Construction of a full-length murine pro alpha 2(I) collagen cDNA by the polymerase chain reaction.
Construction of large collagen cDNA has been hindered by the relatively large size and high G-C content of processed mRNA. We describe here the development of a rapid and efficient method for obtaining large full-length collagen cDNA. A full-length (4.3 kb) murine pro alpha 2(I) collagen cDNA was constructed by synthesis of a first-strand cDNA library with use of poly-A RNA (MC3T3-E1) and the oligo-dT17-adapter primer described by Frohman et al (Proc Natl Acad Sci USA 85:8998, 1988). Pro alpha 2(I) collagen cDNA were specifically amplified by the polymerase chain reaction (PCR) with a pro alpha 2(I) specific primer as the 5' primer (20mer; corresponding to nucleotide positions 42-61 in the first exon of the murine pro alpha 2(I) collagen gene, COL1A2), and with the adapter sequence 5' to the dT17 as the 3' primer. The PCR conditions were optimized to allow amplification of the expected 4.0-5.0-kb product; a major 4.3-kb product was visualized by ethidium bromide, identified by in situ gel hybridization, and cloned. DNA sequencing determined that it contained the correct 5' sequence and the 3' end had a 68 basepair (bp) 3' untranslated region. The entire sequence that codes the amino-terminal propeptide domain has been determined and compared to the human sequence. The homology between human and mouse is less in the amino terminal propeptide than in the triple helical domain; exon 5 of murine COL1A2 codes for an additional six amino acids not found in human COL1A2.


Urinary metabolomics fingerprinting around parturition identifies metabolites that differentiate lame dairy cows from healthy ones.
Lameness is a very important disorder of periparturient dairy cows with implications on milk production and composition as well as with consequences on reproductive performance. The aetiology of lameness is not clear although there have been various hypotheses suggested over the years. The objective of this study was to metabotype the urine of dairy cows prior to, during and after the onset of lameness by evaluating at weeks -8, -4 pre-calving, the week of lameness diagnosis, and +4 and +8 weeks post-calving. We used a metabolomics approach to analyse urine samples collected from dairy cows around calving (6 cows with lameness v. 20 healthy control cows). A total of 153 metabolites were identified and quantified using an in-house MS library and classified into 6 groups including: 11 amino acids (AAs), 39 acylcarnitines (ACs), 3 biogenic amines (BAs), 84 glycerophospholipids, 15 sphingolipids and hexose. A total of 23, 36, 40, 23 and 49 metabolites were observed to be significantly different between the lame and healthy cows at -8 and -4 weeks pre-calving, week of lameness diagnosis as well as at +4 and +8 weeks post-calving, respectively. It should be noted that most of the identified metabolites were elevated; however, a few of them were also lower in lame cows. Overall, ACs and glycerophospholipids, specifically phosphatidylcholines (PCs), were the metabolite groups displaying the strongest differences in the urine of pre-lame and lame cows. Lysophosphatidylcholines (LysoPCs), although to a lesser extent than PCs, were altered at all time points. Alterations in urinary AA concentrations were also observed during the current study for four time points. During the pre-calving period, there was an observed elevation of arginine (-8 week), tyrosine (-8 week) and aspartate (-4 week), as well as a depression of urinary glutamate (-4 weeks). In the current study, it was additionally observed that concentrations of several sphingomyelins and one BA were altered in pre-lame and lame cows. Symmetric dimethylarginine was elevated at both -8 weeks pre-calving and the week of lameness diagnosis. Data showed that urinary fingerprinting might be a reliable methodology to be used in the future to differentiate lame cows from healthy ones.


A new DNA marker, U15557, is linked to protease inhibitor and adenylate kinase-1 in the laboratory opossum, Monodelphis domestica.
A 323-bp DNA fragment (U15557) was isolated, cloned, and sequenced after polymerase chain reaction (PCR) amplification from Monodelphis domestica genomic DNA. A HindIII restriction fragment length polymorphism was identified in this species using the U15557 PCR, fragment as a hybridization probe. DNA samples exhibited either a 6.4 kb band, a 7.2 kb band, or both bands simultaneously. Behaviour of these two variants in family studies was consistent with codominant autosomal inheritance. Linkage between this marker and the loci encoding protease inhibitor (PI) and adenylate kinase 1 (AK1) was found in M. domestica.


Loss of heterozygosity at 18q21 region in gastric cancer involves a number of cancer-related genes and correlates with stage and histology, but lacks independent prognostic value.
Several studies support a role of 18q21 LOH, involving the DCC locus, in colorectal cancer progression; however, its contribution to the natural history of gastric cancer is less clear. Recently, a number of cancer-related genes have been mapped in the 18q21 region, either centromeric or telomeric to DCC. This study searched for 18q21 LOH in 161 gastric cancers representative of all tumour stages and main histological types. To this purpose, seven highly polymorphic markers were used flanking the 18q21 band and spanning the entire region. Thirty-four out of 147 (23.1%) informative cases showed LOH. In 27 of 34 cases (79%), LOH involved all the informative loci. The remaining seven cases showed LOH at more telomeric sites and retained heterozygosity at more centromeric markers, mostly those proximal to the DCC gene. A strong correlation between 18q21 LOH and level of gastric wall invasion, lymph node metastases, or stage was found in cohesive (glandular+solid) and mixed tumours, but not in diffuse cancers. Cox univariate and multivariate analysis showed that invasion level, lymph node metastases, distant metastases, TNM stage, and histology were effective predictors of survival, whereas 18q21 LOH did not show predictive power. The simultaneous deletion of a variety of cancer-related genes with different and even opposite roles might explain why, apparently, 18q21 LOH does not per se contribute significantly to the natural history of gastric cancer, despite strong correlation with stage.


A novel method of measuring the melting point of animal fats.
The melting point (TM) of fat is relevant to health, but available methods of determining TM are cumbersome. One of the standard methods of measuring TM for animal and vegetable fats is the slip point, also known as the open capillary method. This method is imprecise and not amenable to automation or mass testing. We have developed a technique for measuring TM of animal fat using the Rotor-Gene Q (Qiagen, Hilden, Germany). The assay has an intra-assay SD of 0.08°C. A single operator can extract and assay up to 250 samples of animal fat in 24 h, including the time to extract the fat from the adipose tissue. This technique will improve the quality of research into genetic and environmental contributions to fat composition of meat.


Re-reading Discourse and Social Psychology: transforming social psychology.
This paper considers one theme in the contemporary legacy of Potter and Wetherell's (1987) Discourse and Social Psychology. It overviews the context that led to that book and considers a series of critical responses from both experimental and critical/qualitative social psychologists. It refutes criticisms and corrects confusions. Focusing on contemporary discursive psychology, it highlights (a) its rigorous use of records of actual behaviour; (b) its systematic focus on normative practices. In methodological terms, it (a) highlights limitations in the use of open-ended interviews; (b) considers the way naturalistic materials provide access to participants' own orientations and displays; (c) builds a distinctive logic of sampling and generalization. In theoretical terms, it (a) highlights the way discourse work can identify foundational psychological matters; (b) offers a novel approach to emotion and embodiment; (c) starts to build a matrix of dimensions which are central to the constructing and recognizing of different kinds of social actions. It now offers a fully formed alternative social psychology which coordinates theory and method and a growing body of empirical work.


Effects of time of feeding during gestation on sow's performance1.
The aim of this study was to investigate the effect of different feeding time regimes given similar energy intake per kilogram live BW0.75 during gestation on sow's performance. One hundred and seventy-four sows [Topigs TN 70 (Landrace × Large White, Topigs USA); parity 3.81 ± 0.16; initial BW = 211.57 ± 3.34 kg; backfat (BF) 13.70 ± 0.42 mm] were blocked by parity, farrowing date, balanced for BW and randomly assigned to 1 of 3 treatments in a randomized complete block design. Treatments included sows fed corn-soybean meal-based diet once at [0730 (control, T1), 1130 (T2), or 1530 h (T3)], with daily feed quantity kept at 1.25× maintenance energy intake [100 × (BW)0.75] kcal ME/d. Sows received 6,758, 7,434, and 8,110 kcal ME/d from days 30 to 60, days 61 to 90, days 91 to 109 of gestation, respectively. The gestation diet was formulated to contain 3,379 kcal of ME/kg, 0.70% Ca, 0.61% total P, 0.58% SID Lys, 0.26% SID Met, 0.45% SID Thr, 0.12% SID Trp, and 0.48% SID Met+Cys. Body weight and BF were recorded on days 30, 60, 90, and 109 of gestation, 24 h after farrowing and at weaning. Results showed that feeding times evaluated did not alter BW changes from day 30 to day 109 of gestation (P = 0.81) or from day 30 to weaning (P = 0.87). Similarly, feeding sows daily at 1130 h did not influence BF gains and sow reproductive performance relative to the control sows (P > 0.10). Sows fed once daily at 1530 h gained more BF compared with the control (3.69 ± 0.47 vs. 2.12 ± 0.50 mm, P = 0.04) from day 30 to day 109 of gestation. From day 30 of gestation to weaning, treatments did not influence BF gain (P = 0.24). Feeding sows daily meal at 1530 h had propensity to increase (P = 0.09) the number of piglets weaned by 0.54 piglets compared with the control sows. In conclusion, the present study demonstrated that feeding pregnant sows at 1530 h altered energy and nutrient metabolism improving their BF gain and exhibited a potential to increase the number of weaned piglets compared with conventional feeding regime.


Pharmacokinetics of primaquine in man. II. Comparison of acute vs chronic dosage in Thai subjects.
We have investigated the pharmacokinetics of primaquine after acute and chronic administration of the drug to five healthy Thai volunteers. After acute dosage (15 mg p.o.) mean (+/- s.d.) peak plasma concentrations of 65.0 +/- 34.7 ng ml-1 were achieved within 2 +/- 1h. Thereafter plasma drug concentrations declined monoexponentially with a mean elimination half life of 4.4 +/- 1.4 h. The mean (+/- s.d.) oral clearance was 37.6 +/- 15.5 1 h-1. These values are in broad agreement with values obtained in healthy Caucasians after administration of an equivalent dose of primaquine. Repeated dosing with primaquine had no effect on the mean pharmacokinetic parameters calculated for this drug. In contrast, individual pharmacokinetic parameters for some subjects exhibited gross and unpredictable changes after chronic dosage. The carboxylic acid metabolite of primaquine accumulated in plasma after repeated dosing such that by day 14 of chronic dosing the mean AUC (0,24) for this metabolite was 74% greater than that obtained after acute administration of primaquine.


Single Marker and Haplotype-Based Association Analysis of Semolina and Pasta Colour in Elite Durum Wheat Breeding Lines Using a High-Density Consensus Map.
Association mapping is usually performed by testing the correlation between a single marker and phenotypes. However, because patterns of variation within genomes are inherited as blocks, clustering markers into haplotypes for genome-wide scans could be a worthwhile approach to improve statistical power to detect associations. The availability of high-density molecular data allows the possibility to assess the potential of both approaches to identify marker-trait associations in durum wheat. In the present study, we used single marker- and haplotype-based approaches to identify loci associated with semolina and pasta colour in durum wheat, the main objective being to evaluate the potential benefits of haplotype-based analysis for identifying quantitative trait loci. One hundred sixty-nine durum lines were genotyped using the Illumina 90K Infinium iSelect assay, and 12,234 polymorphic single nucleotide polymorphism (SNP) markers were generated and used to assess the population structure and the linkage disequilibrium (LD) patterns. A total of 8,581 SNPs previously localized to a high-density consensus map were clustered into 406 haplotype blocks based on the average LD distance of 5.3 cM. Combining multiple SNPs into haplotype blocks increased the average polymorphism information content (PIC) from 0.27 per SNP to 0.50 per haplotype. The haplotype-based analysis identified 12 loci associated with grain pigment colour traits, including the five loci identified by the single marker-based analysis. Furthermore, the haplotype-based analysis resulted in an increase of the phenotypic variance explained (50.4% on average) and the allelic effect (33.7% on average) when compared to single marker analysis. The presence of multiple allelic combinations within each haplotype locus offers potential for screening the most favorable haplotype series and may facilitate marker-assisted selection of grain pigment colour in durum wheat. These results suggest a benefit of haplotype-based analysis over single marker analysis to detect loci associated with colour traits in durum wheat.


Radiotherapy in early-stage Dupuytren's contracture. Long-term results after 13 years.
In early-stage Dupuytren's contracture, radiotherapy is applied to prevent disease progression. Long-term outcome and late toxicity of the treatment were evaluated in a retrospective analysis. Between 12/1982 and 02/2006, 135 patients (208 hands) were irradiated with orthovoltage (120 kV; 20 mA; 4-mm Al filter), in two courses with five daily fractions of 3.0 Gy to a total dose of 30 Gy; separated by a 6- to 8-week interval. The extent of disease was described according to a modified classification of Tubiana et al. Long-term outcome was analyzed at last follow-up between 02/2008 and 05/2008 with a median follow-up of 13 years (range, 2-25 years). Late treatment toxicity and objective reduction of symptoms as change in stage and numbers of nodules and cords were evaluated and used as evidence to assess treatment response. According to the individual stages, 123 cases (59%) remained stable, 20 (10%) improved, and 65 (31%) progressed. In stage N 87% and in stage N/I 70% remained stable or even regressed. In more advanced stages, the rate of disease progression increased to 62% (stage I) or 86% (stage II). 66% of the patients showed a long-term relief of symptoms (i.e., burning sensations, itching and scratching, pressure and tension). Radiotherapy did not increase the complication rate after surgery in case of disease progression and only minor late toxicity (skin atrophy, dry desquamation) could be observed in 32% of the patients. There was no evidence for a second malignancy induced by radiotherapy. After a mean follow-up of 13 years radiotherapy is effective in prevention of disease progression and improves patients' symptoms in early-stage Dupuytren's contracture (stage N, N/I). In case of disease progression after radiotherapy, a "salvage" operation is still feasible.


Shape-based computer-aided detection of lung nodules in thoracic CT images.
In this paper, a new computer tomography (CT) lung nodule computer-aided detection (CAD) method is proposed for detecting both solid nodules and ground-glass opacity (GGO) nodules (part solid and nonsolid). This method consists of several steps. First, the lung region is segmented from the CT data using a fuzzy thresholding method. Then, the volumetric shape index map, which is based on local Gaussian and mean curvatures, and the "dot" map, which is based on the eigenvalues of a Hessian matrix, are calculated for each voxel within the lungs to enhance objects of a specific shape with high spherical elements (such as nodule objects). The combination of the shape index (local shape information) and "dot" features (local intensity dispersion information) provides a good structure descriptor for the initial nodule candidates generation. Antigeometric diffusion, which diffuses across the image edges, is used as a preprocessing step. The smoothness of image edges enables the accurate calculation of voxel-based geometric features. Adaptive thresholding and modified expectation-maximization methods are employed to segment potential nodule objects. Rule-based filtering is first used to remove easily dismissible nonnodule objects. This is followed by a weighted support vector machine (SVM) classification to further reduce the number of false positive (FP) objects. The proposed method has been trained and validated on a clinical dataset of 108 thoracic CT scans using a wide range of tube dose levels that contain 220 nodules (185 solid nodules and 35 GGO nodules) determined by a ground truth reading process. The data were randomly split into training and testing datasets. The experimental results using the independent dataset indicate an average detection rate of 90.2%, with approximately 8.2 FP/scan. Some challenging nodules such as nonspherical nodules and low-contrast part-solid and nonsolid nodules were identified, while most tissues such as blood vessels were excluded. The method's high detection rate, fast computation, and applicability to different imaging conditions and nodule types shows much promise for clinical applications.


Differentiation of Bwamba and Pongola viruses by agar-gel diffusion and immunoelectrophoretic techniques.
The agar-gel diffusion (AGD) and the immunoelectrophoretic (IEP) techniques were employed in the differentiation of virus strains of the Bwamba-Pongola group. These techniques were found to be more specific than either the complement fixation (CF) or the neutralization (N) test in the differentiation of Bwamba and Pongola viurses. Of 22 virus strains isolated in Nigeria and previously typed as strains of Bwamba virus by CF and N tests, 12 strains of mosquito origin were definitely classified as Pongola viruses and 10 of human origin as Bwamba viruses by the AGD and IEP techniques.


Building mountain biodiversity: Geological and evolutionary processes.
Mountain regions are unusually biodiverse, with rich aggregations of small-ranged species that form centers of endemism. Mountains play an array of roles for Earth's biodiversity and affect neighboring lowlands through biotic interchange, changes in regional climate, and nutrient runoff. The high biodiversity of certain mountains reflects the interplay of multiple evolutionary mechanisms: enhanced speciation rates with distinct opportunities for coexistence and persistence of lineages, shaped by long-term climatic changes interacting with topographically dynamic landscapes. High diversity in most tropical mountains is tightly linked to bedrock geology-notably, areas comprising mafic and ultramafic lithologies, rock types rich in magnesium and poor in phosphate that present special requirements for plant physiology. Mountain biodiversity bears the signature of deep-time evolutionary and ecological processes, a history well worth preserving.


The global inventor gap: distribution and equality of world-wide inventive effort, 1990-2010.
Applying distance-to-frontier analysis, we have used 2.9 million patents and population data to assess whether the relative capacity of world countries and major regions to create new knowledge and technology has become globally more equal or less equal between 1990 and 2010. We show with the Gini coefficient that the global distribution of inventors has become more equal between major countries and regions. However, this trend has been largely due to the improved performance of only two major countries, China and India. The worst performing regions, totalling a population of almost 2 billion, are actually falling behind. Our results suggest that substantial parts of the global population have fallen further behind countries at the global frontier in their ability to create new knowledge and inventions, and that the catch-up among the least developed and middle-income countries is highly uneven, prompting questions about the nature and future of the global knowledge economy.


Restored and remnant Banksia woodlands elicit different foraging behavior in avian pollinators.
Pollinators and the pollination services they provide are critical for seed set and self-sustainability of most flowering plants. Despite this, pollinators are rarely assessed in restored plant communities, where their services are largely assumed to re-establish. Bird-pollinator richness, foraging, and interaction behavior were compared between natural and restored Banksia woodland sites in Western Australia to assess their re-establishment in restored sites. These parameters were measured for natural communities of varying size and degree of fragmentation, and restored plant communities of high and low complexity for three years, in the summer and winter flowering of Banksia attenuata and B. menziesii, respectively. Bird visitor communities varied in composition, richness, foraging movement distances, and aggression among sites. Bird richness and abundance were lowest in fragmented remnants. Differences in the composition were associated with the size and degree of fragmentation in natural sites, but this did not differ between seasons. Restored sites and their adjacent natural sites had similar species composition, suggesting proximity supports pollinator re-establishment. Pollinator foraging movements were influenced by the territorial behavior of different species. Using a network analysis approach, we found foraging behavior varied, with more frequent aggressive chases observed in restored sites, resulting in more movements out of the survey areas, than observed in natural sites. Aggressors were larger-bodied Western Wattlebirds (Anthochaera chrysoptera) and New Holland Honeyeaters (Phylidonyris novaehollandiae) that dominated nectar resources, particularly in winter. Restored sites had re-established pollination services, albeit with clear differences, as the degree of variability in the composition and behavior of bird pollinators for Banksias in the natural sites created a broad completion target against which restored sites were assessed. The abundance, diversity, and behavior of pollinator services to remnant and restored Banksia woodland sites were impacted by the size and degree of fragmentation, which in turn influenced bird-pollinator composition, and were further influenced by seasonal changes between summer and winter. Consideration of the spatial and temporal landscape context of restored sites, along with plant community diversity, is needed to ensure the maintenance of the effective movement of pollinators between natural remnant woodlands and restored sites.


Transgenic Clustered Regularly Interspaced Short Palindromic Repeat/Cas9-Mediated Viral Gene Targeting for Antiviral Therapy of Bombyx mori Nucleopolyhedrovirus.
We developed a novel antiviral strategy by combining transposon-based transgenesis and the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) system for the direct cleavage of Bombyx mori nucleopolyhedrovirus (BmNPV) genome DNA to promote virus clearance in silkworms. We demonstrate that transgenic silkworms constitutively expressing Cas9 and guide RNAs targeting the BmNPV immediate early-1 (ie-1) and me53 genes effectively induce target-specific cleavage and subsequent mutagenesis, especially large (∼7-kbp) segment deletions in BmNPV genomes, and thus exhibit robust suppression of BmNPV proliferation. Transgenic animals exhibited higher and inheritable resistance to BmNPV infection than wild-type animals. Our approach will not only contribute to modern sericulture but also shed light on future antiviral therapy.IMPORTANCE Pathogen genome targeting has shown its potential in antiviral research. However, transgenic CRISPR/Cas9 system-mediated viral genome targeting has not been reported as an antiviral strategy in a natural animal host of a virus. Our data provide an effective approach against BmNPV infection in a real-world biological system and demonstrate the potential of transgenic CRISPR/Cas9 systems in antiviral research in other species.


Aetiology of schizophrenia: an evolutionary theory.
It is proposed that schizophrenia is a purely genetic condition that arises as a component of the variation generated in the speciation of Homo sapiens through the evolution of the faculty of language. A critical genetic change on the X and Y chromosomes allowed the two hemispheres to develop with a degree of independence. Thus Man's particular capacity for communication has evolved by a process of increasing hemispheric specialization on the basis of persisting diversity for the asymmetry ('right-shift)' factor. Sex differences in age of onset of psychosis, premorbid precursors and outcome are accounted for in terms of a mean difference between the sexes in cerebral asymmetry. The ultimate cause of this difference is the mechanism of sexual selection acting to retain different ranges of variation of the gene on the X and Y chromosomes. These ranges are expressed in terms of variation in hemispheric specialization and psychological function, a boundary of which represents the predisposition of psychosis. The hypothesis is developmental in the sense that the relevant gene has an influence in development and affects a critical aspect of higher nervous system function. But the key evolutionary process of selection of gene variants occurs late, by the mechanism of mate choice.


Ciprofloxacin-Loaded Polyvinylpyrrolidone Foils for the Topical Treatment of Wound Infections with Methicillin-Resistant Staphylococcus aureus (MRSA).
Bacterial infections are a constant challenge in the management of acute and chronic wounds. Chronic wounds, such as diabetic foot ulcers, have increased significantly in the last few years due to the rise of an aging population. A better understanding of the infectious pathophysiological mechanisms is urgently needed along with new options for the treatment of wound infections and wound-healing disorders. New advances in the preparation of biocompatible dressing materials that can be loaded with antimicrobial drugs may improve the topical treatment of infected wounds. In this study, we investigated the antimicrobial activity of polyvinylpyrrolidone (PVP) foils loaded with ciprofloxacin (Cipro-foils) in the presence of acetic acid as a co-solvent. We used ex vivo human wounds that were infected with two bacterial strains: methicillin-resistant Staphylococcus aureus (MRSA) or Pseudomonas aeruginosa (PAO1). The effectiveness of the treatment was demonstrated by the quantification of the living bacteria extracted from the wound and the detection of released immunological mediators in skin extracts and in the skin culture media. We found that Cipro-foils effectively treated the infection with both PAO1 and MRSA. Other than PAO1, MRSA had no lytic activity toward skin proteins. MRSA infections increased cytokines' expression and release. Interestingly, treatment with Cipro-foils could partially counteract these effects.


Pancreatic beta-cell K(ATP) channel activity and membrane-binding studies with nateglinide: A comparison with sulfonylureas and repaglinide.
Nateglinide (A-4166) is an amino acid derivative with insulinotrophic action in clinical development for treatment of type 2 diabetes. The aim of this study was to determine whether nateglinide's interaction at the K(ATP) channel/sulfonylurea receptor underlies its more rapid onset and shorter duration of action in animal models. Binding studies were carried out with membranes prepared from RIN-m5F cells and HEK-293 cells expressing recombinant human sulfonylurea receptor 1 (SUR1). The relative order for displacement of [(3)H]glibenclamide in competitive binding experiments with RIN-m5F cell membranes was glibenclamide > glimepiride > repaglinide > glipizide > nateglinide > L-nateglinide > tolbutamide. The results with HEK-293/recombinant human SUR1 cells were similar with the exception that glipizide was more potent than repaglinide. Neither nateglinide nor repaglinide had any effect on the dissociation kinetics for [(3)H]glibenclamide, consistent with both compounds competitively binding to the glibenclamide-binding site on SUR1. Finally, the inability to measure [(3)H]nateglinide binding suggests that nateglinide dissociates rapidly from SUR1. Direct interaction of nateglinide with K(ATP) channels in rat pancreatic beta-cells was investigated with the patch-clamp method. The relative potency for inhibition of the K(ATP) channel was repaglinide > glibenclamide > nateglinide. Kinetics of the inhibitory effect on K(ATP) current showed that the onset of inhibition by nateglinide was comparable to glibenclamide but more rapid than that of repaglinide. The time for reversal of channel inhibition by nateglinide was also faster than with glibenclamide and repaglinide. These results suggest that the unique characteristics of nateglinide are largely the result of its interaction at the K(ATP) channel.


Falls among union carpenters.
Falls are a leading cause of morbidity and mortality in the construction trades. We identified a cohort of 16,215 active union carpenters, hours worked, and their workers' compensation claims for a 10-year period. The data on this well-defined cohort were used to describe their work-related falls; to define rates of injury and the associated costs; and to identify high-risk groups. Same level falls occurred at a rate of 1.8/200,000 hours worked; falls from elevations at a rate of 2.3/200,000 hours worked. These injuries resulted in direct payments of 0.30 dollars per hour of work or 2.40 dollars per 8-hr day. Mean costs per fall increased with increasing age. Age was not associated with risk of falls from elevations; younger carpenters had modestly reduced rates of falls from the same level. Rates of falls decreased with increasing time in the union. Carpenters whose usual work involved drywall installation or residential work were at highest risk. Falls are a significant public health risk for carpenters and they are responsible for a significant burden of work-related injury costs. While there is a need for prevention of falls from elevations--through training, enforcement of fall protection regulations, improved safety climate, or engineering changes--there is also the need to prevent falls from lower elevations. Differences in risk likely reflect varying exposures and safety practices in different areas of carpentry, as well as training, experience, and job assignments based on longevity in the union.


Minimally Invasive Management of Posterior Urethral Stricture/Stenosis with DVIU and Mitomycin C Injection.
To demonstrate a technique for minimally invasive endoscopic management of posterior urethral strictures, including those at the bladder neck and vesicourethral anastomosis. Herein, we have included endoscopic video footage from 3 patients with posterior urethral strictures, including 1 at the bladder neck, 1 at the vesicourethral anastomosis, and 1 in the bulbomembranous urethra. In each patient, we perform a direct visualization internal urethrotomy (DVIU) with incisions at the 5 and 7 o'clock positions to widen the urethral lumen, followed by injection of 2 mg mitomycin C (MMC) in a total volume of 5 mL sterile water. Herein, we describe our technique for the endoscopic management of posterior urethral strictures, including those in the prostatic urethra and bladder neck. MMC injection, in conjunction with traditional DVIU, adds minimally to the complexity and length of the procedure but may substantially improve long-term surgical outcomes. Bladder outlet obstruction due to stenosis or stricture of the posterior urethra is a common urologic diagnosis whose etiology can often be traced to prior urethral manipulation or iatrogenic trauma. While Americal Urological Assicuation (AUA) guidelines state that dilation or direct visualization internal urethrotomy (DVIU) should be offered for bulbar strictures measuring less than 2 cm in length, recent evidence suggests that DVIU with or without MMC injection may have utility in the management of bladder neck or vesicourethral anastomotic contractures. We have found that DVIU with subsequent MMC injection is a viable minimally invasive approach for the treatment of posterior urethral strictures. While more data are needed to better understand the long-term success rates of these procedures, this approach should be considered for patients with a bladder outlet obstruction secondary to a short stricture of the posterior urethra, bladder neck, or vesicourethral anastomosis.


Fungi have three tetraspanin families with distinct functions.
Tetraspanins are small membrane proteins that belong to a superfamily encompassing 33 members in human and mouse. These proteins act as organizers of membrane-signalling complexes. So far only two tetraspanin families have been identified in fungi. These are Pls1, which is required for pathogenicity of the plant pathogenic ascomycetes, Magnaporthe grisea, Botrytis cinerea and Colletotrichum lindemuthianum, and Tsp2, whose function is unknown. In this report, we describe a third family of tetraspanins (Tsp3) and a new family of tetraspanin-like proteins (Tpl1) in fungi. We also describe expression of some of these genes in M. grisea and a basidiomycete, Laccaria bicolor, and also their functional analysis in M. grisea. The exhaustive search for tetraspanins in fungal genomes reveals that higher fungi (basidiomycetes and ascomycetes) contain three families of tetraspanins (Pls1, Tsp2 and Tsp3) with different distribution amongst phyla. Pls1 is found in ascomycetes and basidiomycetes, whereas Tsp2 is restricted to basidiomycetes and Tsp3 to ascomycetes. A unique copy of each of PLS1 and TSP3 was found in ascomycetes in contrast to TSP2, which has several paralogs in the basidiomycetes, Coprinus cinereus and Laccaria bicolor. A tetraspanin-like family (Tpl1) was also identified in ascomycetes. Transcriptional analyses in various tissues of L. bicolor and M. grisea showed that PLS1 and TSP2 are expressed in all tissues in L. bicolor and that TSP3 and TPL1 are overexpressed in the sexual fruiting bodies (perithecia) and mycelia of M. grisea, suggesting that these genes are not pseudogenes. Phenotypic analysis of gene replacementmutants Deltatsp3 and Deltatpl1 of M. grisea revealed a reduction of the pathogenicity only on rice, in contrast to Deltapls1 mutants, which are completely non-pathogenic on barley and rice. A new tetraspanin family (Tsp3) and a tetraspanin-like protein family (Tpl1) have been identified in fungi. Functional analysis by gene replacement showed that these proteins, as well as Pls1, are involved in the infection process of the plant pathogenic fungus M. grisea. The next challenge will be to decipher the role(s) of tetraspanins in a range of symbiotic, saprophytic and human pathogenic fungi.


Feasibility and Clinical Effectiveness of Three-Dimensional Printed Model-Assisted Nuss Procedure.
The Nuss procedure is a minimally invasive technique for correcting pectus excavatum. We hypothesized that three-dimensional (3D) simulation may shorten operation time and provide better morphologic outcome. This study aimed to demonstrate the feasibility of the 3D model-assisted Nuss procedure and to compare its potential benefits with those of the traditional Nuss procedure. We simulated the targeted curvature, length, and planned intercostal space of a metallic bar, based on the preoperative chest computed tomographic images. After the use of a 3D printing technique, a plastic template bar was produced and sterilized. The metallic bar was bent and placed at the planned intercostal space accordingly. The patients' characteristics, total number of pectus bar placement, total operation time, and improvement percentage of Haller indices were compared with patients who underwent the traditional Nuss procedure. A total of 419 patients underwent the Nuss procedure from January 2010 to July 2017 in our hospital, and 357 patients were eligible and enrolled for the following analysis. Fifteen patients underwent 3D simulation. After performing propensity-score matching analysis, the 3D printing group had a shorter operative time (60.36 versus 74.34 minutes, p < 0.001), fewer metallic bar placements (1.00 versus 1.36 bars, p < 0.001), and better improvement percentages in the Haller indices (20.34% versus 10.06%, p < 0.001) compared with the traditional Nuss procedure. In this preliminary study, 3D-printed model-assisted Nuss procedure may provide benefits of shorter operative time, fewer metallic bar insertions, and comparable morphologic outcome by preoperative simulation.


Comparison and optimization of hiPSC forebrain cortical differentiation protocols.
Several protocols have been developed for human induced pluripotent stem cell neuronal differentiation. We compare several methods for forebrain cortical neuronal differentiation by assessing cell morphology, immunostaining and gene expression. We evaluate embryoid aggregate vs. monolayer with dual SMAD inhibition differentiation protocols, manual vs. AggreWell aggregate formation, plating substrates, neural progenitor cell (NPC) isolation methods, NPC maintenance and expansion, and astrocyte co-culture. The embryoid aggregate protocol, using a Matrigel substrate, consistently generates a high yield and purity of neurons. NPC isolation by manual selection, enzymatic rosette selection, or FACS all are efficient, but exhibit some differences in resulting cell populations. Expansion of NPCs as neural aggregates yields higher cell purity than expansion in a monolayer. Finally, co-culture of iPSC-derived neurons with astrocytes increases neuronal maturity by day 40. This study directly compares commonly employed methods for neuronal differentiation of iPSCs, and can be used as a resource for choosing between various differentiation protocols.


Atlantoaxial joint jamming as a treatment for atlantoaxial dislocation: a preliminary report. Technical note.
In this report the author describes an alternative technique of atlantoaxial fixation that involves blocking or "jamming" of movements of the atlantoaxial joint. The technique involves forcible impaction of spiked titanium metal spacers and bone grafts within the distracted atlantoaxial facet joints. Between January 2003 and January 2006, four patients underwent this method of fixation at the Department of Neurosurgery at King Edward Memorial Hospital in Mumbai, India. All four patients had posttraumatic mobile and reducible atlantoaxial dislocation. The mean follow-up period was 16 months (range 5-35 months). Successful atlantoaxial stabilization along with ultimate bone fusion was achieved in all patients and was documented on dynamic radiographs. There were no neurological, vascular, or infection-associated complications. The author concludes that the described method of atlantoaxial fixation provides an alternative treatment strategy. "Joint jamming" as a stand-alone method or in combination with other fixation methods may provide firm stabilization in cases of atlantoaxial dislocation.


Prevalence, incidence, and demographics of systemic lupus erythematosus and lupus nephritis from 2000 to 2004 among children in the US Medicaid beneficiary population.
To investigate the nationwide prevalence, incidence, and sociodemographics of systemic lupus erythematosus (SLE) and lupus nephritis among children in the US Medicaid beneficiary population. Children ages 3 years to <18 years with a diagnosis of SLE (defined as ≥3 claims with an International Classification of Diseases, Ninth Revision [ICD-9] code of 710.0 for SLE, each >30 days apart) were identified from the US Medicaid Analytic eXtract database from 2000 to 2004. This database contains all inpatient and outpatient Medicaid claims for 47 US states and the District of Columbia. Lupus nephritis was identified from ≥2 ICD-9 billing codes for glomerulonephritis, proteinuria, or renal failure, each recorded >30 days apart. The prevalence and incidence of SLE and lupus nephritis were calculated among Medicaid-enrolled children overall and within sociodemographic groups. Of the 30,420,597 Medicaid-enrolled children during these years, 2,959 were identified as having SLE. The prevalence of SLE was 9.73 (95% confidence interval [95% CI] 9.38-10.08) per 100,000 Medicaid-enrolled children. Among the children with SLE, 84% were female, 40% were African American, 25% were Hispanic, 21% were White, and 42% resided in the South region of the US. Moreover, of the children with SLE, 1,106 (37%) had lupus nephritis, representing a prevalence of 3.64 (95% CI 3.43-3.86) per 100,000 children. The average annual incidence of SLE was 2.22 cases (95% CI 2.05-2.40) and that of lupus nephritis was 0.72 cases (95% CI 0.63-0.83) per 100,000 Medicaid enrollees per year. The prevalence and incidence rates of SLE and lupus nephritis increased with age, were higher in girls than in boys, and were higher in all non-White racial/ethnic groups. In the current study, the prevalence and incidence rates of SLE among Medicaid-enrolled children in the US are high compared to studies in other populations. In addition, these data represent the first population-based estimates of the prevalence and incidence of lupus nephritis in the US to date.


Differential induction of long-term synaptic facilitation by spaced and massed applications of serotonin at sensory neuron synapses of Aplysia californica.
Serotonin (5HT)-induced facilitation of synaptic transmission from tail sensory neurons (SNs) to motor neurons (MNs) in the marine mollusc Aplysia provides a cellular model of short- and long-term memory for behavioral sensitization of the tail withdrawal reflex. Synaptic facilitation at these synapses occurs in three temporal phases: short-term (STF, lasting minutes), intermediate-term (ITF, lasting more than an hour), and long-term (LTF, lasting >24 hr). STF, ITF, and LTF differ in their induction requirements: A single brief exposure of 5HT induces STF, whereas five applications are required for ITF and LTF. Moreover, STF and LTF can be induced independently. Different forms of memory often show differential sensitivity to the pattern of training trials. To begin to explore this effect at a cellular level, we examined ITF and LTF induced by one of two patterns of 5HT application: a spaced pattern (five 5-min exposures with an interval of 15 min) or a massed pattern (one continuous 25-min application). The spaced and massed patterns both induced ITF; however, spaced 5HT application was significantly more reliable at inducing LTF than was massed application. Thus, whereas induction of ITF and LTF require similar amounts of 5HT, the cellular mechanisms underlying the induction of LTF are more sensitive to the pattern of the induction trials. In the massed group, further analysis revealed a relationship between the expression of ITF and the subsequent expression of LTF, suggesting that these two processes may be mechanistically related.


Anacardic acid mitigates liver fat accumulation and impaired glucose tolerance in mice fed a high-fat and high-sucrose diet.
In this study, we evaluated the effects of anacardic acid (AA), a phenolic lipid found in cashew nuts (Anacardium occidentale), on metabolic disorders related to obesity, fatty liver disease, and diabetes using both in vitro and in vivo models. The application of AA led to a reduction in lipid accumulation in 3T3-L1 cells without observable cytotoxicity. Results from Western blot analysis revealed that AA treatment also led to decreased expression of fatty acid synthase and peroxisome proliferator-activated receptor gamma. In vivo studies were performed to evaluate the effects of AA treatment on fatty liver disease and diabetes. Mice fed a high-fat and high-sucrose diet had significantly higher body and liver weights, and higher levels of liver fat, cholesterol, fasting glucose, and homeostasis model assessment of insulin resistance (HOMA-IR). However, 12 weeks of oral treatment with 500 μg/kg BW AA slowed down lipid accumulation rates in the liver and mitigated insulin resistance in these mice. Thus, AA may reduce lipid levels and have an antidiabetic effect.


Prevention of growing skull fractures: report of 2 cases.
The author describes 2 cases of children with growing skull fractures (GSFs). Surgical exploration of the widened fracture shortly after the head injury failed to reveal a dural tear because the neuroimaging studies (MR images, CT scans, and skull radiographs) had not been accurately interpreted, thereby allowing the development of a GSF at the site of the actual dural injury. In both cases, the dural and bony defect and the leptomeningeal cyst were successfully repaired. To prevent GSFs associated with progressive neurological deficit, seizure, ventricular porencephaly, and encephalomalacia, the author surgically explores wide skull fractures in young children with head injury whose MR images demonstrate brain herniation through the dura mater. The importance of a brief delay in surgical exploration is emphasized to allow cerebral edema to resolve and the patient's condition to become medically stabilized.


Expression of the KCl cotransporter KCC2 parallels neuronal maturation and the emergence of low intracellular chloride.
Fast synaptic inhibition in the adult central nervous system (CNS) is mediated by GABA and glycine. During early development GABA acts as an excitatory neurotransmitter, which is deemed to be important for the maturation of the CNS. During development GABAergic responses undergo a switch from excitatory to inhibitory. This switch is correlated with upregulation of KCC2, the neuronal isoform of the potassium-chloride cotransporter family. KCC2 lowers the intraneuronal chloride concentration below its electrochemical equilibrium. KCC2 activity is thought to depend on phosphorylation by endogenous tyrosine kinases. Here, we analyzed the expression pattern of KCC2 during murine embryonic and postnatal development by in situ hybridization and Western blot analysis. KCC2 expression paralleled neuronal differentiation and preceded the decline of the GABA reversal potential (EGABA) in spinal cord motoneurons and hippocampal pyramidal cells. The adult inhibitory response to GABA was established earlier in the spinal cord than in the hippocampus. Phosphorylated KCC2 protein was already present early in development when the functional GABA switch had not yet occurred. Thus, tyrosine-phosphorylation seems to be less important than the transcriptional upregulation of KCC2.


Fully degeneracy-lifted bow-tie elliptical ring-core multi-mode fiber.
We present a bow-tie elliptical ring-core multi-mode fiber (BT-ERC-MMF) that features an elliptical ring-core structure and two symmetrical bow-tie stress-applying parts (SAPs). This special fiber design breaking geometry and stress symmetry fully separates the two- or four-fold degenerate modes in traditional circular symmetry fiber with strong mode coupling during mode-division multiplexing (MDM) transmission. The designed fiber is able to support 53 fully degeneracy-lifted eigenmodes with minimum effective index difference between adjacent modes larger than 1.59 × 10-4 at 1550 nm, facilitating potential fiber eigenmode multiplexing transmission without using multiple-input multiple-output digital signal processing (MIMO-DSP) technique. The effect of bending on the designed fiber is investigated based on conformal mapping. Broadband performance including effective modal index (neff), effective index difference (Δneff), effective mode area (Aeff), nonlinearity and chromatic dispersion (D) is also comprehensively studied over the whole C band ranging from 1530 to 1565 nm. The designed fiber targets emerging applications in low-crosstalk direct fiber eigenmode-division multiplexing combined with the mature wavelength-division multiplexing (WDM) technique to increase transmission capacity and spectral efficiency.


Synthesis and properties of dodecyl trehaloside detergents for membrane protein studies.
Sugar-based detergents, mostly derived from maltose or glucose, prevail in the extraction, solubilization, stabilization, and crystallization of membrane proteins. Inspired by the broad use of trehalose for protecting biological macromolecules and lipid bilayer structures, we synthesized new trehaloside detergents for potential applications in membrane protein research. We devised an efficient synthesis of four dodecyl trehalosides, each with the 12-carbon alkyl chain attached to different hydroxyl groups of trehalose, thus presenting a structurally diverse but related family of detergents. The detergent physical properties, including solubility, hydrophobicity, critical micelle concentration (CMC), and size of micelles, were evaluated and compared with the most popular maltoside analogue, β-D-dodecyl maltoside (DDM), which varied from each other due to distinct molecular geometries and possible polar group interactions in resulting micelles. Crystals of 2-dodecyl trehaloside (2-DDTre) were also obtained in methanol, and the crystal packing revealed multiple H-bonded interactions among adjacent trehalose groups. The few trehaloside detergents were tested for the solubilization and stabilization of the nociceptin/orphanin FQ peptide receptor (ORL1) and MsbA, which belong to the G-protein coupled receptor (GPCR) and ATP-binding cassette transporter families, respectively. Our results demonstrated the utility of trehaloside detergents as membrane protein solubilization reagents with the optimal detergents being protein dependent. Continuing development and investigations of trehaloside detergents are attractive, given their interesting and unique chemical-physical properties and potential interactions with membrane lipids.


Clinicopathological and survival analyses of primary cutaneous angiosarcoma in an Asian population: prognostic value of the clinical features of skin lesions.
Primary cutaneous angiosarcoma (CA) is a rare but aggressive tumor with a high rate of local recurrence. This study was designed to analyze the clinicopathological features of primary CA and identify factors of cutaneous manifestations associated with the prognosis of angiosarcoma. Medical records of 55 patients with primary CA were retrospectively analyzed to investigate clinical features, survivals, and prognostic factors. Anatomical location of tumor was classified to the scalp, face, and neck, and sites outside the head and neck. Primary CA presented cutaneous nodules (31/55, 47.2%), patches (13/55, 23.6%), and indurated plaques (11/55, 20.0%). Nodular lesion was significantly more common in CA on the scalp compared to CA on sites outside the scalp. Histologically, tumors presenting as nodular lesions on the scalp was predominantly composed of solid sheets of large pleomorphic cells, whereas non-nodular lesions composed of tumor cells between collagen bundles forming irregular vascular spaces. Cutaneous angiosarcoma on the scalp showed a worse prognosis compared to CA on sites outside the scalp. Patients presenting clinical morphology with nodules and multiple skin lesions showed significantly reduced overall survival (OS). In primary CA, location on the scalp, morphology with nodules, and multiplicity of skin lesions significantly affected survival outcomes.


MODELING TEMPORAL GRADIENTS IN REGIONALLY AGGREGATED CALIFORNIA ASTHMA HOSPITALIZATION DATA.
Advances in Geographical Information Systems (GIS) have led to the enormous recent burgeoning of spatial-temporal databases and associated statistical modeling. Here we depart from the rather rich literature in space-time modeling by considering the setting where space is discrete (e.g., aggregated data over regions), but time is continuous. Our major objective in this application is to carry out inference on gradients of a temporal process in our data set of monthly county level asthma hospitalization rates in the state of California, while at the same time accounting for spatial similarities of the temporal process across neighboring counties. Use of continuous time models here allows inference at a finer resolution than at which the data are sampled. Rather than use parametric forms to model time, we opt for a more flexible stochastic process embedded within a dynamic Markov random field framework. Through the matrix-valued covariance function we can ensure that the temporal process realizations are mean square differentiable, and may thus carry out inference on temporal gradients in a posterior predictive fashion. We use this approach to evaluate temporal gradients where we are concerned with temporal changes in the residual and fitted rate curves after accounting for seasonality, spatiotemporal ozone levels and several spatially-resolved important sociodemographic covariates.


Morphological approach to tumours of the testis and paratestis.
Most neoplastic scrotal masses ultimately prove to be germ cell tumours and are recognisable with routine haematoxylin and eosin-stained sections. The differential diagnosis may be focused, even before reviewing histological sections, by knowledge of patient age, medical history, tumour site (testicular vs paratesticular) and gross findings. Some cases may prove to be diagnostically challenging, including rare tumours, a common tumour with an unusual pattern, a metastatic tumour, or a neoplasm with features that mimic another tumour. Several morphological patterns are seen with some frequency and these generate recurring sets of differential diagnostic considerations. These common patterns include testicular tumours with a predominant diffuse arrangement of cells with pale to clear cytoplasm, tumours with a glandular/tubular pattern, tumours with a microcystic pattern and tumours composed of oxyphilic cells. Intratubular proliferations of atypical cells, paratesticular glandular and/or papillary tumours, or tumours with spindle cell morphology can also be challenging to diagnose correctly. In some problematic cases, immunohistochemical staining may be useful to resolve these differential diagnoses.


HTLV type IIIB infection of human thymic epithelial cells: viral expression correlates with the induction of NF-kappa B-binding activity in cells activated by cell adhesion.
Productive infection by the LAV strain has been demonstrated in T cell precursors at different stages of intrathymic development, while viral replication in thymic epithelial cells is still controversial. In this article we show that epithelial cell cultures derived from the medullary component of normal thymus are infectable by HTLV-IIIB virus through cell-free and lymphoid-mediated transmission. Free virus inoculum results in the integration of proviral copies undergoing poor replication, whereas lymphoid-mediated transmission leads to substantial viral expression and the production of viral progeny able to secondary infect lymphoid cells. Interleukin 6 production and phenotype changes (increased expression of MHC class I and ICAM-1) were induced in TE cells by contact with free virus or by adhesion to infected lymphoid cells. By contrast, NF-kappa B-binding activity on the HIV-1 LTR kappa B enhancer element was upregulated only by contact with infected lymphoid cells, but not with virus. The viral replication observed in TE cells after lymphoid-mediated transmission correlates with the upregulation of NF-kappa B-binding activity. Interleukin 6 increased production and phenotype changes and increased NF-kappa B-binding activity were also induced by adhesion to uninfected lymphoid cells, demonstrating that lymphoepithelial cell contacts can activate TE cells. These results demonstrate that thymic epithelial cells are permissive to HIV infection and that viral replication in this cell lineage can be modulated by intracellular signals delivered by adhesive contacts.


Randomized phase II trial of erlotinib versus temozolomide or carmustine in recurrent glioblastoma: EORTC brain tumor group study 26034.
Approximately 50% of glioblastomas (GBMs) are characterized by overexpression of the epidermal growth factor receptor (EGFR) and EGFR gene amplification. In approximately 25% of instances, constitutively activated EGFR mutants are present. These observations make EGFR-inhibiting drugs a logical approach for trials in recurrent GBM. In a randomized, controlled, phase II trial, 110 patients with progressive GBM after prior radiotherapy were randomly assigned to either erlotinib or a control arm that received treatment with either temozolomide or carmustine (BCNU). The primary end point was 6-month progression-free survival (PFS). Tumor specimens obtained at first surgery were investigated for EGFR expression; EGFRvIII mutants; EGFR amplification; EGFR mutations in exons 18, 19, and 21; and pAkt. These results were correlated with outcome. Pharmacokinetic analysis was part of the study. RESULTS; Treatment was well tolerated in general; skin toxicity was the most frequent adverse effect of erlotinib. The 6-month PFS rate in the erlotinib arm was 11.4% (95% CI, 4.6% to 21.5%), and it was 24% in the control arm. Of all explored biomarkers, only low pAkt expression appeared to be of borderline significance to an improved outcome. None of the eight patients who had tumors with EGFRvIII mutant presence and PTEN expression had 6-month PFS. The use of enzyme-inducing anticonvulsants significantly increased erlotinib clearance, but pharmacokinetic findings were not related to outcome. Erlotinib has insufficient single-agent activity in unselected GBM. No clear biomarker associated with improved outcome to erlotinib was identified.


Prediction of the Chemoreflex Gain by Common Clinical Variables in Heart Failure.
Peripheral and central chemoreflex sensitivity, assessed by the hypoxic or hypercapnic ventilatory response (HVR and HCVR, respectively), is enhanced in heart failure (HF) patients, is involved in the pathophysiology of the disease, and is under investigation as a potential therapeutic target. Chemoreflex sensitivity assessment is however demanding and, therefore, not easily applicable in the clinical setting. We aimed at evaluating whether common clinical variables, broadly obtained by routine clinical and instrumental evaluation, could predict increased HVR and HCVR. 191 patients with systolic HF (left ventricular ejection fraction--LVEF--<50%) underwent chemoreflex assessment by rebreathing technique to assess HVR and HCVR. All patients underwent clinical and neurohormonal evaluation, comprising: echocardiogram, cardiopulmonary exercise test (CPET), daytime cardiorespiratory monitoring for breathing pattern evaluation. Regarding HVR, multivariate penalized logistic regression, Bayesian Model Averaging (BMA) logistic regression and random forest analysis identified, as predictors, the presence of periodic breathing and increased slope of the relation between ventilation and carbon dioxide production (VE/VCO2) during exercise. Again, the above-mentioned statistical tools identified as HCVR predictors plasma levels of N-terminal fragment of proBNP and VE/VCO2 slope. In HF patients, the simple assessment of breathing pattern, alongside with ventilatory efficiency during exercise and natriuretic peptides levels identifies a subset of patients presenting with increased chemoreflex sensitivity to either hypoxia or hypercapnia.


Orbital richter syndrome.
We report two patients with previously diagnosed chronic lymphocytic leukemia who developed Richter syndrome in the orbit as the sole extranodal site. The medical history, clinical findings, orbital imaging and histopathological features of the patients were reviewed. Treatment protocols and the outcomes were also assessed. The first patient developed Richter syndrome at the age of 64 years, 3 years after the diagnosis of chronic lymphocytic leukemia. The tumor was located at the inferotemporal quadrant of the orbit. The second patient was 59 years old when Richter syndrome arose in the lacrimal gland, 4 years after the diagnosis of chronic lymphocytic leukemia. Incisional biopsy from the orbital tumors were performed. Histopathological findings included diffuse CD20, CD 23, CD5, bcl2, bcl6 positive lymphocytic infiltration. Both patients were treated with chemotherapy and rituximab. During 3 years of follow-up, there was no orbital or systemic recurrence of the disease. Richter syndrome may develop in the orbital soft tissue and the lacrimal gland, and the orbital disease appears to have a better prognosis compared to patients with systemic involvement.


[Gastric carcinoma in patients under 35 years].
To address the frequency, type of clinical presentation, treatment modalities and survival of gastric carcinoma in young Mexicans. Hospital de Especialidades, Centro Medico Nacional Siglo XXI, IMSS, Mexico City. A retrospective review of all charts of patients 35 years old or younger, with a diagnosis of gastric carcinoma treated at the department of surgery, from July 1986 to January 1990 was performed. Follow up was conducted at the surgery clinic until death or up to January 1998. Eleven patients under 35 years of age were identified, they represent 13.7% of 80 patients treated during that time period. Women were affected more frequently (1.7:1). Ten patients presented with advanced disease, and only one patient had Stage II. Five gastric resections were performed, one patient had a bypass only, three underwent surgical explorations and two were not operated. Ten had diffuse type lesions and only one intestinal adenocarcinoma. Median survival was 15.3 months and only one patient is alive and well. In our series, gastric carcinoma is frequent, it is diagnosed in advanced stages, and this is the reason for a very poor prognosis.


Morphological transformation of established rodent cell lines by high-level expression of the adenovirus type 2 E1a gene.
When a strong promoter derived from the mouse metallothionein gene was substituted for the homologous adenovirus type 2 E1a promoter, leading to enhanced levels of E1a RNAs and proteins in cells transfected with the chimeric gene, the E1a gene alone was able to induce in established cell lines alterations in cellular morphology and growth properties similar to those produced by the combined action of E1a and E1b genes. The qualitative effects of E1a gene expression upon cellular properties thus depend on the level of expression of the E1a gene. Furthermore, E1a may be the primary transforming gene of adenoviruses, since it produced many of the characteristics of transformed cells that had previously been attributed to E1b.


Exploring Research Engagement and Priorities of Transgender and Gender Diverse Veterans.
In recent years, the U.S. Veterans Health Administration (VHA) has seen an increasing population of transgender and gender diverse (TGD) veterans accessing care. Approximately 139 per 100,000 VHA users had a gender identity disorder diagnosis documented in 2018 compared to 32.9 per 100,000 in 2013. Despite TGD patients being overrepresented within VHA, TGD veterans may distrust or face unique barriers with various aspects of the VHA, including health services research. Existing VHA health research focused on TGD populations is largely limited to secondary analyses of electronic health record data. Identifying strategies to enhance primary data collection is crucial for more deeply investigating health care challenges experienced by TGD veterans using VHA care. Additionally, describing health topics of importance for TGD veterans is important for making the research agenda more patient-centered. In this study, we offer veterans' recommendations for researchers working with underrepresented populations based on our findings. From September through October 2019, 30 TGD veterans were recruited through VHA lesbian, gay, bisexual, transgender, and queer/questioning Veteran Care Coordinators (LGBTQ+ VCC) located nationwide. Semi-structured interviews were used to explore barriers and facilitators to research participation, recommendations for improving outreach and engagement, and overall perspectives about priorities in health services research. Transcripts were independently and jointly reviewed and coded by two TGD research analysts, including a veteran using VHA care. Codes were derived inductively. Themes were identified using conventional content analysis. The VA Pittsburgh Healthcare System institutional review board approved this study. Participants cited privacy concerns of being "outed" and potentially having VHA benefits revoked, in addition to a level of distrust in researchers' intentions as barriers to participating in studies. Facilitators for participating included feeling a sense of serving the TGD community and accessibility to study locations, especially VHA-affiliated sites. Suggestions for recruitment included tailored messaging and using other TGD peers or affirming VHA staff (e.g., LGBTQ+ VCCs) for study outreach. Mental health and gender-affirming hormone therapy were the most understudied topics identified by participants. Additionally, participants prioritized the inclusion and study of underrepresented subpopulations, such as transgender women of color, transgender men, and non-binary/gender diverse veterans, in future research. By harnessing the VHA LGBTQ+ VCC network, this study recruited a national sample of TGD veterans to provide insight on methods for more effectively engaging TGD veterans in research and elicited their suggestions for health services research topics. The findings provide numerous suggestions for medicine and public health that are ripe for future research endeavors. Despite the study's lack of gender, racial, and ethnic diversity, findings highlight the need for engagement and study of underrepresented veteran populations. These suggested areas of focus for research in combination with valuable insight on research participation provide researchers with guidance for developing research agendas and designing recruitment and data collection methods that can facilitate future primary research advancing health services research involving TGD patients. Similarly, VHA and non-VHA researchers conducting research involving other underrepresented populations can also gain insight from these findings.


[Selection of cell lines resistant to tryptophan analogs].
After prolonged cultivation in the presence of increasing amounts of carboxyl-substituted tryptophan analogs (tryptamine and tryptophanol), cell lines resistant to high concentrations of these compounds were obtained. The initial culture was the Madin-Darby line of spontaneously transformed bovine kidney cells. In the resistant lines the amount of tryptophanyl-tRNA synthetase (E. C. 18.104.22.168) is manyfold increased as shown by two criteria: (i) enzymatic activity (ATP-PPi isotopic exchange) per mg of protein, (ii) binding of in vivo 35S-labeled proteins to polyclonal antibodies against tryptophanyl-tRNA synthetase. It was shown that tryptophanyl-tRNA synthetase is phosphorylated in vivo, and the degree of phosphorylation of the enzyme in initial cells seems to be higher then in the resistant ones. The Km value for tryptophan is not significantly changed for the enzyme from resistant cells. The permeability for tryptophan and its analogs is reduced in the resistant cells. It is proposed that the acquisition of the resistance against tryptophan analogs are due to alterations at the genomic level (for example, gene amplification etc.).


Root length is proxy for high-throughput screening of waterlogging tolerance in Urochloa spp. grasses.
C4 perennial Urochloa spp. grasses are widely planted in extensive areas in the tropics. These areas are continuously facing waterlogging events, which limits plant growth and production. However, no commercial cultivar combining excellent waterlogging tolerance with superior biomass production and nutritional quality is available. The objective of this study was to identify root traits that can be used for selecting waterlogging tolerant species of Urochloa. Root respiration, root morphological, architectural and anatomical traits were evaluated in eight contrasting Urochloa spp. genotypes grown under aerated or deoxygenated stagnant solutions. Moreover, modelling of internal aeration was used to relate differences in root traits and root growth in waterlogged soils. Increased aerenchyma formation in roots, reduced stele area and development of a fully suberised exodermis are characteristics improving internal aeration of roots and therefore determining waterlogging tolerance in these C4 forage grasses. Waterlogging-tolerant genotypes had steeper root angles and greater root lengths than the waterlogging-sensitive genotypes. In stagnant conditions, waterlogging-tolerant genotypes had a greater proportion of aerenchyma and reduced stele area in root cross-sections, had deeper roots, steeper root angle and larger root biomass, which in turn, allowed for greater shoot biomass. Total root length had the strongest positive influence on shoot dry mass and can therefore be used as proxy for selecting waterlogging tolerant Urochloa genotypes.


Capsaicin, an inhibitor of Ochratoxin A production by Aspergillus section Nigri strains in grapes (Vitis vinifera L.).
Food decay by spoilage fungi leads to significant economic losses and hazards to consumers' health due to the potential of mycotoxin occurrence. Ochratoxin A (OTA) is a mycotoxin known as nephrotoxic and carcinogenic to humans. Natural capsaicin was evaluated for its effectiveness against the growth of five Aspergillus section Nigri strains and accumulation of OTA in inoculated black grapes. Results showed that capsaicin was effective in inhibiting fungal growth and OTA production by new four Aspergillus section Nigri strains (ATHUM 6997, 6998, 6999, 7000) and by Aspergillus carbonarius as well. Moreover, capsaicin addition exhibited maximum inhibition of OTA produced by ATHUM 6997, 6998, 6999, and 7000 in black grapes at 28.9%, 8.6%, 68.4%, and 78.1%, respectively. Inhibition percentage of OTA production by A. carbonarius in grapes treated with capsaicin was estimated at 61.5%. These results suggest that capsaicin influences the OTA biosynthesis pathway of all Aspergillus section Nigri strains and therefore could be used as an effective natural preservative against OTA contamination of vineyards. Risk assessment revealed that when grapes are treated with capsaicin, consumers are less exposed to OTA.


ICTV Virus Taxonomy Profile: Cystoviridae.
The family Cystoviridae includes enveloped viruses with a tri-segmented dsRNA genome and a double-layered protein capsid. The innermost protein shell is a polymerase complex responsible for genome packaging, replication and transcription. Cystoviruses infect Gram-negative bacteria, primarily plant-pathogenic Pseudomonas syringae strains. This is a summary of the International Committee on Taxonomy of Viruses (ICTV) Report on the taxonomy of the Cystoviridae, which is available at http://www.ictv.global/report/cystoviridae.


Neurotrophic effects of progranulin in vivo in reversing motor neuron defects caused by over or under expression of TDP-43 or FUS.
Progranulin (PGRN) is a glycoprotein with multiple roles in normal and disease states. Mutations within the GRN gene cause frontotemporal lobar degeneration (FTLD). The affected neurons display distinctive TAR DNA binding protein 43 (TDP-43) inclusions. How partial loss of PGRN causes TDP-43 neuropathology is poorly understood. TDP-43 inclusions are also found in affected neurons of patients with other neurodegenerative diseases including amyotrophic lateral sclerosis (ALS) and Alzheimer's disease. In ALS, TDP-43 inclusions are typically also immunoreactive for fused in sarcoma (FUS). Mutations within TDP-43 or FUS are themselves neuropathogenic in ALS and some cases of FTLD. We used the outgrowth of caudal primary motor neurons (MNs) in zebrafish embryos to investigate the interaction of PGRN with TDP-43 and FUS in vivo. As reported previously, depletion of zebrafish PGRN-A (zfPGRN-A) is associated with truncated primary MNs and impaired motor function. Here we found that depletion of zfPGRN-A results in primary MNs outgrowth stalling at the horizontal myoseptum, a line of demarcation separating the myotome into dorsal and ventral compartments that is where the final destination of primary motor is assigned. Successful axonal outgrowth beyond the horizontal myoseptum depends in part upon formation of acetylcholine receptor clusters and this was found to be disorganized upon depletion of zfPGRN-A. PGRN reversed the effects of zfPGRN-A knockdown, but a related gene, zfPGRN-1, was without effect. Both knockdown of TDP-43 or FUS, as well as expression of humanTDP-43 and FUS mutants results in MN abnormalities that are reversed by co-expression of hPGRN mRNA. Neither TDP-43 nor FUS reversed MN phenotypes caused by the depletion of PGRN. Thus TDP-43 and FUS lie upstream of PGRN in a gene complementation pathway. The ability of PGRN to override TDP-43 and FUS neurotoxicity due to partial loss of function or mutation in the corresponding genes may have therapeutic relevance.


Comparison of the performance of the Beck Depression Inventory-II and the Center for Epidemiologic Studies-Depression Scale in Arab adolescents.
Few instruments have been established as valid and reliable to screen for depression among Arab adolescents. The purpose of this study was to examine and compare the performance of two of the most widely used depression screening instruments, the Beck Depression Inventory-II (BDI-II) and the Center for Epidemiologic Studies-Depression scale (CES-D), with Arab adolescents. A nationwide school survey was conducted in Jordan. A total of 3,292 adolescents (1,766 females; 54%) aged 13-17 years completed and returned the survey that included the BDI-II, CES-D, sociodemographics, and health information. Comparisons were made between the BDI-II and CES-D on internal consistency and the reported prevalence of depression in the whole population and subgroups. Multivariate ordinary least squares and logistic regressions were used to assess factors associated with adolescent depression. Agreement regarding recommended cutoffs was also examined using Cohen's k. Depression prevalence was significantly higher with the CES-D compared to the BDI-II among the same set of sample subgroups. Depression scores from both instruments showed different statistical associations with established risk factors for adolescent depression. The two instruments showed a moderate agreement (kappa = 0.55), indicating that the instruments do not completely identify the same cases. Different cutoff scores of the CES-D seemed to perform better for different age groups. Our results varied systematically as a function of the measure used to identify depression prevalence. Caution in the interpretation of associations of depression scores with risk factors is required, as associations may be measurement artifacts. However, given the limited availability of mental health care resources in Arab countries, screening instruments like the BDI-II and CES-D may be a critical first step in preliminarily identifying cases, albeit neither can replace the clinical interview. We cautiously recommend using the CES-D with the risk of over diagnosing, but with the benefit of finding issues which are not typically addressed when there is a lack of mental health services. With the growing social and political unrest in Arab countries, increasing depression rates over time is expected to be a major public health issue. Methodical consideration for how to invest in community-based screening is warranted.


Confirmation of direct angiotensin formation by kallikrein.
This study was undertaken to confirm our previous preliminary observation that hog pancreas kallikrein (EC 22.214.171.124) directly liberated an angiotensin-like substance from human plasma protein Cohn fraction IV-4 at an acidic pH of 4.0-5.0. First, the possibility of proangiotensin or des-Asp1-angiotensin being the pressor substance was ruled out by t.l.c. Secondly, the pressor substance was purified by Sephadex G-25 and Bio-Gel P-2 gel filtration, and finally by high-performance liquid chromatography. The amino acid composition of the isolated pressor substance (residues/mol) was: Asp, 1.03; Val, 1.03; Ile, 1.00; Tyr, 0.69; Phe, 1.04; His, 0.91; Arg, 0.86; Pro, 0.86. This composition was identical with that of angiotensin. Since the reaction mixture was not contaminated with common proteolytic enzymes, such as trypsin, chymotrypsin, renin, cathepsin D and proangiotensin-converting enzyme, and other enzymes activated by kallikrein, it is clear that hog kallikrein directly produces angiotensin in vitro.


A data standard for sourcing fit-for-purpose biological samples in an integrated virtual network of biobanks.
Human tissue biobanks are at the epicenter of clinical research, responsible for providing both clinical samples and annotated data. There is a need for large numbers of samples to provide statistical power to research studies, especially since treatment and diagnosis are becoming ever more personalized. A single biobank cannot provide sufficient numbers of samples to capture the full spectrum of any disease. Currently there is no infrastructure in the United Kingdom (UK) to integrate biobanks. Therefore the National Cancer Research Institute (NCRI) Confederation of Cancer Biobanks (CCB) Working Group 3 looked to establish a data standard to enable biobanks to communicate about the samples they hold and so facilitate the formation of an integrated national network of biobanks. The Working Group examined the existing data standards available to biobanks, such as the MIABIS standard, and compared these to the aims of the working group. The CCB-developed data standard has brought many improvements: (1) Where existing data standards have been developed, these have been incorporated, ensuring compatibility with other initiatives; (2) the standard was written with the expectation that it will be extended for specific disease areas, such as the Breast Cancer Campaign Tissue Bank (BCCTB) and the Strategic Tissue Repository Alliances Through Unified Methods (STRATUM) project; and (3) biobanks will be able to communicate about specific samples, as well as aggregated statistics. The development of this data standard will allow all biobanks to integrate and share information about the samples they hold, facilitating the possibility of a national portal for researchers to find suitable samples for research. In addition, the data standard will allow other clinical services, such as disease registries, to communicate with biobanks in a standardized format allowing for greater cross-discipline data sharing.


Analysis of beverages for Hg, As, Pb, and Cd with a field portable X-ray fluorescence analyzer.
Analytical capabilities of a handheld X-ray tube analyzer for analysis of beverages were evaluated. Sets of standard solutions for the elements Hg, As, Pb, and Cd were prepared with mass fractions up to 5000 mg/kg. A thirst quencher beverage was spiked with these elements up to mass fractions of 2500 mg/kg. Portions of these solutions were placed in standard X-ray fluorescence (XRF) cells, as well as the original container, and analyzed by using a field portable Innov-X alpha-6000s XRF tube-type analyzer. Uncorrected analyzer output usually yielded qualitative or semiquantitative results for the spiked beverages in X-ray cells. Average correction factors applied to analyzer output yielded accurate (in terms of z-scores) quantitative results for As above 20 mg/kg and qualitative or semiquantitative results for the other elements. Weighted quadratic fit calibrations provided accurate quantitative or semiquantitative results for all elements at levels above 20 mg/kg. The instrument's preset X-ray overlap correction algorithm worked well for the beverage spiked with all four elements. Spiked beverages analyzed through the wall of the original polyethylene terephthalate container produced accurate results within measurement uncertainties after application of "container wall" correction factors.


Frequency-based heuristics for material perception.
People often make rapid visual judgments of the properties of surfaces they are going to walk on or touch. How do they do this when the interactions of illumination geometry with 3-D material structure and object shape result in images that inverse optics algorithms cannot resolve without externally imposed constraints? A possibly effective strategy would be to use heuristics based on information that can be gleaned rapidly from retinal images. By using perceptual scaling of a large sample of images, combined with correspondence and canonical correlation analyses, we discovered that material properties, such as roughness, thickness, and undulations, are characterized by specific scales of luminance variations. Using movies, we demonstrate that observers' percepts of these 3-D qualities vary continuously as a function of the relative energy in corresponding 2-D frequency bands. In addition, we show that judgments of roughness, thickness, and undulations are predictably altered by adaptation to dynamic noise at the corresponding scales. These results establish that the scale of local 3-D structure is critical in perceiving material properties, and that relative contrast at particular spatial frequencies is important for perceiving the critical 3-D structure from shading cues, so that cortical mechanisms for estimating material properties could be constructed by combining the parallel outputs of sets of frequency-selective neurons. These results also provide methods for remote sensing of material properties in machine vision, and rapid synthesis, editing and transfer of material properties for computer graphics and animation.


Reemerging H5N1 influenza viruses in Hong Kong in 2002 are highly pathogenic to ducks.
Waterfowl are the natural reservoir of all influenza A viruses, which are usually nonpathogenic in wild aquatic birds. However, in late 2002, outbreaks of highly pathogenic H5N1 influenza virus caused deaths among wild migratory birds and resident waterfowl, including ducks, in two Hong Kong parks. In February 2003, an avian H5N1 virus closely related to one of these viruses was isolated from two humans with acute respiratory distress, one of whom died. Antigenic analysis of the new avian isolates showed a reactivity pattern different from that of H5N1 viruses isolated in 1997 and 2001. This finding suggests that significant antigenic variation has recently occurred among H5N1 viruses. We inoculated mallards with antigenically different H5N1 influenza viruses isolated between 1997 and 2003. The new 2002 avian isolates caused systemic infection in the ducks, with high virus titers and pathology in multiple organs, particularly the brain. Ducks developed acute disease, including severe neurological dysfunction and death. Virus was also isolated at high titers from the birds' drinking water and from contact birds, demonstrating efficient transmission. In contrast, H5N1 isolates from 1997 and 2001 were not consistently transmitted efficiently among ducks and did not cause significant disease. Despite a high level of genomic homology, the human isolate showed striking biological differences from its avian homologue in a duck model. This is the first reported case of lethal influenza virus infection in wild aquatic birds since 1961.


Site-specific effects of diselenide bridges on the oxidative folding of a cystine knot peptide, omega-selenoconotoxin GVIA.
Structural and functional studies of small, disulfide-rich peptides depend on their efficient chemical synthesis and folding. A large group of peptides derived from animals and plants contains the Cys pattern C-C-CC-C-C that forms the inhibitory cystine knot (ICK) or knottin motif. Here we report the effect of site-specific incorporation of pairs of selenocysteine residues on oxidative folding and the functional activity of omega-conotoxin GVIA, a well-characterized ICK-motif peptidic antagonist of voltage-gated calcium channels. Three selenoconotoxin GVIA analogues were chemically synthesized; all three folded significantly faster in the glutathione-based buffer compared to wild-type GVIA. One analogue, GVIA[C8U,C19U], exhibited significantly higher folding yields. A recently described NMR-based method was used for mapping the disulfide connectivities in the three selenoconotoxin analogues. The diselenide-directed oxidative folding of selenoconotoxins was predominantly driven by amino acid residue loop sizes formed by the resulting diselenide and disulfide cross-links. Both in vivo and in vitro activities of the analogues were assessed; the block of N-type calcium channels was comparable among the analogues and wild-type GVIA, suggesting that the diselenide replacement did not affect the bioactive conformation. Thus, diselenide substitution may facilitate oxidative folding of pharmacologically diverse ICK peptides. The diselenide replacement has been successfully applied to a growing number of bioactive peptides, including alpha-, mu-, and omega-conotoxins, suggesting that the integrated oxidative folding of selenopeptides described here may prove to be a general approach for efficient synthesis of diverse classes of disulfide-rich peptides.


Generation and characterization of polyclonal antibody against part of immunoglobulin constant heavy υ chain of goose.
Immunoglobulin Y (abbreviated as IgY) is a type of immunoglobulin that is the major antibody in bird, reptile, and lungfish blood. IgY consists of two light (λ) and two heavy (υ) chains. In the present study, polyclonal antibody against IgYFc was generated and evaluated. rIgYCυ3/Cυ4 was expressed in Escherichia coli, purified and utilized to raise polyclonal antibody in rabbit. High affinity antisera were obtained, which successfully detected the antigen at a dilution of 1:204,800 for ELISA assay. The antibody can specifically recognize both rIgYCυ3/Cυ4 and native IgY by Western bolt analysis. Furthermore, the serum of Grus japonensis or immunoglobulin of chicken, duck, turkey, and silkie samples and dynamic changes of serum GoIgY after immunogenicity with GPV-VP3-virus-like particles (GPV-VP3-VLPs) can be detected with the anti-GoIgYFc polyclonal antibody. These results suggested that the antibody is valuable for the investigation of biochemical properties and biological functions of GoIgY.


Intercalating TOP2 Poisons Attenuate Topoisomerase Action at Higher Concentrations.
Topoisomerase II (TOP2) poisons are effective cytotoxic anticancer agents that stabilize the normally transient TOP2-DNA covalent complexes formed during the enzyme reaction cycle. These drugs include etoposide, mitoxantrone, and the anthracyclines doxorubicin and epirubicin. Anthracyclines also exert cell-killing activity via TOP2-independent mechanisms, including DNA adduct formation, redox activity, and lipid peroxidation. Here, we show that anthracyclines and another intercalating TOP2 poison, mitoxantrone, stabilize TOP2-DNA covalent complexes less efficiently than etoposide, and at higher concentrations they suppress the formation of TOP2-DNA covalent complexes, thus behaving as TOP2 poisons at low concentration and inhibitors at high concentration. We used induced pluripotent stem cell (iPSC)-derived human cardiomyocytes as a model to study anthracycline-induced damage in cardiac cells. Using immunofluorescence, our study is the first to demonstrate the presence of topoisomerase IIβ (TOP2B) as the only TOP2 isoform in iPSC-derived cardiomyocytes. In these cells, etoposide robustly induced TOP2B covalent complexes, but we could not detect doxorubicin-induced TOP2-DNA complexes, and doxorubicin suppressed etoposide-induced TOP2-DNA complexes. In vitro, etoposide-stabilized DNA cleavage was attenuated by doxorubicin, epirubicin, or mitoxantrone. Clinical use of anthracyclines is associated with cardiotoxicity. The observations in this study have potentially important clinical consequences regarding the effectiveness of anticancer treatment regimens when TOP2-targeting drugs are used in combination. These observations suggest that inhibition of TOP2B activity, rather than DNA damage resulting from TOP2 poisoning, may play a role in doxorubicin cardiotoxicity. SIGNIFICANCE STATEMENT: We show that anthracyclines and mitoxantrone act as topoisomerase II (TOP2) poisons at low concentration but attenuate TOP2 activity at higher concentration, both in cells and in in vitro cleavage experiments. Inhibition of type II topoisomerases suppresses the action of other drugs that poison TOP2. Thus, combinations containing anthracyclines or mitoxantrone and etoposide may reduce the activity of etoposide as a TOP2 poison and thus reduce the efficacy of drug combinations.


Identification of new genes associated with breast cancer progression by gene expression analysis of predefined sets of neoplastic tissues.
Gene expression profiles were studied by microarray analysis in 2 sets of archival breast cancer tissues from patients with distinct clinical outcome. Seventy-seven differentially expressed genes were identified when comparing 30 cases with relapse and 30 cases without relapse within 72 months from surgery. These genes had a specific ontological distribution and some of them have been linked to breast cancer in previous studies: AIB1, the two keratin genes KRT5 and KRT15, RAF1, WIF1 and MSH6. Seven out of 77 differentially expressed genes were selected and analyzed by qRT-PCR in 127 cases of breast cancer. The expression levels of 6 upregulated genes (CKMT1B, DDX21, PRKDC, PTPN1, SLPI, YWHAE) showed a significant association to both disease-free and overall survival. Multivariate analysis using the significant factors (i.e., estrogen receptor and lymph node status) as covariates confirmed the association with survival. There was no correlation between the expression level of these genes and other clinical parameters. In contrast, SERPINA3, the only downregulated gene examined, was not associated with survival, but correlated with steroid receptor status. An indirect validation of our genes was provided by calculating their association with survival in 3 publicly available microarray datasets. CKMT1B expression was an independent prognostic marker in all 3 datasets, whereas other genes confirmed their association with disease-free survival in at least 1 dataset. This work provides a novel set of genes that could be used as independent prognostic markers and potential drug targets for breast cancer.


Pain inhibition by optogenetic activation of specific anterior cingulate cortical neurons.
Cumulative evidence from both humans and animals suggests that the anterior cingulate cortex (ACC) is important for pain-related perception, and thus a likely target for pain relief therapy. However, use of existing electrode based ACC stimulation has not significantly reduced pain, at least in part due to the lack of specificity and likely co-activation of both excitatory and inhibitory neurons. Herein, we report a dramatic reduction of pain behavior in transgenic mice by optogenetic stimulation of the inhibitory neural circuitry of the ACC expressing channelrhodopsin-2. Electrophysiological measurements confirmed that stimulation of ACC inhibitory neurons is associated with decreased neural activity in the ACC. Further, a distinct optogenetic stimulation intensity and frequency-dependent inhibition of spiking activity in the ACC was observed. Moreover, we confirmed specific electrophysiological responses from different neuronal units in the thalamus, in response to particular types of painful stimuli (i,e., formalin injection, pinch), which we found to be modulated by optogenetic control of the ACC inhibitory neurons. These results underscore the inhibition of the ACC as a clinical alternative in inhibiting chronic pain, and leads to a better understanding of the pain processing circuitry of the cingulate cortex.


Bupropion hydrochloride in attention deficit disorder with hyperactivity.
This is a multisite, double-blind, placebo-controlled trial to determine the safety and efficacy of bupropion in the treatment of children with attention deficit disorder with hyperactivity (ADDH). In a four-center, double-blind comparison of bupropion (n = 72) and placebo (n = 37), children aged 6 to 12 years meeting DSM-III criteria for ADDH were randomized to receive bupropion 3 to 6 mg/kg per day or placebo, administered twice daily, at 7 A.M. and 7 P.M. Measures of efficacy included the Conners Parent and Teacher Questionnaires (93-item, 39-item, and 10 item), Clinical Global Impressions Scales of Severity and Improvement, the Sternberg Short-Term Memory Task, and the Continuous Performance Test. Screen and posttreatment physical examinations, electrocardiograms, electroencephalograms, and clinical laboratory evaluations were performed. Height, weight, and vital signs were measured and adverse experiences were assessed weekly. A significant treatment effect, apparent as early as day 3, was present for both conduct problems and hyperactivity on the Conners 10-item and 39-item teacher's checklist, and at day 28 for conduct problems and restless-impulsive behavior on the 93-item parent questionnaire. Findings were of smaller magnitude for parent ratings than teacher ratings. Significant treatment effects were present on both the Continuous Performance Test and memory retrieval test. Effect sizes of bupropion/placebo differences for teacher and parent ratings in this study were somewhat smaller than for standard stimulant drugs used to treat ADDH. Bupropion appeared to be well tolerated in most children. Dermatological reactions were twice as frequent in the drug group as the placebo group, with four reactions involving rash and urticaria that were serious enough to require discontinuation of medication. Bupropion may be a useful addition to available treatments for ADDH. Comparative trials with such standard drugs as methylphenidate are warranted to determine the relative clinical merits of bupropion.


Stimulated Axion Decay in Superradiant Clouds around Primordial Black Holes.
The superradiant instability can lead to the generation of extremely dense axion clouds around rotating black holes. We show that, despite the long lifetime of the QCD axion with respect to spontaneous decay into photon pairs, stimulated decay becomes significant above a minimum axion density and leads to extremely bright lasers. The lasing threshold can be attained for axion masses μ≳10^{-8} eV, which implies superradiant instabilities around spinning primordial black holes with mass ≲0.01 M_{⊙}. Although the latter are expected to be nonrotating at formation, a population of spinning black holes may result from subsequent mergers. We further show that lasing can be quenched by Schwinger pair production, which produces a critical electron-positron plasma within the axion cloud. Lasing can nevertheless restart once annihilation lowers the plasma density sufficiently, resulting in multiple laser bursts that repeat until the black hole spins down sufficiently to quench the superradiant instability. In particular, axions with a mass ∼10^{-5} eV and primordial black holes with mass ∼10^{24} kg, which may account for all the dark matter in the Universe, lead to millisecond bursts in the GHz radio-frequency range, with peak luminosities ∼10^{42} erg/s, suggesting a possible link to the observed fast radio bursts.


Standards for thoracic surgical oncology in a single-payer healthcare system.
Through systematic literature review and a consensus-based approach from an expert panel, standards on the organization for delivering thoracic cancer surgery in a single-payer healthcare environment were developed. Thirty-two studies and six organizational reports were identified. Results from 32 studies showed a trend toward higher volumes and improved patient outcomes, and six consensus reports provided recommendations on thoracic care standards. Thoracic surgical oncology standards in a single-payer healthcare system were developed. The benefits associated with the implementation of thoracic cancer surgery standards should result in increased regionalization of care, improved processes of care, and better patient outcomes.