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PubMed1996

A major allergen of lymphatic filarial nematodes is a parasite homolog of the gamma-glutamyl transpeptidase.

BACKGROUND: Bm2325, a major IgE-inducing antigen of the filarial parasite Brugia malayi has been implicated in the pathology of tropical pulmonary eosinophilia (TPE), a pulmonary syndrome thought to result from hypersensitivity to microfilariae. MATERIALS AND METHODS: Affinity-purified IgE to Bm2325 from patients with TPE was used to identify a complementary DNA (cDNA) from a B. malayi expression library. Sequence analysis of the cDNA revealed a hitherto unknown parasite protein. Immunoblotting of the recombinant filarial protein using sera of patients with TPE determined its IgE-binding capacity. Reactivity to human lung epithelial cell proteins was analyzed using murine anti-Bm2325 antibodies and serum from patients with TPE. RESULTS: The predicted protein is a homolog of the entire precursor of the gamma-glutamyl transpeptidase (gamma-GT), a key enzyme in the synthesis and degradation of glutathione. The filarial precursor encodes both the heavy (H) and the light (L) chain subunits and shares structural similarities with the mammalian enzymes. The Bm2325 allergen was identified as the homolog of the enzyme light chain subunit. Murine antibodies against the recombinant parasite gamma-GT cross-reacted with the human enzyme present in human airway epithelial cells, and human gamma-GT is a target of antibodies present in the serum of patients with TPE. CONCLUSION: Molecular mimicry between the parasite gamma-GT homolog and the host membrane-bound gamma-GT present in lung epithelial cells likely contributes to the pathogenesis observed in tropical pulmonary eosinophilia.

Amino Acid Sequence↗
PubMed1991

Improved interpretation of complex chromosomal rearrangements by combined GTG banding and in situ suppression hybridization using chromosome-specific libraries and cosmid probes.

Chromosome aberrations of a hypodiploid ovarian carcinoma cell line (modal chromosome number 38) having a complex karyotype were analyzed using biotinylated DNA library probes that specifically hybridize to chromosomes 3, 6, 7, 8, 11, 13, and 16 from telomere (pter) to telomere (qter). A series of cosmid probes localized to the short arm of chromosome 16 were used to further investigate one of the two aberrant chromosomes 16 present in this cell line. The competitive in situ suppression (CISS) hybridization of DNA-libraries was mostly performed subsequent to GTG-banding of the same metaphase cell in order to interpret the hybridization signals optimally. This combined approach made it possible to detect the origin of chromosomal material that could not be identified using GTG-banding. Furthermore, the in situ hybridization techniques appeared to be helpful in the characterization of complex translocations and for accurate breakpoint determination.

Adenocarcinoma↗
PubMed1998

Bacillus cereus in a whey process.

A cheese dairy and its whey manufacturing line were examined for Bacillus cereus. Colonies typical of B. cereus were detected in 120 (17%) samples out of 720 analysed. Only 3% of the sampled raw milk contained B. cereus ( > or = 10 cfu ml(-1)) whereas in evaporated whey concentrate B. cereus was present in 76% of the samples. Nitrate reductase negative and weakly casein hydrolysis isolates were rare in raw milk and the early parts of the process but these defective biotypes became increasingly frequent towards the end of the whey process. The composition of whole cell fatty acids of B. cereus isolates originating from the whey part of the process was different from that of the type strain and of the isolates originating from the raw materials of cheese making. The B. cereus strains in concentrated whey were 100% similar to the type strain in 16S rDNA sequence (500 bp) although they were not or only poorly recognized as B. cereus by a commercial whole cell fatty acid library. All of B. cereus isolates in raw milk were sensitive to one or more of the B. cereus group phages (n = 17) whereas 43% of the isolates from the whey process were sensitive to none. None of the 23 strains originating from the whey processing lines grew at < or = 8 degrees C. although strains with minimum growth temperatures of 5.3 degrees C and 7.0 degrees C were present in the raw materials. Our results indicate that the B. cereus population of the warm ( > 30 degrees C) parts of the cheese dairy process was separate from that of cold (2 degrees C to 4 degrees C) part of the process.

Bacillus cereus↗
PubMed2003

Novel heparan mimetics potently inhibit the scrapie prion protein and its endocytosis.

During prion diseases the normal prion protein PrP(C) is refolded into an abnormal conformer PrP(Sc). We have studied the PrP(Sc) inhibiting activity of a library of synthetic heparan mimetic (HM) biopolymers. HMs are chemically derived dextrans obtained by successive substitutions with carboxymethyl, benzylamide, and sulfate groups on glucose residues. Some HMs eliminated PrP(Sc) from prion-infected cells after a 5 day course at 100 ng/ml and were 15 x potent than pentosan sulfate in this system. The anti-PrP(Sc) activity of HMs correlated with the degree of sulfation but was increased by benzylamidation. HMs did not reduce the synthesis of PrP(C) nor its attachment to lipid rafts, but instead blocked its conversion into PrP(Sc). The anti-PrP(Sc) HMs also prevented the uptake of prion rods by cultured cells. HMs may thus block the interaction of PrP(Sc) with a putative cellular receptor, possibly heparan sulfate. HMs provide an attractive chemical approach for the synthesis of TSE therapeutic and prophylactic reagents.

Animals↗
PubMed2002

Application of complex macromolecular architectures for advanced microelectronic materials.

The distinctive features of well-defined, three-dimensional macromolecules with topologies designed to enhance solubility and amplify end-group functionality facilitated nanophase morphologies in mixtures with organosilicates and ultimately nanoporous organosilicate networks. Novel macromolecular architectures including dendritic and star-shaped polymers and organic nanoparticles were prepared by a modular approach from several libraries of building blocks including various generations of dendritic initiators and dendrons, selectively placed to amplify functionality and/or arm number, coupled with living polymerization techniques. Mixtures of an organosilicate and the macromolecular template were deposited, cured, and the phase separation of the organic component, organized the vitrifying organosilicate into nanostructures. Removal of the sacrificial macromolecular template, also denoted as porogen, by thermolysis, yielded the desired nanoporous organosilicate, and the size scale of phase separation was strongly dependent on the chain topology. These materials were designed for use as interlayer, ultra-low dielectric insulators for on-chip applications with dielectric constant values as low as 1.5. The porogen design, chemistry and role of polymer architecture on hybrid and pore morphology will be emphasized.

Electronics↗
PubMed1997

Long PCR facilitates concise cloning and sequencing with a minimal tiling set of templates.

We demonstrate a rapid cloning and sequencing strategy for kilobase-size DNA segments using DNase I and long PCR. In a single-tube protocol, deletions were formed in a plasmid insert by two enzymatic cuts, one at a fixed site and one at random. The doubly cut molecules were recircularized to generate a library of plasmids carrying deletions of various sizes and transformed into E. coli. The plasmid inserts were directly amplified from transformant colonies by long PCR and sized on a high-resolution agarose gel. A minimal tiling set, selected from the amplified material, was used directly as templates for long-read sequencing. The system is useful for inserts up to about 3.5 kb for de novo sequencing (both strands) or 6 kb for confirmatory sequencing (one strand).

Cloning, Molecular↗
PubMed2005

Diaminocarbene- and Fischer-carbene complexes of palladium and nickel by oxidative insertion: preparation, structure, and catalytic activity.

Oxidative insertion of [Pd(PPh3)4] or [Ni(cod)2]/PPh3 into the C-Cl bond of various 2-chloroimidazolinium- and other -amidinium salts affords metal-diaminocarbene complexes in good to excellent yields. This procedure is complementary to existing methodology in which the central metal does not change its oxidation state, and therefore allows to incorporate carbene fragments that are difficult to access otherwise. The preparation of a variety of achiral as well as enantiomerically pure, chiral metal-NHC complexes (NHC = N-heterocyclic carbene) and metal complexes with acyclic diaminocarbene ligands illustrates this aspect. Furthermore it is shown that oxidative insertion also paves a way to prototype Fischer carbenes of Pd(II). Since the required starting materials are readily available from urea- or thiourea derivatives, this novel approach allows for substantial structural variations of the ligand backbone. The catalytic performance of the resulting library of nickel- and palladium-carbene complexes has been evaluated by applications to prototype Suzuki-, Heck-, and Kumada-Corriu cross-coupling reactions as well as Buchwald-Hartwig aminations. It was found that even Fischer carbenes show appreciable catalytic activity. Moreover, representative examples of all types of neutral and cationic metal-carbene complexes formed in this study have been characterized by X-ray crystallography.

Journal Article↗
PubMed1998

Affinity capillary electrophoresis in biomolecular recognition.

Affinity capillary electrophoresis is a new method for studies of biomolecular recognition. Applications reported in the literature include chiral separation of racemic biomolecules, measurement of binding constants, estimation of kinetic on- and off-rate constants, determination of binding stoichiometries (a useful tool in examining electrostatic interactions), estimation of effective charges and molecular weights of proteins, characterization of enzymatic activities and library screening for tight-binding drug candidates in solution. This technique demands only small amounts of sample (nanolitre injection volumes, picograms of proteins), involves no radiolabelled materials or chemically immobilized ligands, and does not require changes in spectroscopic characteristics upon binding. This paper reviews the most recent applications of affinity capillary electrophoresis and its use in the analysis of biomolecules.

Electrophoresis, Capillary↗
PubMed2005

Novel actinobacteria from marine sponges.

Actinobacteria exclusively within the sub-class Acidimicrobidae were shown by 16S rDNA community analysis to be major components of the bacterial community associated with two sponge species in the genus Xestospongia. Four groups of Actinobacteria were identified in Xestospongia spp., with three of these four groups being found in both Xestospongia muta from Key Largo, Florida and Xestospongia testudinaria from Manado, Indonesia. This suggests that these groups are true symbionts in these sponges and may play a common role in both the Pacific and Atlantic sponge species. The fourth group was found only in X. testudinaria and was a novel assemblage distantly related to any previously sequenced actinobacterial clones. The only actinobacteria that were obtained in initial culturing attempts were Gordonia, Micrococcus and Brachybacterium spp., none of which were represented in the clone libraries. The closest cultured actinobacteria to all the Acidimicrobidae clones from Xestospongia spp. are 'Microthrix parvicella' and Acidimicrobium spp. Xestospongia spp. can now be targeted as source material from which to culture novel Acidimicrobidae to investigate their potential as producers of bioactive compounds. Isolation of sponge-associated Acidimicrobidae will also make it possible to elucidate their role as sponge symbionts.

Actinobacteria↗
PubMed2000

Stereo-random synthesis of highly functionalized proline analogues by azomethine cycloaddition.

Highly substituted proline analogues were synthesized on Wang-resin bearing bisprotected histidine as starting material. The proline analogues (1,5-diazabicyclo[3.3.0]octane-2-carboxylic acid) were generated by 1,3-dipolar cycloaddition of azomethine ylides with maleimides, thus creating a library of maximum stereochemical diversity. Every compound set with the same empirical formula can theoretically consist of four diastereomers and can be tested in biological assays as mixture. Additionally different methods for the acylation of the proline nitrogen were evaluated.

Bridged Bicyclo Compounds, Heterocyclic↗
PubMedDate not supplied

Selective nucleophilic chemistry in the synthesis of 5-carbamoyl-3-sulfanylmethylisoxazole-4-carboxylic acids.

The solution-phase syntheses of 5-carbamoyl-3-sulfanylmethylisoxazole-4-carboxylic acids were accomplished from dimethyl 3-chloromethylisoxazole-4,5-dicarboxylate by selective nucleophilic chemistry. For example, treatment of this trifunctionalized core with 3-bromobenzylamine and subsequent X-ray analysis identified the sole product as methyl 5-(3-bromobenzylcarbamoyl)-3-chloromethylisoxazole-4-carboxylate. Subjecting this amide/ester to thiophenol in the presence of 1 N NaOH completed the two-step transformation of this versatile starting material to the targeted 5-carbamoyl-3-sulfanylmethylisoxazole-4-carboxylic acid. Employing various amines and thiophenols, this chemistry was applied in the generation of a 90-compound library of druglike isoxazoles.

Carboxylic Acids↗
PubMedDate not supplied

An integrated instrument control and informatics system for combinatorial materials research.

The use of high-throughput synthesis and characterization techniques is increasingly prevalent in materials science research. We describe the London University Search Instrument, a research apparatus designed for the high-throughput synthesis and characterization of thick-film sample libraries of ceramic compounds. The instrument is constructed largely from commodity components, which pose particular engineering challenges for achieving the automated operation required for efficient high-throughput experimentation. This paper describes the architecture and implementation of the software system that provides integrated instrument control and data management functions.

Journal Article↗
PubMed1999

Substance abuse studies and prevention efforts among Arabs in the 1990s in Israel, Jordan and the Palestinian Authority--a literature review.

This paper is the result of a collaborative project of Israeli, Jordanian and Palestinian scientists gathered to reveal the current extent of substance abuse and efforts at prevention among Arabs in Israel, Jordan and the Palestinian Authority territories, in order to identify needs and suggest future collaborative activities and directions for regional cooperation. The article provides data and covers the current state of substance abuse prevention and research among Moslems, Christians and Druze in the trilateral region in the 1990s by reviewing prevention materials and studies published in the professional literature, as well as in reports and Doctoral and Master's theses in Arabic, which have been located in academic libraries and other institutions, in the framework of a comprehensive search. This manuscript is the first to summarize Jordanian and Palestinian findings in the substance abuse domain. The review shows that most of the Israeli research in the Arab sector deals with alcohol use among youth, that the majority of Jordanian studies focus on illicit drug use, that the research among Palestinians is in its infancy, and that comprehensive prevention programs are lacking in the trilateral region. It describes the key results of most of the 12 Israeli studies among Arabs, 11 Jordanian studies and four Palestinian studies. It reveals that drug abuse among Israeli Arab students is probably more prevalent than among Jewish adolescents, that the typical Jordanian drug addict has a higher level of education than the typical Palestinian drug addict, and that the Palestinian is more likely to be a multiple drug user. Recommendations for future activities include organization of a regional collaborative workshop in order to establish data collection systems for basic statistics relevant to drug abuse and development of comprehensive prevention programs, as well as studies in the substance abuse domain concerning knowledge, attitudes and behavior among the general Arab population.

Arabs↗
PubMed2006

Publishing trends in Chinese medicine and related subjects documented in WorldCat.

BACKGROUND: Chinese medicine (CM) has been the subject of increasing interest in the past 30 years, both as a discipline and in the larger context of alternative medicine. It has steadily been accepted by and integrated into the medical and health-care fields in many countries. OBJECTIVE: This study aims to gain an overview of how CM has been interpreted and presented to the world outside China and to identify emerging trends. METHODS: This study is designed to analyse the publishing trends of CM and related subjects in all languages except Chinese, ranging from books and serials to audio-visual and electronic resources found in WorldCat, the world's largest bibliographic database produced by OnLine Computer Library Center (OCLC). RESULTS: The findings showed a flourishing growth of publications in CM and related subjects beginning in the 1970s with greater coverage on acupuncture. The materials in English language constitute the major portion of total output. CONCLUSION: We conclude that Chinese medicine has steadily gained recognition in the world based on the analysis of publication records. The translation of original works and analysis of journal literature and conference proceedings on Chinese medicine merit further study.

Acupuncture↗
PubMed2000

Laser capture microdissection in pathology.

The molecular examination of pathologically altered cells and tissues at the DNA, RNA, and protein level has revolutionised research and diagnostics in pathology. However, the inherent heterogeneity of primary tissues with an admixture of various reactive cell populations can affect the outcome and interpretation of molecular studies. Recently, microdissection of tissue sections and cytological preparations has been used increasingly for the isolation of homogeneous, morphologically identified cell populations, thus overcoming the obstacle of tissue complexity. In conjunction with sensitive analytical techniques, such as the polymerase chain reaction, microdissection allows precise in vivo examination of cell populations, such as carcinoma in situ or the malignant cells of Hodgkin's disease, which are otherwise inaccessible for conventional molecular studies. However, most microdissection techniques are very time consuming and require a high degree of manual dexterity, which limits their practical use. Laser capture microdissection (LCM), a novel technique developed at the National Cancer Institute, is an important advance in terms of speed, ease of use, and versatility of microdissection. LCM is based on the adherence of visually selected cells to a thermoplastic membrane, which overlies the dehydrated tissue section and is focally melted by triggering of a low energy infrared laser pulse. The melted membrane forms a composite with the selected tissue area, which can be removed by simple lifting of the membrane. LCM can be applied to a wide range of cell and tissue preparations including paraffin wax embedded material. The use of immunohistochemical stains allows the selection of cells according to phenotypic and functional characteristics. Depending on the starting material, DNA, good quality mRNA, and proteins can be extracted successfully from captured tissue fragments, down to the single cell level. In combination with techniques like expression library construction, cDNA array hybridisation and differential display, LCM will allow the establishment of "genetic fingerprints" of specific pathological lesions, especially malignant neoplasms. In addition to the identification of new diagnostic and prognostic markers, this approach could help in establishing individualised treatments tailored to the molecular profile of a tumour. This review provides an overview of the technique of LCM, summarises current applications and new methodical approaches, and tries to give a perspective on future developments. In addition, LCM is compared with other recently developed laser microdissection techniques.

Cell Adhesion↗
PubMed2005

Ethnic disparities: control of glycemia, blood pressure, and LDL cholesterol among US adults with type 2 diabetes.

OBJECTIVE: To examine ethnic disparities in the quality of diabetes care among adults with diabetes in the US through a systematic qualitative review. DATA SOURCES: Material published in the English language was searched from 1993 through June 2003 using PubMed, Web of Science, Cumulative Index to Nursing and Allied Health, the Cochrane Library, Combined Health Information Database, and Education Resources Information Center. STUDY SELECTION AND DATA EXTRACTION: Studies of patients with diabetes in which at least 50% of study participants were ethnic minorities and studies that made ethnic group comparisons were eligible. Research on individuals having prediabetes, those <18 years of age, or women with gestational diabetes were excluded. Reviewers used a reproducible search strategy. A standardized abstraction and grading of articles for publication source and content were used. Data on glycemia, blood pressure, and low-density lipoprotein cholesterol (LDL-C) were extracted in patients with diabetes. A total of 390 studies were reviewed, with 78 meeting inclusion criteria. DATA SYNTHESIS: Ethnic minorities had poorer outcomes of care than non-Hispanic whites. These disparities were most pronounced for glycemic control and least evident for LDL-C control. Most studies showed blood pressure to be poorly controlled among ethnic minorities. CONCLUSIONS: Control of risk factors for diabetes (glycemia, blood pressure, LDL-C) is challenging and requires routine assessment. These findings indicate that additional efforts are needed to promote diabetes quality of care among minority populations.

Adult↗
PubMed2002

Frontal affinity chromatography for the screening of mixtures.

A protein stationary phase for frontal affinity chromatography was prepared, containing biotinylated beta-galactosidase immobilized to controlled pore glass beads via covalently bonded streptavidin. Single microaffinity columns of approximately 30 pmol of active beta-galactosidase were prepared from this material and characterized with a known ligand by frontal analysis. These columns were used to measure the specific interactions between the bound beta-galactosidase and a library of modified beta-galactopyranosides using electrospray mass spectrometry as the means of detection. The library contained 89 entries, each representing 4 diastereomers for a total of 356 library members. A single entry was analysed revealing differential activity among the 4 isomers. The library was grouped into 10 mixtures of 24-40 members each with each mixture infused under frontal chromatographic conditions. This deconvolution procedure led to the identification of 34 entries containing isomers with K(d) values better than 10 microM. A method based on a displacement principle was implemented as a rapid prescreen which served as the basis for a parallel column high throughput screening assay.

Chromatography, Affinity↗