Pathogenic aspects of dermatomyositis, polymyositis and overlap myositis.
Inflammatory myopathies (IMs) often have distinct histopathologic features suggesting humorally mediated involvement of the microcirculation in dermatomyositis (DM), including early capillary deposition of the complement C5b-9 membranolytic attack complex (MAC) and secondary ischaemic changes; and CD8 T-cell-mediated and MHC1-restricted autoimmune attack of myofibers in polymyositis (PM) and inclusion body myositis. Novel insights in these specific diseases include emerging evidence that capillary loss involves whole microvascular units in DM, and that regulatory T-cells strongly protect myofibers from experimental autotoxic attack in PM. However, all IMs do not exhibit pathophysiology-relevant histopathologic features of DM or PM. Autoimmune necrotizing myopathies (AINM) occur in the absence of endomysial inflammatory cells and may be specifically associated with anti-SRP autoantibodies. Moreover, IM histopathological features may be scarce, unspecific and overlapping. Therefore, increasing attention is paid to features shared by IMs regardless of their type, relevant to the innate immune response and to non-immune mechanisms. Innate immune responses to myodamage (and/or as yet unknown stimuli), involves release of chemokines, activation of specific Toll-like receptors (TLRs) and complex Th-1, Th-17 and other cytokine interplays; it triggers DC recruitment and maturation, and is associated with type 1 IFN signature (especially in DM where type 1 IFN-producing cells called plasmacytoid DCs are mainly detected). Non-immune mechanisms mainly include endoplasmic reticulum (ER) stress induced in myofibers by up-regulation of MHC-class I antigens (as typically observed in PM with a diffuse pattern and in DM with perifascicular predominance). ER stress may favour autoimmune reactions but may also be associated with myofiber damage and dysfunction in the absence of lymphocytes. Overlap myositis (OM) may be associated with other connective tissue diseases and a variety of autoantibodies, such as those directed against tRNA synthetase. Myositis specific autoantibodies are mainly expressed by regenerating myofibers, that may also express MHC-1 and endogenous ligand-binding TLRs, thus drawing a picture in which the regenerating myofiber plays a central pathophysiologic role.


Synthetic retinoids, retinobenzoic acids, Am80, Am580 and Ch55 regulate morphogenesis in chick limb bud.
The retinobenzoic acids Am80, Am580 and Ch55 are synthetic stable analogs of retinoic acid (RA), and show very strong differentiation-inducing activity in human myelogeneous leukemia cell line HL-60. To examine the effects of these synthetic retinoids on limb pattern formation, AG1-X2 beads containing these retinoids were applied to the anterior margin of stage 19-20 chick wing buds. By implanting the beads with 1 microgram/ml retinoids, normal wings were formed and extra digits 2 or 32 were rarely formed. As the retinoid concentrations increased from 10 micrograms/ml to 100 micrograms/ml duplicated limbs 3234, 43234, 432234, 4334 were progressively produced. At higher concentrations, 1 mg/ml, the wings often truncated, although duplication occurred in some embryos. These synthetic analogs seem to have the same degree of morphogenetic potential as RA, since the activity index of these retinoids was similar to that of RA. Since these synthetic retinoids hardly bind to CRABP (cellular retinoic acid-binding protein), it may be possible that the retinoids and RA may affect limb-pattern formation without the interaction with CRABP. It is known that limb buds cannot develop distal structures when the posterior region including all ZPA (zone of polarizing activity) is removed. When beads containing the above mentioned retinoids were implanted to the anterior margin of wing buds from which the posterior one third region including all ZPA had been removed, distal growth of the wing buds and the formation of digit elements were observed. Some of the wing buds produced a completely reverse digit pattern 432. From these results, we discussed the roles of RA in limb development and pattern formation.


An Experimental Comparison of Techniques: Cognitive Defusion, Cognitive Restructuring, and in-vivo Exposure for Social Anxiety.
One of the primary differences between Cognitive Behavioral Therapy (CBT) and Acceptance and Commitment Therapy (ACT) for anxiety is the approach to managing negative thoughts. CBT focuses on challenging the accuracy of dysfunctional thoughts through cognitive restructuring exercises, whereas ACT attempts to foster acceptance of such thoughts through cognitive defusion exercises. Previous research suggests that both techniques reduce the distress associated with negative thoughts, though questions remain regarding the benefit of these techniques above and beyond exposure to feared stimuli. In the present study, we conducted a brief experimental intervention to examine the utility of cognitive defusion + in-vivo exposure, cognitive restructuring + in-vivo exposure, and in-vivo exposure alone in reducing the impact of negative thoughts in patients with social anxiety disorder. All participants completed a brief public speaking exposure and those in the cognitive conditions received training in the assigned cognitive technique. Participants returned a week later to complete a second exposure task and self-report measures. All three conditions resulted in similar decreases in discomfort related to negative thoughts. ANOVA models failed to find an interaction between change in accuracy or importance and assignment to condition in predicting decreased distress of negative thoughts. These preliminary results suggest that changes in perceived importance and accuracy of negative thoughts may not be the mechanisms by which cognitive defusion and cognitive restructuring affect distress in the short-term.


Safety, pharmacokinetics and pharmacodynamics of HWH486 capsules in healthy adults: A randomized, double-blind, placebo-controlled, phase I dose-escalation study.
HWH486 inhibits Bruton's tyrosine kinase and therefore shows promise as a treatment against rheumatoid arthritis and chronic spontaneous urticaria. This phase I trial assessed tolerability, safety, pharmacokinetics and pharmacodynamics of a single oral dose of HWH486 capsules in healthy adults. A single-center, randomized, double-blind, placebo-controlled, dose-escalation study from 10 to 800 mg was conducted in 96 healthy Chinese adults, of whom 80 received HWH486 and 16 received placebo. A total of 96 subjects were enrolled, and all completed the study. In the HWH486 group, mean Tmax ranged from 1.03 to 2.00 h, and mean T1/2 ranged from 0.85 to 8.67 h across the dose range from 10 to 800 mg. Mean Cmax increased linearly with dose, while mean AUC0-t increased non-linearly. Occupancy of Bruton's tyrosine kinase peaked within 0.50-4.00 h after administration across the dose groups, and the delay until peak occupancy decreased with increasing dose. Twenty-five subjects (31.25 %) in the HWH486 group experienced 35 treatment-emergent adverse events, while four subjects (25.00 %) in the placebo group experienced eight such events. HWH486 is well tolerated and safe in healthy adults, in whom it can strongly bind Bruton's tyrosine kinase. These findings justify clinical studies of HWH486 efficacy against autoimmune diseases.


A comprehensive systematic review of the development process of 104 patient-reported outcomes (PROs) for physical activity in chronically ill and elderly people.
Capturing dimensions of physical activity relevant to patients may provide a unique perspective for clinical studies of chronically ill patients. However, the quality of the development of existing instruments is uncertain. The aim of this systematic review was to assess the development process of patient-reported outcome (PRO) instruments including their initial validation to measure physical activity in chronically ill or elderly patient populations. We conducted a systematic literature search of electronic databases (Medline, Embase, Psychinfo, Cinahl) and hand searches. We included studies describing the original development of fully structured instruments measuring dimensions of physical activity or related constructs in chronically ills or elderly. We broadened the population to elderly because they are likely to share physical activity limitations. At least two reviewers independently conducted title and abstract screening and full text assessment. We evaluated instruments in terms of their aim, items identification and selection, domain development, test-retest reliability, internal consistency, validity and responsiveness. Of the 2542 references from the database search and 89 from the hand search, 103 full texts which covered 104 instruments met our inclusion criteria. For almost half of the instruments the authors clearly described the aim of the instruments before the scales were developed. For item identification, patient input was used in 38% of the instruments and in 32% adaptation of existing scales and/or unsystematic literature searches were the only sources for the generation of items. For item reduction, in 56% of the instruments patient input was used and in 33% the item reduction process was not clearly described. Test-retest reliability was assessed for 61%, validity for 85% and responsiveness to change for 19% of the instruments. Many PRO instruments exist to measure dimensions of physical activity in chronically ill and elderly patient populations, which reflects the relevance of this outcome. However, the development processes often lacked definitions of the instruments' aims and patient input. If PROs for physical activity were to be used in clinical trials more attention needs to be paid to the establishment of content validity through patient input and to the assessment of their evaluative measurement properties.


Central venous access: evolving roles of radiology and other specialties nationally over two decades.
The aim of this study was to evaluate national trends in central venous access (CVA) procedures over 2 decades with regard to changing specialty group roles and places of service. Aggregated claims data for temporary central venous catheter and long-term CVA device (CVAD) procedures were extracted from Medicare Physician/Supplier Procedure Summary Master Files from 1992 through 2011. Central venous catheter and CVAD procedure volumes by specialty group and place of service were studied. Between 1992 and 2011, temporary and long-term CVA placement procedures increased from 638,703 to 808,071 (+27%) and from 76,444 to 316,042 (+313%), respectively. For temporary central venous catheters, radiology (from 0.4% in 1992 to 32.6% in 2011) now exceeds anesthesiology (from 37% to 22%) and surgery (from 30.4% to 11.7%) as the dominant provider group. Surgery continues to dominate in placement and explantation of long-term CVADs (from 80.7% to 50.4% and from 81.6% to 47.7%, respectively), but radiology's share has grown enormously (from 0.7% to 37.6% and from 0.2% to 28.6%). Although volumes remain small (<10% of all procedures), midlevel practitioners have experienced >100-fold growth for most services. The inpatient hospital remains the dominant site for temporary CVA procedures (90.0% in 1992 and 81.2% in 2011), but the placement of long-term CVADs has shifted from the inpatient (from 68.9% to 45.2%) to hospital outpatient (from 26.9% to 44.3%) setting. In all hospital settings combined, radiologists place approximately half of all tunneled catheters and three-quarters all peripherally inserted central catheters. Over the past 2 decades, CVA procedures on Medicare beneficiaries have increased considerably. Radiology is now the dominant overall provider.


Extradural origin of the posteroinferior cerebellar artery: an anatomic study with histological and radiographic correlation.
The posteroinferior cerebellar artery (PICA) usually arises from the intradural segment of the vertebral artery (VA). The extradural origin of the PICA is infrequent. Its preoperatory identification is important in surgical strategy during the exposure of the VA. During an anatomic prosection, the VA was exposed at the craniocervical junction in cadaveric adult specimens. The extradural origin of the PICA was encountered bilaterally in one specimen and on one side in a second specimen. An anatomic study with histological and radiographic correlation was performed. Perforating branches originate from the PICA. They supply the middle and inferior third of the olive and the lateral aspect of the medulla. The PICA has cortical branches that lead to the cerebellum. Injury to the PICA can produce an infarction of these neural structures that can be asymptomatic or cause major neurological deficits. Radiographic results obtained using a lateral projection provided the most reliable delineation of the extradural origin of the PICA. When this artery originates at, or posterior to, the posterior aspect of the occipital condyle, an extradural origin is likely. Bilateral selective vertebral angiography should be performed with special attention to the relationships of PICA origins before any surgical exposure of the VA at the craniocervical junction, unless magnetic resonance angiography provides this information without question. A thorough understanding of the relative dominance of the VAs and PICAs, the location of the PICA origin, and the collateral circulation of the posterior fossa are prerequisites to surgery in this region. The preoperative identification of an extradural PICA is important in planning surgical strategy and in avoiding complications during operations near the foramen magnum.


Small cell carcinoma of the cervix.
Small cell carcinoma of the cervix represents an uncommon variant of cervical cancer with a particularly poor prognosis. Traditionally, the diagnosis was established from routine histopathological sections, but there has been a trend to refer to this tumour as neuroendocrine carcinoma, with a requirement to demonstrate cytoplasmic granules. Five patients are described, who share the clinical features of young age of onset, early metastasis in the presence of apparently low stage disease, early failure of appropriate local treatment, and extreme chemosensitivity (features that are quite distinct from those seen in squamous cell cancer of the cervix). Light microscopy suggested a diagnosis of small cell cancer in all five tumours, but not all showed evidence of neuroendocrine differentiation. It is proposed that the present criteria for the diagnosis of cervical small cell carcinoma are too strict. The diagnosis should rely on the light microscopy of haemotoxylin and eosin sections and the distinctive clinical behaviour. The absence of neuroendocrine differentiation should not exclude the diagnosis, it does not appear to influence the clinical behaviour. The appropriate management of small cell carcinoma of the cervix is systemic, with chemotherapy as the first line of treatment. Surgery and radiotherapy may improve control of local disease but are unlikely significantly to influence the overall prognosis.


Mechanism of homophilic binding mediated by ninjurin, a novel widely expressed adhesion molecule.
Ninjurin is a novel protein that is up-regulated after nerve injury both in dorsal root ganglion (DRG) neurons and in Schwann cells. We previously reported that ninjurin demonstrates properties of a homophilic adhesion molecule and promotes neurite outgrowth from primary cultured DRG neurons. We have now found that ninjurin is widely expressed in both adult and embryonic tissues, primarily in those of epithelial origin. Aggregation assays were used to demonstrate that ninjurin-mediated adhesion requires divalent cations and is an energy-dependent process. The critical domain for ninjurin-mediated homophilic adhesion was localized to an 11-residue region (between Pro26 and Asn37) by mutagenesis and by employing synthetic oligopeptides as competitive inhibitors of ninjurin-mediated adhesion. Of particular importance are the Trp residue at position 29 and the 3 arginines in the region. Furthermore, we show that the peptide which inhibits aggregation of Jurkat cells expressing ninjurin is also capable of blocking the ability of ninjurin to promote neurite extension from DRG neurons. Using FISH analysis, the ninjurin gene was localized to human chromosome 9q22. Several genetic diseases of unknown etiology have been mapped to this region, including hereditary sensory neuropathy type 1, self-healing squamous epithelioma, split-hand/foot deformity type 1, and familial dilated cardiomyopathy.


Electron tomography of cells.
The electron microscope has contributed deep insights into biological structure since its invention nearly 80 years ago. Advances in instrumentation and methodology in recent decades have now enabled electron tomography to become the highest resolution three-dimensional (3D) imaging technique available for unique objects such as cells. Cells can be imaged either plastic-embedded or frozen-hydrated. Then the series of projection images are aligned and back-projected to generate a 3D reconstruction or 'tomogram'. Here, we review how electron tomography has begun to reveal the molecular organization of cells and how the existing and upcoming technologies promise even greater insights into structural cell biology.


Efficiency of energy conversion during shortening of muscle fibres from the dogfish Scyliorhinus canicula.
Force and heat production were measured during isovelocity shortening of tetanized white myotomal muscle fibres from the dogfish at 12 degrees C. For each fibre preparation a range of velocities was used. Mechanical power was calculated from force X velocity of shortening. The rate of total energy output during shortening was evaluated as the sum of mechanical power and the rate of heat production. The ratio of mechanical power to total energy rate was taken as a measure of efficiency of energy conversion to mechanical power during shortening. Efficiency was maximal and varied little in the range of shortening velocities 0.42-0.89 fibres lengths s-1 (0.11-0.23 Vmax); maximal efficiency was 0.33 +/- 0.01 (+/- S.E.M., N = 23 measurements on seven fibre bundles). The efficiency of the white fibres from dogfish was less than that measured in the same way in earlier experiments on frog muscle and tortoise muscle.


Further discussion on the reaction behaviour of triallyl isocyanurate in the UV radiation cross-linking process of polyethylene: a theoretical study.
Further theoretical investigation on the reaction behaviour of triallyl isocyanurate (TAIC) in the UV radiation cross-linking process of polyethylene (PE) is accomplished by density functional theory for high voltage cable insulation materials. The reaction potential energy information of the 13 reaction channels at B3LYP/6-311 + G(d,p) level are identified. These have been explored that the TAIC take part in the reaction behaviour on ground state during UV radiation cross-linking process and TAIC intra-molecular isomerization reaction itself. In addition, the results show that the effect of multiplication and acceleration for the cross-linking reaction of trimethylopropane trimethacrylate (TMPTMA) would be better than that of TAIC. It has further clarified the reasons why UV radiation cross-linking reaction of PE had been initiated by benzophenone (Bp), and the TAIC or TMPTMA needed to take part. The results obtained in the present study could directly guide both the optimization of UV radiation cross-linking PE process and the development of the insulation material of high-voltage cable in real application.


Overview of treatment of vulvovaginal disease.
Vulvovaginal diseases commonly are inadequately diagnosed and treated. Most are dermatologic, but can be atypical in presentation in the warm, moist genital area. There is limited training or education for medical caregivers for these conditions. The first step is correct diagnosis, which requires time and knowledge of the normal anatomy, and careful examination. Dermatologists are invaluable for management as they recognize skin problems and can correct barrier function, control inflammation, and address itching and pain.


2,9-Dicarbonyl-1,10-phenanthroline derivatives with an unprecedented Am(III)/Eu(III) selectivity under highly acidic conditions.
Four lipophilic 1,10-phenanthroline di(thio)amide, diester or diketone derivatives were studied as ligands for Am(III)/Eu(III) separation from acidic media. The synthesis of these compounds is reported together with the extraction tests in different solvents (kerosene, octanol and o-nitrophenyl hexyl ether), HNO3 concentrations and ratios between the ligand and the synergistic agent (Br-Cosan). The promising results obtained from the large number of solvent extraction tests carried out show that it might be possible to apply this class of ligands to advanced reprocessing of spent nuclear fuel. The experimental data indicate that, under the conditions that simulate the real radioactive waste, the extraction efficiency and Am/Eu separation factors are particularly high, thus suggesting that the combination of soft heterocyclic N-donor atoms and hard carbonyl groups of ester and amides affords a tetradentate donor set of atoms (ONNO) that gives rise to remarkable selectivities. ESI-MS studies and DFT calculations shed light on the possible structure of the Eu(3+) complexes indicating that the 1 : 1 : 2 (cation : ligand : anion) complex is slightly more stable than the 1 : 2 : 1 species.


Serosal abrasion of bowel ends does not enhance anastomotic healing.
Anastomotic leakage rates remain unacceptably high, warranting reconsideration of current anastomotic technique. Anastomotic healing may improve by abrading the serosal surface of bowel ends that are invertedly anastomosed, based on the concept that serosal damage evokes inflammatory adherent processes. It is studied if local abrasion leads to stronger anastomoses and reduces leakage. Ninety-eight Wistar rats were allocated to six groups. Either a regular anastomosis (RA) or abraded anastomosis (AA) was constructed in the proximal colon. Animals were sacrificed at day 3 (groups RA3 and AA3, n = 2 × 17) or day 5 (groups RA5 and AA5, n = 2 × 17). Groups RA-Dic and AA-Dic (n = 2 × 15) received diclofenac from day 0 until sacrifice on day 3 to impair anastomotic healing. Outcomes were leakage, bursting pressure, breaking strength, adhesions, and histological appearance. Both in abraded (AA3 and AA5) and control (RA3 and RA5) groups without diclofenac, 1 of 17 anastomoses leaked (6%). Leak rate was 9 of 15 (60%) in group AA-Dic and 8 of 15 (53%) in RA-Dic (P = 1.0). The bursting pressure in group RA3 (127 ± 44 mm Hg) was higher (P = 0.006) compared with group AA3 (82 ± 34 mm Hg), breaking strength was comparable (P = 0.331). Mechanical strength was similar between groups RA5 and AA5. Abrasion did not increase mechanical strength in the diclofenac groups. Adhesion formation was not different between groups. Histology showed dense interserosal scar formation in abraded groups, compared with loose connective tissue in control anastomoses. Abrasion of serosal edges of large bowel ends invertedly anastomosed does not improve anastomotic strength, neither does it reduce leakage in anastomoses compromised by diclofenac.


The Fine-Structure Constant as a Ruler for the Band-Edge Light Absorption Strength of Bulk and Quantum-Confined Semiconductors.
Low-dimensional semiconductors have found numerous applications in optoelectronics. However, a quantitative comparison of the absorption strength of low-dimensional versus bulk semiconductors has remained elusive. Here, we report generality in the band-edge light absorptance of semiconductors, independent of their dimensions. First, we provide atomistic tight-binding calculations that show that the absorptance of semiconductor quantum wells equals mπα (m = 1 or 2 with α as the fine-structure constant), in agreement with reported experimental results. Then, we show experimentally that a monolayer (superlattice) of quantum dots has similar absorptance, suggesting an absorptance quantum of mπα per (confined) exciton diameter. Extending this idea to bulk semiconductors, we experimentally demonstrate that an absorptance quantum equal to mπα per exciton Bohr diameter explains their widely varying absorption coefficients. We thus provided compelling evidence that the absorptance quantum πα per exciton diameter rules the band-edge absorption of all direct semiconductors, regardless of their dimension.


A Prognostic Bio-Model Based on SQSTM1 and N-Stage Identifies Nasopharyngeal Carcinoma Patients at High Risk of Metastasis for Additional Induction Chemotherapy.
Purpose: Metastasis is one of the most important causes of treatment failure in nasopharyngeal carcinoma (NPC). In T4 or N2-3 patients at high-risk of metastasis, concurrent chemoradiotherapy (CCRT) is inadequate and additional induction chemotherapy (IC) is controversial. There is a critical need to develop a better patient stratification to efficiently identify patients at high-risk of metastasis for additional IC. Recently, Sequestosome 1 (SQSTM1)/p62, an autophagy adaptor protein, was identified as one of the metastasis-related proteins in NPC. However, the mechanism by which SQSTM1 is involved in NPC metastasis was not investigated.Experimental Design: The effect of SQSTM1 on cell migration and invasion was examined in vitro and in vivo SQSTM1 expression was analyzed in clinical NPC samples using IHC. Luciferase reporter analyses were conducted to identify the effects of SQSTM1 on NF-κB transcriptional activity. A prediction bio-model was constructed by Cox analysis. Retrospective and prospective randomized clinical data were adopted to build and test the model, respectively.Results: SQSTM1 mediated epithelial to mesenchymal transition (EMT) through the NF-κB pathway to promote NPC metastasis. Inhibiting SQSTM1 enhanced sensitivity to cisplatin in NPC cells. In NPC patients, high SQSTM1 expression was associated with increased risk of distant metastasis. Furthermore, we propose a prognostic bio-model based on SQSTM1 and N-stage to predict NPC metastasis. Most importantly, our prospective randomized study suggested that IC is beneficial for NPC patients with high metastasis risk.Conclusions: The prognostic bio-model identifies NPC patients at high-risk of metastasis for additional IC. Clin Cancer Res; 24(3); 648-58. ©2017 AACR.


Effect of oil droplets and their solid/liquid composition on the phase separation of protein-polysaccharide mixtures.
The phase separation of a model system consisting of sodium caseinate + xanthan ± a low fraction (up to 3 wt %) of an oil-in-water emulsion was studied at room temperature (20-25 °C). The composition of the oil phase was either 100 wt % n-tetradecane (TD); 50% TD + 50% eicosane (EC) or 100% EC. The droplets in these three "emulsions" were therefore totally liquid, partially solid, and totally solid, respectively. In the presence of 22 mM CaCl2, the mixed TD+EC droplets were most effective at inhibiting phase separation, while the EC emulsions could not prevent phase separation at all. At 32 mM CaCl2 the emulsions tended to promote phase separation, possibly due to enhanced calcium ion-induced droplet aggregation. The apparent interfacial viscosity (ηi) between two macroscopically separated phases was also measured. In the presence of the semisolid mixed droplets ηi = 25 mN s m(-1), significantly higher than ηi with the pure (liquid) TD droplets (15 mN s m(-1)) or with the pure solid EC droplets (12 mN s m(-1)) or in the absence of droplets (<3 mN s m(-1)). Confocal microscopy showed that the microstructure of the phase separating regions also depended upon the composition of the oil droplets, and it is tentatively suggested that the more marked effects of the mixed emulsion droplets were due to them forming a stronger network at the interface via partial coalescence. Control of the extent of interfacial aggregation of droplets is therefore possibly one way to influence the course of phase separation in biopolymer mixtures.


Diffusion tensor imaging for spatially-resolved characterization of muscle fiber structure in seafood.
The fiber structure of tissue in meat and seafood has a significant impact on their perceived quality. However, quantifiable description of muscle structure is challenging. We investigate diffusion tensor imaging (DTI) magnetic resonance imaging (MRI) as a method to quantitatively describe tissue structure. DTI measures the anisotropy of water molecule diffusion within muscle fibers. A pilot study evaluated three different cod loin samples: one of high-quality, one of medium-quality, and one of poor-quality. DTI parameters such as fractional anisotropy, axial diffusion and radial diffusion showed clear differences between the sample qualities. Changes in the DTI metrics consistent with freezing and thawing damage to the tissue were observed. The DTI maps were compared to T2-weighted images and DTI detected significant details that were not visible in T2-weighted images. Overall, these results indicate that DTI is a promising method for spatially-resolved characterization of tissue structure in seafood and meat.


Coagulability of arterial and venous blood in healthy subjects, patients with coronary atherosclerosis, and patients with rheumatic cardiopathies.
Coagulability of arterial and venous blood was studied in 50 healthy subjects and in 179 patients with circulation disturbances caused by coronary atherosclerosis and rheumatic cardiopathies. All indicators characterizing the individual haemocoagulation phases were respected. In the healthy subjects the coagulative and fibrinolytic activities of venous blood were higher than those of arterial blood. In the patients with heart failure the coagulability of venous blood decreased and that of arterial blood increased. These phenomena were accompanied by fibrinogenaemia, appearance of fibrinogen B in the blood, elevation of free heparin, and inhibition, but occasional activation, of fibrinolysis in both vascular systems. The findings seem to signalize enhanced intravascular blood coagulability in patients with circulation disturbances, which phenomenon has to be taken into account in the treatment of such patients.


Distinct roles of AKT isoforms in regulating β1-integrin activity, migration, and invasion in prostate cancer.
AKT1 and AKT2 kinases have been shown to play opposite roles in breast cancer migration and invasion. In this study, an RNA interference screen for integrin activity inhibitors identified AKT1 as an inhibitor of β1-integrin activity in prostate cancer. Validation experiments investigating all three AKT isoforms demonstrated that, unlike in breast cancer, both AKT1 and AKT2 function as negative regulators of cell migration and invasion in PC3 prostate cancer cells. Down-regulation of AKT1 and AKT2, but not AKT3, induced activation of cell surface β1-integrins and enhanced adhesion, migration, and invasion. Silencing of AKT1 and AKT2 also resulted in increased focal adhesion size. Importantly, the mechanisms involved in integrin activity regulation were distinct for the two AKT isoforms. Silencing of AKT1 relieved feedback suppression of the expression and activity of several receptor tyrosine kinases, including EGFR and MET, with established cross-talk with β1-integrins. Silencing of AKT2, on the other hand, induced up-regulation of the microRNA-200 (miR-200) family, and overexpression of miR-200 was sufficient to induce integrin activity and cell migration in PC3 cells. Taken together, these data define an inhibitory role for both AKT1 and AKT2 in prostate cancer migration and invasion and highlight the cell type-specific actions of AKT kinases in the regulation of cell motility.


Intravoxel incoherent motion imaging to assess the acute effects of moderate-intensity continuous training and high-intensity interval training on thigh muscles.
This study investigated the use of intravoxel incoherent motion imaging (IVIM) to compare skeletal muscle perfusion during and after high-intensity interval training (HIIT) and moderate-intensity continuous training (MICT) to determine the impact on fat oxidation outcomes. Twenty overweight volunteers were recruited for the study. Each participant received one HIIT intervention and one MICT intervention using a cycling ergometer. Participants underwent a magnetic resonance imaging scan before, immediately after, and 1 and 2 h after each intervention. The IVIM parameters (D, fD*) of the rectus femoris, vastus lateralis, and biceps femoris long head were obtained. Changes in IVIM parameters of these muscles after both exercise interventions were compared using a two-factor repeated measures analysis of variance. In the rectus femoris, the fD* increased immediately after exercise intervention (d = 0.69 × 10-3 mm2 /s, p < 0.0083) and 2 h after exercise intervention (d = 0.64 × 10-3 mm2 /s, p < 0.0083) compared with before exercise. The increase in the fD* in the HIIT group was greater than that in the MICT group (d = 0.32, p = 0.023). In the vastus lateralis, the fD* increased immediately after the exercise intervention (d = 0.53 × 10-3 mm2 /s, p < 0.001) and returned to the pre-exercise level 1 h after exercising. The increase in the fD* in the HIIT group was lower than that in the MICT group (d = -0.21, p = 0.015). For the biceps femoris long head, the fD* was not significantly different between the two exercise interventions before and after exercise. Furthermore, the fD* 60 min after the HIIT intervention correlated with maximal oxygen consumption (VO2max), whereas fD* immediately after the MICT intervention correlated with VO2max. In summary, IVIM parameters can be used to evaluate differences in muscle perfusion between HIIT and MICT, and show a correlation with VO2max.


Taxonomic insights from floral scents of western North American sessile-flowered Trillium.
Chemical composition of floral volatiles can be an important complement to morphological characters in describing and identifying species. Four of the five species of western sessile-flowered Trillium are challenging to distinguish morphologically due to wide intraspecific variation and overlapping characters among taxa. Characterizing their floral volatile compositions could aid future taxonomic, ecological, and evolutionary studies of Trillium and related taxa. We addressed two major questions: How do western sessile Trillium taxa vary in floral chemistry? Can floral scent be used to distinguish species? We collected petals from 600 individuals at 42 wild populations of four sessile Trillium species across California, Oregon, and Washington. Volatile organic compounds from the petals were extracted using solid-phase microextraction, and the volatiles were identified and quantified by gas chromatography-mass spectrometry. The utility of floral scent composition in distinguishing species was tested using nonmetric multidimensional scaling and random forest analysis. Floral volatiles of the white-petaled T. albidum were dominated by oxygenated monoterpenes and showed considerable geographic variation that paralleled morphological variation. The maroon-petaled T. angustipetalum and T. kurabayashii produced floral scents characterized by aliphatic esters, but each had a distinct chemical composition. Petal color of Trillium chloropetalum is highly variable, as were its scent compositions, which were blends of volatiles from both white-petaled and maroon-petaled congeneric taxa. Differences in floral scent compositions are consistent with current taxonomy of the western sessile Trillium group. In cases where species delimitations are difficult based on morphology, floral scent composition provides taxonomic insight and suggests a potential hybrid origin for T. chloropetalum.


Role of Rhodobacter sphaeroides photosynthetic reaction center residue M214 in the composition, absorbance properties, and conformations of H(A) and B(A) cofactors.
In the native reaction center (RC) of Rhodobacter sphaeroides, the side chain of (M)L214 projects orthogonally toward the plane and into the center of the A branch bacteriopheophytin (BPhe) macrocycle. The possibility that this side chain is responsible for the dechelation of the central Mg(2+) of bacteriochlorophyll (BChl) was investigated by replacement of (M)214 with residues possessing small, nonpolar side chains that can neither coordinate nor block access to the central metal ion. The (M)L214 side chain was also replaced with Cys, Gln, and Asn to evaluate further the requirements for assembly of the RC with BChl in the HA pocket. Photoheterotrophic growth studies showed no difference in growth rates of the (M)214 nonpolar mutants at a low light intensity, but the growth of the amide-containing mutants was impaired. The absorbance spectra of purified RCs indicated that although absorbance changes are associated with the nonpolar mutations, the nonpolar mutant RC pigment compositions are the same as in the wild-type protein. Crystal structures of the (M)L214G, (M)L214A, and (M)L214N mutants were determined (determined to 2.2-2.85 Å resolution), confirming the presence of BPhe in the HA pocket and revealing alternative conformations of the phytyl tail of the accessory BChl in the BA site of these nonpolar mutants. Our results demonstrate that (i) BChl is converted to BPhe in a manner independent of the aliphatic side chain length of nonpolar residues replacing (M)214, (ii) BChl replaces BPhe if residue (M)214 has an amide-bearing side chain, (iii) (M)214 side chains containing sulfur are not sufficient to bind BChl in the HA pocket, and (iv) the (M)214 side chain influences the conformation of the phytyl tail of the BA BChl.


Effect of smartphone applications on cardiopulmonary resuscitation quality metrics in a mannequin study: A randomized trial.
The aim of this randomized, cross-over trial is to reveal the effect of smartphone cardio-pulmonary resuscitation (CPR) feedback applications (App) on a group of lay rescuers' chest compression-only CPR quality metrics. Quality metrics is measured initially and after 3 months. A floor-based Resusci Anne mannequin (Laerdal Medical, Stavanger, Norway) was used. Three scenarios (CPR with device App-on [scenario-a], CPR with device App-off [scenario-b], and hands-only CPR [scenario-c]) were randomly allocated to all participants. All the participants performed 2 min of hands only-CPR for each scenario. Data of mean chest compression rate, mean chest compression depth, and recoil were recorded and compared for each scenario. One hundred and thirty-seven first-year students from the Vocational School of Health Services in Turkey participated in this study to mimic lay rescuers. Difference in the initial mean rate of chest compressions was statistically significant when CPR was performed with device App-on (scenario-a) compared to scenarios b and c (P < 0.001, P < 0.001). Furthermore, difference in the mean chest compression rate at the 3rd month was statistically significant among the scenarios when CPR was performed with device App-on (scenario-a) (P = 0.002, P = 0.001). The difference in initial and 3rd month mean compression depth and the percentage of recoil was not statistically significant among the scenarios. This study shows that the mean chest compression rate and percentage of compressions with adequate rate improved with smartphone App-on, and these results were persistent up to 3 months.


Optimizing edges and defects of supported MoS2 catalysts for hydrogen evolution via an external electric field.
As a fascinating non-precious catalyst for hydrogen evolution reaction (HER), two-dimensional (2D) molybdenum disulphide (MoS2) has attracted ever-growing interest. While the pristine basal plane of MoS2 is chemically inactive, certain edges and defects have been recognized to be catalytically active for HER. Nevertheless, the per-site activity of MoS2 is still much lower than that of Pt. Therefore, further optimization of active sites becomes highly desirable to enhance the overall catalytic activity of MoS2. In this work, we propose to use an electric field to engineer the electronic structure of edges and defects of MoS2, aiming to optimize its catalytic performance. Via systematic density functional theory based first-principles calculations, we investigated the adsorption of H atoms on different edges of free-standing and supported MoS2, revealing the critical role of S p-resonance states near the Fermi level in determining H adsorption, which offers an excellent descriptor for the catalytic activity associated with the electronic structure. Remarkably, by introducing an external electric field, we demonstrate the ability to fine tune the position of S p-resonance states, which can give an optimal H adsorption strength on MoS2 for HER. We also explored field effects on S vacancies in the basal plane, which show a different behavior for H adsorption due to the presence of Mo d states that are insensitive to the electric field. We expect these findings to shed new light on the design and control of MoS2-based catalysts for industrial applications.


Astrocyte Subtype Vulnerability in Stem Cell Models of Vanishing White Matter.
Astrocytes have gained attention as important players in neurological disease. In line with their heterogeneous character, defects in specific astrocyte subtypes have been identified. Leukodystrophy vanishing white matter (VWM) shows selective vulnerability in white matter astrocytes, but the underlying mechanisms remain unclear. Induced pluripotent stem cell technology is being extensively explored in studies of pathophysiology and regenerative medicine. However, models for distinct astrocyte subtypes for VWM are lacking, thereby hampering identification of disease-specific pathways. Here, we characterize human and mouse pluripotent stem cell-derived gray and white matter astrocyte subtypes to generate an in vitro VWM model. We examined morphology and functionality, and used coculture methods, high-content microscopy, and RNA sequencing to study VWM cultures. We found intrinsic vulnerability in specific astrocyte subpopulations in VWM. When comparing VWM and control cultures, white matter-like astrocytes inhibited oligodendrocyte maturation, and showed affected pathways in both human and mouse cultures, involving the immune system and extracellular matrix. Interestingly, human white matter-like astrocytes presented additional, human-specific disease mechanisms, such as neuronal and mitochondrial functioning. Astrocyte subtype cultures revealed disease-specific pathways in VWM. Cross-validation of human- and mouse-derived protocols identified human-specific disease aspects. This study provides new insights into VWM disease mechanisms, which helps the development of in vivo regenerative applications, and we further present strategies to study astrocyte subtype vulnerability in neurological disease. ANN NEUROL 2019;86:780-792.


The novel aerodynamics of insect flight: applications to micro-air vehicles.
The wing motion in free flight has been described for insects ranging from 1 to 100 mm in wingspan. To support the body weight, the wings typically produce 2-3 times more lift than can be accounted for by conventional aerodynamics. Some insects use the fling mechanism: the wings are clapped together and then flung open before the start of the downstroke, creating a lift-enhancing vortex around each wing. Most insects, however, rely on a leading-edge vortex (LEV) created by dynamic stall during flapping; a strong spanwise flow is also generated by the pressure gradients on the flapping wing, causing the LEV to spiral out to the wingtip. Technical applications of the fling are limited by the mechanical damage that accompanies repeated clapping of the wings, but the spiral LEV can be used to augment the lift production of propellers, rotors and micro-air vehicles (MAVs). Design characteristics of insect-based flying machines are presented, along with estimates of the mass supported, the mechanical power requirement and maximum flight speeds over a wide range of sizes and frequencies. To support a given mass, larger machines need less power, but smaller ones operating at higher frequencies will reach faster speeds.


Seizure outcome and use of antiepileptic drugs after epilepsy surgery according to histopathological diagnosis: a retrospective multicentre cohort study.
Surgery is a widely accepted treatment option for drug-resistant focal epilepsy. A detailed analysis of longitudinal postoperative seizure outcomes and use of antiepileptic drugs for different brain lesions causing epilepsy is not available. We aimed to analyse the association between histopathology and seizure outcome and drug freedom up to 5 years after epilepsy surgery, to improve presurgical decision making and counselling. In this retrospective, multicentre, longitudinal, cohort study, patients who had epilepsy surgery between Jan 1, 2000, and Dec 31, 2012, at 37 collaborating tertiary referral centres across 18 European countries of the European Epilepsy Brain Bank consortium were assessed. We included patients of all ages with histopathology available after epilepsy surgery. Histopathological diagnoses and a minimal dataset of clinical variables were collected from existing local databases and patient records. The primary outcomes were freedom from disabling seizures (Engel class 1) and drug freedom at 1, 2, and 5 years after surgery. Proportions of individuals who were Engel class 1 and drug-free were reported for the 11 main categories of histopathological diagnosis. We analysed the association between histopathology, duration of epilepsy, and age at surgery, and the primary outcomes using random effects multivariable logistic regression to control for confounding. 9147 patients were included, of whom seizure outcomes were available for 8191 (89·5%) participants at 2 years, and for 5577 (61·0%) at 5 years. The diagnoses of low-grade epilepsy associated neuroepithelial tumour (LEAT), vascular malformation, and hippocampal sclerosis had the best seizure outcome at 2 years after surgery, with 77·5% (1027 of 1325) of patients free from disabling seizures for LEAT, 74·0% (328 of 443) for vascular malformation, and 71·5% (2108 of 2948) for hippocampal sclerosis. The worst seizure outcomes at 2 years were seen for patients with focal cortical dysplasia type I or mild malformation of cortical development (50·0%, 213 of 426 free from disabling seizures), those with malformation of cortical development-other (52·3%, 212 of 405 free from disabling seizures), and for those with no histopathological lesion (53·5%, 396 of 740 free from disabling seizures). The proportion of patients being both Engel class 1 and drug-free was 0-14% at 1 year and increased to 14-51% at 5 years. Children were more often drug-free; temporal lobe surgeries had the best seizure outcomes; and a longer duration of epilepsy was associated with reduced chance of favourable seizure outcomes and drug freedom. This effect of duration was evident for all lesions, except for hippocampal sclerosis. Histopathological diagnosis, age at surgery, and duration of epilepsy are important prognostic factors for outcomes of epilepsy surgery. In every patient with refractory focal epilepsy presumed to be lesional, evaluation for surgery should be considered. None.


Zoledronic acid initiated during the first year of androgen deprivation therapy increases bone mineral density in patients with prostate cancer.
Androgen deprivation therapy in patients with prostate cancer is associated with bone loss and an increased risk of fractures. Zoledronic acid protects against bone mineral density loss when initiated concurrently with androgen deprivation therapy. We evaluated the effect of zoledronic acid initiated subsequent to androgen deprivation therapy on bone mineral density and biochemical markers of bone turnover. Patients with prostate cancer without bone metastases who had received androgen deprivation therapy for 12 months or less were randomized to 4 mg zoledronic acid or placebo intravenously every 3 months for 1 year. Patients were stratified according to androgen deprivation therapy duration (less than 6 vs 6 to 12 months). The primary end point was the change in femoral neck and lumbar spine bone mineral density in the 2 groups. The secondary end point was the change in serum bone specific alkaline phosphatase and urine N-telopeptide levels. Total hip bone mineral density was also measured. The 120 patients with prostate cancer received zoledronic acid (61) or placebo (59). Compared with placebo, zoledronic acid increased femoral neck, total hip and lumbar spine bone mineral density yearly by 3.6% (p = 0.0004), 3.8% (p <0.0001) and 6.7% (p <0.0001), respectively. The effects of zoledronic acid on bone mineral density at these 3 sites were not differentiated according to androgen deprivation therapy duration. Additionally, mean bone specific alkaline phosphatase and N-telopeptide levels were decreased in the zoledronic acid group (each p <0.0001) and were increased in the placebo group (p <0.0001 and p = 0.004, respectively). Zoledronic acid increased bone mineral density and suppressed bone turnover markers in patients with prostate cancer without bone metastases when initiated during year 1 of androgen deprivation therapy.


Sub-threshold autism traits: the role of trait emotional intelligence and cognitive flexibility.
Theory and research suggests that features of autism are not restricted to individuals diagnosed with autism spectrum disorders (ASDs), and that autism-like traits vary throughout the general population at lower severities. The present research first investigated the relationship of autism traits with trait emotional intelligence and empathy in a sample of 163 adults aged between 18 and 51 years (44% male). It then examined performance on a set of tasks assessing social cognition and cognitive flexibility in 69 participants with either high or low scores on ASD traits. Results confirm that there is pronounced variation within the general population relating to ASD traits, which reflect similar (though less severe) social-cognitive and emotional features to those observed in ASDs.


Population pharmacokinetics of ceftaroline in patients with acute bacterial skin and skin structure infections or community-acquired bacterial pneumonia.
Ceftaroline, the active form of ceftaroline fosamil, is a broad-spectrum cephalosporin antibiotic. A population pharmacokinetic (PPK) model for ceftaroline was developed in NONMEM® using data from 185 healthy subjects and 92 patients with acute bacterial skin and skin structure infection (ABSSSI). Data from 128 patients with community-acquired bacterial pneumonia (CABP) were used for external model validation. Healthy subjects received 50-2,000 mg ceftaroline fosamil via intravenous (IV) infusion over 1 hour or intramuscular (IM) injection q12h or q24h. ABSSSI and CABP patients received 600 mg of ceftaroline fosamil IV over 1 hour q12h. A three-compartment model with zero-order IV or parallel first-order IM input and first-order elimination described ceftaroline fosamil PK. A two-compartment model with first-order conversion of prodrug to ceftaroline and parallel linear and saturable elimination described ceftaroline PK. Creatinine clearance was the primary determinant of ceftaroline exposure. Good agreement between the observed data and both population (r(2) = 0.93) and individual post-hoc (r(2) = 0.98) predictions suggests the PPK model can adequately approximate ceftaroline PK using covariate information. Such a PPK model can evaluate dose adjustments for patients with renal impairment and generate ceftaroline exposures for use in pharmacokinetic-pharmacodynamic assessments of efficacy in patients with ABSSSI or CABP.


Plasma levels of tumor necrosis factor-alpha, interferon-gamma, inducible nitric oxide synthase, and 3-nitrotyrosine in drug-resistant and drug-sensitive pulmonary tuberculosis patients, Ibadan, Nigeria.
Nigeria is one of the countries with a high burden of tuberculosis (TB) in the world. TB associated inflammation is reported to be central to progression from latent TB to active TB or drug sensitive TB (DSTB) to drug resistant TB (DRTB). Inflammatory cytokines, especially interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α), act synergistically in the control of TB infection. They activate macrophages to produce effector molecules such as inducible nitric oxide synthase (iNOS), nitric oxide, and ultimately 3-nitrotyrosines(3-NTs), which are involved in the control of TB. This study investigated the potential involvement of TNF-α, IFN-γ, iNOS, and 3-NT in differentiating DRTB and DSTB in Ibadan, Nigeria. One hundred participants above 18 years were recruited into this study and were grouped as follows: 32 DRTB, 34 DSTB, and 34 apparently healthy controls. Plasma from the patients was used for the analyses of inflammatory (TNF α and IFN-γ) and oxidative stress (iNOS and 3-NT) biomarkers using the ELISA. Mann-Whitney test was applied for the statistical test. Mean levels of plasma TNF-α, IFN-γ, iNOS, and 3-NT were higher in DRTB (19.74 ± 3.62 pg/mL, 4.41 ± 0.96 pg/mL, 1791.07 ± 419.42 pg/mL, and 20.27 ± 1.80 ng/mL, respectively) and DSTB (17.02 ± 1.84 pg/mL, 5.59 ± 1.40 pg/mL, 2823.42 ± 685.32 pg/mL, and 25.06 ± 2.15 ng/mL, respectively) compared with controls (12.18 ± 0.92 pg/mL, 1.58 ± 0.21 pg/mL, 1275.86 ± 166.12 pg/mL, and 19.98 ± 1.23 ng/mL, respectively). In addition, higher plasma levels of IFN-γ (P > 0.05), iNOS (P > 0.05), and 3-NT (P < 0.05) were observed in DSTB compared with DRTB patients. The 3-NT may be used as differentiating markers of DSTB from DRTB.


Embryos of non-native anoles are robust to urban thermal environments.
The transformation of natural habitats into urban landscapes dramatically alters thermal environments, which in turn, can impact local biota. Ectothermic organisms that are oviparous are particularly sensitive to these altered environments because their embryos cannot behaviorally thermoregulate and the surrounding environment determines the temperature experienced during development. We studied the effects of urban and forested thermal environments on embryo development and hatchling phenotypes in two non-native lizards (Anolis sagrei and A. cristatellus) in metropolitan Miami, Florida. To determine if embryos from urban and forested sites are adapted to their respective thermal environments, we incubated eggs from each site using temperatures that simulate likely nest conditions in both urban and forested environments. For both species, urban thermal environments accelerated embryonic development, but had no impact on egg survival or any of the phenotypic traits that were measured (e.g., body size, running performance, and locomotor behavior). Our results provide no evidence that embryos from urban and forested sites are adapted to their respective thermal environments. Instead, the lack of any major effects suggest that embryos of both species are physiologically robust with respect to novel environments, which could have facilitated their success in establishing in non-native ranges and in human-modified landscapes.


Intracellular Hg(0) Oxidation in Desulfovibrio desulfuricans ND132.
The disposal of elemental mercury (Hg(0)) wastes in mining and manufacturing areas has caused serious soil and groundwater contamination issues. Under anoxic conditions, certain anaerobic bacteria can oxidize dissolved elemental mercury and convert the oxidized Hg to neurotoxic methylmercury. In this study, we conducted experiments with the Hg-methylating bacterium Desulfovibrio desulfuricans ND132 to elucidate the role of cellular thiols in anaerobic Hg(0) oxidation. The concentrations of cell-surface and intracellular thiols were measured, and specific fractions of D. desulfuricans ND132 were examined for Hg(0) oxidation activity and analyzed with extended X-ray absorption fine structure (EXAFS) spectroscopy. The experimental data indicate that intracellular thiol concentrations are approximately six times higher than those of the cell wall. Cells reacted with a thiol-blocking reagent were severely impaired in Hg(0) oxidation activity. Spheroplasts lacking cell walls rapidly oxidized Hg(0) to Hg(II), while cell wall fragments exhibited low reactivity toward Hg(0). EXAFS analysis of spheroplast samples revealed that multiple different forms of Hg-thiols are produced by the Hg(0) oxidation reaction and that the local coordination environment of the oxidized Hg changes with reaction time. The results of this study indicate that Hg(0) oxidation in D. desulfuricans ND132 is an intracellular process that occurs by reaction with thiol-containing molecules.


Metabolic profile and hepatoprotective effect of Aeschynomene elaphroxylon (Guill. & Perr.).
Liver diseases are life-threatening and need urgent medical treatments. Conventional treatment is expensive and toxic, so the urge for nutraceutical hepatoprotective agents is crucial. This study is considered the first metabolic profile of Aeschynomene elaphroxylon (Guill. & Perr.) extracts of; flowers, leaves & bark adopting UPLC-Orbitrap HRMS analysis to determine their bioactive metabolites, and it was designed to investigate the potential hepatoprotective activity of A. elaphroxylon flowers and bark extracts against CCl4-induced hepatic fibrosis in rats. Forty-nine compounds of various classes were detected in the three extracts, with triterpenoid saponins as the major detected metabolite. Flowers and bark extracts presented similar chemical profile while leaves extract was quite different. The antioxidant activities of the flowers, leaves & bark extracts were measured by in vitro assays as Fe+3 reducing antioxidant power and Oxygen radical absorbance capacity. It revealed that flowers and bark extracts had relatively high antioxidant activity as compared to leaves extract. Based on the metabolic profile and in vitro antioxidant activity, flowers and bark ethanolic extracts were chosen for alleviation of hepatotoxicity induced by CCl4 in rats. The hepatoprotective activity was studied through measuring hepatotoxicity biomarkers in serum (ALT, AST, and Albumin). Liver tissues were examined histopathologically and their homogenates were used in determining the intracellular levels of oxidative stress biomarkers (MDA, GSH), inflammatory markers (TNF-α). Flowers and bark ethanolic extracts exerted a significant hepatoprotective effect through reduction in the activities of ALT, AST and Albumin, the tested extracts reduced oxidative stress by increasing GSH content and reducing the MDA level. Furthermore, the extracts decreased levels of pro-inflammatory TNF-α. Moreover, the present study revealed the potentiality of A. elaphroxylon in ameliorating the CCl4-induced hepatic fibrosis in rats. In this aspect, A. elaphroxylon can be used with other agents as a complementary drug.


Antibacterial Surfaces with Activity against Antimicrobial Resistant Bacterial Pathogens and Endospores.
Hospital-acquired bacterial infections are a significant burden on healthcare systems worldwide causing an increased duration of hospital stays and prolonged patient suffering. We show that polyurethane containing crystal violet (CV) and 3-4 nm zinc oxide nanoparticles (ZnO NPs) possesses excellent bactericidal activity against hospital-acquired pathogens including multidrug resistant Escherichia coli (E. coli), Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus (MRSA), and even highly resistant endospores of Clostridioides (Clostridium) difficile. Importantly, we used clinical isolates of bacterial strains, a protocol to mimic the environmental conditions of a real exposure in the healthcare setting, and low light intensity equivalent to that encountered in UK hospitals (∼500 lux). Our data shows that ZnO NPs enhance the photobactericidal activity of CV under low intensity light even with short exposure times, and we show that this involves both Type I and Type II photochemical pathways. Interestingly, polyurethane containing ZnO NPs alone showed significant bactericidal activity in the dark against one strain of E. coli, indicating that the NPs possess both light-activated synergistic activity with CV and inherent bactericidal activity that is independent of light. These new antibacterial polymers are potentially useful in healthcare facilties to reduce the transmission of pathogens between people and the environment.


Inducible glutathione S-transferase (IrGST1) from the tick Ixodes ricinus is a haem-binding protein.
Blood-feeding parasites are inadvertently exposed to high doses of potentially cytotoxic haem liberated upon host blood digestion. Detoxification of free haem is a special challenge for ticks, which digest haemoglobin intracellularly. Ticks lack a haem catabolic mechanism, mediated by haem oxygenase, and need to dispose of vast majority of acquired haem via its accumulation in haemosomes. The knowledge of individual molecules involved in the maintenance of haem homeostasis in ticks is still rather limited. RNA-seq analyses of the Ixodes ricinus midguts from blood- and serum-fed females identified an abundant transcript of glutathione S-transferase (gst) to be substantially up-regulated in the presence of red blood cells in the diet. Here, we have determined the full sequence of this encoding gene, ir-gst1, and found that it is homologous to the delta-/epsilon-class of GSTs. Phylogenetic analyses across related chelicerates revealed that only one clear IrGST1 orthologue could be found in each available transcriptome from hard and soft ticks. These orthologues create a well-supported clade clearly separated from other ticks' or mites' delta-/epsilon-class GSTs and most likely evolved as an adaptation to tick blood-feeding life style. We have confirmed that IrGST1 expression is induced by dietary haem(oglobin), and not by iron or other components of host blood. Kinetic properties of recombinant IrGST1 were evaluated by model and natural GST substrates. The enzyme was also shown to bind haemin in vitro as evidenced by inhibition assay, VIS spectrophotometry, gel filtration, and affinity chromatography. In the native state, IrGST1 forms a dimer which further polymerises upon binding of excessive amount of haemin molecules. Due to susceptibility of ticks to haem as a signalling molecule, we speculate that the expression of IrGST1 in tick midgut functions as intracellular buffer of labile haem pool to ameliorate its cytotoxic effects upon haemoglobin intracellular hydrolysis.


2,7-Dimethoxynaphthalene.
C12H12O2, Mr = 188.2, orthorhombic, P2(1)2(1)2(1), a = 6.109 (3), b = 8.235 (2), c = 19.713 (3) A, V = 991.8 (9) A3, Z = 4, Dx = 1.260 g cm-3, lambda(Mo K alpha) = 0.71073 A, mu = 0.79 cm-1, F(000) = 400, T = 293 K, R = 0.047 for 737 observations (of 1337 unique data). The average deviation from planarity is 0.013 (3) A with a maximum of 0.027 (3) A for the fused rings. Steric interaction between the methyl groups and the adjacent H atoms is responsible for opening the angles between the methoxyl groups and the rings. The CH3O-C-C angles are 125.5 (3) degrees [125.0 (3) degrees] where the methyl group is syn to the hydrogen, and 114.3 (3) degrees [114.2 (3) degrees] where it is anti.


Imprinted genes and the environment: links to the toxic metals arsenic, cadmium, lead and mercury.
Imprinted genes defy rules of Mendelian genetics with their expression tied to the parent from whom each allele was inherited. They are known to play a role in various diseases/disorders including fetal growth disruption, lower birth weight, obesity, and cancer. There is increasing interest in understanding their influence on environmentally-induced disease. The environment can be thought of broadly as including chemicals present in air, water and soil, as well as food. According to the Agency for Toxic Substances and Disease Registry (ATSDR), some of the highest ranking environmental chemicals of concern include metals/metalloids such as arsenic, cadmium, lead and mercury. The complex relationships between toxic metal exposure, imprinted gene regulation/expression and health outcomes are understudied. Herein we examine trends in imprinted gene biology, including an assessment of the imprinted genes and their known functional roles in the cell, particularly as they relate to toxic metals exposure and disease. The data highlight that many of the imprinted genes have known associations to developmental diseases and are enriched for their role in the TP53 and AhR pathways. Assessment of the promoter regions of the imprinted genes resulted in the identification of an enrichment of binding sites for two transcription factor families, namely the zinc finger family II and PLAG transcription factors. Taken together these data contribute insight into the complex relationships between toxic metals in the environment and imprinted gene biology.


Chronological aging impacts abundance, function and microRNA content of extracellular vesicles produced by human epidermal keratinocytes.
The disturbance of intercellular communication is one of the hallmarks of aging. The goal of this study is to clarify the impact of chronological aging on extracellular vesicles (EVs), a key mode of communication in mammalian tissues. We focused on epidermal keratinocytes, the main cells of the outer protective layer of the skin which is strongly impaired in the skin of elderly. EVs were purified from conditioned medium of primary keratinocytes isolated from infant or aged adult skin. A significant increase of the relative number of EVs released from aged keratinocytes was observed whereas their size distribution was not modified. By small RNA sequencing, we described a specific microRNA (miRNA) signature of aged EVs with an increase abundance of miR-30a, a key regulator of barrier function in human epidermis. EVs from aged keratinocytes were found to be able to reduce the proliferation of young keratinocytes, to impact their organogenesis properties in a reconstructed epidermis model and to slow down the early steps of skin wound healing in mice, three features observed in aged epidermis. This work reveals that intercellular communication mediated by EVs is modulated during aging process in keratinocytes and might be involved in the functional defects observed in aged skin.


Postoperative Clostridium difficile-associated diarrhea.
Abdominal surgery is thought to be a risk factor for Clostridium difficile-associated diarrhea (CDAD). The aims of this study were to discern pre-operative factors associated with postoperative CDAD, examine outcomes after postoperative CDAD, and compare outcomes of postoperative versus medical CDAD. Data from 3904 patients who had abdominal operations at Montefiore Medical Center were extracted from Montefiore's clinical information system. Cases of 30-day postoperative CDAD were identified. Pre-operative factors associated with developing postoperative CDAD were identified using logistic regression. Medical patients and surgical patients with postoperative CDAD were compared for demographic and clinical characteristics, CDAD recurrence, and 90-day postinfection mortality. The rate of 30-day postoperative CDAD was 1.2%. After adjustment for age and comorbidities, factors significantly associated with postoperative CDAD were: antibiotic use (OR: 1.94), proton pump inhibitor (PPI) use (OR: 2.32), prior hospitalization (OR: 2.27), and low serum albumin (OR: 2.05). In comparison with medical patients with CDAD, postoperative patients with CDAD were significantly more likely to have received antibiotics (98% vs 85%), less likely to have received a PPI (39% vs 58%), or to have had a prior hospitalization (43% vs 67%). Postoperative patients with CDAD had decreased risk of mortality when compared with medical patients with CDAD (HR 0.36). CDAD is an infrequent complication after abdominal operations. Several avoidable pre-operative exposures (eg, antibiotic and PPI use) were identified that increase the risk of postoperative CDAD. Postoperative CDAD is associated with decreased risk of mortality when compared with CDAD on the medical service.


Plasma and platelet von Willebrand factor abnormalities in patients with uremia: lack of correlation with uremic bleeding.
Chronic renal failure often is associated with abnormal bleeding that may represent an important complication of this disorder. The hemorrhagic tendency currently is attributed to altered primary hemostasis, mainly platelet dysfunction. However, von Willebrand factor (vWF) also seems to be involved, even though the nature of its abnormalities is still controversial. To gain insight into the role of vWF in determining uremic bleeding, we studied 11 patients with stable, chronic renal failure. We found a significant increase in plasma factor VIII (FVIII), vWF:antigen (Ag), and vWF:ristocetin cofactor (Rco) levels, associated with a mean decrease in platelet vWF:Ag. Plasma vWF multimer pattern was characterized by increased representation of all oligomers in all patients, but five patients also showed a slight decrease in large vWF multimers. In addition, platelet vWF multimer pattern displayed a decrease in all components, especially those with high molecular weight. Despite normal bleeding time, collagen-induced platelet aggregation was defective in almost all patients, whereas vWF collagen binding capacity was normal. The levels of glycocalicin, the circulating fragment of glycoprotein Ib-IX, the major platelet vWF receptor, were also normal. In six patients who also were studied after initiation of dialysis, collagen-induced platelet aggregation was impaired further. Moreover, plasma vWF, and especially FVIII levels, were increased additionally, in association with a normalized platelet vWF content and an improved vWF multimer pattern. The results suggest that vWF abnormalities are present in uremia. Moreover, thrombopathy caused by impaired collagen-induced platelet aggregation is constantly present and apparently not improved by dialytic treatment.


A peptide-modified solid lipid nanoparticle formulation of paclitaxel modulates immunity and outperforms dacarbazine in a murine melanoma model.
Melanoma is a highly aggressive skin cancer. A paclitaxel formulation of solid lipid nanoparticles modified with Tyr-3-octreotide (PSM) is employed to treat melanoma that highly expresses somatostatin receptors (SSTRs). PSM exerts more apoptotic and anti-invasive effects in B16F10 mice melanoma cells as compared to dacarbazine (DTIC), an approved chemotherapeutic drug for treating aggressive melanoma. Besides, PSM induces one of the biomarkers of immunogenic cell death in vitro and in vivo as confirmed by calreticulin exposure on the B16F10 cell surface. We observed a significant number of CD8 positive T cells in the tumor bed of the PSM treated group. As a result, PSM effectively reduces tumor volume in vivo as compared to DTIC. PSM also induces a favorable systemic immune response as determined in the spleen and sera of the treated animals. Importantly, PSM can reduce the number of nodule formations in the experimental lung metastasis model. Our experimentations indicate that the metronomic PSM exhibits remarkable anti-melanoma activities without any observable toxicity. This immune modulation behavior of PSM can be exploited for the therapy of melanoma and probably for other malignancies.


Canine notochordal cell-secreted factors protect murine and human nucleus pulposus cells from apoptosis by inhibition of activated caspase-9 and caspase-3/7.
Introduction Effective therapies that may stop or even reverse disc degeneration remain elusive. A minimally invasive method through which nucleus pulposus (NP) cell viability could be achieved would revolutionize the treatment of degenerative disc disease (DDD). With the presented work, we have investigated if nonchondrodystrophic (NCD) canine intervertebral disc (IVD)-derived notochordal cell conditioned medium (NCCM) and chondrodystrophic (CD) canine IVD-derived conditioned medium (CDCM) are able to protect murine and human NP cells from apoptosis. Materials and Methods We developed NCCM and CDCM from hypoxic culture of freshly isolated NPs from NCD and CD canines, respectively. We obtained murine NP cells from nine different C57BL/6 mice and human NP cells from four patients who underwent surgery for discectomy. The cells were cultured with ADMEM/F-12 (control media), NCCM, or CDCM under hypoxic conditions (3.5% O2) and treated with IL-1β + FasL or Etoposide. All media were supplemented with 2% fetal bovine serum. We then determined the expression of specific apoptotic pathways in the murine and human NP cells by recording activated caspase-8, caspase-9, and caspase-3/7 activity. Results In the murine NP cells, NCCM inhibits IL-1β + FasL- and Etoposide-mediated apoptosis via suppression of activated caspase-9 and caspase-3/7, CDCM demonstrated an inhibitory effect on IL-1β + FasL-mediated apoptosis via caspase-3/7 (Fig. 1A). In the human NP cells, NCCM inhibits Etoposide- mediated apoptosis via suppression of activated caspase-8, caspase-9, and mainly caspase-3/7. CDCM demonstrated an inhibitory effect on Etoposide-mediated apoptosis via suppression of activated caspase-8, caspase-9, and mainly caspase-3/7, though not as effective as NCCM (Fig. 1B). Conclusion IL-1β + FasL are known key molecules in the progression of DDD. Here, we demonstrate that soluble factors secreted by the NCD IVD NP strongly protect murine NP cells not only from IL-1β + FasL but also from Etoposide-induced apoptosis via suppression of activated caspase-9 and caspase-3/7. In the human samples, addition of IL-1β + FasL did not increase cell death. Because the human cell samples were obtained from herniated discs that are probably already undergoing a degenerative process, it is likely that there was already some degree of activation by the endogenously secreted prodegenerative factors such as IL-1β + FasL. It may be that the NP cells, once they have reached a pivotal point of the degenerative cascade, no longer respond to exogenously applied IL-1β + FasL in contrast to the otherwise "healthy" discs obtained from the mice. Interestingly, the rescue effect of NCCM in the etoposide-treated cells (murine and human) suggests that NCCM is capable of influencing the signaling pathways known to be relevant to etoposide-induced cell death. A better understanding and harnessing of the restorative powers of the notochordal cell could lead to novel cellular and molecular strategies for the treatment of DDD.


Ridge-based vessel segmentation in color images of the retina.
A method is presented for automated segmentation of vessels in two-dimensional color images of the retina. This method can be used in computer analyses of retinal images, e.g., in automated screening for diabetic retinopathy. The system is based on extraction of image ridges, which coincide approximately with vessel centerlines. The ridges are used to compose primitives in the form of line elements. With the line elements an image is partitioned into patches by assigning each image pixel to the closest line element. Every line element constitutes a local coordinate frame for its corresponding patch. For every pixel, feature vectors are computed that make use of properties of the patches and the line elements. The feature vectors are classified using a kappaNN-classifier and sequential forward feature selection. The algorithm was tested on a database consisting of 40 manually labeled images. The method achieves an area under the receiver operating characteristic curve of 0.952. The method is compared with two recently published rule-based methods of Hoover et al. and Jiang et al. The results show that our method is significantly better than the two rule-based methods (p < 0.01). The accuracy of our method is 0.944 versus 0.947 for a second observer.


Phased-ultrasonic receiving-planar array transducer for partial discharge location in transformer.
Partial discharge (PD) location in transformers is very important, and many methods that have been brought forward in past decades have a limitation theoretically, namely, they cannot locate multiple PDs in electrical equipment. In this paper, a new PD location method based on UHF and ultrasonic-phased arrays receiving theory has been presented, which has a possibility to locate multiple PDs. According to the method, a phased-ultrasonic receiving-planar array sensor that possesses 16 * 16 elements is designed; and, based on the phased-array theory, the characteristics of the plane sensor are studied. The laboratory experimental tests on the plane sensor element indicates that it has a good performance within the frequency band of the main ultrasonic energy produced by PD in transformer oil. Location tests are conducted on one or two piezoelectric ultrasonic sources in oil, which are both simulated as PD sources and triggered by an electrical pulse whose front is considered as a time benchmark in the locating algorithm. The test results show locations to one and two PDs can be realized in a single measurement, which lays a foundation for locating PDs in a power transformer in service.


Kuala Lumpur train collision during the COVID-19 pandemic.
Two city trains collided in an underground tunnel on 24 May 2021 at the height of COVID-19 pandemic near the Petronas Towers, Kuala Lumpur, Malaysia, immediately after the evening rush hours. We aim to evaluate the management of this mass casualty incident highlighting the lessons learned to be used in preparedness for similar incidents that may occur in other major cities worldwide. Information regarding incident site and hospital management response were analysed. Data on demography, triaging, injuries and hospital management of patients were collected according to a designed protocol. Challenges, difficulties and their solutions were reported. The train's emergency response team (ERT) has shut down train movements towards the incident site. Red zone (in the tunnel), yellow zone (the station platform) and green zone (outside the station entrance) were established. The fire and rescue team arrived and assisted the ERT in the red zone. Incident command system was established at the site. Medical base station was established at the yellow zone. Two hundred and fourteen passengers were in the trains. Sixty-four of them were injured. They had a median (range) ISS of 2 (1-43), and all were sent to Hospital Kuala Lumpur (HKL). Six (9.4%) patients were clinically triaged as red (critical), 19 (29.7%) as yellow (semi-critical) and 39 (60.9%) as green (non-critical). HKL's disaster plan was activated. All patients underwent temperature and epidemiology link assessment. Seven (10.9%) patients were admitted to the hospital (3 to the ICU, 3 to the ward and 1 to a private hospital as requested by the patient), while the rest 56 (87.5%) were discharged home. Six (9.4%) needed surgery. The COVID-19 tests were conducted on seven patients (10.9%) and were negative. There were no deaths. The mass casualty incident was handled properly because of a clear standard operating procedure, smooth coordination between multi-agencies and the hospitals, presence of a 'binary' system for 'COVID-risk' and 'non-COVID-risk' areas, and the modifications of the existing disaster plan. Preparedness for MCIs is essential during pandemics.


Mitochondrial membrane biogenesis: identification of a precursor to yeast cytochrome c oxidase subunit II, an integral polypeptide.
Many of the polypeptides made on endogenous ribosomes inside of yeast mitochondria are hydrophobic "integral polypeptides" which are subunits of at least three oligomeric enzyme complexes (cytochrome c oxidase, rutamycin-sensitive ATPase, and coenzyme QH2-cytochrome c reductase) of the inner mitochondrial membrane. In order to elucidate the pathway(s) followed by these polypeptides into the inner membrane we have used an in vitro mitochondrial translation system from yeast. By inhibiting this system with aurintricarboxylic acid, we have been able to demonstrate and accumulate a transient precursor to subunit II of cytochrome c oxidase. This precursor, designated II', is approximately 1,500 daltons larger than mature subunit II and most likely is a form of subunit II with an NH2-terminal extension. Although this precursor appears to be processed cotranslationally under normal conditions, it does associate in unprocessed form with mitochondrial membranes when allowed to accumulate in the presence of aurintricarboxylic acid, and it can be processed postranslationally upon removal of the drug. None of the other mitochondrial translation products made in this system exhibits larger precursors. These results indicate that at least one mitochondrial translation product has a transient "leader sequence" a,d is inserted into the inner mitochondrial membrane and processed cotranslationally, but they suggest that other pathways may be followed by the other translation products.


A case of torsion of the wandering spleen presenting as hypersplenism and gastric fundal varices.
Wandering spleen is the displacement of the spleen from its normal location due to the loss or weakening of ligaments that hold the spleen in the left upper quadrant. The possibility of torsion of the spleen is high due to the long and mobile nature of the vascular pedicle. Generally, cases are asymptomatic. Under conditions of delayed diagnosis, symptoms of splenomegaly, left portal hypertension, gastric fundal varices, and hypersplenism may present as a result of development of vascular congestion associated with chronic torsion. There are only a few cases in the literature reporting the association of wandering spleen and fundal varices. We report herein the case of a 55-year-old female who admitted to our clinic with complaints of fatigue and epigastric pain. She was determined to have gastric fundal varices and hypersplenism secondary to the development of left portal hypertension due to chronic splenic torsion.


The use of recombinant activated factor VII in the treatment of massive pulmonary hemorrhage in a preterm infant.
Pulmonary hemorrhage is a rare but well-known complication in preterm infants. We present a case of massive pulmonary hemorrhage in a 9-day-old male infant, successfully treated with intravenous recombinant activated factor VII (rFVIIa) (NovoSeven; Novo Nordisk). The infant was diagnosed with sepsis-related disseminated intravascular coagulation and required ventilator support for respiratory distress syndrome and blood transfusions due to active bleeding from endotracheal tube. After administration of the second dose of rFVIIa (120 microg/kg per dose, every 2 h), the active bleeding subsided dramatically and a significant improvement in the oxygenation index was seen 8 h after the third dose of rFVIIa treatment. There were also significant improvements in the prothrombin time, International Normalized Ratio, activated partial thromboplastin time and plasma fibrinogen levels after the third dose of rFVIIa treatment. The infant was discharged on day 82 of life and there was no finding of thrombosis during the hospitalization period. At month 18 of follow-up, there was no morbidity related to the pulmonary and central nervous systems. This case suggests that rFVIIa is effective as an alternative therapy in controlling massive pulmonary hemorrhage of preterm infants.


HPV-p53-miR-34a axis in HPV-associated cancers.
Human papillomaviruses (HPVs) are known to cause many cancers by altering multiple signalling pathways through their oncogene integration into host genome and expression. Studies have shown that many microRNAs (miRs) may function as oncogenes (called as oncomiRs) to promote an oncogenic effect. MiR-34a among the reported oncomiRs is a key player in the carcinogenesis caused by infection with HPVs. In this mini-review, we summarise the roles of miR-34a in HPV-caused cancers. MiR-34a is transcriptionally regulated by tumour suppressor p53. HPV oncogene E6 inhibits expression of p53 to decrease the levels of miR-34a, leading to the increased expression of multiple genes which are targeted by miR-34a. The upregulation of these genes increases cancer cell proliferation, survival and migration in HPV-associated cancers.


Effects of specific multi-nutrient enriched diets on cerebral metabolism, cognition and neuropathology in AβPPswe-PS1dE9 mice.
Recent studies have focused on the use of multi-nutrient dietary interventions in search of alternatives for the treatment and prevention of Alzheimer's disease (AD). In this study we investigated to which extent long-term consumption of two specific multi-nutrient diets can modulate AD-related etiopathogenic mechanisms and behavior in 11-12-month-old AβPPswe-PS1dE9 mice. Starting from 2 months of age, male AβPP-PS1 mice and wild-type littermates were fed either a control diet, the DHA+EPA+UMP (DEU) diet enriched with uridine monophosphate (UMP) and the omega-3 fatty acids docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), or the Fortasyn® Connect (FC) diet enriched with the DEU diet plus phospholipids, choline, folic acid, vitamins and antioxidants. We performed behavioral testing, proton magnetic resonance spectroscopy, immunohistochemistry, biochemical analyses and quantitative real-time PCR to gain a better understanding of the potential mechanisms by which these multi-nutrient diets exert protective properties against AD. Our results show that both diets were equally effective in changing brain fatty acid and cholesterol profiles. However, the diets differentially affected AD-related pathologies and behavioral measures, suggesting that the effectiveness of specific nutrients may depend on the dietary context in which they are provided. The FC diet was more effective than the DEU diet in counteracting neurodegenerative aspects of AD and enhancing processes involved in neuronal maintenance and repair. Both diets elevated interleukin-1β mRNA levels in AβPP-PS1 and wild-type mice. The FC diet additionally restored neurogenesis in AβPP-PS1 mice, decreased hippocampal levels of unbound choline-containing compounds in wild-type and AβPP-PS1 animals, suggesting diminished membrane turnover, and decreased anxiety-related behavior in the open field behavior. In conclusion, the current data indicate that specific multi-nutrient diets can influence AD-related etiopathogenic processes. Intervention with the FC diet might be of interest for several other neurodegenerative and neurological disorders.


Conformational transition of Escherichia coli RNA polymerase induced by the interaction of sigma subunit with core enzyme.
The isolated sigma subunit of Escherichia coli RNA polymerase has been labeled covalently with a fluorescent probe, N-(1-pyrene)maleimide. The labeled sigma subunit (PM-sigma) still retained its biological activity in stimulating transcription of T7 DNA by core enzyme. When a stoichiometric amount of core enzyme was added to a solution of PM-sigma, there was a decrease in fluorescence intensity without shifts in emission maxima of PM-sigma. The kinetics of the interaction between the sigma subunit and core enzyme was investigated with the stopped-flow technique by monitoring the fluorescence quenching. A biphasic change of fluorescence intensity with respect to time was observed when PM-sigma was rapidly mixed with an excess of core enzyme. The kinetic data can be analyzed in terms of a mechanism in which a fast bimolecular binding of sigma to core enzyme is followed by a relatively slow isomerization of the holoenzyme formed. From the best-fit kinetic parameters, an overall binding constant of less than or equal to 3X10(-10)M was estimated for the PM-sigma core complex, in agreement with that obtained by the fluorimetric titration. In addition, we have studied the effect of temperature on the rate constant associated with the conformational change of the holoenzyme, which shows a temperature transition around 20 degrees C. The nonlinear Arrhenius plot obtained implies that the conformational transition is complex and may be composed of several processes. The activation energy for the "overall" conformational change was estimated to be 6.7 kcal/mol. The kinetic evidence for the conformational transition of holoenzyme induced by the interactions of sigma subunit with core enzyme presented here further supports the proposition that the sigma subunit acts on core enzyme to trap a unique conformation of RNA polymerase which recognizes the proper promoters and initiates the synthesis of specific RNA chains.


Autophagy Enhances Longevity of Induced Pluripotent Stem Cell-Derived Endothelium via mTOR-Independent ULK1 Kinase.
Stem cells are enabling an improved understanding of the peripheral arterial disease, and patient-specific stem cell-derived endothelial cells (ECs) present major advantages as a therapeutic modality. However, applications of patient-specific induced pluripotent stem cell (iPSC)-derived ECs are limited by rapid loss of mature cellular function in culture. We hypothesized that changes in autophagy impact the phenotype and cellular proliferation of iPSC-ECs. Endothelial cells were differentiated from distinct induced pluripotent stem cell lines in 2D culture and purified for CD144 positive cells. Autophagy, mitochondrial morphology, and proliferation were characterized during differentiation and over serial passages in culture. We found that autophagy activity was stimulated during differentiation but stagnated in mature iPSC-ECs. Mitochondria remodeled through mitophagy during differentiation and demonstrated increasing membrane potential and mass through serial passages; however, these plateaued, coinciding with decreased proliferation. To evaluate for oxidative damage, iPSC-ECs were alternatively grown under hypoxic culture conditions; however, hypoxia only transiently improved the proliferation. Stimulating mTOR-independent ULK1-mediated autophagy with a plant derivative AMP kinase activator Rg2 significantly improved proliferative capacity of iPSC-ECs over multiple passages. Therefore, autophagy, a known mediator of longevity, played an active role in remodeling mitochondria during maturation from pluripotency to a terminally differentiated state. Autophagy failed to compensate for increasing mitochondrial mass over serial passages, which correlated with loss of proliferation in iPSC-ECs. Stimulating ULK1-kinase-driven autophagy conferred improved proliferation and longevity over multiple passages in culture. This represents a novel approach to overcoming a major barrier limiting the use of iPSC-ECs for clinical and research applications.


Relationships between cognitive deficits, symptoms and quality of life in schizophrenia.
Schizophrenia is a complex disorder characterized by impairment in a number of domains, all of which contribute to disability. The aim of the present study was to investigate the relationships between cognitive function, symptoms and quality of life (QOL) in schizophrenia. This cross-sectional study measured cognition, positive and negative symptom severity, and quality of life (measured with the Quality of Life Scale) in 57 outpatients with schizophrenia. Correlations between the different measures were sought. Multiple regression analyses were used to develop models of the contributions of cognitive deficits and symptomatology to QOL. More severe positive and negative symptoms and cognitive impairment each correlated with poorer QOL. There was a moderate association between negative symptoms and cognition and a small association between positive symptoms and cognition. Age, gender, and drug and alcohol abuse did not significantly predict QOL. In the multiple regression analysis, entering the total cognition and total symptom scores produced a model that accounted for an additional 57% of the variance in QOL. Improving quality of life for people with schizophrenia requires that positive and negative symptoms and cognition are each addressed as separate domains of impairment. But, given that these account for only 57% of the variance in QOL, other factors such as unemployment, poverty, social isolation and stigma may also be important.


Implant breakage and revision factors for modular fluted tapered stems in revision total hip arthroplasty.
Modular stems in revision total hip arthroplasties allow diaphyseal fixation and optimal restoration of the architecture of the proximal femur. Several studies report metaphyseal implant breakage having a negative impact on survivorship. The aim of the study was to evaluate the outcome of an uncemented modular fluted tapered stem (MFT) in revision surgery. In a retrospective study, 316 patients who had revision surgery using the same design of MFT implant (Modular Revision Stem [MRS], Lima Corporate, Italy) between 2012 and 2017 were identified. Patients were male in 51% of cases and mean age was 74 years. Indications were 110 periprosthetic fractures, 98 periprosthetic joint infection, 97 aseptic loosening, 10 instability and 1 other cause. Survivorship, complications, clinical and radiographic outcomes, were assessed. Mean follow-up was 5 years. There was no implant breakage. At 5-year follow-up, the survivorship, free from revision for aseptic loosening and free of revision for any reason were 96% and 87%, respectively. At last follow-up (8 years), these figures were 92% and 71%, respectively. 31 implants were revised. The risk of revision for any cause was higher for extreme length metaphyseal implants, hazard ratio was 3.7 (95% CI, 1.82-7.52). A mean stem subsidence of 9 mm was noted in 37 cases; 4 were revised for aseptic loosening. The Harris Hip Score at final follow-up was 82. At 5-year follow-up, the MFT implant showed a good survivorship and outcomes with no specific complications. Unlike literature reports, no specific complications occurred with this design. Stem junction positioning and therefore metaphyseal length may be key to optimise long-term survivorship. However, a longer follow-up is needed as implant breakage is more often seen after long implantation times.


Neurotropic and modulatory effects of insulin-like growth factor II in Aplysia.
Insulin-like growth factor II (IGF2) enhances memory in rodents via the mannose-6-phosphate receptor (M6PR), but the underlying mechanisms remain poorly understood. We found that human IGF2 produces an enhancement of both synaptic transmission and neurite outgrowth in the marine mollusk Aplysia californica. These findings were unexpected since Aplysia lack the mammal-specific affinity between insulin-like ligands and M6PR. Surprisingly, this effect was observed in parallel with a suppression of neuronal excitability in a well-understood circuit that supports several temporally and mechanistically distinct forms of memory in the defensive withdrawal reflex, suggesting functional coordination between excitability and memory formation. We hypothesize that these effects represent behavioral adaptations to feeding that are mediated by the endogenous Aplysia insulin-like system. Indeed, the exogenous application of a single recombinant insulin-like peptide cloned from the Aplysia CNS cDNA replicated both the enhancement of synaptic transmission, the reduction of excitability, and promoted clearance of glucose from the hemolymph, a hallmark of bona fide insulin action.


Distribution and content of class 1 integrons in different Vibrio cholerae O-serotype strains isolated in Thailand.
In this study, 176 clinical and environmental Vibrio cholerae strains of different O serotypes isolated in Thailand from 1982 to 1995 were selected and studied for the presence of class 1 integrons, a new group of genetic elements which carry antibiotic resistance genes. Using PCR and DNA sequencing, we found that 44 isolates contained class 1 integrons harboring the aadB, aadA2, blaP1, dfrA1, and dfrA15 gene cassettes, which encode resistance to gentamicin, kanamycin, and tobramycin; streptomycin and spectinomycin; beta-lactams; and trimethoprim, respectively. Each cassette array contained only a single antibiotic resistance gene. Although resistance genes in class 1 integrons were found in strains from the same epidemic, as well as in unrelated non-O1, non-O139 strains isolated from children with diarrhea, they were found to encode only some of the antibiotic resistance expressed by the strains. Serotype O139 strains did not contain class 1 integrons. However, the appearance and disappearance of the O139 serotype in the coastal city Samutsakorn in 1992 and 1993 were associated with the emergence of a distinct V. cholerae O1 strain which contained the aadA2 resistance gene cassette. A 150-kb self-transmissible plasmid found in three O1 strains isolated in 1982 contained the aadB gene cassette. Surprisingly, several strains harbored two integrons containing different cassettes. Thus, class 1 integrons containing various resistance gene cassettes are distributed among different V. cholerae O serotypes of mainly clinical origin in Thailand.


Rare renal pathologic manifestation of antineutrophil cytoplasmic antibodies associated vasculitis as suppurative interstitial nephritis: A case report and review of the literature.
A 69-year old male patient attended our clinic with fatigue, fever, anuria, nephritic syndrome and severe renal failure. Kidney biopsy showed pauci-immune crescentic glomerulonephritis with an unusual association of suppurative interstitial nephritis. Though most patients with renal involvement linked to antineutrophil cytoplasmic antibodies associated vasculitis (AAV) have pauci-immune glomerulonephritis, only a few patients were identified to have atypical renal pathology. In most cases, mononuclear tubulointerstitial infiltrate may be a feature of AAV, suppurative interstitial nephritis is very rare. In the literature, we found only one case reported associated with suppurative interstitial nephritis without glomerulonephritis who later developed classic pauci-immune necrotizing glomerulonephritis. Here, we report a case diagnosed as AAV, presenting with pauci-immune crescentic glomerulonephritis and suppurative interstitial nephritis. It is not clear whether suppurative interstitial nephritis is a severe disease variant in AAV-associated renal disease. As described in the first case the lack of improvement in renal functions in spite of intense immunosuppressive treatment leads to the conclusion that suppurative interstitial nephritis is a marker of poor prognosis.