Acute toxicological study: EZY-1 with potent therapeutic effects of idiopathic pulmonary fibrosis and its mechanisms.
EZY-1 is an antifibrosis peptide purified from Eucheuma. In this study, we explored the acute toxicology of EZY-1 and the signaling pathways involved in its antifibrotic role. The mouse model of pulmonary fibrosis was induced by bleomycin. Pathological changes in lung tissue could be effectively inhibited by EZY-1. Acute toxicity and cell proliferation tests indicated that EZY-1 had no apparent toxicity to mice and cells. We identified proteins that could bind directly to EZY-1 in vitro on the basis of liquid chromatography-tandem mass spectrometry and bioinformatics analysis. EZY-1 inhibited pulmonary fibrosis via Wnt/β-catenin, transforming growth factor (TGF)-β/Smad, phosphoinositide 3-kinase/protein kinase B/ mammalian target of rapamycin, and activator of transcription 3 and Janus kinase 2/signal transducer pathways. A transwell micropore experiment showed that EZY-1 could inhibit cell migration and invasion. Western blotting analysis on transforming growth factor-β1 (TGF-β1)-induced A549 pulmonary fibrosis cell model suggested that EZY-1 could downregulate p-Smad3 (Ser423/Ser425), Smad4, β-catenin, vimentin, and N-cadherin expression. ELISA showed that EZY-1 could inhibit collagen-I secretion. EZY-1 alleviated idiopathic pulmonary fibrosis (IPF) through regulating TGF-β/Smad pathways, epithelial-mesenchymal transition processes, and collagen secretion, which provides a potential foundation for theoretical development of EZY-1 as a potential drug against IPF. PRACTICAL APPLICATIONS: We isolated a new 16-amino-acid peptide derived from the polypeptide extract of Eucheuma, named EZY-1. In vitro and in vivo assays show peptide EZY-1 is safe. The EZY-1 peptide alleviates IPF at lower doses than pirfenidone. EZY-1 alleviated idiopathic pulmonary fibrosis (IPF) through regulating TGF-β/Smad pathways, epithelial-mesenchymal transition (EMT) processes, and collagen secretion, which provides a theoretical basis for the development of EZY-1 as a potential drug against IPF.


Investigation of Methylcyclopentadiene Reactivity: Abstraction Reactions and Methylcyclopentadienyl Radical Unimolecular Decomposition.
Understanding the reactivities of methylcyclopentadiene and the methylcyclopentadienyl radical is important in order to improve our comprehension of the chemical kinetics leading to the formation, decomposition, and growth of the first aromatic ring, as it has been shown that five-membered-ring species are important intermediates in the reaction kinetics of aromatic species. In this work, the rate constants of some key H-abstraction reactions from methylcyclopentadiene to produce the methylcyclopentadienyl radical and the formation of fulvene and benzene from the latter are theoretically determined. Rate constants are evaluated using the ab initio transition state theory-based master equation approach, determining structures and Hessians of all stationary points at the ωB97X-D/aug-cc-pVTZ level, energies at the CCSD(T) level extrapolated to the complete basis set limit, RRKM rate constants using conventional and variational transition state theory, and phenomenological rate constants through the solution of the one-dimensional master equation. Variational corrections are determined in both internal and Cartesian coordinates, and it is found that the choice of the coordinate system can impact the accuracy of the calculated rate constants by up to a factor of 4 for H-abstraction reactions and 2 for the unimolecular decomposition of the methylcyclopentadienyl radical. The calculated rate constants are in good agreement with the available literature data. Prompt dissociation of methylcyclopentadienyl radicals accessed following H-abstraction from methylcyclopentadiene was also investigated, and the corresponding rate constants were determined; the results show that prompt dissociation plays a key role under combustion conditions. Finally, lumping of theoretically derived rate constants to account for methylcyclopentadiene ⇄ methylcyclopentadienyl tautomerism allowed the derivation of a simplified set of rate constants suitable to be inserted into kinetic mechanisms.


Synergistic effect of pasireotide and teriflunomide in carcinoids in vitro.
Somatostatin (SST) analogs are mainstay for controlling tumor proliferation and hormone secretion in carcinoid patients. Recent data suggest that extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation may potentiate the anti-tumor effects of SST analogs in carcinoids. Additionally, ERK1/2 phosphorylating agents have been shown to suppress biomarker expression in carcinoids. Thus, Raf-1/MEK/ERK1/2 pathway activating drugs may be synergistic with SST analogs such as pasireotide (SOM230), which may be more effective than others in its class given its elevated receptor affinity and broader binding spectrum. Here, we investigate the effects of SOM230 in combination with teriflunomide (TFN), a Raf-1 activator, in a human carcinoid cell line. Human pancreatic carcinoid cells (BON) were incubated in TFN, SOM230 or a combination. Cell proliferation was measured using a rapid colorimetric assay. Western analysis was performed to analyze expression levels of achaete-scute complex-like 1 (ASCL1), chromogranin A (CgA), phosphorylated and total ERK1/2, and markers for apoptosis. Combination treatment with SOM230 and TFN reduced cell growth beyond the additive effect of either drug alone. Combination indices (CI) fell below 1, thus quantifiably verifying synergy between both drugs as per the Chou-Talalay CI scale. Combined treatment also reduced ASCL1 and CgA expression beyond the additive effect of either drug alone. Furthermore, it increased levels of phosphorylated ERK1/2, cleaved poly(ADP)-ribose polymerase and caspase-3, and reduced levels of anti-apoptotic biomarkers. Elevated phosphorylated ERK1/2 expression following combination therapy may underlie the synergistic interaction between the two drugs. Since efficacy is achieved at lower doses, combination therapy may palliate symptoms at low toxicity levels. Because each drug has already been evaluated in clinical trials, combinatorial drug trials are warranted.


MRI/PET multimodal imaging of the innate immune response in skeletal muscle and draining lymph node post vaccination in rats.
The goal of this study was to utilize a multimodal magnetic resonance imaging (MRI) and positron emission tomography (PET) imaging approach to assess the local innate immune response in skeletal muscle and draining lymph node following vaccination in rats using two different vaccine platforms (AS01 adjuvanted protein and lipid nanoparticle (LNP) encapsulated Self-Amplifying mRNA (SAM)). MRI and 18FDG PET imaging were performed temporally at baseline, 4, 24, 48, and 72 hr post Prime and Prime-Boost vaccination in hindlimb with Cytomegalovirus (CMV) gB and pentamer proteins formulated with AS01, LNP encapsulated CMV gB protein-encoding SAM (CMV SAM), AS01 or with LNP carrier controls. Both CMV AS01 and CMV SAM resulted in a rapid MRI and PET signal enhancement in hindlimb muscles and draining popliteal lymph node reflecting innate and possibly adaptive immune response. MRI signal enhancement and total 18FDG uptake observed in the hindlimb was greater in the CMV SAM vs CMV AS01 group (↑2.3 - 4.3-fold in AUC) and the MRI signal enhancement peak and duration were temporally shifted right in the CMV SAM group following both Prime and Prime-Boost administration. While cytokine profiles were similar among groups, there was good temporal correlation only between IL-6, IL-13, and MRI/PET endpoints. Imaging mass cytometry was performed on lymph node sections at 72 hr post Prime and Prime-Boost vaccination to characterize the innate and adaptive immune cell signatures. Cell proximity analysis indicated that each follicular dendritic cell interacted with more follicular B cells in the CMV AS01 than in the CMV SAM group, supporting the stronger humoral immune response observed in the CMV AS01 group. A strong correlation between lymph node MRI T2 value and nearest-neighbor analysis of follicular dendritic cell and follicular B cells was observed (r=0.808, P<0.01). These data suggest that spatiotemporal imaging data together with AI/ML approaches may help establish whether in vivo imaging biomarkers can predict local and systemic immune responses following vaccination.


Randomized crossover comparison of right atrial appendage pacing versus interatrial septum pacing for prevention of paroxysmal atrial fibrillation in patients with sinus bradycardia.
New atrial pacing techniques and overdrive pacing algorithms have been introduced to prevent atrial fibrillation. This study was designed to test the hypotheses that (1) interatrial septum pacing (IASP) at the triangle of Koch would be more effective than right atrial appendage pacing (RAAP) in preventing paroxysmal atrial fibrillation (PAF) in patients with sinus bradycardia and (2) an algorithm (CAP) designed to achieve constant atrial capture would increase the efficacy of rate-responsive atrial pacing. We studied 46 patients with PAF and sinus bradycardia implanted with a DDD(R) (Medtronic Thera) pacemaker. Twenty-four patients (6.0 +/- 10.1 PAF episodes/month within 3 months before study) were randomized to RAAP and 22 patients (5.4 +/- 7.1, not significant) to IASP. Within each arm 2 randomized crossover periods of CAP-OFF and CAP-ON function were programed. The PAF episodes per month significantly decreased in the RAAP (CAP-OFF: 2.1 +/- 4.2, P <.05; CAP-ON: 1.9 +/- 3.8, P <.05) and in the IASP group (CAP-OFF: 0.2 +/- 0.5, P <.05; CAP-ON: 0.2 +/- 0.5, P <.05). Values were significantly lower in the IASP group than in the RAAP group in both CAP-OFF (0.2 +/- 0.5 vs 2.1 +/- 4.2, P <.05) and CAP-ON (0.2 +/- 0.5 vs 1.9 +/- 3.8, P <.05) conditions. PAF burden was significantly lower in the IASP than in the RAAP group in CAP-OFF (47 +/- 84 min/d vs 140 +/- 217, P <.05) and in CAP-ON (41 +/- 72 vs 193 +/- 266, P <.05) conditions. No differences were observed within each arm in PAF burden between the 2 crossover CAP programing periods. Rate-adaptive IASP at the triangle of Koch is more effective than RAAP in preventing PAF in patients with sinus bradycardia. In our sample of patients no additional clinical benefit is furnished by the CAP algorithm.


A Sensitive Water-Soluble Reversible Optical Probe for Hg2+ Detection.
We report the serendipitous discovery of an optical mercury sensor while trying to develop a water-soluble manganese probe. The sensor is based on a pentaaza macrocycle conjugated to a hemicyanine dye. The pentaaza macrocycle earlier designed in our group was used to develop photoinduced electron transfer (PET)-based "turn-on" fluorescent sensors for manganese. (1) In an attempt to increase the water-solubility of the manganese sensors we changed the dye from BODIPY to hemicyanine. The resultant molecule qHCM afforded a distinct reversible change in the absorption features and a concomitant visible color change upon binding to Hg2+ ions, leading to a highly water-soluble mercury sensor with a 10 ppb detection limit. The molecule acts as a reversible "ON-OFF" fluorescent sensor for Hg2+ with a 35 times decrease in the emission intensity in the presence of 1 equiv of Hg2+ ions. We have demonstrated the applicability of the probe for detecting Hg2+ ions in living cells and in live zebrafish larvae using confocal fluorescence microscopy with visible excitation. High selectivity and sensitivity toward Hg2+ detection make qHCM an attractive probe for detecting Hg2+ in contaminated water sources, which is a major environmental toxicity concern. We have scrutinized the altered metal-ion selectivity of the probe using density functional theory (DFT) and time-dependent DFT calculations, which show that a PET-based metal-sensing scheme is not operational in qHCM. 1H NMR studies and DFT calculations indicate that Hg2+ ions coordinate to oxygen-donor atoms from both the chromophore and macrocycle, leading to sensitive mercury detection.


Synthesis and characterization of silica gel from Lapindo volcanic mud with ethanol as a cosolvent for desiccant applications.
Lapindo mud (LM) is a volcanic mud from a natural disaster that occurred 16 years ago in Sidoarjo District, East Java, Indonesia. The high amount of silica in the local materials of LM has been extracted for silica gel synthesis via hydrometallurgy methods, followed by sol-gel methods. The presence of ethanol in the synthesis process generated a unique textural property at different ratios between ethanol and sodium silicate (e/ss). Sol-gel mediated silica gel synthesis exhibited mesoporous properties with an amorphous structure, which is a characteristic of the silica gel. The silica gel exhibits silica nanoparticles over the average diameter of 2.08 nm with a spherical morphology and is connected to form an agglomeration structure. Increasing the e/ss ratio enhanced the amount of the hydroxyl group and the specific surface area ranged from 57 to 103 m2 g-1. The moisture adsorption performance of each silica gel showed that the silica gel with an e/ss ratio of 5 : 5 exhibited the highest adsorption capacity measured by conventional gravimetric methods and thermogravimetric analysis of 10.56% and 11.20% gwater gsilica -1, respectively. These results indicated that the silica gel with an e/ss ratio has a high number of hydroxyl groups and more surface-active sites, which is beneficial for the adsorption process. The adsorption capacity of the synthesized silica gel is also higher than that of the commercial silica gel, indicating an excellent performance for desiccant applications.


Identification and characterization of thioredoxin and thioredoxin reductase from Aeropyrum pernix K1.
We have identified and characterized a thermostable thioredoxin system in the aerobic hyperthermophilic archaeon Aeropyrum pernix K1. The gene (Accession no. APE0641) of A. pernix encoding a 37 kDa protein contains a redox active site motif (CPHC) but its N-terminal extension region (about 200 residues) shows no homology within the genome database. A second gene (Accession no. APE1061) has high homology to thioredoxin reductase and encodes a 37 kDa protein with the active site motif (CSVC), and binding sites for FAD and NADPH. We cloned the two genes and expressed both proteins in E. coli. It was observed that the recombinant proteins could act as an NADPH-dependent protein disulfide reductase system in the insulin reduction. In addition, the APE0641 protein and thioredoxin reductase from E. coli could also catalyze the disulfide reduction. These indicated that APE1061 and APE0641 express thioredoxin (ApTrx) and thioredoxin reductase (ApTR) of A. pernix, respectively. ApTR is expressed as an active homodimeric flavoprotein in the E. coli system. The optimum temperature was above 90 degrees C, and the half-life of heat inactivation was about 4 min at 110 degrees C. The heat stability of ApTR was enhanced in the presence of excess FAD. ApTR could reduce both thioredoxins from A. pernix and E. coli and showed a similar molar specific activity for both proteins. The standard state redox potential of ApTrx was about -262 mV, which was slightly higher than that of Trx from E. coli (-270 mV). These results indicate that a lower redox potential of thioredoxin is not necessary for keeping catalytic disulfide bonds reduced and thereby coping with oxidative stress in an aerobic hyperthermophilic archaea. Furthermore, the thioredoxin system of aerobic hyperthermophilic archaea is biochemically close to that of the bacteria.


Constraint and pathology in spontaneously activated systems.
Neural network modeling is approaching a strange attractor. Mathematical demonstrations of the theoretical behavior of non-Lipschitzian dynamical systems claim to show how such self-developing computational networks may be fully deterministic as to their microstructure yet without any prescription of their final state in advance. Justification for these simulations stems from applications which demand more biological realism in the design of neurobiotics. Realization of such modeling in actual hardware does not exist. While the best of these models are surely heuristic for neuroscientists who want to know how brains work, their attempted realization in control systems for planetary high technology seems confounded by several intractable problems, which, a few years ago, would have interested only neurophilosophers in retirement.


Human papilloma virus: a new risk factor in a subset of head and neck cancers.
Head and neck cancer is the sixth most common malignancy worldwide. Tobacco smoking and alcohol consumption are two well known behavioral risk factors associated with head and neck cancer. Recently, evidence is mounting that infection with human papilloma virus, most commonly human papilloma virus-16 is responsible for a subset of head and neck squamous cell carcinoma especially tumors of tonsillar origin. The molecular pathway used by human papilloma virus to trigger malignant transformation of tissue is different from that of other well known risk factors, i.e. smoking and alcohol, associated with squamous cell carcinoma. Apparently, these subsets of patients with human papilloma virus positive tumor are more likely to have a better prognosis than human papilloma virus negative tumor. Considering this fact, the human papilloma virus infection should be determined in all oropharyngeal cancers since it can have a major impact on the decision making process of the treatment.


[Effects of TNF-alpha/NF-kappa B signaling pathway on etanercept alleviating rheumatoid arthritis].
To investigate the effects of etanercept on collagen induced arthritis rats. The rheumatoid arthritis model was established via subcutaneous injection of bovine type Ⅱ collagen, and the effects of etanercept was compared through three groups:①the normal control;②arthritis model treated with saline;③arthritis model treated with etanercept (0.5 mg/kg, intraperitoneal injection) with 10 rats in each group;group②and group③ were selected by arthritis index (≥ 2)and grouped randomly. The body weight and toe volume were measured at just before and during treatment weekly. Rats were sacrificed after 4 weeks of treatment, and the serum levels of tumor necrosis factor α(TNF-α), angiopoietin-1(Ang-1), IL-1β, IL-6 and IL-8 were tested by ELISA. Nuclear factor-kappa B(NF-κB) levels and subcellular locations in toe muscles were test by Western blot. Etanercept significantly relieved inflammation of rheumatoid arthritis, lowered the TNF-α and relative inflammatory factors levels compared with saline treatment (P<0.05). The results of Western blot showed that etanercept significantly suppressed the nuclear location of NF-κB p105/p50 caused by inflammation. The anti-inflammatation effect of etanercept may be related to its activity in inhibiting the activation of TNF-α/NF-κB pathway.


Attributes of clinical role models as described by senior veterinary students in Australia.
Role models incite admiration and provide inspiration, contributing to learning as students aspire to emulate their example. The attributes of physician role models for medical trainees are well documented, but they remain largely unexplored in the context of veterinary medical training. The aim of the current study was to describe the attributes that final-year veterinary students (N=213) at the University of Queensland identified when reflecting on their clinical role models. Clinical role model descriptions provided by students were analyzed using concept-mapping software (Leximancer v. 2.25). The most frequent and highly connected concepts used by students when describing their role model(s) included clients, vet, and animal. Role models were described as good communicators who were skilled at managing relationships with clients, patients, and staff. They had exemplary knowledge, skills, and abilities, and they were methodical and conducted well-structured consultations. They were well respected and, in turn, demonstrated respect for clients, colleagues, staff, and students alike. They were also good teachers and able to tailor explanations to suit both clients and students. Findings from this study may serve to assist with faculty development and as a basis for further research in this area.


Explaining photodermatosis: cyclopentenone vs. alpha-methylene-gamma-lactone natural products.
The possible role of cyclopentenone-containing sesquiterpene lactones in the cause of photochemical chronic actinic dermatitis (CAD) is examined in light of recent reports that the alpha-methylene-gamma-lactone group of these natural products forms 2+2 photoadducts with the DNA base thymine. Neither cyclopentenone nor tenulin (a cyclopentenone-containing sesquiterpene lactone) form such photoadducts with thymine either with sunlight or a UV lamp (300 nm). In contrast, alpha-methylenebutyrolactone readily forms the 2+2 photoadduct with thymine in sunlight. Thus, the photochemical role of the alpha-methylene-gamma-lactone group (rather than cyclopentenone) is strongly implicated in the CAD disease.


Online marketing strategies of plastic surgeons and clinics: a comparative study of the United Kingdom and the United States.
The cosmetic surgery market is a rapidly growing sector of healthcare, and the use of marketing strategies is now an integral part of any cosmetic surgery practice. In this study, the authors review 50 Web sites from practitioners in London and New York to quantify the utilization of online marketing, comparing results between the United Kingdom and the United States.


Cooling evokes redistribution of alpha2C-adrenoceptors from Golgi to plasma membrane in transfected human embryonic kidney 293 cells.
Cold-induced vasoconstriction in cutaneous blood vessels is mediated by increased constrictor activity of vascular alpha2-adrenoceptors (alpha2-ARs). In mouse cutaneous arteries, alpha2-AR constriction at 37 degrees C is mediated by alpha2A-ARs, whereas after cold exposure (28 degrees C), alpha2C-ARs are no longer silent and mediate the remarkable cold-induced augmentation of alpha2-AR responsiveness. The goals of the present study were to develop a cell model of cutaneous thermoregulation and to determine the mechanisms underlying the thermosensitivity of alpha2C-ARs. Human embryonic kidney 293 cells were transiently transfected with the mouse alpha2A- or alpha2C-AR. In cells expressing alpha2A-ARs, UK-14,304 (5-bromo-N-(4,5-dihydro-1H-imidazol-2-yl)-6-quinoxalinamine), an alpha2-AR agonist, inhibited (10 pM) and stimulated (1-10 nM) the accumulation of cAMP evoked by forskolin. Similar responses were obtained at 37 degrees C and 28 degrees C. In contrast, in cells expressing alpha2C-ARs, UK-14,304 did not affect forskolin-stimulated cAMP accumulation at 37 degrees C but did cause a concentration-dependent inhibitory effect at 28 degrees C. Subcellular fractionation revealed that at 37 degrees C alpha2C-ARs were localized predominantly to Golgi compartments, whereas alpha2A-ARs localized predominantly to the plasma membrane. After cooling (28 degrees C), alpha2C-ARs relocated from Golgi compartments to the plasma membrane, whereas the alpha2A-AR remained at the plasma membrane. Immunofluorescence microscopy confirmed that, at 37 degrees C, alpha2A-ARs were localized to the cell surface, whereas alpha2C-ARs colocalized with a trans-Golgi marker. Cooling did not affect localization of alpha2A-ARs, but shifted alpha2C-ARs to the cell surface. Moderate cooling, therefore, caused a selective redistribution of alpha2C-ARs from the Golgi compartments to the cell surface, allowing the rescue of the alpha2C-adrenergic functional response. This mechanism may explain the role of alpha2-ARs in thermoregulation of the cutaneous circulation.


Duck egg-drop syndrome caused by BYD virus, a new Tembusu-related flavivirus.
Since April 2010, a severe outbreak of duck viral infection, with egg drop, feed uptake decline and ovary-oviduct disease, has spread around the major duck-producing regions in China. A new virus, named BYD virus, was isolated in different areas, and a similar disease was reproduced in healthy egg-producing ducks, infecting with the isolated virus. The virus was re-isolated from the affected ducks and replicated well in primary duck embryo fibroblasts and Vero cells, causing the cytopathic effect. The virus was identified as an enveloped positive-stranded RNA virus with a size of approximately 55 nm in diameter. Genomic sequencing of the isolated virus revealed that it is closely related to Tembusu virus (a mosquito-borne Ntaya group flavivirus), with 87-91% nucleotide identity of the partial E (envelope) proteins to that of Tembusu virus and 72% of the entire genome coding sequence with Bagaza virus, the most closely related flavivirus with an entirely sequenced genome. Collectively our systematic studies fulfill Koch's postulates, and therefore, the causative agent of the duck egg drop syndrome occurring in China is a new flavivirus. Flavivirus is an emerging and re-emerging zoonotic pathogen and BYD virus that causes severe egg-drop, could be disastrous for the duck industry. More importantly its public health concerns should also be evaluated, and its epidemiology should be closely watched due to the zoonotic nature of flaviviruses.


Carotid endarterectomy: results and long-term follow-up of a single institution.
To document our experiences, technical procedure and outcomes of carotid endarterectomy (CEA) in patients with symptomatic carotid stenosis. A total of 49 consecutive patients underwent 53 CEAs (three bilateral, one redo) for severe carotid artery stenosis. There were 39 males and 10 females, with mean age of 63 years. All patients underwent neck Doppler, computed tomography/magnetic resonance angiography/digital substraction angiography and a detailed cardiological evaluation. Antiplatelet drugs were continued perioperatively. Surgery was performed under general anesthesia with propofol cerebral protection, mild hypothermia and continuous electroencephalogram monitoring. The procedure was done under microscope and closure was done using 6-0 prolene. Clinical and radiological follow-up was done. Our mean follow-up was 4.69 years. All patients underwent primary suturing except one redo CEA done with venous patch graft. Three patients required intraoperative shunting. One patient died secondary to myocardial infarction peroperatively prior to carotid manipulation. One patient had stroke within 6 h, secondary to operative site intraluminal thrombus and was re-explored. Two patients had transient postoperative hemiparesis and aphasia while two patients had altered sensorium, all self-limiting, with normal imaging. One patient developed temporary twelfth-nerve paresis. One patient had persistent transient ischemic attack on the follow-up. Thus, the perioperative mortality rate was 1.89% and stroke rate was 1.89%. CEA for severe carotid stenosis is a safe procedure with good protection from ischemic events. Detailed preoperative cardiac evaluation and appropriate patient selection is essential.


Stathmin inhibition enhances okadaic acid-induced mitotic arrest: a potential role for stathmin in mitotic exit.
Stathmin is a microtubule-destabilizing phosphoprotein that plays a critical role in the regulation of mitosis. The microtubule-depolymerizing activity of stathmin is lost upon phosphorylation in mitosis. Although the role of phosphorylation of stathmin by p34(cdc2) kinase in the assembly of the mitotic spindle is well established, the role of dephosphorylation of stathmin in mitosis is unknown. In this study, we tested the hypothesis that dephosphorylation of stathmin may be critically important for the depolymerization of the mitotic spindle and the exit from mitosis. We compared the effects of okadaic acid, a specific inhibitor of serine/threonine protein phosphatases, on different parameters of mitotic progression in the presence or absence of stathmin deficiency. Because okadaic acid prevents dephosphorylation of stathmin and results in accumulation of the inactive phosphorylated form, exposure to okadaic acid would be expected to have a more profound effect on mitosis in the presence of relative stathmin deficiency. We found that inhibition of stathmin expression results in increased sensitivity to the antimitotic effects of okadaic acid. This was reflected by increased growth inhibition associated with mitotic arrest. A vast majority of the stathmin-inhibited cells were found to be arrested in late metaphase/anaphase and had severe mitotic spindle abnormalities. Exposure to okadaic acid also resulted in a bigger ratio of polymerized/unpolymerized tubulin in stathmin-inhibited cells relative to control cells. Because the only difference between the control and the stathmin-inhibited cells is the deficiency of stathmin in the latter, the increased susceptibility of the stathmin-inhibited cells to okadaic acid-induced mitotic arrest implies a role for stathmin in the later stages of mitosis.


Caveolin-1 is enriched in the peroxisomal membrane of rat hepatocytes.
Caveolae are a subtype of cholesterol-enriched lipid microdomains/rafts that are routinely detected as vesicles pinching off from the plasma membrane. Caveolin-1 is an essential component of caveolae. Hepatic caveolin-1 plays an important role in liver regeneration and lipid metabolism. Expression of caveolin-1 in hepatocytes is relatively low, and it has been suggested to also reside at other subcellular locations than the plasma membrane. Recently, we found that the peroxisomal membrane contains lipid microdomains. Like caveolin-1, hepatic peroxisomes are involved in lipid metabolism. Here, we analyzed the subcellular location of caveolin-1 in rat hepatocytes. The subcellular location of rat hepatocyte caveolin-1 was analyzed by cell fractionation procedures, immunofluorescence, and immuno-electron microscopy. Green fluorescent protein (GFP)-tagged caveolin-1 was expressed in rat hepatocytes. Lipid rafts were characterized after Triton X-100 or Lubrol WX extraction of purified peroxisomes. Fenofibric acid-dependent regulation of caveolin-1 was analyzed. Peroxisome biogenesis was studied in rat hepatocytes after RNA interference-mediated silencing of caveolin-1 and caveolin-1 knockout mice. Cell fractionation and microscopic analyses reveal that caveolin-1 colocalizes with peroxisomal marker proteins (catalase, the 70 kDa peroxisomal membrane protein PMP70, the adrenoleukodystrophy protein ALDP, Pex14p, and the bile acid-coenzyme A:amino acid N-acyltransferase BAAT) in rat hepatocytes. Artificially expressed GFP-caveolin-1 accumulated in catalase-positive organelles. Peroxisomal caveolin-1 is associated with detergent-resistant microdomains. Caveolin-1 expression is strongly repressed by the peroxisome proliferator-activated receptor-alpha agonist fenofibric acid. Targeting of peroxisomal matrix proteins and peroxisome number and shape were not altered in rat hepatocytes with 70%-80% reduced caveolin-1 levels and in livers of caveolin-1 knockout mice. Caveolin-1 is enriched in peroxisomes of hepatocytes. Caveolin-1 is not required for peroxisome biogenesis, but this unique subcellular location may determine its important role in hepatocyte proliferation and lipid metabolism.


Antileishmanial activity of H1-antihistamine drugs and cellular alterations in Leishmania (L.) infantum.
Leishmaniases are infectious diseases caused by protozoan parasites Leishmania and transmitted by sand flies. Drug repurposing is a therapeutic approach that has shown satisfactory results in their treatment. Analyses of antihistaminic drugs have revealed their in vitro and in vivo activity against trypanosomatids. In this way, this study evaluated the antileishmanial activity of H1-antihistamines and identified the cellular alterations in Leishmania (L.) infantum. Cinnarizine, cyproheptadine, and meclizine showed activity against promastigotes with 50% inhibitory concentration (IC50) values between 10-29 μM. These drugs also demonstrated activity and selectivity against intracellular amastigotes, with IC50 values between 20-35 μM. Fexofenadine and cetirizine lacked antileishmanial activity against both forms. Mammalian cytotoxicity studies revealed 50% cytotoxic concentration values between 52 - >200 μM. These drugs depolarized the mitochondria membrane of parasites and caused morphological alterations, including mitochondrial damage, disorganization of the intracellular content, and nuclear membrane detachment. In conclusion, the L. infantum death may be ascribed by the subcellular alterations followed by a pronounced decrease in the mitochondrial membrane potential, indicating dysfunction in the respiratory chain upon H1-antihistamine treatment. These H1-antihistamines could be used to explore new routes of cellular death in the parasite and the determination of the targets at a molecular level, would contribute to understanding the potential of these drugs as antileishmanial.


[Occurrence and control of psittacosis/ornithosis in West Germany].
Avian chlamydiosis remains to be an important, in birds widely spread zoonosis. In the past 15 years in the Federal Republic of Germany 6133 cases in birds or aviaries officially were recorded, 88% in aviaries consisting of psittacines, 9% in pigeons and 3% in other bird species. Among all bird species psittacines are of greatest importance for human health. Therefore, public health control measures must concentrate on these species. For this purpose the longterm-treatment with tetracyclines in feed or by intramuscular injection has proved effective since chlamydial agents perfectly can be eliminated. But control of psittacosis by medication in the hands of laymen is problematical. Effectivity of medication depends on proper and careful performance. Therefore, if treatment is necessary due to state regulations, strict surveillance, including measuring of antibiotic levels in blood, by experienced veterinary public health personnel is mandatory.


The Molecular Mechanism Underlying Ligand Binding to the Membrane-Embedded Site of a G-Protein-Coupled Receptor.
The crystal structure of P2Y1 receptor (P2Y1R), a class A GPCR, revealed a special extra-helical site for its antagonist, BPTU, which locates in-between the membrane and the protein. However, due to the limitation of crystallization experiments, the membrane was mimicked by use of detergents, and the information related to the binding of BPTU to the receptor in the membrane environment is rather limited. In the present work, we conducted a total of ∼7.5 μs all-atom simulations in explicit solvent using conventional molecular dynamics and multiple enhanced sampling methods, with models of BPTU and a POPC bilayer, both in the absence and presence of P2Y1R. Our simulations revealed that BPTU prefers partitioning into the interface of polar/lipophilic region of the lipid bilayer before associating with the receptor. Then, it interacts with the second extracellular loop of the receptor and reaches the binding site through the lipid-receptor interface. In addition, by use of funnel-metadynamics simulations which efficiently enhance the sampling of bound and unbound states, we provide a statistically accurate description of the underlying binding free energy landscape. The calculated absolute ligand-receptor binding affinity is in excellent agreement with the experimental data (Δ Gb0_theo = -11.5 kcal mol-1, Δ Gb0_exp= -11.7 kcal mol-1). Our study broadens the view of the current experimental/theoretical models and our understanding of the protein-ligand recognition mechanism in the lipid environment. The strategy used in this work is potentially applicable to investigate ligands association/dissociation with other membrane-embedded sites, allowing identification of compounds targeting membrane receptors of pharmacological interest.


Local synchronization in complex networks of coupled oscillators.
We investigate the effects that network topology, natural frequency distribution, and system size have on the path to global synchronization as the overall coupling strength between oscillators is increased in a Kuramoto network. In particular, we study the scenario recently found by Gómez-Gardeñes et al. [Phys. Rev. E 73, 056124 (2006)] in which macroscopic global synchronization emerges through a process whereby many small synchronized clusters form, grow, and merge, eventually leading to a macroscopic giant synchronized component. Our main result is that this scenario is robust to an increase in the number of oscillators or a change in the distribution function of the oscillators' natural frequencies, but becomes less prominent as the number of links per oscillator increases.


Protective effect of thymoquinone, the main component of Nigella Sativa, against diazinon cardio-toxicity in rats.
Several studies have shown that oxidative stress and cell damage can occur at very early stages of diazinon (DZN) exposure. The present study was designed to determine the beneficial effect of thymoquinone (Thy), the main component of Nigella sativa (black seed or black cumin), against DZN cardio-toxicity in rats. In the present experimental study, 48 male Wistar rats were randomly divided into six groups: control (corn oil gavages), DZN gavages (20 mg/kg/day), Thy gavages (10 mg/kg/day) and Thy + DVN gavages (2.5, 5 and 10 mg/kg/day). Treatments were continued for 28 days, then the animals were anesthetized by ether and superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), lactate dehydrogenize (LDH) and glutathione peroxide (GPX) activity was evaluated. In addition, glutathione (GSH) and malondialdehyde (MDA) the heart tissue and creatinephosphokinase-MB (CPK-MB) and troponin (TPI) levels and cholinesterase activity in the blood were evaluated. DZN-induced oxidative damage and elevated the levels of the cardiac markers CK-MB, TPI, MDA and LDH and decreased SOD, CAT and cholinesterase activity and GSH level compared with the control group. Treatment with Thy reduced DZN cardio-toxicity and cholinesterase activity. The success of Thy supplementation against DZN toxicity can be attributed to the antioxidant effects of its constituents. Administration of Thy as a natural antioxidant decreased DZN cardio-toxicity and improved cholinesterase activity in rats through the mechanism of free radical scavenging.


Polypharmacy in Polish Older Adult Population-A Cross-Sectional Study: Results of the PolSenior Project.
Polypharmacy is a challenging issue in geriatrics. The aim of the study was to characterize correlates of polypharmacy in the PolSenior project. The PolSenior project, was a comprehensive survey in a large and longitudinal representative sample of thePolish older population. The project was conducted by the International Institute of Molecular and Cell Biology in Warsaw between 2008 and 2011. All medications consumed during the week preceding the survey were evaluated for each participant (n = 4793, including 2314 females (48.3%)). Thereafter, the percentage of those with polypharmacy (at least 5 medications) and excessive polypharmacy (at least 10 medications) was calculated, and their correlates were determined. The average number of medications used by participants was 5.1 ± 3.6, and was higher in females than in males (5.5 ± 3.5 vs. 4.8 ± 3.5; p < 0.001). Polypharmacy characterized 2650 participants (55.3%) and excessive polypharmacy-532 of them (11.1%). The independent correlates associated withpolypharmacy were: age over 70 years, female sex, higher than primary education, living in an urban area, comorbidities, any hospitalization during past five years, and visiting general practicioners at least yearly. As for correlates with excessive polypharmacy, they were: age 80-84 years, female sex, living in an urban area, diagnosis of at least four chronic diseases, and at least two hospitalizations in the last five years. This study serves as a starting place to understand patient characteristics associated with polypharmacy, excessive polypharmacy, and identify targeted interventions.


A droplet microchip with substance exchange capability for the developmental study of C. elegans.
The nematode Caenorhabditis elegans (C. elegans) has been widely used as a multicellular organism in developmental research due to its simplicity, short lifecycle, and its relevance to human genetics and biology. Droplet microfluidics is an attractive platform for the study of C. elegans in integrated mode with flexibility at the single animal resolution. However, it is still challenging to conduct the developmental study of worms within droplets initiating at the L1 larval stage, due to the small size, active movement, and the difficulty in achieving effective substance exchange within the droplets. Here, we present a multifunctional droplet microchip to address these issues and demonstrate the usefulness of this device for investigating post-embryonic development in individual C. elegans initiating at the larval L1 stage. The key components of this device consist of multiple functional units that enable parallel worm loading, droplet formation/trapping, and worm encapsulation in parallel. In particular, it exhibits superior functions in encapsulating and trapping individual larval L1 worms into droplets in a controlled way. Continuous food addition and expulsion of waste by mixing the static worm-in-droplet with moving medium plugs allows for the long-term culture of worms under a variety of conditions. We used this device to investigate the development processes of C. elegans in transgenic strains with deletion and overexpression of the hypoxia-inducible factor (HIF-1), a highly conserved transcript factor in regulating an organism's response to hypoxia. This microdevice may be a useful tool for the high throughput analysis of individual worms starting at the larval stage, and facilitates the study of developmental worms in response to multiple drugs or environmental toxins.


Coexistence of Autoimmune Disorders and Type 1 Diabetes Mellitus in Children: An Observation from Western Part of India.
Type 1 diabetes mellitus (T1DM) is associated with various autoimmune disorders like celiac disease, thyroid disorder, adrenal failure, etc. However, how common is this association in Indian children is not clearly known. To assess the prevalence of other coexisting autoimmune disorders in children with T1DM. In this cross-sectional study, patients requiring insulin and ketosis-prone diabetic and with history of diabetic ketoacidosis/undetectable fasting C-peptide levels were included. Beside demographic and clinical data, detailed biochemistry evaluations were performed. Celiac disease was diagnosed as per the ESPGHAN diagnostic criteria. ACTH stimulation test was done to confirm the adrenal insufficiency in patients with basal serum cortisol <5 μg/dL. Thyroid function test (TSH) and anti-TPO antibody were assessed in all patients. Screening for other autoimmune disorders was done only when clinically indicated or symptoms or family history was suggestive of presence of such disorder. Among 150 patients enrolled, 64.66% were males and mean age was 13.48 ± 3.29 years (range 3-18 years). Mean age at diagnosis of T1DM was 10.0 ± 3.63 years and duration of diabetes was 3.46 ± 3.18 years. The prevalence of antibodies positive against autoimmune diseases was anti-tTG IgA (20.7%), anti-TPO (33.7%), anti-CCP ab (1.3%), and ANA (0.7%). Significantly higher proportion of females had raised anti-TPO antibodies than males (47.2% vs. 25.8%, P = 0.006). Celiac disease was most common association (24.8%) followed by hypothyroidism (14.1%) and Grave's disease (3.3%). Significantly higher proportion of females had hypothyroidism than males (25.0% vs. 8.2%, respectively, P = 0.005). Prevalence of raised anti-tTG and anti-TPO did not differ significantly by the age (P = 0.841 and P = 0.067) or duration of T1DM (P = 0.493 and P = 0.399). In this part of country, celiac disease, hypothyroidism, and Graves's disease are common associations in children with T1DM.


Cancer services. Left to chance.
Most primary care groups/trusts have cancer lead posts and have some involvement in planning and commissioning cancer services. Cancer is not a high priority in comparison to other national service frameworks and the transition to PCT status. PCG/Ts want help and information about developing cancer services but not all want this now. Most PCG/Ts have some involvement in cancer networks but information needs exist about their role and potential.


Decoronation - a conservative method to treat ankylosed teeth for preservation of alveolar ridge prior to permanent prosthetic reconstruction: literature review and case presentation.
Avulsed teeth that are stored extraorally in a dry environment for >60 min generally develop replacement root resorption or ankylosis following their replantation due to the absence of a vital periodontal ligament on their root surface. One negative sequelae of such ankylosis is tooth infra-positioning and the local arrest of alveolar bone growth. Removal of an ankylosed tooth may be difficult and traumatic leading to esthetic bony ridge deformities and optimal prosthetic treatment interferences. Recently a treatment option for ankylosed teeth named 'de-coronation' gained interest, particularly in pediatric dentistry that concentrated in dental traumatology. This article reviews the up-to-date literature that has been published on decoronation with respect to its importance for future prosthetic rehabilitation followed by a case presentation that demonstrates its clinical benefits.


The associations among sociocultural factors and neuropsychological functioning in older American Indians: The Strong Heart Study.
Valid neuropsychological assessment is critical to the accurate diagnosis and effective treatment of diverse populations. American Indians and Alaska Natives experience substantial health disparities relative to the general U.S. Given the dearth of studies on neuropsychological health in this population, we aimed to characterize neuropsychological performance among older American Indians with respect to age, sex, education, income, and language use. From 2010 to 2014, we recruited 818 American Indians aged 60 and older from the Cerebrovascular Disease and Its Consequences in American Indians Study, who comprised all of the surviving members of a cardiovascular study (Strong Heart Study). This cohort from 11 tribes resided on or near their home reservations in three geographic regions (Northern Plains, Southern Plains, and Southwest). Using a cross-sectional design investigating potential vascular brain injury, we administered a brief, targeted neuropsychological and motor function assessments. Higher scores on neuropsychological tests were associated with younger age, female sex, more education, higher income, and less Native American language use. Similar associations were found for the motor tests, although men had higher scores on both motor function tests. After accounting for other sociocultural and health factors, age, sex, education, income, and Native American language use all had significant associations to the test scores. Our findings may be used to guide research and inform clinical practice. The development of future normative studies for older American Indians will be more culturally appropriate when sociocultural factors are included. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


[Conservative management of adult small bowel intussusception detected at abdominal computed tomography].
The incidence of adult small bowel intussusception detected at CT has increased with advanced imaging techniques and universal utilization of CT scan. We aimed to identify factors that could predict the necessity of surgical intervention in adult patients with small bowel intussusception detected at CT during the past decade. There were 39 cases of adult small-bowel intussusception detected at CT from January 2004 to June 2014. The data on clinical factors, radiological factors and outcomes were collected by retrospectively reviewing all available medical records. Patients were classified as surgical group and conservative group according to the outcome. Association between predictive factors and outcome was assessed by Fisher's exact test and logistic regression models. Among a total of 39 patients, there were 32 patients (82%) in the conservative group and 7 patients (18%) in the surgical group. Spontaneous reduction was confirmed at short-term follow-up studies (abdominal ultrasonography [n=14], single contrast small bowel series [n=14], CT [n=4]) in the conservative group. No recurrence occurred during the median follow-up period of 14.1 months (range, 0-67.5 months). Patients in the surgical group had significantly higher white blood cell (WBC) counts (OR 1.001, p=0.048), more frequent obstruction (n=4 vs. n=4, p=0.022) or leading point (n=5 vs. n=0, p<0.001) and longer intussuception length (OR 1.929, p=0.032). Factors associated with the necessity to resort to surgical intervention in adults with small bowel intussusceptions were higher WBC counts, presence of obstruction or leading point, and longer intussuception length. Conservative management can be considered with short-term follow-up for those without these predictive factors.


Molecular characterization of envelope antigenic variants of hepatitis B virus from Spain.
In a survey of hepatitis B virus (HBV) subtypes using one set of monoclonal typing antibodies, 96.6% of the samples were ad or ay and 3.3% (31) were untypeable. Using two additional antibody panels, 23 samples were ay and ad. Six samples required a third panel to be typed as ay. One sample was not typeable. These last 7 samples were nonreactive or had low reactivity with some antibodies to the common a determinant. Sequencing the HBsAg gene of these 7 samples revealed that amino acid (aa) 122 was arginine, as expected for the y determinant. However, mutations giving rise to serine at aa 120, leucine at aa 143, and glutamic acid at aa 144 were found singly or in combination and correlated with the monoclonal antibody binding patterns. These results have implications for further subtyping surveys, vaccines, immunotherapy, and the design of diagnostic assays, and they give clues to the structure of the major neutralizing epitope of HBV.


Role of p38 mitogen-activated protein kinase in chemokine-induced emigration and chemotaxis in vivo.
It has been proposed that L-selectin engagement with ligand activates p38 mitogen-activated protein kinase (MAPK) and can impact on downstream events of leukocyte rolling, including adhesion, and emigration. Using a novel chemotactic assay in vivo, we visualized slow release of chemokine from an agarose gel positioned 350 microm from a postcapillary venule, which induced directed migration (chemotaxis) of neutrophils. In this system, keratinocyte-derived cytokine induced phosphorylation of p38 MAPK, which phosphorylated a downstream protein (ATF-2). This latter event was blocked by the concentration of p38 inhibitors used in this study. Mice were treated with two different p38 inhibitors: SKF86002 and SB203580. Neither inhibitor affected rolling or adhesion in microvessels. Intravenous treatment with SFK86002 (5, 10, and 20 mg/kg) 30 min before the inflammatory stimulus inhibited the total number of emigrated cells at a dose of 20 mg/kg (62%, p < 0.05), despite the presence of many adherent cells within the vessels. A similar inhibition was observed with 20 mg/kg of a second p38 inhibitor SB203580 (67%, p < 0.05). In addition to emigration, both p38 inhibitors impaired the ability of emigrated cells to migrate through the tissue toward the chemotactic stimulus. In fact, the majority of emigrated leukocytes in p38 inhibitor-treated animals remained within 50 microm of the venule. Superfusion of the tissue with SKF86002 (0.7 mM) to impact only on emigrated and not vascular leukocytes resulted in no impairment in emigration, but in a significant reduction in chemotaxis away from the vessel wall. Again, the majority of emigrated leukocytes remained within 50 microm of the blood vessel. Our results suggest that p38 does not affect rolling or adhesion, but that it is involved in leukocyte emigration and chemotaxis through interstitium in response to keratinocyte-derived cytokine in vivo.


Evaluation of Mechanical and Electrical Performance of Aging Resistance ZTA Composites Reinforced with Graphene Oxide Consolidated by SPS.
This paper presents a study of Al2O3-ZrO2 (ZTA) nanocomposites with different contents of reduced graphene oxide (rGO). The influence of the rGO content on the physico-mechanical properties of the oxide composite was revealed. Graphene oxide was obtained using a modified Hummers method. Well-dispersed ZTA-GO nanopowders were produced using the colloidal processing method. Using spark plasma sintering technology (SPS), theoretically dense composites were obtained, which also reduced GO during SPS. The microstructure, phase composition, and physico-mechanical properties of the sintered composites were studied. The sintered ZTA composite with an in situ reduced graphene content of 0.28 wt.% after the characterization showed improved mechanical properties: bending strength was 876 ± 43 MPa, fracture toughness-6.8 ± 0.3 MPa·m1/2 and hardness-17.6 ± 0.3 GPa. Microstructure studies showed a uniform zirconia distribution in the ZTA ceramics. The study of the electrical conductivity of reduced graphene oxide-containing composites showed electrical conductivity above the percolation threshold with a small content of graphene oxide (0.28 wt.%). This electrical conductivity makes it possible to produce sintered ceramics by electrical discharge machining (EDM), which significantly reduces the cost of manufacturing complex-shaped products. Besides improved mechanical properties and EDM machinability, 0.28 wt.% rGO composites demonstrated high resistance to hydrothermal degradation.


Thymoquinone attenuates phosphorylation of AKT to inhibit kidney cancer cell proliferation.
Thymoquinone (Tq) is an active compound from Nigella sativa which is used in traditional medicine. The effect of Tq on kidney cancer and the pathway of action remain unproven. Herein, we report the anticancer properties of Tq on kidney cancer cells. Tq demonstrated anti-proliferative effects in A498 cells with a GI50 value of 40.07 µM and Caki-1 cells with a GI50 of 51.04 µM by the MTT assay. Tq exhibited nuclear fragmentation and inhibited trans-endothelial migration of A498 and Caki-1 cells in a dose-responsive manner. Time-dependent increase in Annexin V-positive cells and sub-G0 /G1 cell population was observed post-Tq treatment. The compound increased Bax protein levels and reduced Bcl-2 protein levels dose dependently in cells, thereby favoring apoptosis. Tq decreased the phosphorylation of Akt in both kidney cell types. The results suggest effective anticancer activity of Tq on kidney cancer cells which may be mediated by the Akt pathway. PRACTICAL APPLICATIONS: Results from the current investigation will through more light on the mechanistic pathway of Tq activity on the inhibition of kidney cancer cell proliferation. The output of this preclinical in vitro study may be translated into better chemotherapeutics of Tq and its analogs to treat kidney cancer. However, a detailed investigation on in vivo models is recommended.


[Usefulness of M-mode echocardiography, two-dimensional and Doppler, in the study of pericardial diseases].
The objective of this communication is to determine utility of echocardiography M mode, two-dimensional and Doppler, in the diagnosis and follow-up of patients with pericardial disease. This procedure was used initially to the diagnosis of pericardial effusion, actually it has a lot of new applications. It's noninvasive and low cost, and would give us important information about our patients.


Mining for Oxysterols in Cyp7b1-/- Mouse Brain and Plasma: Relevance to Spastic Paraplegia Type 5.
Deficiency in cytochrome P450 (CYP) 7B1, also known as oxysterol 7α-hydroxylase, in humans leads to hereditary spastic paraplegia type 5 (SPG5) and in some cases in infants to liver disease. SPG5 is medically characterized by loss of motor neurons in the corticospinal tract. In an effort to gain a better understanding of the fundamental biochemistry of this disorder, we have extended our previous profiling of the oxysterol content of brain and plasma of Cyp7b1 knockout (-/-) mice to include, amongst other sterols, 25-hydroxylated cholesterol metabolites. Although brain cholesterol levels do not differ between wild-type (wt) and knockout mice, we find, using a charge-tagging methodology in combination with liquid chromatography-mass spectrometry (LC-MS) and multistage fragmentation (MSn), that there is a build-up of the CYP7B1 substrate 25-hydroxycholesterol (25-HC) in Cyp7b1-/- mouse brain and plasma. As reported earlier, levels of (25R)26-hydroxycholesterol (26-HC), 3β-hydroxycholest-5-en-(25R)26-oic acid and 24S,25-epoxycholesterol (24S,25-EC) are similarly elevated in brain and plasma. Side-chain oxysterols including 25-HC, 26-HC and 24S,25-EC are known to bind to INSIG (insulin-induced gene) and inhibit the processing of SREBP-2 (sterol regulatory element-binding protein-2) to its active form as a master regulator of cholesterol biosynthesis. We suggest the concentration of cholesterol in brain of the Cyp7b1-/- mouse is maintained by balancing reduced metabolism, as a consequence of a loss in CYP7B1, with reduced biosynthesis. The Cyp7b1-/- mouse does not show a motor defect; whether the defect in humans is a consequence of less efficient homeostasis of cholesterol in brain has yet to be uncovered.


Effect of chronic morphine administration on glucose tolerance and insulin binding to isolated rat adipocytes.
Morphine addiction was induced in six male Wistar rats. Improved glucose tolerance (peak value less by 28%, p<0.01) was observed in chronically morphinized rats as compared to the control rats, injected with saline. An increase in the maximal specific binding of 125I-labeled insulin to unit membrane area of adipocytes was observed in the experimental group (p < 0.01). The changes in insulin receptor number could be responsible for the improved glucose tolerance observed during morphine addiction.


Temporal expression of transcription and relative copy number of plasmid pSFB-1 in Scytalidium flavo-brunneum.
Wild-type cultures of Scytalidium flavo-brunneum produce a 15-azasterol antifungal agent and a reddish brown pigment as secondary metabolites. Spontaneous mutants of S. flavo-brunneum that had simultaneously lost the ability to produce the 15-azasterol and the pigment were transformed with plasmid pSFB-1, which was obtained from wild-type S. flavo-brunneum. Each transformant possessed the plasmid and coincidentally reacquired azasterol and pigment production. Regulation of transcription and relative plasmid copy number was determined as a function of the culture cycle of the organism. Twenty-five-fold amplification of the plasmid was observed in the fungus during the stationary phase. RNA transcripts of 0.9, 1.0, 1.5, 2.0, and 2.5 kilobases were expressed from the plasmid by the organism. While differences in the temporal regulation of the transcripts were seen, all except the 1.0-kilobase transcript increased in abundance on entry of the culture into the stationary phase of growth.


Subject-specific finite element models can accurately predict strain levels in long bones.
The prediction of the stress-state and fracture risk induced in bones by various loading conditions in individual patients using subject-specific finite element models still represents a challenge in orthopaedic biomechanics. The accuracy of the strain predictions reported in the literature is variable and generally not satisfactory. The aim of the present study was to evaluate if a proper choice of the density-elasticity relationship can lead to accurate strain predictions in the frame of an automatic subject-specific model generation strategy. To this aim, a combined numerical-experimental study was performed comparing finite element predicted strains with strain-gauges measurements obtained on eight cadaver proximal femurs, each instrumented with 15 rosettes mostly concentrated in the bone metaphyses, tested non-destructively in vitro under six different loading scenarios. Three different density-elasticity power relationships were selected from the literature and implemented in the finite element models derived from computed tomography data. The results of the present study confirm the great influence of the density-elasticity relationship used on the accuracy of numerical predictions. One of the tested constitutive laws provided a very good agreement (R(2)=0.91, RMSE lower than 10% of the maximum measured value) between numerical calculations and experimental measurements. The presented results show, in addition, that the adoption of a single density-elasticity relationship over the whole bone density range is adequate to obtain an accuracy that is already suitable for many applications.


Interaction of L-lysine and soluble elastin with the semicarbazide-sensitive amine oxidase in the context of its vascular-adhesion and tissue maturation functions.
The copper-containing quinoenzyme semicarbazide-sensitive amine oxidase (EC 184.108.40.206; SSAO) is a multifunctional protein. In some tissues, such as the endothelium, it also acts as vascular-adhesion protein 1 (VAP-1), which is involved in inflammatory responses and in the chemotaxis of leukocytes. Earlier work had suggested that lysine might function as a recognition molecule for SSAO/VAP-1. The present work reports the kinetics of the interaction of L-lysine and some of its derivatives with SSAO. Binding was shown to be saturable, time-dependent but reversible and to cause uncompetitive inhibition with respect to the amine substrate. It was also specific, since D-lysine, L-lysine ethyl ester and epsilon-acetyl-L-lysine, for example, did not bind to the enzyme. The lysine-rich protein soluble elastin bound to the enzyme relatively tightly, which may have relevance to the reported roles of SSAO in maintaining the extracellular matrix (ECM) and in the maturation of elastin. Our data show that lysyl residues are not oxidized by SSAO, but they bind tightly to the enzyme in the presence of hydrogen peroxide. This suggests that binding in vivo of SSAO to lysyl residues in physiological targets might be regulated in the presence of H(2)O(2), formed during the oxidation of a physiological SSAO substrate, yet to be identified.


A portable total reflection X-ray fluorescence spectrometer with a diamond-like carbon coated X-ray reflector.
A diamond-like carbon (DLC) coated quartz glass sample holder is used in a portable total reflection X-ray fluorescence (TXRF) spectrometer. A spot area of a water sample on the DLC sample holder becomes smaller than that on a quartz glass sample holder usually used in TXRF analysis because DLC is more hydrophobic. Therefore, the use of the DLC sample holder enhances fluorescent X-rays reaching a detector compared with the use of a quartz glass sample holder, leading to improvement in detection sensitivity. A detection limit of 28 pg is achieved for Cr in a river water sample when using the DLC sample holder.


Gastroprotective effect of apricot kernel oil in ethanol-induced gastric mucosal injury in rats.
We investigated the gastroprotective effect of apricot kernel oil on ethanol induced gastric ulcer in rats. Male Wistar albino rats were divided into control, ethanol and apricot kernel oil + ethanol groups. The fatty acid composition of apricot kernel oil was determined using GC-MS. A gastric ulcer index was defined as the area percentage of the gastric mucosa consisting of ulcerated tissue. Gastric tissue was investigated by TUNEL staining for apoptosis, immunohistochemical iNOS staining, measurement of gastric IL-10 and IL-6 expression by ELISA and assays of catalase, malondialdehyde and superoxide dismutase. The ethanol group exhibited a higher gastric ulcer score, increased IL-6 level, increased number of inducible nitric oxide synthase-positive and TUNEL positive cells, and a higher MDA level compared to the control group. The apricot kernel oil + ethanol group exhibited significantly fewer gastric lesions compared to the ethanol group. Apricot kernel oil protects rat gastric mucosa against ethanol induced injury by its anti-inflammatory, anti-oxidative and anti-apoptotic effects, and might be useful for reducing the severity of gastric ulcers.


Maximum inspiratory pressure, a surrogate parameter for the assessment of ICU-acquired weakness.
Physical examination has been advocated as a primary determinant of ICU-acquired weakness (ICU-AW). The purpose of the study is to investigate ICU-AW development by using Maximum Inspiratory Pressure (MIP) as a surrogate parameter of the standardized method to evaluate patients' peripheral muscle strength. Seventy-four patients were recruited in the study and prospectively evaluated in a multidisciplinary university ICU towards the appearance of ICU-AW. APACHE II admission score was 16 ± 6 and ICU stay 26 ± 18 days. ICU-AW was diagnosed with the Medical Research Council (MRC) scale for the clinical evaluation of muscle strength. MIP was measured using the unidirectional valve method, independently of the patients' ability to cooperate. A significant correlation was found between MIP and MRC (r = 0.68, p < 0.001). Patients that developed ICU-AW (MRC<48) had a longer weaning period compared to non ICU-AW patients (12 ± 14 versus 2 ± 3 days, p < 0.01). A cut-off point of 36 cmH2O for MIP was defined by ROC curve analysis for ICU-AW diagnosis (88% sensitivity,76% specificity). Patients with MIP below the cut-off point of 36 cmH2O had a significant greater weaning period (10 ± 14 versus 3 ± 3 days, p = 0.004) also shown by Kaplan-Meier analysis (log-rank:8.2;p = 0.004). MIP estimated using the unidirectional valve method may be a potential surrogate parameter for the assessment of muscle strength compromise, useful for the early detection of ICU-AW.


Synthesis of an Undecasaccharide Featuring an Oligomannosidic Heptasaccharide and a Bacterial Kdo-lipid A Backbone for Eliciting Neutralizing Antibodies to Mammalian Oligomannose on the HIV-1 Envelope Spike.
Lipooligosaccharides (LOS) from the bacterium Rhizobium radiobacter Rv3 are structurally related to antigenic mammalian oligomannoses on the HIV-1 envelope glycoprotein spike that are targets for broadly neutralizing antibodies. Here, we prepared a hybrid structure of viral and bacterial epitopes as part of a vaccine design strategy to elicit oligomannose-specific HIV-neutralizing antibodies using glycoconjugates based on the Rv3 LOS structure. Starting from a Kdo2GlcNAc2 tetrasaccharide precursor, a central orthogonally protected mannose trichloroacetimidate donor was coupled to OH-5 of the innermost Kdo residue. To assemble larger glycans, the N-acetylamino groups of the glucosamine units were converted to imides to prevent formation of unwanted imidate byproducts. Blockwise coupling of the pentasaccharide acceptor with an α-(1→2)-linked mannotriosyl trichloroacetimidate donor introduced the D1-arm fragment. Glycosylation of O-6 of the central branching mannose with an α-(1→2)-α-(1→6)-linked mannotriosyl trichloroacetimidate donor unit then furnished the undecasaccharide harboring a D3-arm extension. Global deprotection yielded the 3-aminopropyl ligand, which was activated as an isothiocyanate or adipic acid succinimidoyl ester and conjugated to CRM197. However, representative oligomannose-specific HIV-neutralizing antibodies bound the undecasaccharide conjugates poorly. Possible reasons for this outcome are discussed herein along with paths for improvement.


Enlarged accessory spleens: CT appearance in postsplenectomy patients.
An unexplained abdominal mass discovered on CT examinations of a postsplenectomy patient may represent a hypertrophied residual accessory spleen. Six such patients were found to have left upper quadrant masses on CT examinations that were subsequently proved to be enlarged accessory spleens. The CT appearance is that of discrete, solitary, rounded masses of uniform soft-tissue density in the left upper quadrant. Diameters of the masses were 3.5-5.0 cm. The splenic nature of such a mass can be confirmed by appropriate radionuclide imaging.


Progesterone for preventing premature birth: practice patterns of board-certified maternal-fetal medicine specialists in the United States.
To determine the current prescription of progesterone to prevent preterm birth (PTB) among board-certified maternal-fetal medicine (MFM) specialists in the United States. A survey of the board-certified MFM specialists in the United States examining their prescription of and attitudes regarding progesterone to prevent PTB 6 months following publication of a National Institute for Child Health and Human Development trial. Of 1,264 questionnaires sent, 526 were returned (response rate, 42%). After exclusions, 522 surveys remained. One hundred ninety-eight (38%) respondents prescribed progesterone, and 324 (62%) did not. Most nonprescribers were awaiting more data and were more concerned than prescribers about long-term effects (p < 0.0001). Twenty percent of prescribers prescribed progesterone for women with current signs or symptoms of preterm labor. As a result of recent evidence, over one third of MFM specialists surveyed have begun prescribing progesterone to prevent PTB. Of these specialists, 20% are using it for indications other than a prior PTB.


[Protein post-translational modification and age-related cataract].
Cataract is the main cause of vision impairment, and age-related cataract is the most common type of cataract. Protein post-translational modification is the enzymatic processing of a polypeptide chain after translation from messenger RNA and after peptide bond formation has occurred. Deamidation, glycation and oxidation are age-related processes. They can change the structure or solubility of crystallin and result in the opacity of lens while acetylation and methylation may be related to the protection of lens protein. Although much effort has been directed towards slowing progression or preventing the occurrence of cataract, the management of cataract remains surgical. This review deals with the relations between different kinds of protein post-translational modification and age-related cataract as well as some protective methods aiming at the process of post-translational modification.


Mass Spectrometric DNA Adduct Quantification by Multiple Reaction Monitoring and Its Future Use for the Molecular Epidemiology of Cancer.
The formation of DNA adducts is considered essential for tumor initiation. Quantification of DNA adducts may be achieved by various techniques of which LC-MS/MS-based multiple reaction monitoring has become the most prominent in the past decade. Adducts of single nucleosides are analyzed following enzymatic break-down of a DNA sample following adduct enrichment usually by solid-phase extraction. LC-MS/MS quantification is carried out using stable isotope-labeled internal reference substances. An upcoming challenge is the use of DNA adducts as biomarkers either for internal exposure to electrophilic genotoxins or for the approximation of cancer risk. Here we review recent studies in which DNA adducts were quantified by LC-MS/MS in DNA samples from human matrices. We outline a possible way for future research to aim at the development of an 'adductome' approach for the characterization of DNA adduct spectra in human tissues. The DNA adduct spectrum reflects the inner exposure of an individual's tissue to electrophilic metabolites and, therefore, should replace the conventional and inaccurate external exposure in epidemiological studies in the future.


Human papillomavirus infection in malignant and benign gynaecological conditions: a study in Greek women.
HPV infection is by far the most frequent sexually transmitted disease. Our aim in this prospective nonrandomized study was to investigate the frequency with which different subtypes of the human papillomavirus (HPV) are found in gynaecological malignant and benign conditions and to compare the rate of infection between them. Detailed data of 195 women were selected and divided into three groups. The first group consisted of specimens from 68 women with cervical cancer. The second group consisted of specimens from 43 women with endometrial and ovarian cancer. The third group consisted of 84 specimens from women who were operated on for benign gynaecological diseases. Seven oncogenic types of HPV (6, 11 [low-risk subtypes] 16, 18, 31, 33, and 51 [high-risk subtypes]) were investigated by using the in situ hybridization technique. The HPV detection rate was higher in the cervical cancer group (74.8%), compared with the second (27.9%) and third (45.2%) groups. The most common HPV subtypes in the first group were 16 and 18. The most common subtypes in the second group were the 31, 33, and 51, and in the third group the 6 and 11 subtypes. Women developed cervical cancer at a younger age than endometrial or ovarian cancer. Smoking and exposure to multiple sexual partners appeared related to the development of cervical cancer. The use of contraceptive pill, dietary habits, and diabetes did not seem to increase the risk of HPV infection in this population. As it is common in other populations, in this selected population of Greek women we were able to confirm that in women with cervical cancer the most common HPV subtypes are 16 and 18. Education of adolescents on the epidemiological association of smoking and unprotected intercourse with cervical cancer will certainly help to reduce the rates of development of preinvasive and invasive cancer of the cervix in young women.


Endoscopic management of duodenal adenomas in familial adenomatous polyposis--a single-center experience.
Duodenal lesions (DLS) are common in patients with familial adenomatosis polyposis (FAP), and screening for duodenal adenocarcinoma (DA) is currently recommended. Endoscopic treatment of DLS is controversial. To report management and outcomes of endoscopic therapy for DLS in patients with FAP. The records of patients with FAP who underwent endoscopic surveillance or therapy for DLS over a 15-year period were reviewed. Endoscopic intervention included endoscopic surveillance with biopsies, argon plasma coagulation (APC), endoscopic mucosal resection (EMR), EMR with APC, and ampullectomy. Main outcome measurements were recurrence and histology of DLS after endoscopic therapy, complications of endoscopic therapy, and need for duodenectomy. Seventy-one patients with FAP and DLS were identified from our endoscopy database as undergoing upper endoscopy for screening and/or surveillance (1995-2009). Mean follow up was 4.5 years (1-15 years). Seventy of the seventy-one (98.5%) patients had multiple flat DLS. Most of the patients were followed with yearly biopsies. APC was performed in 17 patients and EMR was performed in eight patients; in five of the eight EMR patients, APC was also performed to treat the edges of EMR site. During the follow up, 17/55 (31%) patients had histological progression (HP). HP was seen in 5/16 (31%) patients who underwent APC (one was lost to follow-up) and 12/40 (30%) patients followed with biopsies alone. Recurrence of lesions was noted in all patients. Two patients underwent duodenectomy. None of the patients developed DA during follow up. Endoscopic surveillance with directed endotherapy for DLS in FAP is feasible and safe when diligently performed.


The clinical characteristics, therapeutic outcome, and prognostic factors of non-tuberculous bacterial spinal epidural abscess in adults: a hospital-based study.
Nontuberculous bacterial spinal epidural abscess (SEA) is a serious infectious disease of spines. Despite proper management, SEA remains a disease of high morbidity. The epidemiological data in southern Taiwan is still lacking. We analyzed the clinical features, laboratory data, magnetic resonance imaging findings, and therapeutic results of 45 bacterial SEA patients, collected during a study period of six years (2003- 2008). The 45 adult SEA patients, 32 were men and 13 women, aged 17-81 years (mean = 62 years). Thoracic and lumbosacral spines were the most commonly affected segments. Staphylococcus aureus was the most commonly implicated pathogen, accounting for infections in 67% (29/43) of the culture-proven cases, and 41% (12/29) of the S. aureus strains were oxacillin resistant. Gram-negative pathogens were implicated in 30% (13/43) of the culture-proven cases. Of the 45 patients, 19 underwent antibiotic treatment alone and 26 underwent medical and surgical treatments. The therapeutic results showed that 28 patients had a good prognosis and 17 had a poor prognosis. The significant prognostic factors included higher age, presence of diabetes mellitus and chronic heart disease, and initial presentation of altered consciousness. S. aureus was the most commonly implicated pathogen of adult SEA, and most S. aureus strains were oxacillin resistant. A relatively high incidence of Gram-negative infection was observed in this study. The epidemiological characteristics of the implicated pathogens must receive special consideration when selecting the empirical antibiotics. Despite adequate management, many adults with bacterial SEA had a high incidence of poor prognosis.


Is allogeneic blood transfusion a risk factor for sternal dehiscence following cardiac surgery? A prospective observational study.
Sternal dehiscence following cardiac surgery has a multifactorial etiology. Significant risk factors contributing to sternal dehiscence include chronic obstructive pulmonary disease (COPD), obesity, or re-exploration due to bleeding or pericardial tamponade. We have focused on the role of allogeneic blood transfusion as a factor leading to the poor healing of surgical wounds. A prospective observational study of 1553 elective and emergency cardiac surgery patients was performed between January 2003 and June 2007. All of the patients enrolled in this study underwent median sternotomy. We studied the relationship between sternal dehiscence following cardiac surgery and the total number of packed red blood cells transfused. The incidence of sternal dehiscence in the study group was 3.4%. Diabetic patients did not have a higher incidence of wound dehiscence. Although COPD, obesity, and re-exploration contributed to sternal dehiscence, the number of allogeneic blood transfusions during the perioperative period was an important independent risk factor for sternal dehiscence. Patients with sternal dehiscence received an average of 7.6 transfusion units (TU) of allogeneic blood versus 1.6 TU of allogeneic blood in the group without sternal dehiscence (P < 0.00005). The dehiscence affected patients without any other significant risk factor who received 6 or more TU, or patients with at least one significant risk factor who received 4 or more TU of allogeneic blood. According to our results, the total amount of allogeneic blood transfused is an important risk factor contributing to sternal dehiscence. Regardless of other risk preconditions, the transfusion of 6 or more TU could result in sternal dehiscence following cardiac surgery.


[Synthetic suburethral sling in the treatment of stress urinary incontinence in women].
Since the marketing and success of the TVT technique, other products have been released onto the suburethral sling market. These products can be classified according to four main criteria: TAPE: The material used in every case is polypropylene, in the form of woven monofilaments (TVT, Sparc, Monarc, Uretex Sup), multi-filaments (IVS, Safyre comprising two silicone tension bands), extruded crosslinked polypropylene (Obtape) or silicone-coated polypropylene in contact with the urethra (Uratape). A larger space between the fibres is associated with a lower theoretical risk of infection and better tissue ingrowth. The weaving process of Uretex Sap tape decreases the warping and release effect. Extruded polypropylene forms a very solid, but poorly elastic tape. It is not yet clear whether silicone-coating is an advantage or a disadvantage. The ascending retropubic approach (TVT, Uretex Sup, IVS) is now well known. It carries a risk of bladder perforation and, more exceptionally, intestinal or vascular injuries. The descending retropubic approach (Uretex Sup, Sparc) requires more extensive vaginal dissection and carries a risk of bladder perforation. The mixed retropubic approach (Uretex Sup) combines the potential disadvantages of the ascending and descending approaches. The transobturator approach (Uratape, Obtape, Monarc) was described more recently and is easy to learn. It does not carry any risk of intestinal or vascular injury, but the urethra, vagina or even the bladder (in the case of lateral cystocele) can be damaged by this incision. Ease of use is an essential feature. Uretex Sup comprises ancillary instruments and a technique requiring multiple manipulations. Sparc tape cannot be easily dissociated from the ancillary instruments when a second passage is necessary. TVT and Uratape use resterilizable instruments, while the other products are totally disposable. FOLLOW-UP AND PUBLICATIONS: Only TVT has a sufficient follow-up and several hundred specific papers or publications. Only a few publications by a few different teams are currently available for the other products. In conclusion, the various published studies confirm the qualities and good tolerance of woven monofilament polypropylene. The other materials require a longer follow-up and more studies before they can be completely validated in terms of results and morbidity. The multiple surgical approaches and the various ancillary instruments available allow the technique to be adapted to each patient and each surgeon, regardless of his or her initial training and habits.


The Rorschach and MMPI: toward a more scientifically differentiated understanding of cross-method assessment.
Reasons for Minnesota Multiphasic Personality Inventory (MMPI) and Rorschach disagreement at the nomothetic level are explored. Building on an understanding of measurement distinctions from other sciences, it is proposed that the Rorschach and MMPI procedures are differentially sensitive to unique manifestations of personality. By necessity, each method is then also recognized as having a limited scope of effectiveness, such that neither can provide a complete picture of personality in its full complexity. Drawing on the more extensive self-report literature, the idiosyncracies and limited scope of the self-report method are documented. Finally, an ideographically rooted, cross-method, configural model is proposed for validation research in personality assessment. Several examples consistent with this approach are drawn from the assessment literature and discussed.


Transesophageal echocardiography impacts management and evaluation of patients with stroke, transient ischemic attack, or peripheral embolism.
The relative impact of transesophageal echocardiography (TEE) on the management of patients with specific embolic events, namely nonhemorrhagic cerebrovascular accident (CVA), transient ischemic attack (TIA), or peripheral embolism is controversial. The impact of TEE in 234 adult subjects with CVA (n = 141), TIA (n = 59), or peripheral embolism (n = 34) was determined. TEE was diagnostic of a potential embolic source in 61%, 51%, and 62% of patients with CVA, TIA, and peripheral embolism, respectively (P = NS). TEE results changed medication or surgical treatment in 32%, 22%, and 32% of patients with CVA, TIA, and peripheral embolism, respectively (P = NS). Anticoagulation was started on the basis of TEE findings in 11%, 12%, and 18% of patients with CVA, TIA, and peripheral embolism, respectively (P = NS). In 77% of all patients, TEE findings confirmed as appropriate the empiric decision made prior to TEE, to anticoagulate (60%; 12/20) or not to anticoagulate (79%; 168/214). These data demonstrate that TEE findings have a significant and similar impact on the clinical management of patients with various types of potential embolism. Future studies addressing the effectiveness of treatment, guided by TEE findings, in the prevention of recurrent embolic events are needed.


A single bolus dose of esmolol in the prevention of intubation-induced tachycardia and hypertension in an ambulatory surgery unit.
The efficacy of a single bolus dose of esmolol in the prevention of intubation-induced tachycardia and hypertension was studied in a double-blind manner. Thirty patients from the Ambulatory Surgery Unit at Rush-Presbyterian-St. Luke's Medical Center were prospectively randomized to receive a placebo, 100 mg of esmolol, or 200 mg of esmolol immediately prior to induction (2.5 to 3.0 minutes before intubation). The groups were similar in demographic characteristics and with regard to preoperative blood pressure (BP) and heart rate (HR). Anesthetic management was standardized for all patients. Esmolol 100 mg (1.4 +/- 0.3 mg/kg) and 200 mg (2.6 +/- 0.7 mg/kg) significantly (p less than 0.05) blunted the maximum increases in HR and BP following intubation. The average maximum HR increase in the placebo group was 40% as opposed to 16% in the esmolol 100 mg group and 14% in the esmolol 200 mg group. Both esmolol groups blunted the tachycardic response over a 4-minute postintubation time period. The average maximum BP increase was 47% in the placebo group versus 22% and 19% in the esmolol 100 mg and esmolol 200 mg groups, respectively. There were no significant differences between the two esmolol groups. This study demonstrates the efficacy of a single bolus dose of esmolol in blunting the tachycardic and hypertensive responses to laryngoscopy and intubation in an ambulatory surgery setting.


Madurahydroxylactone, an Inhibitor of Staphylococcus aureus FtsZ from Nonomuraea sp. AN100570.
FtsZ, a bacterial cell-division protein, is an attractive antibacterial target. In the screening for an inhibitor of Staphylococcus aureus FtsZ, madurahydroxylactone (1) and its related derivatives 2-5 were isolated from Nonomuraea sp. AN100570. Compound 1 inhibited S. aureus FtsZ with an IC50 of 53.4 μM and showed potent antibacterial activity against S. aureus and MRSA with an MIC of 1 μg/ml, whereas 2-5 were weak or inactive. Importantly, 1 induced cell elongation in the cell division phenotype assay, whereas 2-5 did not. It indicates that 1 exhibits its potent antibacterial activity via inhibition of FtsZ, and the hydroxyl group and hydroxylactone ring of 1 are critical for the activity. Thus, madurahydroxylactone is a new type of inhibitor of FtsZ.


The past, present and future of gene targeting in Drosophila.
The beginnings of Drosophila as a model organism reach far back into the 1900s to Thomas Hunt Morgan's first fly room. The success of this system for the study of genetics is closely linked to the fact that the fly is amenable to complex genetic manipulations so that random mutagenesis screens can be easily performed. Nonetheless, current advances in genomics and in our ability to predict protein function emphasize the importance of mutagenesis methods that are not random, but rather give the researcher control over how the gene is modified. Gene targeting in Drosophila, developed almost a decade ago, makes use of the organism's own DNA repair machinery to exchange genetic information between a chromosomal target and an exogenous template. Here we discuss available targeting methods and recent advances that facilitate repeated targeting and open the doors to routine allelic studies.


Differences in N-glycosylation of recombinant human coagulation factor VII derived from BHK, CHO, and HEK293 cells.
BACKGROUND & Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. rFVII was purified from selected production clones derived from BHK, CHO, and HEK293 cells after stable transfection, and rFVII isolates were analyzed for protein activity, impurities and post-translational modifications. RESULTS & The analytical results showed no apparent gross differences between the various FVII proteins, except in their N-linked glycosylation pattern. Most N-glycans found on rFVII produced in HEK293 cells were not detected on rFVII from CHO and BHK cells, or, somewhat unexpectedly, on pdFVII; all other protein features were similar. HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). All HEK293rFVII oligosaccharides contained one or more fucoses (Fuc), as well as hybrid and high mannose (Man) structures. From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells were therefore assumed to be the best option for the production of rFVII.


Immunotoxicologic effects of cyclosporine on tumor progression in models of squamous cell carcinoma and B-cell lymphoma in C3H mice.
Many immunosuppressive drugs are associated with an increased risk of neoplasia, principally non-melanoma skin cancers and B-cell lymphomas. However, only 6 of the 13 immunosuppressive drugs tested in 2 year bioassays increased the incidence of neoplasia. For example, the 2-year bioassays conducted with cyclosporine (CSA), an International Agency for Research on Cancer (IARC) Group 1 human carcinogen, were negative. The purpose of these investigations was to use transplanted tumor models in immunocompetent, syngeneic mice to gain insight into the failure of the 2-year bioassay to show an increased incidence of neoplasia with CSA. C3H HeN mice were used in a battery of assays with a transplanted squamous cell carcinoma (SCC VII cells) or a B-cell, lymphoma (38C13 cells) cells to study effects of CSA on local growth and metastases, experimental metastases, and progression of established metastases. Mice received CSA twice weekly by subcutaneous (SC) injection at doses of 0.5, 5, or 50 mg/kg; controls received the CSA vehicle. CSA had a modest inhibitory effect on SC tumors initiated by 38C13 cells and on intramuscular tumors initiated by SCC VII cells. CSA also decreased the number of lung colonies and decreased the size, growth fraction and vascularity of established lung metastases initiated by SCC VII cells. In contrast, CSA increased progressive growth of metastases to the sentinel lymph node from an intramuscular SCC VII tumor, but had no effect cellular traffic to the node. In conclusion, CSA at doses up to 50 mg/kg did not facilitate tumor progression and it partially inhibited tumor growth, suggesting that suppression of tumor progression may partially explain the failure of CSA to act as a carcinogen in 2 year bioassays.


Association between adherence to physical activity guidelines and health-related quality of life among individuals with physician-diagnosed arthritis.
To examine the association between adherence to physical activity guidelines and health-related quality of life (HRQOL) among individuals with arthritis. A cross-sectional sample with 33,071 US adults, 45 years or older with physician-diagnosed arthritis was obtained from 2007 Behavioral Risk Factor Surveillance System survey. We conducted negative binomial regression analysis to examine HRQOL as a function of adherence to physical activity guidelines controlling for physicians' recommendations for physical activity, age, sex, race, education, marital status, employment, annual income, health insurance, personal physician, emotional support, body mass index, activity limitations, health status, and co-morbidities based on Behavioral Model of Health Services Utilization. Descriptive statistics showed that 60% adults with arthritis did not adhere to physical activity guidelines, mean physically and mentally unhealthy days were 7.7 and 4.4 days, respectively. Results from negative binomial regression indicated that individuals who did not adhere to physical activity guidelines had 1.14 days more physically unhealthy days and 1.12 days more mentally unhealthy days than those who adhered controlling for covariates. Adherence to physical activity is important to improve HRQOL for individuals with arthritis. However, adherence is low among this population. Interventions are required to engage individuals with arthritis in physical activity.


Clinical impact of the troponin 99th percentile cut-off and clinical utility of myoglobin measurement in the early management of chest pain patients admitted to the Emergency Cardiology Department.
To verify the clinical impact of different low cut-offs for troponin I/cardiac troponin I (99th percentile to 10% CV) and for myoglobin, in early risk stratification of patients with suspected acute coronary syndrome. A total of 516 consecutive non-ST-elevation patients admitted to hospital were followed. The first measurement of cardiac markers was performed at the point-of-care in the Emergency Cardiology Department, using Stratus CS. The lowest cardiac troponin I concentration with a CV<or=10% (cardiac troponin I concentration=0.07 microg/l) was used to perform an early diagnosis of cardiac damage and to admit non-ST-elevation patients to the Intensive Cardiac Unit. Final diagnosis of acute myocardial infarction was assessed according to European Society of Cardiology and American College of Cardiology diagnostic criteria: cardiac marker follow-up after hospital admission was performed in central laboratory. We retrospectively assessed how the diagnostic accuracy of an early diagnosis of myocardiac damage in the same population might have changed if different lower cardiac troponin I cut-offs had been used upon admitting patients in the Emergency Cardiology Department, independently from the analytical imprecision of the method. A diagnosis of acute myocardial infarction was performed on 110 (21.3%) of 516 non-ST-elevation-patients admitted to hospital. Seventy (13.6%) patients had cardiac troponin I >0.07 microg/l in the Emergency Cardiology Department (P>0.05). Using lowering cut-off values, the difference between the fraction of patients that was positive compared with the diagnosis according to European Society of Cardiology and American College of Cardiology criteria and had remained statistically significant (P<0.05) up to 0.03 microg/l (99th percentile upper reference limit) was considered (85 patients, 16.5%, n.s.). Relative operating characteristic analysis confirmed that the best clinical cut-off was related to the cardiac troponin I concentration that meets the 99th percentile upper reference limit. The diagnostic accuracy of myoglobin in detecting the minimum cardiac damage was significantly lower, independently from the cut-offs considered. The diagnostic accuracy in detecting myocardial damage early in the Emergency Cardiology Department improves when the 99th percentile is used as a decisional value of cardiac troponin I; the use of this cut-off makes the measurement of myoglobin unnecessary.


COVID-19 Lockdown in Patients with Chronic Diseases: A Cross-Sectional Study.
Background: We aimed to investigate the impact of the first COVID-19 lockdown on medication adherence, physician access, lifestyle behaviours, and mental health in patients with chronic conditions. Methods: A cross-sectional phone survey was conducted in 1274 housebound adults recruited from 8 regional chronic disease cohorts (CLEO CD study: NCT04390126). Results: Medication adherence was 97%; 305 (41%) patients declared that at least one scheduled visit with a physician was missed during the first lockdown. The main changes in lifestyle behaviours were deterioration in sleep time (duration and/or quality; 71%), increase in screen time (46%), and decrease in physical activity (46%). Nineteen percent experienced psychological distress (Kessler-6 score ≥ 5). An urban living place (OR, 1.76 vs. rural; 95% CI, 1.32−2.33; p = 10−4), worse self-reported mental health (OR, 1.62 vs. about the same or better; 95% CI, 1.17−2.25; p = 0.003), and a K6 score ≥ 5 (OR, 1.52 vs. <5; 95% CI, 1.05−2.21; p = 0.03) were independent factors associated with at least one unhealthy behaviour. Conclusions: Encouraging results were observed in terms of medication adherence. Caution is needed in chronic disease patients living in urban places as well as those presenting psychological distress and worse self-reported mental health to reduce unhealthy behaviours.


Combing oncolytic adenovirus expressing Beclin-1 with chemotherapy agent doxorubicin synergistically enhances cytotoxicity in human CML cells in vitro.
Cancer virotherapy provides a new strategy to treat cancer that can directly kill cancer cells by oncolysis. Insertion of therapeutic genes into the genome of a modified adenovirus, thereby creating a so-called gene-virotherapy that shares the advantages of gene therapy and virotherapy. In this study we investigated whether a strategy that combines the oncolytic effects of an adenoviral vector with the simultaneous expression of the autophagy gene Beclin-1 offered a therapeutic advantage for chronic myeloid leukemia (CML) cells with resistance to chemotherapy and evaluated the synergistic effects of SG511-BECN and doxorubicin (Dox) in human CML cells in vitro. Oncolytic virus SG511-BECN was constructed through introducing the Beclin-1 gene into the oncolytic adenoviral backbone. SG511-BECN displayed significantly improved antileukemia activity on multidrug-resistant CML cell line K562/A02, which was mediated via induction of autophagic cell death. Furthermore, Dox could synergize with SG511-BECN to kill the CML cells by improving the infectious efficiency of the oncolytic adenovirus without causing significant damage to normal human mononuclear cells. The results demonstrate that targeting the autophagic cell death pathway and combination of a chemotherapy agent with oncolytic adenovirus may be a novel strategy for the treatment of leukemia with chemotherapy resistance.