Novel inhibitors against the transglutaminase-catalysed crosslinking of lens proteins

We had previously reported that the protective effect of taurine against indomethacin-induced gastric mucosal injury was due to its antioxidant effects, which inhibited lipid peroxidation and neutrophil activation. In this study, we examined the effect of taurine on reducing the inflammatory parameters of trinitrobenzene sulfonic acid (TNBS)-induced inflammatory bowel disease (IBD) in rats. In order to induce IBD, ethanolic TNBS was given to rats intracolonically. Then they received 500 mg/kg/day of taurine orally and were sacrificed one week after IBD induction. While ulceration and inflammation of distal colon with formation of granuloma in the vehicle-treated IBD rats two days after administration of TNBS were observed, treatment with taurine ameliorated colonic damage and decreased the incidence of diarrhea and adhesion. Also, colon weight as an index of tissue edema, which was markedly increased in the IBD rats, became significantly lower after taurine treatment. Myeloperoxidase (MPO) activity in the vehicle-treated IBD rats was substantially increased, compared with that of normal control. The taurine-treated animals significantly reduced MPO activity (35% lower) when compared with that of the vehicle-treated animals. Taurine treatment decreased both basal and formyl-methionyl leucyl phenylalanine-stimulated reactive oxygen generation from colonic tissue in the IBD rats. These results suggest that the administration of taurine reduce the inflammatory parameters in this IBD rat model by increasing defending capacity against oxidative damage.     

A practical approach to crosslinking

The various aspects of chemical crosslinking are addressed. Crosslinker reactivity, specificity, spacer arm length and solubility characteristics are detailed. Considerations for choosing one of these crosslinkers for a particular application are given as well as reaction conditions and practical tips for use of each category of crosslinkers.     

A systematic approach to self-medication in older people

This aim of this study was to evaluate the effectiveness of an individualized education programme on older patients' knowledge of prescribed medication. A questionnaire for assessment of medication knowledge was used to collect data from 15 patients before and after an individualized education programme which lasted approximately 2 weeks. Following the collection of pre-intervention data, an individualized education programme was developed for each patient. In addition to verbal instruction, each patient was provided with written information on his/her drug regimen. As part of the education programme each patient also had an opportunity to self-administer his/her medication. The results of the study showed that all participants scored higher in the post-test than the pre-test, although the degree of improvement varied between participants. The average increase in knowledge was 26%. Patient satisfaction with the education programme was also assessed using a questionnaire. Results suggested that there was a very high level of satisfaction with the programme.     

Microdomains of GPI-anchored proteins in living cells revealed by crosslinking

There is some discussion as to whether glycosyl-phosphatidylinositol(GPI)-anchored proteins occur in microdomains in the cell membrane. These putative microdomains have been implicated in processes such as sorting in polarized cells and signal transduction. Complexes enriched in GPI-anchored proteins, cholesterol and glycosphingolipids have been isolated from cell membranes by using non-ionic detergents: these complexes were thought to represent a clustered arrangement of GPI-anchored proteins. However, results obtained when clustering of GPI-anchored proteins induced by antibodies or by detergents was prevented support the idea of a dispersed surface distribution of GPI-anchored proteins at steady state. Here we use chemical crosslinking to show that membrane microdomains of a GPI-anchored protein exist at the surface in living cells. This clustering is specific for the GPI-anchored form, as two transmembrane forms bearing the same ectodomain do not form oligomers. Depletion of membrane cholesterol causes the clustering of GPI-anchored proteins to break up, whereas treatment of cells with detergent substantially increases the size of the complexes. We find that in living cells these GPI-anchored proteins reside in microdomains consisting of at least 15 molecules, which are much smaller than those seen after detergent extraction.     

A social-cognitive skills approach to the successful modification of hypnotic susceptibility.

120 undergraduates who were low and medium in hypnotic susceptibility were administered cognitive strategy and instructional set information and practiced responding to test suggestions to enhance susceptibility. Those in 1 modification treatment received this information both from the experimenter and by observing a videotaped female who responded successfully to suggestions and reported on the cognitive strategies she used to do so. Those in a 2nd modification treatment received the information and practice but were not exposed to the model. Low and medium susceptibles in a 3rd condition (practice alone) received a hypnotic induction procedure and practice suggestions but neither modification information nor modeling. No-treatment controls performed a filler task. All Ss were posttested on 2 susceptibility scales. Results show that information plus modeling produced significantly greater increments on all objective and subjective indices of susceptibility on both posttests than did practice-alone or control treatments. Susceptibility increments in the information without model treatment always fell between those of the model and practice-alone treatments. In the modeling treatment, over half of the initial low susceptibles and over two-thirds of the initial medium susceptibles scored as high susceptibles on both posttests. Findings provide strong support for a social-cognitive skill formulation of hypnotic susceptibility. (49 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

A combinatorial approach to chemical modification of subtilisin Bacillus lentus

The reaction between methanethiosulfonate reagents and cysteine mutants of subtilisin is quantitative and can be used to prepare chemically modified mutant enzymes (CMMs) with novel properties. The virtually unrestricted structural variations possible for CMMs presents a preparative and screening challenge. To address this, a rapid combinatorial method for preparing and screening the activities of CMMs has been developed.     

A probabilistic approach to evaluate creep and fatigue damage in critical components

The subject of reliability assessment of plant components is of considerable importance to maintenance and operations engineers. There exist different degradation mechanisms such as creep and fatigue in the plant components operating under conditions of high pressure and temperature. Hence, the safety issue of these components must be addressed by a realistic reliability model so that the frequency of in-service inspection activities can be optimized. In this work, a probabilistic module based on first order reliability method and Markov model is used in order to calculate different failure state probabilities of an in-service thermal power plant pipe-bend. This procedure helps in online risk assessment of critical plant components. The method of determination partial safety factors of the above component for optimum design based on a target reliability level is also discussed.     

A systematic approach to definitive planning for osseointegrated implant prostheses

Thoughtful design selection is crucial for the perpetual success of any dental implant restoration. This article reviews treatment considerations specific to the postsurgical presentation of the implant patient. Deviations from the originally planned design may be necessary at this time. Indications for the selection of specific prosthetic modalities are offered in a sequence of paradigms to support the cognitive skills of the inexperienced clinician.     

A systematic molecular genetic approach to study mammalian germline development

It is difficult to study gene expression in mammalian embryonic germ cells as PGCs constitute only a minor proportion of the mouse embryo. We have overcome this problem by using a novel combination of established molecular and transgenic approaches. A line of mice has been generated in which the cells of the germ lineage express the beta-galactosidase reporter gene during embryogenesis. Using this line, germ cells have been purified to near homogeneity from embryos at discrete stages during germline development by use of a stain for beta-gal activity and a fluorescence activated cell sorter. Subsequently, cDNA libraries have been constructed from each germ cell population using a modified lone-linker PCR strategy. These combined cDNA libraries represent genes expressed in PGCs during mammalian germline development. To facilitate a molecular genetic approach to studying mammalian germline development, these cDNA libraries will be pooled to form an arrayed, addressed reference embryonic germ cell cDNA library. In parallel with large-scale cDNA sequencing efforts; genes that are differentially expressed in germ cells will be identified by screening the reference library with probes generated by subtractive hybridization. Complementary DNAs identified using this approach will be analyzed by sequencing, database comparison, genomic mapping and in situ hybridization to ascertain the potential functional importance of each gene to germline development. In addition to providing a wealth of novel information regarding patterns of gene expression during mammalian germline development, these results will form the basis for future experiments to determine the function of these genes in this process.     

Covalent modification of PML by the sentrin family of ubiquitin-like proteins

PML, a RING finger protein with tumor suppressor activity, has been implicated in the pathogenesis of acute promyelocytic leukemia that arises following a reciprocal chromosomal translocation that fuses the PML gene with the retinoic acid receptor alpha (RARalpha) gene. Immunocytochemical analysis has demonstrated that PML is co-localized with a novel ubiquitin-like protein in the nuclear bodies, which could be disrupted by the PML-RARalpha fusion protein. The physical nature of this co-localization is unknown. Using a COS cell expression system, we show that PML is covalently modified by all three members of the sentrin family of ubiquitin-like proteins. Covalent modification of PML requires the conserved Gly residue near the C termini of sentrin proteins. Sentrinization of PML is highly specific because neither NEDD8 nor ubiquitin could modify PML. Similar specificity is also observed for the covalent modification of RanGAP1 by the sentrin member of ubiquitin-like proteins. These observations highlight the fine substrate specificity of the sentrinization pathway. In acute promyelocytic leukemia, two forms of PML-RARalpha fusion proteins have been reported. Remarkably, both forms of PML-RARalpha fusion proteins could not be sentrinized. Thus differential sentrinization of PML and PML-RARalpha could play an important role in regulating the biological function of PML and in the pathogenesis of acute promyelocytic leukemia.     

Molecular barriers to biomaterial thrombosis by modification of surface proteins with polyethylene glycol

For cardiovascular biomaterials, thrombosis, thromboembolism and vascular graft occlusion are believed to be precipitated by the adsorption of proteins containing adhesive ligands for platelets. Polyethylene-glycol-diisocyanate (PEG-diisocyanate, 3400 MW) may potentially react with protein amines to form molecular barriers on adsorbed proteins on biomaterials, thereby masking adhesive ligands and preventing acute surface thrombosis. To test this notion, PE, PTFE, and glass microconduits were pre-adsorbed with fibrinogen and treated with PEG-diisocyanate, non-reactive PEG-dihydroxyl, or remained untreated. Following perfusion of 111In-labeled platelets in whole human blood for 1 min (wall shear rate = 312 s(-1)), PEG-diisocyanate treated surfaces experienced 96% (PE), 97% (PTFE) and 94% (glass) less platelet deposition than untreated surfaces. Similar reductions were seen for PEG-diisocyanate versus PEG-dihydroxyl treatment. Low shear perfusions of plasma for 1 h prior to blood contact did not reduce the inhibitory effect of PEG-diisocyanate. Platelet adhesion onto collagen-coated glass coverslips and platelet deposition onto preclotted Dacron were also reduced by treatment with PEG-diisocyanate (93 and 91%, respectively). Protein-reactive PEG may thus have utility in forming molecular barriers on surface-associated proteins to inhibit acute thrombosis on cardiovascular biomaterials.     

A systematic approach to the development of a rational malaria treatment policy in Zambia

The blood-brain barrier (BBB) is a functional characteristic of the cerebral microvasculature that determines which molecules can travel from the bloodstream to the brain parenchyma and vice versa. During the last few decades, we have investigated the contribution of vascular extracellular matrix (ECM) to BBB function. The present review analyzes our findings in the light of recent data on age-related brain function alterations and discusses the potential contribution of BBB function impairment to the processes that determine these alterations.     

工频感应电炉改造方案的探讨

分析了540 kW工频感应电炉3#感应器寿命短的原因,提出改进感应器接线方式和炉体前室结构,达到延长设备使用寿命和节能降耗的目的。     

工频感应电炉改造方案的探讨

分析了540kw工频感应电炉3^#感应器寿命短的原因，提出改进感应器接线方式和炉体前室结构，达到延长设备使用寿命和节能降耗的目的。     

Use of subjective and nonsubjective methodologies to evaluate lens radiation damage in exposed populations--an overview

The general epidemiological acceptability of prevalence, or incidence, for assessing risk of radiation cataract development has dictated an almost exclusive dependence on cataract onset as a measure of cataractogenicity for given doses of radiation. The advent of instrumentation capable of acquiring images amenable to quantitative analyses offers the possibility of exploiting "relative opacification" as an added, if not exclusive, parameter. This development is particularly important in efforts to assess populations such as that in the Altai, which are temporally far removed from their exposure and among whom there exists a large subset with extant cataracts. The new technologies, Scheimpflug and retroillumination imaging, combined with the application of the appropriate analytical algorithms can not only provide quantitative and nonsubjective assessment of lens transparency, but also serve as a means to immortalize the state of the pathology at the time of acquisition. Highly relevant to the assessment of an aging exposed population is the use of lens epithelial fragments as potential dosimeters. The material is routinely available as a result of cataract extraction procedures and is amenable to the application of a modified micronucleus (MN) assay. The MN assay in the lens has tremendous advantages over its use in other tissues for a number of reasons, not least of which is that lens MNs are extremely long-lived. Given the relative ease of application and its potential as a radiation bioindicator, the lens MN assay should be considered in any follow-up of populations exposed to ionizing radiation.     

A systematic approach for interpreting MR images of the seizure patient

A systematic approach needs to be used to review MR scans in epilepsy patients to avoid the common pitfalls engendered by the subtle nature of many epileptogenic lesions. One should always evaluate the hippocampus regardless of other MR findings to avoid missing dual abnormalities. False-positive and false-negative diagnosis of hippocampal sclerosis can be avoided by evaluating the hippocampus after correcting for head rotation [by assessing the internal auditory canals and atria). Periventricular heterotopia can be successfully diagnosed by systematically studying the periventricular regions, especially those adjacent to the atria of the lateral ventricles. Gray matter lateral to the ventricles (excluding the caudate nucleus) is always an abnormal finding. Sulcal and cortical morphologic abnormalities are particularly difficult to diagnose unless a high index of suspicion is maintained. Cortical thickening is indicative of a developmental anomaly and should be screened in an organized manner. Because epilepsy is generally a cortical process, one must search for subtle cortical abnormalities, including focal atrophic abnormalities and lesions without mass effect. Diligence will offer its own rewards.