Natural and anthropogenic radionuclides concentration with heavy metals analysis of the sediments collected around Novaya Zemlya.
The Dispersal profile of the radioisotopes (226Ra, 232Th, 235U, 40K, 137Cs) along with potentially toxic elements (Cd, Co, Cr, Cu, Ni, Pb, V, Zn, and Hg) in the sediments around the Novaya Zemlya was determined. The task was fulfilled with the aid of HPGe gamma spectrometry, inductively coupled plasma optical emission spectroscopy, DMA-80 Direct Mercury Analysis System, X-ray diffraction and statistical tools. At most of the locations, the radionuclides activity was higher than the world average activity concentration for the respective nuclei, 40K being the most abundant. From all the potentially toxic elements detected, Cr and Ni were usually observed on higher levels compared to their background values, indicating the probability of the detrimental biological effects. Thus, the present situation at the studied area might be a threat to the neighboring marine life.


Function in vivo of separate segments of the beta subunit of Escherichia coli RNA polymerase.
Transcription of genetic material is catalysed by the enzyme DNA-dependent, RNA polymerase. The multimeric RNA polymerases consist of between 4 and 16 different subunits, of which the two largest, termed beta and beta', are conserved throughout nature. The beta subunit has been implicated in all of the stages of transcription that are catalysed by the complete enzyme. Several lines of evidence have suggested that the function of the beta subunit is not dependent upon the contiguity of the sequence blocks. In this report, a complementary immunological and genetic approach was adopted in order to investigate the individual regions of the beta subunit of RNA polymerase. To this end, the beta structural gene rpoB was separated into four near-equal, non-overlapping segments (as well as 'half' genes) on the basis of 'split' genes in nature, known functional organization and sequence conservation. These segments were used to prepare sequence-specific antibodies against the four individual regions, as well as being expressed in vivo from a tight, lac-controlled high-copy number vector. Immunological probing of the holoenzyme in vitro suggested that the amino-terminal half of the beta polypeptide is buried within the enzyme complex. Of the four segments expressed in vivo, the extreme C-terminal segment was trans-dominant lethal (of the effect of large N-terminal amber fragments on cellular growth; Nene & Glass 1982) and this isolated region was shown to bind the translational elongation factor EF-Tu in vivo. These in vivo and in vitro studies, in conjunction with recent in vitro work (Severinov et al. 1995), unambiguously demonstrate that individual regions of beta may adopt structurally and functionally competent forms, and underline the possibility of in vivo investigation of separate regions of this massive polypeptide chain. A model is presented for the role of EF-Tu in stringent control.


An acridine probe into the physiological state of the cell.
Acridine orange was used as a probe to look into the physiological state of the yeast cell, particularly as regards the change in the properties of the membrane (which acts as a barrier against the incoming acridine orange) and the availability of binding sites for acridine orange in chromosomal DNA during growth. After acridine orange had been introduced into the cell, the genetic change at a specific locus with incubation time was measured photodynamically. A three-fold increase in the rate of penetration of acridine orange into the cell was observed, for instance, in going from the resting phase to the dividing phase. A five-fold increase was observed in the number of binding sites in chromosomal DNA under the same transition of the cell. These two parameters may be useful as a measure of the physiological changes in the cell. Some environmental factors such as pH and temperature were also demonstrated to affect the parameters.


Bone changes in spouses having shared lifestyle for 40 years.
Spousal concordance has already been observed for several other diseases but it has not yet been studied for bone loss. Our aim was to determine whether or not bone changes were the same within spouses who have shared the same environment for many decades. We conducted a prospective study on an average of 5±3 years in a single center. We evaluated in the 104 spouses who came at least twice and had more than 1 year of follow-up the changes in bone mineral density (BMD) at femoral neck adjusted on weight and height, and on hormone replacement therapy use and duration for women. Lifestyle characteristics and biological parameters were evaluated at baseline. The mean (±SD) baseline age of wives was 63±5 years and that of their husbands was 66±5 years. They had been living together for 40±8 years. Most of the environmental baseline factors were correlated within partners: BMI (r=0.26, P<0.01), 25-OHD(3) (r=0.32, P<0.01), daily calories (r=0.52, P<0.001) and calcium intake (r=0.31, P<0.01), physical activity (r=0.43, P<0.0001). Bone loss was observed in wives (-0.0023 g/cm(2)/yr, P<0.01) but not in their husbands (0.0016 g/cm(2)/yr, P=0.10). Changes of BMD were not correlated within spouses (r=0.0004, P=0.99). In women, 25-OHD(3) and interactions of tobacco with 25-OHD(3) and calories intake explained 37% of the bone loss variance (P<0.01). Environmental factors do not appear to influence changes in bone density in the same manner in subjects of different genders.


The misperception of sexual interest.
In the current study (N = 199), we utilized a speed-meeting methodology to investigate misperceptions of sexual interest. This method allowed us to evaluate the magnitude of men's overperception of women's sexual interest, to examine whether and how women misperceive men's sexual interest, and to assess individual differences in susceptibility to sexual misperception. We found strong support for the prediction that women would underestimate men's sexual interest. Men who were more oriented toward short-term mating strategies or who rated themselves more attractive were more likely to overperceive women's sexual interest. The magnitude of men's overperception of women's sexual interest was predicted by the women's physical attractiveness. We discuss implications of gender differences and within-sex individual differences in susceptibility to sexual misperception.


Clinically atypical granular corneal dystrophy with pathologic features of lattice-like amyloid deposits. A study of these families.
Four patients from families in Pennsylvania, Massachusetts, and Argentina were diagnosed clinically as having granular dystrophy. Results of pathologic examination of the corneal buttons from each patient after penetrating keratoplasty confirmed granular deposits in the anterior third of the stroma. Amyloid was demonstrated within some of these granular deposits by Congo red staining with birefringence and dichroism and by electron microscopy. In addition to the morphologically granular deposits, numerous fusiform deposits identified as amyloid by histochemistry and electron microscopy and morphologically identical to those seen in lattice corneal dystrophy were detected deep to the granular deposits. It was further shown that the histochemical pattern of staining of the granular material by a series of lectins was similar to that present in corneas with lattice dystrophy. Although a relationship between these patients cannot be definitively proven, each family traces its origins to the Italian province of Avellino.


The development of direct multicolour fluorescence cross-correlation spectroscopy: towards a new tool for tracking complex biomolecular events in real-time.
Direct three-colour fluorescence cross-correlation spectroscopy can reveal interactions between three fluorescently labelled biomolecules, giving insight toward the complex events that constitute signal transduction pathways. Here we provide the optical and theoretical basis for this technology and demonstrate its ability to detect specific biological associations between nanoparticle-labelled DNA molecules.


Determination of 4-aminobutyric acid, aspartate, glutamate and glutamine and their 13C stable-isotopic enrichment in brain tissue by gas chromatography-mass spectrometry.
A selected-ion monitoring method was developed for measuring 4-aminobutyric acid, aspartate, glutamate, and glutamine in brain tissue. Natural isotopes of these amino acids and their stable-isotopic enrichment following intravenous infusion of a precursor, [13C]glucose, were quantitated. Frozen mouse brain tissue was homogenized in cold 80% ethanol, and the supernatant, equivalent to 1 mg of wet weight brain tissue, was extracted using solid-phase bonded silica ion-exchange columns. Aspartate and glutamate (dicarboxylic acids) were isolated from strong anion-exchange columns, whereas 4-aminobutyric acid and glutamine (neutral amino acids) were isolated from strong-cation exchange columns. n-Butyl ester pentafluoropropionyl amide derivatives of these amino acids were analyzed by gas chromatography-mass spectrometry using a methane positive chemical ionization mode after gas chromatographic separation on a wide-bore, fused-silica capillary column. The method is applicable to determination of brain concentrations of these amino acids as well as their fluxes following administration of a stable-isotopic tracer.


[IXODID TICKS (PARASITIFORMES: IXODIDAE) FROM SMALL MAMMALS IN DEFORESTED BOREAL HABITATS IN THE NORTHERN EUROPEAN RUSSIA].
Changes in the population density of two hard tick species, Ixodes (Exopalpiger) trianguliceps Birula, 1895 and Ixodes persulcatus Schulze, 1930, were examined in 1998-2001, and in 2003-2004 near Gomselga Village (Kondopoga District, 62° 04' N, 33° 55' E) in central Karelia. Data on the abundance of ixodid ticks and the species composition of their hosts in 4 forest sites at different stages of post-felling regeneration (secondary succession), i. e. 7-14, 12-19, 25-32, and 80-87 after logging were obtained. I. persulcatus dominated, comprising 73 % of the total tick number in samples. Regenera- tion of the forest resulted in fluctuations of the population density of two examined tick species: I. (Exopalpiger) trianguliceps (larvae 2.8-5.3; nymphs 1.5-2.2; adults 0-0.09) and I. persulcatus (larvae 4.3-10.6; nymphs 0.6-4.2).


Mechanisms dependent on tryptophan catabolism regulate immune responses in primary Sjögren's syndrome.
To investigate the possible role of tryptophan metabolism in immune regulation of primary Sjögren's syndrome (pSS) the serum concentrations of tryptophan and its metabolite kynurenine were measured by reverse-phase high-performance liquid chromatography (HPLC) in 103 patients with pSS, 56 patients with sicca symptoms and 309 healthy blood donors. The kynurenine per tryptophan ratio (kyn/trp), which reflects the activity of the indoleamine-pyrrole 2,3-dioxygenase (IDO) enzyme involved in tryptophan catabolism, was calculated. Both female and male patients with pSS had significantly higher serum kynurenine concentrations and kyn/trp than subjects with sicca symptoms or healthy blood donors. The median (quartile range) concentration of kynurenine in female patients with pSS was 2.41 micromol/l (1.86-3.26) compared with 1.85 micromol/l (1.58-2.38, P < 0.0001) in subjects with sicca symptoms and 1.96 micromol/l (1.65-2.27, P < 0.0001) in healthy blood donors. Their kyn/trp x 1000 was 34.0 (25.1-44.3) compared with 25.3 (21.1-31.5, P < 0.0001) in subjects with sicca symptoms and 24.3 (21.0-28.9, P < 0.0001) in healthy blood donors. Female pSS patients with high IDO activity (kyn/trp x 1000 > or = 34.0) had significantly higher ESR, serum C-reactive protein, serum IgA and serum beta-2 microglobulin concentrations as well as higher serum creatinine levels, and they had positive antinuclear antibodies more frequently and presented with more American-European consensus group criteria than those with low IDO activity (kyn/trp x 1000 < 34.0). These data suggest that mechanisms dependent on tryptophan catabolism regulate immune responses in pSS. Tryptophan degradation is enhanced in patients with pSS, and high IDO activity is associated with severity of pSS.


The validity of clinical feature profiles for cytomegaloviral anterior segment infection.
Anterior segment cytomegalovirus (CMV) infection, which can be presented as anterior uveitis and corneal endotheliitis, has recently been reported in immunocompetent patients. We would like to access the validity of two presumed characteristic clinical profiles: profile 1, non-herpes simplex virus (HSV)/varicella zoster virus (VZV) corticosteroid-recalcitrant inflammatory ocular hypertensive syndrome (IOHS), and profile 2, corneal endotheliitis with specific coin-shaped keratic precipitates (KPs), that could be helpful in identifying CMV anterior segment intraocular infection. Patients with either profile 1 or profile 2 or both were enrolled consecutively from the uveitis service in Chang Gung Memorial Hospital, Taoyuan, between January 1, 2006 and May 31, 2010. Diagnostic anterior chamber tapping was performed and followed by real-time quantitative polymerase chain reaction (PCR) to detect herpesviridae DNA including HSV I and II, VZV, CMV, and Epstein-Barr virus. Thirty-one eyes of 30 patients (21 males and nine females) were enrolled in this study. CMV DNA PCR was positive in 29 eyes of 28 patients (20 males and eight females). Nineteen of 20 eyes (19 patients) in profile 1 had positive CMV PCR. Ten of 11 eyes (11 patients) in profile 2 had positive CMV PCR. The positive predictive value of profile 1 and profile 2 was 94.7% and 90.9%, respectively. The positive predictive value of combining the two profiles was 93.3%. Non-HSV/ZVZ corticosteroid-recalcitrant IOHS and corneal endotheliitis with specific coin-shaped KPs could be used as the screening tool for CMV anterior segment intraocular infection.


Resource supply and the evolution of public-goods cooperation in bacteria.
Explaining public-goods cooperation is a challenge for evolutionary biology. However, cooperation is expected to more readily evolve if it imposes a smaller cost. Such costs of cooperation are expected to decline with increasing resource supply, an ecological parameter that varies widely in nature. We experimentally tested the effect of resource supply on the evolution of cooperation using two well-studied bacterial public-good traits: biofilm formation by Pseudomonas fluorescens and siderophore production by Pseudomonas aeruginosa. The frequency of cooperative bacteria increased with resource supply in the context of both bacterial public-good traits. In both cases this was due to decreasing costs of investment into public-goods cooperation with increasing resource supply. Our empirical tests with bacteria suggest that public-goods cooperation is likely to increase with increasing resource supply due to reduced costs of cooperation, confirming that resource supply is an important factor in the evolution of cooperation.


Pyrrolopyrrole aza-BODIPY-based NIR-II fluorophores for in vivo dynamic vascular dysfunction visualization of vascular-targeted photodynamic therapy.
Real-time monitoring vascular responses is crucial for evaluating the therapeutic effects of vascular-targeted photodynamic therapy (V-PDT). Herein, we developed a highly-stable and bright aggregation induced emission (AIE) fluorophore (PTPE3 NP) for dynamic fluorescence (FL) imaging of vascular dysfunction beyond 1300 nm window during V-PDT. The superior brightness (ϵmaxΦf>1000 nm ≈ 180.05 M-1 cm-1) and high resolution of PTPE3 NP affords not only high-clarity images of whole-body and local vasculature (hindlimbs, mesentery, and tumor) but also high-speed video imaging for tracking blood circulation process. By virtue of the NPs' prolonged blood circulation time (t1/2 ≈ 86.5 min) and excellent photo/chemical (pH, RONS) stability, mesenteric and tumor vascular dysfunction (thrombosis formation, vessel occlusion, and hemorrhage) can be successfully visualized during V-PDT by FL imaging for the first time. Furthermore, the reduction of blood flow velocity (BFV) can be monitored in real time for precisely evaluating efficacy of V-PDT. These provide a powerful approach for assessing vascular responses during V-PDT and promote the development of advanced fluorophores for biological imaging.


Design, synthesis and biological evaluation of chalcone derivatives as potent and orally active hCYP3A4 inhibitors.
Human cytochrome P450 3A4 (hCYP3A4), one of the most important drug-metabolizing enzymes, catalyze the metabolic clearance of ∼50% therapeutic drugs. CYP3A4 inhibitors have been used for improving the in vivo efficacy of hCYP3A4-substrate drugs. However, most of existing hCYP3A4 inhibitors may trigger serious adverse effects or undesirable effects on endogenous metabolism. This study aimed to discover potent and orally active hCYP3A4 inhibitors from chalcone derivatives and to test their anti-hCYP3A4 effects both in vitro and in vivo. Following three rounds of screening and structural optimization, the isoquinoline chalcones were found with excellently anti-hCYP3A4 effects. SAR studies showed that introducing an isoquinoline ring on the A-ring significantly enhanced anti-CYP3A4 effect, generating A10 (IC50 = 102.10 nM) as a promising lead compound. The 2nd round of SAR studies showed that introducing a substituent group at the para position of the carbonyl group on B-ring strongly improved the anti-CYP3A4 effect. As a result, C6 was identified as the most potent hCYP3A4 inhibitor (IC50 = 43.93 nM) in human liver microsomes (HLMs). C6 also displayed potent anti-hCYP3A4 effect in living cells (IC50 = 153.00 nM), which was superior to the positive inhibitor ketoconazole (IC50 = 251.00 nM). Mechanistic studies revealed that C6 could potently inhibit CYP3A4-catalyzed N-ethyl-1,8-naphthalimide (NEN) hydroxylation in a competitive manner (Ki = 30.00 nM). Moreover, C6 exhibited suitable metabolic stability in HLMs and showed good safety profiles in mice. In vivo tests demonstrated that C6 (100 mg/kg, orally administration) significantly increased the AUC(0-inf) of midazolam by 3.63-fold, and strongly prolonged its half-life by 1.66-fold compared with the vehicle group in mice. Collectively, our findings revealed the SARs of chalcone derivatives as hCYP3A4 inhibitors and offered several potent chalcone-type hCYP3A4 inhibitors, while C6 could serve as a good lead compound for developing novel, orally active CYP3A4 inhibitors with improved druglikeness properties.


Prediction of graft flow impairment by intraoperative transit time flow measurement in off-pump coronary artery bypass using arterial grafts.
We assessed the validity of intraoperative transit time flow measurement (TTFM) in predicting graft flow abnormalities. Intraoperative graft flow measurement using TTFM and early postoperative coronary angiography was performed in 58 patients who underwent total arterial off-pump coronary artery bypass. Five variables (flow pattern, mean flow, pulsatility index, insufficiency ratio, and fast Fourier transformation ratio) were measured and compared between 103 normal and 14 abnormal (occluded or competitive) grafts. The grafts anastomosed to the right coronary territories showed significantly less diastolic dominant pattern, lower mean flow and fast Fourier transformation ratio, and higher pulsatility index than grafts to the left coronary artery territories (p < 0.05). None of the abnormal grafts showed a diastolic dominant flow pattern. The abnormal grafts demonstrated significantly lower mean flow and fast Fourier transformation ratio and higher pulsatility index and insufficiency ratio than normal grafts (p < 0.05). When our criteria for detection of abnormal graft flow, [(1) systolic dominant or balanced pattern of the flow curve in the left coronary territories, systolic dominant pattern of the flow curve in the right coronary territories; (2) mean flow < 15 mL/min; (3) pulsatility index > 3 in the left coronary territories and > 5 in the right coronary territories; and (4) insufficiency ratio > 2%] were applied, the sensitivity and specificity of TTFM to detect the graft flow abnormality were 96.2% and 76.9%, respectively. Our data suggest that TTFM is a reliable intraoperative tool to predict graft flow impairment.


Plasma L-arginine levels in a rabbit model of hypercholesterolaemia.
A novel method for the quantitation of L-arginine in plasma samples has been developed, based on the technique of enantiomer labelling coupled to the capillary gas chromatographic separation of amino acids. Using this technique plasma L-arginine levels have been measured in rabbits fed a 1% cholesterol-containing diet and compared with rabbits fed a standard diet. Blood cholesterol levels were significantly elevated in the rabbits fed cholesterol but no significant difference was found in plasma L-arginine levels between the cholesterol-fed and control animals. No D-arginine was found in any of the plasma samples. It is concluded that the impairment in endothelium-dependent relaxation previously described in this animal model of hypercholesterolaemia is not due to deficiency of plasma L-arginine.


CSF rhinorrhea: detection and localization using overpressure cisternography with Tc-99m-DTPA.
We performed 32 overpressure radionuclide cisternography (ORNC) studies to examine 26 patients who were clinically suspected of having cerebrospinal fluid (CSF) fistula with rhinorrhea. Fifteen (47%) of these cisternography studies were positive, and the site of the leak was identified. No leak could be demonstrated in the other 17. Of 23 examinations performed in patients who had clinically documented CSF rhinorrhea, 15 (65%) were scintigraphically positive. The rapid cephalad transit of the radionuclide bolus allowed completion of the study within 30 to 45 minutes. Seven examinations were also performed with overpressure metrizamide CT cisternography (OMCTC), and five demonstrated concordant results with the radionuclide study. Patient discomfort and side effects were minimal. We conclude that radionuclide infusion cisternography is a safe, rapid, and accurate method of investigating a suspected or proven CSF rhinorrhea and that it is complementary to metrizamide cisternography.


Silver Nanoparticles-Cow Dung Combination Disrupts Physiology, Enzyme Activities with Corresponding Increased Oxidative Stress and Heavy Metal Accumulation in Abelmoschus esculentus.
The effects of a silver nanoparticles (AgNPs)-cow-dung combination on Abelmoschus esculentus physiology, enzyme activities, heavy metal bioaccumulation, and micronutrients were investigated. A. esculentus seedlings were nurtured with foliar application of water, 0.75 mg/L each of AgNPs, cow-dung and AgNPs-cow dung till maturity. AgNPs-cow dung inhibited root and shoot lengths by 12.9% and 24.8%, respectively as opposed to elongation recorded for individual application compared to the control. In contrast to the promotion of photosynthesis, enzyme and antioxidant activities by individual applications, AgNPs-cow dung suppressed more than 50% of these parameters. Implied toxicities of AgNPs-cow dung on A. esculentus manifested in increased malondialdehyde contents with concomitant higher bioaccumulation factor of heavy metals. Cow-dung and AgNPs each stimulated A. esculentus activities in different ways, but their combination was inhibitory, which may be ascribed to the transformed AgNPs in cow-dung and relative availability of toxic metals.


MicroRNA hsa-miR-4674 in Hemolysis-Free Blood Plasma Is Associated with Distant Metastases of Prostatic Cancer.
We analyzed microRNA profile in hemolysis-free blood plasma of patients with prostatic cancer. The metastatic form of prostatic cancer was found to be associated with increased levels of hsa-miR-22-3p, hsa-miR-663a, and hsa-miR-4674 in comparison with non-metastatic form. Common candidate target genes of these microRNA include JUNB, KMT2A, and XPO6.


In silico 3D structure prediction and hydrogen peroxide binding study of wheat catalase.
Catalase (CAT) is one of the most active enzyme catalysts found in plants, animals and in all aerobic microorganisms. The major function of the enzyme is to decompose H2O2, produced by cellular metabolic activities under normal and stressful conditions to water and oxygen. The present study involves 3D structure modeling of wheat catalase from Triticum aestivum by MODELLER9v7 and its binding study with H2O2. The Evaluation of the model was based on Discrete Optimized Protein Energy Score (DOPE). The structure was also validated using PROCHECK comprising of 95.0% amino acid residues in favored regions of Ramachandran plot, Verify3D and ProsA which confirm that the model is reliable. The 3D model of the molecule was found to consist of ten strands and seventeen helices having a common fold characterised by β-pleated sheet flanked either side by helices. The docking study with H2O2 indicates that Gln352 and Arg353 are two important determinant residues in binding H2O2 as these residues have strong hydrogen bonding contacts with the substrate. These hydrogen-bonding interactions play a significant role in the stability of the complex.


The role of human factors in the design and development of an insulin pump.
This article discusses human factors (HF) processes and how they are applied during the development of a medical device to minimize the risk that the user interface design could lead to patient errors, adverse events, and product recalls. This process is best defined as "prevention through design." The HF design process is exemplified by three distinct phases: (1) preliminary analysis, (2) formative design evaluation and modification, and (3) design validation. Additional benefits of employing HF principles during medical device development are briefly reviewed, including reduced patient risk by eliminating design flaws, increased patient adherence through the reduction in the complexity of therapeutic regimes, and reduced likelihood for product recalls.


The neural encoding of cocaine-induced devaluation in the ventral pallidum.
Cocaine experience affects motivation structures such as the nucleus accumbens (NAc) and its major output target, the ventral pallidum (VP). Previous studies demonstrated that both NAc activity and hedonic responses change reliably as a taste cue comes to predict cocaine availability. Here we extended this investigation to examine drug-experience induced changes in hedonic encoding in the VP. VP activity was first characterized in adult male Sprague-Dawley rats in response to intraoral infusions of palatable saccharin and unpalatable quinine solutions. Next, rats received 7 daily pairings of saccharin that predicted either a cocaine (20mg/kg, ip) or saline injection. Finally, the responses to saccharin and quinine were again assessed. Of 109 units recorded in 11 rats that received saccharin-cocaine pairings, 71% of responsive units significantly reduced firing rate during saccharin infusions and 64% increased firing rate during quinine exposure. However, as saccharin came to predict cocaine, and elicited aversive taste reactivity, VP responses changed to resemble quinine. After conditioning, 70% of saccharin-responsive units increased firing rate. Most units that encoded the palatable taste (predominantly reduced firing rate) were located in the anterior VP, while most units that were responsive to aversive tastes were located in the posterior VP. This study reveals an anatomical complexity to the nature of hedonic encoding in the VP.


Type 1 diabetes islet associated antibodies in subjects infected by echovirus 16.
To determine whether the emergent infection by echovirus 16 that occurred in Cuba during the year 2000 was related to the presence of Type 1 diabetes associated autoantibodies. The presence of ICA, IAA, GADA, IA2 antibodies and neutralizing antibodies (NtAb) to echovirus 16 were determined in sera from 38 infected children and adolescents and 80 control subjects, matched in sex, age, local residence and time of sample collection. The occurrence of a large-scale echovirus 16 epidemic was associated with the appearance of humoral autoimmune markers of Type 1 diabetes, especially for ICA, IAA and GADA. In the convalescent stage, ICA, IAA and GADA seroconversion was shown in 92.1%, 44.7% and 28.9% of echovirus 16 infected subjects. None of the 80 uninfected subjects had ICA or IAA, while one was GADA positive. ICA, IAA and GADA frequency was higher in the convalescent than in the acute stage (p<0.0005). A strong positive correlation was found between the NtAb to echovirus 16 and ICA titres in both acute and convalescent stage (r=0.91; p<0.0001, r=0.55; p=0.0003 respectively). This work provides evidence of an association between echovirus 16 infection and the presence of Type 1 diabetes related antibodies (ICA, IAA and GADA). Our data show that the echovirus 16 infection might be capable of inducing a process of autoimmune beta-cell damage and support the hypothesis that enterovirus infections are important risk factors for the development of Type 1 diabetes.


Baicalein, isolated from Scutellaria baicalensis, protects against endothelin-1-induced pulmonary artery smooth muscle cell proliferation via inhibition of TRPC1 channel expression.
We investigated the antiproliferative effects of baicalein, isolated from Scutellaria baicalensis (Huang-qin), on ET-1-mediated pulmonary artery smooth muscle cells (PASMCs) proliferation and the mechanisms underlying these effects. Intrapulmonary artery smooth muscle cells were isolated and cultured from female Sprague-Dawley rats and used during passages 3-6. The proliferation of PASMCs was quantified by cell counting and XTT assay. The protein expression of TRPC1 and PKCα were determined by western blotting. The cell cycle pattern was assayed by flow cytometry. The intracellular calcium concentrations ([Ca(2+)](i)) were measured using the fluorescent indicator fura-2-AM and flow cytometry. Baicalein (0.3-3 μM) inhibited PASMCs proliferation, promoted cell cycle progression, enhanced [Ca(2+)](i) levels, increased capacitative Ca(2+) entry (CCE), upregulated the canonical transient receptor potential 1 (TRPC1) channel and membrane protein kinase Cα (PKCα) expression induced by ET-1 (0.1 μM). The PKC activator PMA (1 μM) reversed the inhibitory effects of baicalein on ET-1-induced upregulation of TRPC1 expression and S phase accumulation, while the PKC inhibitor chelerythrine (1 μM) potentiated baicalein-mediated G(2)/M phase arrest and TRPC1 channel inhibition. Our findings suggest that baicalein protects against ET-1-induced PASMCs proliferation via modulation of the PKC-mediated TRPC channel.


[Role of computed tomography in the preoperative diagnosis of otosclerosis].
Otosclerosis is an osteodystrophy of the otic capsule and presents with progressive conductive hearing loss. Imaging studies, especially computed tomography (CT) and cone-beam CT, have gained increased relevance in the diagnosis of otosclerosis. This study investigated whether there is a correlation between the extent of otosclerosis in high-resolution or cone-beam CT and hearing loss in pure-tone audiometry. Based on an existing classification of otosclerotic foci, a classification was established. Preoperative CT scans of patients undergoing stapedotomy between 2015 and 2019 were evaluated and classified by two independent otorhinolaryngologists. The preoperative pure-tone audiograms were analysed and compared to the results of CT. A total of 168 CT studies (i.e., 168 ears) in 156 patients with intraoperatively confirmed otosclerosis were included in our study. A correlation between the extent of the otosclerotic focus or the calculated scores and hearing loss in pure-tone audiometry (air conduction, bone conduction and air-bone-gap) could not be proven. Preoperative CT is not obligatory. However, preoperative imaging using CT or cone-beam CT can be helpful to confirm the diagnosis and exclude other middle or inner ear pathologies as well as in planning of the surgical procedure in the overall context of otoscopy and audiometry. A correlation with the degree of hearing impairment could not be demonstrated and remains unclear.


Corrigenda: Table Correction. Nonalcoholic Fatty Liver Disease in Diabetes. Part I: Epidemiology and Diagnosis.
This corrects the article on p. 31 in vol. 43, PMID: 30793550.


In vitro models for assessing therapeutic angiogenesis.
Arterial obstruction leading to ischemia causes a reduction of oxygen and nutrient supply to distal tissues. The physiological response to tissue ischemia triggers a cascade of events that results in the development of accessory vasculature to increase local tissue perfusion and to salvage tissue. However, this adaptive mechanism of repair is suboptimal in some patients. Therapeutic angiogenesis aims to stimulate new blood vessel formation via the local administration of proangiogenic agents or cell therapy products (CTPs). In this review, we provide a summary of the current understanding of in vitro models for assessing the angiogenic potential of a product.


Molecular cloning and characterization of the alkB gene of Escherichia coli.
Using methods of in vitro recombination we constructed hybrid plasmids that can suppress the increased methylmethane sulfonate sensitivity caused by alkB mutation. Since the cloned DNA fragment was mapped at 47 min on the Escherichia coli K12 genetic map, an area where the alkB gene is located, we concluded that the cloned DNA fragment contains the alkB gene itself but not other genes that suppress alkB mutation. Specific labeling of plasmid-encoded proteins by the maxicell method revealed that the alkB codes for a polypeptide with a molecular weight of about 27,000. Introduction of a small deletion into the alkB region of the bacterial chromosome resulted in inactivation of both the alkB and ada genes, thereby suggesting that the two genes are adjacent on the E. coli chromosome.


Approach for comparing protein structures and origami models.
The research fields of proteins and origami have intersected in the study of folding and de-novo design of proteins. However, there is limited knowledge on the analogy between protein structures and origami models. We propose a general approach for comparing protein structures with origami models, and present a test case, comparing transmembrane β-barrel and α-helical barrel with the Yoshimura and Kresling origami models. While both shapes and structures may look similar, we demonstrated that the β-barrel and the α-helical barrel are in agreement only with the shape and structural characteristics of the Kresling model. Through the analogy, it is explained how the structural characteristic can help the β-barrel and α-helical barrel to adjust length and diameter in response to changes in the membrane structure. However, such conformations only apply to the α-helical barrel, and the β-barrel, in spite of resembles to the Kresling model, remains stiff due to hydrogen bonds between the β-strands. Thus, our analysis suggests that there are similar patterns between protein structures and origami models and that the proposed approach may provide important insight on the role that the structure of a protein fulfils, and on the preferred structural design of novel proteins with unique characteristics.


A new method for recombinant adeno-associated virus vector delivery to murine diaphragm.
Genetically modified mice are important models for evaluation of potential gene therapies for human diseases. However, their small size often precludes the use of clinically feasible methods for vector delivery, therefore, alternative methods must be used. We have developed a gel-based method for delivery of recombinant adeno-associated virus vectors to the mouse diaphragm, an important target organ for many myopathic diseases. We hypothesized that delivery of vectors in a viscous solution would increase transduction by providing a longer exposure time to target cells. We demonstrate that gel-mediated delivery of rAAV serotypes 1, 2, and 5 results in higher transduction efficiencies than free vectors alone when administered in vivo to mouse diaphragms. We further establish greater tropism of rAAV1 vectors for the diaphragm compared to serotypes 2 and 5. This report describes a novel method for efficient delivery of rAAV vectors to the mouse diaphragm and is the first demonstration of gene transfer to the diaphragm using recombinant adeno-associated virus vectors.


Growth and metabolic characteristics of fastidious meat-derived Lactobacillus algidus strains.
Lactobacillus algidus is a meat spoilage bacterium often dominating the bacterial communities on chilled, packaged meat. Yet, L. algidus strains are rarely recovered from meat, and only few studies have focused on this species. The main reason limiting detailed studies on L. algidus is related to its poor growth on the media routinely used for culturing food spoilage bacteria. Thus, our study sought to develop reliable culture media for L. algidus to enable its recovery from meat, and to allow subculturing and phenotypic analyses of the strains. We assessed the growth of meat-derived L. algidus strains on common culture media and their modifications, and explored the suitability of potential media for the recovery of L. algidus from meat. Moreover, we determined whether 12 meat-derived L. algidus strains selected from our culture collection produce biogenic amines that may compromise safety or quality of meat, and finally, sequenced de novo and annotated the genomes of two meat-derived L. algidus strains to uncover genes and metabolic pathways relevant for phenotypic traits observed. MRS agar supplemented with complex substances (peptone, meat and yeast extract, liver digest) supported the growth of L. algidus, and allowed the recovery of new L. algidus isolates from meat. However, most strains grew poorly on standard MRS agar and on general-purpose media. In MRS broth, most strains grew well but a subset of strains required supplementation of MRS broth with additional cysteine. Supplementation of MRS broth with catalase allowed growth in aerated cultures suggesting that the strains produced hydrogen peroxide when grown aerobically. The strains tested (n = 12) produced ornithine from arginine and putrescine from agmatine, and two strains produced tyramine from tyrosine. Our findings reveal that L. algidus populations are underestimated if routine culture protocols are applied, and prompt concerns that L. algidus may generate tyramine or putrescine in meat or fermented meat products.


Increased frequency of N-methyl-N'-nitro-N-nitrosoguanidine-induced nuclear anomalies in the forestomach of rats pretreated with sodium chloride.
The frequency of nuclear anomalies (micronuclei, pyknosis, and karyorrhexis) in the forestomach mucosa was examined in Sprague-Dawley male rats given a single oral dose of 50 or 150 mg/kg of the gastric carcinogen N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) 17 h after the administration of 2 ml of a 3 M NaCl solution. Rats pretreated with NaCl displayed an incidence of nuclear anomalies approximately 3-fold greater than the one observed in rats given MNNG alone, and micronucleated cells accounted to a significant extent for this increase. These findings confirm that NaCl presumably acts as co-carcinogen in the initial phase of gastric carcinogenesis, and suggest that its administration before the carcinogen might increase the sensitivity of short-term tests for the preliminary screening of potential gastric carcinogens.


Abdominal radical trachelectomy for treatment of deep infiltrating endometriosis of the cervix.
Surgical management of deep infiltrating endometriosis can be very challenging even for experienced gynecologists. Radical procedures like bowel resection and nephrectomy have been reported for treatment of the disease. Our aim is to report abdominal radical trachelectomy for treatment of deep infiltrating endometriosis of the cervix causing obstructive uropathy and diminished kidney function. We present a 38-year-old woman who was treated in our department for cervical endometriosis involving the vagina and left parametrium. Abdominal radical trachelectomy, insertion of a pig-tail catheter in the left ureter and end-to-end anastomosis of the uterus with the vagina was performed to remove the endometriotic lesion. Cooperation between gynecologists, urologists and nephrologists enabled fertility preservation as well as improvement of renal function. Deep infiltrating endometriosis is a complex disease that requires a multidisciplinary approach. Abdominal radical trachelectomy for cervical lesions seems feasible in this setting and helps preserve fertility.


Thimerosal hyperpolarizes arterial smooth muscles in an endothelium-dependent manner.
Thimerosal activates the production of endothelium-derived relaxing factor (EDRF). I examined whether thimerosal also causes the release of an endothelium-dependent hyperpolarizing factor (EDHF). Thimerosal caused an endothelium-dependent hyperpolarization of smooth muscle. This effect is unlikely to be caused by the property of thimerosal to inhibit the acyl-coenzyme A: lysolecithin acyltransferase (LAT) since neither arachidonic acid nor lysolecithin hyperpolarized the muscles. I conclude that thimerosal is not a pure activator of EDRF (nitric oxide) production, but that it also releases a distinct EDHF.


Predictors of functional outcome after spinal cord surgery: Relevance of intraoperative neurophysiological monitoring combined with preoperative neurophysiological and MRI assessments.
To assess the accuracy of intraoperative neurophysiological monitoring (IONM) in predicting immediate and 3-month postoperative neurological new deficit (or deterioration) in patients benefiting from spinal cord (SC) surgery; and to identify factors associated with a higher risk of postoperative clinical worsening. Consecutive patients who underwent SC surgery with IONM were included. Pre and postoperative clinical (modified McCormick scale), radiological (lesion-occupying area ratio), and electrophysiological features were collected. A total of 99 patients were included: 14 (14.1%) underwent extradural surgery, 50 (50.5%) intradural extramedullary surgery, and 35 (35.4%) intramedullary surgery. Cumulatively, multimodal IONM (motor and somatosensory evoked potentials, D-wave whenever possible) significantly predicted postoperative deficits (p<0.001), with a sensitivity, specificity, positive predictive value, and negative predictive value of 0.81, 0.93, 0.83, and 0.92, respectively. Sixty (60.6%) patients displayed no IONM change, whereas 39 (39.4%) displayed IONM worsening. In multivariate analysis, predictors for postoperative clinical worsening were: abnormal preoperative electrophysiological assessment (p=0.03), intramedullary tumor (p<0.001), lesion-occupying area ratio ≥0.7 (p<0.001), and IONM alterations (p<0.001). Three months after the surgical procedure, in patients presenting at least one of the risk factors described above, 45/81 (55.6%) and 19/81 (23.5%) were clinically and electrophysiologically improved, respectively; while 13/81 (16.0%) and 10/81 (12.3%) were clinically and electrophysiologically worsened. Multimodal IONM is an essential tool to guide SC surgery, and enables the accurate prediction of postoperative neurological outcome. Specific attention should be given to patients presenting with preoperative electrophysiological abnormalities, large tumor volume, and intramedullary tumor location.


New catalyst design for polymerization of norbornene esters by reducing intramolecular interaction.
We have used density functional theory to study palladium-based catalysts commonly used for the polymerization of norbornene derivatives with an ester group. Exo-exo, exo-endo, and endo-endo isomers of catalyst complexes were investigated; the endo-endo isomer was the most stable and inactive due to an intramolecular interaction between Pd and O of the carbonyl group. Phosphine groups are effective in minimizing the Pd-O interaction in the endo-endo isomer and P(C6H11)3 was found to be the most efficient reagent. The intramolecular Pd-O interactions were estimated using model complexes, and it was demonstrated that they play a crucial role in stabilizing the endo-endo isomer.


Emergency intravenous pyelography in the setting of possible renal colic: is it indicated?
To determine whether emergency IV pyelography (IVP) adds significant information to clinical judgment in the setting of possible renal colic. A prospective voluntary survey of a convenience sample of physicians at the time of patient encounter. The emergency department of a university hospital, annual census 50,000 visits. Emergency medicine faculty physicians. Physicians were surveyed before and after IVP was performed on patients with possible renal colic. Over the course of 12 months, 62 patients with possible kidney stones were evaluated by 14 different faculty ED physicians, who filled out surveys. Before IVP results were obtained, 63% of patients were thought to have a high (more than 75%) probability of ureteral obstruction. In 59% of these patients, the IVP results showed ureteral obstruction. The IVP revealed unexpected findings in 42% of all patients, including normal results in 19%, ureteral stones in 5%, higher than expected grade of obstruction in 6%, and lower than expected grade of obstruction in 6%, and lower than expected grade of obstruction in 5%. Management was reported to have been changed in 60% of all patients for a wide variety of reasons, but consideration of alternative diagnosis was the most common reason, occurring in 23%. Management changes included unexpected hospital admission in five patients (9%) and emergency urologic consultation in five patients (9%). Emergency IVPs are useful in the evaluation of patients with suspected renal colic, primarily when they rule out ureteral obstruction and allow consideration of alternative diagnoses. Not uncommonly, emergency urologic consultation or hospitalization occurs on the basis of IVP findings.


Cheleionwatanabei sp. n., a new species of Stereomerini (Coleoptera, Scarabaeidae, Aphodiinae), and description of the male of C.jendeki.
Cheleionwatanabei sp. n. is described from Pahang, Peninsular Malaysia and represents the third species of the genus Cheleion Vårdal & Forshage, 2010 (Coleoptera, Scarabaeidae, Aphodiinae, Stereomerini). A description of the previously unknown male of C.jendeki Král & Hájek, 2015, and a key to the species of the genus Cheleion are also provided.


Ultrasonic monitoring of erosion/corrosion thinning rates in industrial piping systems.
Monitoring pipe wall erosion/corrosion thinning rates is an important issue in petrochemical and power generation industries. In this paper, two signal processing techniques are utilized for estimating the thinning rate based on ultrasonic pipe wall thickness data collected over a short period of time. The first is a combination of cross-correlation and polynomial curve fitting and the second is a model-based estimation (MBE) scheme. These techniques are applied to data collected from an accelerated thinning rate apparatus and both show that they are capable of estimating the thinning rates quickly in short time periods with good accuracy. In laboratory applications, thinning rates as low as 10 μm/year were measured within 15 days with an uncertainty of ±1.5 μm/year by both techniques. Although the MBE technique can yield marginally better accuracy, the greater stability and computational speed of the cross-correlation technique make it the preferred choice for industrial use.


Small Molecules Enhance Reprogramming of Adult Human Dermal Fibroblasts to Dorsal Forebrain Precursor Cells.
The development of human cell-based platforms for disease modeling, drug discovery, and regenerative therapy relies on robust and practical methods to derive high yields of relevant neuronal subtypes. Direct reprogramming strategies have sought to provide a means of deriving human neurons that mitigate the low conversion efficiencies, and protracted timing of human embryonic stem cell and induced pluripotent stem cell-derived neuron specification in vitro. However, few studies have demonstrated the direct conversion of adult human fibroblasts into multipotent neural precursors with the capacity to differentiate into cortical neurons with high efficiency. In this study, we demonstrate a reprogramming strategy using chemically modified mRNA encoding the proneural genes SOX2 and PAX6 coupled with small molecule supplementation to enhance the derivation of human-induced dorsal forebrain precursors directly from adult human dermal fibroblasts (aHDFs). Through transcriptional and phenotypic analysis of lineage-specific precursor and cortical neuron markers, we have demonstrated that this combined strategy significantly enhances the direct derivation of dorsal forebrain precursors from aHDFs, which, after timely exposure to defined differentiation media, gives rise to high yields of functional glutamatergic neurons. We propose that this combined strategy provides a highly tractable and efficient human cell-based platform for disease modeling and drug discovery.


Isolation, purification, and identification of antialgal substances in green alga Ulva prolifera for antialgal activity against the common harmful red tide microalgae.
Ten compounds (1~10) were successfully isolated from green algae Ulva prolifera through the combination of silica gel column chromatography, Sephadex LH-20 column chromatography and repeated preparative thin-layer chromatography. These ten compounds showed antialgal activity against red tide microalgae. Among them, compounds 3, 6, and 7 showed stronger antialgal activity against red tide microalgae. Furthermore, their structure was identified on the basis of spectroscopic data. There are three glycoglycerolipids: 1-O-octadecanoic acid-3-O-β-D-galactopyranosyl glycerol (2), 1-O-palmitoyl-3-O-β-D-galactopyranosyl glycerol (4), and 1-O-palmitoyl-2-O-oleoyl-3-O-β-D-galactopyranosyl glycerol (5); two monoglycerides: glycerol monopalmitate (1), 9-hexadecenoic acid, 2,3-dihydroxypropyl ester (3); two terpenoids: loliolide (6) and lsololiolide (7); one lipid-soluble pigments: zeaxanthin (8); one sterol: cholest-5-en-3-ol (9); and one alkaloid: pyrrolopiperazine-2,5-dione (10). These compounds were isolated from U. prolifera for the first time, and compounds 2, 3, 5, and 8 were isolated from marine macroalgae for the first time.


Nurse roles should not be at the 'whim of the political class'.
If assistant practitioners are doing the work proposed for nursing associates already then fine, but let us formalise who is responsible for what, right here and now.


Financial incentives enhance adaptation to a sensorimotor transformation.
Adaptation to sensorimotor transformations has received much attention in recent years. However, the role of motivation and its relation to the implicit and explicit processes underlying adaptation has been neglected thus far. Here, we examine the influence of extrinsic motivation on adaptation to a visuomotor rotation by way of providing financial incentives for accurate movements. Participants in the experimental group "bonus" received a defined amount of money for high end-point accuracy in a visuomotor rotation task; participants in the control group "no bonus" did not receive a financial incentive. Results showed better overall adaptation to the visuomotor transformation in participants who were extrinsically motivated. However, there was no beneficial effect of financial incentives on the implicit component, as assessed by the after-effects, and on separately assessed explicit knowledge. These findings suggest that the positive influence of financial incentives on adaptation is due to a component which cannot be measured by after-effects or by our test of explicit knowledge. A likely candidate is model-free learning based on reward-prediction errors, which could be enhanced by the financial bonuses.


Correlation between minimal secretory capacity of pancreatic beta-cells and stability of diabetic control.
The significance of the minimal secretory capacity of pancreatic beta-cells for the stability of the plasma glucose level was studied in 20 patients with insulin-dependent diabetes mellitus. Changes in plasma concentrations of major counterregulatory hormones in response to hypoglycemia were also investigated in these patients to clarify their contribution to diabetic brittleness. beta-Cell function was evaluated on the basis of elevation of plasma C-peptide immunoreactivity (CPR) during the intravenous glucagon test with a highly sensitive assay for plasma CPR that could detect as little as 0.03 ng/ml. After stimulation with glucagon, a significant increase in plasma CPR was observed in 10 of the patients whose beta-cell function had been evaluated as completely depleted by a conventional assay for plasma CPR. A clear inverse correlation was found between the secretory capacity of pancreatic beta-cells measured in this way and the degree of glycemic instability (r = -.74, P less than .01). Infusion of insulin at a rate of 0.15 U.kg-1.h-1 for 60 min caused a continuous decrease in the plasma glucose level, resulting in neuroglycopenia in 7 of the 10 CPR nonresponders but only 2 of the CPR responders. During insulin-induced hypoglycemia, plasma glucagon immunoreactivity did not increase in the CPR nonresponders but increased significantly in the CPR responders. A positive correlation was found between the minimal residual beta-cell capacity and the responsiveness of alpha-cells to hypoglycemia (r = .65, P less than .01).(ABSTRACT TRUNCATED AT 250 WORDS)


Vitamin B₁₂ deficiency-induced increase of osteoclastic bone resorption caused by abnormal renal resorption of inorganic phosphorus via Napi2a.
Vitamin B12 deficiency is a risk factor for bone disorders via mechanisms not fully understood. In this study, an increase in serum inorganic phosphorus (Pi) concentrations was associated with a vitamin B12 deficiency. Napi2a, a renal cotransporter for Pi reabsorption, accumulated on plasma membranes in a vitamin B12 deficiency suggests that vitamin B12 plays an important role in Pi homeostasis.


Amarogentin has protective effects against sepsis-induced brain injury via modulating the AMPK/SIRT1/NF-κB pathway.
Amarogentin (AMA), a secoiridoid glycoside that is mainly derived from SwertiaandGentiana roots, has been confirmed to exhibit antioxidative, tumor-suppressive and anti-diabetic properties. This research intends to investigate the protective effect of AMA against sepsis-induced brain injury and its mechanism. NSC-34 and HT22 cells were treated with lipopolysaccharide (LPS) to induce an in-vitro sepsis model and then treated with varying concentrations (1, 5, 10 µM) of AMA. Cell proliferation and apoptosis were evaluated. The intensity of inflammation and oxidative stress were assessed by different methods. The AMPK/SIRT1/NF-κB pathway expression was determined by WB. An in-vitro sepsis model was set up with cecal ligation and puncture (CLP) in adult C57/BL6J mice, and different concentrations (25, 50, 100 mg/kg) of AMA were applied for treatment. Neurological function was evaluated using the modified neurological severity scores (mNSS), and the brain tissue damage was measured using hematoxylin-eosin (H&E) staining and Nissl staining. Tissue apoptosis was tested using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. Then, the AMPK inhibitor Compound C (CC) was administered to confirm AMA-mediated mechanism. Our finding illustrated that AMA mitigated LPS-induced neuronal damage, inflammation and oxidative stress, activated the AMPK/SIRT1 pathway and choked NF-κB phosphorylation. Furthermore, AMA improved neurological functions of sepsis mice by reliving neuroinflammation and oxidative stress. Inhibition of AMPK attenuated the protective effect of AMA on neurons or the mice's brain tissues. In conclusion, AMA protected against sepsis-induced brain injury by modulating the AMPK/SIRT1/NF-κB pathway.


[Extra dural arachnoid cyst: case report].
Extradural arachnoid cysts are uncommon expanding lesions in the spinal canal, which may communicate with the subarachnoid space. Usually located in the lower thoracic spine, they may cause symptoms by compressing the spinal cord or nerve roots. We report a case of an extradural thoracic arachnoid cyst revealed by progressive spinal cord compression. CT myelography and MRI enabled diagnosis. Rapid neurological improvement was observed after surgical resection.


IL12B polymorphism and type 1 diabetes in the Italian population: a case-control study.
A polymorphism in the interleukin 12B gene was recently reported to be strongly associated with type 1 diabetes in 422 Australian and British families. We analyzed the same polymorphism in 470 Italian type 1 diabetic patients and 544 matched control subjects and found no evidence of association with the disease.


Possible cause of excess sludge reduction in an oxic-settling-anaerobic activated sludge process (OSA process).
Modification of a conventional activated sludge process by inserting a sludge holding tank in a sludge return line forms an oxic-settling-anaerobic (OSA) process that may provide a cost-effective way to reduce excess sludge production in activated sludge processes. In this paper we systematically evaluate the following possible scenarios that may explain the reduction of excess sludge in the OSA process: (i). energy uncoupling, (ii). domination of slow growers, (iii). soluble microbial products (SMPs) effect and (iv). sludge decay in the sludge holding tank under a low oxidation-reduction potential (ORP) condition. Results show that only the final scenario may reasonably explain this reduction. It has also been found that the sludge decay process in the sludge holding tank may involve the reduction of the cell mass.


Comparison between the role of discontinuities in cardiac conduction and in a one-dimensional hardware model.
In real electrophysiological experiments, irregularities in the extracellular excitation spread are believed to depend on cardiac tissue microstructure. An electronic hardware model was developed to analyze this dependence by placing some inhomogeneities (slow propagation areas) in the medium. The position of such inhomogeneities is correlated with abnormal delays and irregularities measured in signal propagation.


Visual function and performance with blue-light blocking filters in age-related macular degeneration.
Some dispute has occurred over the use of blue-light-attenuating intraocular lenses in age-related macular degeneration (AMD), as they may reduce scotopic vision. This study aimed to determine if a blue blocking filter would affect performance during eye-hand coordination and mobility tasks in scotopic illumination, psychophysically measured scotopic sensitivity or colour discrimination in AMD patients. Scotopic measures performed with and without a blue-attenuating filter included a mobility obstacle course, manipulation of cylindrical blocks and a psychophysical dark-adapted full-field flash test. A navy and blue sock colour sorting task evaluated photopic colour discrimination. Subjects were 22 bilateral pseudophakes with early AMD and visual acuity >6/24. On average with the filter, there was a 13% increase in time during the block test. The differences in time and number of bumps with versus without the filter were not significant for the mobility course. Performance with and without the filter was well correlated for the blocks (r = 0.70), flash test (r = 0.83) and mobility (r = 0.66), and the regression slopes were not significantly different from unity. 77% of subjects misidentified at least one navy sock as black with the filter compared with 9% without, with a significant increase in such misidentifications with the filter. The difference in scotopic visual function or performance with versus without a blue-blocking filter most likely does not produce a clinically significant effect or risk; however, detection of navy colour may be impaired.


Effects of normobaric hyperoxia on hemodynamics and O2 utilization in conscious dogs.
Normobaric hyperoxia decreased resting whole-body O2 consumption in conscious dogs by equal (Fick) contributions from decreases in cardiac output and in the arterial-venous difference in O2 content. The decrease in O2 consumption was fully developed by 20 min, was maintained for at least 1 h, and was both reversible and reproducible. Hyperoxia also decreased heart rate, right and left ventricular work rates (and therefore, presumably myocardial O2 consumption), and pulmonary vascular resistance; and increased systemic vascular resistance and right atrial pressure. Paradoxically, hyperoxia did not change O2 delivery. This latter observation together with the decrease in O2 consumption produced a unique vertical orientation to the O2 consumption-delivery relationship induced by hyperoxia. It is concluded that hyperoxia may decrease metabolic rate and substantially alter hemodynamics, which may have important implications for understanding the metabolic regulation of oxygen utilization and for the medical and nonmedical uses of oxygen.


A trophic transfer study: accumulation of multi-walled carbon nanotubes associated to green algae in water flea Daphnia magna.
Carbon nanotubes (CNT) are promising nanomaterials in modern nanotechnology and their use in many different applications leads to an inevitable release into the aquatic environment. In this study, we quantified trophic transfer of weathered multi-walled carbon nanotubes (wMWCNT) from green algae to primary consumer Daphnia magna in a concentration of 100 μg L-1 using radioactive labeling of the carbon backbone (14C-wMWCNT). Trophic transfer of wMWCNT was compared to the uptake by daphnids exposed to nanomaterials in the water phase without algae. Due to the rather long observed CNT sedimentation times (DT) from the water phase (DT50: 3.9 days (d), DT90: 12.8 d) wMWCNT interact with aquatic organisms and associated to the green algae Chlamydomonas reinhardtii and Raphidocelis subcapitata. After the exposition of algae, the nanotubes accumulated to a maximum of 1.6 ± 0.4 μg 14C-wMWCNT mg-1 dry weight-1 (dw-1) and 0.7 ± 0.3 μg 14C-wMWCNT mg-1 dw-1 after 24 h and 48 h, respectively. To study trophic transfer, R. subcapitata was loaded with 14C-wMWCNT and subsequently fed to D. magna. A maximum body burden of 0.07 ± 0.01 μg 14C-wMWCNT mg-1 dw-1 and 7.1 ± 1.5 μg 14C-wMWCNT mg-1 dw-1 for D. magna after trophic transfer and waterborne exposure was measured, respectively, indicating no CNT accumulation after short-term exposure via trophic transfer. Additionally, the animals eliminated nanomaterials from their guts, while feeding algae facilitated their excretion. Further, accumulation of 14C-wMWCNT in a growing population of D. magna revealed a maximum uptake of 0.7 ± 0.2 μg mg-1 dw-1. Therefore, the calculated bioaccumulation factor (BAF) after 28 d of 6700 ± 2900 L kg-1 is above the limit that indicates a chemical is bioaccumulative in the European Union Regulation REACH. Although wMWCNT did not bioaccumulate in neonate D. magna after trophic transfer, wMWCNT enriched in a 28 d growing D. magna population regardless of daily feeding, which increases the risk of CNT accumulation along the aquatic food chain.


The versatile roles of lumican in eye diseases: A review.
Lumican is a keratan sulfate proteoglycan that belongs to the small leucine-rich proteoglycan family. Research has lifted the veil on the versatile roles of lumican in the pathogenesis of eye diseases. Lumican has pivotal roles in the maintenance of physiological tissue homogenesis and is often upregulated in pathological conditions, e.g., fibrosis, scar tissue formation in injured tissues, persistent inflammatory responses and immune anomaly, etc. Herein, we will review literature regarding the role of lumican in pathogenesis of inherited congenital and acquired eye diseases, e.g., cornea dystrophy, cataract, glaucoma and chorioretinal diseases, etc.


A room with a green view: the importance of nearby nature for mental health during the COVID-19 pandemic.
The COVID-19 pandemic and its global response have resulted in unprecedented and rapid changes to most people's day-to-day lives. To slow the spread of the virus, governments have implemented the practice of physical distancing ("social distancing"), which includes isolation within the home with limited time spent outdoors. During this extraordinary time, nature around the home may play a key role in mitigating against adverse mental health outcomes due to the pandemic and the measures taken to address it. To assess whether this is the case, we conducted an online questionnaire survey (n = 3,000) in Tokyo, Japan, to quantify the association between five mental health outcomes (depression, life satisfaction, subjective happiness, self-esteem, and loneliness) and two measures of nature experiences (frequency of greenspace use and green view through windows from home). Accounting for sociodemographic and lifestyle variables, we found that the frequency of greenspace use and the existence of green window views from within the home was associated with increased levels of self-esteem, life satisfaction, and subjective happiness and decreased levels of depression, anxiety, and loneliness. Our findings suggest that a regular dose of nature can contribute to the improvement of a wide range of mental health outcomes. With the recent escalation in the prevalence of mental health disorders, and the possible negative impacts of the COVID-19 pandemic on public mental health, our findings have major implications for policy, suggesting that urban nature has great potential to be used as a "nature-based solution" for improved public health.


Cytogeography of Ixeris nakazonei (Asteraceae, Lactuceae) in the Ryukyu Archipelago of Japan and Taiwan.
The cytogeographical structures of Ixeris nakazonei, a putative hybrid between I. debilis (6x) and I. repens (2x), were investigated in the Ryukyu Archipelago and Taiwan. In the Ryukyus, I. debilis occurs on Miyakojima Island of the southern Ryukyus and northward, while I. repens occurs on all islands except for Iriomotejima and Yonagunijima Islands. I. nakazonei, comprises six polyploid cytotypes, 3x, 4x, 5x, 6x, 7x and 8x, based on x=8. Four cytotypes from 3x to 6x occur in the central Ryukyus, while four cytotypes from 5x to 8x occur in the southern Ryukyus. The higher polyploids of I. nakazonei tend to be distributed in the more southerly area. Tetraploids of I. nakazonei always co-occur with I. debilis and I. repens, supporting the hybrid origin of this cytotype. Considering the chromosome number, octoploids, which predominate in the southern Ryukyus and Taiwan, may have derived directly from hybridization between I. debilis and I. repens. Odd-numbered polyploids of I. nakazonei, 3x, 5x and 7x, are relatively rare. Their chromosome numbers indicate that triploids and heptaploids are hybrids between the tetraploid of I. nakazonei and I. repens, and between the octoploid of I. nakazonei and I. debilis, respectively. Pentaploids of I. nakazonei in the central and southern Ryukyus are, respectively, hybrids between the tetraploid of I. nakazonei and I. debilis and between the octoploid of I. nakazonei and I. repens, indicating that pentaploids of I. nakazonei have at least two independent origins.


Detection and Analysis of West Nile Virus Structural Protein Genes in Animal or Bird Samples.
West Nile virus (WNV) is an important zoonotic pathogen, which is detected mainly by identification of its RNA using PCR. Genetic differentiation between WNV lineages is usually performed by complete genome sequencing, which is not available in many research and diagnostic laboratories. In this chapter, we describe a protocol for detection and analysis of WNV samples by sequencing the entire region of their structural genes capsid (C), preM/membrane, and envelope. The primary step is the detection of WNV RNA by quantitative PCR of the NS2A gene or the C gene regions. Next, the entire region containing the structural protein genes is amplified by PCR. The primary PCR product is then amplified again in parallel reactions, and these secondary PCR products are sequenced. Finally, bioinformatic analysis enables detection of mutations and classification of the samples of interest. This protocol is designed to be used by any laboratory equipped for endpoint and quantitative PCR. The sequencing can be performed either in-house or outsourced to a third-party service provider. This protocol may therefore be useful for rapid and affordable classification of WNV samples, obviating the need for complete genome sequencing.


Automatic initialization of an active shape model of the prostate.
In this work is reported a new method for automatic segmentation of the boundary of the prostate, in transurethral ultrasound images. The scheme is based on a robust automatic initialization of an active shape model (ASM) of the prostate, which is subsequently fitted to the boundary of the gland. The initialization of the ASM is based on pixel classification to estimate the prostate region in an ultrasound image, followed by automatic adjustment - using a multipopulation genetic algorithm (MPGA) - of the initial pose of the ASM to the binary image produced by the classifier. The initial pose is next adjusted to the gray level ultrasound image, using the MPGA. After automatic initialization, the ASM is adjusted to the gray level ultrasound image to produce the final prostate contour. The method provides fast and robust segmentation of the prostate boundary. Validation results on 22 ultrasound images are reported with 1.74 mm of mean boundary error and an estimated processing time of 66 per image. Our automatic initialization method can be applied with the ASMs of different organs in various imaging modalities.


Prediction of recurrence following hepatectomy in patients with hepatitis C virus infection-related hepatocellular carcinoma who achieved a sustained virological response.
The risk of hepatitis C virus infection-related hepatocellular carcinoma (HCC) is lower, with a better prognosis, in patients who achieve a sustained virological response (SVR) than in those who do not. We aimed to identify risk factors of post-hepatectomy HCC recurrence in patients who achieved a SVR. This retrospective study included 349 HCC patients who underwent an initial radical hepatectomy at our institution between January 2005 and December 2014. Sixty-eight patients had achieved a SVR (the SVR group) and 281 patients had not (the non-SVR group). Clinical characteristics and long-term outcomes were compared between the two groups. Univariate and multivariate analyses identified variables associated with recurrence-free survival in the SVR group. Post-hepatectomy overall and recurrence-free survival rates were significantly higher in the SVR group than the non-SVR group (P < 0.01 and <0.05, respectively). Univariate analysis of post-hepatectomy recurrence-free survival in the SVR group revealed multiple significant factors: aspartate aminotransferase, 25 IU/L or more (P = 0.01); indocyanine green retention rate at 15 min, 20.0% or less (P < 0.05); hepatic vascular invasion (P < 0.05); and an interval of months or less between achieving a SVR and hepatectomy (P < 0.01). Multivariate analysis confirmed an interval of 30 months or less between achieving a SVR and hepatectomy as an independent prognostic factor of recurrence-free survival (hazard ratio, 2.30; 95.0% confidence interval, 1.04-5.13; P < 0.05). The interval between achieving a SVR and hepatectomy is an important predictor of recurrence in hepatitis C virus infection-related HCC patients who achieved a SVR.


Highly selective and sensitive chemosensor for Al(III) based on isoquinoline Schiff base.
As a colorimetric and fluorescent turn-on sensor to Al3+, N'-(2-hydroxybenzylidene)isoquinoline-3-carbohydrazide (HL) has been easily synthesized. The fluorescence intensity increases by 273 times in the presence of Al3+ at 458 nm. Meanwhile, the experiment data indicate that the limit of detection for Al3+ is 1.11 × 10-9 M. Remarkably, the blue fluorescence signal of HL-Al3+ could be specially observed by the naked eye under UV light and is significantly different from those of other metal ions. Fluorescence switch based on the control of Al3+ and EDTA proved HL could act as a reversible chemosensor. According to ESI-MS result and the Job's plots, the 2:1 coordination complex formed by HL and Al3+ could be produced. Density functional theory calculations were performed to illustrate the structures of HL and complex. The cell imaging experiment indicates that HL can be applied for monitoring intracellular Al3+ levels in cells.


Sex-Related Disparities in Acute Limb Ischemia Treatment Outcomes.
Although a substantial impetus behind disparities research in healthcare exists, those that are sex-related within vascular surgery outcomes are largely unexplored. Consequently, published guidelines lack specificity when it comes to treating male and female patients with vascular disease. Disparities related to patients suffering from chronic limb-threatening ischemia have been broached, although no extensive studies assessing disparities in acute limb ischemia treatment outcomes have come to the forefront. In this study, our aim is to identify and quantify sex-related disparities as they pertain to interventions for acute limb ischemia. Using the TriNetX global research network, we conducted a multicenter query across 48 healthcare organizations spanning 5 countries for patients treated for acute limb ischemia. We determined the number of male and female patients that received one of the following interventions: open revascularization, percutaneous mechanical thrombectomy, or catheter-directed thrombolysis and/or adjunctive endovascular procedures. Propensity score matching was performed to account for comorbidities. Risk of adverse outcomes within 30 days was calculated for each sex, including reintervention, major amputation, and death. Risk of adverse outcomes was then compared between treatment groups of the same sex and between sexes. Type-I errors were reduced through utilization of the Holm-Bonferroni method to correct P values. Within our study, we noted several important findings. Females were more likely to receive catheter-directed thrombolysis and/or adjunctive endovascular procedures (P = 0.001) than males. There were no significant differences in the rates of open revascularization or percutaneous mechanical thrombectomy between males and females. Overall, females were more likely to die within 30 days (P < 0.0001) and greater number of males required reintervention within 30 days (P < 0.0001). Analyzing outcomes within individual treatment groups, females undergoing open revascularization or catheter-directed thrombolysis and/or adjunctive endovascular intervention demonstrated a profound increase in mortality within 30 days of intervention (P = 0.0072 and P = 0.0206, respectively), but these differences were not reflected in the percutaneous mechanical thrombectomy group. Limb salvage rates in females were higher than males overall although there were no significant sex differences within any treatment groups specifically. In conclusion, there was a significantly higher risk of death in females across all treatment groups in the studied timeframe. Limb salvage rates were higher for females in the open revascularization (OR) treatment group, while males were more likely to require a reintervention across all treatment groups. By evaluating these disparities, we can provide greater insight into personalized treatment for patients presenting with acute limb ischemia.


High-performance K-ion half/full batteries with superb rate capability and cycle stability.
SignificanceThe present work might be significant for exploring advanced K-ion batteries with superb rate capability and cycle stability toward practical applications. The as-assembled K-ion half cell exhibits an excellent rate capability of 428 mA h g-1 at 100 mA g-1 and a high reversible specific capacity of 330 mA h g-1 with 120% specific capacity retention after 2,000 cycles at 2,000 mA g-1, which is the best among those based on carbon materials. The as-constructed full cell delivers 98% specific capacity retention over 750 cycles at 500 mA g-1, superior to most of those based on carbon materials that have been reported thus far.


Oligomeric intermediates in amyloid formation: structure determination and mechanisms of toxicity.
Oligomeric intermediates are non-fibrillar polypeptide assemblies that occur during amyloid fibril formation and that are thought to underlie the aetiology of amyloid diseases, such as Alzheimer's disease, Parkinson's disease and Huntington's disease. Focusing primarily on the oligomeric states formed from Alzheimer's disease β-amyloid (Aβ) peptide, this review will make references to other polypeptide systems, highlighting common principles or sequence-specific differences. The covered topics include the structural properties and polymorphism of oligomers, the biophysical mechanism of peptide self-assembly and its role for pathogenicity in amyloid disease. Oligomer-dependent toxicity mechanisms will be explained along with recently emerging possibilities of interference.


Cellular and humoral immune responses to chimeric EGFP-pseudocapsids derived from the mouse polyomavirus after their intranasal administration.
Mouse polyomavirus (MPyV) VP1-pseudocapsids carrying enhanced green fluorescent protein (EGFP-VLPs) were used for intranasal immunization of mice. EGFP-VLPs induced strong anti-VP1 but not anti-EGFP antibody production. In vitro restimulation with antigen-pulsed bone marrow-derived dendritic cells (BMDCs) induced remarkable T-cell proliferative response specific for both VP1 and EGFP antigen and IL-2 and IFN-gamma production. Surprisingly, no specific cytotoxic activities against VP1 and EGFP proteins were detected. After intranasal administration of EGFP-VLPs, as well as after polyomavirus infection, a moderate reduction of CD4(+)CD25(+)Foxp3(+) T cells was observed in spleens but not in lymph nodes and peripheral blood, suggesting that both MPyV virions and pseudocapsids are able to induce changes in distribution of regulatory T cells. Treatment of EGFP-VLPs pulsed BMDCs with inhibitors of endosomal acidification proved that presentation of peptides on MHCgp class II is dependent on acidic endosomal environment. Substantial decrease of CD4-specific T-cell proliferation in the presence of proteasome inhibitor suggests that MHCgp class II might load VPL-derived peptides processed by proteasomes. Thus, polyomavirus derived VLPs appear to be promising delivery and adjuvant vehicles for therapeutic proteins.


Update on feline calicivirus: new trends.
In addition to being important upper respiratory tract pathogens of cats, FCVs are increasingly reported as a cause of a highly contagious febrile hemorrhagic syndrome. Strains causing this syndrome are genetically different from the vaccine strain and other nonhemorrhagic FCV isolates. They apparently differ from one outbreak to another. The syndrome is characterized variably by fever; cutaneous edema and ulcerative dermatitis; upper respiratory tract signs; anorexia; occasionally icterus, vomiting, and diarrhea; and a mortality that approaches 50%. Adult cats tend to be more severely affected than kittens, and vaccination does not appear to have a significant protective effect. Rapid recognition of the disease through identification of clinical signs and appropriate testing, followed by strict institution of disinfection, isolation, and quarantine measures, are essential to prevent widespread mortality resulting from the infection.


Patterns of intra- and interspecific association of gall-forming sawflies in relation to shoot size on their willow host plant.
Four species of gall-forming sawflies were each frequently found to have clumped distributions among shoots within their willow host plant across four sites and three years. When all species were considered together by clone, year, and site, species showed independence of distribution among shoots two thirds of the time and showed positive covariance one third of the time. When pairs of species were considered separately, but clones were combined within sites and years, 60% of the chi-square tests of association were significant. All but one of the significant tests showed positive associations between pairs of species. The stem galler was positively associated with the leaf folder at all sites in all years, and the petiole galler was positively associated with the stem galler and leaf folder for most year by site combinations. When species paris co-occurred on shoots they were usually found at the same or higher density as when found alone on shoots. Only 2 of 100 tests showed a depressed density of a species when co-occurring on shoots with heterospecifics.All sawfly species were found on shoots that were significantly larger (mean node number) than on shoots without sawflies, and species responded to shoot size variation similarly. Sizes of shoots occupied by heterospecific species combinations were usually significantly larger than shoots with only conspecifics, for all species. These data supported the hypothesis that similar species' responses to within-plant variation would lead to positive rather than negative or random species associations. The data do not support the hypothesis that interspecific competition was important in determining shoot choice or species density.


Diversity synthesis using the complimentary reactivity of rhodium(II)- and palladium(II)-catalyzed reactions.
Rhodium(II)-catalyzed reactions of aryldiazoacetates can be conducted in the presence of iodide, triflate, organoboron, and organostannane functionality, resulting in the formation of a variety of cyclopropanes or C-H insertion products with high stereoselectivity. The combination of the rhodium(II)-catalyzed reaction with a subsequent palladium(II)-catalyzed Suzuki coupling offers a novel strategy for diversity synthesis.


Orthotopic cardiac transplantation for the treatment of progressive heart failure caused by idiopathic giant cell myocarditis.
We describe a patient with idiopathic giant cell myocarditis who underwent orthotopic cardiac transplantation. On triple immunosuppressive therapy including prednisone, cyclosporine and mycophenolate, the patient has not had recurrence of idiopathic giant cell myocarditis in the transplanted heart in 48 months of follow-up.


Comparison of separate hydrolysis and fermentation and simultaneous saccharification and fermentation processes for ethanol production from wheat straw by recombinant Escherichia coli strain FBR5.
Ethanol production by recombinant Escherichia coli strain FBR5 from dilute acid pretreated wheat straw (WS) by separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) was studied. The yield of total sugars from dilute acid (0.5% H(2)SO(4)) pretreated (160 °C, 10 min) and enzymatically saccharified (pH 5.0, 45 °C, 72 h) WS (86 g/l) was 50.0 ± 1.4 g/l. The hydrolyzate contained 1,184 ± 19 mg furfural and 161 ± 1 mg hydroxymethyl furfural per liter. The recombinant E. coli FBR5 could not grow at all at pH controlled at 4.5 to 6.5 in the non-abated wheat straw hydrolyzate (WSH) at 35 °C. However, it produced 21.9 ± 0.3 g ethanol from non-abated WSH (total sugars, 44.1 ± 0.4 g/l) in 90 h including the lag time of 24 h at controlled pH 7.0 and 35 °C. The bioabatement of WS was performed by growing Coniochaeta ligniaria NRRL 30616 in the liquid portion of the pretreated WS aerobically at pH 6.5 and 30 °C for 15 h. The bacterium produced 21.6 ± 0.5 g ethanol per liter in 40 h from the bioabated enzymatically saccharified WSH (total sugars, 44.1 ± 0.4 g) at pH 6.0. It produced 24.9 ± 0.3 g ethanol in 96 h and 26.7 ± 0.0 g ethanol in 72 h per liter from bioabated WSH by batch SSF and fed-batch SSF, respectively. SSF offered a distinct advantage over SHF with respect to reducing total time required to produce ethanol from the bioabated WS. Also, fed-batch SSF performed better than the batch SSF with respect to shortening the time requirement and increase in ethanol yield.


[An experimental study of small-caliber tissue engineering vessels with acellular matrix].
To observe the mechanical properties of the prefabricated connective tissue tube as blood vessel substitute and its changes after implantation at the femoral artery. The acellular matrix tube of 8-12 cm in length with a silicone rod inside it was implanted into dog peritoneal cavity. 3 weeks later, a new formed tube around the silicone rod was transferred to the femoral artery as blood vessel substitute. The mechanical properties and histological examination of the blood vessel substitute were assessed and compared to those of the carotid artery and vein. 6 months after transfer, the patency of the blood vessels substitute was observed. The histological change was studied by light microscopy, scanning and transmitting electron microscopy. (1) The mechanical properties of blood vessel substitute was not as strong as artery, but better than the vein. (2) There were elastic and collagen fibers with many fibroblasts around the tube wall, but few mesothelial cells around the inner wall. All of the blood vessel substitutes (n = 6) were found to keep patency and the structure of the blood vessels substitutes became similar to femoral artery 6 months after they had been grafted to the femoral artery. These results suggest that tissue engineering in vivo is a good approach to construct vessels substitute. The tissue tubes made in dog's peritoneal cavity have good condition when it is used as a blood vessel substitutes.


Electrocardiogram interpretation in college athletes: Local institution versus sports cardiology center interpretation.
Electrocardiogram (ECG) screening in athletes enhances the detection of conditions associated with sudden cardiac death (SCD), but concerns remain for false positive results when conducted outside of specialized centers. This study compared ECG interpretation in college athletes between local physicians and a sports cardiology center (SCC). Screening ECGs in athletes from eight Pacific-12 Conference institutions performed between 2010 and 2016 were included. Local interpretation was compared to SCC interpretation using both the Seattle Criteria (SCC-SC) and the International Criteria (SCC-IC). A total of 2445 athlete ECGs (mean age 18.5 years; 57.1% male; 63.2% Caucasian and 15.3% African American) were reviewed. The proportion of ECGs classified as abnormal was similar between local and SCC-SC interpretation (3.5% vs. 3.4%, respectively; p = .94), but was lower by SCC-IC interpretation (1.5%, p < .001). ECG abnormalities interpreted as normal by local physicians but as abnormal by SCC-SC (n = 33) and SCC-IC (n = 16) standards included: pathological Q waves (n = 15 SCC-SC; n = 3 SCC-IC), T-wave inversions (n = 8 both), and ST-depressions (n = 3 both). There was a 97.5% ECG interpretation agreement and substantial interobserver reliability (k = 0.611, p < .001) between local and SCC-SC interpretation in athletes screened starting one year after publication of the Seattle Criteria (n = 1388). Both local and SCC physicians correctly identified six abnormal ECGs associated with conditions at risk of SCD. ECG interpretation by local physicians at college universities had similar accuracy compared to a specialized SCC with a low overall abnormal rate, similar sensitivity, and substantial interobserver reliability. Uniform application of current ECG interpretation standards is recommended to further improve accuracy.


Asymmetric syntheses of 8-oxabicyclo[3,2,1]octane and 11-oxatricyclo[184.108.40.206]undecane from glycals.
Herein, we describe an efficient method to prepare enantiomerically pure 8-oxabicyclo[3.2.1]octanes via gold(i)-catalyzed tandem 1,3-acyloxy migration/Ferrier rearrangement of glycal derived 1,6-enyne bearing propargylic carboxylates. The resultant compounds could then undergo interrupted Nazarov cyclization to afford diastereomerically pure 11-oxatricyclo[126.96.36.199]undecanes.


A Patch Testing Initiative for the Investigation of Allergic Contact Dermatitis in a UK Allergy Practice: A Retrospective Study.
Patch testing is the gold standard diagnostic tool for investigating allergic contact dermatitis (ACD). In the United Kingdom, patch testing has been historically confined to the dermatologist's office. Furthermore, detailed studies on patch testing by allergists are significantly underrepresented at the international level. The objective of this study was to undertake a comprehensive evaluation of a patch testing initiative from an allergy practice; we report on various patient characteristics, prevalence and relevance data, in addition to immediate hypersensitivity testing. We undertook a retrospective analysis of 156 patients suspected of having contact dermatitis seen in our UK allergy practice between October 2016 and April 2018. Of the 156 patients patch tested (mean age 36.9 years, female 88%, white ethnicity 71.8%, atopy 68.6%), ACD was diagnosed overall in 49% of the cohort and ACD of current relevance was assigned to 31%. Our extended British standard series alone detected the responsible allergen in 87% of patients, and the remaining 13% were detected from supplementary or own material testing alone. Most prevalent contact allergens were nickel (28.2%), p-phenylenediamine (8.3%), cobalt (8.3%), methylisothiazolinone (5.8%), and hydroperoxides of linalool (4.5%) and limonene (4.5%). A history of occupationally related dermatitis (P = .004) and initial (pretest) diagnosis of ACD (P < .001) were both significantly associated with relevant positive patch test reactions (atopy status was not associated P > .05). ACD was detected in almost 50% of assessed patients, and we highlight the importance of assessing relevance. Hydroperoxides of limonene and linalool are notable additions to the prevalence data. Patch testing should be incorporated into more allergy practices, although availability of training is a limiting factor.


Cystathionine in rat brain: catabolism in vivo.
The content of cystathionine was measured in 35 rat brains; the range was 10-120 nmol/g wet weight and thus the variability of cystathionine content in rat brain was emphasized. The regional distribution of cystathionine was also determined: the highest level was found in cerebellum; the lowest level was observed in the white and gray matter of the hemispheres. These results are different from those obtained in other species. The radioactive metabolites formed from L-(35S)cystathionine injected intracisternally were measured in brains of rats killed at the following times after injection: 0.25, 1, 2, 4,6, 9, 16, and 27 hr. The radioactivity was found both in the proteins and in the acid-soluble fraction. In the acid-soluble fraction the radioactivity was found in various ninhydrin-reacting compounds: [cysteic cysteine sulfinic] acid, taurine, reduced and oxidized glutathione, cystine, cystathionine, and a compound tentatively identified as the mixed disulfide of cysteine and glutathione. The radioactivity of cystathionine decreased exponentially between the 1st and the 27th hour after injection and its half-life was estimated to be about 5 hr. The radioactivity in the other ninhydrin-reacting compounds increased until the 9th hour after injection, then decreased. Half of this radioactivity was present in reduced glutathione, the rest being shared equally between: [cysteic cysteine sulfinic] acid, taurine, and the mixed disulfide. It is worthwhile to note that the radioactivity in the cystine fraction was always very low.


Acacetin and chrysin, two polyphenolic compounds, alleviate telomeric position effect in human cells.
We took advantage of the ability of human telomeres to silence neighboring genes (telomere position effect or TPE) to design a high-throughput screening assay for drugs altering telomeres. We identified, for the first time, that two dietary flavones, acacetin and chrysin, are able to specifically alleviate TPE in human cells. We further investigated their influence on telomere integrity and showed that both drugs drastically deprotect telomeres against DNA damage response. However, telomere deprotection triggered by shelterin dysfunction does not affect TPE, indicating that acacetin and chrysin target several functions of telomeres. These results show that TPE-based screening assays represent valuable methods to discover new compounds targeting telomeres.Molecular Therapy-Nucleic Acids (2013) 2, e116; doi:10.1038/mtna.2013.42; published online 20 August 2013.


Development of highly sensitive and selective antibodies for the detection of the explosive pentaerythritol tetranitrate (PETN) by bioisosteric replacement.
An improved antibody against the explosive pentaerythritol tetranitrate (PETN) was developed. The immunogen was designed by the concept of bioisosteric replacement, which led to an excellent polyclonal antibody with extreme selectivity and immunoassays of very good sensitivity. Compounds such as nitroglycerine, 2,4,6-trinitrotoluene, 1,3,5-trinitrobenzene, hexogen (RDX), 2,4,6-trinitroaniline, 1,3-dinitrobenzene, octogen (HMX), triacetone triperoxide, ammonium nitrate, 2,4,6-trinitrophenol and nitrobenzene were tested for potential cross-reactivity. The detection limit of a competitive enzyme-linked immunosorbent assay was determined to be around 0.5 µg/l. The dynamic range of the assay was found to be between 1 and 1000 µg/l, covering a concentration range of three decades. This work shows the successful application of the bioisosteric concept in immunochemistry by exchange of a nitroester to a carbonate diester. The antiserum might be used for the development of quick tests, biosensors, microtitration plate immunoassays, microarrays and other analytical methods for the highly sensitive detection of PETN, an explosive frequently used by terrorists, exploiting the extreme difficulty of its detection.


Electron microscope studies on the intracellular growth of PL-1 phage of Lactobacillus casei.
Ultrathin sections of the cells of Lactobacillus casei infected with or without PL-1 phages were observed by the rapid-freezing and substitution-fixation method. Phage-head-like particles were first observed in the nuclear region. The region was seen more widely dispersed in the cytoplasm than that observed by the conventional chemical fixation method. The features of cells just broken open by the infected phages were observed by the sedimentation method devised by us. The bursting occurred in more than one place in the cells with liberation of progeny phages.


S-nitrosation and thiol switching in the mitochondrion: a new paradigm for cardioprotection in ischaemic preconditioning.
Understanding the molecular mechanisms through which the heart could be protected from ischaemic injury is of major interest and offers a potential route for the development of new therapies. Recently, several studies have uncovered intriguing relationships between nitric oxide-induced protein thiol modifications and the cardioprotected phenotype. In a highly cited, seminal article published in the Biochemical Journal in 2006, Burwell and colleagues addressed this issue and provided direct evidence for S-nitrosation of complex I of the mitochondrial electron transport chain. These authors were the first to show increased S-nitrosation of mitochondrial proteins from hearts subjected to the cardioprotective process known as ischaemic preconditioning. This study has paved the way for further investigations that collectively reveal a potential link between the mitochondrial S-nitrosoproteome and ischaemic preconditioning.


Sleep disturbances increase interleukin-6 production during pregnancy: implications for pregnancy complications.
Pregnant women experience disturbed sleep that varies throughout the gestational period. In clinical studies of nonpregnant cohorts, data link disturbed sleep with increases in inflammatory markers. Emerging evidence has also found associations between increased inflammation and medical morbidity, including various pregnancy complications. The authors have previously shown a correlation between sleep disturbances and serum cytokine levels. They extend this initial observation by evaluating the relationship between sleep during mid and late pregnancy and inflammatory cytokines in both serum and stimulated peripheral blood mononuclear cells. Subjective sleep during pregnancy, described by the Pittsburgh Sleep Quality Index and sleep diaries, and circulating and stimulated measures of interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha were evaluated in 19 pregnant women. The authors found that greater sleep complaints in late pregnancy were associated with both increased circulating and stimulated IL-6 levels. Short sleep duration and poor sleep efficiency in both mid and late pregnancy were associated with higher stimulated levels of IL-6. No relationships were observed for TNF-alpha. These preliminary findings indicate that women who experience sleep disturbances as early as mid gestation are likely to have an increase in inflammation.


The Burn Wheel: a practical guide to fluid resuscitation.
During preparation for mass casualties of burn victims in the Gulf War (1990-91), a 'Battlefield Burns Table' was devised to allow rapid calculation of fluid replacement. By standardizing weights and using only Hartmann's Solution BP, a regimen could be prescribed by the non-specialist without resort to mathematical calculations. By means of a revolving disc, this has now been refined for use in a civilian setting where expert burn advice may not be immediately available.


Discovery of novel indazole-acylsulfonamide hybrids as selective Mcl-1 inhibitors.
Overexpressing myeloid cell leukemia sequence 1 (Mcl-1) protein is an important way to confer the resistance of cancer cells to conventional anti-cancer treatments. Therefore, developing Mcl-1 inhibitors has become an attractive strategy for cancer therapy. In the studies, a series of new indazole-acylsulfonamide hybrids were designed, synthesized and evaluated as potent Mcl-1 inhibitors. Among them, the most potent compound 17 (Ki = 0.43 μM) showed a little better inhibitory activity against Mcl-1 protein than positive control AT-101 (Ki = 0.45 μM). Pleasingly, it displayed > 40-fold selectivity over Bcl-2 (Ki = 18 μM) and Bcl-xL (no activity). Furthermore, compound 17 had good inhibitory activities against PC-3, MDA-MB-231 and K562 cells (IC50 = 12.3, 10.6 and 6.62 μM, respectively) and could effectively induce apoptosis and the activation of caspase-3 in a dose-dependent manner in K562 cells.