benefit valuation of a low-emission technology for rail yard locomotives in Atlanta Georgia.
One of the largest rail yard facilities in the Southeastern US, the Inman and Tilford yards, is located in the northwestern section of Atlanta, Georgia alongside other industries, schools, businesses, and dwellings. It is a significant source of fine particulate (PM2.5) and black carbon (BC) (Galvis, Bergin, & Russell, 2013). We calculate 2011 PM2.5 and BC emissions from the rail yards and primary industrial and on-road mobile sources in the area and determine their impact on local air quality using Gaussian dispersion modeling. We determine the change in PM2.5 and BC concentrations that could be accomplished by upgrading traditional switcher locomotives used in these rail yards to a lower emitting technology and evaluate the health benefits for comparison with upgrade costs. Emissions from the rail yards were estimated using reported fuel consumption data (GAEPD, 2012b) and emission factors previously measured in the rail yards (Galvis et al., 2013). Model evaluation against 2011 monitoring data found agreement between measured and simulated concentrations. Model outputs indicate that the line-haul and switcher activities are responsible for increments in annual average concentrations of approximately 0.5±0.03 μg/m(3) (39%) and 0.7±0.04 μg/m(3) (56%) of BC, and for 1.0±0.1 μg/m(3) (7%) and 1.6±0.2 μg/m(3) (14%) of PM2.5 at two monitoring sites located north and south of the rail yards respectively. Upgrading the switcher locomotives at the yards with a lower emitting technology in this case "mother slug" units could decrease PM2.5 and BC emissions by about 9 and 3 t/year respectively. This will lower annual average PM2.5 concentrations between 0.3±0.1 μg/m(3) and 0.6±0.1 μg/m(3) and BC concentrations between 0.1±0.02 μg/m(3) and 0.2±0.03 μg/m(3) at monitoring sites north and south of the rail yards respectively, and would facilitate PM2.5 NAAQS attainment in the area. We estimate that health benefits of approximately 20 million dollars per year could be gained.


"It's the people that make the environment good or bad": the patient's experience of the acute care hospital environment.
A review of contemporary nursing research reveals a tendency to focus on select aspects of the hospital environment such as noise, light, and music. Although studies such as these shed light on discrete aspects of the hospital environment, this body of literature contributes little to an understanding of the entirety of that world as the patient in the sickbed experiences it. The purpose of the study detailed in this article was to describe the patient's experience of the acute care hospital environment. Nondirective, in-depth phenomenological interviews were conducted, then transcribed verbatim, and analyzed for themes. Against the backdrop of "I lived and that's all that matters," there were 3 predominant themes in patients' experience of the acute care environment: (1) disconnection/connection, (2) fear/less fear, and (3) confinement/freedom. In this environment, human-to-human contact increased security and power in an environment that was described as sterile, disorienting, and untrustworthy. Acute and critical care nurses and other caregivers can use the findings to create less noxious hospital environments.


A new reversal mode in exchange coupled antiferromagnetic/ferromagnetic disks: distorted viscous vortex.
Magnetic vortices have generated intense interest in recent years due to their unique reversal mechanisms, fascinating topological properties, and exciting potential applications. In addition, the exchange coupling of magnetic vortices to antiferromagnets has also been shown to lead to a range of novel phenomena and functionalities. Here we report a new magnetization reversal mode of magnetic vortices in exchange coupled Ir20Mn80/Fe20Ni80 microdots: distorted viscous vortex reversal. In contrast to the previously known or proposed reversal modes, the vortex is distorted close to the interface and viscously dragged due to the uncompensated spins of a thin antiferromagnet, which leads to unexpected asymmetries in the annihilation and nucleation fields. These results provide a deeper understanding of the physics of exchange coupled vortices and may also have important implications for applications involving exchange coupled nanostructures.


How to perform EUS-guided tattooing?
Recently, we introduced a series of papers describing on how to perform certain techniques and controversies in EUS. In the first paper, "What should be known before performing EUS examinations, Part I," the authors discussed clinical information and whether other imaging modalities should be needed before embarking in EUS examination. In Part II, some technical controversies on how EUS is performed are discussed from different points of view by providing the relevant available evidence. Herewith, we describe on how to perform EUS-guided fine needle tattooing (FNT) in daily practice. The aim of this paper is to discuss pros and cons for several issues including historical remarks, injecting material, technical approach, and how to perform EUS-FNT including argues in favor and against.


Mapping the Allosteric Action of Antagonists A740003 and A438079 Reveals a Role for the Left Flipper in Ligand Sensitivity at P2X7 Receptors.
P2X7 receptor (P2X7R) activation requires ∼100-fold higher concentrations of ATP than other P2X receptor (P2XR) subtypes. Such high levels are found during cellular stress, and P2X7Rs consequently contribute to a range of pathophysiological conditions. We have used chimeric and mutant P2X7Rs, coupled with molecular modeling, to produce a validated model of the binding mode of the subtype-selective antagonist A438079 at an intersubunit allosteric site. Within the allosteric site large effects on antagonist action were found for point mutants of residues F88A, D92A, F95A, and F103A that were conserved or similar between sensitive/insensitive P2XR subtypes, suggesting that these side-chain interactions were not solely responsible for high-affinity antagonist binding. Antagonist sensitivity was increased with mutations that remove the bulk of side chains around the center of the binding pocket, suggesting that the dimensions of the pocket make a significant contribution to selectivity. Chimeric receptors swapping the left flipper (around the orthosteric site) reduced both ATP and antagonist sensitivity. Point mutations within this region highlighted the contribution of a P2X7R-specific aspartic acid residue (D280) that modeling suggests forms a salt bridge with the lower body region of the receptor. The D280A mutant removing this charge increased ATP potency 15-fold providing a new insight into the low ATP sensitivity of the P2X7R. The ortho- and allosteric binding sites form either side of the β-strand Y291-E301 adjacent to the left flipper. This structural linking may explain the contribution of the left flipper to both agonist and antagonist action.


Multiple approaches to valuation of conservation design and low-impact development features in residential subdivisions.
Residents, developers and civic officials are often faced with difficult decisions about appropriate land uses in and around metropolitan boundaries. Urban expansion brings with it the potential for negative environmental impacts, but there are alternatives, such as conservation subdivision design (CSD) or low-impact development (LID), which offer the possibility of mitigating some of these effects at the development site. Many urban planning jurisdictions across the Midwest do not currently have any examples of these designs and lack information to identify public support or barriers to use of these methods. This is a case study examining consumer value for conservation and low-impact design features in one housing market by using four different valuation techniques to estimate residents' willingness to pay for CSD and LID features in residential subdivisions. A contingent valuation survey of 1804 residents in Ames, IA assessed familiarity with and perceptions of subdivision development and used an ordered value approach to estimate willingness to pay for CSD and LID features. A majority of residents were not familiar with CSD or LID practices. Residents indicated a willingness to pay for most CSD and LID features with the exception of clustered housing. Gender, age, income, familiarity with LID practices, perceptions of attractiveness of features and the perceived effect of CSD and LID features on ease of future home sales were important factors influencing residents' willingness to pay. A hypothetical referendum measured willingness to pay for tax-funded conservation land purchases and estimated that a property tax of around $50 would be the maximum increase that would pass. Twenty-seven survey respondents participated in a subsequent series of experimental real estate negotiations that used an experimental auction mechanism to estimate willingness to pay for CSD and LID features. Participants indicated that clustered housing (with interspersed preserved forest or open space areas), rain gardens, and neighborhood streams with a forested buffer were the features they were most willing to pay for. Participants were not willing to pay for neighborhood streams without buffers. Finally, a spatial hedonic price model using 2093 homes in Ames, IA was used to estimate the effect of public and private open space on housing values. The model indicated that presence of neighborhood association-owned forest and water features as well as proximity to public parks had significant positive effects on housing prices. However, proximity to a public lake had a negative effect on home values. The four methods used in this study include both stated and revealed preference techniques. Although the relative magnitude of value expressed varied, all methods indicated that residents value CSD and LID subdivision features. Subdivision features that included explicit environmental benefits were also consistently preferred over features that did not. Familiarity with alternative designs was an important factor influencing resident willingness to pay for neighborhood features, and developers and civic officials should consider ways to educate citizens about CSD and LID development techniques to increase interest in these designs.


Serum 25-hydroxy vitamin D concentrations are more deficient/insufficient in peritoneal dialysis than haemodialysis patients in a sunny climate.
Research has identified associations between serum 25(OH)D and a range of clinical outcomes in chronic kidney disease and wider populations. The present study aimed to investigate vitamin D deficiency/insufficiency in dialysis patients and the relationship with vitamin D intake and sun exposure. A cross-sectional study was used. Participants included 30 peritoneal dialysis (PD) (43.3% male; 56.87 ± 16.16 years) and 26 haemodialysis (HD) (80.8% male; 63.58 ± 15.09 years) patients attending a department of renal medicine. Explanatory variables were usual vitamin D intake from diet/supplements (IU day(-1) ) and sun exposure (min day(-1) ). Vitamin D intake, sun exposure and ethnic background were assessed by questionnaire. Weight, malnutrition status and routine biochemistry were also assessed. Data were collected during usual department visits. The main outcome measure was serum 25(OH)D (nm). Prevalence of inadequate/insufficient vitamin D intake differed between dialysis modality, with 31% and 43% found to be insufficient (<50 nm) and 4% and 33% found to be deficient (<25 nm) in HD and PD patients, respectively (P < 0.001). In HD patients, there was a correlation between diet and supplemental vitamin D intake and 25(OH)D (ρ = 0.84, P < 0.001) and average sun exposure and 25(OH)D (ρ = 0.50, P < 0.02). There were no associations in PD patients. The results remained significant for vitamin D intake after multiple regression, adjusting for age, gender and sun exposure. The results highlight a strong association between vitamin D intake and 25(OH)D in HD but not PD patients, with implications for replacement recommendations. The findings indicate that, even in a sunny climate, many dialysis patients are vitamin D deficient, highlighting the need for exploration of determinants and consequences.


Gefitinib and erlotinib in metastatic non-small cell lung cancer: a meta-analysis of toxicity and efficacy of randomized clinical trials.
Tyrosine kinase inhibitors (TKIs) targeting the epidermal growth factor receptor (EGFR) have been evaluated in patients with metastatic and advanced non-small cell lung cancer (NSCLC). The U.S. Food and Drug Administration initially granted accelerated approval to gefitinib but subsequently rescinded the authorization. Erlotinib and afatinib are similar compounds approved for the treatment of metastatic NSCLC. The objective of this study was to compare the efficacy and toxicity of erlotinib, gefitinib, and afatinib in NSCLC. We tabulated efficacy variables including overall response rate (ORR), progression-free survival (PFS), and overall survival (OS) and quantitated toxicities and rates of dose reductions and discontinuation. Summary odds ratios were calculated using random and fixed-effects models. An odds ratio was the summary measure used for pooling of studies. We examined 28 studies including three randomized trials with afatinib. Clinical toxicities, including pruritus, rash, anorexia, diarrhea, nausea, fatigue, mucositis, paronychia, and anemia, were similar between erlotinib and gefitinib, although some statistical differences were observed. Afatinib treatment resulted in more diarrhea, rash, and paronychia compared with erlotinib and gefitinib. Regarding efficacy, similar outcomes were recorded for ORR, PFS, or OS in the total population and in specific subgroups of patients between erlotinib and gefitinib. All three TKIs demonstrated higher ORRs in first line in tumors harboring EGFR mutations. Gefitinib has similar activity and toxicity compared with erlotinib and offers a valuable alternative to patients with NSCLC. Afatinib has similar efficacy compared with erlotinib and gefitinib in first-line treatment of tumors harboring EGFR mutations but may be associated with more toxicity, although further studies are needed. Gefitinib deserves consideration for U.S. marketing as a primary treatment for EGFR-mutant NSCLC.


Short communication: Farm and socioeconomic characteristics of the top 100 dairy farm counties in the United States.
The objectives of this study are to describe dairy farm demographic and socioeconomic conditions in the top 100 counties in the United States for dairy sales in 2007, and to describe the association of dairy farm demographics with socioeconomic conditions. The top 100 counties were responsible for 56% of all US dairy sales in 2007 with a median growth rate of 78% compared with 1997. Counties varied widely for farm demographics with as few as 5 very large dairy farms that averaged $17,924,000 in dairy sales per farm to as many as 1,730 dairy farms with less than $250,000 in dairy sales per farm. Most of the top 100 dairy counties had higher illiteracy rates, a higher proportion of residents without a high school degree, and lower median incomes than state averages, but unemployment rates were similar to the state average. The socioeconomic measures were from public records and not collected specifically for this research. Nevertheless, the top dairy counties in the western states tended to have poorer socioeconomic conditions than the top dairy counties in other regions, and significant associations were observed between dairy farm demographics and socioeconomic conditions. Having many dairy farms was associated more favorably with county socioeconomic conditions than having high dairy sales.


Trace copper(II) or zinc(II) ions drastically modify the aggregation behavior of amyloid-beta1-42: an AFM study.
Formation of amyloid-beta (Abeta){1-42} amyloid fibrils, a characteristic feature of Alzheimer's disease (AD), was monitored in situ through atomic force microscopy (AFM). Well-structured amyloid fibrils slowly formed in solution within 24 hours for which high quality AFM pictures could be obtained. Remarkably, addition of either copper(II) or zinc(II) ions to the incubation medium, even at extremely low molar ratios, dramatically changed the Abeta {1-42} aggregation profile and prevented fibril formation. Aggregates of different morphology appeared in accordance with previous observations: small globular aggregates upon addition of zinc; ill-structured micro-aggregates in the case of copper. The implications of these AFM results are discussed in the context of current concepts for AD metallobiology.


Biofeedback for pain management during labour.
Labour is often associated with pain and discomfort caused by a complex and subjective interaction of multiple factors, and should be understood within a multi-dimensional and multi-disciplinary framework. Within the non-pharmacological approach, biofeedback has focused on the acquisition of control over some physiological responses with the aid of electronic devices, allowing individuals to regulate some physical processes (such as pain) which are not usually under conscious control. The role of this behavioural approach for the management of pain during labour, as an addition to the standard prenatal care, has been never assessed systematically. This review is one in a series of Cochrane reviews examining pain relief in labour, which will contribute to an overview of systematic reviews of pain relief for women in labour (in preparation). To examine the effectiveness of the use of biofeedback in prenatal lessons for managing pain during labour. We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (31 March 2011), CENTRAL (The Cochrane Library 2011, Issue 1), PubMed (1950 to 20 March 2011), EMBASE (via OVID) (1980 to 24 March 2011), CINAHL (EBSCOhost) (1982 to 24 March 2011), and PsycINFO (via Ovid) (1806 to 24 March 2011). We searched for further studies in the reference lists of identified articles. Randomised controlled trials of any form of prenatal classes which included biofeedback, in any modality, in women with low-risk pregnancies. Two authors independently assessed trial quality and extracted data. The review included four trials (186 women) that hugely differed in terms of the diversity of the intervention modalities and outcomes measured. Most trials assessed the effects of electromyographic biofeedback in women who were pregnant for the first time. The trials were judged to be at a high risk of bias due to the lack of data describing the sources of bias assessed. There was no significant evidence of a difference between biofeedback and control groups in terms of assisted vaginal birth, caesarean section, augmentation of labour and the use of pharmacological pain relief. The results of the included trials showed that the use of biofeedback to reduce the pain in women during labour is unproven. Electromyographic biofeedback may have some positive effects early in labour, but as labour progresses there is a need for additional pharmacological analgesia. Despite some positive results shown in the included trials, there is insufficient evidence that biofeedback is effective for the management of pain during labour.


Novel irreversible fluorescent probes targeting the 18 kDa translocator protein: synthesis and biological characterization.
The 18 kDa translocator protein (TSPO) is a mitochondrial protein whose basal density is altered in several diseases, with the result that the evaluation of its expression levels by means of molecular imaging techniques represents a promising diagnostic approach. Experimental procedures using a labeled ligand often cause loss of the bound probe, and consequently high affinity ligands covalently binding the receptor protein are needed to overcome this problem. We have previously described a series of N,N-dialkyl-(2-phenylindol-3-yl)glyoxylamides as potent and selective TSPO ligands. Starting from these derivatives, we designed novel TSPO irreversible ligands bearing an electrophilic isothiocyanato group (7, 8), together with an irreversible NBD-fluorescent probe (18). The TSPO affinity of the new irreversible ligands was measured on rat tissue homogenates by [(3)H]Ro 5-4864 radiobinding kinetic assays, all compounds showing high affinities for the target protein. Further biological characterization of the fluorescent irreversible TSPO probe 18 was carried out by using fluorescent spectroscopy in human glioma cells.


Effects of wearing gumboots and leather lace-up boots on lower limb muscle activity when walking on simulated underground coal mine surfaces.
This study aimed to investigate the effects of wearing two standard underground coal mining work boots (a gumboot and a leather lace-up boot) on lower limb muscle activity when participants walked across simulated underground coal mining surfaces. Quadriceps (rectus femoris, vastus medialis, vastus lateralis) and hamstring (biceps femoris, semitendinosus) muscle activity were recorded as twenty male participants walked at a self-selected pace around a circuit while wearing each boot type. The circuit consisted of level, inclined and declined surfaces composed of rocky gravel and hard dirt. Walking in a leather lace-up boot, compared to a gumboot, resulted in increased vastus lateralis and increased biceps femoris muscle activity when walking on sloped surfaces. Increased muscle activity appears to be acting as a slip and/or trip prevention strategy in response to challenging surfaces and changing boot features.


First human ventral mesencephalon and striatum cografting in a Parkinson patient.
Fetal ventral mesencephalon (VM) transplantation has been reported to improve parkinsonian symptoms. Animal studies show that cografting of striatal tissue increases the survival of dopamine neurons. Whether or not VM and striatum cografting could ameliorate motor dysfunction in a Parkinson's disease (PD) patient was explored in this study. The patient was a 53-year-old male who had presented with symptoms of tremor, rigidity and bradykinesia for 11 years. He had been treated with L-dopa and had progressive deterioration of symptoms even with the daily dosage of L-dopa increased to 900 mg per day. Before transplantation, his PD symptoms were scored with Unified Parkinson's Disease Rating Scale (UPDRS) and video recordings. The influx constant (ki) of the [18F] 6-fluoro-L-dopa uptake in the striatum was measured by positron emission tomography (PET) imaging. The fetal VM and the lateral part of the lateral ganglionic eminence (LGE) were cografted into the right putamen and, one week later, fetal VM alone was transplanted into the left putamen. After the transplantation, the patient's UPDRS score improved from 128 to 62 at 6 months and to 24 at 22 months during the "off" phase. The score of daily living disability improved from 35 to 18 at 6 months and to 10 at 22 months post transplantation. Twenty-two months after grafting, "off" phases were almost absent, and the freezing had totally disappeared. The [18F] 6-fluoro-L-dopa PET studies were performed 1 month before and 21 months after transplantation. The ki for [18F] 6-fluoro-L-dopa was decreased by 15% in the right caudate and 5% in the left caudate, both of which did not have any ventral mesencephalic grafts. However, the ki was increased by 35%, in the left non-cografted putamen, and by 58% in the right cografted putamen. In conclusion, cografting the fetal VM and the LGE in the putamen may improve the motor function of PD patients.


An Essential Service Decision Model for ABA Providers During Crisis.
In the United States, applied behavior analysis (ABA) is broadly recognized as a medically necessary treatment for individuals diagnosed with autism and related disorders (Association of Professional Behavior Analysts, 2020, Guidelines for practicing applied behavior analysis during COVID-19 pandemic, Retrieved from https://cdn.ymaws.com/www.apbahome.net/resource/collection/1FDDBDD2-5CAF-4B2A-AB3F-DAE5E72111BF/APBA_Guidelines_-_Practicing_During_COVID-19_Pandemic_040920.pdf). We argue that this designation should not be called into question in light of a particular disaster and that it is critical to consider that an interruption of services can have long-lasting effects on the treatment of the individual (practitioners are ethically obligated to uphold the continuity of services while doing no harm). This dilemma might be ameliorated by a decision model that considers the prioritization of immediate needs, the vulnerability of clients, and the competency of service providers. Just as the medical field prioritizes immediate needs during crisis situations and defers routine appointments (e.g., physicals, checkups), the ABA field can make similar evidence-based decisions. The purpose of the current article is to provide a decision model for ABA practitioners who find themselves questioning the need for essential service delivery during the current COVID-19 pandemic. The impact of this model goes beyond the needs of this crisis and can be applied to any emergency situation where services are at risk of interruption.


The insulinotropic potency of fatty acids is influenced profoundly by their chain length and degree of saturation.
Lowering of the elevated plasma FFA concentration in 18- 24-h fasted rats with nicotinic acid (NA) caused complete ablation of subsequent glucose-stimulated insulin secretion (GSIS). Although the effect of NA was reversed when the fasting level of total FFA was maintained by coinfusion of soybean oil or lard oil (plus heparin), the more saturated animal fat proved to be far more potent in enhancing GSIS. We therefore examined the influence of individual fatty acids on insulin secretion in the perfused rat pancreas. When present in the perfusion fluid at 0.5 mM (in the context of 1% albumin), the fold stimulation of insulin release from the fasted pancreas in response to 12.5 mM glucose was as follows: octanoate (C8:0), 3.4; linoleate (C18:2 cis/cis), 5.3; oleate (C18:1 cis), 9.4; palmitate (C16:0), 16. 2; and stearate (C18:0), 21.0. The equivalent value for palmitoleate (C16:1 cis) was 3.1. A cis--> trans switch of the double bond in the C16:1 and C18:1 fatty acids had only a modest, if any, impact on their potency. A similar profile emerged with regard to basal insulin secretion (3 mM glucose). When a subset of these fatty acids was tested in pancreases from fed animals, the same rank order of effectiveness at both basal and stimulatory levels of glucose was seen. The findings reaffirm the essentiality of an elevated plasma FFA concentration for GSIS in the fasted rat. They also show, however, that the insulinotropic effect of individual fatty acids spans a remarkably broad range, increasing and decreasing dramatically with chain length and degree of unsaturation, respectively. Thus, for any given level of glucose, insulin secretion will be influenced greatly not only by the combined concentration of all circulating (unbound) FFA, but also by the makeup of this FFA pool. Both factors will likely be important considerations in understanding the complex interplay between the nature of dietary fat and whole body insulin, glucose, and lipid dynamics.


Polymorphous hemangioendothelioma of the neck.
Polymorphous hemangioendotheliomas are rare, low-grade borderline malignant vascular tumors of endothelial cell origin. To the best of our knowledge (MEDLINE 1966-2002), there have been nine cases of polymorphous hemangioendothelioma reported in the English literature. Most of the initial patients reported were men, but we present the third case in a woman. Her previous radiation history to the neck makes this report unique. Polymorphous hemangioendothelioma is characterized by the variety of patterns of growth within and between tumors, making histologic recognition of the tumor difficult. Because management remains conservative via wide local excision, the misdiagnosis of this lesion as a malignancy has possible treatment implications. Alternatively, the high propensity for local recurrence underscores the necessity for accurate classification of the neoplasm and close clinical follow-up.


Three-dimensional multiwaveguide probe array for light delivery to distributed brain circuits.
To deliver light to the brain for neuroscientific and neuroengineering applications like optogenetics, in which light is used to activate or silence neurons expressing specific photosensitive proteins, optical fibers are commonly used. However, an optical fiber is limited to delivering light to a single target within the 3D structure of the brain. Here, we describe the design and fabrication of an array of thin microwaveguides, which terminates at a three-dimensionally distributed set of points, appropriate for delivering light to targets distributed in a 3D pattern throughout the brain.


Tissue resolved, gene structure refined equine transcriptome.
Transcriptome interpretation relies on a good-quality reference transcriptome for accurate quantification of gene expression as well as functional analysis of genetic variants. The current annotation of the horse genome lacks the specificity and sensitivity necessary to assess gene expression especially at the isoform level, and suffers from insufficient annotation of untranslated regions (UTR) usage. We built an annotation pipeline for horse and used it to integrate 1.9 billion reads from multiple RNA-seq data sets into a new refined transcriptome. This equine transcriptome integrates eight different tissues from 59 individuals and improves gene structure and isoform resolution, while providing considerable tissue-specific information. We utilized four levels of transcript filtration in our pipeline, aimed at producing several transcriptome versions that are suitable for different downstream analyses. Our most refined transcriptome includes 36,876 genes and 76,125 isoforms, with 6474 candidate transcriptional loci novel to the equine transcriptome. We have employed a variety of descriptive statistics and figures that demonstrate the quality and content of the transcriptome. The equine transcriptomes that are provided by this pipeline show the best tissue-specific resolution of any equine transcriptome to date and are flexible for several downstream analyses. We encourage the integration of further equine transcriptomes with our annotation pipeline to continue and improve the equine transcriptome.


Cross-species chromosome painting in the golden mole and elephant-shrew: support for the mammalian clades Afrotheria and Afroinsectiphillia but not Afroinsectivora.
Cross-species painting (fluorescence in situ hybridization) with 23 (human Homo sapiens (HSA)) chromosome-specific painting probes (HSA 1-22 and the X) was used to delimit regions of homology on the chromosomes of the golden mole (Chrysochloris asiaticus) and elephant-shrew (Elephantulus rupestris). A cladistic interpretation of our data provides evidence of two unique associations, HSA 1/19p and 5/21/3, that support Afrotheria. The recognition of HSA 5/3/21 expands on the 3/21 synteny originally designated as an ancestral state for all eutherians. We have identified one adjacent segment combination (HSA2/8p/4) that is supportive of Afroinsectiphillia (aardvark, golden mole, elephant-shrew). Two segmental combinations (HSA 10q/17 and HSA 3/20) unite the aardvark and elephant-shrews as sister taxa. The finding that segmental syntenies in evolutionarily distant taxa can improve phylogenetic resolution suggests that they may be useful for testing sequence-based phylogenies of the early eutherian mammals. They may even suggest clades that sequence trees are not recovering with any consistency and thus encourage the search for additional rare genomic changes among afrotheres.


Agonist- and voltage-gated calcium entry in cultured mouse spinal cord neurons under voltage clamp measured using arsenazo III.
Spinal cord neurons is dissociated cell culture were loaded with the calcium indicator arsenazo III using the whole-cell patch-clamp recording technique. Under voltage-clamp, depolarizing voltage steps evoked transient increases in absorbance at 660 nm, with no change at 570 nm, the isosbestic wavelength for calcium-arsenazo III complexes. The optical response occurred with a threshold depolarization to -30 mV, peaked at +10 mV, and decreased with further depolarization, consistent with an elevation of cytoplasmic free calcium resulting from Ca2+ flux through voltage-dependent calcium channels. Inward current responses to the excitatory amino acids N-methyl-D-aspartic acid (NMDA) and L-glutamate were also accompanied by calcium transients; these were dose-dependent, varied with the driving force for inward current, and were blocked by extracellular Mg2+ in a voltage-dependent manner, suggesting Ca2+ flux through NMDA-receptor channels. Responses to kainate, quisqualate, and GABA were not accompanied by comparable calcium transients. [Ca2+]i transients evoked by depolarizing voltage steps were of maximal amplitude at the start of recording and declined with time, reflecting rundown of voltage-dependent calcium channels. In contrast, [Ca2+]i transients evoked by NMDA gradually increased in amplitude during periods of whole-cell recording lasting 1-2 hr. Procedures resulting in loading of the neuron with Ca2+ accelerated the increase in amplitude of [Ca2+]i transients evoked by NMDA, but slowed the decay of [Ca2+]i transients evoked by voltage steps. Our results provide evidence for 2 independent sources of transmembrane Ca2+ flux in vertebrate neurons, through voltage-gated calcium channels and through NMDA-receptor channels. The Ca2+ flux gated by NMDA-receptor-specific agonists may play a role in synaptic plasticity, in regulating excitability, and in the excitotoxic response to excitatory amino acids.


An Open-Label, Single-Arm, Two-Stage, Multicenter, Phase II Study to Evaluate the Efficacy of TLC388 and Genomic Analysis for Poorly Differentiated Neuroendocrine Carcinomas.
The discovery of effective therapeutic options for treating metastatic poorly differentiated neuroendocrine carcinoma (NEC) after prior platinum-based chemotherapy remains elusive. This study analyzed the efficacy of TLC388 (Lipotecan) Hydrochloride, a novel camptothecin analog, for pretreated patients with metastatic NEC. This single-arm, two-stage, phase II clinical trial was conducted at four community and academic centers in Taiwan. Patients aged 20 years or older with confirmed metastatic NEC and who had received prior systemic therapy with etoposide plus cisplatin were enrolled between July 2015 and May 2018. Patients received 40 mg/m2 of TLC388 intravenously on days 1, 8, and 15 of a 28-day cycle until disease progression or unacceptable toxic effects. Gene mutations were analyzed by next-generation sequencing. Twenty-three patients with a median age of 61 (range, 44-73) years, 18 of whom were men (78%), were enrolled. Patients received a median of 2 (range, 0-6) treatment cycles. Among 20 evaluable patients, 3 patients exhibited stable disease and no patient experienced a complete or partial remission, resulting in a disease control rate of 15%. Median progression-free survival was 1.8 (95% confidence interval [CI], 0.4-15) months, and the median overall survival was 4.3 (95% CI, 1.7-15) months. The most common treatment-related hematologic adverse events at grade 3 or higher were leukopenia (22.7%), anemia (31.8%), and thrombocytopenia (18.2%). The most frequent mutated genes in 35 patients with NEC were ARSA, DPYD, HEXB, BRCA1, HPD, MYBPC3, BBS2, IL7R, HSD17B4, and PRODH. TLC388 demonstrates limited antitumor activity in metastatic NEC. ClinicalTrials.gov identifier: NCT02457273. Poorly differentiated neuroendocrine carcinomas (NECs) are rare and aggressive. Currently, effective therapeutic options for treating metastatic poorly differentiated NECs beyond platinum-based chemotherapy remain elusive. In this single-arm, multicenter, phase II study, 23 patients with NEC were enrolled and received TLC388 (Lipotecan) Hydrochloride, which is a novel camptothecin analog. The results demonstrated the disease control rate of 15%, the median progression-free survival of 1.8 (95% confidence interval [CI], 0.4-15) months, and the median overall survival of 4.3 (95% CI, 1.7-15) months. Most importantly, several novel genetic mutations and pathways were identified. These results offer the opportunity to develop future treatment strategies in this rare cancer.


Relation between lipopolysaccharide-induced endothelial cell injury and entry of macromolecules into the rat aorta in vivo.
Lipopolysaccharide (LPS) causes endothelial cell injury both in vitro and in vivo. It is widely believed that this injury in vivo enhances the transport of macromolecules from plasma into the interstitial space of the underlying artery wall. A new technique was used in rats to obtain high resolution transmural profiles of macromolecules in vivo. We compared the time course of the macromolecular transport into the aortic tissue in vivo after LPS injection to that of LPS-induced endothelial cell death and the proliferative response of the endothelium to LPS injury. At a dose of 1 mg LPS/kg body wt, endothelial cell death reached a maximum by 36 hours after LPS injection and remained elevated for 96 hours; the peak of the S phase of endothelial cell proliferation was observed 48 hours after injection. To examine the effect of LPS on macromolecular accumulation, we measured aortic intimal and medial transmural concentration profiles of horseradish peroxidase (HRP) after circulation of HRP for 15 minutes. The data revealed a transient increase in total aortic accumulation (reflecting predominantly the media), which was maximal between 12 and 48 hours after LPS injection. Although total medial accumulation was found to return to near control levels by 72 hours after LPS injection, intimal accumulation remained elevated above control levels for 120 hours. When HRP was added to the perfusate of an in situ aorta preparation at a near zero transmural pressure gradient, the resulting transmural concentration profiles across aortas from control rats and from rats given LPS 24 hours previously were indistinguishable, whereas a pressure gradient of 60 mm Hg revealed LPS-altered concentration profiles analogous to those in vivo. This suggests that the accumulation of HRP observed in vivo was driven by increased convective transport. These results reveal that LPS enhances entry of macromolecules into the aorta wall in vivo. The changes in macromolecular transport do not, however, correlate temporally with endothelial cell death or proliferation. The results are consistent with an LPS-induced decrease in the endothelial barrier function, which precedes, and may be independent of, cell death and a transient increase in convective transport across the media due to alterations in the barrier function of the internal elastic lamina.


Hybridization-based detection of Helicobacter pylori at human body temperature using advanced locked nucleic acid (LNA) probes.
The understanding of the human microbiome and its influence upon human life has long been a subject of study. Hence, methods that allow the direct detection and visualization of microorganisms and microbial consortia (e.g. biofilms) within the human body would be invaluable. In here, we assessed the possibility of developing a variant of fluorescence in situ hybridization (FISH), named fluorescence in vivo hybridization (FIVH), for the detection of Helicobacter pylori. Using oligonucleotide variations comprising locked nucleic acids (LNA) and 2'-O-methyl RNAs (2'OMe) with two types of backbone linkages (phosphate or phosphorothioate), we were able to successfully identify two probes that hybridize at 37 °C with high specificity and sensitivity for H. pylori, both in pure cultures and in gastric biopsies. Furthermore, the use of this type of probes implied that toxic compounds typically used in FISH were either found to be unnecessary or could be replaced by a non-toxic substitute. We show here for the first time that the use of advanced LNA probes in FIVH conditions provides an accurate, simple and fast method for H. pylori detection and location, which could be used in the future for potential in vivo applications either for this microorganism or for others.


Terahertz thermometry of gold nanospheres in water.
The photo-thermal effects of plasmonic nanoparticles are promising for cancer therapies. These treatments would greatly benefit from real-time, multi-scale temperature mapping by non-invasive means. Here we show that intense terahertz time domain spectroscopy can be used as a non-contact and high-resolution thermometer of water solutions. Using this technique, we measure the temperature change, triggered by femtosecond amplified laser pulses, of a solution of gold nanospheres in water. Extensions of this ultra-fast and non-invasive technique could open the door to real-time micro-thermometry of single cells without fluorescent labels.


Core Competencies for Interprofessional Collaborative Practice Among Teacher Education, Health and Social Care Students in a Large Scaled Blended Learning Course.
Society's demands for better coordination of services for children are increasing. Interprofessional learning (IPL) has been suggested to achieve the triple aim of better services, better outcomes and reduced costs. The aims were to assess 1) to what extent students taking teacher education, health and social care programmes agreed that blended learning was a suitable approach in a mandatory IPL course, 2) to what extent they had learnt about the WHO's core IPL competencies (roles and responsibilities, values and ethics, interprofessional communication, and teams and teamwork), and 3) the students' ranking of the learning outcomes from different components of the IPL course. This was a quantitative cross-sectional study. Students completed an online course evaluation after a two-day combination of online and face-to-face IPL small-group training. The response rate was 25.8% (n=363). Among the students, 60.6% strongly agreed that blended learning was suitable, while 8.9% strongly disagreed. Among the respondents, 46.8%, 50.2%, 56.8% and 62.3% gained increased insight into roles and responsibilities, values and ethics, interprofessional communication, and teams and teamwork, respectively. In ascending order, students were most satisfied with the learning outcomes from the supervision (16.0%), the syllabus (28.6%), the submission assignment (42.4%), the digital learning content of Canvas (43.8%), the combination of everything (43.8%), and the IPL group discussions (78.6%). In stratified analyses, 'teacher education and child welfare students' were significantly more likely to gain better insight into the WHO competencies than "health and social care students", and they were also more overall satisfied. Students agreed that blended learning was a suitable approach, although the learning outcomes from the face-to-face discussions were markedly higher than from other course components. While the majority had learnt something about the WHO competencies, the teacher and child welfare students achieved the best learning outcomes, including new knowledge about the WHO competencies.


Estimation of central aortic pressure waveform by mathematical transformation of radial tonometry pressure. Validation of generalized transfer function.
Central aortic pressures and waveform convey important information about cardiovascular status, but direct measurements are invasive. Peripheral pressures can be measured noninvasively, and although they often differ substantially from central pressures, they may be mathematically transformed to approximate the latter. We tested this approach, examining intersubject and intrasubject variability and the validity of using a single averaged transformation, which would enhance its applicability. Invasive central aortic pressure by micromanometer and radial pressure by automated tonometry were measured in 20 patients at steady state and during hemodynamic transients (Valsalva maneuver, abdominal compression, nitroglycerin, or vena caval obstruction). For each patient, transfer functions (TFs) between aortic and radial pressures were calculated by parametric model and results averaged to yield individual TFs. A generalized TF was the average of individual functions. TFs varied among patients, with coefficients of variation for peak amplitude and frequency at peak amplitude of 24.9% and 16.9%, respectively. Intrapatient TF variance with altered loading (> 20% variation in peak amplitude) was observed in 28.5% of patients. Despite this, the generalized TF estimated central arterial pressures to < or = 0.2 +/- 3.8 mm Hg error, arterial compliance to 6 +/- 7% accuracy, and augmentation index to within -7% points (30 +/- 45% accuracy). Individual TFs were only marginally superior to the generalized TF for reconstructing central pressures. Central aortic pressures can be accurately estimated from radial tonometry with the use of a generalized TF. The reconstructed waveform can provide arterial compliance estimates but may underestimate the augmentation index because the latter requires greater fidelity reproduction of the wave contour.


Racial/ethnic disparities in HIV infection among people who inject drugs: an international systematic review and meta-analysis.
The Ethnic Minority Meta-Analysis (EMMA) aims to assess racial/ethnic disparities in HIV infection among people who inject drugs (PWID) across various countries. This is the first report of the data. Standard systematic review/meta-analysis methods were utilized, including searching for, screening and coding published and unpublished reports and meta-analytical statistics. We followed the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) statement and Meta-analysis of Observational Studies in Epidemiology (MOOSE) guidelines for reporting methods. Disparities were measured with the odds ratio (OR) for HIV prevalence among ethnic minority PWID compared to ethnic majority PWID; an OR >1.0 indicated higher prevalence among ethnic minorities. Racial/ethnic disparities in HIV prevalence among PWID were examined in 131 prevalence reports, with 214 racial/ethnic minority to majority comparisons, comprising 106 715 PWID. Overall, the pooled OR indicates an increased likelihood of higher HIV prevalence among racial/ethnic minority compared to racial/ethnic majority PWID [OR = 2.09, 95% confidence interval (CI): 1.92-2.28]. Among 214 comparisons, 106 produced a statistically significant higher OR for minorities; in 102 comparisons the OR was not significantly different from 1.0; six comparisons produced a statistically significant higher OR for majority group members. Disparities were particularly large in the United States, pooled OR = 2.22 (95% CI: 2.03-2.44). There was substantial variation in ORs-I(2) = 75.3%: interquartile range = 1.38-3.56-and an approximate Gaussian distribution of the log ORs. Among people who inject drugs, ethnic minorities are approximately twice as likely to be HIV seropositive than ethnic majorities. The great heterogeneity and Gaussian distribution suggest multiple causal factors and a need to tailor interventions to local conditions.


Promoting gross motor skills and physical activity in childcare: A translational randomized controlled trial.
Educator-led programs for physical activity and motor skill development show potential but few have been implemented and evaluated using a randomized controlled design. Furthermore, few educator-led programs have evaluated both gross motor skills and physical activity. Therefore, the aim of this study was to evaluate a gross motor skill and physical activity program for preschool children which was facilitated solely by childcare educators. A six-month 2-arm randomized controlled trial was implemented between April and September 2012 in four early childhood centers in Tasmania, Australia. Educators participated in ongoing professional development sessions and children participated in structured physical activity lessons and unstructured physical activity sessions. In total, 150 children were recruited from four centers which were randomized to intervention or wait-list control group. Six early childhood educators from the intervention centers were trained to deliver the intervention. Gross motor skills were assessed using the Test of Gross Motor Development (2nd edition) and physical activity was measured objectively using GT3X+ Actigraph accelerometers. No statistically significant differences were identified. However, small to medium effect sizes, in favor of the intervention group, were evident for four of the five gross motor skills and the total gross motor skill score and small to medium effect sizes were reported for all physical activity outcomes. This study highlights the potential of educator-led physical activity interventions and supports the need for further translational trials within the early childhood sector.


Ventilatory sensitivity to carbon dioxide before and after episodic hypoxia in women treated with testosterone.
We hypothesized that the ventilatory threshold and sensitivity to carbon dioxide in the presence of hypoxia and hyperoxia during wakefulness would be increased following testosterone administration in premenopausal women. Additionally, we hypothesized that the sensitivity to carbon dioxide increases following episodic hypoxia and that this increase is enhanced after testosterone administration. Eleven women completed four modified carbon dioxide rebreathing trials before and after episodic hypoxia. Two rebreathing trials before and after episodic hypoxia were completed with oxygen levels sustained at 150 Torr, the remaining trials were repeated while oxygen was maintained at 50 Torr. The protocol was completed following 8-10 days of treatment with testosterone or placebo skin patches. Resting minute ventilation was greater following treatment with testosterone compared with placebo (testosterone 11.38 +/- 0.43 vs. placebo 10.07 +/- 0.36 l/min; P < 0.01). This increase was accompanied by an increase in the ventilatory sensitivity to carbon dioxide in the presence of sustained hyperoxia (VSco(2)(hyperoxia)) compared with placebo (3.6 +/- 0.5 vs. 2.9 +/- 0.3; P < 0.03). No change in the ventilatory sensitivity to carbon dioxide in the presence of sustained hypoxia (VSco(2 hypoxia)) following treatment with testosterone was observed. However, the VSco(2 hypoxia) was increased after episodic hypoxia. This increase was similar following treatment with placebo or testosterone patches. We conclude that treatment with testosterone leads to increases in the VSco(2)(hyperoxia), indicative of increased central chemoreflex responsiveness. We also conclude that exposure to episodic hypoxia enhances the VSco(2 hypoxia), but that this enhancement is unaffected by treatment with testosterone.


A clinically translatable mouse model for chemotherapy-related fatigue.
Fatigue is a debilitating and pervasive complication of cancer and cancer care. Clinical research investigating potential therapies is hindered by variability in patient histories, different metrics for measuring fatigue, and environmental factors that may affect fatigue. The purpose of this study was to establish an animal model of chemotherapy-related fatigue. Female HSD:ICR mice were treated with doxorubicin (2.5 mg/kg) or saline in 2 cycles (days 1 through 3 and 10 through 12). After treatment, mice were individually housed in cages equipped with running wheels. Open-field activity and motor coordination were examined after each cycle of treatment and after each week of wheel running. In a separate cohort, modafinil (50 mg/kg) was assessed as a potential treatment for fatigue. Doxorubicin administration resulted in greater than 30% less wheel running compared with that of saline controls. Activity differences were specific to wheel running: neither distance traveled in the open field nor motor coordination according to the rotarod test differed between groups. Compared with control values, RBC counts in the doxorubicin group were decreased on days 15 and 22 but recovered to control levels by study completion. Modafinil was efficacious in increasing wheel running in the doxorubicin group. The current results establish an animal model of chemotherapy-related fatigue that recapitulates the physical symptoms of cancer-related fatigue as manifested as decreased voluntary activity. This model is sensitive to pharmaceutical intervention and can be used to screen potential treatments for fatigue.


Effect of fitness on glucose, insulin and cortisol responses to diets varying in starch and fat content in Thoroughbred horses with recurrent exertional rhabdomyolysis.
Recurrent exertional rhabdomyolysis (RER) occurs in fit, nervous Thoroughbreds fed high nonstructural carbohydrate (NSC) diets. Clinical signs are diminished by feeding low NSC, high fat diets; however, the mechanism is unclear. To determine if the glucose, insulin and cortisol response to isocaloric diets varying in fat and NSC availability differ in fit vs. unfit Thoroughbreds with RER. Four fit (10 weeks treadmill training) RER Thoroughbred mares were exercised and fed 3 isocaloric (121 MJ/day) diets in a 5 day/diet block design. Two high NSC concentrates, sweet feed (SF) and a processed pelleted feed (PL) and a low starch high fat feed (FAT) were used. After 24 h of rest and a 12 h fast, horses ate half their daily concentrate. Blood sampled for [glucose], [insulin] and [cortisol] was obtained before, immediately after and at 30-60 min intervals for 420 min. After 3-6 months detraining period, the block design was repeated. Results for SF and PL were similar. Regardless of diet, cortisol was higher in fit vs. unfit horses. Fit horses on SF/PL had higher post prandial [insulin] and insulin:glucose ratio than unfit horses. FAT resulted in lower post prandial [glucose] and [insulin] vs. SF/PL. Higher [insulin] in fit vs. unfit horses was not seen on the FAT diet. Increased post prandial [glucose], [insulin] and [cortisol] induced by high NSC, but not high fat, feeds are enhanced by fitness in RER horses. This combination may trigger rhabdomyolysis through increased excitability in RER Thoroughbreds.


'Judging a book by its cover': An experimental study of the negative impact of a diagnosis of borderline personality disorder on clinicians' judgements of uncomplicated panic disorder.
Diagnosis is ubiquitous in Psychiatry, and whilst it does bring benefits; adverse effects of 'labelling' may also be possible. This study aimed to evaluate experimentally whether clinicians' judgements about a patient with panic disorder were influenced by an inappropriately suggested diagnosis of comorbid borderline personality disorder (BPD). An experimental design was used to evaluate clinician's judgements when the nature of the information they were given was varied to imply BPD comorbidity. Two hundred and sixty-five clinicians watched a video-recorded assessment of a woman describing her experience of uncomplicated 'panic disorder' and then rated her present problems and likely prognosis. Prior to watching the video recording, participants were randomly allocated to one of three conditions with written information including the following: (1) her personal details and general background; (2) the addition of a behavioural description consistent with BPD; and (3) the further addition of a 'label' (past BPD diagnosis). The BPD label was associated with more negative ratings of the woman's problems and her prognosis than both information alone and a behavioural description of BPD 'symptoms'. Regardless of potential actuarial value of such diagnoses, it is concluded that clinicians can be overly influenced by past diagnostic labels in the context of an apparent current comorbid problem, although such biases appear to be less likely if a description of the relevant behaviours is used instead. Thus, the label, rather than the behaviour it denotes, may be stigmatizing in mental health professionals. Diagnostic labels can have an inappropriately negative effect on clinicians' judgements not only of treatment variables such as engagement and response but also risk issues and interpersonal effectiveness. Diagnostic labels can have a greater effect on clinicians' judgements than a behavioural description or clinical presentation. Clinicians should therefore be cautious both in the use of diagnostic labels to describe patients and ensure that these are still valid, and also be mindful of the influence that such labels can have on their own clinical judgements and constantly seek to challenge these. Behavioural descriptions of difficulties are less likely to result in such negative judgements and predictions.


Ethnic Inequality.
This study explores the consequences and origins of between-ethnicity economic inequality across countries. First, combining satellite images of nighttime luminosity with the historical homelands of ethnolinguistic groups we construct measures of ethnic inequality for a large sample of countries. We also compile proxies of overall spatial inequality and regional inequality across administrative units. Second, we uncover a strong negative association between ethnic inequality and contemporary comparative development; the correlation is also present when we condition on regional inequality, which is itself related to under-development. Third, we investigate the roots of ethnic inequality and establish that differences in geographic endowments across ethnic homelands explain a sizable fraction of the observed variation in economic disparities across groups. Fourth, we show that ethnic-specific inequality in geographic endowments is also linked to under-development.


Crystal structures of the complexes of a group IIA phospholipase A2 with two natural anti-inflammatory agents, anisic acid, and atropine reveal a similar mode of binding.
Secretory low molecular weight phospholipase A(2)s (PLA(2)s) are believed to be involved in the release of arachidonic acid, a precursor for the biosynthesis of pro-inflammatory eicosanoids. Therefore, the specific inhibitors of these enzymes may act as potent anti-inflammatory agents. Similarly, the compounds with known anti-inflammatory properties should act as specific inhibitors. Two plant compounds, (a) anisic acid (4-methoxy benzoic acid) and (b) atropine (8-methyl-8-azabicyclo oct-3-hydroxy-2-phenylpropanoate), have been used in various inflammatory disorders. Both compounds (a) and (b) have been found to inhibit PLA(2) activity having binding constants of 4.5 x 10(-5) M and 2.1 x 10(-8) M, respectively. A group IIA PLA(2) was isolated and purified from the venom of Daboia russelli pulchella (DRP) and its complexes were made with anisic acid and atropine. The crystal structures of the two complexes (i) and (ii) of PLA(2) with compounds (a) and (b) have been determined at 1.3 and 1.2 A resolutions, respectively. The high-quality observed electron densities for the two compounds allowed the accurate determinations of their atomic positions. The structures revealed that these compounds bound to the enzyme at the substrate - binding cleft and their positions were stabilized by networks of hydrogen bonds and hydrophobic interactions. The most characteristic interactions involving Asp 49 and His 48 were clearly observed in both complexes, although the residues that formed hydrophobic interactions with these compounds were not identical because their positions did not exactly superimpose in the large substrate-binding hydrophobic channel. Owing to a relatively small size, the structure of anisic acid did not alter upon binding to PLA(2), while that of atropine changed significantly when compared with its native crystal structure. The conformation of the protein also did not show notable changes upon the bindings of these ligands. The mode of binding of anisic acid to the present group II PLA(2) is almost identical to its binding with bovine pancreatic PLA(2) of group I. On the other hand, the binding of atropine to PLA(2) is similar to that of another plant alkaloid aristolochic acid.


Discovery of Bioactive Natural Products for the Treatment of Acute Respiratory Infections - An Integrated Approach.
In this work, an integrated approach for the identification of new antiviral agents from natural sources for the treatment of acute respiratory infections is presented. The approach comprises (i) the selection of starting material based on traditional knowledge, (ii) phenotypic screening of extracts for antiviral activity, and (iii) the implementation of in silico predictions to identify antiviral compounds and derive the molecular mechanism underlying their biological activity. A variety of starting materials from plants and fungi was selected for the production of 162 extracts. These extracts were tested in cytopathic effect inhibition assays against influenza virus A/Hong Kong/68 (HK/68), rhinovirus A2 (RV-A2), and coxsackie virus B3 (CV-B3). All extracts were also evaluated regarding their cytotoxicity. At an IC50 threshold of 50 µg/mL, 20, 11, and 14% of all tested extracts showed antiviral activity against HK/68, CV-B3, and RV-A2, respectively. Among all active extracts (n = 47), 68% showed antiviral activity against one of the investigated viruses, whereas 31% inhibited at least two viruses. Herein, we present a comprehensive dataset of probed extracts along with their antiviral activities and cytotoxicity. Application examples presented in this work illustrate the phytochemical workflow for the identification of antiviral natural compounds. We also discuss the challenges, pitfalls, and advantages of the integrated approach.


Toxoplasma gondii as a possible causative pathogen of type-1 diabetes mellitus: Evidence from case-control and experimental studies.
Toxoplasma gondii is the causative parasite of an important worldwide disease. This obligate intracellular parasite can infect and replicate inside any nucleated cells including those of pancreas. Insulin is a hormone secreted by the pancreas and is responsible for controlling blood glucose concentration. Deficiency of insulin production accounts for the occurrence of type-1 diabetes mellitus (T1D). Thus, theoretically, toxoplasmosis could play a possible role in the development of T1D. However, the studies on this theory are still insufficient; therefore, this work was designed. Interestingly, in the case-control study, seropositivity of anti-Toxoplasma IgG was significantly higher among T1D (86.37%) in comparison with T2D (66.67%) and the control group (60%). Moreover, the odd ratio of chronic toxoplasmosis was 4.2 folds higher among T1D patients than among controls. The experimental study included acute and chronic Me49 T. gondii infected mice groups in addition to a control group. Pathological examination revealed the presence of T. gondii zoites adjacent to the islets of Langerhans and in pancreatic parenchyma of acutely infected mice. With chronic infection, there was a significant reduction of islets number and sizes in association with grade-1 insulitis. Additionally, the immunohistochemical study showed significant infiltration of the islets of chronically infected mice by CD8+ and CD45+ immune cells. In contrary to the control group, the islets of the chronic group showed significantly higher expression of the apoptotic marker caspase-3 and a significantly lower expression of the proliferation marker Ki69. Finally, a significant reduction of insulin expression in the islets of chronic infection group was detected in association with a significant increase in serum glucose concentrations; however, the establishment of diabetes did not occur throughout this work. Thus, this study presents an evidence for the probable role of chronic toxoplasmosis in the development of T1D which should be considered in further studies.


The single nucleotide polymorphism site of aquaporin-4 gene in patients with neuromyelitis optica.
The single nucleotide polymorphism (SNP) site within the aquaporin (AQP)-4 gene exons and its possible role in the pathogenesis of neuromyelitis optica (NMO) were studied. From March 2010 to June 2012, 72 patients with NMO from Xiangyang No. 1 People's Hospital, Hubei University of Medicine were enrolled in the NMO group. At the same time, 80 patients with multiple sclerosis (MS) were enrolled in our study as the MS group. Blood samples were collected and DNA was extracted for analysis of SNP sites of AQP4 gene. Specific site-directed mutagenesis method was used for site-directed mutagenesis on plasmid enhanced green fluorescence protein carrying AQP4 gene. Mutant plasmids were constructed and used for transfecting cell lines. The differences of anti-AQP4 antibody level in the cell line were analyzed. The possible correlation between AQP4 gene SNP sites and the pathogenesis of NMO were analyzed. In the NMO group, 6 SNP sites in AQP4 gene were located in exons 2 and 5. These included R108T, I110N, E280R, D281R, P295R and E317M. There was no SNP site in exons 1, 3 and 4. In the MS group, no SNP site was found in AQP4 gene. R108T, I110N, R108T/I110N, E280R/D281R, P295R and E317M cell lines were constructed in the NMO group, and anti-AQP4 antibody in the serum was compared between R108T/I110N, E280R/D281R and E317M cell lines and the original HEK293T cell line. The difference was statistically significant (P<0.05). The positive rate of anti-AQP4 antibody titer in serum was compared between R108T, I110N, R108T/I110N, E280R/D281R, P295R and E317M cell lines in the NMO group and the original cell line in the MS group. In conclusion, SNP sites in AQP4 gene in patients with NMO may lead to some conformational changes in AQP4 protein. This affects the antigenicity of AQP4 protein. The different intensity of antigen-antibody reaction may cause the differences of titer observed between the different mutant cell lines.