Genotyping-by-Sequencing and Its Application to Oat Genomic Research.
Genotyping-by-sequencing (GBS) has emerged as a useful genomic approach for sampling genome-wide genetic variation, performing genome-wide association mapping, and conducting genomic selection. It is a combined one-step process of SNP marker discovery and genotyping through genome reduction with restriction enzymes and SNP calling with or without a sequenced genome. This approach has the advantage of being rapid, high throughput, cost effective, and applicable to organisms without sequenced genomes. It has been increasingly applied to generate SNP genotype data for plant genetic and genomic studies. To facilitate a wider GBS application, particularly in oat genetic and genomic research, we describe the GBS approach, review the current applications of GBS in plant species, and highlight some applications of GBS to oat research. We also discuss issues in various applications of GBS and provide some perspectives in GBS research. Recent developments of bioinformatics pipelines in high-quality SNP discovery for polyploid crops will enhance the application of GBS to oat genetic and genomic research.


Gregarious cyanobacteria.
Huber and collaborators reported in this issue of Environmental Microbiology about freshwater picocyanobacteria that showed phenotypic plasticity in the sense that they appeared as single cells as well as in aggregates. The authors suggested that aggregation might be an inducible defense as a response to the presence of grazers. This has been described for eukaryotic phytoplankton and for the cyanobacterium Microcystis but thus far not for picocyanobacteria. Although inducible defense as an explanation is an attractive possibility, it is also problematic. Aggregation is common among cyanobacteria and it offers many advantages as compared with a free-living lifestyle. Here these advantages are highlighted and the possibility of inducible defense is critically assessed.


A rule-based model of barley morphogenesis, with special respect to shading and gibberellic acid signal transduction.
Functional-structural plant models (FSPM) constitute a paradigm in plant modelling that combines 3D structural and graphical modelling with the simulation of plant processes. While structural aspects of plant development could so far be represented using rule-based formalisms such as Lindenmayer systems, process models were traditionally written using a procedural code. The faithful representation of structures interacting with functions across scales, however, requires a new modelling formalism. Therefore relational growth grammars (RGG) were developed on the basis of Lindenmayer systems. In order to implement and test RGG, a new modelling language, the eXtended L-system language (XL) was created. Models using XL are interpreted by the interactive, Java-based modelling platform GroIMP. Three models, a semi-quantitative gibberellic acid (GA) signal transduction model, and a phytochrome-based shade detection and object avoidance model, both coupled to an existing morphogenetic structural model of barley (Hordeum vulgare L.), serve as examples to demonstrate the versatility and suitability of RGG and XL to represent the interaction of diverse biological processes across hierarchical scales. The dynamics of the concentrations in the signal transduction network could be modelled qualitatively and the phenotypes of GA-response mutants faithfully reproduced. The light model used here was simple to use yet effective enough to carry out local measurement of red:far-red ratios. Suppression of tillering at low red:far-red ratios could be simulated. The RGG formalism is suitable for implementation of multi-scaled FSPM of plants interacting with their environment via hormonal control. However, their ensuing complexity requires careful design. On the positive side, such an FSPM displays knowledge gaps better thereby guiding future experimental design.


Characterization of myelin proteolipid mRNAs in normal and jimpy mice.
A clone specific for the rat myelin proteolipid protein (PLP) was isolated from a cDNA library made in pUC18 from 17-day-old rat brain stem mRNA. This clone corresponded to the carboxyl-terminal third of the PLP-coding region. The clone was used to identify PLP-specific mRNAs in mouse brain and to establish the time course of PLP mRNA expression during mouse brain development. Three PLP-specific mRNAs were seen, approximately 1,500, 2,400, and 3,200 bases in length, of which the largest was the most abundant. During brain development, the maximal period of PLP mRNA expression was from 14 to 25 days of age, and this was a similar time course to that for myelin basic protein mRNA expression. When the jimpy mouse, an X-linked dysmyelination mutant, was studied for PLP mRNA expression, low levels of PLP mRNA were seen which were approximately 5% of wild-type levels at 20 days of age. When jimpy brain RNA was analyzed by Northern blotting, the PLP-specific mRNA was shown to be 100 to 200 bases shorter than the wild-type PLP-specific mRNA. This size difference was seen in the two major PLP mRNAs, and it did not result from a loss of polyadenylation of these mRNAs.


A new mutation in the connexin 32 gene was found in Charcot- Marie-Tooth disease in Chinese patients.
To investigate the characteristics of gene mutations of connexin 32 exon 2 in Charcot-Marie-Tooth disease in Chinese patients. Screening for connexin 32 gene mutation was conducted in 6 unrelated CMT1 patients without duplication and 10 unrelated CMT2 patients. Mobility shift of exon 2 was analyzed by SSCP and further confirmed by sequencing. The PCR products were cut by appropriate restricted enzyme in 50 normal controls. One missense mutation at nucleotides 62(G-->A) was found in a CMT1 patient. 50 normal controls were analyzed by the enzyme HaeIII and no abnormality was found. This proved that the mutation was the cause of disease. This mutation has not been reported previously. A proportion of CMTX patients may exist in the group of CMT1 patients in China.


Effective source size, yield and beam profile from multi-layered bremsstrahlung targets.
Modern conformal radiotherapy benefits from heterogeneous dose delivery using scanned narrow bremsstrahlung beams of high energy in combination with dynamic double focused multi-leaf collimation and purging magnets. When using a purging magnet to remove electrons and positrons the target space is limited and unorthodox thin multi-layered targets are needed. A computational technique has therefore been developed to determine the forward yield and the angular distributions of the bremsstrahlung beam as well as the size and location of the effective and the virtual photon point source for arbitrary multi-layer bremsstrahlung targets. The Gaussian approximation of the diffusion equation for the electrons has been used and convolved with the bremsstrahlung production process. For electrons with arbitrary emittance impinging on targets of any multi-layer and atomic number combination, the model is well applicable, at least for energies in the range 1-100 MeV. The intrinsic bremsstrahlung photon profile has been determined accurately by deconvolving the electron multiple scattering process from thin experimental beryllium target profiles. For electron pencil beams incident on a target of high density and atomic number such as tungsten, the size of the effective photon source stays at around a tenth of a millimetre. The effective photon source for low-Z materials such as Be, C and Al is located at depths from 3-7 mm in the target, decreasing with increasing atomic number. The effective photon source at off-axis positions then moves out considerably from the central axis, which should be considered when aligning collimators. For high-Z materials such as tungsten, the location of the effective photon source is at a few tenths of a millimetre deep. The virtual photon point source is located only a few tenths of a millimetre upstream of the effective photon source both for high- and low-Z materials. For 50 MeV electrons incident on multi-layered full range targets the radial energy fluence distributions will have a full width at half maximum (FWHM) of 80 to 100 mm at 1 m from the target. The best target composition made of two layers when the space is limited to 15 mm was found to be 9 mm-Be followed by 6 mm W. A thin beryllium target (approximately 3 mm) results in a high-intensity bremsstrahlung lobe with a FWHM of about 35 mm at the isocentre. Interestingly, the forward dose rate in such a beam is as high as 62% of the maximum achievable with an optimal target design, even if on average only 1 MeV is lost by the electrons.


Assessment and treatment of the failing right heart: considerations for transplantation referral.
Long considered an insignificant chamber, the right ventricle has been recently under spotlight, especially with emerging data supporting its critical role in disease progression. The right ventricle is a triangular heart chamber with complex physiology and pathophysiology. In this review, we discuss the normal physiology of the right ventricle, right ventricular failure, and recent advances in treatment with emphasis on optimal timing for transplantation referral.


Combining acoustic and electrical speech processing: Iowa/Nucleus hybrid implant.
In this paper we test the concept of combining electrical stimulation for high-frequency sound with acoustic hearing for low-frequency information in the same ear. In addition, we test whether residual hearing can be preserved when an electrode is placed up to 10 mm into the inner ear, and whether the site of electrical stimulation influences speech perception. Nine post-lingual adults with severe high-frequency hearing impairment were recruited to participate in the study. A single-subject clinical trial design was employed. A unique six-channel cochlear implant was designed for this clinical trial. The intracochlear electrodes were either 6 or 10 mm in length based on a Nucleus CI-24 multichannel implant. Monosyllabic word understanding and consonant identification in a recorded sound-only condition were used to assess changes in speech perception. Follow-up was > 12 months. Acoustic hearing was preserved in all nine subjects. Preoperative monosyllabic word and sentence scores were unchanged in all subjects following implantation. A 30-40% improvement in consonant recognition occurred with the 10-mm electrode. The 10-mm electrode subjects were able to understand 83-90% of the monosyllabic words using the implant plus binaural hearing aids. Scores were more than doubled when compared to preoperative scores achieved with hearing aids only. The human ear has the capability to integrate both acoustic and high-frequency electrically processed speech information. Placement of a short 10-mm electrode does not appear to damage residual low-frequency inner ear hair cell function, interfere with the micro-mechanics of normal cochlear vibration or decrease residual speech perception. The improvement in speech recognition was due primarily to the increased perception of higher-frequency consonantal speech cues. Such a device can provide a substantial benefit in terms of speech understanding to those with severe high-frequency hearing loss, while still maintaining the benefits of the residual lower-frequency acoustic hearing. The position of the electrode and the site of frequency information within the cochlea are shown to be important factors in the success of such a device.


Fibroblast Growth Factor Receptor 2 Signaling in Breast Cancer.
Fibroblast growth factor receptor 2 (FGFR2) is a membrane-spanning tyrosine kinase that mediates signaling for FGFs. Recent studies detected various point mutations of FGFR2 in multiple types of cancers, including breast cancer, lung cancer, gastric cancer, uterine cancer and ovarian cancer, yet the casual relationship between these mutations and tumorigenesis is unclear. Here we will discuss possible interactions between FGFR2 signaling and several major pathways through which the aberrantly activated FGFR2 signaling may result in breast cancer development. We will also discuss some recent developments in the discovery and application of therapies and strategies for breast cancers by inhibiting FGFR2 activities.


Experimentation or observation? Of the self alone or the natural world?
One important lesson of Roberts' target article may be potentially obscured for some by the title's reference to "self-experimentation." At the core of this work, the key investigative resource is sustained and systematic observation, not experimentation, and it is deployed in a fashion not necessarily restricted to self-examination. There is an important reminder here of a strategically important, but neglected, relationship between observation and experiment.


Bacteriologic survey of black rhinoceros (Diceros bicornis).
A bacteriological survey was carried out on 30 black rhinoceros (Diceros bicornis) of which 23 were newly captured and seven were captive. A beta haemolytic Streptococcus, group L was found in skin lesions and various wounds, causing septicaemia and death in two animals. Staphylococcus aureus was found in 3 rhinoceros, and caused the death of one. The bacteria isolated often proved resistant to penicillin. Streptomycin is recommended for treatment. Sixteen other bacteria sp. were isolated, and apart from a Salmonella sp. none were considered to be specific pathogens.


Cloning of estrogen-regulated messenger RNA sequences from human breast cancer cells.
A complementary DNA library was constructed from RNA of estrogen-stimulated MCF-7 cells and screened for estrogen-regulated sequences. Four different messenger RNA sequences of varying abundance were isolated. Two of the sequences (pNR-3 and pNR-4) were induced approximately 2-fold, while the other two (pNR-1 and pNR-2) were induced at least 8-fold. The induction of both pNR-1 and pNR-2 requires similar physiological concentrations of estradiol and is near maximal at 10(-10) M. An increase in the levels of the RNAs is seen after 30 min of estrogen treatment, but pNR-1 reaches its maximal concentration faster than pNR-2. pNR-1 and pNR-2 were not expressed in all human breast cancer cell lines tested. pNR-1 was expressed and regulated by estrogen in the estrogen receptor-positive cell lines, MCF-7, T-47D, and ZR 75, whereas pNR-2 was not expressed in the T-47D cell line. pNR-1 and pNR-2 were not detected in two estrogen receptor-negative cell lines (BT20 and HBL 100). As the proliferation of the MCF-7, T47D, and ZR 75 cell lines is stimulated by estradiol, pNR-1 may provide a useful marker of hormone-responsive breast cancer.


Direct monitoring of the strand passage reaction of DNA topoisomerase II triggers checkpoint activation.
By necessity, the ancient activity of type II topoisomerases co-evolved with the double-helical structure of DNA, at least in organisms with circular genomes. In humans, the strand passage reaction of DNA topoisomerase II (Topo II) is the target of several major classes of cancer drugs which both poison Topo II and activate cell cycle checkpoint controls. It is important to know the cellular effects of molecules that target Topo II, but the mechanisms of checkpoint activation that respond to Topo II dysfunction are not well understood. Here, we provide evidence that a checkpoint mechanism monitors the strand passage reaction of Topo II. In contrast, cells do not become checkpoint arrested in the presence of the aberrant DNA topologies, such as hyper-catenation, that arise in the absence of Topo II activity. An overall reduction in Topo II activity (i.e. slow strand passage cycles) does not activate the checkpoint, but specific defects in the T-segment transit step of the strand passage reaction do induce a cell cycle delay. Furthermore, the cell cycle delay depends on the divergent and catalytically inert C-terminal region of Topo II, indicating that transmission of a checkpoint signal may occur via the C-terminus. Other, well characterized, mitotic checkpoints detect DNA lesions or monitor unattached kinetochores; these defects arise via failures in a variety of cell processes. In contrast, we have described the first example of a distinct category of checkpoint mechanism that monitors the catalytic cycle of a single specific enzyme in order to determine when chromosome segregation can proceed faithfully.


Mind-Body Practices and the Self: Yoga and Meditation Do Not Quiet the Ego but Instead Boost Self-Enhancement.
Mind-body practices enjoy immense public and scientific interest. Yoga and meditation are highly popular. Purportedly, they foster well-being by curtailing self-enhancement bias. However, this "ego-quieting" effect contradicts an apparent psychological universal, the self-centrality principle. According to this principle, practicing any skill renders that skill self-central, and self-centrality breeds self-enhancement bias. We examined those opposing predictions in the first tests of mind-body practices' self-enhancement effects. In Experiment 1, we followed 93 yoga students over 15 weeks, assessing self-centrality and self-enhancement bias after yoga practice (yoga condition, n = 246) and without practice (control condition, n = 231). In Experiment 2, we followed 162 meditators over 4 weeks (meditation condition: n = 246; control condition: n = 245). Self-enhancement bias was higher in the yoga (Experiment 1) and meditation (Experiment 2) conditions, and those effects were mediated by greater self-centrality. Additionally, greater self-enhancement bias mediated mind-body practices' well-being benefits. Evidently, neither yoga nor meditation fully quiet the ego; to the contrary, they boost self-enhancement.


The conserved metalloprotease invadolysin is present in invertebrate haemolymph and vertebrate blood.
We identified invadolysin, a novel essential metalloprotease, for functions in chromosome structure, cell proliferation and migration. Invadolysin also plays an important metabolic role in insulin signalling and is the only protease known to localise to lipid droplets, the main lipid storage organelle in the cell. In silico examination of the protein sequence of invadolysin predicts not only protease and lipase catalytic motifs, but also post-translational modifications and the secretion of invadolysin. Here we show that the protease motif of invadolysin is important for its role in lipid accumulation, but not in glycogen accumulation. The lipase motif does not appear to be functionally important for the accumulation of lipids or glycogen. Post-translational modifications likely contribute to modulating the level, localisation or activity of invadolysin. We identified a secreted form of invadolysin in the soluble fraction of invertebrate hemolymph (where we observe sexually dimorphic forms) and also vertebrate plasma, including in the extracellular vesicle fraction. Biochemical analysis for various post-translational modifications demonstrated that secreted invadolysin is both N- and O-glycosylated, but not apparently GPI-linked. The discovery of invadolysin in the extracellular milieu suggests a role for invadolysin in normal organismal physiology.


The co-chaperone and reductase ERdj5 facilitates rod opsin biogenesis and quality control.
Mutations in rhodopsin, the light-sensitive protein of rod cells, are the most common cause of autosomal dominant retinitis pigmentosa (ADRP). Many rod opsin mutations, such as P23H, lead to misfolding of rod opsin with detrimental effects on photoreceptor function and viability. Misfolded P23H rod opsin and other mutations in the intradiscal domain are characterized by the formation of an incorrect disulphide bond between C185 and C187, as opposed to the correct and highly conserved C110-C187 disulphide bond. Therefore, we tested the hypothesis that incorrect disulphide bond formation might be a factor that affects the biogenesis of rod opsin by studying wild-type (WT) or P23H rod opsin in combination with amino acid substitutions that prevent the formation of incorrect disulphide bonds involving C185. These mutants had altered traffic dynamics, suggesting a requirement for regulation of disulphide bond formation/reduction during rod opsin biogenesis. Here, we show that the BiP co-chaperone and reductase protein ERdj5 (DNAJC10) regulates this process. ERdj5 overexpression promoted the degradation, improved the endoplasmic reticulum mobility and prevented the aggregation of P23H rod opsin. ERdj5 reduction by shRNA delayed rod opsin degradation and promoted aggregation. The reductase and co-chaperone activity of ERdj5 were both required for these effects on P23H rod opsin. Furthermore, mutations in these functional domains acted as dominant negatives that affected WT rod opsin biogenesis. Collectively, these data identify ERdj5 as a member of the proteostasis network that regulates rod opsin biogenesis and supports a role for disulphide bond formation/reduction in rod opsin biogenesis and disease.


Experience with Esophagogastrointestinal Transit Scintigraphy in the Initial 229 Patients: Multiple Regions of Dysmotility Are Common.
The purpose of this investigation was to review our experience with our comprehensive esophagogastrointestinal transit study in the first 229 patients. This scintigraphic study analyzes the motility of the entire gut, from the esophagus through the rectosigmoid colon. Methods: Data were reviewed for our first 2 y of experience with this examination (184 women and 45 men aged 20-79 y [mean ± SD, 44 ± 16 y]). Patients were referred with symptoms suggestive of a motility disorder. They first swallowed 111In-diethylenetriaminepentaacetic acid in water for the esophageal-swallow study and then 300 mL for a 30-min 111In water-only study, followed by 120 mL of 111In water simultaneously with the solid standardized 99mTc egg-substitute meal. Images and quantification were obtained for esophageal transit, water-only gastric emptying, water-with-solid gastric emptying, small-bowel transit, and colonic transit. Results: Of the 229 patient studies, 45 (20%) were normal. The remaining 184 (80%) had at least 1 region of dysmotility, for a total of 336 regions of abnormal motility. A single region of dysmotility was seen in 92 patients (50%), 2 regions in 50 (27%), 3 regions in 26 (14%), 4 regions in 12 (7%), and 5 regions in 4 (2%). There was a poor correlation between the results of the water-only study and water with the solid meal. Three different patterns of delayed colonic transit were seen. Patient symptoms were often not predictive of the scintigraphic findings. Conclusion: This study highlights the frequent occurrence of dysmotility in more than 1 region of the gastrointestinal tract in patients with a suspected motility disorder and the frequent concurrence of both upper- and lower-tract dysmotility in the same patients. It provides information to referring physicians regarding which motility disorders may be causing the patient symptoms, why the patient is or is not responding to the present therapy, and if and what additional workup and therapy may be needed.


Convection currents in a water calorimeter.
A flexible, temperature-regulated water calorimeter has been constructed containing two pairs of thermistor sensors at depths of 6.23 and 10.0 cm. It may be irradiated by vertical or horizontal beams, and operated at temperatures in the range from 3 to 40 degrees C. When irradiated at 30 degrees C with a vertically downward 19 MeV electron beam, the responses of the proximal and midline thermistors were in accordance with the depth-dose curve. When irradiated horizontally, the initial patterns of temperature rise were the same, but after about 30 s (4 Gy) the rate of temperature rise decreased at the proximal thermistors and increased at the midline thermistors. Shortly after irradiation, the temperature curve and increased at the midline thermistors. Shortly after irradiation, the temperature curve of the midline thermistors crossed that for the proximal thermistors, a pattern that suggested the presence of convection currents. To test this hypothesis, the calorimeter was operated at 4 degrees C. The temperature patterns for horizontal irradiation became the same as those obtained with vertical beams, thus demonstrating the production of convection currents in water at a temperature of 30 degrees C for temperature gradients as small as 10(-3) degrees C cm-1.


Differential Response of Two Human Breast Cancer Cell Lines to the Phenolic Extract from Flaxseed Oil.
Many studies have evidenced that the phenolic components from flaxseed (FS) oil have potential health benefits. The effect of the phenolic extract from FS oil has been evaluated on two human breast cancer cell lines, MCF7 and MDA-MB231, and on the human non-cancerous breast cell line, MCF10A, by SRB assay, cellular death, cell cycle, cell signaling, lipid peroxidation and expression of some key genes. We have evidenced that the extract shows anti-proliferative activity on MCF7 cells by inducing cellular apoptosis, increase of the percentage of cells in G0/G1 phase and of lipid peroxidation, activation of the H2AX signaling pathway, and upregulation of a six gene signature. On the other hand, on the MDA-MB2131 cells we verified only an anti-proliferative activity, a weak lipid peroxidation, the activation of the PI3K signaling pathway and an up-regulation of four genes. Overall these data suggest that the extract has both cytotoxic and pro-oxidant effects only on MCF7 cells, and can act as a metabolic probe, inducing differences in the gene expression. For this purpose, we have performed an interactomic analysis, highlighting the existing associations. From this approach, we show that the phenotypic difference between the two cell lines can be explained through their differential response to the phenolic extract.


Atherothrombosis and plaque heterology: different location or a unique disease?
Formation of unstable plaques frequently results in atherothrombosis, the major cause for ischaemic stroke, myocardial infarction and peripheral arterial disease. Patients who have symptomatic thrombosis in one vascular bed are at increased risk of disease in other beds. However, the development of the disease in carotid, coronary and peripheral arteries may have different pathophysiology suggesting that more complex treatment protocols may have to be designed to reduce plaque development at different locations. In this review we describe the known risk factors, compare the developmental features of coronary and carotid plaque development and determine their association with end-point ischaemic events. Differences are also seen in the genetic contribution to plaque development as well as in the deregulation of gene and protein expression and cellular signal transduction activity of active cells in regions susceptible to thrombosis. Differences between carotid and coronary artery plaque development might help to explain the differences in anatomopathological appearance and risk of rupture.


Predictors of completeness of patients' self-reported personal medication lists and discrepancies with clinic medication lists.
Transfer of medication information during transitions in care is crucial to preventing medication errors. Few studies evaluate patients' self-reported personal medication lists. To assess completeness of personal medication lists and identify factors associated with incomplete personal lists and discrepancies between personal and clinic medication lists. We analyzed patients' personal medication lists at an academic hospital preoperative clinic from January 2010 to October 2010. Completeness of personal medication lists was measured as reporting the name, dose, and frequency for all prescription and nonprescription medications or dietary supplements. Discrepancies between personal and clinic medication lists were measured as omitted medications or differing directions. Among 94 patients meeting inclusion criteria, 82 (87%) personal medication lists were evaluated. Most personal lists were incomplete (56%; 46/82), missing information for at least one medication reported; 94% (77/82) of personal lists had at least one discrepancy with clinic medication lists (median 4 discrepancies per patient list). On multivariate analyses, taking 10 or more medications (adjusted odds ratio [OR] = 3.52; 95% CI = 1.37 to 9.08) and being divorced, widowed, or single (adjusted OR = 3.10; 95% CI = 1.05 to 9.12) were independent predictors of incomplete personal medication lists. Taking 10 or more medications (adjusted OR = 3.44; 95% CI = 1.35 to 8.78) was also associated with higher rates of medication discrepancies. Patients' self-reported personal medication lists are often incomplete and have discrepancies with clinic medication lists. Interventions are needed to improve medication information transfer between patients, providers and healthcare systems.


TET1-mediated epigenetic regulation of tumor necrosis factor-α in trigeminal ganglia contributes to chronic temporomandibular joint pain.
Chronic temporomandibular joint (TMJ) pain profoundly affects patients' quality of life. Trigeminal tumor necrosis factor-α (TNFα) plays a pivotal role in mediating TMJ pain in mice, yet the underlying epigenetic mechanisms remain enigmatic. To unravel these epigenetic intricacies, we employed a multifaceted approach. Hydroxymethylated DNA immunoprecipitation (hMeDIP) and chromatin immunoprecipitation (ChIP) followed by qPCR were employed to investigate the demethylation of TNFα gene (Tnfa) and its regulation by ten-eleven translocation methylcytosine dioxygenase 1 (TET1) in a chronic TMJ pain mouse model. The global levels of 5-hydroxymethylcytosine (5hmc) and percentage of 5hmc at the Tnfa promoter region were measured in the trigeminal ganglia (TG) and spinal trigeminal nucleus caudalis (Sp5C) following complete Freund's adjuvant (CFA) or saline treatment. TET1 knockdown and pain behavioral testing were conducted to ascertain the role of TET1-mediated epigenetic regulation of TNFα in the pathogenesis of chronic TMJ pain. Our finding revealed an increase in 5hmc at the Tnfa promoter region in both TG and Sp5C of CFA-treated mice. TET1 was upregulated in the mouse TG, and the ChIP result showed TET1 direct binding to the Tnfa promoter, with higher efficiency in the CFA-treated group. Immunofluorescence revealed the predominant expression of TET1 in trigeminal neurons. TET1 knockdown in the TG significantly reversed CFA-induced TNFα upregulation and alleviated chronic TMJ pain. In conclusion, our study implicates TET1 as a vital epigenetic regulator contributing to chronic inflammatory TMJ pain via trigeminal TNFα signaling. Targeting TET1 holds promise for epigenetic interventions in TMJ pain management.


Percutaneous innominate vein catheterisation: a new technique for venous access in the critically ill.
Venous access in the critically ill can be a major problem despite the wide variety of methods now available. A new technique is described involving percutaneous catheterisation of the innominate vein via a right second intercostal space approach. It has been successful in five patients over a four-year period. While there have been no major complications, it is advocated for use only when access is necessary for life-saving administration of fluids or drugs. Venous access can be obtained in the majority of patients by a variety of safe and accepted measures such as peripheral venepuncture, central vein catheterisation by the subclavian or internal jugular vein, or by a cutdown. There remains, however, a small subgroup of patients with venous thrombosis from a previous catheter insertion or intravenous drug abuse for whom a number of ingenious methods have been devised. These include cutdowns on the intercostal vein; the middle thyroid vein in infants; and on the inferior epigastric vein, the azygos vein, the iliac vein, and even the inferior vena cava in adults. Access for patients suffering from haemorrhagic shock can be facilitated by multiple catheters in one or more subclavian veins. Even more rarely, these methods may be unsuccessful or contraindicated because of vein thrombosis, infection overlying the insertion site, or patient instability. A new method has been developed allowing percutaneous catheterisation of the innominate vein.


Cell calcium and arterial blood pressure.
In hypertensive humans and animals, intracellular calcium (Ca2+) has been found to be elevated in several cell types, including platelets, red blood cells, and smooth muscle cells. The messenger role of calcium requires its maintenance within cells at very low (submicromolar) ionic concentrations. This transcellular gradient is maintained by the reversible complexing of calcium to specific proteins. Intracellular free calcium can increase through an enhanced influx, release from intracellular stores, or a reduced efflux of calcium from the cell. Calcium channels are found to be increased in activity in some studies of hypertensive animals, but not in others. Sequestration of calcium within the cell has also been implicated in the increased cytosolic calcium in hypertension. Plasma membrane adenosine triphosphatases (ATPases) play a significant role in calcium extrusion; and the activity of these pumps has been found to be decreased in several human and animal studies. A single cause of the cellular calcium alterations in hypertension is not completely clear.


[Photopheresis--anew method for treatment of myasthenia].
Photopheresis with blood lymphocytes UV radiation exposure was for the first time used in the treatment of patients with myasthenia. Twenty-three patients with generalized form of the disease, aged 23-66 years, were treated. Twenty patients (87%) exhibited clinical improvement after a treatment course (4 sessions). One photopheresis session was shown to be more or less beneficial in 16 out of 23 patients (70%). The treatment resulted in strengthening of all the muscles studied, especially of mimic, platysma, biceps, deltoid, illiopsoas and gastrocnemius. Photopheresis was effective not only in a mild and moderate but in severe form of myasthenia as well. Compared to plasmapheresis, photopheresis exerts a more prolonged effect. Clinical improvement correlated with positive changes in immune status of the patients.


Expanding the CRISPR Toolbox in Culicine Mosquitoes: In Vitro Validation of Pol III Promoters.
CRISPR-Cas9-based "gene drive" technologies have been proposed as a novel and effective means of controlling human diseases vectored by mosquitoes. However, more complex designs than those demonstrated to date-and an expanded molecular toolbox with which to build them-will be required to overcome the issues of resistance formation/evolution and drive spatial/temporal limitation. Foreseeing this need, we assessed the sgRNA transcriptional activities of 33 phylogenetically diverse insect Polymerase III promoters using three disease-relevant Culicine mosquito cell lines (Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus). We show that U6 promoters work across species with a range of transcriptional activity levels and find 7SK promoters to be especially promising because of their broad phylogenetic activity. We further show that U6 promoters can be substantially truncated without affecting transcriptional levels. These results will be of great utility to researchers involved in developing the next generation of gene drives.


Are subthreshold alcohol dependence symptoms a risk factor for developing DSM-IV alcohol use disorders? A three-year prospective study of 'diagnostic orphans' in a national sample.
Research suggests that diagnostic orphans (i.e., individuals experiencing only 1-2 criteria for DSM-IV alcohol dependence) may be at increased risk for developing more severe alcohol problems. This study aimed to: (i) investigate the course of diagnostic orphans in the National Epidemiologic Survey on Alcohol and Related Conditions (NESARC), and (ii) explore whether a specific symptom endorsement pattern(s) could identify diagnostic orphans at Wave 1 who remitted or progressed to alcohol dependence at Wave 2. Current drinkers (n = 15,751) were divided into diagnostic groups at Waves 1 and 2: no-alcohol use disorder (AUD); one-criterion orphans, two-criterion orphans, alcohol abuse, or alcohol dependence. Multinomial logistic regression analysis explored the association between diagnostic status at Wave 1 and Wave 2. Chi-square tests investigated differences in the criteria endorsement patterns of diagnostic orphans. Compared to the no-AUD group, one-criterion orphans at Wave 1 were twice as likely to be in the abuse group and four times more likely to be dependent at Wave 2. Two-criterion orphans were three times more likely to be in the abuse group and eight times more likely to have progressed to dependence. Criterion endorsement patterns of diagnostic orphans at baseline did not significantly differentiate between those who remitted and those who progressed to dependence at follow-up. Like previous research, diagnostic orphans are at increased for developing to more severe alcohol problems. Relying solely on the DSM-IV AUD diagnostic criteria, however, may not be sufficient to identify those diagnostic orphans who are at risk for progressing to dependence.


Dietary Supplements and Nutraceuticals under Investigation for COVID-19 Prevention and Treatment.
Coronavirus disease 2019 (COVID-19) has caused global disruption and a significant loss of life. Existing treatments that can be repurposed as prophylactic and therapeutic agents may reduce the pandemic's devastation. Emerging evidence of potential applications in other therapeutic contexts has led to the investigation of dietary supplements and nutraceuticals for COVID-19. Such products include vitamin C, vitamin D, omega 3 polyunsaturated fatty acids, probiotics, and zinc, all of which are currently under clinical investigation. In this review, we critically appraise the evidence surrounding dietary supplements and nutraceuticals for the prophylaxis and treatment of COVID-19. Overall, further study is required before evidence-based recommendations can be formulated, but nutritional status plays a significant role in patient outcomes, and these products may help alleviate deficiencies. For example, evidence indicates that vitamin D deficiency may be associated with a greater incidence of infection and severity of COVID-19, suggesting that vitamin D supplementation may hold prophylactic or therapeutic value. A growing number of scientific organizations are now considering recommending vitamin D supplementation to those at high risk of COVID-19. Because research in vitamin D and other nutraceuticals and supplements is preliminary, here we evaluate the extent to which these nutraceutical and dietary supplements hold potential in the COVID-19 crisis.IMPORTANCE Sales of dietary supplements and nutraceuticals have increased during the pandemic due to their perceived "immune-boosting" effects. However, little is known about the efficacy of these dietary supplements and nutraceuticals against the novel coronavirus (severe acute respiratory syndrome coronavirus 2 [SARS-CoV-2]) or the disease that it causes, CoV disease 2019 (COVID-19). This review provides a critical overview of the potential prophylactic and therapeutic value of various dietary supplements and nutraceuticals from the evidence available to date. These include vitamin C, vitamin D, and zinc, which are often perceived by the public as treating respiratory infections or supporting immune health. Consumers need to be aware of misinformation and false promises surrounding some supplements, which may be subject to limited regulation by authorities. However, considerably more research is required to determine whether dietary supplements and nutraceuticals exhibit prophylactic and therapeutic value against SARS-CoV-2 infection and COVID-19. This review provides perspective on which nutraceuticals and supplements are involved in biological processes that are relevant to recovery from or prevention of COVID-19.


AI-based diagnosis of nuclear cataract from slit-lamp videos.
In ophthalmology, the availability of many fundus photographs and optical coherence tomography images has spurred consideration of using artificial intelligence (AI) for diagnosing retinal and optic nerve disorders. However, AI application for diagnosing anterior segment eye conditions remains unfeasible due to limited standardized images and analysis models. We addressed this limitation by augmenting the quantity of standardized optical images using a video-recordable slit-lamp device. We then investigated whether our proposed machine learning (ML) AI algorithm could accurately diagnose cataracts from videos recorded with this device. We collected 206,574 cataract frames from 1812 cataract eye videos. Ophthalmologists graded the nuclear cataracts (NUCs) using the cataract grading scale of the World Health Organization. These gradings were used to train and validate an ML algorithm. A validation dataset was used to compare the NUC diagnosis and grading of AI and ophthalmologists. The results of individual cataract gradings were: NUC 0: area under the curve (AUC) = 0.967; NUC 1: AUC = 0.928; NUC 2: AUC = 0.923; and NUC 3: AUC = 0.949. Our ML-based cataract diagnostic model achieved performance comparable to a conventional device, presenting a promising and accurate auto diagnostic AI tool.


Mesenteric Injury in Blunt Abdominal Trauma in Children: Is Early Surgical Intervention Need of the Hour?
There is an increasing trend of observational, nonsurgical management of abdominal injuries in children. We analyzed the feasibility and outcome of our management protocol in managing cases of the mesenteric injuries in blunt trauma abdomen in pediatric age group. Single-center retrospective analysis of pediatric trauma case records from July 2018 to March 2020 was performed. Thirty-four cases of blunt abdominal trauma of whom 13 had mesenteric injuries were reviewed. The male-to-female ratio was 2.2:1, and the mean age was 9.11 ± 4.90 years. Mesentery of the small bowel was the most commonly injured segment and treatment consisted of repair of mesenteric tear, resection, and anastomosis with or without stoma formation. Isolated mesenteric injury of all grades should be managed either with diagnostic laparoscopy or laparotomy irrespective of hemodynamic status.


Less amputations for diabetic foot ulcer from 2008 to 2014, hospital management improved but substantial progress is still possible: A French nationwide study.
To assess the improvement in the management of diabetes and its complications based on the evolution of hospitalisation rates for diabetic foot ulcer (DFU) and lower extremity amputation (LEA) in individuals with diabetes in France. Data were provided by the French national health insurance general scheme from 2008 to 2014. Hospitalisations for DFU and LEA were extracted from the SNIIRAM/SNDS French medical and administrative database. In 2014, 22,347 hospitalisations for DFU and 8,342 hospitalisations for LEA in patients with diabetes were recorded. Between 2008 and 2014, the standardised rate of hospitalisation for DFU raised from 508 to 701/100,000 patients with diabetes. In the same period, the standardised rate of LEA decreased from 301 to 262/100,000 patients with diabetes. The level of amputation tended to become more distal. The proportion of men (69% versus 73%) and the frequency of revascularization procedures (39% versus 46%) increased. In 2013, the one-year mortality rate was 23% after hospitalisation for DFU and 26% after hospitalisation for LEA. For the first time in France, the incidence of a serious complication of diabetes, i.e. amputations, has decreased in relation with a marked improvement in hospital management.


Retention of immunosuppressant activity in an ascomycin analogue lacking a hydrogen-bonding interaction with FKBP12.
C24-Deoxyascomycin was prepared in a two-step process from ascomycin and evaluated for its immunosuppressant activity relative to ascomycin and FK506. An intermediate in the synthetic pathway, Delta(23,24)-dehydroascomycin, was likewise evaluated. Despite lacking the hydrogen-bonding interactions associated with the C24-hydroxyl moiety of ascomycin, C24-deoxyascomycin was found to be equipotent to the parent compound both in its immunosuppressive potency and in its interaction with the immunophilin, FKBP12. Conversely, Delta(23,24)-dehydroascomycin which also lacks the same hydrogen-bonding interactions did not exhibit this potency. NMR studies were conducted on the FKBP12/C24-deoxyascomycin complex in an attempt to understand this phenomenon at the molecular level. The NMR structures of the complexes formed between FKBP12 and ascomcyin or C24-deoxyascomcyin were very similar, suggesting that hydrogen-bonding interactions with the C24 hydroxyl moiety are not important for complex formation.


Acceptability, reliability, and validity of a vaginal insert for the self-assessment of endometriosis-associated deep dyspareunia: a cross-sectional study.
Approximately half of people with endometriosis experience deep dyspareunia; however, there is no means of objective self-testing of endometriosis-associated deep dyspareunia. The aim of this study was to assess the acceptability, test-retest reliability, and validity of a vaginal insert for a self-assessment of endometriosis-associated deep dyspareunia. Participants were recruited from a tertiary endometriosis center. Inclusion criteria were: 19 to 49 years of age, self-reported deep dyspareunia of ≥4 of 10, and surgically confirmed endometriosis. Participants completed 2 self-assessments using the vaginal insert to self-assess tenderness at the right and left pelvic floor, bladder, cervix-uterus, and posterior cul-de-sac (vaginal fornix). The participants recorded tenderness at each pelvic site and completed a questionnaire regarding the acceptability of the vaginal insert to assess deep dyspareunia. Test-retest reliability was assessed by correlating the tenderness scores between the 2 assessment dates. Over a 4-week period, the participants also recorded deep dyspareunia severity at each penetrative vaginal sex encounter. Validity was assessed by correlating vaginal insert tenderness to deep dyspareunia severity, and also to tenderness reported on a prior gynecologic pelvic examination. The main outcome measures were the acceptability index score, tenderness (0-10) at each pelvic site, and prospective deep dyspareunia scores (0-10) over 4 weeks. There were 19 participants (mean age 34 ± 7 years) who completed the study. The majority identified as female (94.7%), heterosexual (89.5%), and white (89.5%). The median acceptability index score was 0.72 (interquartile range, 0.66-0.81). For test-retest reliability, the intraclass correlation coefficients were 0.79 (P = .001) for the left pelvic floor, 0.82 (P < .001) for the right pelvic floor, 0.54 (P = .07) for the bladder, 0.89 (P < .001) for the cervix-uterus, and 0.77 (P = .003) for the cul-de-sac. The correlation between the highest self-assessed mean tenderness in each participant and self-reported deep dyspareunia over 4 weeks was r = 0.32, but correlations for each pelvic site varied significantly. Tenderness at each site on prior gynecologist pelvic exam was associated with higher self-assessed mean tenderness with the vaginal insert in each participant (effect sizes = 0.42-0.88). The vaginal insert is acceptable and reliable for the objective self-assessment of endometriosis-associated deep dyspareunia, with initial evidence of validity. A strength was the inclusion of participants who were avoiding sexual activity and a limitation was the small sample size. Future studies with larger sample sizes are required to further establish the validity of the vaginal insert for the self-assessment of endometriosis-associated deep dyspareunia.


Transition metal-induced dehydrogenative coupling of zinc hydrides.
The reaction of terminal zinc hydrides with palladium(0) species afforded an unprecedented [η2-Zn2Pd] trimetallic complex through the reductive elimination of H2. In contrast, the reaction of zinc hydrides with a nickel(0) reagent gave σ-Zn-H coordination complexes with no direct Zn-Zn bonding. Computational studies indicated that an oxidative addition/reductive elimination process at the palladium center might be responsible for the formation of a covalent Zn-Zn bond.


From mirror-touch synesthesia to models of vicarious experience: A reply to commentaries.
In this reply to the eight commentaries to our article, we discuss three important challenges. First, we discuss the relationship of mirror-touch to other forms of synesthesia. We note that synesthetic experiences are generally not mistaken as veridical but this does not mean that they lack percept-like qualities. We acknowledge that neither Threshold Theory nor Self-Other Theory offer a direct account of other forms of synesthesia, although we discuss how the latter could. Second, we discuss alternative explanations. Notably predictive coding offers a different way of framing our current theory, and extending it to related phenomena. Finally, we discuss how mirror-touch synesthesia may relate to other atypical experiences of body ownership such as the rubber hand illusion, and somatoparaphrenia.


Transcriptional analysis of landmark molecular pathways in lung adenocarcinoma results in a clinically relevant classification with potential therapeutic implications.
Lung adenocarcinoma (LUAD) is a molecularly heterogeneous disease. In addition to genomic alterations, cancer transcriptional profiling can be helpful to tailor cancer treatment and to estimate each patient's outcome. Transcriptional activity levels of 50 molecular pathways were inferred in 4,573 LUAD patients using Gene Set Variation Analysis (GSVA) method. Seven LUAD subtypes were defined and independently validated based on the combined behavior of the studied pathways: AD (adenocarcinoma subtype)1-7. AD1, AD4 and AD5 subtypes were associated with better overall survival. AD1 and AD4 subtypes were enriched in epidermal growth factor receptor (EGFR) mutations, whereas AD2 and AD6 showed higher tumor protein p53 (TP53) alteration frequencies. AD2 and AD6 subtypes correlated with higher genome instability, proliferation-related pathways expression and specific sensitivity to chemotherapy, based on data from LUAD cell lines. LUAD subtypes were able to predict immunotherapy response in addition to CD274 (PD-L1) gene expression and tumor mutational burden (TMB). AD2 and AD4 subtypes were associated with potential resistance and response to immunotherapy, respectively. Thus, analysis of transcriptomic data could improve patient stratification beyond genomics and single biomarkers (i.e., PD-L1, TMB) and may lay the foundation for more personalized treatment avenues, especially in driver-negative LUAD.


University Students' knowledge, attitudes, and practices towards the National Premarital Screening Program of Saudi Arabia.
There is an elevated rate of hereditary hemoglobinopathies in Arab populations. To address this, the Saudi government has implemented a mandatory premarital screening (PMS) program. Nevertheless, reports have shown that 48% of genetically incompatible partners have decided to marry. To address this, more information is needed on people's beliefs on these issues. To investigate the knowledge, attitudes, and practices of Taif University students towards the national PMS program. Using a questionnaire, a cross-sectional study was conducted during April-May 2016 on a random sample of university students in Taif City. It was administered electronically and primarily distributed through social media. Most participants (97.4%) were aware that genes may transmit hereditary diseases and had heard about the PMS program. Most participants who partook in the PMS justified it based on preventing disease transmission to their offspring and ensuring their partner's health. A fair number (82.9%) were willing to change their decision to marry in the case of receiving incompatible results. Moreover, the majority of the participants (91.8%) demanded the implementation of a law that prohibits incompatible marriages. Most Taif University students have a generally positive attitude and good intended practices toward PMS. However, targeted educational programs about the importance of PMS are strongly recommended to eliminate all factors that may affect the success of the PMS program.


Prognostic determinants in duodenal injuries.
A retrospective review of 222 consecutive patients with duodenal injuries admitted to an urban Level 1 Trauma Center who subsequently underwent laparotomy during the period July 1980 to April 2002 was performed in an effort to elucidate factors associated with mortality, infectious morbidity, and length of stay in these patients. Predictably, the patients were predominantly male (92.7%) and young (mean age, 31.6 years). The overall mortality rate was 22.5 per cent, with a mortality rate of 18 per cent seen in the first 48 hours. Penetrating trauma was suffered by 88.3 per cent of the patients. Multivariate analysis revealed the performance of a thoracotomy, initial emergency department (ED) systolic blood pressure (SBP) <90 mm Hg, final operating room (OR) core body temperature less than 35 degrees C, and presence of a splenic injury to be the most important predictors of mortality (all P < 0.05). Mortality in the patients undergoing a resuscitative thoracotomy was 88.9 per cent versus 13.3 per cent in those patients not requiring thoracotomy. An initial SBP in the ED <90 was associated with a 46 per cent mortality rate, as compared with 4 per cent in those patients not in shock. A final OR core body temperature of less than 35 degrees C led to a 60 per cent mortality rate versus 8.3 per cent for warmer patients. Patients with a concomitant splenic injury were noted to have a 62.5 per cent mortality rate; those without had a 19.4 per cent mortality rate. The mean length of stay among survivors greater than 48 hours was 16.0 +/- 24.7 days. Univariate analyses revealed lowest OR core body temperature below 35 degrees C, initial OR SBP <90, presence of infection, >5 transfusions, initial ED SBP <90, final OR core temperature <35 degrees C, colon injury, spleen injury, and an injury severity score (ISS) >25 all to be significantly associated with increased length of stay. Multivariate analysis revealed an initial operating room blood pressure of less than 90 mm Hg systolic, the presence of an infection, and greater than 5 blood transfusions to be the factors most significantly correlated with increased length of stay (all P < 0.02). Of 182 patients surviving 48 hours, 98 (54%) developed an infection. Fifty-seven (31%) patients were noted to have wound-related infections, 92 (51%) patients had nosocomial infections, and 50 (27%) patients had both types. The presence of an abdominal arterial injury, an ISS >25, pancreatic injury, and lowest OR core body temperature <35 degrees C were the factors identified on multivariate analysis most significantly correlated with infectious morbidity (all P < 0.05). This data suggests that early efforts to prevent shock and rapidly control bleeding are the most likely efforts to reduce mortality rates in these patients. Those patients with duodenal injury presenting in shock or requiring a thoracotomy for resuscitation did poorly. Splenic injury was the associated injury found on multivariate analysis to be most closely associated with increased mortality. Early control of bleeding and the prevention of infection provide the most significant opportunity for decreasing length of stay. Infections are common with duodenal injuries, and aggressive surveillance should especially be performed in those patients with an abdominal arterial injury, an ISS >25, pancreatic injury, or lowest OR core body temperature <35 degrees C.


Effects of dimpropyridaz on feeding behavior, locomotivity and biological parameters of Aphis gossypii.
Aphis gossypii is a worldwide agricultural pest insect that has developed resistance to multiple pesticides. Dimpropyridaz is a new chordotonal organ regulator and has been registered for control of sap-sucking insects including A. gossypii. For the aim to effectively apply dimpropyridaz for A. gossypii control, it is necessary to clarify the toxic effects of dimpropyridaz on cotton aphids. In the present study, the effects of dimpropyridaz on feeding behavior, locomotivity and biological parameters of A. gossypii were investigated. The bioassay results showed that dimpropyridaz had good insecticidal activity against A. gossypii, with LC50 as 1.91 mg/L at 72 h post exposure. Moreover, the dimpropyridaz treated A. gossypii showed obvious poisoning symptoms of dehydration and shrivel. Through the gentle-touch experiment and feeding experiment, it was found that dimpropyridaz treatment had significant adverse impacts on the locomotivity and feeding behavior of A. gossypii. Compared with the control group, the coordinated movement ability of the treated A. gossypii attenuated, moreover the feeding behavior of A. gossypii was inhibited. The feeding rate decreased by 62.00%, 64.00% and 71.67% after treatment with 50.33 mg/L dimpropyridaz for 24 h, 48 h and 72 h, respectively. Especially, EPG recordings showed that the number of intracellular stylet puncture and the total duration of phloem sap ingestion and concurrent salivation decreased substantially, while the total duration of non-probing increased after exposure to dimpropyridaz. Furthermore, the treatments with LC10 and LC30 of dimpropyridaz significantly reduced the longevity and fecundity of F0, and led to a decrease of the relative fitness of F0 to 0.48 and 0.32, respectively. The net reproductive rate (R0) and mean generation time (T) of F1 generation were also significantly reduced, moreover the duration of reproduction was significantly shortened. In addition, at 72 h post treatment with LC30 dimpropyridaz, the gene expression levels of JHEH and USP of cotton aphids significantly increased, while the expression of FOXO, INR, EcR and INRS decreased. These results provide basis for clarifying the toxicology of dimpropyridaz to cotton aphids, and also are beneficial for effective control of cotton aphid using dimpropyridaz.


Identification of the cytoplasmic regions of fibroblast growth factor (FGF) receptor 1 which play important roles in induction of neurite outgrowth in PC12 cells by FGF-1.
Fibroblast growth factor 1 (FGF-1) induces neurite outgrowth in PC12 cells. Recently, we have shown that the FGF receptor 1 (FGFR-1) is much more potent than FGFR-3 in induction of neurite outgrowth. To identify the cytoplasmic regions of FGFR-1 that are responsible for the induction of neurite outgrowth in PC12 cells, we took advantage of this difference and prepared receptor chimeras containing different regions of the FGFR-1 introduced into the FGFR-3 protein. The chimeric receptors were introduced into FGF-nonresponsive variant PC12 cells (fnr-PC12 cells), and their ability to mediate FGF-stimulated neurite outgrowth of the cells was assessed. The juxtamembrane (JM) and carboxy-terminal (COOH) regions of FGFR-1 were identified as conferring robust and moderate abilities, respectively, for induction of neurite outgrowth to FGFR-3. Analysis of FGF-stimulated activation of signal transduction revealed that the JM region of FGFR-1 conferred strong and sustained tyrosine phosphorylation of several cellular proteins and activation of MAP kinase. The SNT/FRS2 protein was demonstrated to be one of the cellular substrates preferentially phosphorylated by chimeras containing the JM domain of FGFR-1. SNT/FRS2 links FGF signaling to the MAP kinase pathway. Thus, the ability of FGFR-1 JM domain chimeras to induce strong sustained phosphorylation of this protein would explain the ability of these chimeras to activate MAP kinase and hence neurite outgrowth. The role of the COOH region of FGFR-1 in induction of neurite outgrowth involved the tyrosine residue at amino acid position 764, a site required for phospholipase C gamma binding and activation, whereas the JM region functioned primarily through a non-phosphotyrosine-dependent mechanism. In contrast, assessment of the chimeras in the pre-B lymphoid cell line BaF3 for FGF-1-induced mitogenesis revealed that the JM region did not play a role in this cell type. These data indicate that FGFR signaling can be regulated at the level of intracellular interactions and that signaling pathways for neurite outgrowth and mitogenesis use different regions of the FGFR.


Cholesterol kinetics in control and hypercholesterolemic foxhounds.
Foxhounds were made hypercholesterolemic by thyroidectomy and cholesterol feeding. Tracer cholesterol was administered orally, and i.v. and serum samples obtained sequentially for 30 to 89 days at which time the dogs were euthanized and necropsied. Serum and selected tissues were analyzed for cholesterol radioactivity. Kinetics of cholesterol metabolism were calculated. The hypercholesterolemic dogs had large concentrations of cholesterol in the liver. The only kinetic parameter which was correlated with degree of atherosclerosis was the size of pool 2. Absorption of cholesterol was not different in the hypercholesterolemic from control dogs. Tissue cholesterol disappearance rates suggest a third pool of cholesterol having T 1/2 of greater than 200 days. The bile cholesterol kinetics suggest that cholesterol excreted was derived from the very slow pool. Excretion rate and volume appear to be the factors which are most responsible for the inability of the whole animal to maintain homeostasis in cholesterol concentration when a large amount is consumed.


The design of a novel arthroscopy shaver.
Cases of arthroscopic surgery have increased over the past two decades, and arthroscopic shaver systems have become a commonly used orthopedic tool. Nevertheless, most shavers generally have problems such as the cutting edge is not sharp enough and easy to wear. This paper aims to discuss the structural characteristics of BJKMC's (Bojin◊ Kinetic Medical) novel arthroscopic shaver, the double serrated blade. The product's design and verification process are outlined. BJKMC's articular arthroscopy shaver has a "tube in a tube" structure, comprising a stainless steel outer sleeve and a rotating hollow inner tube. The outer sleeve and inner tube have corresponding suction and cutting windows, and there are serrated teeth on the inner and outer casing. To verify the design rationality, it was compared to Dyonics◊'s equivalent product, the Incisor◊ Plus Blade. The appearance, cutting tool hardness, metal pipe roughness, cutting tool wall thickness, tooth profile, and angle, overall structure, and the key dimensions were examined and compared. Compared with Dyonics◊'s Incisor◊ Plus Blade, BJKMC's Double Serrated Blade had a smoother working surface, harder and thinner blade head. Therefore, BJKMC's product may have satisfactory performance when it comes to surgery.


Contribution of Healthy Chickens to Antimicrobial-Resistant Escherichia coli Associated with Human Extraintestinal Infections in Egypt.
Chickens are considered potential reservoirs for human extraintestinal infections with pathogenic Escherichia coli. However, information about genetic relatedness between E. coli from healthy chickens and human patients is still limited. In this study, clinical samples from patients with extraintestinal infections and healthy broiler chickens were collected from geographically related locations in Egypt during the 2nd half of 2015. The recovered isolates were tested for susceptibility against β-lactam antimicrobials and screened for the presence of extended-spectrum β-lactamases (ESBLs) and virulence genes; clonal and phylotypes were also determined. Forty-eight percent (48/100) and 31.3% (50/160) of human and chicken samples were positive for E. coli, respectively. Although only 4% (2/50) of the chicken isolates were resistant to the tested β-lactams, over 58% of human E. coli isolates (28/48) exhibited resistance to cefotaxime. For β-lactamases, 52.1%, 33.3%, 20.8%, and 6.25% of human E. coli were positive for blaCTX-M, blaTEM, blaOXA, and blaCMY, while blaTEM, blaOXA, and blaCMY were found in 32%, 4%, and 34% of chicken isolates, respectively. Low frequencies of virulence genes within human and chicken E. coli isolates were detected by PCR. The majority of E. coli isolates harboring β-lactam resistance genes from human and chicken sources belonged to phylogroup C and B1, respectively. Using pulsed-field gel electrophoresis (PFGE), some E. coli grouped based upon source; however, most clusters contained isolates from both humans and chickens. The above findings suggest that although no single clone appeared to be circulating among E. coli isolates from human and chicken, some shared characteristics exist among isolates from both sources. Increased study will aid to track the dissemination of β-lactam-resistant E. coli from healthy chickens to humans for implementation of effective intervention strategies.


Diversity and Facing Discrimination in Family Medicine Residencies: A CERA Survey of Program Directors.
Increasing the number of underrepresented minorities in medicine (URM) has the potential to improve access and quality of care and reduce health inequities for diverse populations. Having a diverse workforce in residency programs necessitates structures in place for support, training, and addressing racism and discrimination. This study examines reports of discrimination and training initiatives to increase diversity and address discrimination and unconscious bias in family medicine residency programs nationally. This survey was part of the Council of Academic Family Medicine Educational Research Alliance (CERA) 2018 national survey of family medicine residency program directors. Questions addressed the presence of reported discrimination, residency program training about discrimination and bias, and admissions practices concerning physician workforce diversity. We performed univariate and bivariate analyses on CERA survey response data. We received 272 responses to the diversity survey items within the CERA program director survey from 522 possible residency director respondents, yielding a response rate of 52.1%. The majority of residency programs (78%) offer training for faculty and/or residents in unconscious/implicit bias and systemic/institutional racism. A minority of program directors report discrimination in the residency environment, most often reported by patients (13.2%) and staff (7.2%) and least often by faculty (3.3%), with most common reasons for discrimination noted as language or race/skin color. Most family medicine residency program directors report initiatives to address diversity in the workforce. Research is needed to develop best practices to ensure continued improvement in workforce diversity and racial climate that will enhance the quality of care and access for underserved populations.


Guidelines for extracting biologically relevant context-specific metabolic models using gene expression data.
Genome-scale metabolic models comprehensively describe an organism's metabolism and can be tailored using omics data to model condition-specific physiology. The quality of context-specific models is impacted by (i) choice of algorithm and parameters and (ii) alternate context-specific models that equally explain the -omics data. Here we quantify the influence of alternate optima on microbial and mammalian model extraction using GIMME, iMAT, MBA, and mCADRE. We find that metabolic tasks defining an organism's phenotype must be explicitly and quantitatively protected. The scope of alternate models is strongly influenced by algorithm choice and the topological properties of the parent genome-scale model with fatty acid metabolism and intracellular metabolite transport contributing much to alternate solutions in all models. mCADRE extracted the most reproducible context-specific models and models generated using MBA had the most alternate solutions. There were fewer qualitatively different solutions generated by GIMME in E. coli, but these increased substantially in the mammalian models. Screening ensembles using a receiver operating characteristic plot identified the best-performing models. A comprehensive evaluation of models extracted using combinations of extraction methods and expression thresholds revealed that GIMME generated the best-performing models in E. coli, whereas mCADRE is better suited for complex mammalian models. These findings suggest guidelines for benchmarking -omics integration algorithms and motivate the development of a systematic workflow to enumerate alternate models and extract biologically relevant context-specific models.


Treatment of penetrating hepatic injuries: a retrospective analysis of 50 patients.
The aim of this study was to determine the possibility of non-operative treatment via retrospective analysis of our patients. Fifty patients with penetrating hepatic injuries were examined retrospectively with respect to trauma scores, associated injuries, complications, and mortality parameters. Thirteen injuries were caused by firearms, whereas 37 injuries were caused by stab wounds. Forty-three patients (86%) underwent laparotomy and 7 patients (14%) were monitored by CT and clinical findings (nonoperative group, NO group). The laparotomies were evaluated as non-therapeutic in 11 patients (22%) (NTL group) and therapeutic in 32 patients (64%) (TL group). The morbidity and mortality rates were 40 and 10%, respectively. RTS and ISS scores of the nonsurviving and the surviving patients were significantly different. The rates of major venous, grade IV-V hepatic injuries, and gunshot wounds were significantly higher in the nonsurviving patients when compared to the surviving patients. Major venous and grade IV-V hepatic injuries were the primary factors determining mortality and these injuries generally occurred as a result of firearm injuries. NTL occurring at a rate of 22% would decrease to 2% if 'RTS <7' criteria was added to the hemodynamic instability and/or peritoneal irritation findings determining surgical candidacy.


Appropriate use of fluoroquinolones in children.
With the increasing resistance to antibiotics among common bacterial pathogens, challenges associated with the use of fluoroquinolones (FQs) in paediatrics have emerged. The majority of FQs have favourable pharmacokinetic properties, although these properties can differ in children compared with adults. Moreover, all FQs have broad antimicrobial activity both against Gram-positive and Gram-negative bacteria. However, only some FQs for which adequate studies are available have been approved for use in children in a limited number of clinical situations owing to the supposed risk of development of severe musculoskeletal disorders, as demonstrated in juvenile animals. Recent short- and long-term evaluations appear to indicate that, at least for levofloxacin, this risk, if present at all, is marginal. This marginal risk could lead to more frequent use of FQs in children, even to treat diseases for which several other drugs with documented efficacy, safety and tolerability are considered the first-line antibiotics. However, for most of the FQs, adequate long-term studies of safety are not available. This indicates that the use of FQs should be limited to selected respiratory infections (including tuberculosis), exacerbation of lung disease in cystic fibrosis, central nervous system infections, enteric infections, febrile neutropenia, as well as serious infections attributable to FQ-susceptible pathogen(s) in children with life-threatening allergies to alternative agents. When considering diseases that could benefit from the use of FQs, particular attention must be paid to the choice of drug and its dosage, considering that not all of the FQs have been evaluated in different diseases.


Bilobectomy and amphotericin B in a case of endobronchial mucormycosis.
Lung mucormycosis is a rare fungal infection that has been described mainly in oncologic and diabetic patients. We here report the case of an endobronchial lesion caused by this fungus in an immunocompetent person. Prompt diagnosis, and therapy with Amphotericin B and surgery, permitted the complete resolution.


Eye injuries: a prospective survey of 5671 cases.
5671 patients with injuries presenting to a busy eye casualty department were examined prospectively to determine the incidence, aetiology, and severity of the injury. Of these cases 69.9% occurred at work, 18.3% during leisure and domestic activities (excluding recognised sport), 2.3% during sport, and 1.9% were due to assaults; contact lens injury occurred in a further 2.3%, and the cause was unknown in 5.3%. One hundred and two (1.8%) patients required admission to hospital, and of their injuries 13.7% occurred at work, 22.6% during leisure and domestic activities, 42.2% at sport, 18.6% during assault, and 2.9% from an unknown cause. Children under 10 years of age accounted for 4% of the total patients presenting, but for more than 18% of those admitted and 26% of ocular penetrations. 98.3% of all injuries involved periorbital or superficial ocular structures only. The remainder involved intraocular structures. The majority of serious injuries were contusional. There was only one case of intraocular foreign body. This indicates that, while most injuries still occur in the workplace, sport and leisure activities account for the majority of serious injuries. While most injuries do not threaten sight, they do so disproportionately in children.


Development and validation method for estimation paclitaxel in plasma.
Paclitaxel (PTX) is an anticancer drug used in the treatment of many cancer , alone or in combination with other anti-tumors. It has a narrow therapeutic range, a large inter and intra-individual pharmacokinetic variability and haematological toxicity. The most effective pharmacokinetic parameter seems to be the time during which the plasma concentration is over 0.05 μmol/L. To develop and validate a new method for PTX quantitation in plasma using HPLC with UV/visible detection. A rapid HPLC-UV method was developed for the determination of PTX level in plasma. All solvents used were HPLC grade. After liquid-liquid extraction, chromatographic separation was achieved using an RP 18 (250 mm) column. The mobile phase was composed of acetonitrile and 0.1 M potassium dihyrogenophosphate (49/51) (v/v). Clonazepam was used as internal standard. This technique was linear over the range 50 ng/mL to 1500 ng/mL (r= 0.998). The evaluation of precision showed that our method is repeatable with a within-day coefficient of variation (CV) ranging from 6.94 to 18.78 % and reproducible for three studied concentrations low, medium and high with day-to-day CV of 14.92, 10.46 and 11.8% respectively. Under these conditions, each analysis required no longer than 12.81 min. We have developed and validate a new assay for PTX monitoring using HPLC with UV detection which is sensible, specific, reliable and easy to carry out in clinical use for its therapeutic drug monitoring.


[Localization of crystallins in Muellerian cells in the grass frog retina].
Cell localization of 23 kDa- and 35 kDa-crystallins in the retina of adult common frogs Rana temporaria L. was studied using indirect immunofluorescence. Intense specific fluorescence of both crystallins was observed all over the retina, in both periphery and central area. It was localized in elongated radially oriented cells, whose bodies were located in the inner nuclear layer. These cells gave many fluorescing processes in the same layer and main processes in the outer nuclear and ganglion layers, one in each. The processes formed a strong network of fibers around the photoreceptor and ganglion cells. Intense fluorescence was also observed in the layer of nerve fibers and adjoining inner limiting membrane. The distribution and morphology of crystalline-containing cells mostly coincides with what is known for the Muller cells of vertebrate eye. The identity of the cells we described and Muller cells was also confirmed using the antiserum to glial fibrillary acidic protein.


VIDAS Salmonella (SLM) assay method EasySLM with ChromID Salmonella (SM2) Agar. Performance Tested Method 020901.
A method modification study was conducted for the VIDAS Salmonella (SLM) assay (AOAC Performance Tested Method 020901) using the EasySLM method to validate a matrix extension for peanut butter. The VIDAS EasySLM method is a simple enrichment procedure compared to traditional Salmonella methods, requiring only pre-enrichment and a single selective enrichment media, Salmonella Xpress 2 (SX2) broth. SX2 replaces the two selective broths in traditional methods and eliminates the M broth transfer, incubation, and subsequent pooling of M broths prior to VIDAS assay. The validation study was conducted under the AOAC Research Institute Emergency Response Validation program. VIDAS SLM was compared to the U.S. Food and Drug Administration's Bacteriological Analytical Manual (FDA-BAM) method for detection of S. enterica ser. Typhimurium in peanut butter. All peanut butter samples were prepared, blind-coded, and shipped to the method developers' laboratory by Q Laboratories. In addition, Q Laboratories performed most probable number and reference method analyses on peanut butter samples. The VIDAS EasySLM ChromID Salmonella (SM2) Agar was previously validated in the Performance Tested Methods program for the detection of Salmonella in roast beef, raw ground pork, turkey, pork sausage, raw chicken breast, dry pet food, whole milk, ice cream, bagged spinach, shrimp (raw, peeled), raw cod, spent irrigation water, pecans, peanut butter, dry pasta, cake mix, ground black pepper, nonfat dry milk, liquid eggs, cantaloupe, and orange juice. In the matrix extension study for peanut butter, the VIDAS EasySLM method was shown to be equivalent to the appropriate reference culture procedure using both buffered peptone water pre-enrichment and the FDA-BAM lactose pre-enrichment in the two-step enrichment method with SX2 media. The current study extends the validation to include peanut butter.


Scatter correction based on adaptive photon path-based Monte Carlo simulation method in Multi-GPU platform.
Monte Carlo (MC)-based simulation is the most precise method in scatter correction for Cone-beam CT (CBCT). Nonetheless, the existing MC methods cannot be fully applied in clinical due to its low efficiency. The traditional MC simulations perform calculations via a particle-by-particle scheme, which leads to high computation costs because abundant photons do not reach the X-ray detector in transport. The conventional approaches cannot control where the particle ends. Hence, it unavoidably waste lots of time in transporting numerous photons that have no contribution to the signal at the detector, yielding a low computational efficiency. To solve the problem, an innovative GPU-based Metropolis MC (gMMC) method was proposed. Compared with the traditional ones, the Metropolis based algorithm utilizes a path-by-path sampling method. The method can automatically control each particle path and eventually accelerate the convergence. In this paper, we firstly take planning CT image as prior information because of its precise CT value, and utilize gMMC to estimate scatter signal. Then the scatter signals are removed from the raw CBCT projections. Afterwards, FDK reconstruction is performed to obtain the corrected image,some accelerating strategies including reducing photon history number, pixels sampling, projection angles sampling and reconstructed image down-sampling achieve adaptive fast CBCT image reconstruction. For having high computational efficiency, we implemented the whole workflow on a 4-GPU workstation. In order to verify the feasibility of the the method, the experiment of several cases are conducted including simulation, phantom, and real patient cases. Results indicate that the image contrast becomes better, the scatter artifacts are eliminated. The maximum error (emax), the minimum error (emin), the 95th percentile error (e95%), average error (¯e) are reduced from 264, 56, 14 and 21 HU to 28, 10, 3 and 7 HU in full-fan case, and from 387, 5, 19 and 95 HU to 39, 2, 2 and 6 HU in the half-fan case. In terms of computation time, the MC simulation time of all cases is within 2.5 seconds, and the total time is within 15 seconds.


Mycobacterium avium infection improved by microbial substitution of fungal infection.
We reported a case of Mycobacterium avium infection in which disease activity appeared to have been suppressed after fungal infection. After the increase in β-D-glucan, her symptoms of fever and chest pain disappeared. We think this phenomenon may be microbial substitution and mild fungal infection may improve the activity due to M avium.


Adenosine A1 receptor-mediated inhibition of adenylate cyclase in rabbit retina.
The purine adenosine has been postulated as playing a role in CNS neurotransmission or modulation. Evidence is now provided that inhibition of adenylate cyclase in rabbit retinal homogenates is mediated via adenosine A1 receptors. Nanomolar concentrations of the A1 receptor agonists, cyclohexyladenosine (CHA) and phenylisopropyladenosine (PIA), significantly inhibited the activity of forskolin-stimulated adenylate cyclase in preparations in which endogenous adenosine was destroyed by pretreatment with adenosine deaminase. With increasing concentrations of either agonist, biphasic effects on enzyme activity were observed. The effect of the mixed A1-A2 agonist, N-ethyl-carboxamidoadenosine (NECA), on forskolin-stimulated, as well as basal adenylate cyclase, was also investigated. At micromolar concentrations, NECA significantly increased the activity of adenylate cyclase. Isobutylmethylxanthine, a potent antagonist at extracellular adenosine receptors, blocked the effects observed with PIA, CHA, and NECA. The uptake of both 3H-CHA and 3H-adenosine into retinal cells was demonstrated autoradiographically. Both agonists labeled ganglion cell bodies and certain cell bodies located in the proximal portion of the inner nuclear layer.


Suitability of the Binaural Interaction Component for Interaural Electrode Pairing of Bilateral Cochlear Implants.
Although bilateral cochlear implants (BiCIs) have succeeded in improving the spatial hearing performance of bilateral CI users, the overall performance is still not comparable with normal hearing listeners. Limited success can be partially caused by an interaural mismatch of the place-of-stimulation in each cochlea. Pairing matched interaural CI electrodes and stimulating them with the same frequency band is expected to facilitate binaural functions such as binaural fusion, localization, or spatial release from masking. It has been shown in animal experiments that the magnitude of the binaural interaction component (BIC) derived from the wave-eV decreases for increasing interaural place of stimulation mismatch. This motivated the investigation of the suitability of an electroencephalography-based objective electrode-frequency fitting procedure based on the BIC for BiCI users. A 61 channel monaural and binaural electrically evoked auditory brainstem response (eABR) recording was performed in 7 MED-EL BiCI subjects so far. These BiCI subjects were directly stimulated at 60% dynamic range with 19.9 pulses per second via a research platform provided by the University of Innsbruck (RIB II). The BIC was derived for several interaural electrode pairs by subtracting the response from binaural stimulation from their summed monaural responses. The BIC based pairing results are compared with two psychoacoustic pairing methods: interaural pulse time difference sensitivity and interaural pitch matching. The results for all three methods analyzed as a function of probe electrode allow for determining a matched pair in more than half of the subjects, with a typical accuracy of ± 1 electrode. This includes evidence for statistically significant tuning of the BIC as a function of probe electrode in human subjects. However, results across the three conditions were sometimes not consistent. These discrepancies will be discussed in the light of pitch plasticity versus less plastic brainstem processing.


The significance of neuroglobin in the brain.
Since its discovery in 2000, neuroglobin (Nb) has been demonstrated to have an essential and conserved function in vertebrates with the consequential discovery of a neuroprotective role. Nb is a member of the globin superfamily and is predominantly expressed in neurons of the central and peripheral nervous system. Thorough studies have been performed to elucidate the molecular structure of Nb and its ligand binding characteristics. The precise physiological function and mechanism of action of Nb is beginning to be established, with a number of hypotheses having been put forward. While Nb shares an intrinsic affinity for low-molecular weight diatomic gases similar to other globins, the relatively low level of Nb expression in cerebral neurons places limitations on its potential to function as a reservoir for oxygen, especially during periods of acute ischemia. In vitro studies have suggested that the neuroprotective role of Nb may be due to its ability to scavenge reactive oxygen (ROS) and nitrogen (RNS) species. However other studies have proposed Nb as being part of a signalling chain that transmits the redox state of the cell that is protective against oxidative stress or that inhibits apoptosis. This review is intended to summarize the structural, genomic and functional data on neuroglobin to date, thereby providing perspectives for future research on these molecules that may have substantial biomedical implications.


[Dynamics of the direction of static connections in the electrical activity of the cerebral cortex in the cat].
Data have been obtained on three cats for analysis of statistic connections of electrical activity in the visual, parietal and somatosensory cortical zones during development of evoked potentials to photic signal in these areas, and also in periods previous to photic stimulation and on traces of its action. An increase in shown of the influence of activity in the visual area on that of the somatosensory zone in post-stimulus periods and a sharp increase of two-way connections between these areas against the background of a relative equilibrium of direct and backward connections in trace periods.


Minichromosome maintenance protein 2 is a reliable proliferative marker in breast carcinoma.
This is a study aimed to examine the distribution pattern of a specific minichromosome maintenance protein 2 (MCM2) in benign and malignant breast tissue. We also aim to correlate the frequency of expression of MCM2 with the degree of tumor differentiation. We used immunohistochemistry to examine the distribution and expression pattern of MCM2 on formalin-fixed paraffin-embedded tissue sections of benign (n = 30) and malignant breast tissue (n = 70) (IDC 56, DCIS 4, ILC 2, nonductal 4, mixed type 4). We quantified MCM2 expression by calculating a labeling index, which represents the percentage of epithelial nuclei that stained positively. Immunoreactivity was heterogenous in all the 70 malignant cases examined. Epithelial cells in cycle are most frequent at the tumor periphery. Labeling index of MCM2 was greatest in grade 3 (poorly differentiated) and lowest in grade 1 tumors (well differentiated). Minichromosome maintenance protein 2 expression in breast cancer showed a positive association with histologic grade (P < .05). In all the benign breast tissue examined, no proliferating compartments could be characterized. Minichromosome maintenance protein 2 is a useful proliferative marker of breast carcinoma. The frequency of expression of MCM2 showed an inverse correlation with the degree of tumor differentiation.


Interrelationship of heat and relative humidity in the destruction of Clostridium botulinum type E spores on whitefish chubs.
Heat destruction of types B and E Clostridium botulinum spores on whitefish chubs was observed to be dependent upon the relative humidity (RH) in the chamber in which fish were heated. Experimental conditions were designed to simulate those attainable in commercial fish-smoking plants. Low numbers of type E spores were destroyed with regularity, within 30 min, on fish which were held at an internal temperature of 77 C (170.6 F) in an atmosphere of at least 70% RH. However, an internal temperature of 82 C (179.6 F) and a minimum RH of 70% were required to destroy several hundred thousand type E spores. Quantitative estimates of spore destruction were arrived at with a modified most probable number procedure. Type E spore populations were reduced by 2 to 4 logarithms at 77 C (170.6 F), by 5 to 6 logarithms at 82 C (179.6 F), and by more than 6 logarithms at 88 C (190.4 F) when fish were heated in an atmosphere of 70% RH. A 5 to 6 logarithm reduction of spores was also observed when fish inoculated with type B spores were processed at 82 C (179.6 F) in an atmosphere of 70% RH.


Theoretical aspects of genomic variation screening using DNA microarrays.
We present a theoretical model for typical microarray-based single nucleotide polymorphism (SNP) assay of small genomic DNA amount. We derived the adsorption isotherm expressing the on-array hybridization efficiency in terms of genomic target sequence and concentration, oligonucleotide probe sequence and surface density, hybridization buffer, and temperature. This isotherm correctly describes the surface probe density effects, the sensitivity peak, and the melting temperature depression, and is in accord with published experiments. We discuss optimization of parallel SNP genotyping. Our estimates show that SNP detection at a single temperature in aqueous hybridization buffer is restricted by DNA regions that differ by less than 20% in GC content. We predict that the variety of genotyped SNPs could be substantially extended using an assay design with high probe density and a large fraction of probes hybridized.


Helicobacter pylori-related gastric outlet obstruction: is there a role for medical treatment?
The role of Helicobacter pylori in the pathogenesis of duodenal and gastric ulcer and ulcer recurrence is widely known. Bleeding, perforation, and gastric outlet obstruction represent the most serious, potentially life-threatening complications of ulcer disease. At present, the effect of H. pylori eradication on complicated ulcer disease has not been fully established. Case reports exist on the resolution of gastric outlet obstruction after eradication of H. pylori. We report the first case of H. pylori-related gastric outlet obstruction successfully treated with parenteral antibiotics.


[Topographic mapping of the process of synchronizing moments of sharp changes in the human multichannel EEG].
Techniques of evaluation were studied of the EEG synchronization between different EEG derivations in humans. It was shown that the pattern of cortical cooperativity constructed on the basis of classical EEG crosscorrelation and coherence could be substantially supplemented with estimations of coincidences of the moments of sharp changes (MSC) in multichannel EEG recording (the so called "operational synchronization"). The multichannel EEG was recorded in humans during memory task performance. A non-parametric technique was developed on the basis of the family of Kolmogorov-Smirnov's statistics for revealing the MSC in separate EEG realizations and subsequent spatial-temporal mapping of the MSC coincidences in different derivations of the multichannel EEG. It was shown that such maps markedly change depending on the current cognitive activity of the subjects. An attempt was made to interpret the estimations of the frequency and operational synchronization of multichannel human EEG.


Typical Lung Carcinoids with Metastasis: Potential Role of MicroRNAs in the Regulation of Adaptive Immunity Associated with Disease: a Case Study.
Lung carcinoids are uncommon neuroendocrine tumours. Molecular features of lung carcinoids have been poorly defined. microRNAs (miRNAs) are potent gene expression regulators with important roles in cancer development and progression. However, little is known on the role of miRNAs in the pathogenesis of lung carcinoids. Our goals were to identify commonly deregulated miRNAs in a rare case of lung carcinoid of typical histology with metastasis, as well as map miRNA target genes in pathways potentially associated with disease development and progression. miRNA expression profiles were assessed using the TaqMan Low Density Arrays, which is a platform including 384 miRNAs. miRNA profiles were generated in the tumor and its corresponding lymph node metastasis, compared to reference normal lung tissues. Furthermore, miRNA expression was validated in a separate, publicly available external dataset (n=19 typical lung carcinoids; 2/19 were metastatic tumors, compared to six normal lung tissues, GSE77380). Following this analysis, computational tools were applied for data interpretation. miRTarBase was used to determine miRNA-target genes, followed by ToppGene Suite analysis to identify pathways and biological functions. In addition, the expression of genes targeted by miRNAs was validated in a second, separate external dataset (n=13 tumour samples, GSE35679). GEO2R data analysis tool was used in both validation analyses (miRNAs and genes). We identified 15 commonly significantly downregulated miRNAs (fold change, FC≥2 and p<0.05) in the tumour and its paired metastasis, with further decreasing levels in the metastatic lesion. Downregulation of miR-126-3p and miR-146b-5p was validated in the external dataset GSE77380. In addition, SOX2 and TCF4 genes, targeted by miR-126-3p, were consistently overexpressed in a subset of six typical lung carcinoids from the external dataset GSE35679. Pathways analysis showed that miRNAs miR-126-3p and miR-146b-5p target genes with a role in the regulation of adaptive immune response. Our results contribute to the identification of miRNA expression changes in a typical lung carcinoid and its corresponding lymph node metastasis. Down-regulated levels of miR-126-3p and miR-146b-5p and target gene over-expression could play a role in the progression of this case of primary typical lung carcinoid to regional metastasis. Identified miRNAs and target genes are potential candidates for validation in a larger number of cases.


Automated quantification of the spatial extent of perfusion defects and viability on myocardial contrast echocardiography.
The spatial extent of hypoperfusion or viability is important in the treatment of patients with coronary artery disease. We hypothesized that computerized pixel intensity threshold analysis (PITA) could be used for the automated analysis of perfusion defect size during myocardial contrast echocardiography (MCE). For calibration studies, MCE was performed in 6 dogs undergoing ischemia and reperfusion. Infarct size was determined by PITA, which automatically calculates the percentage of pixels within the myocardium that fail to exceed a predetermined threshold of maximum contrast enhancement. A threshold of 10% of maximum yielded infarct sizes that most closely correlated with those determined by histologic staining. For clinical validation, MCE was performed in 30 patients with acute myocardial infarction before primary percutaneous coronary intervention (PCI) for measurement of risk area; and within 5 days and at 4 weeks after PCI to determine infarct size. The defect size by PITA with a 10% threshold value closely correlated with those measured by expert reader planimetry on background-subtracted color-coded image sets (r = 0.95, P < .001). We conclude that automated analysis of perfusion defect size on MCE is possible by PITA. This technique may be useful for rapid and objective analysis of the extent of ischemia and viability, and for clinical experimentation where accurate and sequential analysis of perfusion defect size is imperative.


Exploring the role of NCCR variation on JC polyomavirus expression from dual reporter minicircles.
JC virus (JCV), a ubiquitous human polyomavirus, can cause fatal progressive multifocal leukoencephalopathy (PML) in immune compromised patients. The viral genome is composed of two conserved coding regions separated by a highly variable non-coding control region (NCCR). We analyzed the NCCR sequence from 10 PML JCV strains and found new mutations. Remarkably, the NCCR f section was mutated in most cases. We therefore explored the importance of this section in JCV expression in renal (HEK293H) and glioblastoma (U-87MG) cell lines, by adapting the emerging technology of DNA minicircles. Using bidirectional fluorescent reporters, we revealed that impaired NCCR-driven late expression in glioblastoma cells was restored by a short deletion overlapping e and f sections. This study evidenced a relevant link between JCV NCCR polymorphism and cell-type dependent expression. The use of DNA minicircles opens new insights for monitoring the impact of NCCR variation.


A dominant adenocarcinoma with multifocal ground glass lesions does not behave as advanced disease.
Invasive lung adenocarcinomas increasingly present with synchronous, multifocal, in situ lesions that appear as ground glass opacities (GGOs). The optimal approach in this circumstance (often nonsmokers) remains unclear. We evaluated a general strategy of anatomic resection of the dominant tumor (DT) and wedge resection of accessible ipsilateral GGOs. This is a retrospective review of 39 patients with suspected multifocal in situ adenocarcinomas and 1 DT in a predominantly Caucasian population. Mean follow-up is 30.7 months. Forty-nine percent of patients had no or minimal smoking history; 21% were Asian. The resected DT was pathologically "bronchioloalveolar carcinoma" (26%), minimally invasive adenocarcinoma (5%), adenocarcinoma with bronchioloalveolar features (41%), or moderate well-differentiated adenocarcinoma (28%). The p stage of the DT was IA in 20, IB in 15, and IIA in 4, with mean diameter of 2.6 cm. Thirty-two patients (82%) underwent anatomic resection of the DT; 7 (18%) underwent wedge resection. The mean number of GGOs present initially was 2.7 (range, 1 to 7) with a 5.2-mm mean diameter. An unresected nodule increased in size during follow-up in only 9 patients (23%). The mean diameter growth among these was 3.2 mm, with mean doubling time of 49 months. New GGOs (range, 1 to 8) developed in 16 patients (41%), all of which remained at 7 mm or less. Distant metastasis developed in 2 patients (5.2%); only 1 patient has required intervention for progression of a GGO. The overall survival is 100%. Patients with limited, multifocal, in situ adenocarcinomas and a clinical N0 DT enjoy prolonged survival with generally anatomic resection of the DT and wedge resection of accessible GGOs. These patients should not be considered to harbor T4 or M1a disease.