Gene prediction and annotation in Penstemon (Plantaginaceae): A workflow for marker development from extremely low-coverage genome sequencing.
Penstemon (Plantaginaceae) is a large and diverse genus endemic to North America. However, determining the phylogenetic relationships among its 280 species has been difficult due to its recent evolutionary radiation. The development of a large, multilocus data set can help to resolve this challenge. • Using both previously sequenced genomic libraries and our own low-coverage whole-genome shotgun sequencing libraries, we used the MAKER2 Annotation Pipeline to identify gene regions for the development of sequencing loci from six extremely low-coverage Penstemon genomes (∼0.005×-0.007×). We also compared this approach to BLAST searches, and conducted analyses to characterize sequence divergence across the species sequenced. • Annotations and gene predictions were successfully added to more than 10,000 contigs for potential use in downstream primer design. Primers were then designed for chloroplast, mitochondrial, and nuclear loci from these annotated sequences. MAKER2 identified longer gene regions in all six Penstemon genomes when compared with BLASTN and BLASTX searches. The average level of sequence divergence among the six species was 7.14%. • Combining bioinformatics tools into a workflow that produces annotations can be useful for creating potential phylogenetic markers from thousands of sequences even when genome coverage is extremely low and reference data are only available from distant relatives. Furthermore, the output from MAKER2 contains information about important gene features, such as exon boundaries, and can be easily integrated with visualization tools to facilitate the process of marker development.


Competitive enzyme immunoassay for bovine growth hormone.
We developed an enzyme immunoassay (EIA) for bovine GH (bGH) which is based on indirect competitive immunoassay in culture medium from a bovine pituitary cell culture. 40 microliters cell culture samples (or bGH standard) and bGH antibody (rabbit anti-bGH) were added to the 96 well microplate coated with secondary antibody (Goat anti-rabbit IgG), and incubated for 24 h at 37 degrees C. Biotin-label bGH was added and incubated further for 24 h at 37 degrees C, and biotinylated bGH was linked with streptoavidin-peroxidase. Substrates for peroxidase were added to the plate and incubated for 1 h at 4 degrees C. The enzyme reaction was stopped with 4N H2SO4, and the absorbency at 450 nm was measured with an ELISA Reader. The coefficients of intra-assay and inter-assay variations were 4.13 approximately 7.59% and 3.71 approximately 8.27%, respectively. The regression equation and correlation coefficients with the radioimmunoassay (RIA) were y(RIA) = 1.9986 x (EIA) - 1.3921 and 0.9701 (n = 27), respectively. Collectively, the present assay provides a reliable alternative to RIA and offers the major advantage of eliminating radioactive reagents and counting equipment.


Intranasal calcitonin for prevention and treatment of osteoporosis.
Age-related diseases constitute an ever increasing problem in an ageing population. The objectives of future medical research in this field should not only seek to increase longevity, but also to improve the quality of life of the elderly and to enable them to live independently for a longer period of time. Osteoporosis is a prime example of such a disease, because we have the tool to prevent it, i.e. oestrogen and calcitonin, and because in all probability it will some day be curable.


Attenuation of X and Gamma Rays in Personal Radiation Shielding Protective Clothing.
A collection of personal radiation shielding protective clothing, suitable for use in case of accidents in nuclear facilities or radiological emergency situations involving radioactive agents, was gathered and tested at the Nuclear Protection Department of the National Institute for Nuclear, Chemical and Biological Protection, Czech Republic. Attenuating qualities of shielding layers in individual protective clothing were tested via spectra measurement of x and gamma rays, penetrating them. The rays originated from different radionuclide point sources, the gamma ray energies of which cover a broad energy range. The spectra were measured by handheld spectrometers, both scintillation and High Purity Germanium. Different narrow beam geometries were adjusted using a special testing bench and a set of various collimators. The main experimentally determined quantity for individual samples of personal radiation shielding protective clothing was x and gamma rays attenuation for significant energies of the spectra. The attenuation was assessed comparing net peak areas (after background subtraction) in spectra, where a tested sample was placed between the source and the detector, and corresponding net peak areas in spectra, measured without the sample. Mass attenuation coefficients, which describe attenuating qualities of shielding layers materials in individual samples, together with corresponding lead equivalents, were determined as well. Experimentally assessed mass attenuation coefficients of the samples were compared to the referred ones for individual heavy metals.


Role of cyclic nucleotide signaling in oocyte maturation.
The development of the ovarian follicle, oocyte maturation, and ovulation require a complex set of endocrine, paracrine, and autocrine inputs that are translated into the regulation of cyclic nucleotide levels. Changes in intracellular cAMP mediate the gonadotropin regulation of granulosa and theca cell functions. Likewise, a decrease in cAMP concentration in the oocyte has been associated with the resumption of meiosis. Using pharmacological and molecular approaches, we determined that the expression of cyclic nucleotide phosphodiesterases (PDEs), the enzymes that degrade and inactivate cAMP, is compartmentalized in the ovarian follicle of all species studied, with PDE3 present in the oocytes and PDE4s in granulosa cells. The PDE3 expressed in the mouse oocyte was cloned, and the protein expressed in a heterologous system had properties similar to those of a PDE3A derived from somatic cells. Inhibition of the oocyte PDE3 completely blocked oocyte maturation in vitro and in vivo, demonstrating that the activity of this enzyme is essential for oocyte maturation. Heterologous expression of PDE3A in Xenopus oocyte causes morphological changes distinctive of resumption of meiosis (GVBD), as well as activation of mos translation and MAPK phosphorylation. Using mRNA and antibody microinjection in the Xenopus eggs, we have shown that PDE3 is downstream from the kinase PKB/Akt in the pathway that mediates IGF-1 but not progesterone-induced meiotic resumption. The presence of a similar regulatory module in mammalian oocytes is inferred by pharmacological studies with PDE3 inhibitors and measurement of PDE activity. Thus, PDE3 plays an essential role in the signaling pathway that controls resumption of meiosis in amphibians and mammals. Understanding the regulation of this enzyme may shed some light on the signals that trigger oocyte maturation.


ZCAM, a colour appearance model based on a high dynamic range uniform colour space.
A colour appearance model based on a uniform colour space is proposed. The proposed colour appearance model, ZCAM, comprises of comparatively simple mathematical equations, and plausibly agrees with the psychophysical phenomenon of colour appearance perception. ZCAM consists of ten colour appearance attributes including brightness, lightness, colourfulness, chroma, hue angle, hue composition, saturation, vividness, blackness, and whiteness. Despite its relatively simpler mathematical structure, ZCAM performed at least similar to the CIE standard colour appearance model CIECAM02 and its revision, CAM16, in predicting a range of reliable experimental data.


Octopine dehydrogenase from Pecten maximus: steady-state mechanism.
The steady-state kinetic mechanism of the reaction catalyzed by octopine dehydrogenase [N2-(1-carboxyethyl)-L-arginine:NAD+ oxidoreductase] was investigated at pH 6.9 and 9.2 by studies of substrate inhibition, analogue inhibition, and product inhibition. In the direction of octopine synthesis, the inhibition patterns in the presence of delta- guanidinovalerate and propionate show that NADH binds to the enzyme first followed by L-arginine and pyruvate which bind randomly. In the direction of octopine oxidation, the substrate patterns show that NAD binds to the enzyme before octopine in a rapid equilibrium fashion, and the product inhibition patterns show that the products L-arginine and pyruvate are released in a random fashion. Double, synergistic, substrate inhibition by L-arginine and pyruvate was shown to be due to binding (hypothetically of the imine) to the free enzyme and the enzyme-NAD complex. Furthermore, an alternate minor pathway was demonstrated which includes an enzyme-NADH-octopine complex and an enzyme-octopine complex.


The effects of yeast feed supplementation on turkey performance and pathogen colonization in a transport stress/Escherichia coli challenge.
A commercial yeast culture feed supplement (YC; Celmanax SCP, Vi-COR, Mason City, IA) was provided to turkeys throughout a 16-wk grow-out to determine if it would prevent the effects of stress on production and pathogen colonization. The YC was provided either continuously at 100 g/t (YC-CS) or intermittently during times of stress at 200 g/t (YC-IS). Birds were stressed with an environmental challenge of Escherichia coli and by transporting them in a vehicle for 3 h after which they were penned in new social groups, without feed or water, for an additional 9 h. Turkeys were transported and challenged at 6, 12, and 16 wk of age to model the movement of birds within a 3-stage housing system. The YC-IS was provided only for the first week after hatch and for a 1-wk period encompassing each challenge. At wk 7 and 9, a decrease in BW of challenged birds was prevented by YC-IS but not YC-CS. There were no significant differences in BW due to either challenge or YC during wk 11 and 13. At wk 16, the challenge decreased BW, but there was no improvement in either of the YC treatments. Overall feed conversion ratio (FCR) was increased by transport/E. coli (P < 0.0001). The YC-CS improved FCR of challenged birds by 21 points, whereas YC-IS improved FCR by 36 points and this effect was significant (P = 0.013). The YC-CS tended to decrease both Salmonella and Campylobacter isolation from the ceca of stressed birds (P > 0.05). The YC-IS also tended to decrease Salmonella isolation (P > 0.05) with no effect on Campylobacter isolation. These data suggest that the practice of transporting turkeys decreases performance and that YC-IS may be more effective than YC-CS for alleviating the effects of this stressor on feed efficiency.


Summer exposure assessment of Cu and Zn in drinking water in Shanghai, China.
Daily exposures to chemicals in drinking water are of importance in risk assessment to human health. However, exposure assessment of chemicals in drinking water has seldom been performed in China. Until now, no such a detailed exposure assessment was reported in China. The aim of the present paper is to describe the scenes of daily exposure to Cu and Zn in drinking water by the population of Shanghai, China in the summer of 2003. A total of 163 volunteers and/or homes were selected to be statistically representative of the population of Shanghai, China. The Modified Consumption Habit Exposure Model (CHEM) method combined with the measurement of metals concentrations in stagnant water and actual ingested water was used to describe the daily exposure. The Consumption Habit Exposure Model method results showed that water consumption ranged from 1.7 to 2.2 L/day, varied with different age, gender and work location groups. Chemical results showed that the average Cu and Zn concentrations in stagnant water were much higher than those in actual ingested water (13.5 microg/L and 1.55 mg/L vs. 10.8 microg/L and 0.29 mg/L). Exposure assessment results also showed that volunteers had the highest exposure from the time of after breakfast to before dinner, and had the lowest exposure during the 15 minutes before sleeping. It was also showed that there were few volunteers directly exposed to stagnant water, although six hours stagnant time can be reached in most of the homes in Shanghai in the morning. The mean daily exposures to Cu and Zn were 0.36 microg/kg/day and 0.01 mg/kg/day. Daily intakes of Cu and Zn from drinking water by Shanghai people were 1.8% and 4.4% of the World Health Organization recommendations for minimum total intakes of Cu and Zn for adults, respectively.


Cyclin-dependent kinase-mediated phosphorylation of RBP1 and pRb promotes their dissociation to mediate release of the SAP30·mSin3·HDAC transcriptional repressor complex.
Eukaryotic cell cycle progression is mediated by phosphorylation of protein substrates by cyclin-dependent kinases (CDKs). A critical substrate of CDKs is the product of the retinoblastoma tumor suppressor gene, pRb, which inhibits G(1)-S phase cell cycle progression by binding and repressing E2F transcription factors. CDK-mediated phosphorylation of pRb alleviates this inhibitory effect to promote G(1)-S phase cell cycle progression. pRb represses transcription by binding to the E2F transactivation domain and recruiting the mSin3·histone deacetylase (HDAC) transcriptional repressor complex via the retinoblastoma-binding protein 1 (RBP1). RBP1 binds to the pocket region of pRb via an LXCXE motif and to the SAP30 subunit of the mSin3·HDAC complex and, thus, acts as a bridging protein in this multisubunit complex. In the present study we identified RBP1 as a novel CDK substrate. RBP1 is phosphorylated by CDK2 on serines 864 and 1007, which are N- and C-terminal to the LXCXE motif, respectively. CDK2-mediated phosphorylation of RBP1 or pRb destabilizes their interaction in vitro, with concurrent phosphorylation of both proteins leading to their dissociation. Consistent with these findings, RBP1 phosphorylation is increased during progression from G(1) into S-phase, with a concurrent decrease in its association with pRb in MCF-7 breast cancer cells. These studies provide new mechanistic insights into CDK-mediated regulation of the pRb tumor suppressor during cell cycle progression, demonstrating that CDK-mediated phosphorylation of both RBP1 and pRb induces their dissociation to mediate release of the mSin3·HDAC transcriptional repressor complex from pRb to alleviate transcriptional repression of E2F.


An ab initio and DFT study of some halogen atom transfer reactions from alkyl groups to acyl radical.
Ab initio calculations using the 6-311G**, cc-pVDZ, and (valence) double-zeta pseudopotential (DZP) basis sets, with (MP2, QCISD, CCSD(T)) and without (HF) the inclusion of electron correlation, and density functional (BHandHLYP, B3LYP) calculations predict that the transition states for the reaction of acetyl radical with several alkyl halides adopt an almost collinear arrangement of attacking and leaving radicals at the halogen atom. Energy barriers (DeltaE(double dagger)) for these halogen transfer reactions of between 89.2 (chlorine transfer from methyl group) and 25.3 kJ mol(-1) (iodine transfer from tert-butyl group) are calculated at the BHandHLYP/DZP level of theory. While the difference in forward and reverse energy barriers for iodine transfer to acetyl radical is predicted to be 15.1 kJ mol(-1) for primary alkyl iodide, these values are calculated to be 6.7 and -4.2 kJ mol(-1) for secondary and tertiary alkyl iodide respectively. These data are in good agreement with available experimental data in that atom transfer radical carbonylation reactions are sluggish with primary alkyl iodides, but proceed smoothly with secondary and tertiary alkyl iodides. These calculations also predict that bromine transfer reactions involving acyl radical are also feasible at moderately high temperature.


[50th Anniversary of Psychiatric Attitude Research in Germany].
Objective Analysis of the development of psychiatric attitude research in Germany over the past 50 years. Methods Historical overview based on systematic reviews of population-based studies. Results In the beginning, public attitudes towards people with mental disorders and towards psychiatry were studied from a sociological perspective, later on research was guided by a social psychological or individual psychological approach. This provided important insights into processes underlying the stigma of mental illness (e. g., role of labeling, causal attributions, beliefs into a continuum between mental health and illness, prevalence estimates etc.). Substantial contributions have also been made as regards the study of time trends in public attitudes and beliefs about mental illness, the investigation of the impact of the media on public attitudes, and the evaluation of the effectiveness of anti-stigma and awareness programs. Conclusion Despite these undisputable advancements it is about time to compliment the individual by a societal perspective. This appears particularly necessary in view of current trends in Germany towards sociopolitical radicalization.


[Perioperative analgesia with continuous peripheral nerve blocks in children].
Recently, regional anaesthesia in children has generated increasing interest. But single injection techniques have a limited duration of postoperative analgesia. Then, continuous peripheral nerve blocks have taken an important position in the anaesthetic arsenal, allowing an effective, safe and prolonged postoperative pain management. As adults, indications for continuous peripheral nerve blocks depend on the analysis of individual benefits/risks ratio. Main indications are intense postoperative pain surgical procedures, with or without postoperative rehabilitation, and complex regional pain syndrome. Contraindications to these procedures are rather similar to those in adults, plus parental and/or children refusal. Continuous peripheral nerve blocks are usually performed under general anaesthesia or sedation in children, and require appropriate equipment in order to decrease the risk of nerve injury. New techniques, such as transcutaneous nerve stimulation or ultrasound guidance, appeared to facilitate nerve and plexus approach identification in paediatric patients. Nevertheless, continuous peripheral nerve block may theoretically mask a compartment syndrome after trauma surgical procedures. Finally, ropivacaine appears to be the most appropriate drug for continuous peripheral nerve blocks in children, requiring low flow rates and concentrations of local anaesthetic. These techniques may facilitate early ambulation by an improved pain management or even postoperative analgesia at home with disposable pumps. One might infer from the current review that excellent pain relief coupled with a reduction of side effects would contribute to improve the quality of life and to decrease the frequency of disabling behavioural modifications in children, sometimes psychologically injured by hospital stay and postoperative pain.


Web-based medical facilitators in medical tourism: the third party in decision-making.
The emergence of web-based medical tourism facilitators (MTFs) has added a new dimension to the phenomenon of cross-border travel. These facilitators are crucial connectors between foreign patients and host countries. They help patients navigate countries, doctors and specialties. However, little attention has been paid to the authenticity of information displayed on the facilitators' web portals, and whether they follow ethical guidelines and standards. This paper analyses the available information on MTF portals from an ethics perspective. It compares 208 facilitators across 47 countries for the services offered. Data were collected from the databases of the Medical Tourism Association and World Medical Resources. India was the most common destination country linked to 81 facilitators. The five countries with the maximum number of facilitators were the USA, the UK, India, Canada and Poland. This paper identifies concerns regarding the information displayed about patients' safety, and the maintenance of confidentiality. There is a need to develop ethical standards for this field.


Fabrication and dispersion evaluation of single-wall carbon nanotubes produced by FH-arc discharge method.
In this work, we produced SWNTs by a hydrogen DC arc discharge with evaporation of carbon anode containing 1 at% Fe catalyst in H2-Ar mixture gas. This was named as FH-arc discharge method. The as-grown SWNTs synthesized by FH-arc discharge method have high crystallinity. An oxidation purification process of as-grown SWNTs with H2O2 has been developed to remove the coexisting Fe catalyst nanoparticles. As a result, SWNTs with purity higher than 90 at% have been achieved. To exhibit remarkable characteristics, CNTs should be separated from the bundles and kept in homogeneous and stable suspensions. For this purpose, the SWNTs prepared by FH-arc discharge method also have been treated by Nanomizer process with some surfactants. SPM images showed that the SWNTs bundles had become thinner and shorter.


Nodular amyloidosis in a patient with systemic scleroderma.
Primary cutaneous amyloidosis may be characterized as macular amyloidosis, lichenoid amyloidosis, or nodular amyloidosis. Nodular amyloidosis results from the deposition of immunoglobulin light chains and may rarely be associated with systemic amyloidosis. We report an unusual case of a patient with systemic scleroderma who developed primary cutaneous nodular amyloidosis on the left lower leg. The diagnosis was confirmed with a skin biopsy with Congo red staining and a novel technique using a laser microdissection and mass spectrometry-based proteomic analysis method for amyloid protein characterization. A work-up for systemic amyloidosis was negative and the patient improved symptomatically with wound care. Patients with primary cutaneous nodular amyloidosis should be followed clinically over time for the possible development of systemic amyloidosis, although the risk of disease progression is likely low.


Relationship of reproductive hormones and neuromuscular disease of the gastrointestinal tract.
Functional disorders of the gastrointestinal tract comprise a common but ill-defined group of diseases; they primarily afflict women. Although predominantly involving nerve and muscle, the cellular and molecular bases of the pathogenesis of these functional disorders are unknown. Clinical studies indicate that some result from neural dysfunction within the enteric nervous system, others may be due to muscular problems, and the causes of still others remain unknown. Laboratory studies have shown that ovarian products such as progesterone, luteinizing hormone, human chorionic gonadotropin, and relaxin (but not estrogen), are neural antagonists of gastrointestinal motility. The production and secretion of these ovarian substances are controlled by gonadotropin-releasing hormone (GnRH) released from the hypothalamus; they probably act on gamma-aminobutyric acid receptors and alter chloride influx into the cell. GnRH analogs are effective drugs that downmodulate the hypothalamic-pituitary-gonadal axis and inhibit the secretion of gonadal products involved in such hormone-dependent diseases as endometriosis and prostate cancer. Acting on the GnRH receptors (seven transmembrane domain receptors) on myenteric neurons, GnRH analogs are also effective neural modulators in such disorders as functional bowel disease. These analogs are a promising new group of compounds that may be used to treat difficult gastrointestinal problems.


FIN219/JAR1 and cryptochrome1 antagonize each other to modulate photomorphogenesis under blue light in Arabidopsis.
Plant development is affected by the integration of light and phytohormones, including jasmonates (JAs). To address the molecular mechanisms of possible interactions between blue light and JA signaling in Arabidopsis thaliana, we used molecular and transgenic approaches to understand the regulatory relationships between FAR-RED INSENSITIVE 219 (FIN219)/JASMONATE RESISTANT1 (JAR1) and the blue-light photoreceptor cryptochrome1 (CRY1). FIN219 overexpression in the wild type resulted in a short-hypocotyl phenotype under blue light. However, FIN219 overexpression in cry1, cry2 and cry1cry2 double mutant backgrounds resulted in phenotypes similar to their respective mutant backgrounds, which suggests that FIN219 function may require blue light photoreceptors. Intriguingly, FIN219 overexpression in transgenic plants harboring ectopic expression of the C terminus of CRY1 (GUS-CCT1), which exhibits a hypersensitive short-hypocotyl phenotype in all light conditions including darkness, led to a rescued phenotype under all light conditions except red light. Further expression studies showed mutual suppression between FIN219 and CRY1 under blue light. Strikingly, FIN219 overexpression in GUS-CCT1 transgenic lines (FIN219-OE/GUS-CCT1) abolished GUS-CCT1 fusion protein under blue light, whereas GUS-CCT1 fusion protein was stable in the fin219-2 mutant background (fin219-2/GUS-CCT1). Moreover, FIN219 strongly interacted with COP1 under blue light, and methyl JA (MeJA) treatment enhanced the interaction between FIN219 and GUS-CCT1 under blue light. Furthermore, FIN219 level affected GUS-CCT1 seedling responses such as anthocyanin accumulation and bacterial resistance under various light conditions and MeJA treatment. Thus, FIN219/JAR1 and CRY1 antagonize each other to modulate photomorphogenic development of seedlings and stress responses in Arabidopsis.


Evaluation of the Antioxidant and Antiangiogenic Activity of a Pomegranate Extract in BPH-1 Prostate Epithelial Cells.
Benign prostatic hypertrophy (BPH) is a noncancerous enlargement of the prostate gland that develops from hyper-proliferation of the stromal and epithelium region. Activation of pathways involving inflammation and oxidative stress can contribute to cell proliferation in BPH and tumorigenesis. Agricultural-waste-derived extracts have drawn the attention of researchers as they represent a valid and sustainable way to exploit waste production. Indeed, such extracts are rich in bioactive compounds and can provide health-promoting effects. In particular, extracts obtained from pomegranate wastes and by-products have been shown to exert antioxidant and anti-inflammatory effects. This study focused on the evaluation of the anti-angiogenic effects and chemopreventive action of a pomegranate extract (PWE) in cellular models of BPH. In our experimental conditions, we observed that PWE was able to significantly (p < 0.001) reduce the proliferation and migration rates (up to 60%), together with the clonogenic capacity of BPH-1 cells concomitantly with the reduction in inflammatory cytokines (e.g., IL-6, PGE2) and pro-angiogenic factor (VEGF-ADMA) release. Additionally, we demonstrated the ability of PWE in reducing angiogenesis in an in vitro model of BPH consisting in transferring BPH-1-cell-conditioned media to human endothelial H5V cells. Indeed, PWE was able to reduce tube formation in H5V cells through VEGF level reduction even at low concentrations. Overall, we confirmed that inhibition of angiogenesis may be an alternative therapeutic option to prevent neovascularization in prostate tissue with BPH and its transformation into malignant prostate cancer.


Cell density-dependent proliferation in frequently-fed peripheral blood mononuclear cell cultures.
Our goal was to produce granulocyte progenitor (CFU-G) and post-progenitor (CD15(+)CD11b(+/-)) cells for subsequent transplantation. We hypothesized that increasing the feeding frequency and maintaining constant densities may overcome inhibitory growth conditions (i.e. low pH) in high-density cultures. To study the effect of cell density on total cell expansion, differentiation and lactate production, 50% daily medium exchanges were used in cultures of peripheral blood mononuclear cells (PB MNC) maintained at constant densities (ranging from 5 x 10(4)cells/mL to 2.5 x 10(6)cells/mL). We observed a significant increase in total cell expansion when the density was increased from 5 x 10(4) cells/mL to 1 x 10(6) cells/mL, but a further increase to 2.5 x 10(6)cells/mL resulted in a decline in cell expansion. Increasing feeding to 90% daily exchange in cultures with 2.5 x 10(6) cells/mL did not enhance cell expansion; nor did reducing the extent of feeding in cultures with 5 x 10(4) cells/mL to 10% daily exchange. We did not observe a relationship between cell density and the percentage of granulocyte progenitor and post-progenitor (CD15(+)CD11b(-/+)) cells. While specific lactate production (q(lac)) in cultures with 2.5 x 10(6) cells/mL was approximately 60% of those observed in lower density cultures by Day 13, this difference was largely eliminated by increasing the extent of feeding in cultures with 2.5 x 10(6) cells/mL. Our results suggest that feeding rates must be adjusted according to cell density to maximize culture performance. They also suggest that cellular crowding on the culture surface can limit expansion in suspension (nonadherent) cultures.


Self-assembled monolayers and polymer brushes in biotechnology: current applications and future perspectives.
The chemistry and topography of a surface affect biological response and are of fundamental importance, especially when living systems encounter synthetic surfaces. Most biomolecules have immense recognition power (specific binding) and simultaneously have a tendency to physically adsorb onto a solid substrate without specific receptor recognition (nonspecific adsorption). Therefore, to create useful materials for many biotechnology applications, interfaces are required that have both enhanced specific binding and reduced nonspecific binding. Thus, in applications such as sensors, the tailoring of surface chemistry and the use of micro or nanofabrication techniques becomes an important avenue for the production of surfaces with specific binding properties and minimal background interference. Both self-assembled monolayers (SAMs) and polymer brushes have attracted considerable attention as surface-active materials. In this review, we discuss both of these materials with their potential applications in biotechnology. We also summarize lithographic methods for pattern formation using combined top-down and bottom-up approaches and briefly discuss the future of these materials by describing emerging new applications.


Oxidation of lysozyme induced by peroxyl radicals involves amino acid modifications, loss of activity, and formation of specific crosslinks.
The present work examined the oxidation and crosslinking of the anti-bacterial enzyme lysozyme (Lyso), which is present in multiple biological fluids, and released from the cytoplasmic granules of macrophages and neutrophils at sites of infection and inflammation. It is therefore widely exposed to oxidants including peroxyl radicals (ROO•). We hypothesized that exposure to ROO• would generate specific modifications and inter- and intra-protein crosslinks via radical-radical reactions. Lyso was incubated with AAPH (2,2'-azobis(2-methylpropionamidine) dihydrochloride) as a ROO• source. Enzymatic activity was assessed, while oxidative modifications were detected and quantified using electrophoresis and liquid chromatography (UPLC) with fluorescence or mass detection (MS). Computational models of AAPH-Lyso interactions were developed. Exposure of Lyso to AAPH (10 and 100 mM for 3 h, and 20 mM for 1 h), at 37 °C, decreased enzymatic activity. 20 mM AAPH showed the highest efficiency of Lyso inactivation (1.78 mol of Lyso inactivated per ROO•). Conversion of Met to its sulfoxide, and to a lesser extent, Tyr oxidation to 3,4-dihydroxyphenylalanine and diTyr, were detected by UPLC-MS. Extensive transformation of Trp, involving short chain reactions, to kynurenine, oxindole, hydroxytryptophan, hydroperoxides or di-alcohols, and N-formyl-kynurenine was detected, with Trp62, Trp63 and Trp108 the most affected residues. Interactions of AAPH inside the negatively-charged catalytic pocket of Lyso, with Trp108, Asp52, and Glu35, suggest that Trp108 oxidation mediates, at least partly, Lyso inactivation. Crosslinks between Tyr20-Tyr23 (intra-molecular), and Trp62-Tyr23 (inter-molecular), were detected with both proximity (Tyr20-Tyr23), and chain flexibility (Trp62) appearing to favor the formation of covalent crosslinks.


Effect of cholinergic crisis on the potency of different emergency anaesthesia protocols in soman-poisoned rats.
In a military or terrorist scenario, combination of organophosphorus compounds (OP) poisoning with physical trauma requiring surgical treatment and thus general anaesthesia are possible. Previous in vitro studies showed an altered potency of relevant anaesthetics during cholinergic crisis. Hence, it is not clear, which anaesthetics are suitable to achieve the necessary stage of surgical anaesthesia in OP poisoning. In the present study, different anaesthetic regimens (ketamine-midazolam, propofol-fentanyl, thiopental-fentanyl), relevant in military emergency medicine, were examined in soman-poisoned rats. Clinical signs and cardiovascular variables were recorded continuously. Blood samples for acetylcholinesterase (AChE) activity were drawn. After euthanasia or death of the animals, brain and diaphragm were collected for cholinesterase assays. Propofol-fentanyl and thiopental-fentanyl resulted in surgical anaesthesia throughout the experiments. With ketamine-midazolam, surgical anaesthesia without respiratory impairment could not be achieved in pilot experiments (no soman challenge) and was therefore not included in the study. Soman-poisoned and control animals required a comparable amount of propofol-fentanyl or thiopental-fentanyl. In combination with atropine, significantly less propofol was needed. Survival rate was higher with thiopental compared to propofol. Atropine improved survival in both groups. Blood and tissue AChE activities were strongly inhibited after soman administration with and without atropine treatment. The current in vivo study did not confirm concerns of altered potency of existing anaesthetic protocols for the application of propofol or thiopental with fentanyl due to soman poisoning. Despite severe cholinergic crisis, sufficient anaesthetic depth could be achieved in all animals. Further experiments in in vivo models closer to human pharmaco- and toxicokinetics (e.g., swine) are required for confirmation of the initial findings and for improving extrapolation to humans.


Maxillary implant overdentures retained with bars or solitary attachments: A 5-year randomised controlled trial.
Purpose To compare the 5-year follow-up outcomes of maxillary overdentures retained by bars or solitary attachments.Methods Fifty consecutively selected fully edentulous patients experiencing problems with their conventional denture received four implants and were randomly allocated to receive a maxillary overdenture with either bar- or solitary attachment retention. Marginal bone level change (primary outcome), implant- and overdenture survival rate, clinical- and patient related outcome measures were recorded at baseline, and after 1 and 5 years. Biological and technical complications were recorded throughout the entire follow-up period.Results After 5 years, the mean marginal bone level change was higher in the solitary attachment group (-1.41±1.38mm, P=0.024) than in the bar group (-0.99±0.96mm). Also, fewer implants survived in the solitary attachment group (89.5%, P=0.027) than in the bar group (96.3%). The overdenture survival rate was 95.0% and 91.3% in the bar and solitary attachment group, respectively. Although the clinical and patient related outcomes were favourable and did not differ significantly between the groups, the peri-implantitis incidence was 25.8% in the solitary attachment group and 5.1% in the bar group. Any technical complications were minor.Conclusions In maxillary 4-implant overdenture therapy, the marginal bone level, implant survival rate and the number of complications are better with bar attachments than with solitary attachments. Both groups' clinical and patient related outcome measure scores were equal throughout the entire follow-up period.


Molecular identification of archaic bones as a native Korean black bear: implications for the ongoing bear restoration program.
The genetic investigation of the archeological or museum samples, including endangered species, provides vital information necessary to plan, implement, and revisit conservation strategies. In South Korea, the Asian black bear went almost extinct in wild by 2002, without leaving any authentic specimens representing the native population. Recently researchers found a set of animal bones in a natural cave in Mt. Taebaek (South Korea), suspected to be of a bear. In the present study, we undertook a molecular investigation and radiocarbon dating to establish the species' identity, phylogenetic position, and approximate age of the recovered specimen. The genetic investigation (CytB, COI, D-loop, SRY, and ZFX-ZFY) identified the sample as a male Asian black bear with close phylogenetic affinity with Northeast Asian bears. Radiocarbon dating estimated the bones to be aged 1800-1942 calAD. These findings indicate that the bone specimens found in the natural cave in Mt. Taebaek were from an individual that naturally inhabited South Korea long before the importing of farm bears (the 1980s) and initiation of wild population restoration (2004). The present study provides the first genetic information record of the native South Korean black bear. Our findings reaffirm the appropriateness of the ongoing bear restoration program in South Korea, with the reintroduction of individuals from North Korea and Russia.


Beta-blocker therapy in the Cardiac Arrhythmia Suppression Trial. CAST Investigators.
The Cardiac Arrhythmia Suppression Trial (CAST) showed antiarrhythmic drug suppression of asymptomatic or mildly symptomatic ventricular arrhythmias in survivors of myocardial infarction to be harmful. This study retrospectively searched the CAST results for evidence of mortality and morbidity reduction in patients receiving optional beta-blocker therapy. All enrolled (n = 2,611) and suppressed main study (n = 1,735) CAST patients with an ejection fraction of < or = 40% were examined using univariate analysis, Kaplan-Meier curves, and a Cox proportional-hazards multivariate analysis with respect to optional beta-blocker therapy prescribed at baseline. CAST patients receiving beta-blocker therapy had significantly enhanced survival at 30 days, and at 1 and 2 years of follow-up against all-cause and arrhythmic death or nonfatal cardiac arrest. Multivariate analysis showed beta-blocker therapy to be independently associated with a one-third reduction in arrhythmic death or cardiac arrest (p = 0.036). In CAST patients with a history of congestive heart failure, beta-blocker therapy was independently associated with longer time to occurrence of new or worsened congestive heart failure (p = 0.015). This study supports the secondary preventive benefit of beta-blocker therapy in high-risk post-myocardial infarction patients, and calls attention to the possible preventive benefit of beta-blocker therapy against proarrhythmic events experienced in the CAST.


Prognostic Factors for Functional and Anatomic Outcomes in Patients with Diabetic Macular Edema Treated with Dexamethasone Implant.
To investigate the prognostic factors of visual and anatomic outcomes in patients with diabetic macular edema (DME) treated with intravitreal injection of dexamethasone implant. We retrospectively studied 32 eyes of 31 patients with DME for best-corrected visual acuity (BCVA), central macular thickness, and height and width of both intraretinal fluid (IRF) and subretinal fluid. Logistic regression analysis was used to examine correlations between the baseline characteristics and outcomes at 3 and 6 months. Baseline predictor of BCVA ≥20 / 40 at month 3 was short height of baseline IRF (p = 0.02), while good baseline BCVA was a predictor for month 6 (p = 0.01). Predictors of improvement in logarithm of minimum angle of resolution BCVA 0.2 at month 3 were the absence of baseline IRF and poor baseline BCVA (p = 0.02 and p = 0.009, respectively), while poor baseline BCVA was the sole predictor at month 6 (p = 0.01). Predictor of central macular thickness ≤300 μm at month 3 was younger age (p = 0.03), while the absence of IRF was the predictor for BCVA improvement at month 6 (p = 0.02). BCVA ≤20 / 100 at month 3 was predicted by poor baseline BCVA (p = 0.01), and increased width of total IRF was the predictor at month 6 (p = 0.02). Predictor of loss of logarithm of minimum angle of resolution BCVA 0.2 at month 6 was increased width of total IRF at baseline (p = 0.04). Additional injection within 6 months was negatively associated with the presence of baseline DME (p = 0.03). The visual and anatomical outcome of DME treatment with dexamethasone implant can be predicted by baseline visual acuity and IRF morphology.


Pleomorphic Xanthoastrocytoma with Anaplastic Features: Retrospective Case Series.
Pleomorphic xanthoastrocytoma (PXA) is a unique meningocerebral glioma with a relatively favorable prognosis. PXA also possesses a variant with anaplastic features (aPXA), which is associated with poor outcomes. To date, few studies have examined the clinicopathologic importance of these anaplastic features. From 1999-2012, 8 patients with aPXA were treated at the University of California, San Francisco, California, United States. Cases were reconfirmed by neuropathology, and clinical information regarding patient demographics, tumor characteristics, and treatment outcomes was assembled. Tumors were classified as aPXA according to the World Health Organization diagnostic criteria established in 2007. There were 5 female and 3 male patients in our cohort, ranging in age from 4-74 years at initial diagnosis. Seizure was the most common presenting symptom (50%), and the majority of tumors arose in the frontal or temporal lobes (88%). Six patients received subtotal resection (STR), and all suffered from progression despite adjuvant radiotherapy and chemotherapy. Median time to progression was 20 months, with a 1-year progression-free survival rate of 57%. Three aPXA patients expired with a median survival of 87 months. Four patients developed disseminated disease. Three of 8 (38%) showed BRAFv600 mutation. aPXA is associated with poorer clinical outcomes compared with PXA. Gross total resection should be the goal of initial treatment. It remains unclear whether adjuvant radiation and chemotherapy are able to prevent progression or dissemination. Long-term monitoring of all patients is a critical step in management due to the potential for tumors to transform into higher-grade lesions.


Sedentary behaviour is a key determinant of metabolic inflexibility.
Metabolic flexibility is defined as the ability to adapt substrate oxidation rates in response to changes in fuel availability. The inability to switch between the oxidation of lipid and carbohydrate appears to be an important feature of chronic disorders such as obesity and type 2 diabetes. Laboratory assessment of metabolic flexibility has traditionally involved measurement of the respiratory quotient (RQ) by indirect calorimetry during the fasted to fed transition (e.g. mixed meal challenge) or during a hyperinsulinaemic-euglycaemic clamp. Under these controlled experimental conditions, 'metabolic inflexibility' is characterized by lower fasting fat oxidation (higher fasting RQ) and/or an impaired ability to oxidize carbohydrate during feeding or insulin-stimulated conditions (lower postprandial or clamp RQ). This experimental paradigm has provided fundamental information regarding the role of substrate oxidation in the development of obesity and insulin resistance. However, the key determinants of metabolic flexibility among relevant clinical populations remain unclear. Herein, we propose that habitual physical activity levels are a primary determinant of metabolic flexibility. We present evidence demonstrating that high levels of physical activity predict metabolic flexibility, while physical inactivity and sedentary behaviours trigger a state of metabolic 'inflexibility', even among individuals who meet physical activity recommendations. Furthermore, we describe alternative experimental approaches to studying the concept of metabolic flexibility across a range of activity and inactivity. Finally, we address the promising use of strategies that aim to reduce sedentary behaviours as therapy to improve metabolic flexibility and reduce weight gain risk.


Leucobacter zeae sp. nov., isolated from the rhizosphere of maize (Zea mays L.).
A novel yellow-pigmented, aerobic, rod-shaped, non-motile bacterium, designated strain CCMF41T, was isolated from rhizosphere soil of maize (Zea mays) collected in Wufeng District, Taichung, Taiwan. Strain CC-MF41T exhibited 16S rRNA gene sequence similarity of 97.5, 97.3, 97.2 and 97.1% to Leucobacter chironomi MM2LBT (and 'Leucobacter kyeonggiensis'F3-P9 and 'L. humi' Re-6, the names of which have not been validly published), Leucobactertardus K70/01T, L. komagatae IFO 15245T and 'Leucobacter margaritiformis' A23. However,CC-MF41T and 'L. margaritiformis' A23 formed a loosely bound phylogenetic lineage (with alow bootstrap value) associated with species of the genus Leucobacter. In DNA–DNA reassociation experiments, the relatedness of strain CC-MF41T to L. chironomi DSM 19883T was 57.1% (reciprocal value 29.1 %). The DNA G+C content of strain CC-MF41T was 72.1 mol% and the cell-wall peptidoglycan contained 2,4-diaminobutyric acid, alanine, glycine,glutamic acid and threonine. The major menaquinone was MK-11 and the predominant fatty acids were iso-C16 : 0, anteiso-C15 : 0 and anteiso-C17 : 0. The polar lipid profile of strain CCMF41T contained major amounts of diphosphatidylglycerol followed by an unidentified glycolipid, phosphatidylglycerol and an unknown phospholipid. Based on its phylogenetic,phenotypic and chemotaxonomic distinctiveness, strain CC-MF41T represents a novel species of Leucobacter, for which the name Leucobacter zeae sp. nov. is proposed. The type strain isCC-MF41T (=BCRC 80515T=LMG 27265T).


The influence of different preservation and sterilisation steps on the histological properties of amnion allografts--light and scanning electron microscopic studies.
Despite thorough donor screening and preparation under aseptic conditions, conventional methods of preservation do not exclude the probability of a contamination with pathogenic germs. The purpose of this study was to investigate the changes of histological parameters of amnion transplants (ATs) through different methods of sterilisation and preservation. Therefore 10 different procedures for sterilisation and preservation of ATs were described. Specimens of each group were studied using different histological procedures such as light microscopy and scanning electron microscopy. General staining (Haematoxylin-eosin stain, periodic-acid-Schiff, Domack) and immunohistochemical methods have been applied in order to gain additional information concerning the structure of the amniotic epithelium and the basement membrane but also the distribution of collagens and intermediate filaments. Furthermore, the measurement of the ATs thickness was included in order to study the influence of the manufacturing procedures to this property. As a result we found that the histological appearance of the ATs is closely related to the applied sterilisation and preservation procedures. Although the basement membrane remained intact, especially the amniotic epithelium was partially destroyed by irradiation sterilisation. Further, the dissolution of the connective tissue layers into single fibre bundles was clearly visible. Procedures with and without peracetic acid sterilisation (PAA) preserved the tissue structure. Our results showed a significant variation in the tissue's thickness after different preservation procedures. Air- and freeze-dried ATs were found to be the thinnest tissues varying from 20 to 30 microm, the thickest ATs preserved in glycerol varied from 45 to 50 microm. Because ATs showed a preserved tissue structure after PAA sterilisation it can be recommended as an alternative for methods previously described in literature. Depending on the specific use of the AT one may choose from thinner or thicker allografts.


A stretchable conductive Polypyrrole Polydimethylsiloxane device fabricated by simple soft lithography and oxygen plasma treatment.
This paper reports a simple method used to fabricate a stretchable conductive polypyrrole (PPy) rough pore-shape polydimethylsiloxane (p-PDMS) device. An abrasive paper is first used to imprint rough micro-structures on the SU-8 micromold. The p-PDMS microchannel is then fabricated using a standard soft-lithography process. An oxygen plasma treatment is then applied to form an irreversible sealing between the microchannel and a blank cover PDMS. The conductive layer is formed by injecting the PPy mixture into the microchannel which polymerizes in the rough pore-shape micro-structures; The PPy/p-PDMS hybrid device shows good electrical property and stretchability. The electrical properties of different geometrical designs of the PPy/p-PDMS microchannel under stretching were investigated, including straight, curved, and serpentine. Mouse embryonic fibroblasts (NIH/3 T3) were also cultured inside the PPy/p-PDMS device to demonstrate good biocompatibility and feasibility using the conductive and stretchable microchannel in cell culture microfluidics applications. Finally, cyclic stretching and bending tests were performed to evaluate the reliability of PPy/p-PDMS microchannel.


Placing students at the center: the Whole School, Whole Community, Whole Child model.
Students are the heart of the Whole School, Whole Community, Whole Child (WSCC) model. Students are the recipients of programs and services to ensure that they are healthy, safe, engaged, supported, and challenged and also serve as partners in the implementation and dissemination of the WSCC model. A review of the number of students nationwide enjoying the 5 Whole Child tenets reveals severe deficiencies while a review of student-centered approaches, including student engagement and student voice, appears to be one way to remedy these deficiencies. Research in both education and health reveals that giving students a voice and engaging students as partners benefits them by fostering development of skills, improvement in competence, and exertion of control over their lives while simultaneously improving outcomes for their peers and the entire school/organization. Creating meaningful roles for students as allies, decision makers, planners, and consumers shows a commitment to prepare them for the challenges of today and the possibilities of tomorrow.


Latex allergy.
An overview of latex allergy, including methods for preventing reactions and reducing exposure to latex, is provided. Latex allergy is a serious medical problem for an increasing number of patients and health care workers. From an institutional perspective, several precautionary procedures need to be followed to ensure safe care for patients and to protect employees from unnecessary latex exposure. These include establishing a multidisciplinary committee to develop policies and procedures; identifying products containing latex; identifying latex-sensitive patients and employees as well as those at high risk of developing the sensitivity; implementing precautions against allergic reactions to latex; educating personnel and patients on latex allergy; and reviewing quality assurance data to continually improve precautionary procedures. Pharmacists should be involved in developing and implementing these procedures. In particular, they can play a major role in the education of health professionals and patients. There is a strong need for more information on latex sensitivity, especially in the areas of epidemiology, product development, and effectiveness of procedures. Each institution, including its pharmacists, must evaluate the available information about latex sensitivity and determine appropriate actions.


[Isolated laryngeal tuberculosis as an uncommon manifestation of extrapulmonary tuberculosis in adults: a case report].
Isolated laryngeal tuberculosis is rare and sometimes difficult to diagnose. It is the most common cause of laryngeal granuloma. We here report the case of a 58-year-old man, with no particular past medical history, hospitalized due to paroxysmal laryngeal dyspnea, dysphagia to solid foods and dysphonia evolving for 6 months without other associated signs. Laryngoscopic examination showed polyploid formation masking the glottic floor. Histological examination revealed epithelioid and gigantocellular granuloma, without caseous necrosis. Direct microscopic examination and culture were negative. The diagnosis of isolated laryngeal tuberculosis was made based on the endemicity in our country and the absence of other arguments in favor of another type of granulomatosis. Anti-tuberculosis therapy, combined with oral corticosteroids, was indicated based on the presence of severe upper airway edema and symptoms were resolved after 40 days of treatment.


Loss of starch granule initiation has a deleterious effect on the growth of arabidopsis plants due to an accumulation of ADP-glucose.
STARCH SYNTHASE4 (SS4) is required for proper starch granule initiation in Arabidopsis (Arabidopsis thaliana), although SS3 can partially replace its function. Unlike other starch-deficient mutants, ss4 and ss3/ss4 mutants grow poorly even under long-day conditions. They have less chlorophyll and carotenoids than the wild type and lower maximal rates of photosynthesis. There is evidence of photooxidative damage of the photosynthetic apparatus in the mutants from chlorophyll a fluorescence parameters and their high levels of malondialdehyde. Metabolite profiling revealed that ss3/ss4 accumulates over 170 times more ADP-glucose (Glc) than wild-type plants. Restricting ADP-Glc synthesis, by introducing mutations in the plastidial phosphoglucomutase (pgm1) or the small subunit of ADP-Glc pyrophosphorylase (aps1), largely restored photosynthetic capacity and growth in pgm1/ss3/ss4 and aps1/ss3/ss4 triple mutants. It is proposed that the accumulation of ADP-Glc in the ss3/ss4 mutant sequesters a large part of the plastidial pools of adenine nucleotides, which limits photophosphorylation, leading to photooxidative stress, causing the chlorotic and stunted growth phenotypes of the plants.


Comprehensive spatiotemporal mapping of single-cell lineages in developing mouse brain by CRISPR-based barcoding.
A fundamental interest in developmental neuroscience lies in the ability to map the complete single-cell lineages within the brain. To this end, we developed a CRISPR editing-based lineage-specific tracing (CREST) method for clonal tracing in Cre mice. We then used two complementary strategies based on CREST to map single-cell lineages in developing mouse ventral midbrain (vMB). By applying snapshotting CREST (snapCREST), we constructed a spatiotemporal lineage landscape of developing vMB and identified six progenitor archetypes that could represent the principal clonal fates of individual vMB progenitors and three distinct clonal lineages in the floor plate that specified glutamatergic, dopaminergic or both neurons. We further created pandaCREST (progenitor and derivative associating CREST) to associate the transcriptomes of progenitor cells in vivo with their differentiation potentials. We identified multiple origins of dopaminergic neurons and demonstrated that a transcriptome-defined progenitor type comprises heterogeneous progenitors, each with distinct clonal fates and molecular signatures. Therefore, the CREST method and strategies allow comprehensive single-cell lineage analysis that could offer new insights into the molecular programs underlying neural specification.


Arthroscopic treatment of a transarticular low-velocity gunshot wound using tractoscopy.
An unusual case of a close-range, low-velocity gunshot wound to the knee is presented. Arthroscopic debridement was accomplished using standard anterior portals as well as the existing entry and exit wounds. All bullet fragments were successfully removed from the joint by arthroscopy and tractoscopy. A minimally displaced marginal, lateral tibial plateau fracture was observed and treated with restricted weight bearing and active motion. The patient recovered uneventfully and maintained full knee range of motion and a normal gait.


The Role of Virtual Rehabilitation in Total and Unicompartmental Knee Arthroplasty.
This study evaluated the use of telerehabilitation during the postoperative period for patients who underwent total knee arthroplasty (TKA) or unicompartmental knee arthroplasty (UKA). Specifically, this study evaluated the following: (1) patient compliance and adherence to the program, (2) time spent performing physical therapy exercises, (3) the usability of the virtual rehabilitation platform, and (4) clinical outcome scores in a selected primary knee arthroplasty cohort. A total of 157 consecutive patients underwent TKA (n = 18) or UKA (n = 139). These patients used a telerehabilitation system with an instructional avatar, three-dimensional motion measurement and analysis software, and real-time televisit capability designed for arthroplasty patients. Compliance was determined by how many times the patients followed prescribed repetitions of exercises. The total time spent performing exercises for each patient was collected. The usability of the virtual rehabilitation platform (on the patient's end) was evaluated using the system usability scale (SUS) questionnaire. The number of in-person and televisits was recorded for each patient. Patient-reported outcomes were collected through the patient and clinician interfaces and included the Knee Society Score (KSS) for pain and functions, the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score, and Boston University Activity Measure for Post-Acute Care (AM-PAC) score. Patients spent an average of 29.5 days partaking in the therapy. TKA and UKA patients had a mean of 3.5 and 3.2 outpatient follow-up visits, each, for in-office therapy with a physical therapist, respectively. This figure exceeded the mean number of real-time virtual patient-clinician visits by 0.8 visits per patient in the TKA cohort and by 1 visit per patient in the UKA cohort. Patients spent on average 26.5 minutes per day conducting an average of 13.5 exercises. By the end of rehabilitation, patients had spent an average of 10.8 hours performing exercises, and of all the exercises performed, approximately 21 exercises were uniquely designed. Mean SUS score in the cohort was 93 points, which was interpreted as being above the 50th percentile point of the scale. Following therapy, KSS pain and function scores improved markedly and the improvements were measured at 368% for TKA and 350% for UKA (pain) and 27% for UKA and 33% for TKA (function). In addition, WOMAC scores improved by 57% and 66% for UKA and TKA patients while the improvement in AM-PAC scores was at 22% and 24%. This telerehabilitation platform encouraged clinician-patient interaction beyond the hospital setting and offers the advantage of cost savings, convenience, at-home monitoring, and coordination of care, all of which are geared to improve adherence and overall patient satisfaction. Additionally, the biometric data can be used to develop custom physical therapy regimens to assure proper rehabilitation, which is lacking in other telerehabilitation applications that use noninteractive videos that can be watched on mobile devices and tablets.


[Primary obstructive megaureter in infants: medical or surgical treatment? Apropos of 24 cases].
The authors analyse 24 cases of primary obstructive megaureter diagnosed before the age of 3 months. 18 were discovered by antenatal echography between 28 and 39 weeks, whereas 6 were detected by the occurrence of early urinary tract infection. Initial investigations included echography, intravenous urography and micturating cystouretrogram. Primary megaureter was bilateral in 14 cases, unilateral in 12. According to the urographic classification of Beurton, there were 12 ureters stage III, 12 stage II, 10 stage I. 15 patients were operated, 4 of them during the first months (2 reimplantations, 2 urinary diversions). 12 were submitted to later reimplantation at the mean age of 15 months, one had a non functioning kidney removed. 9 were non operated and treated conservatively by antibacterials and regular supervision. Results analysis evaluated according to radiological stage and treatment showed that in more cases, surgery should be differed initially. Complete clinical and radiological resolution is possible even in severe obstructions. Antibacterial treatment beginning at birth favorises regression of dilated ureter. Thus reimplantation should be deferred undercover of regular biological and radiological survey. The age of 12 to 18 months is the most technically favorable time to ureteric reimplantation.


Microsatellites in the endangered species Dyckia distachya (Bromeliaceae) and cross-amplification in other bromeliads.
Microsatellite markers were isolated in Dyckia distachya, an endangered bromeliad from southern Brazil, which will be useful to assess the population genetic structure and reproductive success in introduced and natural populations of this species. Twenty microsatellite loci were developed from an enriched genomic library, and nine of these were amplified. The loci were characterized in 43 individuals from introduced and wild D. distachya populations. All nine loci were polymorphic, with four to ten alleles per locus. In an introduced population the observed and expected heterozygosities ranged from 0.136-0.667 and 0.543-0.877, respectively, while in a wild population it ranged from 0.000 to 0.895 and from 0.050 to 0.811, respectively. The development of these microsatellite markers will contribute to investigations of the reproductive potential and viability of introduced populations of D. distachya as well as the single known wild population. Cross-amplification in other Bromeliaceae species was successful, with high rates in four loci, demonstrating the applicability of these microsatellite markers in other taxa.


Modulation of inwardly rectifying channels by substance P in cholinergic neurones from rat brain in culture.
1. Whole-cell recording was used to investigate the effects of substance P on cultured neurones from the rat nucleus basalis. 2. Brief applications of substance P produced a reduction, about 1 min in duration, of resting membrane conductance. The concentration producing a half-maximal effect was approximately 40 nM, with the continuous presence of substance P resulting in desensitization of the response. 3. The control current-voltage relation exhibited inward rectification over the voltage range -70 to -150 mV, and hyperpolarization produced a time-dependent decrease of current (inactivation). 4. The substance P-sensitive current, obtained by subtracting the current during the presence of the tachykinin from the control current, showed no time-dependent inactivation, though its current-voltage relation also revealed inward rectification, with the reversal potential being approximately equal to the potassium equilibrium potential, Vk. 5. The relation between the substance P-sensitive chord conductance and voltage could be fitted by a Boltzmann equation, with changes in [K+]o shifting this relation along the voltage axis roughly in parallel with the shift in Vk. The maximum conductance was proportional to [( K+]o). 6. Cs+ (0.1 mM) blocked the substance P-sensitive current in a voltage-dependent manner, with an equivalent valency for Cs+ of 1.9. Barium blockage of the substance P-sensitive current was less voltage dependent. 7. Replacement of external Na+ by tetramethylammonium (TMA+) ions reduced the substance P-sensitive current by only 18%. 8. These results indicate that substance P inhibits potassium channels with inward rectifier properties very similar to those of skeletal muscle. 9. Application of sodium nitroprusside did not alter the effect of substance P, suggesting that cyclic GMP plays no role in the channel modulation.


Detection and genomic characterization of motility in Lactobacillus curvatus: confirmation of motility in a species outside the Lactobacillus salivarius clade.
Lactobacillus is the largest genus within the lactic acid bacteria (LAB), with almost 180 species currently identified. Motility has been reported for at least 13 Lactobacillus species, all belonging to the Lactobacillus salivarius clade. Motility in lactobacilli is poorly characterized. It probably confers competitive advantages, such as superior nutrient acquisition and niche colonization, but it could also play an important role in innate immune system activation through flagellin–Toll-like receptor 5 (TLR5) interaction. We now report strong evidence of motility in a species outside the L. salivarius clade, Lactobacillus curvatus (strain NRIC0822). The motility of L. curvatus NRIC 0822 was revealed by phase-contrast microscopy and soft-agar motility assays. Strain NRIC 0822 was motile at temperatures between 15 °C and 37 °C, with a range of different carbohydrates, and under varying atmospheric conditions. We sequenced the L. curvatus NRIC 0822 genome, which revealed that the motility genes are organized in a single operon and that the products are very similar (>98.5% amino acid similarity over >11,000 amino acids) to those encoded by the motility operon of Lactobacillus acidipiscis KCTC 13900 (shown for the first time to be motile also). Moreover, the presence of a large number of mobile genetic elements within and flanking the motility operon of L. curvatus suggests recent horizontal transfer between members of two distinct Lactobacillus clades: L. acidipiscis in the L. salivarius clade and L. curvatus inthe L. sakei clade. This study provides novel phenotypic, genetic, and phylogenetic insights into flagellum-mediated motility in lactobacilli.


A research agenda for public health workforce development.
In the past decades, public health research has focused on categorical rather than cross-cutting or systems issues. Little research has been carried out on the infrastructure required to support public health programs. This article describes the results of an interactive process to develop a research agenda for public health workforce development to inform all those with stakes in the public health system. This research is defined as a multidisciplinary field of inquiry, both basic and applied, that examines the workforce in terms of costs, quality, accessibility, delivery, organization, financing, and outcomes of public health services to increase knowledge and understanding of the relationships among workforce and structure, processes, and effects of public health services. A logic model and five priority research areas resulted from meetings of expert panels during 2000 to 2003. Innovative public and private partnerships will be required to advance cross-cutting and systems-focused research.


Microwave versus cryoablation and radiofrequency ablation for small renal mass: a multicenter comparative analysis.
Ablative techniques emerged as effective alternative to nephron-sparing surgery for treatment of small renal masses. Radiofrequency ablation (RFA) and cryoablation (CRYO) are the two guidelines-recommended techniques. Microwave ablation (MWA) represents a newer technology, less described. The aim of the study was to compare outcomes of MWA to those of CRYO and RFA. Retrospective investigation of patients who underwent MWA, CRYO, or RFA from seven high-volume US and European centers was performed. The first group included patients who underwent CRYO or RFA; the second MWA. We collected baseline characteristics, clinical, intraoperative, and postoperative data. Oncological data included technical success, local recurrence, and progression to metastasis. Multivariate analysis was performed to find predictors for postoperative complications. A composite outcome of "trifecta" was used to assess surgical, functional, and oncological outcomes. 739 patients underwent CRYO or RFA and 50 MWA. CRYO/RFA group had significantly longer operative time (P<0.001), but no difference in LOS, postprocedural Hb mean, intraprocedural complications (P=0.180), overall postprocedural complication rates (P=0.126), and in the 30-day re-admission rate (P=0.853) were detected. No predictive parameter of postprocedural complications was found. Concerning functional outcome, no differences were detected in terms of eGFR at 1 year (P=0.182), ΔeGFR at 1 year (P=0.825) and eGFR at latest follow-up (P=0.070). "Technical success" was achieved in 98.6% of the cases (MWA=100%, CRYO/RFA=98.5%; P=0.775), and there was no significant difference in terms of 2-year recurrence rate (P=0.114) and metastatic progression (P=0.203). Trifecta was achieved in 73.0% of CRYO/RFA vs. 69.6% of MWA cases (P=0.719). MWA is a safe and effective treatment option for small renal masses. Compared with CRYO/RFA, it seems to offer low complication rates, shorter operation time, and equivalent surgical and functional outcomes.


Presence of 2-hydroxymyristate on endotoxins is associated with death in neonates with Enterobacter cloacae complex septic shock.
Enterobacter cloacae complex species are involved in infections among critically ill patients. After a recent E.cloacae outbreak of fulminant neonatal septic shock, we conducted a study to determine whether septic shock severity and its lethal consequence are related to structural features of the endotoxin (lipopolysaccharide [LPS]) of the strains isolated from hospitalized infants and more specifically its lipid A region. It appeared that the LPSs are very heterogeneous, carrying fifteen different molecular species of lipid A. The virulence was correlated with a structural feature identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry and gas chromatography coupled with mass spectrometry: the presence of 2-hydroxymyristic acid as a secondary substituent in lipid A. This is the first published evidence linking LPS structural moiety to neonatal sepsis outcome and opens the possibility of using this fatty acid marker as a detection tool for high-risk patients, which could help reduce their mortality.


Combined treatment of toxic cyanobacteria Microcystis aeruginosa with hydrogen peroxide and microcystin biodegradation agents results in quick toxin elimination.
Under some conditions the growth of toxic cyanobacteria must be controlled by treatment with algicidal compounds. Hydrogen peroxide has been proposed as an efficient and relatively safe chemical which can remove cyanobacteria from the environment selectively, without affecting other microorganisms. However, the uncontrolled release of secondary metabolites, including toxins may occur after such a treatment. Our proposal presented in this paper concerns fast biodegradation of microcystin released after cell lysis induced by hydrogen peroxide. The effectiveness of both, Sphingomonas sp. and heterologously expressed MlrA enzyme, in the removal of the toxin from Microcystis aeruginosa culture was investigated. The results indicate that neither Sphingomonas cells nor MlrA are affected by hydrogen peroxide at the concentrations which stop the growth of cyanobacteria. A several-fold reduction in microcystin levels was documented in the presence of these agents with biodegradation ability. Our results provide evidence that such a combined treatment of water reservoirs dominated by microcystin-producing cyanobacteria may be a promising alternative which allows fast elimination of both, the bloom forming species and toxins, from the environment.


Risk of cancer among firefighters: a quantitative review of selected malignancies.
Using the fixed-effect model, the author quantitatively estimated the risks of cancers of the colon, bladder, kidneys, and brain as well as non-Hodgkin's lymphoma and leukemia among firefighters. The risk of these six cancers was not markedly elevated when cohort mortality studies were considered. When all mortality studies were considered, however, there was a mild increase in risk for kidney cancer and non-Hodgkin's lymphoma, with a summary relative risk (sumRR) of 1.22 (95% confidence interval [CI] = 1.02-1.43) and 1.40 (95% CI = 1.20-1.60), respectively. A subcohort analysis based on duration of employment revealed that firefighters with 30 or more years of employment had a significantly increased mortality risk for colon cancer, sumRR of 1.51 (95% CI = 1.05-2.11); kidney cancer, sumRR of 6.25 (95% CI = 1.70-16.00); brain cancer, sumRR of 2.53 (95% CI = 1.27 7.07); and leukemia, sumRR of 2.87 (95% CI = 1.43-5.14). After firefighters had 40 or more years of employment, their risk of mortality was significantly increased for colon cancer, sumRR of 4.71 (95% CI = 2.03-9.27); kidney cancer, sumRR of 36.12 (95% CI = 4.03-120.42); and bladder cancer, sumRR of 5.7 (95% CI = 1.56-14.63). The risk for non-Hodgkin's lymphoma was elevated but not significantly so among firefighters with 20 or more years of employment, with sumRR of 1.72 (95% CI = 0.90-3.31). Kidney cancer risk was significantly elevated as early as the second decade of employment.


Is and Ought: Descriptive and Prescriptive Cognitions in Military-Related Moral Injury.
Debate exists regarding the most appropriate way to address moral injuries that stem from involvement in war and other military conflicts. In recent years, some researchers have suggested that existing treatments for trauma may be inadequate to address moral injury and have thus proposed novel interventions to help mitigate these concerns. In response, advocates of more traditional approaches have argued that standard trauma interventions are generally sufficient for moral injury, and investment in new interventions may be premature. This conceptual article draws from research findings and current theories of moral injury to demonstrate that there is merit on both sides of this debate, and further clarifying the types of cognitions involved in moral injury can guide effective treatment planning. In particular, the most recent diagnostic criteria for posttraumatic stress disorder is used to distinguish the objectively falsifiable descriptive cognitions often associated with trauma from the subjectively determined prescriptive cognitions that characterize moral injury. Scenarios from war zone deployments have been used to highlight the relevance of this distinction for moral injury, and a general treatment framework that shows how existing and novel interventions may complement one another is presented. Research suggestions for assessing descriptive and prescriptive cognitions in moral injury and empirically validating this treatment framework are discussed.


Mitochondrial Ultrastructure Is Coupled to Synaptic Performance at Axonal Release Sites.
Mitochondrial function in neurons is tightly linked with metabolic and signaling mechanisms that ultimately determine neuronal performance. The subcellular distribution of these organelles is dynamically regulated as they are directed to axonal release sites on demand, but whether mitochondrial internal ultrastructure and molecular properties would reflect the actual performance requirements in a synapse-specific manner, remains to be established. Here, we examined performance-determining ultrastructural features of presynaptic mitochondria in GABAergic and glutamatergic axons of mice and human. Using electron-tomography and super-resolution microscopy we found, that these features were coupled to synaptic strength: mitochondria in boutons with high synaptic activity exhibited an ultrastructure optimized for high rate metabolism and contained higher levels of the respiratory chain protein cytochrome-c (CytC) than mitochondria in boutons with lower activity. The strong, cell type-independent correlation between mitochondrial ultrastructure, molecular fingerprints and synaptic performance suggests that changes in synaptic activity could trigger ultrastructural plasticity of presynaptic mitochondria, likely to adjust their performance to the actual metabolic demand.


Immediate and long-term outcomes of percutaneous transcatheter pulmonary valve implantation.
Transcutaneous pulmonary valve replacement (TPVR) has become an alternative to heart surgery for patients after previous right ventricular outflow tract (RVOT) or pulmonary artery (PA) surgical interventions. The objective was to present immediate and long-term outcomes of trans¬cutaneous pulmonary valve replacement. Between 06/2009 and 06/2016, 46 patients underwent TPVR. Initial diagnoses included tetralogy of Fallot, common arterial trunk, transposition of great arteries post Rastelli correction, left ventricle outflow obstruction after Ross operation, pulmonary atresia, and isolated dysplastic pulmonary valve stenosis. Thirty eight (78%) patients had previously implanted conduits in the pulmonary position, the rest had either RVOT patch reconstruction (n = 6; 13%) or biological valve implantation (n = 2; 4%). They presented primarily with pulmonary stenosis (n = 18; 39%) or regurgitation (n = 28; 60%). All procedures were successful - 44 Melody and 2 Edwards-Sapien valves were implanted. Before each procedure exclusion of potential coronary compression and RVOT prestenting was performed. Significant RVOT systolic gradient reduction (from 35.3 ± 19.5 to 13.5 ± 7.1 mm Hg; p < 0.001) and decrease of right to left ventricle systolic pressure ratio from 0.58 ± 0.18 to mean 0.37 ± 0.1 (p < 0.001) was achieved. Also, in every patient PA-RVOT competence was restored, with minor in¬competence in only a few patients. Post procedure follow-up ranged from 2 to 86 (mean 35.2) months. Follow-up fluoroscopy or chest X-ray revealed 6 stent fractures (2 stent defragmentation - with only 1 significant valve stenosis). Transcutaneous pulmonary valve replacement is a safe procedure with encouraging results, it also enables deferring surgical reintervention in the majority of patients.


Availability of on-site acute vascular interventional radiology techniques performed by trained acute care specialists: A single-emergency center experience.
Comprehensive treatment of a patient in acute medicine and surgery requires the use of both surgical techniques and other treatment methods. Recently, acute vascular interventional radiology techniques (AVIRTs) have become increasingly popular, enabling adequately trained in-house experts to improve the quality of on-site care. After obtaining approval from our institutional ethics committee, we conducted a retrospective study of AVIRT procedures performed by acute care specialists trained in acute medicine and surgery over a 1-year period, including those conducted out of hours. Trained acute care specialists were required to be certified by the Japanese Association of Acute Medicine and to have completed at least 1 year of training as a member of the endovascular team in the radiology department of another university hospital. The study was designed to ensure that at least one of the physicians was available to perform AVIRT within 1 h of a request at any time. Femoral sheath insertion was usually performed by the resident physicians under the guidance of trained acute care specialists. The study sample comprised 77 endovascular procedures for therapeutic AVIRT (trauma, n = 29, and nontrauma, n = 48) among 62 patients (mean age, 64 years; range, 9-88 years), of which 55% were male. Of the procedures, 47% were performed out of hours (trauma, 52%; and nontrauma, 44%). Three patients underwent resuscitative endovascular balloon occlusion of the aorta in the emergency room. No major device-related complications were encountered, and the overall mortality rate within 60 days was 8%. The recorded causes of death included exsanguination (n = 2), pneumonia (n = 2), sepsis (n = 1), and brain death (n = 1). When performed by trained acute care specialists, AVIRT seems to be advantageous for acute on-site care and provides good technical success. Therefore, a standard training program should be established for acute care specialists or trauma surgeons to make these techniques a part of the standard regimen. Therapy/care management study, level V.


Inability to evoke a long-lasting protective immune response to respiratory syncytial virus infection in mice correlates with ineffective nasal antibody responses.
Long-lasting protective antibody is not normally generated in children following primary respiratory syncytial virus (RSV) infection, frequently leading to reinfection. We used the BALB/c mouse model to examine the role of the nasal-associated lymphoid tissue and the bone marrow in the generation of RSV-specific long-lasting plasma cells, with a view to further understanding the mechanisms responsible for the poorly sustained RSV antibody levels following primary infection. We show here that substantial numbers of RSV-specific plasma cells were generated in the bone marrow following challenge, which were maintained thereafter. In contrast, in the nasal-associated lymphoid tissue, RSV-specific plasma cell numbers waned quickly both after primary infection and after challenge and were not maintained at a higher level after boosting. These data indicate that the inability to generate a robust local mucosal response in the nasal tissues may contribute substantially to the likelihood of subsequent reinfection and that the presence of serum anti-RSV antibody without local protection is not enough to protect against reinfection.


Catalytic Formylation of Primary and Secondary Amines with CO2 and H2 Using Abundant-Metal Catalysts.
Catalytic hydrogenation of CO2 is an efficient and selective way to prepare formic acid derivatives, but most of the highly active catalysts used for this purpose require precious metals. In this study, in situ abundant-metal complexes have been evaluated as potential catalysts for CO2 hydrogenation to prepare formamides, including N-formylmorpholine, 2-ethylhexylformamide, and dimethylformamide, from the corresponding amines. From these initial screening results, the most active catalysts for these reactions were found to be MX2/dmpe in situ catalysts (M = Fe(II), Ni(II); X = Cl-, CH3CO2-, acac-; dmpe = 1,2-bis(dimethylphosphino)ethane) in DMSO. The optimal reaction conditions were found to be 100-135 °C and a total pressure of 100 bar. Morpholine was formylated with a TON value of up to 18000, which is the highest TON for the hydrogenation of CO2 to formamides using any abundant-metal-phosphine complex. With an appropriate selection of catalyst and reaction conditions, >90-98% conversion of amine to formamide could be achieved.


Towards the development of a multidisciplinary understanding of the effects of toxic chemical mixtures on health.
Mixtures can be divided into simple (chemicals with comparable properties--health risk assessments on the chemicals) and complex, which can be further subdivided into defined (a reasonably distinct composition, created at a specific time and place despite dissimilar components--risk assessments on the common source) and coincidental (chemicals without similar properties or constant composition in time or space-risk assessments on the receptor). Interactions recognized are: independent action, dose addition (additivity), and potentiation (synergy and antagonism). Unpredicted outcomes need recognition. New approaches in higher education and multidisciplinary investigations are essential. The community of the Society for Environmental Geochemistry and Health should help clarify points such as when transformations in mixtures may become important enough to alter the classification and the risk assessment. The multidisciplinary community is also well placed to support the integration of non-chemical influences into mixture analysis and to contribute to the investigation of cumulative and multiple exposures.


[Age-related specifics or diseases and disability caused by digestive disorders among adults in Saint-Petersburg in 2013-2017.]
The paper presents the retrospective analysis of the structure and dynamics of diseases and disability caused by digestive disorders in Saint Petersburg citizens. The paper presents the results of the comparative analysis of disability among people of active working and pension ages living in Saint Petersburg, versus Russia's average 2013-2018 data. As a result, regularities and differences in the structure and dynamics of digestive disorders in people 18 years old and older, which manifested themselves in after-surgery complications, death rate, and disability rate. The results may be used as reference for making federal or regional-level decisions on developing the system for preventing and early diagnosis of digestive disorders and integrated rehabilitation of disabled persons.


Postoperative Catatonia After Fentanyl, Hydromorphone, and Ketamine Administration in a Patient Taking Sertraline: A Case Report.
Opioid-induced catatonia is underrecognized and poorly understood in the literature. An 81-year-old woman with chronic kidney disease stage III taking sertraline underwent surgery with general anesthesia, receiving fentanyl, hydromorphone, and ketamine. Postoperatively, she was unresponsive, rigid, and cataleptic with pinpoint pupils. Symptoms resolved with a naloxone infusion suggesting opioid-induced catatonia as the leading diagnosis. Differential diagnoses and etiologies discussed reveal a possible multifactorial catatonia mechanism involving opioids, ketamine, and serotonin. Anesthesiologists should consider these potential interactions when using opioids for management of vulnerable patients.


cAMP-dependent ACTH secretagogues facilitate corticotropin releasing activity of angiotensin II on rat anterior pituitary cells in vitro.
Both arginine vasopressin (AVP) and angiotensin II (AII) potentiate the corticotropin-releasing activity of CRF41 via a potentiation of CRF41-induced cAMP production. In perfused rat anterior pituitary cells, AII (10(-8) mol) showed a transitory 2-fold increase in its ACTH-releasing activity, when tested after application extract of rat stalk median eminence. In order to determine whether this facilitating effect on AII corticotropin-releasing activity occurred through cAMP-dependent mechanisms, the ACTH-releasing activity of AII was tested after stimulation with CRF41, AVP or forskolin, three secretagogues with known effects on cAMP production. When given 16 min after CRF41, 10 micrograms/l, AII (10(-8) mol) showed a significant increase (210%) in its ACTH-releasing activity, which returned to the normal level when AII-stimulation was repeated at 32 min (121%) and 48 min (100%). Similarly, forskolin, 3 X 10(-6) mol, produced a significant transitory increase (208%) in the subsequent AII-induced ACTH release whereas AVP, 10 micrograms/l and 100 micrograms/l, had no effect on the following ACTH response to AII. These results suggest that the AII-induced ACTH secretion--which is cAMP independent--nevertheless may be modulated by previously stimulation of the cAMP pathway.


Low-Voltage Zones as the Atrial Fibrillation Substrates: Relationship With Initiation, Perpetuation, and Termination.
Background: Low-voltage zones (LVZs) were usually targeted for ablation in atrial fibrillation (AF). However, its relationship with AF initiation, perpetuation, and termination remains to be studied. This study aimed to explore such relationships. Methods: A total of 126 consecutive AF patients were enrolled, including 71 patients for AF induction protocol and 55 patients for AF termination protocol. Inducible and sustainable AF were defined as induced AF lasting over 30 and 300 s, respectively. Terminable AF was defined as those that could be terminated into sinus rhythm within 1 h after ibutilide administration. Voltage mapping was performed in sinus rhythm for all patients. LVZ was quantified as the percentage of the LVZ area (LVZ%) to the left atrium surface area. Results: The rates of inducible, sustainable, and terminable AF were 29.6, 18.3, and 38.2%, respectively. Inducible AF patients had no significant difference in overall LVZ% compared with uninducible AF patients (10.2 ± 11.8 vs. 8.5 ± 12.6, p = 0.606), while sustainable and interminable AF patients had larger overall LVZ% than unsustainable (16.2 ± 11.5 vs. 0.5 ± 0.7, p < 0.001) and terminable AF patients (44.6 ± 26.4 vs. 26.3 ± 22.3, p < 0.05), respectively. The segmental LVZ distribution pattern was diverse in the different stages of AF. Segmental LVZ% difference was initially observed in the anterior wall for patients with inducible AF, and the septum was further affected in those with sustainable AF, and the roof, posterior wall, and floor were finally affected in those with interminable AF. Conclusions: The associations between LVZ with AF initiation, perpetuation, and termination were different depending on its size and distribution.


Clinical perception of phantom limb sensation in patients with brachial plexus block.
Patients' perception of limb position during regional anaesthesia is frequently incorrect. The incidence and nature of this phenomenon has not yet been completely described. The aim of this prospective study was to assess phantom sensation in patients undergoing a brachial plexus block. Axillary block was performed in 81 patients with the aid of a peripheral nerve stimulator. Immediately after the block, patients were allocated randomly to two groups (Group 1, n = 40; Group 2, n = 41) to have the blocked limb placed either on their thorax or in abduction. Fifteen minutes later, when the block was complete, the position of the limb was changed, without the knowledge of the patient, to a new position of abduction and flexion of the forearm. Patients were questioned about the new limb position. Fifteen minutes later, the limb was then transiently shown to the patient and further placed 'blindly' to another position. Patients were once more questioned about this new position. Correct perception was more frequently observed in both groups following the first interview. Thirty-two and 34 patients gave at least one erroneous response about their limb position when it was initially placed on the thorax or in abduction respectively. Transient visualization of the limb position did not improve the rate of correct response. Patients' perception of the position of the limb after axillary block in the majority of instances was probably due to persisting sensory inputs from the shoulder joint, which is not involved in this block. This is in contrast to supraclavicular or interscalene blocks. A significant number of patients experienced phantom limb sensation. They reported an arm position in fact related to the position of their arm before the axillary block.


Calcitonin gene-related peptide inhibits interleukin-1beta-induced interleukin-8 secretion in human type II alveolar epithelial cells.
Our previous data have shown that type II alveolar epithelial (AEII) cells express neuropeptide calcitonin gene-related peptide (CGRP), and that pro-inflammatory factor interleukin1-beta (IL-1beta) induces CGRP secretion in the A549 human AEII cell line. In the present study, we investigated the effect of endogenous and exogenous CGRP on IL-1beta-induced chemokine interleukin-8 (IL-8) secretion. We used enzyme-linked immunosorbent assay (ELISA) and RT-PCR to detect IL-8 protein and mRNA levels, respectively. siRNA and the stably transfected cell line were used to knock down and overexpress the CGRP gene, respectively, and chemiluminescence assay was used to detect reactive oxygen species (ROS) formation. CGRP-1 receptor antagonist hCGRP8-37 (0.1-1 nmol/L) greatly amplified IL-1beta-induced IL-8 production. The inhibition of CGRP expression by siRNA significantly increased IL-8 secretion upon IL-1beta stimulation. However, cell clones stably transfected with CGRP showed significantly inhibited mRNA and protein levels of IL-8 induced by IL-1beta. These data imply that AEII cell-derived CGRP suppress IL-1beta-induced IL-8 secretion in an autocrine/paracrine mode. Further investigation showed that CGRP attenuated IL-1beta-aroused ROS formation, which is an early indication of pro-inflammatory factor signaling.


Involvement of high-valent manganese-oxo intermediates in oxidation reactions: realisation in nature, nano and molecular systems.
Manganese plays multiple role in many biological redox reactions in which it exists in different oxidation states from Mn(II) to Mn(IV). Among them the high-valent manganese-oxo intermediate plays important role in the activity of certain enzymes and lessons from the natural system provide inspiration for new developments of artificial systems for a sustainable energy supply and various organic conversions. This review describes recent advances and key lessons learned from the nature on high-valent Mn-oxo intermediates. Also we focus on the elemental science developed from the natural system, how the novel strategies are realised in nano particles and molecular sites at heterogeneous and homogeneous reaction conditions respectively. Finally, perspectives on the utilisation of the high-valent manganese-oxo species towards other organic reactions are proposed.


Baseline Susceptibility of Filarial Vector Culex quinquefasciatus (Diptera: Culicidae) to Five Insecticides with Different Modes of Action in Southeast of Iran.
Culex quinquefasciatus (Diptera: Culicidae) is an important vector for many human diseases. The aim of this study was to evaluate the susceptibility level of larval and adult stages of Cu. quinquefasciatus to different groups of WHO recommended insecticides for vector control. Larval stages of the Culex mosquitoes were collected from their natural habitats in Rafsanjan County at Kerman Province, southeast of Iran in 2016. Insecticide susceptibility status of adult female Cx. quinquefasciatus against DDT (4%), deltamethrin (0.05%), malathion 5%, and bendiocarb (0.1%) were determined using WHO standard insecticide susceptibility test. Additional test was carried out to determine the susceptibility status of larvae of Cx. quinquefasciatus to temephos. Bioassay data were analyzed by Probit program. Cx. quinquefasciatus adults showed resistance to all four groups of the tested insecticides according to the WHO criteria for resistance evaluation. The lethal concentrations for 50% mortality (LC50) and 90% mortality (LC90) of temephos against Cx. quinquefasciatus larvae were 0.18mg/l and 0.78mg/l, respectively. This finding also confirms resistance to temephos based on the WHO recommended instructions for resistance evaluation. Resistance to all groups of the tested insecticides should be considered for future vector control investigations in the study area.


Regulation on SIRT1-PGC-1α/Nrf2 pathway together with selective inhibition of aldose reductase makes compound hr5F a potential agent for the treatment of diabetic complications.
(R,E)-N-(3-(2-acetamido-3-(benzyloxy) propanamido)propyl)-2-cyano-3-(4-hydroxy phenyl)acrylamide (hr5F) was design-synthesized based on bioactivity focus strategy as a potential agent to treat diabetic complicates. With in vitro enzyme assay, it is confirmed that hr5F is an effective ALR2 inhibitor with IC50 value of 2.60 ± 0.15 nM, and selectivity index of 86.0 over ALR1, which is a little bit better than the reference Epalrestat (Epa). hr5F inhibits the increase of ALR2 enzyme activity and expression in human lens epithelial cells (HLECs) induced by high glucose. By applying western blot, it was found that hr5F alleviates the high glucose-induced superoxide overproduction insults by regulating SIRT1-PGC-1α/Nrf2 pathway, together with regulating NRF-1, mtTFA, Bax/Bcl-2 to ameliorate cell apoptosis. The in vivo effects of hr5F on short term streptozocin (STZ)-induced diabetic mice confirm the same functions disclosed in vitro. All the evidences support that hr5F may serve as a promising agent in the treatment of diabetic complications with close efficacy and broader indication than the reference Epa.


Imaging-Based Treatment Adaptation in Radiation Oncology.
In many tumor types, significant effort is being put into patient-tailored adaptation of treatment to improve outcome and preferably reduce toxicity. These opportunities first arose with the introduction of modern irradiation techniques (e.g., intensity-modulated radiotherapy) combined with functional imaging for more precise delineation of target volume. On the basis of functional CT, MRI, and PET results, radiation target volumes are altered during the course of treatment, or subvolumes inside the primary tumor are defined to enhance the dosing strategy. Moreover, the probability of complications to normal tissues is predicted using anatomic or functional imaging, such as in the use of CT or PET to predict radiation pneumonitis. Besides focusing, monitoring, and adapting photon therapy for solid tumors, PET also has a role in verifying proton-beam therapy. This article discusses the current state and remaining challenges of imaging-based treatment adaptation in radiation oncology.


Generic Sensor Failure Modeling for Cooperative Systems.
The advent of cooperative systems entails a dynamic composition of their components. As this contrasts current, statically composed systems, new approaches for maintaining their safety are required. In that endeavor, we propose an integration step that evaluates the failure model of shared information in relation to an application's fault tolerance and thereby promises maintainability of such system's safety. However, it also poses new requirements on failure models, which are not fulfilled by state-of-the-art approaches. Consequently, this work presents a mathematically defined generic failure model as well as a processing chain for automatically extracting such failure models from empirical data. By examining data of an Sharp GP2D12 distance sensor, we show that the generic failure model not only fulfills the predefined requirements, but also models failure characteristics appropriately when compared to traditional techniques.