Ultrasonic studies of proton-transfer reactions at the catalytic site of alpha-chymotrypsin.
Ultrasonic relaxation measurements for alpha-chymotrypsin in phosphate, sulfite and arsenate buffers exhibit a high peak of absorption at neutral pH. The analysis is based on: comparison of the relaxation measurements for the enzyme and for the zymogen and inhibited enzyme; X-ray and neutron diffraction data, and high-resolution NMR data. The ultrasonic relaxation is shown to result mainly from a proton-transfer reaction that involves the histidine at the catalytic site (His-57). The question is raised of whether the enhanced ultrasonic effect observed in the enzyme is indicative of a property that plays a part in the catalytic activity.


[Determination of total serum triiodothyronine in hyperthyroidism: comparison of fluorescence polarization and immunoenzymology].
We evaluated the performances of the Abbott fluorescence polarization assay (FPIA) utilizing the TDx system for human total triiodothyronine (T3) in hyperthyroidism. We compared the results with an immunoenzymometric assay (IEA) (Enzymum Test T3 Boehringer-Mannheim). Greatest attention was focused on the diagnosis of hyperthyroidism because detection of subclinical hyperthyroidism is important. The repeatability of the Abbott fluorescence polarization assay was satisfying (m = 8.07 +/- 0.37 nmol.l-1, CV = 4.59%). The reproducibility was tested with Abbott control sera: m = 4.58 +/- 0.53 nmol.l-1 and CV = 11.5 per cent for level M; m = 7.95 +/- 0.66 nmol.l-1 and CV = 8.23 per cent for level H; m = 2.38 +/- 0.39 nmol.l-1 and CV = 16.5 for level L. The correlation of results of the Abbott assay with those of the Boehringer assay was good for samples from hyperthyroid patients. Values for hyperthyroid and euthyroid subjects were resolved slightly better with the Abbott FPIA than with Boehringer IEA. The Abbott total T3 fluorescence polarization assay may have an additional role to play in monitoring thyroid function in patients under iodine treatment (amiodarone) to eliminate a secondary hyperthyroïdism.


Purification and characterization of the rotenone-insensitive NADH dehydrogenase of mitochondria from Arum maculatum.
The non-ionic detergent lauryl dimethylamine N-oxide (LDAO) has been used to extract the NADH dehydrogenases of Arum maculatum mitochondria. Affinity chromatography on 5'-ADP-Sepharose 4B was used to separate the rotenone-sensitive (complex I) NADH dehydrogenase from the rotenone-insensitive NADH dehydrogenase. An 18-fold purification of the rotenone-insensitive NADH dehydrogenase was achieved. The enzyme is specific for NADH with optimal activity around pH 7.2. The apparent Km for NADH is 28 microM, with dichloroindophenol as acceptor at pH 7.2. The rotenone-insensitive NADH dehydrogenase appears to be a flavoprotein and no iron-sulphur centres were detected by electron spin resonance spectroscopy.


Oncoprotein ZNF322A transcriptionally deregulates alpha-adducin, cyclin D1 and p53 to promote tumor growth and metastasis in lung cancer.
This corrects the article DOI: 10.1038/onc.2015.296.


Schisandrin B attenuates bleomycin-induced pulmonary fibrosis in mice through the wingless/integrase-1 signaling pathway.
Purpose/Aim: Pulmonary fibrosis (PF) is characterized by the progressive and ultimately fatal accumulation of fibroblasts and extracellular matrix in the lung that distorts its architecture and compromises its function.Objective: The present study investigated the potential protective effects of schisandrin B (Sch B) on the Wingless/Integrase-1 (Wnt) signaling pathway in attenuating inflammation and oxidative stress in ICR mice.Methods: Sixty healthy ICR mice were randomly divided into the following groups: control group, bleomycin (BLM) group, Sch B low dose (Sch B-L) group, Sch B medium dose (Sch B-M) group, Sch B high dose (Sch B-H) group, and dexamethasone (DXM) group. The expression of transforming growth factor (TGF)-β1 was examined by ELISA. In addition, the levels of superoxide dismutase (SOD), hydroxyproline (HYP), and the total antioxidant capacity (T-AOC) were determined. The protein and mRNA levels of matrix metalloproteinase 7 (MMP7) and β-catenin in mice were analyzed by western blot and quantitative real -quantitative time PCR (qRT-PCR), respectively.Results: Lung tissues from the BLM group exhibited significantly more inflammatory changes and a significantly greater number of collagen fibers than lung tissues from the control group. In addition, the lung tissues from these BLM-treated mice exhibited slightly increased MMP7 and β-catenin protein expression. Lung tissues from the Sch B-H group exhibited fewer inflammatory changes and fewer collagen fibers than lung tissues from the BLM group. Furthermore, the lung tissues from the Sch B-H mice exhibited decreased HYP and TGF-β1 levels, but increased SOD and T-AOC levels.Conclusions: The present study provided evidence that Sch B may be a potential therapeutic agent for the treatment of PF.


The sexually dimorphic expression of L7/SPA, an estrogen receptor coactivator, in zebra finch telencephalon.
Sex differences in the zebra finch (Taeniopygia guttata) brain are robust and include differences in morphology (song control nuclei in males are significantly larger) and behavior (only males sing courtship songs). In zebra finches, hormonal manipulations during development fail to reverse sex differences in song nuclei size and suggest that the classical model of sexual differentiation is incomplete for birds. Coactivators act to initiate transcriptional activity of steroid receptors, and may help explain why hormonal manipulations alone are not sufficient to demasculinize the male zebra finch brain. The present study investigated the expression and localization of L7/SPA (an estrogen receptor coactivator) mRNA and protein expression across the development of zebra finch song nuclei from males and females collected on P1 (song nuclei not yet formed), P10 (posthatch day 10, song nuclei formed), P30 (30 days posthatch, sexually immature but song nuclei formed and birds learning to sing), and adult birds (older than 65 days and sexually mature). Northern blot analysis showed a significant sex difference in P1 and adult L7/SPA mRNA expression while Western blot analysis also showed enhanced expression in the male brain at all age points. Both in situ hybridization and immunohistochemistry demonstrated that L7/SPA mRNA and protein were located in the song nuclei as well as expressed globally. Elevated coactivator expression may be a possible mechanism controlling the development of male song control nuclei, and coactivators such as L7/SPA may be important regulators of the masculinizing effects of estradiol on brain sexual differentiation.


Family 3 beta-glucosidase from cellulose-degrading culture of the white-rot fungus Phanerochaete chrysosporium is a glucan 1,3-beta-glucosidase.
The substrate specificity of an extracellular beta-glucosidase (BGL) from cellulose-degrading culture of the white-rot fungus Phanerochaete chrysosporium was investigated, using a variety of compounds with beta-glucosidic linkages. Amino acid sequencing data for the purified BGL showed that the enzyme is identical to the glycoside hydrolase (GH) family 3 BGL of the same fungus previously reported [Li, B. and Renganathan, V, Appl. Environ. Microbiol., 64, 2748-2754 (1998)]. The BGL can hydrolyze both cellobiose and cellobionolactone, but cellobionolactone was hydrolyzed very much more slowly than cellobiose. Moreover, cellobionolactone inhibited cellobiose hydrolysis by the BGL, suggesting that this enzyme cannot cooperate with cellobiose dehydrogenase (CDH) in cellulose degradation by P. chrysosporium. In addition to cellobiose, BGL utilized various glucosyl-beta-glucosides, such as sophorose, laminaribiose and gentiobiose, as substrates. Among the four substrates, laminaribiose (beta-1,3-glucosidic linkage) was hydrolyzed most effectively. Moreover, the hydrolytic rate of laminarioligosaccharides increased proportionally to the degree of polymerization (DP), and the activity of BGL even towards laminarin with an average DP of 25 was similar to that towards laminaripentaose (DP 5). Therefore, we conclude that the extracellular BGL from P. chrysosporium is primarily a glucan 1,3-beta-glucosidase (EC 184.108.40.206), which might play a role on fungal cell wall metabolism, rather than a beta-glucosidase (EC 18.104.22.168), which might be involved in the hydrolysis of beta-1,4-glucosidic compounds during cellulose degradation.


Polymeric microneedle-mediated transdermal delivery of melittin for rheumatoid arthritis treatment.
Transdermal drug delivery systems for rheumatoid arthritis (RA) have been receiving increasing attention as they can potentially overcome drawbacks which exist in traditional oral or injection strategies, including low patient compliance and serious gastrointestinal side effects. However, transdermal delivery of RA drugs especially biological drugs suffers from low drug delivery efficiency due to the robust skin barrier. Herein, we fabricated melittin-loaded hyaluronic acid (HA) microneedles and investigated their capacity for inhibiting RA. We showed that melittin-loaded HA microneedles possessed high mechanical strength for successful delivery of melittin into the skin and effectively inhibited RA progression in adjuvant induced both rodent and murine models, as shown by results in histological, paw swelling and arthritis score. Furthermore, after modifying HA with cross-linkable groups, the fabricated microneedles with sustained release properties could further improve the therapeutic potency. Cytokine and T cell analysis in the paws and lymphatic organs indicated that the application of microneedles suppressed the levels of pro-inflammation cytokines including IL-17 and TNF-α, and increased the percentage of regulatory CD4 T cells. Our study revealed that polymeric microneedle-mediated transdermal delivery of melittin could serve as a new therapy with high compliance and good therapeutic efficacy for RA and other autoimmune diseases.


C1q augments platelet activation in response to aggregated Ig.
Immune complexes and aggregated IgG (agg-IgG) induce platelet aggregation and the release reaction. Immune complexes also activate the complement system and interact with the complement component C1q. Since platelets possess both Fc and C1q receptors capable of signal transduction, the present study focused on the interaction between these binding sites and platelet activation. Subaggregating doses of agg-IgG (20-400 microg/ml) were identified for washed platelets from each of 11 healthy donors, and platelet aggregation was monitored in the presence or the absence of increasing concentrations of C1q (5-100 microg/ml). C1q produced a dose-dependent potentiation of platelet alphaIIb/beta3 integrin activation, platelet aggregation, and granule secretion when combined with low doses of agg-IgG. C1q alone was without effect. Maximal enhancement of agg-IgG-induced platelet activation was noted at C1q concentrations ranging from 50 to 100 microg/ml. The observed C1q-induced potentiation of platelet aggregation in response to agg-IgG was blocked by polyclonal antibody F(ab')2 directed against platelet binding sites recognizing the collagen-like domain of C1q (cC1qR) or by mAb Fab (IV.3) directed against platelet FcgammaRII receptors. These data suggest a cooperative interaction between platelet FcgammaRII and cC1q receptors and support a potential role for platelet cC1q receptors in pathologic platelet activation by circulating immune complexes often associated with in vivo thrombosis and thrombocytopenia.


αB-Crystallin interacts and attenuates the tyrosine phosphatase activity of Shp2 in cardiomyocytes under mechanical stress.
The small heat shock protein αB-Crystallin (CryAB, HspB5) and SH2 domain-containing tyrosine phosphatase 2 (Shp2) are important molecules in heart response to pathophysiological stress. Here we show that CryAB interacts with and potentially regulates Shp2 catalytic activity in stretched cardiomyocytes. Such an interaction requires CryAB oligomer to attenuate Shp2 activation. Stretched cardiomyocytes show a robust CryAB/Shp2 association accompanied by a reduction in the Shp2 phosphatase activity. Accordingly, CryAB knock-down in cardiomyocytes enhances Shp2 activity induced by mechanical stress. These results revealed a new role for CryAB, as a modulator of Shp2 phosphatase activity during a functionally relevant stimulus in cardiomyocytes.


ACHC, CHAP, or the joint commission: which one is for you?
Selecting the right accreditation organization is an important decision. This month's column reviews the three nationally recognized accreditation organizations, and examines their standards and survey activities. Factors home care organizations should consider when choosing an accreditation body are presented to facilitate knowledgeable decision making.


Impact of beam configuration on VMAT plan quality for Pinnacle3Auto-Planning for head and neck cases.
The purpose of this study was to compare automatically generated VMAT plans to find the superior beam configurations for Pinnacle3 Auto-Planning and share "best practices". VMAT plans for 20 patients with head and neck cancer were generated using Pinnacle3 Auto-Planning Module (Pinnacle3 Version 9.10) with different beam setup parameters. VMAT plans for single (V1) or double arc (V2) and partial or full gantry rotation were optimized. Beam configurations with different collimator positions were defined. Target coverage and sparing of organs at risk were evaluated based on scoring of an evaluation parameter set. Furthermore, dosimetric evaluation was performed based on the composite objective value (COV) and a new cross comparison method was applied using the COVs. The evaluation showed a superior plan quality for double arcs compared to one single arc or two single arcs for all cases. Plan quality was superior if a full gantry rotation was allowed during optimization for unilateral target volumes. A double arc technique with collimator setting of 15° was superior to a double arc with collimator 60° and a two single arcs with collimator setting of 15° and 345°. The evaluation showed that double and full arcs are superior to single and partial arcs in terms of organs at risk sparing even for unilateral target volumes. The collimator position was found as an additional setup parameter, which can further improve the target coverage and sparing of organs at risk.


Catalytic Activity of Silicon Nanowires Decorated with Gold and Copper Nanoparticles Deposited by Pulsed Laser Ablation.
Silicon nanowires (SiNWs) decorated by pulsed laser ablation with gold or copper nanoparticles (labeled as AuNPs@SiNWs and CuNPs@SiNWs) were investigated for their catalytic properties. Results demonstrated high catalytic performances in the Caryl-N couplings and subsequent carbonylations for gold and copper catalysts, respectively, that have no precedents in the literature. The excellent activity, attested by the very high turn over number (TON) values, was due both to the uniform coverage along the NW length and to the absence of the chemical shell surrounding the metal nanoparticles (MeNPs). A high recyclability was also observed and can be ascribed to the strong covalent interaction at the Me-Si interface by virtue of metal "silicides" formation.


Inhibition of autoimmune Chagas-like heart disease by bone marrow transplantation.
Infection with the protozoan Trypanosoma cruzi manifests in mammals as Chagas heart disease. The treatment available for chagasic cardiomyopathy is unsatisfactory. To study the disease pathology and its inhibition, we employed a syngeneic chicken model refractory to T. cruzi in which chickens hatched from T. cruzi inoculated eggs retained parasite kDNA (1.4 kb) minicircles. Southern blotting with EcoRI genomic DNA digests revealed main 18 and 20 kb bands by hybridization with a radiolabeled minicircle sequence. Breeding these chickens generated kDNA-mutated F1, F2, and F3 progeny. A targeted-primer TAIL-PCR (tpTAIL-PCR) technique was employed to detect the kDNA integrations. Histocompatible reporter heart grafts were used to detect ongoing inflammatory cardiomyopathy in kDNA-mutated chickens. Fluorochromes were used to label bone marrow CD3+, CD28+, and CD45+ precursors of the thymus-dependent CD8α+ and CD8β+ effector cells that expressed TCRγδ, vβ1 and vβ2 receptors, which infiltrated the adult hearts and the reporter heart grafts. Genome modifications in kDNA-mutated chickens can be associated with disruption of immune tolerance to compatible heart grafts and with rejection of the adult host's heart and reporter graft, as well as tissue destruction by effector lymphocytes. Autoimmune heart rejection was largely observed in chickens with kDNA mutations in retrotransposons and in coding genes with roles in cell structure, metabolism, growth, and differentiation. Moreover, killing the sick kDNA-mutated bone marrow cells with cytostatic and anti-folate drugs and transplanting healthy marrow cells inhibited heart rejection. We report here for the first time that healthy bone marrow cells inhibited heart pathology in kDNA+ chickens and thus prevented the genetically driven clinical manifestations of the disease.


Thermodynamics and dynamics of two-dimensional systems with dipolelike repulsive interactions.
Thermodynamics and dynamics of a classical two-dimensional system with dipolelike isotropic repulsive interactions are studied systematically using extensive molecular dynamics (MD) simulations supplemented by appropriate theoretical approximations. This interaction potential, which decays as an inverse cube of the interparticle distance, belongs to the class of very soft long-ranged interactions. As a result, the investigated system exhibits certain universal properties that are also shared by other related soft-interacting particle systems (like, for instance, the one-component plasma and weakly screened Coulomb systems). These universalities are explored in this article to construct a simple and reliable description of the system thermodynamics. In particular, Helmholtz free energies of the fluid and solid phases are derived, from which the location of the fluid-solid coexistence is determined. The quasicrystalline approximation is applied to the description of collective modes in dipole fluids. Its simplification, previously validated on strongly coupled plasma fluids, is used to derive explicit analytic dispersion relations for the longitudinal and transverse wave modes, which compare satisfactory with those obtained from direct MD simulations in the long-wavelength regime. Sound velocities of the dipole fluids and solids are derived and analyzed.


Spontaneous pneumothorax in Wegener granulomatosis.
Spontaneous pneumothorax in Wegener granulomatosis (WG) is uncommon. Three cases of pneumothorax that occurred early in the course of this vasculitis are reported. In the first patient, the disorder was disclosed by a pyopneumothorax. In the second patient, a rupture of the subpleural cavitary nodule into the pleural space was observed. In the third patient, the pneumothorax was discovered at the same time as a pulmonary hemorrhage. The three patients improved with immunosuppressive therapy. In WG, the pulmonary nodules are predominantly in the subpleural location, which entails the risk of pneumothorax and therefore require particular attention.


Costs of Physician-Hospital Integration.
Given that the enactment of the Patient Protection and Affordable Care Act of 2010 is expected to generate forces toward physician-hospital integration, this study examined an understudied, albeit important, area of costs incurred in physician-hospital integration. Such costs were analyzed through 24 semi-structured interviews with physicians and hospital administrators in a multiple-case, inductive study. Two extreme types of physician-hospital arrangements were examined: an employed model (ie, integrated salary model, a group of physicians integrated by a hospital system) and a private practice (ie, a physician or group of physicians who are independent of economic or policy control). Interviews noted that integration leads to 3 evident costs, namely, monitoring, coordination, and cooperation costs. Improving our understanding of the kinds of costs that are incurred after physician-hospital integration will help hospitals and physicians to avoid common failures after integration.


[The status of extrapulmonary tuberculosis in community hospital].
Fifty patients with extrapulmonary tuberculosis were diagnosed at 4 community hospitals in Tokyo during 1981 to 1992. The percentage of extrapulmonary tuberculosis among the all types of tuberculosis was 10.5% as a whole, and 16% in females and 7.9% in males. The number of patients older than fifth generation was about two times more than that of younger generations. And the patients were more in males than in females under the forties, but were more in females than in males over the sixties. On chest radiogram, there was no pathological findings in 52% of the patients and active pulmonary tuberculosis was detected in 32% of the patients. The majority of the extrapulmonary tuberculosis (30 cases, 60%) was of lymph nodes, especially of cervical lymph nodes, and the remaining were 10 cases of tuberculosis in bone and joint, 4 each in intestine and pericardium, 2 each in liver, peritoneum, kidney, urinary bladder, epididymis and skin. Among 30 cases with tuberculous lymphadenopathy, the disease were observed in cervical lymph nodes by 19 cases, in pulmonary hilar and axillary node by each 4 cases and in abdominal cavity by 3 cases.


15-Deoxy-Delta12,14-prostaglandin J2 regulates endogenous Cot MAPK kinase kinase 1 activity induced by lipopolysaccharide.
Cot is a MAPK kinase kinase that has been implicated in cellular activation and proliferation. Here, we show that the addition of lipopolysaccharide (LPS) to RAW264 macrophages induces a 10-fold increase of endogenous Cot activity, measured as MAPK kinase kinase 1 activity. Taxol, but not phorbol 12-myristate 13-acetate (PMA), induces a similar activation of Cot. A tyrosine kinase activity is involved in Cot activation by LPS. 15-Deoxy-Delta12,14-prostaglandin J2, but not rosiglitazone, blocks Cot activation by LPS. Furthermore, 15-deoxy-Delta12,14-prostaglandin J2 also inhibited the LPS-induced Cot in vitro. However, 15-deoxy-Delta12,14-prostaglandin J2 does not inhibit MAPK kinase 1 or ERK1/ERK2 activation/phosphorylation induced by PMA and mediated by c-Raf. Considering these data, we propose that the inhibition of LPS-induced Cot activation is one mechanism by which 15-deoxy-Delta12,14-prostaglandin J2 acts as an anti-inflammatory.


Fragment- and negative image-based screening of phosphodiesterase 10A inhibitors.
A novel virtual screening methodology called fragment- and negative image-based (F-NiB) screening is introduced and tested experimentally using phosphodiesterase 10A (PDE10A) as a case study. Potent PDE10A-specific small-molecule inhibitors are actively sought after for their antipsychotic and neuroprotective effects. The F-NiB combines features from both fragment-based drug discovery and negative image-based (NIB) screening methodologies to facilitate rational drug discovery. The selected structural parts of protein-bound ligand(s) are seamlessly combined with the negative image of the target's ligand-binding cavity. This cavity- and fragment-based hybrid model, namely its shape and electrostatics, is used directly in the rigid docking of ab initio generated ligand 3D conformers. In total, 14 compounds were acquired using the F-NiB methodology, 3D quantitative structure-activity relationship modeling, and pharmacophore modeling. Three of the small molecules inhibited PDE10A at ~27 to ~67 μM range in a radiometric assay. In a larger context, the study shows that the F-NiB provides a flexible way to incorporate small-molecule fragments into the drug discovery.


Mobilization of peripheral blood stem cells (PBSCs) after etoposide, adriamycin and cisplatin therapy, and a multimodal cell therapy approach with PBSCs in advanced gastric cancer.
The EAP combination of etoposide (ETP), doxorubicin (ADM) and cisplatin (CDDP) has been reported to be highly active for advanced gastric cancer. However, it is associated with severe myelotoxicity, and its use has declined. We examined whether peripheral blood stem cells (PBSCs) could be mobilized during hematopoietic recovery after EAP, and assessed the possibility of using multimodal cell therapy with PBSCs for the treatment of advanced gastric cancer. Five men with advanced gastric adenocarcinoma were enrolled. All patients were chemotherapy-naïve. EAP (ETP, 360 mg/m2; ADM, 40 mg/m2; CDDP, 80 mg/m2) was given to each patient, and myelotoxicity was carefully monitored. Granulocyte colony-stimulating factor was administered after the neutrophil nadir, and PBSCs were collected by leukapheresis during hematopoietic recovery. The median nadir of the neutrophil count after EAP was 225/ml, occurring between day 17 and 20. Sufficient numbers of PBSCs [CD34(+) cells, CFU-GM] could be mobilized in 4/5 patients. A 45-year-old patient with extended lymph node metastasis received high-dose EAP with peripheral blood stem cell transplantation (PBSCT), followed by cancer vaccine therapy with dendritic cells (DCs), induced from cryopreserved PBSCs. Both high-dose EAP with PBSCT and DC-based immunotherapy was safely performed for the first time against gastric cancer. Although associated with severe myelotoxicity, EAP can mobilize sufficient numbers of PBSCs during hematopoietic recovery. Multimodal cell therapy combining high-dose chemotherapy with PBSCT and DC-based immunotherapy is feasible and can be a reasonable approach in advanced gastric cancer.


Comparison of Fused-Ring Electron Acceptors with One- and Multidimensional Conformations.
Three fused-ring electron acceptors (FXIC-1, FXIC-2, and FXIC-3) were designed and synthesized. This FXIC series has similar electron-rich central units and the same electron-poor termini. Due to the different steric structures of fluorene, bifluorenylidene, and spirobifluorene, FXIC-1 is a one-dimensional (1D) crystal, while FXIC-2 and FXIC-3 are multidimensional (MD) amorphous materials. The conformations of the FXIC series have a slight impact on their absorption and energy levels. FXIC-1 has higher electron mobility than FXIC-2 and FXIC-3. When blending with different polymer donors (PTB7-Th, J71, and PM7), the FXIC-1-based organic solar cells have efficiencies higher than those of the FXIC-2/FXIC-3-based cells. Meanwhile, the ternary-blend cells based on PTB7-Th:F8IC with FXIC-1, FXIC-2, and FXIC-3 show similar efficiencies, which are all better than those of the binary-blend devices.


Polyion oligonucleotide-decorated gold nanoparticles with tunable surface charge density for amplified signal output of potentiometric immunosensor.
Methods based on nanostructures have been developed for potentiometric immunosensors, but most involve low sensitivity or weak signal output and are unsuitable for routine use in diagnosis. Herein, we devise an in-situ signal-amplification strategy for enhanced electrical readout of potentiometric immunosensor toward target prostate-specific antigen (PSA, one kind of cancer biomarkers), based on polyion oligonucleotide-labeled gold nanoparticles (AuNPs). To decrease the background signal, monoclonal anti-human PSA capture antibody was covalently conjugated onto an activated glassy carbon electrode via typical carbodiimide coupling. AuNPs heavily functionalized with the polyion oligonucleotides and polyclonal anti-PSA detection antibodies (pAb2-AuNP-DNA) were utilized as the signal-generation nanotags. In the presence of target PSA, a sandwich-type immunoreaction was executed between capture antibody and detection antibody on the electrode. The detectable signal derived from the shift in the electric potential as a result of the change in the surface charge before and after the antigen-antibody reaction. With target PSA increased, the captured pAb2-AuNP-DNA to the electrode accompanying detection antibody increased, thereby resulting in the change of the electrode potential. Due to numerous polyion oligonucleotides with the negative charge, the signal readout amplified. Under the optimal conditions, the shift in the output potential was proportional to the logarithm of target PSA concentration and displayed a dynamic linear range from 0.05 to 20 ng mL-1 with a detection limit of 13.6 pg mL-1. An intermediate precision of ≤13.2% was accomplished with the batch-to-batch identification. The selectivity was acceptable. The method accuracy was evaluated for human serum specimens, and gave the consistent results between the potentiometric immunosensor and the referenced enzyme-linked immunosorbent assay (ELISA).


Novel immunological approaches for the treatment of Alzheimer's disease.
Alzheimer's disease (AD), the most prevalent form of dementia worldwide, can be deemed as the next global health epidemic. The biochemistry underlying deposition of amyloid beta (A β) and hyperphosphorylated tau aggregates in AD has been extensively studied. The oligomeric forms of A β that are derived from the normal soluble A β peptides are believed to be the most toxic. However, it is the fibrillar Aβ form that aggregates as amyloid plaques and cerebral amyloid angiopathy, which serve as pathological hallmarks of AD. Moreover, deposits of abnormally phosphorylated tau that form soluble toxic oligomers and then accumulate as neurofibrillary tangles are an essential part of AD pathology. Currently, many strategies are being tested that either inhibit, eradicate or prevent the development of plaques in AD. An exciting new approach on the horizon is the immunization approach. Dramatic results from AD animal models have shown promise for active and passive immune therapies targeting A β. However, there is very limited data in humans that suggests a clear benefit. Some hurdles faced with these studies arise from complications noted with therapy. Encephalitis has been reported in trials of active immunization and vasogenic edema or amyloid - related imaging abnormalities (ARIA) has been reported with passive immunization in a minority of patients. As yet, therapies targeting only tau are still limited to mouse models with few studies targeting both pathologies. As the majority of approaches tried so far are based on targeting a self - protein, though in an abnormal conformation, benefits of therapy need to be balanced against the possible risks of stimulating excessive toxic inflammation. For better efficacy, future strategies will need to focus on the toxic oligomers and targeting all aspects of AD pathology.


Clinical trials of endosseous implants: issues in analysis and interpretation.
The majority of contemporary endosseous dental implant systems are based on designs and materials that, over the last three decades, have proved to be predictably reliable. With proper surgical and prosthetic protocols, rates of implant loss have been held to 15% or less over a 5-year period. This information was obtained largely through longitudinal descriptive studies, primarily aimed at obtaining implant survival rates under ideal clinical conditions, with strict inclusion and exclusion criteria for admitting patients into the studies. It is important to emphasize that under conditions of routine clinical practice, where patient selection may be more relaxed than in clinical trials and clinicians attempt to stretch the limits of current technology, the survival rates may not necessarily match those reported in the literature. Since "surviving" implants may exhibit characteristics likely to lead to eventual loss of the implant, for example severe osseous defects, such implants may not necessarily be considered successful. Successful implants should fulfill a list of other criteria considered essential for long-term survival. Differences in implant design preclude some of these criteria from being uniformly applied to all systems. There is a need to identify criteria for success that can be applied to the majority of implant systems. Implants that fail to meet these criteria should be considered failures. Since failure rates may include "failed" as well as "failing" ("ailing") implants, the two categories should be listed separately. From a practical standpoint, implant failures can be grouped into "early" failures, primarily the result of surgical and/or postoperative complications, and "late" failures that arise during and following the restorative phase. The ability of individual systems to achieve excellent success rates, despite some major differences in their design from other systems, suggests that some requirements, initially considered essential for success, may not be as critical as originally believed. Examples include the need for submerging implants during initial wound healing or the need for stress breaking devices. On the other hand, a basic requirement for implant success, such as primary stability at the time of insertion and following loading of the implant, may be the unifying principle behind the need for adequate bone volume and density, longer or wider implants, and the 3 to 6-month delay recommended before implants are placed in function. With relatively low failure rates, a large number of patients may have to be included in long-term clinical trials before a statistically significant association can be established between failure rates and potential contributing factors. For the same reasons, and to avoid type 2 errors, large populations may be needed to show that two systems have comparable success rates. Proving the superiority of one system over another may require fewer subjects. Given the overall low failure rate and the tendency of failures to cluster in individual subjects, failure rates could be markedly affected by the attrition of a few critical subjects. Additional research is needed to validate methods in current use for the clinical determination of osseointegration, and the diagnosis and treatment of occlusal trauma and microbial infections around implants. Also, more reliable methods are needed for the identification of the primary cause(s) of implant morbidity; i.e., infection or occlusal factors.


Squamous cell carcinoma of the tongue in a patient with Fanconi's anemia: a case report and review of the literature.
Fanconi's anemia (FA) is an autosomal recessive disorder characterized by constitutional aplastic anemia and congenital abnormalities. Patients with this disorder are prone to develop leukemia. Besides the risk of squamous cell carcinoma (SCC), development especially in the head and neck region is also increased. Up to now 40 patients with FA have been reported to develop SCC, and in 14 of them the tongue was the primary site. All of the reported SCC in FA patients originated in mucosal and mucocutaneous sites, especially oral (n=25) and anogenital sites (n=8) and the esophagus (n=6), with the exception of two patients with multiple cutaneous involvement. We report a new case of SCC of the tongue in a patient with FA and review the previous SCC cases.


Virulence Factor Genes in Staphylococcus aureus Isolated From Diabetic Foot Soft Tissue and Bone Infections.
The aim of this study is to describe the presence of genes encoding for 4 virulence factors (pvl, eta, etb, and tsst), as well as the mecA gene conferring resistance to beta-lactam antibiotics, in patients with diabetes and a staphylococcal foot infection. We have also analyzed whether isolates of Staphylococcus aureus from bone infections have a different profile for these genes compared with those from exclusively soft tissue infections. In this cross-sectional study of a prospectively recruited series of patients admitted to the Diabetic Foot Unit, San Juan de Dios Hospital, San José, Costa Rica with a moderate or severe diabetic foot infection (DFI), we collected samples from infected soft tissue and from bone during debridement. During the study period (June 1, 2014 to May 31, 2016), we treated 379 patients for a DFI. S aureus was isolated from 101 wound samples, of which 43 were polymicrobial infections; we only included the 58 infections that were monomicrobial S aureus for this study. Infections were exclusively soft tissue in 17 patients (29.3%) while 41 (70.7%) had bone involvement (osteomyelitis). The mecA gene was detected in 35 cases (60.3%), pvl gene in 4 cases (6.9%), and tsst gene in 3 (5.2%). We did not detect etA and etB in any of the cases. There were no differences in the profile of S aureus genes encoding for virulence factors (pvl, etA, etB, and tsst) recovered from DFIs between those with just soft tissue compared to those with osteomyelitis. However, we found a significantly higher prevalence of pvl+ strains of S aureus associated with soft tissue compared with bone infections. Furthermore, we observed a significantly longer time to healing among patients infected with mecA+ (methicillin-resistant) S aureus (MRSA).


Multidimensional determinants of the caregiving burden among Chinese male caregivers of older family members in Hong Kong.
The objective of this study was to examine multidimensional determinants of the caregiving burden among Chinese male caregivers of older family members in Hong Kong. Based on a modified stress process model, this study explored how background and contextual factors (demographic characteristics and mental health status of caregivers, caregiving-related factors, and support and services), primary stressors (degree of care dependency and special care needs), and secondary stressors (self-efficacy and gender role conflict) might affect the caregiving burden among male caregivers. A questionnaire survey was completed by 204 male caregivers who considered themselves as primary caregivers for older family members with care needs aged 60 and over. Descriptive, bivariate, and multivariate analyses were conducted. Being the son of the care receiver, the presence of depressive symptoms, a higher degree of gender role conflict, a lower degree of caregiving self-efficacy, and the use of support and services all predicted greater caregiving burden among male caregivers. This study established a modified stress process model specifically for male caregivers. Gender role conflict and caregiving self-efficacy were introduced into the model as secondary stressors and found to be significantly associated with the degree of caregiving burden. The findings of this study could inform the development of services and interventions to reduce the caregiving burden among male caregivers.


[Localization of tumor-associated placental protein in human placental tissue and detection in carcinoma cell lines].
Sections of human placental villi tissue and carcinoma cell lines (SMMC-7721, (MGC-803 and Kato-111; were examined using the indirect immunoperoxidase histological technique to further investigate the localization of the tumor-associated placental protein (PP) in human placental tissue and the expression of PP on carcinoma cell lines. PP was prepared from placenta in the first trimester. Placental tissue sections exposed to PP antisera showed a marked staining of the trophoblast cells, especially in syncytiotrophoblast. The strongly stained area was localized to the cell membrane of syncytiotrophoblast. In contrast, the nucleus and the cytoplasm of syncytiotrophoblast were stained weakly. The specific PP antisera also stained the carcinoma cell lines. Among different carcinoma cell lines, the degree of staining of liver carcinoma cell line (SMMC-7721) was higher than that of gastric carcinoma cell lines (MGC-803 and Kato-111). In the carcinoma cell lines the presence of antigen similar to PP suggests that oncogene (one) occur in human trophoblast cells as well as mice trophoblast cells.


Atelosteogenesis type III: orthopedic management.
Atelosteogenesis type III is a rare autosomal dominant skeletal dysplasia caused by mutations in the synthesis of the protein filamin B (FLNB). The mutation in the gene coding for FLNB causes the osteochondrodysplastic features of this disorder. Clinically, osteochondrodysplasia causes unbalanced skeletal maturation and absent or mostly hypoplastic bones, such as the pelvis, vertebrae, ribs, or long bones. In the literature, an orthopedic management for this disorder has not been well described. We report the case and orthopedic management of a 6-year-old female patient with atelosteogenesis type III after 3 years of follow-up.


The social network among the elderly and its relationship with quality of life.
Attention to the needs of the elderly is a social necessity, and it seems that evaluating the social network and quality of life of the elderly can be useful in a better understanding of their needs. This study was performed to determine the relationship between the social network and the quality of life of the elderly in the city of Bojnoord in Iran. In this cross-sectional study, 201 elderly people aged 60 years or more were selected by continuous and consecutive sampling method in Bojnoord, Iran in 2014. Data were collected using demographic questionnaire, Lubben social network scale and LEIPAD elderly quality of life questionnaire. Data were analyzed by SPSS 16 and using descriptive statistics and independent-samples t-test, ANOVA and Pearson product-moment coefficient. The results showed that 30.3% of elderly people studied, were subjected to a high risk of isolation. The highest mean in social network dimensions was in the family (19.68%), friends (12.01%) and the neighbors (9.90%), respectively. The mean score for quality of life of the elderly was as moderate to high (63.90±13.73), and among the quality of life dimensions, the highest mean was related to the self-care dimension (15.59%), and the lowest mean was related to the sexual functioning dimension (1.53%). The findings suggested a positive and significant relationship between social network and quality of life in the elderly who were studied (p<0.000, r=0.468). This study was a step toward understanding the social network status and quality of life of the elderly. It is necessary to say that health care professionals, especially nurses, display a significant role in the community to help people in this regard.


Spectra of base substitution mutations induced in Escherichia coli by tritiated water and the decay of incorporated tritiated thymidine.
The biological effects of tritiated water and of [6-3H]thymidine or [methyl-3H]thymidine incorporated into DNA were compared with those induced by 60Co gamma rays. The killing efficiencies of tritiated water and the tritium-labeled bases were very similar, between 1.8 and 2.0 in terms of the RBE of 60Co gamma rays when compared with the absorbed dose to the bacterial nucleus. The frequency of His+ revertants induced by the decay of [6-3H]thymidine was 3.5 times higher than that induced by [methyl-3H]thymidine or tritiated water; these revertants were most often the result of A:T leads to G:C transitions. In comparison, the other treatments efficiently induced both transitions and transversions. The mutational spectrum resulting from the decay of tritiated water was also determined in the lacI forward-mutagenesis system of Escherichia coli. Transitions predominated at the low dose (2.5 krad), while both transitions and transversions were recovered after a high dose (18 krad). These results are very similar to those observed with 60Co gamma rays and are consistent with the hypothesis that mutagenesis resulting from the decay of [6-3H]thymidine is the result of a position effect, while mutagenesis resulting from the decay of [methyl-3H]thymidine and tritiated water is due to beta-particle ionization.


[Peculiarities of fungal and bacterial microorganisms persistence in the structure of fungal balls of paranasal sinuses].
To compare the data of pathomorphological and microbiological examination of fungal balls removed at paranasal sinuses endoscopic surgery. A total of 148 samples were obtained from patients histologically diagnosed with fungal balls. Microscopic evaluation of the specimens stained according to the Gram method, Gomori-Grocott method and PAS-reaction method, revealed phase changes in the fungal balls conditioned by their different life cycles: the areas of young, mature and degenerating mycelium were detected. Degeneration lead to detritus formation. Detritus areas contained accumulated bacterial colonies. The cultural study of the fungal balls removed from paranasal sinuses, showed fungi growth in 17.6% of cases, while the pathomorphological study confirmed presence of fungi in every investigated sample. Aerobic and anaerobic microbiota of the sinuses contents was represented by multi-agent bacterial and fungal-bacterial associations. Statistical analysis of contingency between culture-positive rate of fungi and different bacteria detected in clinical samples from fungal balls revealed inhibitory influence of Pseudomonas aeruginosa on fungi growth, which might be a reasons of their low isolation rate. Supposedly in some cases the fungal balls consisted of dead fungi, or the sampling was done in the area of the fungal balls degeneration, which might also account for the low sensibility of the cultural method.


Cardiovascular morbidity in rheumatoid arthritis patients in North Canterbury, New Zealand 1999-2008.
Cardiovascular disease is a substantial contributor to increased morbidity and mortality in rheumatoid arthritis (RA). The aim of this audit was to determine the rate of cardiovascular events in a cohort of newly diagnosed RA patients. The inpatient clinical database from Christchurch Hospital, Christchurch, New Zealand, was searched using the International Classification of Diseases 9th Revision (ICD9) and 10 codes representing RA and cardiovascular disease between 1 January 1999 and 31 December 2008. Notes were reviewed with additional demographic and medication data sought. Outpatient data for RA patients was collated from the Rheumatology Department's letter database. Four hundred and six patients were identified with combined ICD9 or 10 codes for RA and ischemic heart disease, of whom 194 had a confirmed myocardial event. Of these, 34 were diagnosed with RA between January 1999 and December 2008 prior to their myocardial event. Kaplan-Meier analysis showed risk of a cardiovascular event at 1 and 10 years was 0.64% and 9.4%, respectively. There were 26 confirmed deaths in the study period. The risk of death at 1 and 10 years was 0.48% and 8.16%, respectively. We have shown a relatively low prevalence of cardiovascular events in this RA population diagnosed within a 10 year period. This is consistent with other reports and likely reflects the short follow-up period. Prospective longer-term studies will be required to further investigate the relative contribution of disease activity and other parameters to cardiovascular events in patients with early RA.


The use of bedside echocardiography for measuring cardiac index and systemic vascular resistance in pediatric patients with septic shock.
Follow-up of cardiac index and systemic vascular resistance index by bedside echocardiography until resuscitation. A set of hemodynamic parameters was obtained, including cardiac output, stroke volume, cardiac index, systemic vascular resistance index, velocity time integral, myocardial performance index, capillary refill time, and heart rate at 0 hours after fluid boluses before the start of inotropes, and followed up after 6 hours and 24 hours. Included were 45 patients with community-acquired septic shock. Septic foci were gastroenteritis (24%), intestinal perforation requiring emergency surgery (24%), pneumonia (20%), central nervous system infection (22%) and soft tissue infection (8%). Klebsiella and Enterobacter were the most frequent isolates. We estimated the factors affecting the cardiac index: high central venous pressure at zero time (r = 0.33, p = 0.024) and persistently high heart rate at hour 6 (r = 0.33, p = 0.03). The systemic vascular resistance index was high in most patients at 0 and 24 hours and at the time of resuscitation and inversely affected the cardiac index as well as affecting the velocity time integral (r = -0.416, -0.61, 0.55 and -0.295). Prolonged capillary refill time was a clinical predictor of the low velocity time integral at 24 hours (r = -0.4). The mortality was 27%. Lower systemic vascular resistance index and higher cardiac output were observed in nonsurviving patients. There was a persistently high systemic vascular resistance index in cold shock patients that influenced the stroke volume index, cardiac index, and velocity time integral. The use of echocardiograms for hemodynamic measurements is important in pediatric septic shock patients to adjust dilators, and vasopressor doses and achieve resuscitation targets in a timely manner.


Warm Ischemia Time at Vascular Anastomosis is an Independent Predictor for Delayed Graft Function in Kidney Transplant Recipients.
Delayed graft function after kidney transplant can affect patient and graft survival, resulting in prolonged hospital stay and need for dialysis. Ischemia times during organ procurement and reanastomosis at transplant are key factors in delayed graft function. We analyzed all living- and deceased-donor renal transplants in Ireland over a 33-month period, with effect of warm ischemia time during anastomosis on delayed graft function being the primary outcome. We performed statistical regression analyses to account for confounding variables. Patients had identical surgical technique and immunosuppression protocols. Of 481 transplants during the study period, 20 patients were excluded because of paired-kidney exchange, nephron dosing transplant, or simul-taneous pancreas-kidney transplant. In the donor pool, 70% were donors after brainstem death, 3.6% were donors after cardiac death, and 26% were living donors. All living donors were direct altruistic donors and underwent stringent assessment via the ethics committee and multidisciplinary team meeting. Of living donors, 8% were not related. These were true altruistic donors who were acquaintances of the recipients and volunteered themselves for assessment. They were assessed in accordance with the declaration of Istanbul and received no compensation of any kind for donation. Of total patients, 18% had delayed graft function, defined as need for dialysis within 7 days of transplant. Warm ischemia time during anastomosis significantly affected risk of delayed graft function but not graft survival or function at 3 months. This factor did not correlate with hospital stay duration. Time on dialysis and recipient weight significantly correlated with risk of delayed graft function. Our findings support a role for minimizing warm ischemia time during anastomosis to reduce delayed graft function and need for dialysis in the perioperative period. However, a longer time does not appear to affect creatinine levels and therefore graft function at 3 months.


FISH and related techniques in the diagnosis of lymphoma.
Many malignant lymphomas are characterized by recurrent genetic abnormalities. These include numerical abnormalities, deletions and reciprocal translocations. In this chapter, we have focused on the detection of chromosomal translocations in B cell lymphomas and discussed some trisomies in lymphomas and CLL. FISH is a well developed molecular method by which it is possible to detect numerical and structural chromosomal abnormalities. We addressed various aspects of metaphase, and especially interphase, FISH and also described the recently developed DNA fibre FISH technology. Using this method, it is possible simultaneously to detect and map chromosomal breakpoints. FISH is also compared with more conventional detection methods such as banding analysis, Southern blot analysis and PCR for the translocations t(8;14) and variant translocations in Burkitt's lymphoma, t(14;18) in follicular lymphoma and t(11;14) in MCL. Other breakpoints in B cell lymphoma are also discussed. It might be concluded that the rapid development in interphase and DNA fibre FISH will provide us with quick, easy and cheap tools to identify specific chromosomal translocations and other genomic abnormalities in human tumours.


Segmental dilatation of the ileum.
One of the rarer causes of intestinal obstruction in the neonatal period is segmental dilatation of the ileum. The case histories, radiological and histological details of four patients are presented together with a short review of the literature.


Dynamic Graph Theoretical Analysis of Functional Connectivity in Parkinson's Disease: The Importance of Fiedler Value.
Graph theoretical analysis is a powerful tool for quantitatively evaluating brain connectivity networks. Conventionally, brain connectivity is assumed to be temporally stationary, whereas increasing evidence suggests that functional connectivity exhibits temporal variations during dynamic brain activity. Although a number of methods have been developed to estimate time-dependent brain connectivity, there is a paucity of studies examining the utility of brain dynamics for assessing brain disease states. Therefore, this paper aims to assess brain connectivity dynamics in Parkinson's disease (PD) and determine the utility of such dynamic graph measures as potential components to an imaging biomarker. Resting-state functional magnetic resonance imaging data were collected from 29 healthy controls and 69 PD subjects. Time-varying functional connectivity was first estimated using a sliding windowed sparse inverse covariance matrix. Then, a collection of graph measures, including the Fiedler value, were computed and the dynamics of the graph measures were investigated. The results demonstrated that PD subjects had a lower variability in the Fiedler value, modularity, and global efficiency, indicating both abnormal dynamic global integration and local segregation of brain networks in PD. Autoregressive models fitted to the dynamic graph measures suggested that Fiedler value, characteristic path length, global efficiency, and modularity were all less deterministic in PD. With canonical correlation analysis, the altered dynamics of functional connectivity networks, and particularly dynamic Fiedler value, were shown to be related with disease severity and other clinical variables including age. Similarly, Fiedler value was the most important feature for classification. Collectively, our findings demonstrate altered dynamic graph properties, and in particular the Fiedler value, provide an additional dimension upon which to non-invasively and quantitatively assess PD.


Antibodies to avian infectious bronchitis virus in Pakistani chickens.
Using the hemagglutination inhibition test, sera from 20 diseased and 3 apparently healthy chicken flocks in Pakistan were examined for antibodies to four types of avian infectious bronchitis virus (AIBV). These flocks were not vaccinated against AIBV. Of the 900 serum samples from diseased flocks, 78 (8.7%) had antibodies to the Massachusetts (M41) type, 23 (2.6%) to the Arkansas type and 20 (2.2%) to the Connecticut type of AIBV. None had antibodies to the JMK type. None of the sera (n = 100) from apparently healthy layers was positive to any of the four AIBV types tested. Some of the birds with antibody titers exhibited no signs of illness except that they laid pale colored eggs or had a history of poor hatchability. These results indicate that at least three AIBV types are circulating in chicken flocks in Pakistan.


Providing Sexual Companionship for Resources: Development, Validation, and Personality Correlates of the Acceptance of Sugar Relationships in Young Women and Men Scale (ASR-YWMS).
A sugar relationship is a transactional sexual relationship in which a younger partner (sugar baby/boy) offers companionship and sexual services to a much older partner (sugar daddy/mommy) in return for material compensation. One aim of the present study was to develop an attitude scale assessing young women's and men's acceptance of sugar relationships. Another aim was to explore the possible associations of the acceptance of sugar relationships with psychological functioning in an intimate partner relationship and in a sexual relationship and with certain socially undesirable personality traits. Two online studies were conducted with a total number of 2052 participants (1879 women; age = 18-28 years). The results show that the Acceptance of Sugar Relationships in Young Women and Men Scale (ASR-YWMS) is a reliable and valid measure of young people's attitude toward sugar relationships. The studies revealed that young women's and men's accepting attitude toward sugar relationships was positively associated with unrestricted sociosexuality, a game-playing love style (Ludus), self-focused sexual motivation (Study 1; N = 319; 272 women and 47 men), and with socially undesirable traits such as Machiavellianism, subclinical psychopathy, and a borderline personality organization (Study 2; N = 1733; 1607 women and 126 men). These findings suggest that a relatively high level of acceptance of sugar relationships is part of a mating strategy focused on opportunities of maximizing resources. This utilitarian, risk-taking and exploitative attitude is characteristic to a fast life history strategy, and it is a fundamental organizing principle of psychological and sexual functioning in intimate partner relationships.


Dipole-bound CH3CN- ions: temperature dependence of ion production rates and lifetimes.
The formation of long-lived (tau less, similar10 mus) dipole-bound CH(3)CN(-) ions through electron transfer in K(14p)CH(3)CN collisions is investigated as a function of target temperature. The rate for their formation is observed to decrease steadily with increasing target temperature. The results are consistent with earlier suggestions that only target molecules in the ground vibrational state and low-lying rotational states can form long-lived dipole-bound anions. For CH(3)CN, the data indicate that creation of long-lived ions requires that the target molecules be in states with rotational quantum numbers j less, similar20. The measurements further demonstrate that the lifetime of the longest-lived (tau greater, similar50 mus) ions is limited by blackbody-radiation-induced photodetachment.


Analysis of phenolic and other aromatic compounds in honeys by solid-phase microextraction followed by gas chromatography-mass spectrometry.
The solid-phase microextraction (SPME) followed by gas chromatography-mass spectrometry (GC-MS) was used for the analysis of phenolic and other aromatic compounds in honey samples from different floral origin. Different parameters affecting the efficiency of the extraction, such as the type of the stationary phase of the fiber, NaCl and acetic acid addition, and extraction time, were optimized for the detection of the maximum number of compounds in the shortest analysis time. A total of 31 compounds were detected, with most of them identified and quantified by GC-MS. The principal component analysis (PCA) was applied to the data matrix; the results allowed for the differentiation between honeydew and nectar honeys on the basis of the salicylic acid concentration. It was found that this acid has a high contribution in the honeydew group (71.2-705.9 microg/100 g of honey) compared to the nectar honey group (0-47.6 microg/100 g of honey). The comparison of data in each honey group enabled us to characterize the floral source of some honeys using some aromatic compounds as markers.


Citrate synthase proteins in extremophilic organisms: studies within a structure-based model.
We study four citrate synthase homodimeric proteins within a structure-based coarse-grained model. Two of these proteins come from thermophilic bacteria, one from a cryophilic bacterium and one from a mesophilic organism; three are in the closed and two in the open conformations. Even though the proteins belong to the same fold, the model distinguishes the properties of these proteins in a way which is consistent with experiments. For instance, the thermophilic proteins are more stable thermodynamically than their mesophilic and cryophilic homologues, which we observe both in the magnitude of thermal fluctuations near the native state and in the kinetics of thermal unfolding. The level of stability correlates with the average coordination number for amino acid contacts and with the degree of structural compactness. The pattern of positional fluctuations along the sequence in the closed conformation is different than in the open conformation, including within the active site. The modes of correlated and anticorrelated movements of pairs of amino acids forming the active site are very different in the open and closed conformations. Taken together, our results show that the precise location of amino acid contacts in the native structure appears to be a critical element in explaining the similarities and differences in the thermodynamic properties, local flexibility, and collective motions of the different forms of the enzyme.


Are Incidental Minute Pulmonary Nodules Ultimately Determined to Be Metastatic Nodules in Esophageal Cancer Patients?
Esophageal cancer patients may simultaneously have resectable esophageal cancer and undiagnosable incidental minute solid pulmonary nodules. While the latter is rarely metastatic, only a few studies have reported on the outcomes of such nodules after surgery. In this retrospective study, we assessed the incidence of such nodules, the probability that they are ultimately metastatic nodules, and the prognosis of patients after esophagectomy according to the metastatic status of the nodules. Data of 398 patients who underwent esophagectomy for resectable esophageal cancer between January 2012 and December 2016 were collected. We reviewed computed tomography (CT) images from the first visit and searched for incidental minute pulmonary nodules <10 mm in size. We followed the outcomes of these nodules and compared the characteristics of metastatic and nonmetastatic nodules. We also assessed the prognosis of patients whose minute pulmonary nodules were metastatic. Among the patients who underwent esophagectomy, 149 (37.4%) had one or more minute pulmonary nodules, with a total of 285 nodules. Thirteen (4.6%) of these nodules in 12 (8.1%) patients were ultimately diagnosed as being metastatic. Thirteen (8.7%) patients experienced recurrence at a different location from where the nodules were originally identified. Characteristics of the metastatic nodules were not unique in terms of size, SUVmax, or location in the lungs. Two-year and 5-year overall survival rates of patients whose nodules were metastatic were 64.2 and 32.1%, respectively. The rate of minute pulmonary nodules which were ultimately metastatic was 4.6%. Our findings suggest that esophagectomy followed by the identification of minute pulmonary nodules is an acceptable strategy even if the nodules cannot be diagnosed as being metastatic on the first visit CT due to their small size.


Long-term follow-up of trigger point injections for abdominal wall pain.
Abdominal wall pain (AWP) is a common yet often overlooked source of abdominal pain. Trigger point injections (TPI) into the abdominal wall have been tried in the past. Few studies have looked at the long-term outcome from these injections. A retrospective chart review was performed on 110 consecutive patients who received TPI for abdominal pain at the University of Western Ontario, London, Ontario. Outcomes from patients whose pain was due to AWP were determined. AWP was defined as fixed or localized pain and superficial or point tenderness (less than 2.5 cm diameter) or a positive Carnett sign (increased pain with tensing abdomen). The primary outcome was long-term efficacy of TPI. The number of diagnostic tests ordered to exclude AWP and the cost of investigating it were determined. Secondary analyses were done to determine if there were significant predictors of response to TPI. Eighty-nine of 110 patients who received TPI met the criteria for AWP. In those who met the criteria for AWP, the average age was 42 years, 84% were female, and the average length of follow-up was 25 months. The primary outcome shows that, at follow-up, 77% had some or complete relief and 23% had no relief. An average of 4.3 diagnostic tests per patient were ordered to exclude other causes of abdominal pain. Secondary analyses show that meeting the criteria for AWP (P<0.0005), the absence of gastrointestinal symptoms (P<0.025), and an upper abdominal location of pain (P<0.025) were statistically significant predictors of a positive response to TPI. This study demonstrates that TPI, in patients who meet criteria for AWP, are effective over the long term.


Preconception care by the nonobstetrical provider.
Clinicians who provide health care to women during their childbearing years have the opportunity to affect pregnancy outcomes positively through preconception care. The goal of preconception care is to identify medical and social conditions that may put the mother or fetus at risk. Key elements include screening for certain infectious diseases, obtaining genetic history, updating immunizations, providing specific nutritional advice, and optimizing health status. Some women, and their partners, may require additional care, including a prepregnancy consultation with an obstetrician or maternal-fetal medicine specialist. The clinician can also offer advice that may enhance conception and encourage either early prenatal care when a pregnancy is achieved or early evaluation for infertility.


Closed-chest robotic repair of mitral prolapse. Surgical technique and early results.
Robotic mitral repair is generally performed with four intercostal trocars and a minithoracotomy. We describe our technique and results with a totally-thoracoscopic closed chest approach using a 12 mm valveless trocar as "working port", without a minithoracotomy. We compared our results with this technique with a control group of robotic mitral repairs performed earlier with a minithoracotomy. Review of all patients with degenerative mitral valve disease who underwent robotic mitral valve repair surgery since December 2019 (n = 110). Patients with concomitant procedures (n = 8) were excluded. The remaining 102 patients were divided in two groups, depending on the approach used, minithoracotomy (n = 63) and totally thoracoscopic (n = 39). There were no significant differences between groups regarding preoperative characteristics. All procedures were completed robotically as planned, and repair rate was 100%. The minithoracotomy group showed a higher percentage of leaflet resections (17.9% vs. 38.7%; p = 0.03). All surgical times were significatively reduced in the totally thoracoscopic group: Cardiopulmonary bypass (97 vs. 115 min, p = 0.0008), ischemic time (67 vs. 80 min, p = 0.0013) and total surgical time (185 vs. 225 min; p < 0.00001). There were no differences in ICU length of stay (1 day, p = 0.07) but hospital length of stay was shorter in the totally thoracoscopic group (4 days; p = 0.0001). Postoperative complications were similar between groups. MR at discharge was mild or less in all cases. Robotic mitral repair for degenerative disease can be safely performed as a closed-chest procedure, using a 12 mm trocar as "working port" and avoiding the need for a minithoracotomy. This approach does not seem to negatively affect the quality of the procedure by any measure, providing similar excellent clinical outcomes and repair rate. All surgical times were shorter in the closed-chest group.


Conversion efficiency of high-Z backlighter materials.
High-Z backlighter materials are commonly used as x-ray sources for diagnosing laser-driven experiments. In order to properly plan for experiments and analyze the data, it is important to understand both the number and distribution of photons emitted by the x-ray source when it is irradiated by a laser. The conversion efficiency of L-shell and M-shell emitters is not as well understood as K-shell emitters. The conversion efficiency of the former is typically presented in terms of the entire L- or M-shell spectral region. However, for some applications, one may only want to use a subset of this spectral region. Laser conversion efficiency for L-shell and M-shell emitters suitable for high-energy (>3 keV) absorption spectroscopy is presented at multiple laser intensities. The measured conversion efficiency of the materials ranges from 0.2% to 0.6%.


Enantioselective construction of quaternary tetrahydropyridines by palladium-catalyzed vinylborylation of alkenes.
Here, the highly enantioselective construction of 3,3-disubstituted tetrahydropyridines via Pd(0)-catalyzed asymmetric vinylborylation of (Z)-1-iodo-dienes and B2pin2 in the presence of a (S)-p-CF3-BnPHOX ligand is reported. This process is supposed to be initiated by oxidative addition of Pd(0) to vinyl iodide, followed by transmetallation with B2pin2, insertion of the pendant alkene and reductive elimination of alkyl-Pd(ii)-Bpin.


Spectral probabilities and generating functions of tandem mass spectra: a strike against decoy databases.
A key problem in computational proteomics is distinguishing between correct and false peptide identifications. We argue that evaluating the error rates of peptide identifications is not unlike computing generating functions in combinatorics. We show that the generating functions and their derivatives ( spectral energy and spectral probability) represent new features of tandem mass spectra that, similarly to Delta-scores, significantly improve peptide identifications. Furthermore, the spectral probability provides a rigorous solution to the problem of computing statistical significance of spectral identifications. The spectral energy/probability approach improves the sensitivity-specificity tradeoff of existing MS/MS search tools, addresses the notoriously difficult problem of "one-hit-wonders" in mass spectrometry, and often eliminates the need for decoy database searches. We therefore argue that the generating function approach has the potential to increase the number of peptide identifications in MS/MS searches.


The role and mechanisms of Microglia in Neuromyelitis Optica Spectrum Disorders.
Neuromyelitis optica spectrum disorder (NMOSD) is an autoimmune neurological disease that can cause blindness and disability. As the major mediators in the central nervous system, microglia plays key roles in immunological regulation in neuroinflammatory diseases, including NMOSD. Microglia can be activated by interleukin (IL)-6 and type I interferons (IFN-Is) during NMOSD, leading to signal transducer and activator of transcription (STAT) activation. Moreover, complement C3a secreted from activated astrocytes may induce the secretion of complement C1q, inflammatory cytokines and progranulin (PGRN) by microglia, facilitating injury to microglia, neurons, astrocytes and oligodendrocytes in an autocrine or paracrine manner. These processes involving activated microglia ultimately promote the pathological course of NMOSD. In this review, recent research progress on the roles of microglia in NMOSD pathogenesis is summarized, and the mechanisms of microglial activation and microglial-mediated inflammation, and the potential research prospects associated with microglial activation are also discussed.


A novel sensitive electrochemical method for the detection of ractopamine in meat food via polycitrulline-modified electrode.
In this paper we report the modification with citrulline of the surface of a glassy carbon electrode (GCE) through cyclic voltammetry to prepare a polycitrulline-modified electrode (PCit/GCE). The electrochemical behaviour of ractopamine on this PCit/GCE was then investigated by cyclic voltammetry, to establish a novel electrochemical method for the detection of ractopamine residues in animal tissue. The optimum preparation conditions for the modified electrode were found to include a polymerisation solution pH of 6.8. The cyclic voltammetry required 12 scan cycles at a scan rate of 40 mV/s with a potential range of -1.6 to 2.3 V. The optimum conditions to determine ractopamine were in a solution of pH 6.0 with cyclic voltammetry performed at a scan rate of 60 mV/s and a potential range of 0.4 to 1.0 V. To analyse meat, the tissue sample was extracted with ethanol, filtered, evaporated to dryness and diluted with phosphate buffered saline at pH of 6.0 before being analysed in the electrochemical cell. The limit of detection of the new method for ractopamine was 1.0 ng/g and the correlation coefficient (R) was 0.9991 in the concentration range of 140-5410 ng/g. Ractopamine residue levels in spiked beef and mutton samples were detected by the modified electrode and the recovery was in the range of 96%-101%, giving results consistent with a validated HPLC method. This study provides a new method for detection and quantification of ractopamine residues in animal tissue.


Development of a mechanical testing and loading system for trabecular bone studies for long term culture.
A highly accurate (+/-3%) mechanical loading and measurement system combined with a trabecular bone diffusion culture-loading chamber has been developed, which provides the ability to study trabecular bone (and possibly) cartilage under controlled culture and loading conditions over long periods of time. The loading device has been designed to work in two main modes, either to apply a specific compressive strain to a trabecular bone cylinder or to apply a specific force and measure the resulting deformation. Presently, precisely machined bone cylinders can be loaded at frequencies between 0.1 Hz to 50 Hz and amplitudes over 7,000 microepsilon. The system allows accurate measurement of many mechanical properties of the tissue in real time, including visco-elastic properties. This paper describes the technical components, reproducibility, precision, and the calibration procedures of the loading system. Data on long term culture and mechanical responses to different loading patterns will be published separately.


[Characterization of immunoreactive hCG beta-subunit in cultured fluids of the cell lines derived from gynecologic malignant tumors].
We characterize the hCG beta-like materials in cultured fluids from 12 different cell lines derived from gynecologic malignant tumors (SKG-1, -2, -3a and -3b cervical squamous carcinoma, SNG-M and -2 endometrial adenocarcinoma, SKN uterine sarcoma, RTSG ovarian undifferentiated adenocarcinoma, RMUG ovarian mucinous adenocarcinoma, RKN ovarian sarcoma, RMG ovarian clear cell carcinoma and NJG gestational choriocarcinoma) by three kinds of enzyme immunoassay (EIA) which were specific for whole hCG, free hCG beta and beta-core fragment, respectively. Of eleven nontrophoblastic cell lines, nine secreted hCG beta-like immunoreactive substance. The above-mentioned three EIAs for each fractionated specimens with gel chromatography on Sephadex G-100 revealed that the immunoreactivity in the SKG-2 and RTSG cultured fluids were totally attributable to free hCG beta but neither to whole hCG nor beta-core fragment. On the other hand, the NJG choriocarcinoma cell line secreted both whole hCG and free hCG beta, no beta-core fragment could be detected in the cultured fluid. The present results suggested that ectopically produced hCG beta-like material may represent the free hCG beta molecule, and that the beta-core fragment may not be a cellular secretory product.


Virucidal activity presence in Trichilia glabra leaves.
Different immunomodulatory activities present in Trichilia glabra (TG) leaf extracts have already been described. Particularly, chloroform-methanol extracts were responsible for an in-vivo anti-inflammatory effect. The effect of such extracts on the infectivity of enveloped and naked viruses were investigated. Methanolic fraction extracts were active against herpes simplex virus type 1 (HSV-1) and vesicular stomatitis virus (VSV), while no activity against poliovirus type 3 was observed. VSV was slightly more affected than HSV-1: 2.8 log10 reduction in VSV titer against 2.4 log10 reduction in HSV-1 titer when 0.25 mg/ml F2 fraction was tested and a reduction of 2.7 log10 in VSV virus titer and of 1.5 log10 in HSV-1 virus titer was observed when 0.25 mg/ml F3 fraction was tested. Results obtained in this work suggest a potential pharmaceutical use of TG extract components.


Mortality, causes of death, and health status among methamphetamine users.
This study examines causes of death, years of life lost, and health and drug use characteristics associated with mortality over an 8 to 10 year period in a sample of methamphetamine users who had and had not received substance use disorder treatment (N = 563). Decedents reported initiating their methamphetamine use for different reasons than surviving methamphetamine users, and some of these differences varied by treatment status. Study findings provide additional detail on long-term health and mortality outcomes in a diverse sample of methamphetamine users, which may inform public health strategies targeting the comparable and divergent needs of treated and untreated populations.


Nepeta Dschuparensis Bornm Extract Moderates COX-2 and IL-1β Proteins in a Rat Model of Cerebral Ischemia.
Nepeta dschuparensis Bornm (NP) is used as a medicinal herb in Iran. In traditional medicine, this herb is extensively employed for curing ailments such as cardiovascular diseases. NP has antioxidant and anti-inflammatory properties. This project examined the effects of the NP extract on cyclooxygenase-2 (COX-2) and interleukin-1β (IL-1β) protein levels and its efficacy in neuroprotection in a cerebral ischemia-reperfusion model. Twenty-six male rats were randomly divided into 3 groups: 1) sham (n=6): no middle cerebral artery occlusion (MCAO) procedure, 2) control (n=10): MCAO procedure and treatment with normal saline, and 3) NP extract (n=10): MCAO procedure and treatment with the NP extract (20 mg/kg, i.p.) at the beginning of reperfusion. To examine the injury caused by cerebral ischemia, we measured motor coordination and the infarct area using the rotarod test and triphenyl tetrazolium chloride staining, respectively. IL-1β and COX-2 protein levels, as inflammatory markers, were measured by immunoblotting assay. The statistical analyses were performed using SPSS, version 16, and the data are expressed as means±SEMs. Statistical difference was evaluated using the one-way ANOVA, followed by the post hoc LSD test (P<0.01). Treatment with the NP extract significantly diminished the infarct volume and alleviated the motor coordination disorder induced by cerebral ischemia. The NP extract administration significantly attenuated the increase in IL-1β and COX-2 protein levels too (P<0.01). The beneficial effects of the NP extract are related to its ability to decrease the levels of IL-1β and COX-2.


Urban area disadvantage and physical activity: a multilevel study in Melbourne, Australia.
To estimate variation between small areas in the levels of walking, cycling, jogging, and swimming and overall physical activity and the importance of area level socioeconomic disadvantage in predicting physical activity participation. All census collector districts (CCDs) in the 20 innermost local government areas in metropolitan Melbourne, Australia, were identified and ranked by the percentage of low income households (<400 dollars/week) living in the CCD. Fifty CCDs were randomly selected from the least, middle, and most disadvantaged septiles of the ranked CCDs and 2349 residents (58.7% participation rate) participated in a cross sectional postal survey about physical activity. Multilevel logistic regression (adjusted for extrabinomial variation) was used to estimate area level variation in walking, cycling, jogging, and swimming and in overall physical activity participation, and the importance of area level socioeconomic disadvantage in predicting physical activity participation. There were significant variations between CCDs in all activities and in overall physical participation in age and sex adjusted models; however, after adjustment for individual SES (income, occupation, education) and area level socioeconomic disadvantage, significant differences remained only for walking (p = 0.004), cycling (p = 0.003), and swimming (p = 0.024). Living in the most socioeconomically disadvantaged areas was associated with a decreased likelihood of jogging and of having overall physical activity levels that were sufficiently active for health; these effects remained after adjustment for individual socioeconomic status (sufficiently active: OR 0.70, 95% CI 0.55 to 0.90 and jogging: OR = 0.69, 95% CI 0.51 to 0.94). These research findings support the need to focus on improving local environments to increase physical activity participation.


Laboratory Evaluation of the Rapid Analyte Measurement Platform Assay to Detect Dengue Virus in Mosquito Pools.
We report the results of a laboratory sensitivity and specificity evaluation of the Rapid Analyte Measurement Platform (RAMP®) Dengue Virus (DENV) antigen detection assay, which is designed to detect all serotypes of DENV in mosquito pools. The RAMP DENV assay was able to detect geographically distinct strains of all 4 DENV serotypes in virus-spiked mosquito pools that contained at least 4.3 log10 plaque forming units/ml, although discrete sensitivity limits varied slightly for each serotype. The RAMP DENV assay also detected DENV 1-4 in mosquito pools containing a single infected mosquito and 24 laboratory-reared uninfected mosquitoes. No false positives were detected in negative control mosquito pools or in samples containing high titers of nontarget arboviruses. We found that while the kit-supplied RAMP buffer reduced the infectious titer of DENV, it did not completely inactivate all serotypes. We recommend adding a detergent, Triton X-100, to the buffer to ensure complete inactivation of DENV if the assay is to be conducted at a lower biosafety level than required for DENV handling.


Characterization of porcine cytokine inducible SH2-containing protein gene and its association with piglet diarrhea traits.
The cytokine inducible SH2-containing protein (CISH), which might play a role in porcine intestine immune responses, was one of the promising candidate genes for piglet anti-disease traits. An experiment was conducted to characterize the porcine CISH (pCISH) gene and to evaluate its genetic effects on pig anti-disease breeding. Both reverse transcription polymerase chain reaction (RT-PCR) and PCR were performed to obtain the sequence of pCISH gene. A pEGFP-C1-CISH vector was constructed and transfected into PK-15 cells to analysis the distribution of pCISH. The sequences of individuals were compared with each other to find the polymorphisms in pCISH gene. The association analysis was performed in Min pigs and Landrace pigs to evaluate the genetic effects on piglet diarrhea traits. In the present research, the coding sequence and genomic sequence of pCISH gene was obtained. Porcine CISH was mainly localized in cytoplasm. TaqI and HaeIII PCR restriction fragment length polymorphism (RFLP) assays were established to detect single nucleotide polymorphisms (SNPs); A-1575G in promoter region and A2497C in Intron1, respectively. Association studies indicated that SNP A-1575G was significantly associated with diarrhea index of Min piglets (p<0.05) and SNP A2497C was significantly associated with the diarrhea trait of both Min pig and Landrace piglets (p<0.05). This study suggested that the pCISH gene might be a novel candidate gene for pig anti-disease traits, and further studies are needed to confirm the results of this preliminary research.


Serum adiponectin levels are inversely associated with overall and central fat distribution but are not directly regulated by acute fasting or leptin administration in humans: cross-sectional and interventional studies.
Adiponectin is an adipocyte-secreted protein that circulates in high concentrations in the serum and acts to increase insulin sensitivity. Previous studies have shown that serum adiponectin is inversely associated with fat mass and insulin resistance in humans and that acute fasting decreases adipose tissue adiponectin mRNA expression in rodents. Whether acute energy deprivation, body fat distribution, or serum hormone levels are associated with circulating adiponectin in humans remains largely unknown. To identify predictors of serum adiponectin levels, we evaluated the association of adiponectin with several anthropometric, metabolic, and hormonal variables in a cross-sectional study of 121 women without a known history of diabetes. We also performed interventional studies to assess whether fasting for 48 h and/or leptin administration regulates serum adiponectin in healthy men and women. Our cross-sectional study shows that, in addition to overall obesity, central fat distribution is an independent negative predictor of serum adiponectin and suggests that adiponectin may represent a link between central obesity and insulin resistance. In addition, estradiol is negatively and independently associated with adiponectin, whereas there is no association between serum adiponectin and leptin, cortisol, or free testosterone levels. Our interventional studies demonstrate that neither fasting for 48 h, resulting in a low leptin state, nor leptin administration at physiological or pharmacological doses alters serum adiponectin levels. Further studies are needed to fully elucidate the physiology of adiponectin in humans and its role in the pathogenesis of insulin-resistant states.


Dissipative dynamics of enzymes.
We explore enzyme conformational dynamics at sub-Å resolution, specifically, temperature effects. The ensemble-averaged mechanical response of the folded enzyme is viscoelastic in the whole temperature range between the warm and cold denaturation transitions. The dissipation parameter γ of the viscoelastic description decreases by a factor of 2 as the temperature is raised from 10 to 45 °C; the elastic parameter K shows a similar decrease. Thus, when probed dynamically, the enzyme softens for increasing temperature. Equilibrium mechanical experiments with the DNA spring (and a different enzyme) also show, qualitatively, a small softening for increasing temperature.


Functional and morphological changes of the gut barrier during the restitution process after hemorrhagic shock.
To investigate the functional, morphological changes of the gut barrier during the restitution process after hemorrhagic shock, and the regional differences of the large intestine and small intestine in response to ischemia/reperfusion injury. Forty-seven Sprague-Dawley rats with body weight of 250-300 g were divided into two groups: control group (sham shock n = 5) and experimental group (n = 42). Experimental group was further divided into six groups (n = 7 each) according to different time points after the hemorrhagic shock, including 0(th) h group, 1st h group, 3rd h group, 6th h group, 12th h group and 24th h group. All the rats were gavaged with 2 mL of suspension of lactulose (L) (100 mg/2 mL) and mannitol (M) (50 mg/each) at the beginning and then an experimental rat model of hemorrhagic shock was set up. The specimens from jejunum, ileum and colon tissues and the blood samples from the portal vein were taken at 0, 1, 3, 6, 12 and 24 h after shock resuscitation, respectively. The morphological changes of the intestinal mucosa, including the histology of intestinal mucosa, the thickness of mucosa, the height of villi, the index of mucosal damage and the numbers of goblet cells, were determined by light microscope and/or electron microscope. The concentrations of the bacterial endotoxin lipopolysaccharides (LPS) from the portal vein blood, which reflected the gut barrier function, were examined by using Limulus test. At the same time point, to evaluate intestinal permeability, all urine was collected and the concentrations of the metabolically inactive markers such as L and M in urine were measured by using GC-9A gas chromatographic instrument. After the hemorrhagic shock, the mucosal epithelial injury was obvious in small intestine even at the 0(th) h, and it became more serious at the 1st and the 3rd h. The tissue restitution was also found after 3 h, though the injury was still serious. Most of the injured mucosal restitution was established after 6 h and completed in 24 h. Two distinct models of cell death-apoptosis and necrosis-were involved in the destruction of rat intestinal epithelial cells. The number of goblet cells on intestinal mucosa was reduced significantly from 0 to 24 h (the number from 243+/-13 to 157+/-9 for ileum, 310+/-19 to 248+/-18 for colon; r = -0.910 and -0.437 respectively, all P<0.001), which was the same with the large intestine, but the grade of injury was lighter with the values of mucosal damage index in 3 h for jejunum, ileum, and colon being 2.8, 2.6, 1.2, respectively. The mucosal thickness and the height of villi in jejunum and ileum diminished in 1 h (the average height decreased from 309+/-24 to 204+/-23 microm and 271+/-31 to 231+/-28 microm, r = -0.758 and -0.659, all P<0.001; the thickness from 547+/-23 to 418+/-28 microm and 483+/-45 to 364+/-35 microm, r = -0.898 and -0.829, all P<0.001), but there was no statistical difference in the colon (F = 0.296, P = 0.934). Compared with control group, the urine L/M ratio and the blood LPS concentration in the experimental groups raised significantly, reaching the peak in 3-6 h (L/M: control vs 3 h vs 6 h was 0.029+/-0.09 vs 0.063+/-0.012 vs 0.078+/-0.021, r = -0.786, P<0.001; LPS: control vs 3 h vs 6 h was 0.09+/-0.021 vs 0.063+/-0.012 vs 0.25+/-0.023, r = -0.623, P<0.001), and it kept increasing in 24 h. The gut barrier of the rats was seriously damaged at the early phase of ischemic reperfusion injury after hemorrhagic shock, which included the injury and atrophy in intestinal mucosa and the increasing of intestinal permeability. Simultaneously, the intestinal mucosa also showed its great repairing potentiality, such as the improvement of the intestinal permeability and the recovery of the morphology at different phases after ischemic reperfusion injury. The restitution of gut barrier function was obviously slower than that of the morphology and there was no direct correlation between them. Compared with the small intestine, the large intestine had stronger potentiality against injury. The reduction of the amount of intestinal goblet cells by injury did not influence the ability of intestinal mucosal restitution at a certain extent and it appeared to be intimately involved in the restitution of the epithelium.


Extremely stable platinum nanoparticles encapsulated in a zirconia nanocage by area-selective atomic layer deposition for the oxygen reduction reaction.
Encapsulation of Pt nanoparticles (NPs) in a zirconia nanocage by area-selective atomic layer deposition (ALD) can significantly enhance both the Pt stability and activity. Such encapsulated Pt NPs show 10 times more stability than commercial Pt/C catalysts and an oxygen reduction reaction (ORR) activity 6.4 times greater than that of Pt/C.


Surveillance of nail gun injuries by journeymen carpenters provides important insight into experiences of apprentices.
Experienced journeymen conducted detailed surveillance interviews with injured apprentice carpenters (n = 413). Nail gun injuries commonly occurred due to inadvertent discharge, ricocheting or projectile nails, and penetration of the wood surface. Framing nailers with contact trip triggers were most often involved; these triggers allow nail discharge anytime the trigger and nose piece are both depressed including following recoil of the tool after firing. Injured workers made concrete and practical suggestions for prevention. Scenarios were identified where changes in work practice, safer triggers, and thoughtful training could prevent injuries. Most injuries were not reported through workers' compensation; this was often due to perceived lack of injury severity, but also involved subtle as well as overt pressures on apprentices not to report. These case-based analyses are consistent with reports that many injuries from nail guns could be prevented with existing safer trigger mechanisms and training. Efforts should also focus on improved injury reporting.


Distal femur reconstruction with modular tumour prostheses: a single Institution analysis of implant survival comparing fixed versus rotating hinge knee prostheses.
To evaluate the incidence of complications in distal femur reconstructions with modular prostheses, comparing fixed vs rotating hinge knee. Retrospective analysis of implant survival, complications, and functional results of the Rizzoli series on distal femur megaprosthesis. Between 1983 and 2010, 687 distal femur tumour modular prostheses were implanted: 491 fixed hinge and 196 rotating hinge knee prostheses. Failures of the prostheses were classified in five types: type 1, soft tissue failure; type 2, aseptic loosening; type 3, structural failure; type 4, infection; type 5, tumour progression. Failure rate was 27 % (185/687). Implant survival to all types of failure was 70 % at ten years and 50 % at 20 years with no significant difference between fixed and rotating hinge knee prostheses (p = 0.0928). When excluding type 5 and type 1 failures, the overall survival was 78 % and 58 % at ten and 20 years. There was not a significant difference in implant survival to aseptic loosening (p = 0.5) and infection (p = 0.2) between fixed and rotating hinge knee prostheses. All cases of breakage of prosthetic components occurred in fixed hinge knee prostheses. Functional results, evaluated in 536 pts, were satisfactory in 91.4 % of cases with a mean score of 23.3 with a significantly better function for rotating hinge knee prostheses (p < 0.001). The most frequent cause of failure was infection followed by aseptic loosening. Even if better results were expected for rotating hinge knee prostheses, there is no significant difference in overall implant survival. No cases of breakage of prosthetic components occurred in rotating hinge knee prosthesis. Functional results were significantly better for the rotating hinge knee prostheses. Therapeutic study, level IV (case series).


Mitogen-activated protein kinase inhibitors reduce the nuclear accumulation of phosphorylated Smads by inhibiting Imp 7 or Imp 8 in HepG2 cells.
The transforming growth factor (TGF)-β/Smad signaling pathway is involved in hepatocellular carcinoma development. Smad2 and Smad3 are phosphorylated following TGF-β1 stimulation and subsequently oligomerize with Smad4 to form the Smad2/3/4 complex, which translocates into the nucleus and regulates target genes, including plasminogen activator inhibitor type 1 (PAI1). Importin (Imp)7 and Imp8 are responsible for transporting phosphorylated (p)Smad2/3 and Smad4 into the nucleus. In our previous study, it was demonstrated that mitogen-activated protein kinase (MAPK) inhibitors, including inhibitors of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 could inhibit the transcription of PAI1, but ERK inhibitor had no significant effect on the phosphorylation of Smad2/3, and the formation of Smad2/3/4 complexes, which was different from the effect of JNK or p38 inhibitor. We hypothesized that MAPK inhibitors, particularly ERK inhibitor, reduced the transport of Smads into the nucleus by affecting Imp7 and Imp8. To confirm this hypothesis, HepG2 cells were incubated with different MAPK inhibitors for 5 h and subsequently stimulated with TGF-β1 for 1 h. Next, the intracellular locations of Smads (pSmad2C, pSmad2L, pSmad3C, pSmad3L and Smad4) and Imp7/8 were detected using immunofluorescence staining assays, and the expression of Imp7/8 was investigated using immunoblotting. It was revealed that JNK or p38 inhibitor decreased the phosphorylation of Smad2C, Smad2L and Smad3L, and affected their nuclear accumulation. Although only inhibiting the phosphorylation of Smad2C, ERK inhibitor affected the nuclear accumulation of pSmad2C, pSmad2L, pSmad3C and pSmad3L. The three MAPK inhibitors attenuated the nuclear distribution of Smad4, and the expression and nuclear accumulation of Imp7. ERK and JNK inhibitors attenuated the expression and nuclear accumulation of Imp8. Thus, the results of the present study suggest that MAPK inhibitors, particularly ERK inhibitor, modulate the nuclear accumulation of Smads via the inhibition of Imp 7/8.


Stem cell application for osteoarthritis in the knee joint: A minireview.
Knee osteoarthritis is a chronic, indolent disease that will affect an ever increasing number of patients, especially the elderly and the obese. It is characterized by degeneration of the cartilage substance inside the knee which leads to pain, stiffness and tenderness. By some estimations in 2030, only in the United States, this medical condition will burden 67 million people. While conventional treatments like physiotherapy or drugs offer temporary relief of clinical symptoms, restoration of normal cartilage function has been difficult to achieve. Moreover, in severe cases of knee osteoarthritis total knee replacement may be required. Total knee replacements come together with high effort and costs and are not always successful. The aim of this review is to outline the latest advances in stem cell therapy for knee osteoarthritis as well as highlight some of the advantages of stem cell therapy over traditional approaches aimed at restoration of cartilage function in the knee. In addition to the latest advances in the field, challenges associated with stem cell therapy regarding knee cartilage regeneration and chondrogenesis in vitro and in vivo are also outlined and analyzed. Furthermore, based on their critical assessment of the present academic literature the authors of this review share their vision about the future of stem cell applications in the treatment of knee osteoarthritis.


Making choices at menopause.
Menopause is a time of change in women's lives and an opportunity for health promotion. Women can present with a range of symptoms and concerns. The aim of this article is to review the key components of the assessment of women who are perimenopausal or menopausal, and to discuss the benefits and risks of hormonal and non-hormonal treatments, thereby assisting clinicians to appropriately manage these women. The majority of women who are symptomatic can be offered menopausal hormone therapy (MHT). To ensure women can make an informed choice about whether to use MHT, it is important for them to receive balanced and accurate information about the benefits and risks. For women with contraindications to MHT, or women who prefer not to use hormones, non-hormonal and complementary options may be considered. New non-hormonal options are likely to be available in the future.