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Sample records for human proinsulin determining

  1. Development and standardization of radioimmunoassay technique for human proinsulin determining and its use in the study of type II diabetes mellitus associated to obesity

    International Nuclear Information System (INIS)

    Nascimento, Martha do

    1996-01-01

    The availability of immunoassay methodology for proinsulin is important to define its physiological and pathophysiological significance in humans. Serum concentration of proinsulin are elevated in patients with type II Diabetes Mellitus (NIDDM) and recently diagnosed Type I, so a raised circulating concentration of proinsulin may serve as an early indicator of β cells dysfunction. recently, in NIDDM the serum concentrations of proinsulin and its B-chain-C-peptide junctional split form, des (31-32), were found to correlate with diastolic blood pressure, a risk factor for cardiovascular disease. The development of a sensitive and specific radioimmunoassay (RIA) methodology for proinsulin has been difficult due to its low concentration in serum and the presence of proinsulin conversion intermediates in fluids and tissues. Also other potentially cross-reactive peptides, including insulin and C-peptide, can interfere in the assay. This work describe a highly specific human proinsulin RIA developed by using biosynthetic human proinsulin (hPI) as immunogen, standard and tracer. (author)

  2. Production of antisera for radioimmunoassay of human proinsulin: preliminary studies

    International Nuclear Information System (INIS)

    Nascimento, M. do; Borghi, V.C.; Bellini, M.H.; Wajchenberg, B.L.

    1990-01-01

    In attempt to the production of anti proinsulin antisera for the radioimmunoassay, five guinea pigs received 50 μg of biosynthetic human proinsulin (Eli Lilly and Company, US) injected subcutaneously. Booster injections were administered at 3 weeks intervals and blood was taken 14 days after the injection. Just after the tenth week, three guinea pigs developed antisera with titers of 1:15.000, 1:1.500 and 1:300 and one of them presented titer lower than 1:300. The evaluation of the specificity of those three antisera showed cross-reactivity only with insulin, which antibodies were not sufficiently sensitive for the assay of endogenous proinsulin from healthy subjects, being only suitable for the measurement of circulating proinsulin in patients with hyper proinsulinemia. Alternative schedules of immunization in order to obtain more sensitive antisera are discussed. (author)

  3. NMR and photo-CIDNP studies of human proinsulin and prohormone processing intermediates with application to endopeptidase recognition

    International Nuclear Information System (INIS)

    Weiss, M.A.; Frank, B.H.; Heiney, R.; Pekar, A.; Khait, I.; Neuringer, L.J.; Shoelson, S.E.

    1990-01-01

    The proinsulin-insulin system provides a general model for the proteolytic processing of polypeptide hormones. Two proinsulin-specific endopeptidases have been defined, a type I activity that cleaves the B-chain/C-peptide junction (Arg 31 -Arg 32 ) and a type II activity that cleaves the C-peptide/A-chain junction (Lys 64 -Arg 65 ). These endopeptidases are specific for their respective dibasic target sites; not all such dibasic sites are cleaved, however, and studies of mutant proinsulins have demonstrated that additional sequence or structural features are involved in determining substrate specificity. To define structural elements required for endopeptidase recognition, the authors have undertaken comparative 1 H NMR and photochemical dynamic nuclear polarization (photo-CIDNP) studies of human proinsulin, insulin, and split proinsulin analogues as models or prohormone processing intermediates. The overall conformation of proinsulin is observed to be similar to that of insulin, and the connecting peptide is largely unstructured. In the 1 H NMR spectrum of proinsulin significant variation is observed in the line widths of insulin-specific amide resonances, reflecting exchange among conformational substrates; similar exchange is observed in insulin and is not damped by the connecting peptide. The aromatic 1 H NMR resonances of proinsulin are assigned by analogy to the spectrum of insulin, and assignments are verified by chemical modification. These results suggest that a stable local structure is formed at the CA junction, which influences insulin-specific packing interactions. They propose that this structure (designated the CA knuckle) provides a recognition element for type II proinsulin endopeptidase

  4. Automated thermometric enzyme immunoassay of human proinsulin produced by Escherichia coli.

    Science.gov (United States)

    Birnbaum, S; Bülow, L; Hardy, K; Danielsson, B; Mosbach, K

    1986-10-01

    We have determined and monitored the production and release of human proinsulin by genetically engineered Escherichia coli cells. Several M9 media samples were analyzed sequentially after centrifugation with the aid of a rapid automated flow-through thermometric enzyme-linked immunosorbent assay (TELISA) system. The response time was 7 min after sample injection and a single assay was complete after 13 min. Insulin concentrations in the range of 0.1-50 micrograms/ml could be determined. The TELISA method correlated well with conventional radioimmunoassay determinations. Standard curves were reproducible over a period of several days even when the immobilized antibody column was stored at 25 degrees C in the enzyme thermistor unit. Thus, immediate assay start up was possible.

  5. Automated thermometric enzyme immunoassay of human proinsulin produced by Escherichia coli

    International Nuclear Information System (INIS)

    Birnbaum, S.; Buelow, L.; Hardy, K.; Danielsson, B.; Mosbach, K.

    1986-01-01

    The authors have determined and monitored the production and release of human proinsulin by genetically engineered Escherichia coli cells. Several M9 media samples were analyzed sequentially after centrifugation with the aid of a rapid automated flow-through thermometric enzyme-linked immunosorbent assay (TELISA) system. The response time was 7 min after after sample injection and a single assay was complete after 13 min. Insulin concentrations in the range of 0.1-50 μg/ml could be determined. The TELISA method correlated well with conventional radioimmunoassay determinations. Standard curves were reproducible over a period of several days even when the immobilized antibody column was stored at 25 0 C in the enzyme thermistor unit. Thus, immediate assay start up was possible

  6. Development and standardization of radioimmunoassay technique for human proinsulin determining and its use in the study of type II diabetes mellitus associated to obesity; Desenvolvimento e padronizacao da tecnica de radioimunoensaio para a determinacao de pro-insulina humana e sua aplicacao no estudo do diabetes mellitus tipo II associado a obesidade

    Energy Technology Data Exchange (ETDEWEB)

    Nascimento, Martha do

    1996-12-31

    The availability of immunoassay methodology for proinsulin is important to define its physiological and pathophysiological significance in humans. Serum concentration of proinsulin are elevated in patients with type II Diabetes Mellitus (NIDDM) and recently diagnosed Type I, so a raised circulating concentration of proinsulin may serve as an early indicator of {beta} cells dysfunction. recently, in NIDDM the serum concentrations of proinsulin and its B-chain-C-peptide junctional split form, des (31-32), were found to correlate with diastolic blood pressure, a risk factor for cardiovascular disease. The development of a sensitive and specific radioimmunoassay (RIA) methodology for proinsulin has been difficult due to its low concentration in serum and the presence of proinsulin conversion intermediates in fluids and tissues. Also other potentially cross-reactive peptides, including insulin and C-peptide, can interfere in the assay. This work describe a highly specific human proinsulin RIA developed by using biosynthetic human proinsulin (hPI) as immunogen, standard and tracer. (author) 133 refs., 17 figs., 36 tabs.

  7. Development and standardization of radioimmunoassay technique for human proinsulin determining and its use in the study of type II diabetes mellitus associated to obesity; Desenvolvimento e padronizacao da tecnica de radioimunoensaio para a determinacao de pro-insulina humana e sua aplicacao no estudo do diabetes mellitus tipo II associado a obesidade

    Energy Technology Data Exchange (ETDEWEB)

    Nascimento, Martha do

    1997-12-31

    The availability of immunoassay methodology for proinsulin is important to define its physiological and pathophysiological significance in humans. Serum concentration of proinsulin are elevated in patients with type II Diabetes Mellitus (NIDDM) and recently diagnosed Type I, so a raised circulating concentration of proinsulin may serve as an early indicator of {beta} cells dysfunction. recently, in NIDDM the serum concentrations of proinsulin and its B-chain-C-peptide junctional split form, des (31-32), were found to correlate with diastolic blood pressure, a risk factor for cardiovascular disease. The development of a sensitive and specific radioimmunoassay (RIA) methodology for proinsulin has been difficult due to its low concentration in serum and the presence of proinsulin conversion intermediates in fluids and tissues. Also other potentially cross-reactive peptides, including insulin and C-peptide, can interfere in the assay. This work describe a highly specific human proinsulin RIA developed by using biosynthetic human proinsulin (hPI) as immunogen, standard and tracer. (author) 133 refs., 17 figs., 36 tabs.

  8. Recombinant human proinsulin from transgenic corn endosperm: solvent screening and extraction studies

    Directory of Open Access Journals (Sweden)

    C. S. Farinas

    2007-09-01

    Full Text Available Recombinant pharmaceutical proteins are being produced in different systems such as bacteria and mammalian cell cultures. The use of transgenic plants as bioreactors has recently arisen as an alternative system offering many practical and economic advantages. However, finding an optimum strategy for the downstream processing (DSP of recombinant proteins from plants still remains a challenge. In this work, we studied the extraction of recombinant human proinsulin (rhProinsulin produced in the endosperm of transgenic corn seeds. An efficient extraction solvent was selected and the effects of temperature, solvent-to-solid ratio, time, and impeller rotational speed on the extraction were evaluated using an experimental design. After an extraction kinetics study, temperature was further evaluated to maximize rhProinsulin concentration in the extracts and to minimize the native corn components carbohydrates, phenolic compounds, and proteins. A high efficiency condition for extracting rhProinsulin with the selected solvent - 50 mM sodium bicarbonate buffer pH 10.0 and 5 mM DTT - was an extraction time of 2 h at a solvent-to-solid ratio of 10:1 and 25º C. The maximum rhProinsulin concentration in the extracts at that condition was 18.87 mg l-1 or 0.42% of the total soluble protein. These values are within the range in which the production of pharmaceutical proteins in plants can be competitive with other expression systems. The results presented provide information for the development of an additional production platform for the hormone insulin.

  9. Induction of indoleamine 2, 3-dioxygenase in human dendritic cells by a cholera toxin B subunit-proinsulin vaccine.

    Directory of Open Access Journals (Sweden)

    Jacques C Mbongue

    Full Text Available Dendritic cells (DC interact with naïve T cells to regulate the delicate balance between immunity and tolerance required to maintain immunological homeostasis. In this study, immature human dendritic cells (iDC were inoculated with a chimeric fusion protein vaccine containing the pancreatic β-cell auto-antigen proinsulin linked to a mucosal adjuvant the cholera toxin B subunit (CTB-INS. Proteomic analysis of vaccine inoculated DCs revealed strong up-regulation of the tryptophan catabolic enzyme indoleamine 2, 3-dioxygenase (IDO1. Increased biosynthesis of the immunosuppressive enzyme was detected in DCs inoculated with the CTB-INS fusion protein but not in DCs inoculated with proinsulin, CTB, or an unlinked combination of the two proteins. Immunoblot and PCR analyses of vaccine treated DCs detected IDO1mRNA by 3 hours and IDO1 protein synthesis by 6 hours after vaccine inoculation. Determination of IDO1 activity in vaccinated DCs by measurement of tryptophan degradation products (kynurenines showed increased tryptophan cleavage into N-formyl kynurenine. Vaccination did not interfere with monocytes differentiation into DC, suggesting the vaccine can function safely in the human immune system. Treatment of vaccinated DCs with pharmacological NF-κB inhibitors ACHP or DHMEQ significantly inhibited IDO1 biosynthesis, suggesting a role for NF-κB signaling in vaccine up-regulation of dendritic cell IDO1. Heat map analysis of the proteomic data revealed an overall down-regulation of vaccinated DC functions, suggesting vaccine suppression of DC maturation. Together, our experimental data indicate that CTB-INS vaccine induction of IDO1 biosynthesis in human DCs may result in the inhibition of DC maturation generating a durable state of immunological tolerance. Understanding how CTB-INS modulates IDO1 activity in human DCs will facilitate vaccine efficacy and safety, moving this immunosuppressive strategy closer to clinical applications for prevention

  10. Intracellular transport and sorting of mutant human proinsulins that fail to form hexamers.

    Science.gov (United States)

    Quinn, D; Orci, L; Ravazzola, M; Moore, H P

    1991-06-01

    Human proinsulin and insulin oligomerize to form dimers and hexamers. It has been suggested that the ability of prohormones to self associate and form aggregates may be responsible for the sorting process at the trans-Golgi. To examine whether insulin oligomerization is required for proper sorting into regulated storage granules, we have constructed point mutations in human insulin B chain that have been previously shown to prevent formation of insulin hexamers (Brange, J., U. Ribel, J. F. Hansen, G. Dodson, M. T. Hansen, S. Havelund, S. G. Melberg, F. Norris, K. Norris, L. Snel, A. R. Sorensen, and H. O. Voight. 1988. Nature [Lond.]. 333:679-682). One mutant (B10His----Asp) allows formation of dimers but not hexamers and the other (B9Ser----Asp) prevents formation of both dimers and hexamers. The mutants were transfected into the mouse pituitary AtT-20 cells, and their ability to be sorted into regulated secretory granules was compared to wild-type insulin. We found that while B10His----Asp is sorted somewhat less efficiently than wild-type insulin as reported previously (Carroll, R. J., R. E. Hammer, S. J. Chan, H. H. Swift, A. H. Rubenstein, and D. F. Steiner. 1988. Proc. Natl. Acad. Sci. USA. 85:8943-8947; Gross, D. J., P. A. Halban, C. R. Kahn, G. C. Weir, and L. Villa-Kumaroff. 1989. Proc. Natl. Acad. Sci. USA. 86:4107-4111). B9Ser----Asp is targeted to granules as efficiently as wild-type insulin. These results indicate that self association of proinsulin into hexamers is not required for its targeting to the regulated secretory pathway.

  11. Genetic complexity in a Drosophila model of diabetes-associated misfolded human proinsulin.

    Science.gov (United States)

    Park, Soo-Young; Ludwig, Michael Z; Tamarina, Natalia A; He, Bin Z; Carl, Sarah H; Dickerson, Desiree A; Barse, Levi; Arun, Bharath; Williams, Calvin L; Miles, Cecelia M; Philipson, Louis H; Steiner, Donald F; Bell, Graeme I; Kreitman, Martin

    2014-02-01

    Drosophila melanogaster has been widely used as a model of human Mendelian disease, but its value in modeling complex disease has received little attention. Fly models of complex disease would enable high-resolution mapping of disease-modifying loci and the identification of novel targets for therapeutic intervention. Here, we describe a fly model of permanent neonatal diabetes mellitus and explore the complexity of this model. The approach involves the transgenic expression of a misfolded mutant of human preproinsulin, hINS(C96Y), which is a cause of permanent neonatal diabetes. When expressed in fly imaginal discs, hINS(C96Y) causes a reduction of adult structures, including the eye, wing, and notum. Eye imaginal discs exhibit defects in both the structure and the arrangement of ommatidia. In the wing, expression of hINS(C96Y) leads to ectopic expression of veins and mechano-sensory organs, indicating disruption of wild-type signaling processes regulating cell fates. These readily measurable "disease" phenotypes are sensitive to temperature, gene dose, and sex. Mutant (but not wild-type) proinsulin expression in the eye imaginal disc induces IRE1-mediated XBP1 alternative splicing, a signal for endoplasmic reticulum stress response activation, and produces global change in gene expression. Mutant hINS transgene tester strains, when crossed to stocks from the Drosophila Genetic Reference Panel, produce F1 adults with a continuous range of disease phenotypes and large broad-sense heritability. Surprisingly, the severity of mutant hINS-induced disease in the eye is not correlated with that in the notum in these crosses, nor with eye reduction phenotypes caused by the expression of two dominant eye mutants acting in two different eye development pathways, Drop (Dr) or Lobe (L), when crossed into the same genetic backgrounds. The tissue specificity of genetic variability for mutant hINS-induced disease has, therefore, its own distinct signature. The genetic dominance

  12. Effect of genetic variation in a Drosophila model of diabetes-associated misfolded human proinsulin.

    Science.gov (United States)

    He, Bin Z; Ludwig, Michael Z; Dickerson, Desiree A; Barse, Levi; Arun, Bharath; Vilhjálmsson, Bjarni J; Jiang, Pengyao; Park, Soo-Young; Tamarina, Natalia A; Selleck, Scott B; Wittkopp, Patricia J; Bell, Graeme I; Kreitman, Martin

    2014-02-01

    The identification and validation of gene-gene interactions is a major challenge in human studies. Here, we explore an approach for studying epistasis in humans using a Drosophila melanogaster model of neonatal diabetes mellitus. Expression of the mutant preproinsulin (hINS(C96Y)) in the eye imaginal disc mimics the human disease: it activates conserved stress-response pathways and leads to cell death (reduction in eye area). Dominant-acting variants in wild-derived inbred lines from the Drosophila Genetics Reference Panel produce a continuous, highly heritable distribution of eye-degeneration phenotypes in a hINS(C96Y) background. A genome-wide association study (GWAS) in 154 sequenced lines identified a sharp peak on chromosome 3L, which mapped to a 400-bp linkage block within an intron of the gene sulfateless (sfl). RNAi knockdown of sfl enhanced the eye-degeneration phenotype in a mutant-hINS-dependent manner. RNAi against two additional genes in the heparan sulfate (HS) biosynthetic pathway (ttv and botv), in which sfl acts, also modified the eye phenotype in a hINS(C96Y)-dependent manner, strongly suggesting a novel link between HS-modified proteins and cellular responses to misfolded proteins. Finally, we evaluated allele-specific expression difference between the two major sfl-intronic haplotypes in heterozygtes. The results showed significant heterogeneity in marker-associated gene expression, thereby leaving the causal mutation(s) and its mechanism unidentified. In conclusion, the ability to create a model of human genetic disease, map a QTL by GWAS to a specific gene, and validate its contribution to disease with available genetic resources and the potential to experimentally link the variant to a molecular mechanism demonstrate the many advantages Drosophila holds in determining the genetic underpinnings of human disease.

  13. Direct radioimmunoassay of proinsulin and insulin in human plasma by the chromatographic technique

    Energy Technology Data Exchange (ETDEWEB)

    Megahed, Y M; Abdel-Wahab, M F; El-Shawarbie, K; Sadek, S; Amer, M S [Atomic Energy Establishment, Cairo (Egypt). Radioisotope Department; Ain Shams Univ., Cairo (Egypt). Faculty of Medicine)

    1976-04-01

    Specific method for direct radioimmunoassay of IRP and IRI separately in human plasma has been described. The method is used for extraction of total insulin and separation of IRP from IRI by paper chromatography to be assayed separately. The separation of the two components is identified and confirmed by column chromatography, paper chromatography and ultraviolet spectral analysis in comparison with the standard compounds. 134 plasma samples of different cases were investigated for the determination of IRI, IRP and IRT. Out of these 39 were normals, 16 normal obes, 21 juvinil diabetes, 18 overt adult diabetes, 10 recent adult diabetes, 12 hypothyroidism and 18 bilharzial hepatosplenomegaly. They were used to evaluate the test levels in comparison with blood sugar concentration.

  14. Deciphering a molecular mechanism of neonatal diabetes mellitus by the chemical synthesis of a protein diastereomer, [D-AlaB8]human proinsulin.

    Science.gov (United States)

    Avital-Shmilovici, Michal; Whittaker, Jonathan; Weiss, Michael A; Kent, Stephen B H

    2014-08-22

    Misfolding of proinsulin variants in the pancreatic β-cell, a monogenic cause of permanent neonatal-onset diabetes mellitus, provides a model for a disease of protein toxicity. A hot spot for such clinical mutations is found at position B8, conserved as glycine within the vertebrate insulin superfamily. We set out to investigate the molecular basis of the aberrant properties of a proinsulin clinical mutant in which residue Gly(B8) is replaced by Ser(B8). Modular total chemical synthesis was used to prepare the wild-type [Gly(B8)]proinsulin molecule and three analogs: [D-Ala(B8)]proinsulin, [L-Ala(B8)]proinsulin, and the clinical mutant [L-Ser(B8)]proinsulin. The protein diastereomer [D-Ala(B8)]proinsulin produced higher folding yields at all pH values compared with the wild-type proinsulin and the other two analogs, but showed only very weak binding to the insulin receptor. The clinical mutant [L-Ser(B8)]proinsulin impaired folding at pH 7.5 even in the presence of protein-disulfide isomerase. Surprisingly, although [L-Ser(B8)]proinsulin did not fold well under the physiological conditions investigated, once folded the [L-Ser(B8)]proinsulin protein molecule bound to the insulin receptor more effectively than wild-type proinsulin. Such paradoxical gain of function (not pertinent in vivo due to impaired secretion of the mutant insulin) presumably reflects induced fit in the native mechanism of hormone-receptor engagement. This work provides insight into the molecular mechanism of a clinical mutation in the insulin gene associated with diabetes mellitus. These results dramatically illustrate the power of total protein synthesis, as enabled by modern chemical ligation methods, for the investigation of protein folding and misfolding. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  15. Association of fasting glucagon and proinsulin concentrations with insulin resistance

    DEFF Research Database (Denmark)

    Ferrannini, E; Muscelli, E; Natali, A

    2007-01-01

    AIMS/HYPOTHESIS: Hyperproinsulinaemia and relative hyperglucagonaemia are features of type 2 diabetes. We hypothesised that raised fasting glucagon and proinsulin concentrations may be associated with insulin resistance (IR) in non-diabetic individuals. METHODS: We measured IR [by a euglycaemic......, controlling for known determinants of insulin sensitivity (i.e. sex, age, BMI and glucose tolerance) as well as factors potentially affecting glucagon and proinsulin (i.e. fasting plasma glucose and C-peptide concentrations), glucagon and proinsulin were still positively associated, and adiponectin...

  16. Alternative preparation of inclusion bodies excludes interfering non-protein contaminants and improves the yield of recombinant proinsulin.

    Science.gov (United States)

    Mackin, Robert B

    2014-01-01

    The goal of simple, high-yield expression and purification of recombinant human proinsulin has proven to be a considerable challenge. First, proinsulin forms inclusion bodies during bacterial expression. While this phenomenon can be exploited as a capture step, conventionally prepared inclusion bodies contain significant amounts of non-protein contaminants that interfere with subsequent chromatographic purification. Second, the proinsulin molecules within the inclusion bodies are incorrectly folded, and likely cross-linked to one another, making it difficult to quantify the amount of expressed proinsulin. Third, proinsulin is an intermediate between the initial product of ribosomal translation (preproinsulin) and the final product secreted by pancreatic beta cells (insulin). Therefore, to be efficiently produced in bacteria, it must be produced as an N-terminally extended fusion protein, which has to be converted to authentic proinsulin during the purification scheme. To address all three of these problems, while simultaneously streamlining the procedure and increasing the yield of recombinant proinsulin, we have made three substantive modifications to our previous method for producing proinsulin:.•Conditions for the preparation of inclusion bodies have been altered so contaminants that interfere with semi-preparative reversed-phase chromatography are excluded while the proinsulin fusion protein is retained at high yield.•Aliquots are taken following important steps in the procedure and the quantity of proinsulin-related polypeptide in the sample is compared to the amount present prior to that step.•Final purification is performed using a silica-based reversed-phase matrix in place of a polystyrene-divinylbenzene-based matrix.

  17. Effect of interleukin-1 on the biosynthesis of proinsulin and insulin in isolated rat pancreatic islets

    DEFF Research Database (Denmark)

    Hansen, Birgit Sehested; Linde, S; Spinas, G A

    1988-01-01

    Insulin dependent diabetes mellitus (IDDM) is often preceded or associated with lymphocytic infiltration in the islets of Langerhans (insulitis). We recently demonstrated that interleukin-1 (IL-1) produced by activated macrophages exerts a bimodal effect on insulin release and biosynthesis...... in isolated rat islets. In the present study we have further analysed the effect of recombinant human interleukin-1 beta (rIL-1) on the biosynthesis and conversion of proinsulin 1 and 2 in rat islets. By RP-HPLC-analysis of islets labelled with [3H]leucine we found that exposure to 6 ng/ml of IL-1 for 24 h.......1 to 3.4 +/- 0.4, respectively. Pulse-chase experiments with [3H]leucine and [35S]methionine indicated a more marked reduction in the conversion rate of proinsulin-2 compared to that of proinsulin-1. In conclusion these experiments demonstrate that IL-1 inhibits insulin biosynthesis by preferential...

  18. Mutation in the B chain coding region is associated with impaired proinsulin conversion in a family with hyperproinsulinemia

    International Nuclear Information System (INIS)

    Chan, S.J.; Seino, S.; Gruppuso, P.A.; Schwartz, R.; Steiner, D.F.

    1987-01-01

    A family has recently been described in which hyperproinsulinemia is inherited in an autosomal dominant pattern, suggesting a structural abnormality in the proinsulin molecule as the basis for this disorder. However, unlike two previous kindreds with a similar syndrome, the serum proinsulin-like material in this family did not appear to be an intermediate conversion product but instead behaved like normal human proinsulin by several criteria. To further characterize this disorder the authors isolated and sequenced the insulin gene of the propositus. Leukocyte DNA was cloned in λgt-WES and recombinants containing the two insulin alleles, λMD41 and λMD51, were isolated by plaque hybridization. DNA sequencing of λMD51 showed that it contained the normal coding sequence for human preproinsulin. Sequence analysis of λMD41, however, revealed a single nucleotide substitution in the codon for residue 10 of proinsulin (CAC → GAC) that predicts the exchange of aspartic acid for histidine in the insulin B chain region. This mutation was also found in an insulin allele cloned from a second affected family member (propositus's father). These results strongly implicate this mutation as the cause of the hyperproinsulinemia in this family. Inhibition of the conversion of proinsulin to insulin may be related to altered folding and/or self-association properties of the [Asp 10 ]proinsulin

  19. Low-cost production of proinsulin in tobacco and lettuce chloroplasts for injectable or oral delivery of functional insulin and C-peptide.

    Science.gov (United States)

    Boyhan, Diane; Daniell, Henry

    2011-06-01

    Current treatment for type I diabetes includes delivery of insulin via injection or pump, which is highly invasive and expensive. The production of chloroplast-derived proinsulin should reduce cost and facilitate oral delivery. Therefore, tobacco and lettuce chloroplasts were transformed with the cholera toxin B subunit fused with human proinsulin (A, B, C peptides) containing three furin cleavage sites (CTB-PFx3). Transplastomic lines were confirmed for site-specific integration of transgene and homoplasmy. Old tobacco leaves accumulated proinsulin up to 47% of total leaf protein (TLP). Old lettuce leaves accumulated proinsulin up to 53% TLP. Accumulation was so stable that up to ~40% proinsulin in TLP was observed even in senescent and dried lettuce leaves, facilitating their processing and storage in the field. Based on the yield of only monomers and dimers of proinsulin (3 mg/g leaf, a significant underestimation), with a 50% loss of protein during the purification process, one acre of tobacco could yield up to 20 million daily doses of insulin per year. Proinsulin from tobacco leaves was purified up to 98% using metal affinity chromatography without any His-tag. Furin protease cleaved insulin peptides in vitro. Oral delivery of unprocessed proinsulin bioencapsulated in plant cells or injectable delivery into mice showed reduction in blood glucose levels similar to processed commercial insulin. C-peptide should aid in long-term treatment of diabetic complications including stimulation of nerve and renal functions. Hyper-expression of functional proinsulin and exceptional stability in dehydrated leaves offer a low-cost platform for oral and injectable delivery of cleavable proinsulin. © 2010 The Authors. Plant Biotechnology Journal © 2010 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd.

  20. Analysis of proinsulin and its conversion products by reversed-phase high-performance liquid chromatography

    DEFF Research Database (Denmark)

    Linde, S; Welinder, B S; Nielsen, Jens Høiriis

    1993-01-01

    . Most mammals produce a single insulin, but in rodents two non-allelic insulin genes are expressed. There is an inverse ratio between the two insulins in rats and mice, the reason for this being unknown. It has been suggested that differences in transcription, translation (biosynthesis) and...... PIM (intact proinsulin or its intermediates) has been incompletely determined. Studies of the biosynthesis of proinsulins and their conversion with the purpose of revealing some of these points depend on accessible reversed-phase high-performance liquid chromatographic (RP-HPLC) analyses capable...

  1. Kinetics of circulating endogenous insulin, C-peptide, and proinsulin in fasting nondiabetic man

    DEFF Research Database (Denmark)

    Henriksen, Jens Henrik Sahl; Tronier, B; Bülow, J B

    1987-01-01

    Plasma concentrations of insulin, C-peptide, and proinsulin were measured in different vascular beds in order to determine renal, hepatic, and systemic kinetics of the endogenous peptides in the fasting condition. Nineteen nondiabetic subjects were studied, two were normal, nine had minor vascular...

  2. Specific insulin and proinsulin secretion in glucokinase-deficient individuals

    Directory of Open Access Journals (Sweden)

    V.C. Pardini

    1999-04-01

    Full Text Available Glucokinase (GCK is an enzyme that regulates insulin secretion, keeping glucose levels within a narrow range. Mutations in the glucokinase gene cause a rare form of diabetes called maturity-onset diabetes of the young (MODY. An early onset (less than 25 years, autosomal dominant inheritance and low insulin secretion stimulated by glucose characterize MODY patients. Specific insulin and proinsulin were measured in serum by immunofluorimetric assays (IFMA during a 75-g oral glucose tolerance test (OGTT. Two kindreds (SA and LZ were studied and compared to non-diabetic unrelated individuals (control group 1 matched for age and body mass index (BMI. In one kindred, some of these subjects were also obese (BMI >26 kg/m2, and other family members also presented with obesity and/or late-onset NIDDM. The MODY patients were also compared to a group of five of their first-degree relatives with obesity and/or late-onset NIDDM. The proinsulin profile was different in members of the two MODY kindreds. Fasting proinsulin and the proinsulin/insulin ratio were similar in MODY members of kindred LZ and subjects from control group 1, but were significantly lower than in MODY members of kindred SA (P<0.02 and P<0.01, for proinsulin and proinsulin/insulin ratio, respectively. Moreover, MODY members of family SA had higher levels of proinsulin and proinsulin/insulin ratio, although not significantly different, when compared to their first-degree relatives and to subjects from control group 2. In conclusion, we observed variable degrees of proinsulin levels and proinsulin/insulin ratio in MODY members of two different kindreds. The higher values of these parameters found in MODY and non-MODY members of kindred SA is probably related to the obesity and late-onset NIDDM background present in this family.

  3. Proinsulin, adiponectin and hsCRP in reproductive age women with polycystic ovary syndrome (PCOS)--the effect of metformin treatment.

    Science.gov (United States)

    Kruszyńska, Aleksandra; Słowińska-Srzednicka, Jadwiga; Jeske, Wojciech; Zgliczyński, Wojciech

    2014-01-01

    Women with polycystic ovary syndrome (PCOS) often suffer from obesity and insulin resistance. The role of proinsulin, which is known to be an indicator of fertility outcomes in PCOS women, and that of adiponectin, in the pathogenesis of PCOS is not well elucidated. Our objective was to determine proinsulin, adiponectin, hsCRP and other hormonal and metabolic parameters in PCOS women before and after metformin treatment. Two PCOS groups of patients of reproductive age (90 lean and 88 obese or overweight) with two control groups, adjusted for body mass index (BMI), were compared at baseline. 32 PCOS women were studied at baseline, after three and six months of metformin (1,000 mg/day) treatment. Clinical, anthropometric, biochemical and hormonal parameters were assessed. Proinsulin and hsCRP levels were the highest in obese PCOS women and were statistically different than in lean PCOS women (proinsulin: 11.4 v. 6.9 pmol/L; hsCRP 2.46 v. 0.47 mg/L, p treatment only in obese PCOS women. PCOS, when accompanied by obesity, is associated with elevated proinsulin concentrations, which correlates with higher hsCRP and increased FAI. Proinsulin level decreases due to metformin treatment. Our results suggest that obese or overweight PCOS and lean PCOS are characterised by different hormonal and metabolic parameters and have a different response to metformin treatment.

  4. The PTPN22 C1858T gene variant is associated with proinsulin in new-onset type 1 diabetes

    Directory of Open Access Journals (Sweden)

    Vanelli Maurizio

    2011-03-01

    Full Text Available Abstract Background The protein tyrosine phosphatase nonreceptor type 2 (PTPN22 has been established as a type 1 diabetes susceptibility gene. A recent study found the C1858T variant of this gene to be associated with lower residual fasting C-peptide levels and poorer glycemic control in patients with type 1 diabetes. We investigated the association of the C1858T variant with residual beta-cell function (as assessed by stimulated C-peptide, proinsulin and insulin dose-adjusted HbA1c, glycemic control, daily insulin requirements, diabetic ketoacidosis (DKA and diabetes-related autoantibodies (IA-2A, GADA, ICA, ZnT8Ab in children during the first year after diagnosis of type 1 diabetes. Methods The C1858T variant was genotyped in an international cohort of children (n = 257 patients with newly diagnosed type 1 diabetes during 12 months after onset. We investigated the association of this variant with liquid-meal stimulated beta-cell function (proinsulin and C-peptide and antibody status 1, 6 and 12 months after onset. In addition HbA1c and daily insulin requirements were determined 1, 3, 6, 9 and 12 months after diagnosis. DKA was defined at disease onset. Results A repeated measurement model of all time points showed the stimulated proinsulin level is significantly higher (22%, p = 0.03 for the T allele carriers the first year after onset. We also found a significant positive association between proinsulin and IA levels (est.: 1.12, p = 0.002, which did not influence the association between PTPN22 and proinsulin (est.: 1.28, p = 0.03. Conclusions The T allele of the C1858T variant is positively associated with proinsulin levels during the first 12 months in newly diagnosed type 1 diabetes children.

  5. Studies on the regioselectivity and kinetics of the action of trypsin on proinsulin and its derivatives using mass spectrometry.

    Science.gov (United States)

    Gardner, Qurra-tul-Ann Afza; Younas, Hooria; Akhtar, Muhammad

    2013-01-01

    Human M-proinsulin was cleaved by trypsin at the R(31)R(32)-E(33) and K(64)R(65)-G(66) bonds (B/C and C/A junctions), showing the same cleavage specificity as exhibited by prohormone convertases 1 and 2 respectively. Buffalo/bovine M-proinsulin was also cleaved by trypsin at the K(59)R(60)-G(61) bond but at the B/C junction cleavage occurred at the R(31)R(32)-E(33) as well as the R(31)-R(32)E(33) bond. Thus, the human isoform in the native state, with a 31 residue connecting C-peptide, seems to have a unique structure around the B/C and C/A junctions and cleavage at these sites is predominantly governed by the structure of the proinsulin itself. In the case of both the proinsulin species the cleavage at the B/C junction was preferred (65%) over that at the C/A junction (35%) supporting the earlier suggestion of the presence of some form of secondary structure at the C/A junction. Proinsulin and its derivatives, as natural substrates for trypsin, were used and mass spectrometric analysis showed that the k(cat.)/K(m) values for the cleavage were most favourable for the scission of the bonds at the two junctions (1.02±0.08×10(5)s(-1)M(-1)) and the cleavage of the K(29)-T(30) bond of M-insulin-RR (1.3±0.07×10(5)s(-1)M(-1)). However, the K(29)-T(30) bond in M-insulin, insulin as well as M-proinsulin was shielded from attack by trypsin (k(cat.)/K(m) values around 1000s(-1)M(-1)). Hence, as the biosynthetic path follows the sequence; proinsulin→insulin-RR→insulin, the K(29)-T(30) bond becomes shielded, exposed then shielded again respectively. Copyright © 2012 Elsevier B.V. All rights reserved.

  6. Release of immunoreactive and radioactively prelabelled endogenous (pro-)insulin from isolated islets of rat pancreas in the presence of exogenous insulin

    Energy Technology Data Exchange (ETDEWEB)

    Schatz, H [Giessen Univ. (Germany, F.R.). Zentrum fuer Innere Medizin; Pfeiffer, E F

    1977-01-01

    To study the influence of insulin on its secretion, collagenase-isolated islets of rat pancreas were prelabelled with (/sup 3/H)leucine for 2 h. After washing the islets, (pro-)insulin release was stimulated by glucose in the presence or absence of exogenous insulin (up to 2.5 mu./ml. Hormone release was unchanged by the presence of exogenous insulin as judged by determination of both immunoreactive insulin and radioactivity incorporated into the proinsulin and insulin fractions of the medium. No direct feedback mechanism for insulin secretion was apparent from this study.

  7. Increased seroreactivity to proinsulin and homologous mycobacterial peptides in latent autoimmune diabetes in adults.

    Directory of Open Access Journals (Sweden)

    Magdalena Niegowska

    Full Text Available Latent Autoimmune Diabetes in Adults (LADA is a slowly progressing form of immune-mediated diabetes that combines phenotypical features of type 2 diabetes (T2D with the presence of islet cell antigens detected in type 1 diabetes (T1D. Heterogeneous clinical picture have led to the classification of patients based on the levels of antibodies against glutamic acid decarboxylase 65 (GADA that correlate with clinical phenotypes closer to T1D or T2D when GADA titers are high or low, respectively. To date, LADA etiology remains elusive despite numerous studies investigating on genetic predisposition and environmental risk factors. To our knowledge, this is the first study aimed at evaluation of a putative role played by Mycobacterium avium subsp. paratuberculosis (MAP as an infective agent in LADA pathogenesis. MAP is known to cause chronic enteritis in ruminants and has been associated with autoimmune disorders in humans. We analyzed seroreactivity of 223 Sardinian LADA subjects and 182 healthy volunteers against MAP-derived peptides and their human homologs of proinsulin and zinc transporter 8 protein. A significantly elevated positivity for MAP/proinsulin was detected among patients, with the highest prevalence in the 32-41-year-old T1D-like LADA subgroup, supporting our hypothesis of a possible MAP contribution in the development of autoimmunity.

  8.  Pleiotropic action of proinsulin C-peptid

    Directory of Open Access Journals (Sweden)

    Michał Usarek

    2012-03-01

    Full Text Available  Proinsulin C-peptide, released in equimolar amounts with insulin by pancreatic β cells, since its discovery in 1967 has been thought to be devoid of biological functions apart from correct insulin processing and formation of disulfide bonds between A and B chains. However, in the last two decades research has brought a substantial amount of data indicating a crucial role of C-peptide in regulating various processes in different types of cells and organs. C-peptide acts presumably via either G-protein-coupled receptor or directly inside the cell, after being internalized. However, a receptor binding this peptide has not been identified yet. This peptide ameliorates pathological changes induced by type 1 diabetes mellitus, including glomerular hyperfiltration, vessel endothelium inflammation and neuron demyelinization. In diabetic patients and diabetic animal models, C-peptide substitution in physiological doses improves the functional and structural properties of peripheral neurons and protects against hyperglycemia-induced apoptosis, promoting neuronal development, regeneration and cell survival. Moreover, it affects glycogen synthesis in skeletal muscles. In vitro C-peptide promotes disaggregation of insulin oligomers, thus enhancing its bioavailability and effects on metabolism. There are controversies concerning the biological action of C-peptide, particularly with respect to its effect on Na /K -ATPase activity. Surprisingly, the excess of circulating peptide associated with diabetes type 2 contributes to atherosclerosis development. In view of these observations, long-term, large-scale clinical investigations using C-peptide physiological doses need to be conducted in order to determine safety and health outcomes of long-term administration of C-peptide to diabetic patients.

  9. Comparative assessment of immunization procedures for development of anti proinsulin antisera for radioimmunoassay

    International Nuclear Information System (INIS)

    Nascimento, M. do; Borghi, V.C.; Bellini, M.H.; Mesquita, C.H.; Wajchenberg, B.L.

    1992-08-01

    Two schedules of immunization were employed for developing anti proinsulin antisera for radioimmunoassay. Biosynthetic human proinsulin-h P I (Elli Lilly. US), was injected subcutaneously in guinea pigs in multiple sites. In the first schedule were used 50 u g of h P I and the booster injections were administered 4 weeks after the primary immunization and then at 3-week intervals. In the second schedule was used 250 u g of h P I and boosters were done 7, 9 and 18 weeks later. As the antisera were not sufficiently specific for h P I they were purified and assessed for kinetic of precipitation and avidity. Both immunization schedules gave comparable responses. The antisera generated by the use of 50 u g of h P I presented higher cross-reactivity with insulin while the reactivity with c p eptide was of the same order in both antiserum groups. The avidity was very variable in the two groups and the three most sensitive antisera required 24 h at 4 o C for achieving maximum binding with the 125 I-h P I. However, only one antiserum (from the first group) was suitable for the radioimmunoassay. This study emphasizes the difficulties of making valid comparisons between different immunization procedures, especially in the cases when highest avidity is required. (author)

  10. Immunological half-life of porcine proinsulin C-peptide

    Energy Technology Data Exchange (ETDEWEB)

    Oyama, H; Horino, M; Matsumura, S [Kawasaki Medical Coll., Kurashiki (Japan). Div. of Endocrinology; Kobayshi, K; Suetsugu, N [Yamaguchi Univ., Ube (Japan). School of Medicine

    1975-11-01

    Immunological half-lifes of injected porcine C-peptide and insulin with RIA were studied and calculated as 9.8 and 8.0 minutes. Higher circulating levels of C-peptide as compared to insulin in normal young swines lead to speculation about a longer half-life of C-peptide. This hypothesis was verified in this study. Immunological half-lifes of porcine proinsulin and insulin in the pig were 20 and 6 minutes, respectively.

  11. Fasting proinsulin levels are significantly associated with 20 year cancer mortality rates. The Hoorn Study

    NARCIS (Netherlands)

    Walraven, I.; van 't Riet, E.; Stehouwer, C.D.A.; Polak, B.C.P.; Moll, A.C.; Dekker, J.M.; Nijpels, G.

    2013-01-01

    Aims/hypothesis: Proinsulin is possibly associated with cancer through activation of insulin receptor isoform A. We sought to investigate the associations between proinsulin and 20 year cancer mortality rates. Methods: The study was performed within the Hoorn Study, a population-based study of

  12. Quantitative variation in obesity-related traits and insulin precursors linked to the OB gene region on human chromosome 7

    Energy Technology Data Exchange (ETDEWEB)

    Duggirala, R.; Stern, M.P.; Reinhart, L.J. [Univ. of Texas Health Science Center, San Antonio, TX (United States)] [and others

    1996-09-01

    Despite the evidence that human obesity has strong genetic determinants, efforts at identifying specific genes that influence human obesity have largely been unsuccessful. Using the sibship data obtained from 32 low-income Mexican American pedigrees ascertained on a type II diabetic proband and a multipoint variance-components method, we tested for linkage between various obesity-related traits plus associated metabolic traits and 15 markers on human chromosome 7. We found evidence for linkage between markers in the OB gene region and various traits, as follows: D7S514 and extremity skinfolds (LOD = 3.1), human carboxypeptidase A1 (HCPA1) and 32,33-split proinsulin level (LOD = 4.2), and HCPA1 and proinsulin level (LOD = 3.2). A putative susceptibility locus linked to the marker D7S514 explained 56% of the total phenotypic variation in extremity skinfolds. Variation at the HCPA1 locus explained 64% of phenotypic variation in proinsulin level and {approximately}73% of phenotypic variation in split proinsulin concentration, respectively. Weaker evidence for linkage to several other obesity-related traits (e.g., waist circumference, body-mass index, fat mass by bioimpedance, etc.) was observed for a genetic location, which is {approximately}15 cM telomeric to OB. In conclusion, our study reveals that the OB region plays a significant role in determining the phenotypic variation of both insulin precursors and obesity-related traits, at least in Mexican Americans. 66 refs., 3 figs., 4 tabs.

  13. Mutant INS-gene induced diabetes of youth: proinsulin cysteine residues impose dominant-negative inhibition on wild-type proinsulin transport.

    Directory of Open Access Journals (Sweden)

    Ming Liu

    2010-10-01

    Full Text Available Recently, a syndrome of Mutant INS-gene-induced Diabetes of Youth (MIDY, derived from one of 26 distinct mutations has been identified as a cause of insulin-deficient diabetes, resulting from expression of a misfolded mutant proinsulin protein in the endoplasmic reticulum (ER of insulin-producing pancreatic beta cells. Genetic deletion of one, two, or even three alleles encoding insulin in mice does not necessarily lead to diabetes. Yet MIDY patients are INS-gene heterozygotes; inheritance of even one MIDY allele, causes diabetes. Although a favored explanation for the onset of diabetes is that insurmountable ER stress and ER stress response from the mutant proinsulin causes a net loss of beta cells, in this report we present three surprising and interlinked discoveries. First, in the presence of MIDY mutants, an increased fraction of wild-type proinsulin becomes recruited into nonnative disulfide-linked protein complexes. Second, regardless of whether MIDY mutations result in the loss, or creation, of an extra unpaired cysteine within proinsulin, Cys residues in the mutant protein are nevertheless essential in causing intracellular entrapment of co-expressed wild-type proinsulin, blocking insulin production. Third, while each of the MIDY mutants induces ER stress and ER stress response; ER stress and ER stress response alone appear insufficient to account for blockade of wild-type proinsulin. While there is general agreement that ultimately, as diabetes progresses, a significant loss of beta cell mass occurs, the early events described herein precede cell death and loss of beta cell mass. We conclude that the molecular pathogenesis of MIDY is initiated by perturbation of the disulfide-coupled folding pathway of wild-type proinsulin.

  14. 21 CFR 862.1135 - C-peptides of proinsulin test system.

    Science.gov (United States)

    2010-04-01

    .... Measurements of C-peptides of proinsulin are used in the diagnosis and treatment of patients with abnormal insulin secretion, including diabetes mellitus. (b) Classification. Class I (general controls). The device...

  15. Mesenchymal Stem Cell Differentiation into Adipocytes Is Equally Induced by Insulin and Proinsulin In Vitro.

    Science.gov (United States)

    Pfützner, Andreas; Schipper, Dorothee; Pansky, Andreas; Kleinfeld, Claudia; Roitzheim, Barbara; Tobiasch, Edda

    2017-11-30

    In advanced β -cell dysfunction, proinsulin is increasingly replacing insulin as major component of the secretion product. It has been speculated that proinsulin has at least the same adipogenic potency than insulin, leading to an increased tendency of lipid tissue formation in patients with late stage β -cell dysfunction. Mesenchymal stem cells obtained from liposuction material were grown in differentiation media containing insulin (0.01 μmol), proinsulin (0.01 μmol) or insulin+proinsulin (each 0.005 μmol). Cell culture supernatants were taken from these experiments and an untreated control at weeks 1, 2, and 3, and were stored at -80°C until analysis. Cell differentiation was microscopically supervised and adiponectin concentrations were measured as marker for differentiation into mature lipid cells. This experiment was repeated three times. No growth of lipid cells and no change in adiponectin values was observed in the negative control group (after 7/14/12 days: 3.2±0.5/3.3±0.1/4.4±0.5 ng/ml/12 h). A continuous differentiation into mature adipocytes (also confirmed by Red-Oil-staining) and a corresponding increase in adiponectin values was observed in the experiments with insulin (3.6±1.9/5.1±1.4/13.3±1.5 ng/ml/12 h; p<0.05 week 1 vs. week 3) and proinsulin (3.3±1.2/3.5±0.3/12.2±1.2 ng/ml/12 h; p<0.05). Comparable effects were seen with the insulin/proinsulin combination. Proinsulin has the same adipogenic potential than insulin in vitro. Proinsulin has only 10∼20% of the glucose-lowering effect of insulin. It can be speculated that the adipogenic potential of proinsulin may be a large contributor to the increased body weight problems in patients with type 2 diabetes and advanced β -cell dysfunction.

  16. Proinsulin-expressing dendritic cells in type 2 neuropathic diabetic patients with and without foot lesions.

    Science.gov (United States)

    Sambataro, Maria; Sambado, Luisa; Trevisiol, Enrica; Cacciatore, Matilde; Furlan, Anna; Stefani, Piero Maria; Seganfreddo, Elena; Durante, Elisabetta; Conte, Stefania; Della Bella, Silvia; Paccagnella, Agostino; Dei Tos, Angelo Paolo

    2018-02-12

    Diabetic neuropathy is the most common complication of diabetes and is frequently associated with foot ischemia and infection, but its pathogenesis is controversial. We hypothesized that proinsulin expression in peripheral blood mononuclear cells is a process relevant to this condition and could represent a link among hyperglycemia, nerve susceptibility, and diabetic foot lesions. We assessed proinsulin expression by using flow cytometry in dendritic cells from control participants and patients with type 2 diabetic with or without peripheral neuropathy or accompanied by diabetic foot. Among 32 non-neuropathic and 120 neuropathic patients with type 2 diabetic, we performed leg electromyography and found average sensory sural nerve conduction velocities of 48 ± 4 and 30 ± 4 m/s, respectively ( P foot lesions, and 39 had neuroischemic foot lesions (allux oximetry diabetic population, but not in nondiabetic participants, a progressively increasing level of peripheral blood dendritic cell proinsulin expression was detected, which directly correlated with circulating TNF-α levels ( P diabetes, proinsulin-expressing blood cells, possibly via their involvement in innate immunity, may play a role in diabetic peripheral neuropathy and foot lesions.-Sambataro, M., Sambado, L., Trevisiol, E., Cacciatore, M., Furlan, A., Stefani, P. M., Seganfreddo, E., Durante, E., Conte, S., Della Bella, S., Paccagnella, A., dei Tos, A. P. Proinsulin-expressing dendritic cells in type 2 neuropathic diabetic patients with and without foot lesions.

  17. Proinsulin C-peptide interferes with insulin fibril formation

    International Nuclear Information System (INIS)

    Landreh, Michael; Stukenborg, Jan-Bernd; Willander, Hanna; Söder, Olle; Johansson, Jan; Jörnvall, Hans

    2012-01-01

    Highlights: ► Insulin and C-peptide can interact under insulin fibril forming conditions. ► C-peptide is incorporated into insulin aggregates and alters aggregation lag time. ► C-peptide changes insulin fibril morphology and affects backbone accessibility. ► C-peptide may be a regulator of fibril formation by β-cell granule proteins. -- Abstract: Insulin aggregation can prevent rapid insulin uptake and cause localized amyloidosis in the treatment of type-1 diabetes. In this study, we investigated the effect of C-peptide, the 31-residue peptide cleaved from proinsulin, on insulin fibrillation at optimal conditions for fibrillation. This is at low pH and high concentration, when the fibrils formed are regular and extended. We report that C-peptide then modulates the insulin aggregation lag time and profoundly changes the fibril appearance, to rounded clumps of short fibrils, which, however, still are Thioflavine T-positive. Electrospray ionization mass spectrometry also indicates that C-peptide interacts with aggregating insulin and is incorporated into the aggregates. Hydrogen/deuterium exchange mass spectrometry further reveals reduced backbone accessibility in insulin aggregates formed in the presence of C-peptide. Combined, these effects are similar to those of C-peptide on islet amyloid polypeptide fibrillation and suggest that C-peptide has a general ability to interact with amyloidogenic proteins from pancreatic β-cell granules. Considering the concentrations, these peptide interactions should be relevant also during physiological secretion, and even so at special sites post-secretory or under insulin treatment conditions in vivo.

  18. Proinsulin C-peptide interferes with insulin fibril formation

    Energy Technology Data Exchange (ETDEWEB)

    Landreh, Michael [Department of Medical Biochemistry and Biophysics, Karolinska Institutet, S-171 77 Stockholm (Sweden); Stukenborg, Jan-Bernd [Department of Women' s and Children' s Health, Astrid Lindgren Children' s Hospital, Pediatric Endocrinology Unit, Karolinska Institutet and University Hospital, S-17176 Stockholm (Sweden); Willander, Hanna [KI-Alzheimer' s Disease Research Center, NVS Department, Karolinska Institutet, S-141 86 Stockholm (Sweden); Soeder, Olle [Department of Women' s and Children' s Health, Astrid Lindgren Children' s Hospital, Pediatric Endocrinology Unit, Karolinska Institutet and University Hospital, S-17176 Stockholm (Sweden); Johansson, Jan [KI-Alzheimer' s Disease Research Center, NVS Department, Karolinska Institutet, S-141 86 Stockholm (Sweden); Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, S-751 23 Uppsala (Sweden); Joernvall, Hans, E-mail: Hans.Jornvall@ki.se [Department of Medical Biochemistry and Biophysics, Karolinska Institutet, S-171 77 Stockholm (Sweden)

    2012-02-17

    Highlights: Black-Right-Pointing-Pointer Insulin and C-peptide can interact under insulin fibril forming conditions. Black-Right-Pointing-Pointer C-peptide is incorporated into insulin aggregates and alters aggregation lag time. Black-Right-Pointing-Pointer C-peptide changes insulin fibril morphology and affects backbone accessibility. Black-Right-Pointing-Pointer C-peptide may be a regulator of fibril formation by {beta}-cell granule proteins. -- Abstract: Insulin aggregation can prevent rapid insulin uptake and cause localized amyloidosis in the treatment of type-1 diabetes. In this study, we investigated the effect of C-peptide, the 31-residue peptide cleaved from proinsulin, on insulin fibrillation at optimal conditions for fibrillation. This is at low pH and high concentration, when the fibrils formed are regular and extended. We report that C-peptide then modulates the insulin aggregation lag time and profoundly changes the fibril appearance, to rounded clumps of short fibrils, which, however, still are Thioflavine T-positive. Electrospray ionization mass spectrometry also indicates that C-peptide interacts with aggregating insulin and is incorporated into the aggregates. Hydrogen/deuterium exchange mass spectrometry further reveals reduced backbone accessibility in insulin aggregates formed in the presence of C-peptide. Combined, these effects are similar to those of C-peptide on islet amyloid polypeptide fibrillation and suggest that C-peptide has a general ability to interact with amyloidogenic proteins from pancreatic {beta}-cell granules. Considering the concentrations, these peptide interactions should be relevant also during physiological secretion, and even so at special sites post-secretory or under insulin treatment conditions in vivo.

  19. Prediction of Impending Type 1 Diabetes through Automated Dual-Label Measurement of Proinsulin:C-Peptide Ratio.

    Directory of Open Access Journals (Sweden)

    Annelien Van Dalem

    Full Text Available The hyperglycemic clamp test, the gold standard of beta cell function, predicts impending type 1 diabetes in islet autoantibody-positive individuals, but the latter may benefit from less invasive function tests such as the proinsulin:C-peptide ratio (PI:C. The present study aims to optimize precision of PI:C measurements by automating a dual-label trefoil-type time-resolved fluorescence immunoassay (TT-TRFIA, and to compare its diagnostic performance for predicting type 1 diabetes with that of clamp-derived C-peptide release.Between-day imprecision (n = 20 and split-sample analysis (n = 95 were used to compare TT-TRFIA (AutoDelfia, Perkin-Elmer with separate methods for proinsulin (in-house TRFIA and C-peptide (Elecsys, Roche. High-risk multiple autoantibody-positive first-degree relatives (n = 49; age 5-39 were tested for fasting PI:C, HOMA2-IR and hyperglycemic clamp and followed for 20-57 months (interquartile range.TT-TRFIA values for proinsulin, C-peptide and PI:C correlated significantly (r2 = 0.96-0.99; P<0.001 with results obtained with separate methods. TT-TRFIA achieved better between-day %CV for PI:C at three different levels (4.5-7.1 vs 6.7-9.5 for separate methods. In high-risk relatives fasting PI:C was significantly and inversely correlated (rs = -0.596; P<0.001 with first-phase C-peptide release during clamp (also with second phase release, only available for age 12-39 years; n = 31, but only after normalization for HOMA2-IR. In ROC- and Cox regression analysis, HOMA2-IR-corrected PI:C predicted 2-year progression to diabetes equally well as clamp-derived C-peptide release.The reproducibility of PI:C benefits from the automated simultaneous determination of both hormones. HOMA2-IR-corrected PI:C may serve as a minimally invasive alternative to the more tedious hyperglycemic clamp test.

  20. Expression of cholera toxin B–proinsulin fusion protein in lettuce and tobacco chloroplasts – oral administration protects against development of insulitis in non-obese diabetic mice

    OpenAIRE

    Ruhlman, Tracey; Ahangari, Raheleh; Devine, Andrew; Samsam, Mohtahsem; Daniell, Henry

    2007-01-01

    Lettuce and tobacco chloroplast transgenic lines expressing the cholera toxin B subunit–human proinsulin (CTB-Pins) fusion protein were generated. CTB-Pins accumulated up to ~16% of total soluble protein (TSP) in tobacco and up to ~2.5% of TSP in lettuce. Eight milligrams of powdered tobacco leaf material expressing CTB-Pins or, as negative controls, CTB–green fluorescent protein (CTB-GFP) or interferon–GFP (IFN-GFP), or untransformed leaf, were administered orally, each week for 7 weeks, to ...

  1. Heterogeneity of human plasma insulin: techniques for separating immunoreactive components and their determination by radioimmunoassay

    International Nuclear Information System (INIS)

    Souza, Iracelia Torres de Toledo e

    1977-01-01

    When human plasma is filtered on Sephadex G-SO fine, insulin immunoreactivity is recovered in two peaks: 'big insulin', the higher molecular weight component and 'little insulin', the lower molecular component, having elution volumes that correspond to those of porcine proinsulin 125 I and porcine insulin 125 I respectively. The presence of another form of immunoreactive insulin 'big big insulin' was detected from an insuloma suspect and its elution pattern corresponding to serum albumin. The eluates correspondent to 'big' and 'little' insulin as well as 'big big' component were assayed by radioimmunoassay using crystalline human insulin as a standard, porcine insulin 125 tracer and anti insulin serum. The antibody, raised in guinea-pigs, was sensitive and potent being adequate for the assay. The reactivity of insulin and proinsulin was tested against the antibody. The relative proportions of several components of total immunoreactive insulin in plasma were studied in basal conditions in five normal subjects and in the patient JSC with pancreatic insulin-secreting tumor as well as after glucose stimuli in all tolbutamide in JSC. (author)

  2. Proinsulin is stable at room temperature for 24 hours in EDTA: A clinical laboratory analysis (adAPT 3.

    Directory of Open Access Journals (Sweden)

    Jane Davidson

    Full Text Available Reference laboratories advise immediate separation and freezing of samples for the assay of proinsulin, which limit its practicability for smaller centres. Following the demonstration that insulin and C-peptide are stable in EDTA at room temperature for at least 24hours, we undertook simple stability studies to establish whether the same might apply to proinsulin.Venous blood samples were drawn from six adult women, some fasting, some not, aliquoted and assayed immediately and after storage at either 4°C or ambient temperature for periods from 2h to 24h.There was no significant variation or difference with storage time or storage condition in either individual or group analysis.Proinsulin appears to be stable at room temperature in EDTA for at least 24h. Immediate separation and storage on ice of samples for proinsulin assay is not necessary, which will simplify sample transport, particularly for multicentre trials.

  3. Proinsulin maturation disorder is a contributor to the defect of subsequent conversion to insulin in {beta}-cells

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Jie, E-mail: jie.wang2@osumc.edu [Division of Endocrinology, Diabetes and Metabolism, Department of Internal Medicine, The Ohio State University, Columbus, OH (United States); Osei, Kwame [Division of Endocrinology, Diabetes and Metabolism, Department of Internal Medicine, The Ohio State University, Columbus, OH (United States)

    2011-07-22

    Highlights: {yields} Primary proinsulin maturation disorder is inherent in Ins2{sup +/Akita} islets/{beta}-cells. {yields} A consequence is the inefficient conversion of proinsulin to insulin. {yields} Post-translational defects occur as well in the involved PC1/3 and PC2 convertases. {yields} Proinsulin maturation chaos results in defects in the following conversion process. {yields} A link of the proinsulin maturation disorder and hyperproinsulinemia is suggested. -- Abstract: Disproportionate hyperproinsulinemia is an indicator of {beta}-cell dysfunction in diabetes and the basis underlying this abnormality remains obscure. Recently, we have found proinsulin is an aggregation-prone molecule inherent with a low relative folding rate and maintains a homeostatic balance of natively and plentiful non-natively folded states (i.e., proinsulin homeostasis, PIHO) in normal {beta}-cells as a result of the integration of maturation and disposal processes. PIHO is susceptible to environmental and genetic influences. Perturbation of PIHO produces a number of toxic consequences with known association to {beta}-cell failure in diabetes. To explore whether the perturbation of PIHO has a link to disproportionate hyperproinsulinemia, we investigated proinsulin conversion and the involved prohormone convertase 1/3 (PC1/3) and 2 (PC2) in mouse Ins2{sup +/Akita} islets/{beta}-cells that preserve a primary PIHO disorder due to a mutation (C96Y) in the insulin 2 (Ins2) gene. Our metabolic-labeling studies found an increased ratio of proinsulin to insulin in the cellular or released proteins of Ins2{sup +/Akita} islets. Histological, metabolic-labeling, and RT-PCR analyses revealed decreases of the PC1/3 and PC2 immunoreactivities in the {beta}-cells of Ins2{sup +/Akita} islets in spite of no declines of these two convertases at the transcriptional and translational levels. Immunoblot analyses in cloned Ins2{sup +/Akita} {beta}-cells further confirmed the increased ratio of proinsulin

  4. Proinsulin maturation disorder is a contributor to the defect of subsequent conversion to insulin in β-cells

    International Nuclear Information System (INIS)

    Wang, Jie; Osei, Kwame

    2011-01-01

    Highlights: → Primary proinsulin maturation disorder is inherent in Ins2 +/Akita islets/β-cells. → A consequence is the inefficient conversion of proinsulin to insulin. → Post-translational defects occur as well in the involved PC1/3 and PC2 convertases. → Proinsulin maturation chaos results in defects in the following conversion process. → A link of the proinsulin maturation disorder and hyperproinsulinemia is suggested. -- Abstract: Disproportionate hyperproinsulinemia is an indicator of β-cell dysfunction in diabetes and the basis underlying this abnormality remains obscure. Recently, we have found proinsulin is an aggregation-prone molecule inherent with a low relative folding rate and maintains a homeostatic balance of natively and plentiful non-natively folded states (i.e., proinsulin homeostasis, PIHO) in normal β-cells as a result of the integration of maturation and disposal processes. PIHO is susceptible to environmental and genetic influences. Perturbation of PIHO produces a number of toxic consequences with known association to β-cell failure in diabetes. To explore whether the perturbation of PIHO has a link to disproportionate hyperproinsulinemia, we investigated proinsulin conversion and the involved prohormone convertase 1/3 (PC1/3) and 2 (PC2) in mouse Ins2 +/Akita islets/β-cells that preserve a primary PIHO disorder due to a mutation (C96Y) in the insulin 2 (Ins2) gene. Our metabolic-labeling studies found an increased ratio of proinsulin to insulin in the cellular or released proteins of Ins2 +/Akita islets. Histological, metabolic-labeling, and RT-PCR analyses revealed decreases of the PC1/3 and PC2 immunoreactivities in the β-cells of Ins2 +/Akita islets in spite of no declines of these two convertases at the transcriptional and translational levels. Immunoblot analyses in cloned Ins2 +/Akita β-cells further confirmed the increased ratio of proinsulin to insulin despite the levels of PC1/3 and PC2 proteins were not reduced

  5. Insulin secretion and glucose uptake by isolated islets of the hamster. Effect of insulin, proinsulin and C-peptide

    Energy Technology Data Exchange (ETDEWEB)

    Dunbar, J C; McLaughlin, W J; Walsh, M F.J.; Foa, P P [Sinai Hospital of Detroit, Mich. (USA). Dept. of Research

    1976-01-01

    Isolated pancreatic islets of normal hamsters were perfused either in a closed or in a open system. When the buffer was recirculated and the endogenous insulin was allowed to accumulate, the islets secreted significantly less insulin than when the system was open and the endogenous insulin was washed away. The addition of monocomponent insulin or of proinsulin to the perfusion buffer significantly decreased insulin secretion. The inhibitory action of proinsulin was significantly greater than that of monocomponent insulin. C peptide had no effect. When pancreatic islets were incubated in a fixed volume of stationary buffer containing unlabeled glucose (1.0 mg or 3.0 mg/ml) and glucose-U-/sup 14/C (1.0 ..mu..C/ml), the amount of insulin secreted and the /sup 14/CO/sub 2/ produced by each islet decreased progressively as the number of islets in the sample increased. Under these conditions, the concentration of insulin required to inhibit insulin secretion increased with the concentration of glucose in the medium. Proinsulin did not alter the incorporation of leucine-4.5-/sup 3/H into total extractable insulin (insulin + proinsulin). Thus, insulin and proinsulin appear to inhibit insulin release, but not insulin synthesis.

  6. Determination of Human Hemoglobin Derivatives.

    Science.gov (United States)

    Attia, Atef M M; Ibrahim, Fatma A A; Abd El-Latif, Noha A; Aziz, Samir W; Abdelmottaleb Moussa, Sherif A; Elalfy, Mohsen S

    2015-01-01

    The levels of the inactive hemoglobin (Hb) pigments [such as methemoglobin (metHb), carboxyhemoglobin (HbCO) and sulfohemoglobin (SHb)] and the active Hb [in the oxyhemoglobin (oxyHb) form] as well as the blood Hb concentration in healthy non pregnant female volunteers were determined using a newly developed multi-component spectrophotometric method. The results of this method revealed values of SHb% in the range (0.0727-0.370%), metHb% (0.43-1.0%), HbCO% (0.4-1.52%) and oxyHb% (97.06-98.62%). Furthermore, the results of this method revealed values of blood Hb concentration in the range (12.608-15.777 g/dL). The method is highly sensitive, accurate and reproducible.

  7. Genome-wide association identifies nine common variants associated with fasting proinsulin levels and provides new insights into the pathophysiology of type 2 diabetes

    NARCIS (Netherlands)

    R.J. Strawbridge (Rona); J. Dupuis (Josée); I. Prokopenko (Inga); A.M. Barker (Adam); E. Ahlqvist (Emma); D. Rybin (Denis); J.R. Petrie (John); N. Bouatia-Naji (Nabila); A.S. Dimas (Antigone); E. Wheeler (Eleanor); H. Chen (Han); B.F. Voight (Benjamin); J. Taneera (Jalal); S. Kanoni (Stavroula); J. Peden (John); F. Turrini (Fabiola); S. Gustafsson (Stefan); C. Zabena (Carina); P. Almgren (Peter); G.V. Dedoussis (George); D. Barnes (Daniel); E.M. Dennison (Elaine); K. Hagen (Knut); P. Eriksson (Per); E. Eury (Elodie); L. Folkersen (Lasse); C.S. Fox (Caroline); T.M. Frayling (Timothy); A. Goel (Anuj); M. Horikoshi (Momoko); B. Isomaa (Bo); A.U. Jackson (Anne); K. Jameson (Karen); E. Kajantie (Eero); J. Kerr-Conte (Julie); L. Groop (Leif); J. Kuusisto (Johanna); R.J.F. Loos (Ruth); J. Luan; K. Makrilakis (Konstantinos); A.K. Manning (Alisa); M.T. Martinez-Larrad (Maria Teresa); N. Narisu (Narisu); J. Öhrvik (John); C. Osmond (Clive); L. Pascoe (Laura); F. Payne (Felicity); A.A. Sayer; B. Sennblad (Bengt); C. Cooper (Charles); K. Stirrups (Kathy); A.J. Swift (Amy); A.C. Syvänen; T. Tuomi (Tiinamaija); F. van't Hooft (Ferdinand); M. Walker (Mark); M.N. Weedon (Michael); W. Xie (Weijia); B. Zethelius (Björn); L.J. Scott (Laura); V. Steinthorsdottir (Valgerdur); A.P. Morris (Andrew); C. Dina (Christian); R.P. Welch (Ryan); E. Zeggini (Eleftheria); C. Huth (Cornelia); Y.S. Aulchenko (Yurii); G. Thorleifsson (Gudmar); L.J. McCulloch (Laura); T. Ferreira (Teresa); H. Grallert (Harald); N. Amin (Najaf); G. Wu (Guanming); C.J. Willer (Cristen); S. Raychaudhuri (Soumya); S.A. McCarroll (Steven); O.M. Hofmann (Oliver); L. Qi (Lu); A.V. Segrè (Ayellet); M. van Hoek (Mandy); P. Navarro (Pau); K.G. Ardlie (Kristin); B. Balkau (Beverley); N. Narisu (Narisu); A.J. Bennett (Amanda); R. Blagieva (Roza); E.A. Boerwinkle (Eric); L.L. Bonnycastle (Lori); K.B. Boström (Kristina Bengtsson); B. Bravenboer (Bert); S. Bumpstead (Suzannah); N.P. Burtt (Noël); G. Charpentier (Guillaume); P.S. Chines (Peter); M. Cornelis (Marilyn); D.J. Couper (David); G. Crawford (Gabe); A.S.F. Doney (Alex); K.S. Elliott (Katherine); A.L. Elliott (Amanda); M.R. Erdos (Michael); C.S. Franklin (Christopher); M. Ganser (Martha); C. Gieger (Christian); N. Grarup (Niels); T. Green (Todd); S. Griffin (Simon); C.J. Groves (Christopher); C. Guiducci (Candace); S. Hadjadj (Samy); N. Hassanali (Neelam); C. Herder (Christian); T. Jorgensen (Torben); W.H.L. Kao (Wen); N. Klopp (Norman); A. Kong (Augustine); P. Kraft (Peter); T. Lauritzen (Torsten); M. Li (Man); A. Lieverse (Aloysius); M.N. Weedon (Michael); V. Lyssenko (Valeriya); M. Marre (Michel); T. Meitinger (Thomas); K. Midthjell (Kristian); M.A. Morken (Mario); P. Nilsson (Peter); K.R. Owen (Katharine); J.R.B. Perry (John); A.K. Petersen; C. Platou (Carl); C. Proença (Christine); W. Rathmann (Wolfgang); R.R. Frants (Rune); G. Rocheleau (Ghislain); M. Roden (Michael); M.J. Sampson (Michael); R. Saxena (Richa); B.M. Shields (Beverley); P. Shrader (Peter); T. Sparsø (Thomas); K. Strassburger (Klaus); H.M. Stringham (Heather); Q. Sun (Qi); B. Thorand (Barbara); J. Tichet (Jean); T.W. van Haeften (Timon); T.W. van Herpt (Thijs); J.V. van Vliet-Ostaptchouk (Jana); G.B. Walters (Bragi); C. Wijmenga (Cisca); S. Cauchi (Stephane); A.L. Gloyn (Anna); U. Gyllensten (Ulf); T. Hansen (T.); W.A. Hide (Winston); G.A. Hitman (Graham); A. Hofman (Albert); K. Hveem (Kristian); M. Laakso (Markku); K.L. Mohlke (Karen L.); A.D. Morris (Andrew); C.N.A. Palmer (Colin); L.D. Stein (Lincoln); J. Tuomilehto (Jaakko); A.G. Uitterlinden (André); R.M. Watanabe (Richard); G.R. Abecasis (Gonçalo); B.O. Boehm (Bernhard); H. Campbell (Harry); M.J. Daly (Mark); A.T. Hattersley (Andrew); F.B. Hu (Frank B.); J.B. Meigs (James); J.S. Pankow (James); O. Pedersen (Oluf); I. Barroso (Inês); L. Groop (Leif); R. Sladek (Rob); U. Thorsteinsdottir (Unnur); J.F. Wilson (James F.); T. Illig (Thomas); P. Froguel (Philippe); C.M. van Duijn (Cock); J-A. Zwart (John-Anker); D. Altshuler (David); M. Boehnke (Michael); M.I. McCarthy (Mark I.); E.K. Speliotes (Elizabeth); S.I. Berndt (Sonja); K.L. Monda (Keri); H.L. Allen; R. Mägi (Reedik); J.C. Randall (Joshua); S. Vedantam (Sailaja); T.W. Winkler (Thomas W.); T. Workalemahu (Tsegaselassie); I.M. Heid (Iris); A.R. Wood (Andrew); R.J. Weyant (Robert); K. Estrada Gil (Karol); L. Liang (Liming); J. Nemesh (James); J.H. Park; T.O. Kilpeläinen (Tuomas); J. Yang (Jian); M.F. Feitosa (Mary Furlan); Z. Kutalik (Zoltán); I. Prokopenko (Inga); W. Rathmann (Wolfgang); A.V. Smith; J.H. Zhao; K.K.H. Aben (Katja); D. Absher (Devin); A.L. Dixon (Anna); B.M. Shields (Beverley); N.L. Glazer (Nicole); N.L. Heard-Costa (Nancy); V. Hoesel (Volker); J.J. Hottenga (Jouke Jan); B. Thorand (Barbara); C. Lamina (Claudia); S. Li (Shengxu); R.M. van Dam (Rob); R.H. Myers (Richard); M.J. Peters (Marjolein); M. Preuss (Michael); S. Ripatti (Samuli); F. Rivadeneira Ramirez (Fernando); C. Sandholt (Camilla); N. Timpson (Nicholas); J.P. Tyrer (Jonathan); S. van Wingerden (Sophie); C.C. White (Charles); F. Wiklund (Fredrik); D.I. Chasman (Daniel); R.W. Lawrence (Robert); N. Pellikka (Niina); J. Shi (Jianxin); E. Thiering (Elisabeth); H. Alavere (Helene); M.T.S. Alibrandi (Maria); A.M. Arnold (Alice); T. Aspelund (Thor); I. Rudan (Igor); E.J.G. Sijbrands (Eric); S.M. Bergmann (Sven); H. Biebermann (Heike); A.I.F. Blakemore (Alexandra); T. Boes (Tanja); S.R. Bornstein (Stefan R.); G.R. Abecasis (Gonçalo); B.O. Boehm (Bernhard); F. Busonero; C. Cavalcanti-Proença (Christine); F.B. Hu (Frank); C.-M. Chen (Chih-Mei); R. Clarke (Robert); J. Connell (John); I.N.M. Day (Ian N.M.); J. Duan (Jubao); R. Elosua (Roberto); G. Eiriksdottir (Gudny); T. Illig (Thomas); S.B. Felix (Stephan); P. Fischer-Posovszky (Pamela); A.R. Folsom (Aaron); N. Friedrich (Nele); M. Fu (Mao); S. Gaget (Stefan); P.V. Gejman (Pablo); E.J. Geus (Eeco); A.P. Gjesing (Anette); P. Goyette (Philippe); J. Gräsler (Jürgen); A.S. Havulinna (Aki); C. Hayward (Caroline); A.C. Heath (Andrew C.); C. Hengstenberg (Christian); A.A. Hicks (Andrew); A. Hinney (Anke); G. Homuth (Georg); J. Hui (Jennie); W. Igl (Wilmar); K.B. Jacobs (Kevin); I. Jarick (Ivonne); E. Jewell (Eelizabeth); U. John (Ulrich); P. Jousilahti (Pekka); A. Jula (Antti); M. Kaakinen (Marika); L. Kaplan (Lee); S. Kathiresan (Sekar); J. Kettunen (Johannes); J. Yang (Joanna); J.W. Knowles (Joshua); T. Esko (Tõnu); I.R. König (Inke); S. Koskinen (Seppo); P. Kovacs (Peter); S. Raychaudhuri (Soumya); J. Laitinen (Jaana); O. Lantieri (Olivier); C. Lanzani (Chiara); L.J. Launer (Lenore); C. Lecoeur (Cécile); T. Lehtimäki (Terho); G. Lettre (Guillaume); J. Liu (Jianjun); M.L. Lokki; M. Lorentzon (Mattias); M.E. Goddard (Michael); B. Ludwig (Barbara); P. Manunta (Paolo); D. Marek (Diana); N.G. Martin (Nicholas); T. Johnson (Toby); B. McKnight (Barbara); O. Melander (Olle); D. Meyre (David); G.W. Montgomery (Grant); R. Mulic (Rosanda); J.S. Ngwa; M. Nelis (Mari); M.J. Neville (Matthew); D.R. Nyholt (Dale); C.J. O'Donnell (Christopher); L.J. Scott (Laura); B.A. Oostra (Ben); G. Pare (Guillame); A.N. Parker (Alex); I. Pichler (Irene); K.H. Pietilainen (Kirsi Hannele); C.P. Platou (Carl); O. Polasek (Ozren); M.N. Cooper (Matthew); S. Rafelt (Suzanne); O.T. Raitakari (Olli T.); N.W. Rayner (Nigel William); M. Ridderstråle (Martin); J. Shi (Jianxin); E. Thiering (Eelisabeth); V. Salomaa (Veikko); M.S. Sandhu (Manjinder); S. Sanna (Serena); J. Saramies (Jouko); M.J. Savolainen (Markku); A. Scherag (Andre); S. Schipf (Sabine); Y. Ben-Shlomo; H. Schunkert (Heribert); K. Silander (Kaisa); J. Sinisalo (Juha); D.S. Siscovick (David); J.H. Smit (Jan); N. Soranzo (Nicole); S.R. Bornstein (Stefan); J. Stephens (Jonathan); T.A. Buchanan (Thomas); M.L. Tammesoo; J.-C. Tardif (Jean-Claude); F.P. Cappuccio (Francesco); T.M. Teslovich (Tanya M.); J.R. Thompson (John); B. Thomson (Brian); A. Tönjes (Anke); R. Clarke; L. Coin (Lachlan); V. Vatin (Vincent); I.N.M. Day (Ian); M. den Heijer (Martin); S. Ebrahim (Shanil); L. Waite (Lindsay); H. Wallaschofski (Henri); E. Widen (Elisabeth); S. Wiegand (Susanna); S.H. Wild (Sarah); G.A.H.M. Willemsen (Gonneke); J.C.M. Witteman (Jacqueline); J. Xu (Jianfeng); L. Zgaga (Lina); J.P. Beilby (John); I.S. Farooqi (I. Sadaf); J. Hebebrand (Johannes); H.V. Huikuri (Heikki); A. James (Alan); M. Kähönen (Mika); F. Macciardi (Fabio); M.S. Nieminen (Markku); C. Ohlsson (Claes); V. Gudnason (Vilmundur); P.M. Ridker (Paul); M. Stumvoll (Michael); J.S. Beckmann (Jacques); D.I. Boomsma (Dorret); M. Caulfield (Mark); S.J. Chanock (Stephen); L.A. Cupples (Adrienne); G.D. Smith; J. Erdmann (Jeanette); H. Grönberg (Henrik); P. Hall (Per); T.B. Harris (Tamara); R.B. Hayes (Richard); J. Heinrich (Joachim); M.-R. Jarvelin (Marjo-Riitta); J. Kaprio (Jaakko); K.T. Khaw; L.A.L.M. Kiemeney (Bart); H. Krude; D.A. Lawlor (Debbie); A. Metspalu (Andres); W.H. Ouwehand (Willem); B.W.J.H. Penninx; A. Peters (Annette); T. Quertermous (Thomas); T. Reinehr (Thomas); A. Rissanen (Aila); N.J. Samani (Nilesh); P.E.H. Schwarz (Peter); A.R. Shuldiner (Alan); T.D. Spector (Timothy); M. Uda (Manuela); Wabitsch, M. (Martin); G. Waeber (Gérard); A.F. Wright (Alan); M.C. Zillikens (Carola); N. Chatterjee (Nilanjan); T. Lehtimäki (Terho); J. Liu (Jianjun); T.L. Assimes (Themistocles); I.B. Borecki (Ingrid); P. Deloukas (Panagiotis); L. Groop (Leif); T. Haritunians (Talin); R.C. Kaplan (Robert); L. Peltonen (Leena Johanna); D.P. Strachan (David); H.E. Wichmann (Heinz Erich); K.E. North (Kari); J.N. Hirschhorn (Joel); E. Ingelsson (Erik); G.W. Montgomery (Grant); L. Parts (Leopold); D. Glass (Daniel); J. Nisbet (James); A. Barrett (Angela); M. Sekowska (Magdalena); M.E. Travers (Mary); S.C. Potter (Simon); E. Grundberg (Elin); S. O'Rahilly (Stephen); A.K. Hedman (Asa); V. Bataille (Veronique); J.T. Bell (Jordana); G. Surdulescu (Gabriela); M. Perola (Markus); F.O. Nestle (Frank); J. Min (Josine); A. Wilk (Alicja); C.J. Hammond (Christopher J.); T.-P. Yang (Tsun-Po); O. Raitakari (Olli); R. Durbin (Richard); K.R. Ahmadi (Kourosh); H. Holm (Hilma); A.F. Stewart (Alexandre F.); M. Barbalic (maja); Z. Aherrahrou (Zouhair); H. Allayee (Hooman); S.S. Anand (Sonia); K. Andersen (Karl); S. Schreiber (Stefan); D. Ardissino (Diego); T.A. Barnes (Timothy); D.M. Becker (Diane); L.C. Becker (Lewis); K. Berger (Klaus); J.C. Bis (Joshua); S.M. Boekholdt (Matthijs); P.S. Braund (Peter); M.S. Burnett; I. Buysschaert (Ian); J.F. Carlquist (John); L. Chen (Li); S. Cichon (Sven); V. Codd (Veryan); R.W. Davies (Robert); G.V. Dedoussis (George); A. Dehghan (Abbas); S. Demissie (Serkalem); P. Diemert (Patrick); R. Do (Ron); A. Doering (Angela); S. Eifert (Sandra); N.E. El Mokhtari (Nour Eddine); S.G. Ellis (Stephen); S.E. Epstein (Stephen); U. de Faire (Ulf); M. Fischer (Marcus); J. Freyer (Jennifer); B. Gigante (Bruna); D. Girelli (Domenico); D.R. Witte (Deniel); J.R. Gulcher (Jeffrey); E. Halperin (Eran); N. Hammond (Naomi); S.L. Hazen (Stanley); A. Ziegler (Andreas); G.T. Jones (Gregory); J.W. Jukema (Jan Wouter); I.S. Farooqi (Sadaf); J.J.P. Kastelein (John); R. Laaksonen (Reijo); D. Lambrechts (Diether); D.F. Levinson (Douglas); X. Li (Xiaohui); W. Lieb (Wolfgang); C. Loley (Christina); A.J. Lotery (Andrew); P.M. Mannucci (Pier); S. Maouche (Seraya); J.S. Beckmann (Jacques); H. Boeing (Heiner); C. Meisinger (Christa); V. Mooser (Vincent); T. Morgan (Thomas); F.S. Collins (Francis); J.B. Muhlestein (Joseph); T. Munzel (Thomas); K. Musunuru (Kiran); J. Nahrstaedt (Janja); C.P. Nelson (Christopher P.); M.M. Nöthen (Markus); R.S. Patel (Riyaz); F. Peyvandi (Flora); R.B. Hayes (Richard); A.A. Quyyumi (Arshed); D.J. Rader (Daniel); L.S. Rallidis (Loukianos); F. Karpe (Fredrik); J. Kaprio (Jaakko); M.L. Sampietro (Maria Lourdes); M.S. Sandhu (Manjinder); E.E. Schadt (Eric); A. Schäfer (Arne); A. Schillert (Arne); S.M. Schwartz (Stephen); P. Munroe (Patricia); S. Sivapalaratnam (Suthesh); A.V. Smith (Albert Vernon); J.D. Snoep (Jaapjan); J.A. Spertus (John); K. Stark (Klaus); M. Stoll (Monika); W. Tang (W.); S. Tennstedt (Stephanie); G. Thorgeirsson (Gudmundur); A.R. Shuldiner (Alan); A.M. van Rij (Andre); N.J. Wareham (Nick); G.A. Wells (George); P.S. Wild (Philipp); C. Willenborg (Christina); B.J. Wright (Benjamin); T. Zeller (Tanja); F. Cambien (François); A.H. Goodall (Alison); W. März (Winfried); S. Blankenberg (Stefan); R. Roberts (Robert); R. McPherson (Ruth); J. Hopewell; P.M. Visscher (Peter); A. Offer (Alison); L. Bowman; P. Sleight (Peter); R. Peto (R.); F.S. Collins (Francis); J.C. Chambers (John C.); N. Ahmed (Nabeel); J.R. O´Connell; P. Donnelly (Peter); J.S. Kooner (Jaspal); N.J. Samani (Nilesh); J. Scott (James); J.S. Sehmi (Joban); W. Zhang (Weihua); R.J. Strawbridge (Rona); Sabater-Lleal, M. (Maria); A. Mälarstig (Anders); M.-L. Hellénius (Mai-Lis); G. Olsson; S. Rust (Stephan); G. Assmann (Gerd); U. Seedorf (Udo); G. Tognoni; M. Franzosi; P. Linksted (Pamela); H. Ongen (Halit); T. Kyriakou (Theodosios); M. Farrall (Martin); A. Rasheed (Asif); M.A. Zaidi (Aghar); N. Shah (Nisha); M. Samuel (Maria); C.B. Mallick (Chandana Basu); M. Azhar (Muhammad); K.S. Zaman (Khan Shah); M. Ishaq (Muhammad); A. Gardezi (Ali); C.J. Hammond (Christopher); R. Frossard; J. Danesh (John); J.C. Chambers (John); J.S. Kooner (Jaspal S.); C.-G. Östenson (Claes-Göran); K.T. Zondervan (Krina); M. Serrano-Ríos (Manuel); E. Ferrannini (Ele); T. Forsen (Tom); M.I. McCarthy (Mark); G.V. Dedoussis (George); C. Langenberg (Claudia); A. Hamsten (Anders); J.C. Florez (Jose)

    2011-01-01

    textabstractOBJECTIVE - Proinsulin is a precursor of mature insulin and C-peptide. Higher circulating proinsulin levels are associated with impaired b-cell function, raised glucose levels, insulin resistance, and type 2 diabetes (T2D). Studies of the insulin processing pathway could provide new

  8. Genome-Wide Association Identifies Nine Common Variants Associated With Fasting Proinsulin Levels and Provides New Insights Into the Pathophysiology of Type 2 Diabetes

    NARCIS (Netherlands)

    Strawbridge, Rona J.; Dupuis, Josee; Prokopenko, Inga; Barker, Adam; Ahlqvist, Emma; Rybin, Denis; Petrie, John R.; Travers, Mary E.; Bouatia-Naji, Nabila; Dimas, Antigone S.; Nica, Alexandra; Wheeler, Eleanor; Chen, Han; Voight, Benjamin F.; Taneera, Jalal; Kanoni, Stavroula; Peden, John F.; Turrini, Fabiola; Gustafsson, Stefan; Zabena, Carina; Almgren, Peter; Barker, David J. P.; Barnes, Daniel; Dennison, Elaine M.; Eriksson, Johan G.; Eriksson, Per; Eury, Elodie; Folkersen, Lasse; Fox, Caroline S.; Frayling, Timothy M.; Goel, Anuj; Gu, Harvest F.; Horikoshi, Momoko; Isomaa, Bo; Jackson, Anne U.; Jameson, Karen A.; Kajantie, Eero; Kerr-Conte, Julie; Kuulasmaa, Teemu; Kuusisto, Johanna; Loos, Ruth J. F.; Luan, Jian'an; Makrilakis, Konstantinos; Manning, Alisa K.; Teresa Martinez-Larrad, Maria; Narisu, Narisu; Mannila, Maria Nastase; Ohrvik, John; Osmond, Clive; Pascoe, Laura

    2011-01-01

    OBJECTIVE-Proinsulin is a precursor of mature insulin and C-peptide. Higher circulating proinsulin levels are associated with impaired beta-cell function, raised glucose levels, insulin resistance, and type 2 diabetes (T2D). Studies of the insulin processing pathway could provide new insights about

  9. Genome-wide association identifies nine common variants associated with fasting proinsulin levels and provides new insights into the pathophysiology of type 2 diabetes

    NARCIS (Netherlands)

    Strawbridge, R.J.; Dupuis, J.; Prokopenko, I.; Barker, A.; Ahlqvist, E.; Rybin, D.; Petrie, J.R.; Travers, M.E.; Bouatia-Naji, N.; Dimas, A.S.; Nica, A.; Wheeler, E.; Chen, H.; Voight, B.F.; Taneera, J.; Kanoni, S.; Peden, J.F.; Turrini, F.; Gustafsson, S.; Zabena, C.; Almgren, P.; Barker, D.J.; Barnes, D.; Dennison, E.M.; Eriksson, J.G.; Eriksson, P.; Eury, E.; Folkersen, L.; Fox, C.S.; Frayling, T.M.; Goel, A.; Gu, H.F.; Horikoshi, M.; Isomaa, B.; Jackson, A.U.; Jameson, K.A.; Kajantie, E.; Kerr-Conte, J.; Kuulasmaa, T.; Kuusisto, J.; Loos, R.J.; Luan, J.; Makrilakis, K.; Manning, A.K.; Martinez-Larrad, M.T.; Narisu, N.; Nastase Mannila, M.; Ohrvik, J.; Osmond, C.; Pascoe, L.; Payne, F.; Sayer, A.A.; Sennblad, B.; Silveira, A.; Stancakova, A.; Stirrups, K.; Swift, A.J.; Syvanen, A.C.; Tuomi, T.; Hooft, F. van 't; Walker, M.; Weedon, M.N.; Xie, W.; Zethelius, B.; Ongen, H.; Malarstig, A.; Hopewell, J.C.; Saleheen, D.; Chambers, J.; Parish, S.; Danesh, J.; Kooner, J.; Ostenson, C.G.; Lind, L.; Cooper, C.C.; Serrano-Rios, M.; Ferrannini, E.; Forsen, T.J.; Clarke, R.; Franzosi, M.G.; Seedorf, U.; Watkins, H.; Froguel, P.; Johnson, P.; Deloukas, P.; Collins, F.S.; Laakso, M.; Dermitzakis, E.T.; Boehnke, M.; McCarthy, M.I.; Wareham, N.J.; Groop, L.; Pattou, F.; Gloyn, A.L.; Dedoussis, G.V.; Lyssenko, V.; Meigs, J.B.; Barroso, I.; Watanabe, R.M.; Heijer, M. den; Kiemeney, L.A.L.M.; et al.,

    2011-01-01

    OBJECTIVE: Proinsulin is a precursor of mature insulin and C-peptide. Higher circulating proinsulin levels are associated with impaired beta-cell function, raised glucose levels, insulin resistance, and type 2 diabetes (T2D). Studies of the insulin processing pathway could provide new insights about

  10. Genome-wide association identifies nine common variants associated with fasting proinsulin levels and provides new insights into the pathophysiology of type 2 diabetes

    NARCIS (Netherlands)

    Strawbridge, Rona J.; Dupuis, Josée; Prokopenko, Inga; Barker, Adam; Ahlqvist, Emma; Rybin, Denis; Petrie, John R.; Travers, Mary E.; Bouatia-Naji, Nabila; Dimas, Antigone S.; Nica, Alexandra; Wheeler, Eleanor; Chen, Han; Voight, Benjamin F.; Taneera, Jalal; Kanoni, Stavroula; Peden, John F.; Turrini, Fabiola; Gustafsson, Stefan; Zabena, Carina; Almgren, Peter; Barker, David J. P.; Barnes, Daniel; Dennison, Elaine M.; Eriksson, Johan G.; Eriksson, Per; Eury, Elodie; Folkersen, Lasse; Fox, Caroline S.; Frayling, Timothy M.; Goel, Anuj; Gu, Harvest F.; Horikoshi, Momoko; Isomaa, Bo; Jackson, Anne U.; Jameson, Karen A.; Kajantie, Eero; Kerr-Conte, Julie; Kuulasmaa, Teemu; Kuusisto, Johanna; Loos, Ruth J. F.; Luan, Jian'an; Makrilakis, Konstantinos; Manning, Alisa K.; Martínez-Larrad, María Teresa; Narisu, Narisu; Nastase Mannila, Maria; Boekholdt, S. Matthijs; Kastelein, John J. P.; Rosendaal, Frits R.

    2011-01-01

    Proinsulin is a precursor of mature insulin and C-peptide. Higher circulating proinsulin levels are associated with impaired β-cell function, raised glucose levels, insulin resistance, and type 2 diabetes (T2D). Studies of the insulin processing pathway could provide new insights about T2D

  11. Clinical significance of changes of serum true insulin and proinsulin levels in relations of patients with type 2 diabetes mellitus

    International Nuclear Information System (INIS)

    Tian Xiaoping; Huang Huijian; Huang Haibo; Wu Yan; He Haoming

    2004-01-01

    Objective: To explore the degree of insulin resistance and β-cell secretory function impairment in close (1st degree) relations of patients with type 2 diabetes (DMII). Methods: Serum true insulin (TI), pro-insulin (PI), immunoreactive insulin (IRI) levels at fasting and after oral 75g glucose loading were determined in: 1) patients with DM 2, n=65 2)relations of DM 2 patients with impaired glucose tolerance (IGT), n=34 3) relations of DM 2 patients with normal glucose tolerance (NGT), n=66 and 4) controls, n=48. HOMA-IR and HOMA-β cell secretory indices were calculated from the data. Results: Fasting serum PI levels were significantly higher in DM 2 patients, relations with IGT and NGT than those in the controls (t=2.38, t=2.16, t=1.95, P 1 C percentages were significantly higher in DM 2 patients and IGT, NGT groups than those in controls (t=3.67, t=2.45, t=1.97, P 1 C percentage, fasting TI and IRI levels. Conclusion: Insulin resistance was already obvious in those relations of DM 2 patients with normal glucose tolerance and β-cell secretory function impairment was also present. Early intervention in these subjects might be beneficial. (authors)

  12. Proinsulin atypical maturation and disposal induces extensive defects in mouse Ins2+/Akita β-cells.

    Directory of Open Access Journals (Sweden)

    Qingxin Yuan

    Full Text Available Because of its low relative folding rate and plentiful manufacture in β-cells, proinsulin maintains a homeostatic balance of natively and plentiful non-natively folded states (i.e., proinsulin homeostasis, PIHO through the integration of maturation and disposal processes. PIHO is susceptible to genetic and environmental influences, and its disorder has been critically linked to defects in β-cells in diabetes. To explore this hypothesis, we performed polymerase chain reaction (PCR, metabolic-labeling, immunoblotting, and histological studies to clarify what defects result from primary disorder of PIHO in model Ins2(+/Akita β-cells. We used T antigen-transformed Ins2(+/Akita and control Ins2(+/+ β-cells established from Akita and wild-type littermate mice. In Ins2(+/Akita β-cells, we found no apparent defect at the transcriptional and translational levels to contribute to reduced cellular content of insulin and its precursor and secreted insulin. Glucose response remained normal in proinsulin biosynthesis but was impaired for insulin secretion. The size and number of mature insulin granules were reduced, but the size/number of endoplasmic reticulum, Golgi, mitochondrion, and lysosome organelles and vacuoles were expanded/increased. Moreover, cell death increased, and severe oxidative stress, which manifested as increased reactive oxygen species, thioredoxin-interacting protein, and protein tyrosine nitration, occurred in Ins2(+/Akita β-cells and/or islets. These data show the first clear evidence that primary PIHO imbalance induces severe oxidative stress and impairs glucose-stimulated insulin release and β-cell survival as well as producing other toxic consequences. The defects disclosed/clarified in model Ins2(+/Akita β-cells further support a role of the genetic and stress-susceptible PIHO disorder in β-cell failure and diabetes.

  13. Proinsulin, GLP-1, and glucagon are associated with partial remission in children and adolescents with newly diagnosed type 1 diabetes

    DEFF Research Database (Denmark)

    Kaas, A.; Andersen, M. L. M.; Fredheim, Siri

    2012-01-01

    .002) were significantly lower in remitters than in non-remitters at 6 and 12 months. Proinsulin associated positively with GLP-1 at 1 month (p = 0.004) and negatively at 6 (p = 0.002) and 12 months (p = 0.0002). Conclusions: In type 1 diabetes, patients in partial remission have higher levels of proinsulin......1C), glucagon-like peptide-1 (GLP-1), glucagon, and remission status the first year after diagnosis of type 1 diabetes. Methods: Juvenile patients (n = 275) were followed 1, 6, and 12 months after diagnosis. At each visit, partial remission was defined as IDAA1C = 9%. The patients had a liquid meal...

  14. Sulfatide promotes the folding of proinsulin, preserves insulin crystals, and mediates its monomerization.

    Science.gov (United States)

    Osterbye, T; Jørgensen, K H; Fredman, P; Tranum-Jensen, J; Kaas, A; Brange, J; Whittingham, J L; Buschard, K

    2001-06-01

    Sulfatide is a glycolipid that has been associated with insulin-dependent diabetes mellitus. It is present in the islets of Langerhans and follows the same intracellular route as insulin. However, the role of sulfatide in the beta cell has been unclear. Here we present evidence suggesting that sulfatide promotes the folding of reduced proinsulin, indicating that sulfatide possesses molecular chaperone activity. Sulfatide associates with insulin by binding to the insulin domain A8--A10 and most likely by interacting with the hydrophobic side chains of the dimer-forming part of the insulin B-chain. Sulfatide has a dual effect on insulin. It substantially reduces deterioration of insulin hexamer crystals at pH 5.5, conferring stability comparable to those in beta cell granules. Sulfatide also mediates the conversion of insulin hexamers to the biological active monomers at neutral pH, the pH at the beta-cell surface. Finally, we report that inhibition of sulfatide synthesis with chloroquine and fumonisine B1 leads to inhibition of insulin granule formation in vivo. Our observations suggest that sulfatide plays a key role in the folding of proinsulin, in the maintenance of insulin structure, and in the monomerization process.

  15. Proinsulin in Healthy Pregnancy, Pregnancy with Gestational Diabetes and after Delivery

    Directory of Open Access Journals (Sweden)

    Genova M. P.

    2014-06-01

    Full Text Available The aim of the present study was to evaluate the levels of pro-insulin and pro-insulin/ insulin ratio (PIR in pregnant with normal glucose tolerance (NGT, pregnant with gestational diabetes mellitus (GDM and women after delivery with GDM history. Normal pregnancy is characterized by progressive insulin resistance, which is physiologically compensated by an increase in insulin secretion. The higher secretion of the insulin precursor pro-insulin has been associated with β-cell dysfunction. A total of 102 pregnant women between 24-28 gestational weeks (53 GDM pregnant, 49 with NGT and 22 postpartum with GDM history, as assessed by a 2h oral glucose tolerance test, were included in the study. Fasting plasma insulin and pro-insulin (PI concentrations at the basal state were measured in all women. The ratio pro-insulin/insulin was calculated. BMI was significantly higher in GDM pregnant compared to NGT weight-matched group (30.56 ± 6.9 vs. 30.56 ± 6.9; p < 0, 011 and compared to the levels after delivery (30.56 ± 6.9vs. 27.9 ± 6, 27; p < 0, 001. Significant differences in the levels of PI between NGT and GDM pregnant (3.94 ± 2.78 vs. 7.59 ± 5.27; p = 0.006, between GDM and postpartum women (7.59 ± 5.27 vs. 4.46 ± 1.14; p = 0.022 were established. No signifi cant difference in the level of PIR between two pregnant groups was observed. Separately NGT and GDM showed signifi cant difference compared to young mothers (0.41 ± 0.14 vs. 0.148 ± 0.031, p < 0.02; 0.46 ± 0.16 vs. 0.148 ± 0.031, p = 0.009. Fasting insulin was statistically higher in GDM pregnant compare to NGT and women after delivery (13.84 ± 8.43 vs. 11.35 ± 7.38, p = 0.02; 13.84 ± 8.43 vs. 10.60 ± 7.53, p < 0.01. The correlation between PIR and BMI in the three groups studied were r = 0.416; r = 0,741; r = 0,556 (with statistical significance p = 0.01 between NGT and GDM pregnancy, p = 0.02 between GDM pregnancy and postpartum, p < 0.0001 between NGT pregnancy and young mother

  16. Trace elements determination in human hair

    International Nuclear Information System (INIS)

    Carrion, Jose

    1995-01-01

    Concentrations of Cu, Zn, Pb, Mg, Ca, Na, K, Mn, Cr, Ni, Co, V, Cd and Al, in human hair sampled from 23 young men during 24 months were determined by atomic absorption spectroscopy. Additional determination of mercury and volatile elements were made by using accessory MHS-10. Statistical treatment of data is presented for each person and element. The pre-treatment of hair carried out with an organic solvent to remove the superficial pollutants is explained. (The author)

  17. Genome-wide association identifies nine common variants associated with fasting proinsulin levels and provides new insights into the pathophysiology of type 2 diabetes

    OpenAIRE

    Strawbridge, Rona; Dupuis, Josée; Prokopenko, Inga; Barker, Adam; Ahlqvist, Emma; Rybin, Denis; Petrie, John; Bouatia-Naji, Nabila; Dimas, Antigone; Wheeler, Eleanor; Chen, Han; Voight, Benjamin; Taneera, Jalal; Kanoni, Stavroula; Peden, John

    2011-01-01

    textabstractOBJECTIVE - Proinsulin is a precursor of mature insulin and C-peptide. Higher circulating proinsulin levels are associated with impaired b-cell function, raised glucose levels, insulin resistance, and type 2 diabetes (T2D). Studies of the insulin processing pathway could provide new insights about T2D pathophysiology. RESEARCH DESIGN AND METHODS - We have conducted a meta-analysis of genome-wide association tests of ;2.5 million genotyped or imputed single nucleotide polymorphisms...

  18. Intact proinsulin and beta-cell function in lean and obese subjects with and without type 2 diabetes

    DEFF Research Database (Denmark)

    Røder, M E; Dinesen, B; Hartling, S G

    1999-01-01

    , total proinsulin immunoreactivity (PIM), intact insulin, and C-peptide (by radioimmunoassay) by specific enzyme-linked immunosorbent assays in the fasting state and during a 120-min glucagon (1 mg i.v.) stimulation test. Lean (BMI 23.5 +/- 0.3 kg/m2) (LD) and obese (30.1 +/- 0.4 kg/m2) (OD) type 2...... diabetic patients matched for fasting glucose (10.2 +/- 0.6 vs. 10.3 +/- 0.4 mmol/l) were compared with age- and BMI-matched lean (22.4 +/- 0.6 kg/m2) (LC) and obese (30.8 +/- 0.9 kg/m2) (OC) normal control subjects. RESULTS: Diabetic patients (LD vs. LC and OD vs. OC) had elevated fasting levels of intact......, most pronounced in the lean group. The ratio of intact proinsulin to PIM was higher in diabetic patients after stimulation in both LD versus LC: 32 +/- 3 vs. 23 +/- 2%, and OD versus OC: 28 +/- 4 vs. 16 +/- 2%, both P obese normal subjects, intact proinsulin/PIM was lower both in the fasting...

  19. Optical spectroscopic determination of human meniscus composition.

    Science.gov (United States)

    Ala-Myllymäki, Juho; Honkanen, Juuso T J; Töyräs, Juha; Afara, Isaac O

    2016-02-01

    This study investigates the correlation between the composition of human meniscus and its absorption spectrum in the visible (VIS) and near infrared (NIR) spectral range. Meniscus samples (n = 24) were obtained from nonarthritic knees of human cadavers with no history of joint diseases. Specimens (n = 72) were obtained from three distinct sections of the meniscus, namely; anterior, center, posterior. Absorption spectra were acquired from each specimen in the VIS and NIR spectral range (400-1,100 nm). Following spectroscopic probing, the specimens were subjected to biochemical analyses to determine the matrix composition, that is water, hydroxyproline, and uronic acid contents. Multivariate analytical techniques, including principal component analysis (PCA) and partial least squares (PLS) regression, were then used to investigate the correlation between the matrix composition and it spectral response. Our results indicate that the optical absorption of meniscus matrix is related to its composition, and this relationship is optimal in the NIR spectral range (750-1,100 nm). High correlations (R(2) (uronic)  = 86.9%, R(2) (water)  = 83.8%, R(2) (hydroxyproline)  = 81.7%, p meniscus composition, thus suggesting that spectral data in the NIR range can be utilized for estimating the matrix composition of human meniscus. In conclusion, optical spectroscopy, particularly in the NIR spectral range, is a potential method for evaluating the composition of human meniscus. This presents a promising technique for rapid and nondestructive evaluation of meniscus integrity in real-time during arthroscopic surgery. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  20. Stimulation by ATP of proinsulin to insulin conversion in isolated rat pancreatic islet secretory granules. Association with the ATP-dependent proton pump

    International Nuclear Information System (INIS)

    Rhodes, C.J.; Lucas, C.A.; Mutkoski, R.L.; Orci, L.; Halban, P.A.

    1987-01-01

    Isolated rat pancreatic islets were pulse-labeled for 5 min with [ 3 H]leucine then chased for 25 min, during which time endogenously labeled [ 3 H]proinsulin becomes predominantly compartmented in immature secretory granules. The islets were then homogenized in isotonic sucrose (pH 7.4) and a beta-granule preparation obtained by differential centrifugation and discontinuous sucrose gradient ultracentrifugation. This preparation was enriched 8-fold in beta-granules. Aside from contamination with mitochondria and a limited number of lysosomes, the beta-granule preparation was essentially free of any other organelles involved in proinsulin synthesis and packaging (i.e. microsomal elements and, more particularly, Golgi complex). Conversion of endogenously labeled [ 3 H]proinsulin was followed in this beta-granule fraction for up to 2 h at 37 degrees C in a buffer (pH 7.3) that mimicked the cationic constituents of B-cell cytosol, during which time 92% of the beta-granules remained intact. Proinsulin conversion was analyzed by high performance liquid chromatography. The rate of proinsulin conversion to insulin was stimulated by 2.2 +/- 0.1-fold (n = 6) (at a 60-min incubation) in the presence of ATP (2 mM) and an ATP regenerating system compared to beta-granule preparations incubated without ATP. This ATP stimulation was abolished in the presence of beta-granule proton pump ATPase inhibitors (tributyltin, 2.5 microM, or 1,3-dicyclohexylcarbodiimide, 50 microM). Inhibitors of mitochondrial proton pump ATPases had no effect on the ATP stimulation of proinsulin conversion. When granules were incubated in a more acidic buffer, proinsulin conversion was increased relative to that at pH 7.3. At pH 5.5, ATP no longer stimulated conversion, and tributyltin and 1,3-dicyclohexylcarbodiimide had no effect

  1. DETERMINANTS OF GLOBAL DEVELOPMENT IN HUMAN MOTIVATION

    Directory of Open Access Journals (Sweden)

    ŢÂMPU DIANA LARISA

    2015-06-01

    Full Text Available We live in a world were manner of use of information is crucial in determining the level of performance. Each country around the globe uses a proper way of spreading information and communication. Studies present the Information and Communication Technology (ICT indicator the proper tool to provide an objective evaluation of the countries performance. The question that this research wants to answer is what are the main ways of motivation (extrinsic and intrinsic in countries where ICT Development Index reaches the highest values in the last 2 years. In this way, we want to verify if ICT has different predictors and different possible consequence that depend on human motivation. Thus relying on calculations made by the International Telecommunication Union for ICT and key factors of motivation this paper will present if there is any relationship between citizens motivation and ICT. This hypothesized model will be illustrated with data from thirty developed countries.

  2. Sequence determinants of human microsatellite variability

    Directory of Open Access Journals (Sweden)

    Jakobsson Mattias

    2009-12-01

    Full Text Available Abstract Background Microsatellite loci are frequently used in genomic studies of DNA sequence repeats and in population studies of genetic variability. To investigate the effect of sequence properties of microsatellites on their level of variability we have analyzed genotypes at 627 microsatellite loci in 1,048 worldwide individuals from the HGDP-CEPH cell line panel together with the DNA sequences of these microsatellites in the human RefSeq database. Results Calibrating PCR fragment lengths in individual genotypes by using the RefSeq sequence enabled us to infer repeat number in the HGDP-CEPH dataset and to calculate the mean number of repeats (as opposed to the mean PCR fragment length, under the assumption that differences in PCR fragment length reflect differences in the numbers of repeats in the embedded repeat sequences. We find the mean and maximum numbers of repeats across individuals to be positively correlated with heterozygosity. The size and composition of the repeat unit of a microsatellite are also important factors in predicting heterozygosity, with tetra-nucleotide repeat units high in G/C content leading to higher heterozygosity. Finally, we find that microsatellites containing more separate sets of repeated motifs generally have higher heterozygosity. Conclusions These results suggest that sequence properties of microsatellites have a significant impact in determining the features of human microsatellite variability.

  3. LC determination of praziquantel in human plasma.

    Science.gov (United States)

    Ridtitid, Wibool; Wongnawa, Malinee; Mahatthanatrakul, Werawath; Punyo, Jarurat; Sunbhanich, Methi

    2002-04-01

    A simple high-performance liquid chromatographic (HPLC) method for the determination of praziquantel in human plasma was developed and validated. The present method was described by adding drop-wise 0.2 M Zinc sulfate and acetonitrile to plasma sample for deproteinization. This method used a reversed-phase Spherisorb ODS 2 column (5 microm), 250 x 4.6 mm i.d. as a stationary phase with a mobile phase consisting of acetonitrile- methanol-water (36:10:54, v/v/v), a flow rate of 1.5 ml/min and UV detection wavelength of 217 nm. Diazepam was used as internal standard. The standard calibration curve was linear over the concentration range of 100-2000 ng/ml (r=0.999). The equation of a linear regression line was y=8.05E-04+7.25E-04x with slope and intercept values of 0.0007 and 0.0008, respectively. The limit of detection was 12.25 ng/ml and the limit of quantification was set at 100 ng/ml. The intra- and inter-day assay coefficients of variation (CV) were 3.0+/-1.7 and 6.3+/-1.9%, respectively. The percentage of recovery was 102.1+/-5.6. Therefore, the HPLC method described here was simple, rapid and reproducible since it did not require extraction and evaporation processes in sample preparation, which will reduce time-consuming or expensive sample preparation.

  4. Characterization and Oral Delivery of Proinsulin-Transferrin Fusion Protein Expressed Using ExpressTec

    Directory of Open Access Journals (Sweden)

    Yu-Sheng Chen

    2018-01-01

    Full Text Available Proinsulin-transferrin fusion protein (ProINS-Tf has been designed and successfully expressed from the mammalian HEK293 cells (HEK-ProINS-Tf. It was found that HEK-ProINS-Tf could be converted into an activated form in the liver. Furthermore, HEK-ProINS-Tf was demonstrated as an extra-long acting insulin analogue with liver-specific insulin action in streptozotocin (STZ-induced type 1 diabetic mice. However, due to the low production yield from transfected HEK293 cells, there are other interesting features, including the oral bioavailability, which have not been fully explored and characterized. To improve the protein production yield, an alternative protein expression system, ExpressTec using transgenic rice (Oryza sativa L., was used. The intact and active rice-derived ProINS-Tf (ExpressTec-ProINS-Tf was successfully expressed from the transgenic rice expression system. Our results suggested that, although the insulin-like bioactivity of ExpressTec-ProINS-Tf was slightly lower in vitro, its potency of in vivo blood glucose control was considerably stronger than that of HEK-ProINS-Tf. The oral delivery studies in type 1 diabetic mice demonstrated a prolonged control of blood glucose to near-normal levels after oral administration of ExpressTec-ProINS-Tf. Results in this report suggest that ExpressTec-ProINS-Tf is a promising insulin analog with advantages including low cost, prolonged and liver targeting effects, and most importantly, oral bioactivity.

  5. Species and prevalence determination of Human Intestinal ...

    African Journals Online (AJOL)

    ADOWIE PERE

    harm to some extent by the association or may cause death. Human ... Center and Specialist Hospital Yola, Adamawa state. MATERIALS ..... of animal dungs as manure could aid in transfer of cysts. .... American Journal of Tropical. Medicine ...

  6. The effect of different volumes of high-intensity interval training on proinsulin in participants with the metabolic syndrome: a randomised trial.

    Science.gov (United States)

    Ramos, Joyce S; Dalleck, Lance C; Borrani, Fabio; Mallard, Alistair R; Clark, Bronwyn; Keating, Shelley E; Fassett, Robert G; Coombes, Jeff S

    2016-11-01

    The continuous demand for insulin in the face of insulin resistance, coupled with the glucolipotoxic environment associated with the metabolic syndrome (MetS), adversely affects the quality of insulin produced and secreted by the pancreatic beta cells. This is depicted by increased circulating intact proinsulin concentration, which is associated with increased MetS severity and risk of cardiovascular (CV) mortality. High-intensity interval training (HIIT) has been shown to reduce insulin resistance and other CV disease risk factors to a greater degree than moderate-intensity continuous training (MICT). We therefore aimed to investigate the impact of MICT and different volumes of HIIT on circulating intact proinsulin concentration. This was a substudy of the 'Exercise in prevention of Metabolic Syndrome' (EX-MET) multicentre trial. Sixty-six individuals with MetS were randomised to 16 weeks of: (1) MICT (n = 21, 30 min at 60-70% peak heart rate [HRpeak], five times/week); (2) 4HIIT (n = 22, 4 × 4 min bouts at 85-95% HRpeak, interspersed with 3 min of active recovery at 50-70% HRpeak, three times/week); or (3) 1HIIT (n = 23, 1 × 4 min bout at 85-95% HRpeak, three times/week). A subanalysis investigated the differential impact of these training programmes on intact proinsulin concentration in MetS individuals with type 2 diabetes (MICT, n = 6; 4HIIT, n = 9; 1HIIT, n = 12) and without type 2 diabetes (MICT, n = 15; 4HIIT, n = 13; 1HIIT, n = 11). Intact proinsulin, insulin and C-peptide concentrations were measured in duplicate via ELISA, following a 12 h fast, before and after the exercise programme. Fasting intact proinsulin concentration was also expressed relative to insulin and C-peptide concentrations. Following the exercise training, there were no significant (p > 0.05) changes in fasting intact proinsulin concentration indices in all participants (pre- vs post-programme proinsulin, proinsulin

  7. Physiological determinants of human acute hypoxia tolerance.

    Science.gov (United States)

    2013-11-01

    AbstractIntroduction. We investigated possible physiological determinants of variability in hypoxia tolerance in subjects given a 5-minute normobaric exposure to 25,000 ft equivalent. Physiological tolerance to hypoxia was defined as the magnitude of...

  8. Expression of cholera toxin B-proinsulin fusion protein in lettuce and tobacco chloroplasts--oral administration protects against development of insulitis in non-obese diabetic mice.

    Science.gov (United States)

    Ruhlman, Tracey; Ahangari, Raheleh; Devine, Andrew; Samsam, Mohtahsem; Daniell, Henry

    2007-07-01

    Lettuce and tobacco chloroplast transgenic lines expressing the cholera toxin B subunit-human proinsulin (CTB-Pins) fusion protein were generated. CTB-Pins accumulated up to ~16% of total soluble protein (TSP) in tobacco and up to ~2.5% of TSP in lettuce. Eight milligrams of powdered tobacco leaf material expressing CTB-Pins or, as negative controls, CTB-green fluorescent protein (CTB-GFP) or interferon-GFP (IFN-GFP), or untransformed leaf, were administered orally, each week for 7 weeks, to 5-week-old female non-obese diabetic (NOD) mice. The pancreas of CTB-Pins-treated mice showed decreased infiltration of cells characteristic of lymphocytes (insulitis); insulin-producing beta-cells in the pancreatic islets of CTB-Pins-treated mice were significantly preserved, with lower blood or urine glucose levels, by contrast with the few beta-cells remaining in the pancreatic islets of the negative controls. Increased expression of immunosuppressive cytokines, such as interleukin-4 and interleukin-10 (IL-4 and IL-10), was observed in the pancreas of CTB-Pins-treated NOD mice. Serum levels of immunoglobulin G1 (IgG1), but not IgG2a, were elevated in CTB-Pins-treated mice. Taken together, T-helper 2 (Th2) lymphocyte-mediated oral tolerance is a likely mechanism for the prevention of pancreatic insulitis and the preservation of insulin-producing beta-cells. This is the first report of expression of a therapeutic protein in transgenic chloroplasts of an edible crop. Transplastomic lettuce plants expressing CTB-Pins grew normally and transgenes were maternally inherited in T(1) progeny. This opens up the possibility for the low-cost production and delivery of human therapeutic proteins, and a strategy for the treatment of various other autoimmune diseases.

  9. Expression of cholera toxin B–proinsulin fusion protein in lettuce and tobacco chloroplasts – oral administration protects against development of insulitis in non-obese diabetic mice

    Science.gov (United States)

    Ruhlman, Tracey; Ahangari, Raheleh; Devine, Andrew; Samsam, Mohtahsem; Daniell, Henry

    2008-01-01

    Summary Lettuce and tobacco chloroplast transgenic lines expressing the cholera toxin B subunit–human proinsulin (CTB-Pins) fusion protein were generated. CTB-Pins accumulated up to ~16% of total soluble protein (TSP) in tobacco and up to ~2.5% of TSP in lettuce. Eight milligrams of powdered tobacco leaf material expressing CTB-Pins or, as negative controls, CTB–green fluorescent protein (CTB-GFP) or interferon–GFP (IFN-GFP), or untransformed leaf, were administered orally, each week for 7 weeks, to 5-week-old female non-obese diabetic (NOD) mice. The pancreas of CTB-Pins-treated mice showed decreased infiltration of cells characteristic of lymphocytes (insulitis); insulin-producing β-cells in the pancreatic islets of CTB-Pins-treated mice were significantly preserved, with lower blood or urine glucose levels, by contrast with the few β-cells remaining in the pancreatic islets of the negative controls. Increased expression of immunosuppressive cytokines, such as interleukin-4 and interleukin-10 (IL-4 and IL-10), was observed in the pancreas of CTB-Pins-treated NOD mice. Serum levels of immunoglobulin G1 (IgG1), but not IgG2a, were elevated in CTB-Pins-treated mice. Taken together, T-helper 2 (Th2) lymphocyte-mediated oral tolerance is a likely mechanism for the prevention of pancreatic insulitis and the preservation of insulin-producing β-cells. This is the first report of expression of a therapeutic protein in transgenic chloroplasts of an edible crop. Transplastomic lettuce plants expressing CTB-Pins grew normally and transgenes were maternally inherited in T1 progeny. This opens up the possibility for the low-cost production and delivery of human therapeutic proteins, and a strategy for the treatment of various other autoimmune diseases. PMID:17490448

  10. Expression and characterization of recombinant human serum ...

    African Journals Online (AJOL)

    C-peptide (CP), connecting the A and B chains in proinsulin, has been considered to possess physiological effects in diabetes. In order to prolong the half-life of CP in vivo, a long acting CP analog [human serum albumin (HSA-CP)] was obtained by direct gene fusion of a single-chain CP to HSA and expressed in host ...

  11. Expression and characterization of recombinant human serum ...

    African Journals Online (AJOL)

    ajl yemi

    2011-11-14

    Nov 14, 2011 ... C-peptide (CP), connecting the A and B chains in proinsulin, has been considered to possess physiological effects in diabetes. In order to prolong the half-life of CP in vivo, a long acting CP analog. [human serum albumin (HSA-CP)] was obtained by direct gene fusion of a single-chain CP to HSA and.

  12. Purification and characterization of insulin and the C-peptide of proinsulin from Przewalski's horse, zebra, rhino, and tapir (Perissodactyla).

    Science.gov (United States)

    Henry, J S; Lance, V A; Conlon, J M

    1993-02-01

    Within the order Perissodactyla, the primary structure of insulin has been strongly conserved. Insulin from Przewalski's horse and the mountain zebra (suborder Hippomorpha) is the same as that from the domestic horse and differs from insulin from the white rhinoceros and mountain tapir (suborder Ceratomorpha) by a single substitution (Gly-->Ser) at position 9 in the A-chain. A second molecular form of Przewalski's horse insulin isolated in this study was shown to represent the gamma-ethyl ester of the Glu17 residue of the A-chain. This component was probably formed during the extraction of the pancreas with acidified ethanol. The amino acid sequence of the C-peptide of proinsulin has been less well conserved. Zebra C-peptide comprises 31 amino acid residues and differs from Przewalski's horse and domestic horse C-peptide by one substitution (Gln30-->Pro). Rhino C-peptide was isolated only in a truncated form corresponding to residues (1-23) of intact C-peptide. Its amino acid sequence contains three substitutions compared with the corresponding region of horse C-peptide. It is postulated that the substitution (Pro23-->Thr) renders rhino C-peptide more liable to proteolytic cleavage by a chymotrypsin-like enzyme than horse C-peptide. C-peptide could not be identified in the extract of tapir pancreas, suggesting that proteolytic degradation may have been more extensive than in the rhino. In contrast to the ox and pig (order Artiodactyla), there was no evidence for the expression of more than one proinsulin gene in the species of Perissodactyla examined.

  13. Determination of human muscle protein fractional synthesis rate

    DEFF Research Database (Denmark)

    Bornø, Andreas; Hulston, Carl J; van Hall, Gerrit

    2014-01-01

    In the present study, different MS methods for the determination of human muscle protein fractional synthesis rate (FSR) using [ring-(13)C6 ]phenylalanine as a tracer were evaluated. Because the turnover rate of human skeletal muscle is slow, only minute quantities of the stable isotopically...

  14. Globalization, human rights, and the social determinants of health.

    Science.gov (United States)

    Chapman, Audrey R

    2009-02-01

    Globalization, a process characterized by the growing interdependence of the world's people, impacts health systems and the social determinants of health in ways that are detrimental to health equity. In a world in which there are few countervailing normative and policy approaches to the dominant neoliberal regime underpinning globalization, the human rights paradigm constitutes a widely shared foundation for challenging globalization's effects. The substantive rights enumerated in human rights instruments include the right to the highest attainable level of physical and mental health and others that are relevant to the determinants of health. The rights stipulated in these documents impose extensive legal obligations on states that have ratified these documents and confer health entitlements on their residents. Human rights norms have also inspired civil society efforts to improve access to essential medicines and medical services, particularly for HIV/AIDS. Nevertheless, many factors reduce the potential counterweight human rights might exert, including and specifically the nature of the human rights approach, weak political commitments to promoting and protecting health rights on the part of some states and their lack of institutional and economic resources to do so. Global economic markets and the relative power of global economic institutions are also shrinking national policy space. This article reviews the potential contributions and limitations of human rights to achieving greater equity in shaping the social determinants of health.

  15. Determination of positions of optical elements of the human eye

    International Nuclear Information System (INIS)

    Galetskii, S O; Cherezova, T Yu

    2009-01-01

    An original method for noninvasive determining the positions of elements of intraocular optics is proposed. The analytic dependence of the measurement error on the optical-scheme parameters and the restriction in distance from the element being measured are determined within the framework of the method proposed. It is shown that the method can be efficiently used for determining the position of elements in the classical Gullstrand eye model and personalised eye models. The positions of six optical surfaces of the Gullstrand eye model and four optical surfaces of the personalised eye model can be determined with an error of less than 0.25 mm. (human eye optics)

  16. Determination of the therapeutic potential of human umbilical cord ...

    African Journals Online (AJOL)

    This research was conducted to evaluate the therapeutic potential of human umbilical cord blood, by determining their effect on bacterial pathogens which included: Streptobacillus sp, Corynebacterium diphtheriae, Staphylococcus aureus, Salmonella typhimurium, and Escherichia coli. Cord blood samples were obtained ...

  17. Determination of atomic number and composition of human enamel

    International Nuclear Information System (INIS)

    Nogueira, M.S.; Rodas Duran, J.E.

    2001-01-01

    The teeth are organs of complicated structure that consist, partly, of hard tissue containing in its interior the dental pulp, rich in vases and nerves. The main mass of the tooth is constituted by the dentine, which is covered with hard tissues and of epithelial origin called enamel. The dentine of the human teeth used in this work were completely removed and the teeth were cut with a device with a diamond disc. In this work the chemical composition of the human enamel was determined, which showed a high percentage of Ca and P, in agreement with the results found in the literature. The effective atomic number of the material and the half-value layer in the energy range of diagnostic X-ray beams were determined. Teeth could be used to evaluated the public's individual doses as well as for retrospective dosimetry what confirms the importance of their effective atomic number and composition determination. (author)

  18. Ecological determinants of health: food and environment on human health.

    Science.gov (United States)

    Li, Alice M L

    2017-04-01

    Human health and diseases are determined by many complex factors. Health threats from the human-animal-ecosystems interface (HAEI) and zoonotic diseases (zoonoses) impose an increasing risk continuously to public health, from those emerging pathogens transmitted through contact with animals, food, water and contaminated environments. Immense challenges forced on the ecological perspectives on food and the eco-environments, including aquaculture, agriculture and the entire food systems. Impacts of food and eco-environments on human health will be examined amongst the importance of human interventions for intended purposes in lowering the adverse effects on the biodiversity. The complexity of relevant conditions defined as factors contributing to the ecological determinants of health will be illuminated from different perspectives based on concepts, citations, examples and models, in conjunction with harmful consequential effects of human-induced disturbances to our environments and food systems, together with the burdens from ecosystem disruption, environmental hazards and loss of ecosystem functions. The eco-health literacy should be further promoting under the "One Health" vision, with "One World" concept under Ecological Public Health Model for sustaining our environments and the planet earth for all beings, which is coincidentally echoing Confucian's theory for the environmental ethics of ecological harmony.

  19. Intrinsic factor in human amniotic fluid as determined by radioimmunoassay

    International Nuclear Information System (INIS)

    Wahlstedt, V.; Stenman, U.-H.; Ylinen, K.; Graesbeck, R.

    1983-01-01

    The intrinsic factor (IF) concentration in 55 human amniotic fluid specimens was determined by radioimmunoassay (RIA). The antiserum was produced by immunizing rabbits with the cobalamin-IF complex isolated from human gastric juice. The median concentration of IF was 0.17 nmol/l and the extreme values <0.07-2.51 nmol/l. Three specimens with a clearly elevated level (0.96, 1.11 and 2.51 nmol/l) were observed. The highest value was associated with a fetal malformation, viz. obstruction of the proximal gut. There was no evident correlation between the concentration of IF in amniotic fluid and gestational age. (author)

  20. Pancreatic elastase in human serum. Determination by radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Geokas, M.C. (Univ. of California, Davis); Brodrick, J.W.; Johnson, J.H.; Largman, C.

    1977-01-10

    This study demonstrates that a serine endopeptidase of pancreatic origin (elastase 2) circulates in human blood. A specific and highly sensitive radioimmunoassay has been developed for pancreatic elastase 2 in human serum. The inactivation of elastase 2 employed as radioiodinated tracer with an active site-specific reagent (phenylmethanesulfonyl fluoride) was necessary to prevent its binding by serum ..cap alpha../sub 1/-antitrypsin and ..cap alpha../sub 2/-macroglobulin while maintaining its immunoreactivity. The assay is based upon competition of standard human pancreatic elastase 2 with /sup 125/I-labeled phenylmethanesulfonyl elastase 2 for specific antibody binding sites, after which a second antibody precipitation step is used to separate bound from free /sup 125/I-labeled phenylmethanesulfonyl elastase 2. The minimum detectable concentration of elastase 2 was 0.9 ng/ml. The average normal fasting serum level determined was 71 ng/ml, approximately 80-fold greater than the minimum detectable amount.

  1. Human vision is determined based on information theory

    Science.gov (United States)

    Delgado-Bonal, Alfonso; Martín-Torres, Javier

    2016-11-01

    It is commonly accepted that the evolution of the human eye has been driven by the maximum intensity of the radiation emitted by the Sun. However, the interpretation of the surrounding environment is constrained not only by the amount of energy received but also by the information content of the radiation. Information is related to entropy rather than energy. The human brain follows Bayesian statistical inference for the interpretation of visual space. The maximization of information occurs in the process of maximizing the entropy. Here, we show that the photopic and scotopic vision absorption peaks in humans are determined not only by the intensity but also by the entropy of radiation. We suggest that through the course of evolution, the human eye has not adapted only to the maximum intensity or to the maximum information but to the optimal wavelength for obtaining information. On Earth, the optimal wavelengths for photopic and scotopic vision are 555 nm and 508 nm, respectively, as inferred experimentally. These optimal wavelengths are determined by the temperature of the star (in this case, the Sun) and by the atmospheric composition.

  2. Determination of some variables of human dentin for dosimetric purposes

    International Nuclear Information System (INIS)

    Peres, C.

    1990-01-01

    The initial objective of this work is to determine the atomic composition of human dentin, using various techniques of measurements such as X-Ray fluorescence, atomic absorption, spectrophotometry, volumetric analysis, chemistry, colorimetry and thermogravimetry. With the values of the effective atomic number Z bar dependent of the quality of the ionizing radiation, one can find the kind of material that can be used for a phantom equivalent to dentin. (author)

  3. Can we use human judgments to determine the discount rate?

    Science.gov (United States)

    Baron, J

    2000-12-01

    It has been suggested that the long-term discount rate for environmental goods should decrease at longer delays. One justification for this suggestion is that human judgments support it. This article presents an experiment showing that judgments concerning discount rates are internally inconsistent. These results point to potential problems with the use of judgments referenda for determining discount rates in cost-benefit analyses.

  4. From bacterial to human dihydrouridine synthase: automated structure determination

    Energy Technology Data Exchange (ETDEWEB)

    Whelan, Fiona, E-mail: fiona.whelan@york.ac.uk; Jenkins, Huw T., E-mail: fiona.whelan@york.ac.uk [The University of York, Heslington, York YO10 5DD (United Kingdom); Griffiths, Samuel C. [University of Oxford, Headington, Oxford OX3 7BN (United Kingdom); Byrne, Robert T. [Ludwig-Maximilians-University Munich, Feodor-Lynen-Strasse 25, 81377 Munich (Germany); Dodson, Eleanor J.; Antson, Alfred A., E-mail: fiona.whelan@york.ac.uk [The University of York, Heslington, York YO10 5DD (United Kingdom)

    2015-06-30

    The crystal structure of a human dihydrouridine synthase, an enzyme associated with lung cancer, with 18% sequence identity to a T. maritima enzyme, has been determined at 1.9 Å resolution by molecular replacement after extensive molecular remodelling of the template. The reduction of uridine to dihydrouridine at specific positions in tRNA is catalysed by dihydrouridine synthase (Dus) enzymes. Increased expression of human dihydrouridine synthase 2 (hDus2) has been linked to pulmonary carcinogenesis, while its knockdown decreased cancer cell line viability, suggesting that it may serve as a valuable target for therapeutic intervention. Here, the X-ray crystal structure of a construct of hDus2 encompassing the catalytic and tRNA-recognition domains (residues 1–340) determined at 1.9 Å resolution is presented. It is shown that the structure can be determined automatically by phenix.mr-rosetta starting from a bacterial Dus enzyme with only 18% sequence identity and a significantly divergent structure. The overall fold of the human Dus2 is similar to that of bacterial enzymes, but has a larger recognition domain and a unique three-stranded antiparallel β-sheet insertion into the catalytic domain that packs next to the recognition domain, contributing to domain–domain interactions. The structure may inform the development of novel therapeutic approaches in the fight against lung cancer.

  5. From bacterial to human dihydrouridine synthase: automated structure determination

    International Nuclear Information System (INIS)

    Whelan, Fiona; Jenkins, Huw T.; Griffiths, Samuel C.; Byrne, Robert T.; Dodson, Eleanor J.; Antson, Alfred A.

    2015-01-01

    The crystal structure of a human dihydrouridine synthase, an enzyme associated with lung cancer, with 18% sequence identity to a T. maritima enzyme, has been determined at 1.9 Å resolution by molecular replacement after extensive molecular remodelling of the template. The reduction of uridine to dihydrouridine at specific positions in tRNA is catalysed by dihydrouridine synthase (Dus) enzymes. Increased expression of human dihydrouridine synthase 2 (hDus2) has been linked to pulmonary carcinogenesis, while its knockdown decreased cancer cell line viability, suggesting that it may serve as a valuable target for therapeutic intervention. Here, the X-ray crystal structure of a construct of hDus2 encompassing the catalytic and tRNA-recognition domains (residues 1–340) determined at 1.9 Å resolution is presented. It is shown that the structure can be determined automatically by phenix.mr-rosetta starting from a bacterial Dus enzyme with only 18% sequence identity and a significantly divergent structure. The overall fold of the human Dus2 is similar to that of bacterial enzymes, but has a larger recognition domain and a unique three-stranded antiparallel β-sheet insertion into the catalytic domain that packs next to the recognition domain, contributing to domain–domain interactions. The structure may inform the development of novel therapeutic approaches in the fight against lung cancer

  6. Partial diversion of a mutant proinsulin (B10 aspartic acid) from the regulated to the constitutive secretory pathway in transfected AtT-20 cells.

    OpenAIRE

    Gross, D J; Halban, P A; Kahn, C R; Weir, G C; Villa-Komaroff, L

    1989-01-01

    A patient with type II diabetes associated with hyperproinsulinemia has been shown to have a point mutation in one insulin gene allele, resulting in replacement of histidine with aspartic acid at position 10 of the B-chain. To investigate the basis of the proinsulin processing defect, we introduced an identical mutation in the rat insulin II gene and expressed both the normal and the mutant genes in the AtT-20 pituitary corticotroph cell line. Cells expressing the mutant gene showed increased...

  7. Elevations in the Fasting Serum Proinsulin-to-C-Peptide Ratio Precede the Onset of Type 1 Diabetes.

    Science.gov (United States)

    Sims, Emily K; Chaudhry, Zunaira; Watkins, Renecia; Syed, Farooq; Blum, Janice; Ouyang, Fangqian; Perkins, Susan M; Mirmira, Raghavendra G; Sosenko, Jay; DiMeglio, Linda A; Evans-Molina, Carmella

    2016-09-01

    We tested whether an elevation in the serum proinsulin-to-C-peptide ratio (PI:C), a biomarker of β-cell endoplasmic reticulum (ER) dysfunction, was associated with progression to type 1 diabetes. Fasting total PI and C levels were measured in banked serum samples obtained from TrialNet Pathway to Prevention (PTP) participants, a cohort of autoantibody-positive relatives without diabetes of individuals with type 1 diabetes. Samples were obtained ∼12 months before diabetes onset from PTP progressors in whom diabetes developed (n = 60), and were compared with age-, sex-, and BMI-matched nonprogressors who remained normoglycemic (n = 58). PI:C ratios were calculated as molar ratios and were multiplied by 100% to obtain PI levels as a percentage of C levels. Although absolute PI levels did not differ between groups, PI:C ratios were significantly increased in antibody-positive subjects in whom there was progression to diabetes compared with nonprogressors (median 1.81% vs. 1.17%, P = 0.03). The difference between groups was most pronounced in subjects who were ≤10 years old, where the median progressor PI:C ratio was nearly triple that of nonprogressors; 90.0% of subjects in this age group within the upper PI:C quartile progressed to the development of diabetes. Logistic regression analysis, adjusted for age and BMI, demonstrated increased odds of progression for higher natural log PI:C ratio values (odds ratio 1.44, 95% CI 1.02, 2.05). These data suggest that β-cell ER dysfunction precedes type 1 diabetes onset, especially in younger children. Elevations in the serum PI:C ratio may have utility in predicting the onset of type 1 diabetes in the presymptomatic phase. © 2016 by the American Diabetes Association.

  8. Key Determinants of Human Resource Management in Hospitals: Stakeholder Perspective

    Directory of Open Access Journals (Sweden)

    Buchelt Beata Irma

    2017-06-01

    Full Text Available Over the past decade, theoretical and empirical research on the various aspects of human resources (HR within the healthcare (HC sector has grown extensively due to its′ strategic importance in the sector. There is a visible tendency among researchers to pursue an effective human resource management (HRM strategies, methods, and tools. Countries implement policies which should increase the amount and competences of employees within healthcare. Providers of HC services (i.e. hospitals tend to enforce modern HRM solutions adapted from business organisations to attract, retain and develop HR. However, these seem not be as effective as they could (Hyde et al., 2013. Because of this, authors approached a researched reality from the point of view of a contextual paradigm, assuming that HRM solutions to be effective should match the reality of HC providers (Pocztowski, 2008. The aim of the research was to detect determinants which might influence the management of medical personnel in hospitals and identify the possible strength of these determinants so a more adjusted organisational and human resource management strategy could be elaborated. The list of possible determinants of hospital operations as the result of meta-analysis was elaborated. The list created the basis for interviews conducted among stakeholders and experts. Respondents were asked to appraise the factors with the usage of numerical scale considering their influence on medical personnel management in hospitals (physicians, nurses and others. In total, there were 28 interviews completed. The general conclusion which can be drawn from the analysis of these data is that hospitals should reorient their HRM practices in such the way that not only the quantitative but also the qualitative aspect of performance would be properly handled. This paper draws from HRM theory (contextual approach, stakeholder theory, and healthcare management theory, adding new insight to each in the context of

  9. Radioimmunoassay for determination of thyroglobulin in human serum

    International Nuclear Information System (INIS)

    Rodriguez Cabrera, M.E.; Blanca Fernandez, S.; Baldor Navarro, F.; Rodriguez Gonzalez, J.C.

    1996-01-01

    We described the development and analytical evaluation of a radioimmunoassay with double antibody in liquid phase for human serum thyroglobulin determination using a set of reagents produced in the National Institute of Endocrinology. The reference interval for normal population was 5.7 - 44 ng/ml (X± 2DS; n=170). The intra-assay precipision was 8.1 % (control serum A) and 7.0 (control serum B) and the inter-assay precision was 9.1 % (control serum A) and 9.2 % (control serum B). The sensibility was 4.7 ng/ml and the recovery was 95 %

  10. Determinants of human resource investment in internal controls

    Directory of Open Access Journals (Sweden)

    Jong-Hag Choi

    2013-09-01

    Full Text Available Using the unique reporting environment in Korea, this study investigates the determinants of human resource investment in internal controls for 1352 listed firms disclosing the number of personnel who are in charge of internal control-related tasks (IC personnel from 2005 to 2008. We find that the number of IC personnel within a firm and several key departments increase with firm size, number of employees, complexity and for Chaebols, and decrease in rapidly growing firms. Additional analysis reveals that the factors influencing internal control systems have an accentuated effect on firms with relatively larger firm size.

  11. Purification and primary structure determination of human lysosomal dipeptidase.

    Science.gov (United States)

    Dolenc, Iztok; Mihelic, Marko

    2003-02-01

    The lysosomal metallopeptidase is an enzyme that acts preferentially on dipeptides with unsubstituted N- and C-termini. Its activity is highest in slightly acidic pH. Here we describe the isolation and characterization of lysosomal dipeptidase from human kidney. The isolated enzyme has the amino-terminal sequence DVAKAIINLAVY and is a homodimer with a molecular mass of 100 kDa. So far no amino acid sequence has been determined for this metallopeptidase. The complete primary structure as deduced from the nucleotide sequence revealed that the isolated dipeptidase is similar to blood plasma glutamate carboxypeptidase.

  12. Automation of gender determination in human canines using artificial intelligence

    Directory of Open Access Journals (Sweden)

    F. Fidya

    2017-09-01

    Full Text Available Background: Gender determination is an important aspect of the identification process. The tooth represents a part of the human body that indicates the nature of sexual dimorphism. Artificial intelligence enables computers to perform to the same standard the same tasks as those carried out by humans. Several methods of classification exist within an artificial intelligence approach to identifying sexual dimorphism in canines. Purpose: This study aimed to quantify the respective accuracy of the Naive Bayes, decision tree, and multi-layer perceptron (MLP methods in identifying sexual dimorphism in canines. Methods: A sample of results derived from 100 measurements of the diameter of mesiodistal, buccolingual, and diagonal upper and lower canine jaw models of both genders were entered into an application computer program that implements the algorithm (MLP. The analytical process was conducted by the program to obtain a classification model with testing being subsequently carried out in order to obtain 50 new measurement results, 25 each for males and females. A comparative analysis was conducted on the program-generated information. Results: The accuracy rate of the Naive Bayes method was 82%, while that of the decision tree and MLP amounted to 84%. The MLP method had an absolute error value lower than that of its decision tree counterpart. Conclusion: The use of artificial intelligence methods produced a highly accurate identification process relating to the gender determination of canine teeth. The most appropriate method was the MLP with an accuracy rate of 84%.

  13. Determination of trace elements in the human substantia nigra

    Energy Technology Data Exchange (ETDEWEB)

    Morawski, M. [Paul-Flechsig-Institut fuer Hirnforschung, Universitaet Leipzig, Jahnallee 59, 04109 Leipzig (Germany)]. E-mail: morm@medizin.uni-leipzig.de; Meinecke, Ch. [Fakultaet fuer Physik und Geowissenschaften, Universitaet Leipzig, Linnestrasse 5, 04105 Leipzig (Germany); Reinert, T. [Fakultaet fuer Physik und Geowissenschaften, Universitaet Leipzig, Linnestrasse 5, 04105 Leipzig (Germany); Doerffel, A.C. [Paul-Flechsig-Institut fuer Hirnforschung, Universitaet Leipzig, Jahnallee 59, 04109 Leipzig (Germany); Riederer, P. [Klin. Neurochemie, Abt. Psychiatrie, Universitaet Wuerzburg, Fuechsleinstrasse 15, 97080 Wuerzburg (Germany); Arendt, T. [Paul-Flechsig-Institut fuer Hirnforschung, Universitaet Leipzig, Jahnallee 59, 04109 Leipzig (Germany); Butz, T. [Fakultaet fuer Physik und Geowissenschaften, Universitaet Leipzig, Linnestrasse 5, 04105 Leipzig (Germany)

    2005-04-01

    'The gain in brain is mainly in the stain' was long time a key sentence for research in neurodegenerative disease. However, for a quantification of the element concentrations (especially iron) in brain tissue, standard staining methods are insufficient. Advanced physical methods allow a quantitative elemental analysis of brain tissue. The sophisticated ion beam analysis provides a quantitative determination of elemental concentrations with a subcellular spatial resolution using a scanning proton beam focussed down to below 1 {mu}m that induces characteristic X-rays in the specimen (PIXE - particle induced X-ray emission). Histochemical and biochemical determinations of total iron content in brain regions from idiopathic Parkinson's disease have demonstrated an increase of iron in parkinsonian substantia nigra pars compacta but not in the pars reticulata, however without a clear cellular classification. For the first time, we have differentially investigated the intra- and extraneuronal elemental concentrations (especially iron) of the human substantia nigra pars compacta versus pars reticulata with detection limits in the range of 50 {mu}mol/l. Thus, we could compare the neuronal iron concentration in human brain sections of healthy and parkinsonian brain tissue. Clear differences in the iron concentration and distribution could be disclosed. Additionally, we could show in situ that the increased intraneuronal iron content is linked to neuromelanin.

  14. New validated method for piracetam HPLC determination in human plasma.

    Science.gov (United States)

    Curticapean, Augustin; Imre, Silvia

    2007-01-10

    The new method for HPLC determination of piracetam in human plasma was developed and validated by a new approach. The simple determination by UV detection was performed on supernatant, obtained from plasma, after proteins precipitation with perchloric acid. The chromatographic separation of piracetam under a gradient elution was achieved at room temperature with a RP-18 LiChroSpher 100 column and aqueous mobile phase containing acetonitrile and methanol. The quantitative determination of piracetam was performed at 200 nm with a lower limit of quantification LLQ=2 microg/ml. For this limit, the calculated values of the coefficient of variation and difference between mean and the nominal concentration are CV%=9.7 and bias%=0.9 for the intra-day assay, and CV%=19.1 and bias%=-7.45 for the between-days assay. For precision, the range was CV%=1.8/11.6 in the intra-day and between-days assay, and for accuracy, the range was bias%=2.3/14.9 in the intra-day and between-days assay. In addition, the stability of piracetam in different conditions was verified. Piracetam proved to be stable in plasma during 4 weeks at -20 degrees C and for 36 h at 20 degrees C in the supernatant after protein precipitation. The new proposed method was used for a bioequivalence study of two medicines containing 800 mg piracetam.

  15. The determinants of alternative RNA splicing in human cells.

    Science.gov (United States)

    Ramanouskaya, Tatsiana V; Grinev, Vasily V

    2017-12-01

    Alternative splicing represents an important level of the regulation of gene function in eukaryotic organisms. It plays a critical role in virtually every biological process within an organism, including regulation of cell division and cell death, differentiation of tissues in the embryo and the adult organism, as well as in cellular response to diverse environmental factors. In turn, studies of the last decade have shown that alternative splicing itself is controlled by different mechanisms. Unfortunately, there is no clear understanding of how these diverse mechanisms, or determinants, regulate and constrain the set of alternative RNA species produced from any particular gene in every cell of the human body. Here, we provide a consolidated overview of alternative splicing determinants including RNA-protein interactions, epigenetic regulation via chromatin remodeling, coupling of transcription-to-alternative splicing, effect of secondary structures in pre-RNA, and function of the RNA quality control systems. We also extensively and critically discuss some mechanistic insights on coordinated inclusion/exclusion of exons during the formation of mature RNA molecules. We conclude that the final structure of RNA is pre-determined by a complex interplay between cis- and trans-acting factors. Altogether, currently available empirical data significantly expand our understanding of the functioning of the alternative splicing machinery of cells in normal and pathological conditions. On the other hand, there are still many blind spots that require further deep investigations.

  16. Determination of pyrazinamide in human by high performance liquid chromatography.

    Directory of Open Access Journals (Sweden)

    Revankar S

    1994-01-01

    Full Text Available A facile and sensitive high performance liquid chromatographic (HPLC technique has been developed for the determination pyrazinamide (PZA in human plasma. Nicotinamide(NIA is used as internal standard(IS. Plasma is deproteinized with 0.7 M perchloric acid; clear supernatant is neutralized with 1M NaOH and injected onto HPLC. The separation of pyrazinamide and the internal standard is carried out on a Supelco LC-18 (DB column with a basic mobile phase. Pyrazinoic acid, the major metabolite, other anti-tuberculous drugs and endogenous components do not interfere with measurement of pyrazinamide. The limit of detection of pyrazinamide with this method is 0.2 mg/0.2 ml plasma (CV 8.2%.

  17. Variation in human recombination rates and its genetic determinants.

    Directory of Open Access Journals (Sweden)

    Adi Fledel-Alon

    Full Text Available Despite the fundamental role of crossing-over in the pairing and segregation of chromosomes during human meiosis, the rates and placements of events vary markedly among individuals. Characterizing this variation and identifying its determinants are essential steps in our understanding of the human recombination process and its evolution.Using three large sets of European-American pedigrees, we examined variation in five recombination phenotypes that capture distinct aspects of crossing-over patterns. We found that the mean recombination rate in males and females and the historical hotspot usage are significantly heritable and are uncorrelated with one another. We then conducted a genome-wide association study in order to identify loci that influence them. We replicated associations of RNF212 with the mean rate in males and in females as well as the association of Inversion 17q21.31 with the female mean rate. We also replicated the association of PRDM9 with historical hotspot usage, finding that it explains most of the genetic variance in this phenotype. In addition, we identified a set of new candidate regions for further validation.These findings suggest that variation at broad and fine scales is largely separable and that, beyond three known loci, there is no evidence for common variation with large effects on recombination phenotypes.

  18. Determinants of Human African Trypanosomiasis Elimination via Paratransgenesis.

    Directory of Open Access Journals (Sweden)

    Jennifer A Gilbert

    2016-03-01

    Full Text Available Human African trypanosomiasis (HAT, transmitted by tsetse flies, has historically infected hundreds of thousands of individuals annually in sub-Saharan Africa. Over the last decade, concerted control efforts have reduced reported cases to below 10,000 annually, bringing complete elimination within reach. A potential technology to eliminate HAT involves rendering the flies resistant to trypanosome infection. This approach can be achieved through the introduction of transgenic Sodalis symbiotic bacteria that have been modified to produce a trypanocide, and propagated via Wolbachia symbionts, which confer a reproductive advantage to the paratransgenic tsetse. However, the population dynamics of these symbionts within tsetse flies have not yet been evaluated. Specifically, the key factors that determine the effectiveness of paratransgenesis have yet to be quantified. To identify the impact of these determinants on T.b. gambiense and T.b. rhodesiense transmission, we developed a mathematical model of trypanosome transmission that incorporates tsetse and symbiont population dynamics. We found that fecundity and mortality penalties associated with Wolbachia or recombinant Sodalis colonization, probabilities of vertical transmission, and tsetse migration rates are fundamental to the feasibility of HAT elimination. For example, we determined that HAT elimination could be sustained over 25 years when Wolbachia colonization minimally impacted fecundity or mortality, and when the probability of recombinant Sodalis vertical transmission exceeded 99.9%. We also found that for a narrow range of recombinant Sodalis vertical transmission probability (99.9-90.6% for T.b. gambiense and 99.9-85.8% for T.b. rhodesiense, cumulative HAT incidence was reduced between 30% and 1% for T.b. gambiense and between 21% and 3% for T.b. rhodesiense, although elimination was not predicted. Our findings indicate that fitness and mortality penalties associated with paratransgenic

  19. Production of peptide antisera specific for mouse and rat proinsulin C-peptide 2

    DEFF Research Database (Denmark)

    Blume, N; Madsen, O D; Kofod, Hans

    1990-01-01

    for antibody binding to the immunizing antigen. Antisera to C-peptide 2, stained islet beta-cells on mouse and rat, but not monkey pancreas sections in immunocytochemical analysis. Preabsorption to the synthetic C-peptide 2, but not the synthetic mouse and rat C-peptide 1 abolished staining. In conclusion we......Mice and rats have two functional non-allelic insulin genes. By using a synthetic peptide representing a common sequence in mouse and rat C-peptide 2 as antigen, we have produced rabbit antisera specific for an epitope which is not present in mouse or rat C-peptide 1. Long-term immunization did...... not seem to increase the end point titre as tested in direct ELISA. The specificity of the antiserum was determined by competitive ELISA and histochemistry on pancreas sections. Only the synthetic C-peptide 2, but not the homologous synthetic C-peptide 1 from mouse and rat competed efficiently in ELISA...

  20. Institutionalising Human Governance Determinant: Steering Organizations towards Sustainability

    Science.gov (United States)

    Muhamad Hanapiyah, Zulkefli; Daud, Salina; Abdullah, Wan Mohammad Taufik Wan

    2016-03-01

    This paper discusses past researches on human governance elements. Eight elements of human governance are proposed in this paper: leadership, integrity, religiosity, spirituality, culture, recruitment, training and internal control policy. Empirical study shall be conducted in the future study to confirm the eight elements of human governance proposed in this paper. It is expected that these elements may enhance the human governance practice in the organizations.

  1. Modeling for Determinants of Human Trafficking: An Empirical Analysis

    Directory of Open Access Journals (Sweden)

    Seo-Young Cho

    2015-02-01

    Full Text Available This study aims to identify robust push and pull factors of human trafficking. I test for the robustness of 70 push and 63 pull factors suggested in the literature. In doing so, I employ an extreme bound analysis, running more than two million regressions with all possible combinations of variables for up to 153 countries during the period of 1995–2010. My results show that crime prevalence robustly explains human trafficking both in destination and origin countries. Income level also has a robust impact, suggesting that the cause of human trafficking shares that of economic migration. Law enforcement matters more in origin countries than destination countries. Interestingly, a very low level of gender equality may have constraining effects on human trafficking outflow, possibly because gender discrimination limits female mobility that is necessary for the occurrence of human trafficking.

  2. Genetical genomic determinants of alcohol consumption in rats and humans

    Directory of Open Access Journals (Sweden)

    Mangion Jonathan

    2009-10-01

    Full Text Available Abstract Background We have used a genetical genomic approach, in conjunction with phenotypic analysis of alcohol consumption, to identify candidate genes that predispose to varying levels of alcohol intake by HXB/BXH recombinant inbred rat strains. In addition, in two populations of humans, we assessed genetic polymorphisms associated with alcohol consumption using a custom genotyping array for 1,350 single nucleotide polymorphisms (SNPs. Our goal was to ascertain whether our approach, which relies on statistical and informatics techniques, and non-human animal models of alcohol drinking behavior, could inform interpretation of genetic association studies with human populations. Results In the HXB/BXH recombinant inbred (RI rats, correlation analysis of brain gene expression levels with alcohol consumption in a two-bottle choice paradigm, and filtering based on behavioral and gene expression quantitative trait locus (QTL analyses, generated a list of candidate genes. A literature-based, functional analysis of the interactions of the products of these candidate genes defined pathways linked to presynaptic GABA release, activation of dopamine neurons, and postsynaptic GABA receptor trafficking, in brain regions including the hypothalamus, ventral tegmentum and amygdala. The analysis also implicated energy metabolism and caloric intake control as potential influences on alcohol consumption by the recombinant inbred rats. In the human populations, polymorphisms in genes associated with GABA synthesis and GABA receptors, as well as genes related to dopaminergic transmission, were associated with alcohol consumption. Conclusion Our results emphasize the importance of the signaling pathways identified using the non-human animal models, rather than single gene products, in identifying factors responsible for complex traits such as alcohol consumption. The results suggest cross-species similarities in pathways that influence predisposition to consume

  3. HPLC Method for Determination of Rifaximin in Human Plasma ...

    African Journals Online (AJOL)

    The present study was aimed at developing a simple, sensitive, and specific liquid chromatography–tandem mass spectrometry method for the quantification of rifaximin in human plasma using rifaximin D6 as internal standard. Chromatographic separation was performed on Zorbax SB C18, 4.6 x 75 mm, 3.5 μm column with ...

  4. Determination of PBDEs in human milk. Analysis and quality control

    Energy Technology Data Exchange (ETDEWEB)

    Paepke, O.; Herrmann, T. [Ergo Research, Hamburg (Germany); Vieth, B.; Ostermann, B. [Federal Institute for Risk Assessment, Berlin (Germany)

    2004-09-15

    Polybrominated diphenylethers (PBDEs) are widely used as flame retardants in polymer materials, textiles, electronic boards and various other materials. Technical PBDE preparations are produced as mixtures of mainly penta-, octa- or decabromobiphenyl ethers. PBDEs are structurally similar to other environmental pollutants, such as dioxins and PCBs. They are lipophilic and persistent compounds and widespread in the environment. For certain congeners, bioaccumulation has been observed. Recent findings of increasing levels in humans showed that more detailed investigations of human milk (or other suitable matrices) will be required in order to evaluate the general human exposure to this group of environmental contaminants. Only a few data on PBDE levels in breast milk from Germany had been published. To fill the data gaps, in 2001 a controlled study was started to characterize the PBDE levels in human milk from Germany with special efforts to identify and quantify deca-BDE-209. 103 samples were analyzed in this study so far including 10 hidden pool samples provided by Federal Institute for Risk Assessment to ERGO laboratory (total number of samples finally will be 157 (including 14 hidden pool samples)). This paper describes the analytical procedure applied and emphasizes on the quality control procedure.

  5. Human physiology as the determining factor in protective clothing design

    NARCIS (Netherlands)

    Daanen, Hein

    2014-01-01

    Protective clothing is designed to protect humans against risks like fire, chemicals or blunt impact. Although protect¡ve clothing diminishes the effects of external risks, it may hinder people in functioning and it may also introduce new (internal) risks. Manufacturers are often not aware of the

  6. Determination of Theophylline Binding to Human Serum Proteins by Isotachophoresis

    NARCIS (Netherlands)

    Reijenga, J.C.; Gaijkema, A.P.M.; Mikkers, F.E.P.

    1984-01-01

    Free theophylline was isolated from human serum by ultrafiltration and analysed in a leading electrolyte of 7.5 mM morpholinoethanesulphoric acid with ammediol as a counter ion at pH 8.90 and -alanine as a terminator. The UV (280 nm) absorbance of the theophylline spike between serine and bicine as

  7. Socio-economic and demographic determinants of Human ...

    African Journals Online (AJOL)

    Approaches to reduce related morbidity and mortality include screening for cervical cancer and administration of Human Papilloma Virus (HPV) vaccine to HPV non-infected females to trigger host resistance. The uptake of HPV vaccine globally has been reported to be less than 1%. Objectives: To assess the level and ...

  8. Human Papilloma Virus Vaccine: Determinants of Acceptability by ...

    African Journals Online (AJOL)

    Vaccination of adolescent females against Human Papilloma Virus (HPV), the causative agent for cervical cancer has recently become available. As minors, parental acceptance of the vaccines for adolescent daughters requires exploration. This was a cross-sectional survey of 201 mothers attending the gynaecology clinic ...

  9. Infrared analysis for determining macronutrients in human milk

    DEFF Research Database (Denmark)

    Michaelsen, K F; Pedersen, S B; Skafte, L

    1988-01-01

    Infrared (IR) analysis is widely used for routine analysis of cow milk in dairies. The aim of this study was to evaluate the precision and accuracy of an IR analyzer (Milko-scan 104) for measuring protein, fat, carbohydrate, and, indirectly, the energy content of human milk. The results of the IR...

  10. Habituation as a Determinant of Human Food Intake

    Science.gov (United States)

    Epstein, Leonard H.; Temple, Jennifer L.; Roemmich, James N.; Bouton, Mark E.

    2009-01-01

    Research has shown that animals and humans habituate on a variety of behavioral and physiological responses to repeated presentations of food cues, and habituation is related to amount of food consumed and cessation of eating. The purpose of this article is to provide an overview of experimental paradigms used to study habituation, integrate a…

  11. Comparison of Methods for Dependency Determination between Human Failure Events within Human Reliability Analysis

    International Nuclear Information System (INIS)

    Cepin, M.

    2008-01-01

    The human reliability analysis (HRA) is a highly subjective evaluation of human performance, which is an input for probabilistic safety assessment, which deals with many parameters of high uncertainty. The objective of this paper is to show that subjectivism can have a large impact on human reliability results and consequently on probabilistic safety assessment results and applications. The objective is to identify the key features, which may decrease subjectivity of human reliability analysis. Human reliability methods are compared with focus on dependency comparison between Institute Jozef Stefan human reliability analysis (IJS-HRA) and standardized plant analysis risk human reliability analysis (SPAR-H). Results show large differences in the calculated human error probabilities for the same events within the same probabilistic safety assessment, which are the consequence of subjectivity. The subjectivity can be reduced by development of more detailed guidelines for human reliability analysis with many practical examples for all steps of the process of evaluation of human performance

  12. Comparison of methods for dependency determination between human failure events within human reliability analysis

    International Nuclear Information System (INIS)

    Cepis, M.

    2007-01-01

    The Human Reliability Analysis (HRA) is a highly subjective evaluation of human performance, which is an input for probabilistic safety assessment, which deals with many parameters of high uncertainty. The objective of this paper is to show that subjectivism can have a large impact on human reliability results and consequently on probabilistic safety assessment results and applications. The objective is to identify the key features, which may decrease of subjectivity of human reliability analysis. Human reliability methods are compared with focus on dependency comparison between Institute Jozef Stefan - Human Reliability Analysis (IJS-HRA) and Standardized Plant Analysis Risk Human Reliability Analysis (SPAR-H). Results show large differences in the calculated human error probabilities for the same events within the same probabilistic safety assessment, which are the consequence of subjectivity. The subjectivity can be reduced by development of more detailed guidelines for human reliability analysis with many practical examples for all steps of the process of evaluation of human performance. (author)

  13. Enhanced Chromatographic Determination of Nicotine in Human Plasma: Applied to Human Volunteers.

    Science.gov (United States)

    Ayoub, Bassam M; Mohamady, Samy; Hendy, Moataz S; Elmazar, Mohamed M

    2015-12-01

    Development of enhanced UPLC-UV method for determination of nicotine in human plasma was achieved on a Symmetry(®) C18 column (100 mm × 2.1 mm, 2.2 μm) applying isocratic elution based on Methanol: Acetonitrile: Phosphate Buffer (pH: 2.7) with the ratio (20:30:50, v/v/v) as a mobile phase. The ultraviolet detector was operated at 260 nm. The mobile phase was pumped through the column at a flow rate of 0.2 mL min(-1). The column temperature was adjusted to 50ºC and the injection volume was 2 μL. Quinine was selected as an internal standard (IS) due to its structure similarity to nicotine having basic pyridine ring to optimize the liquid liquid extraction procedure using diethyl ether coupled with vacuum evaporation at 40°C. Validation parameters for nicotine were found to be acceptable over the concentration range of 2.5-50 ng ml(-1). The application of the proposed method on four healthy human volunteers was approved by the ethical committee. The study was carried out under fasting conditions and the concerned subjects were informed about the objectives and possible risks involved in the study. The proposed method proved to be simple and fast which is a major advantage to analyze large number of samples per day using the accelerated vacuum evaporation technique. The method showed satisfactory data for all the parameters tested within the limits for bioanalytical assays. The lower limit of quantification (LLOQ) permits the application of the method for further pharmacological and clinical studies.

  14. Determination of Sr90 activity in human bones

    International Nuclear Information System (INIS)

    Mendonca, Anamelia Habib

    1970-01-01

    Several studies have been published in the literature on the extent and levels of radioactive contamination of food chains caused by fallout from nuclear weapon tests. According to UNSCEAR, these studies cover a great number of:-areas of the developed world, though large, areas of Asia, Africa and South-America are left aside with only, unsatisfactory information about the levels, of radioactive contamination. In 1968, UNSCEAR recommended that a survey on the contamination of biological materials such as human - bone by fission products and particularly Sr 90 should be encouraged on those areas where only fragmentary information was available. UNSCEAR recommendations call upon the fact that many individuals on such areas of the world have been exposed to Sr 90 contamination from birth to their adult area. Therefore, that group have an Sr 90 skeletal burden very much different from people exposed only at adult age. Based on these considerations, UNSCEAR recommendations called for Sr 90 analysis on human bones from different age groups. In Brazil, studies on the of Sr 90 in human bone are practically non-existent, except for the year of 1959. Following UNSCEAR recommendations, we decided to perform such a survey on Sr 90 levels in human bones. Samples were collected from individuals that died in Rio de Janeiro from accidents. These samples were firstly classified according to social level in very poor and poor groups. Samples were then classified in three age groups ranging 0-18, 18-30 and 30-40 years of age. Results show that levels found in the Brazilian age groups are close to the ones observed in Chile (1969), Argentina and Australia (1966-1968) and slightly, higher than -those observed in Venezuela, Senegal and Jamaica (1969). If one compares the results obtained for the North and South hemispheres respectively, one sees that there was a more pronounced decrease in the levels of Sr 90 content of the of some regions of South America. Our results show no

  15. Determination of activities of human carbonic anhydrase II inhibitors ...

    African Journals Online (AJOL)

    Purpose: To evaluate the activities of new curcumin analogs as carbonic anhydrase II (CA-II) inhibitor. Methods: Carbonic anhydrase II (CA-II) inhibition was determined by each ligand capability to inhibit the esterase activity of CA-II using 4-NPA as a substrate in 96-well plates. Dimethyl sulfoxide was used to dissolve each ...

  16. Critical Technology Determination for Future Human Space Flight

    Science.gov (United States)

    Mercer, Carolyn R.; Vangen, Scott D.; Williams-Byrd, Julie A.; Stecklein, Jonette M.; Rahman, Shamim A.; Rosenthal, Matthew E.; Hornyak, David M.; Alexander, Leslie; Korsmeyer, David J.; Tu, Eugene L.; hide

    2012-01-01

    As the National Aeronautics and Space Administration (NASA) prepares to extend human presence throughout the solar system, technical capabilities must be developed to enable long duration flights to destinations such as near Earth asteroids, Mars, and extended stays on the Moon. As part of the NASA Human Spaceflight Architecture Team, a Technology Development Assessment Team has identified a suite of critical technologies needed to support this broad range of missions. Dialog between mission planners, vehicle developers, and technologists was used to identify a minimum but sufficient set of technologies, noting that needs are created by specific mission architecture requirements, yet specific designs are enabled by technologies. Further consideration was given to the re-use of underlying technologies to cover multiple missions to effectively use scarce resources. This suite of critical technologies is expected to provide the needed base capability to enable a variety of possible destinations and missions. This paper describes the methodology used to provide an architecture-driven technology development assessment ("technology pull"), including technology advancement needs identified by trade studies encompassing a spectrum of flight elements and destination design reference missions.

  17. Determination of human coronary artery composition by Raman spectroscopy.

    Science.gov (United States)

    Brennan, J F; Römer, T J; Lees, R S; Tercyak, A M; Kramer, J R; Feld, M S

    1997-07-01

    We present a method for in situ chemical analysis of human coronary artery using near-infrared Raman spectroscopy. It is rapid and accurate and does not require tissue removal; small volumes, approximately 1 mm3, can be sampled. This methodology is likely to be useful as a tool for intravascular diagnosis of artery disease. Human coronary artery segments were obtained from nine explanted recipient hearts within 1 hour of heart transplantation. Minces from one or more segments were obtained through grinding in a mortar and pestle containing liquid nitrogen. Artery segments and minces were excited with 830 nm near-infrared light, and Raman spectra were collected with a specially designed spectrometer. A model was developed to analyze the spectra and quantify the amounts of cholesterol, cholesterol esters, triglycerides and phospholipids, and calcium salts present. The model provided excellent fits to spectra from the artery segments, indicating its applicability to intact tissue. In addition, the minces were assayed chemically for lipid and calcium salt content, and the results were compared. The relative weights obtained using the Raman technique agreed with those of the standard assays within a few percentage points. The chemical composition of coronary artery can be quantified accurately with Raman spectroscopy. This opens the possibility of using histochemical analysis to predict acute events such as plaque rupture, to follow the progression of disease, and to select appropriate therapeutic interventions.

  18. Comparison of French and Estonian Students' Conceptions in Genetic Determinism of Human Behaviours

    Science.gov (United States)

    Castera, Jeremy; Sarapuu, Tago; Clement, Pierre

    2013-01-01

    Innatism is the belief that most of the human personality can be determined by genes. This ideology is dangerous, especially when it claims to be scientific. The present study investigates conceptions of 1060 students from Estonia and France related to genetic determinism of some human behaviours. Factors taken into account included students'…

  19. Microextraction of Furosemide from Human Serum and Its Fluorimetric Determination

    Directory of Open Access Journals (Sweden)

    Akbar Rezazadeh

    2018-03-01

    Full Text Available Background: A new, fast and sensitive spectrofluorimetric method was proposed for the determination of furosemide in serum samples based on a dispersive liquid-liquid microextraction. Methods: The optimum conditions for quantification of furosemide were studied considering the effects of types and amounts of dispersive and extraction solvents, salt addition, pH value, rate and duration of centrifugation. The method was validated with respect to the linearity, recovery and limit of detection. Results: Under the optimal conditions, the fluorescence intensities at 406 nm (with the excitation wavelength of 342 nm were linear with the concentration of furosemide in the range of 0.3 to 20 µg mL-1, with a detection limit of 0.12 µg mL-1 and a relative standard deviation of 3.4–9.4%. Conclusion: Careful examination of the obtained validation results reveal that the proposed method is suitable for determination of furosemide in serum samples.

  20. Determination of iodine in human milk and urine | Ayodele | Ife ...

    African Journals Online (AJOL)

    Physiological concentrations of iodine were determined in milk and urine. Recovery studies are reported along with results for the analysis of milk and urine samples. Iodine contents ranged from 10 - 110 (mean 52.88 ± 22.60mg/l) and 10 - 90 (mean 27.64 ±16.70) g/l in milk and urine respectively. A significant difference is ...

  1. Determination of somatic mutations in human erythrocytes by cytometry

    International Nuclear Information System (INIS)

    Jensen, R.H.; Langlois, R.G.; Bigbee, W.L.

    1985-01-01

    Flow cytometric assays of human erythrocytes labeled with monoclonal antibodies specific for glycophorin A were used to enumerate variant cells that appear in peripheral blood as a result of somatic gene-loss mutations in erythrocyte precursor cells. The assay was performed on erythrocytes from 10 oncology patients who had received at least one treatment from radiation or mutagenic chemotherapy at least 3 weeks before being assayed. The patients were suffering from many different malignancies (e.g., breast, renal, bone, colon and lung), and were treated with several different mutagenic therapeutics (e.g., cisplatinum, adriamycin, daunomycin, or cyclophosphamide). The frequency of these variant cells is an indication of the amount of mutagenic damage accumulated in the individual's erythropoietic cell population. Comparing these results to HPRT clonogenic assays, we find similar baseline frequencies of somatic mutation as well as similar correlation with mutagenic exposures. 9 refs., 3 figs., 1 tab

  2. Determination of somatic mutations in human erythrocytes by cytometry

    Energy Technology Data Exchange (ETDEWEB)

    Jensen, R.H.; Langlois, R.G.; Bigbee, W.L.

    1985-06-21

    Flow cytometric assays of human erythrocytes labeled with monoclonal antibodies specific for glycophorin A were used to enumerate variant cells that appear in peripheral blood as a result of somatic gene-loss mutations in erythrocyte precursor cells. The assay was performed on erythrocytes from 10 oncology patients who had received at least one treatment from radiation or mutagenic chemotherapy at least 3 weeks before being assayed. The patients were suffering from many different malignancies (e.g., breast, renal, bone, colon and lung), and were treated with several different mutagenic therapeutics (e.g., cisplatinum, adriamycin, daunomycin, or cyclophosphamide). The frequency of these variant cells is an indication of the amount of mutagenic damage accumulated in the individual's erythropoietic cell population. Comparing these results to HPRT clonogenic assays, we find similar baseline frequencies of somatic mutation as well as similar correlation with mutagenic exposures. 9 refs., 3 figs., 1 tab.

  3. Peripheral vasodilatation determines cardiac output in exercising humans

    DEFF Research Database (Denmark)

    Bada, A A; Svendsen, J H; Secher, N H

    2012-01-01

    In dogs, manipulation of heart rate has no effect on the exercise-induced increase in cardiac output. Whether these findings apply to humans remain uncertain, because of the large differences in cardiovascular anatomy and regulation. To investigate the role of heart rate and peripheral...... arterial ATP infusion at rest. Exercise and ATP infusion increased cardiac output, leg blood flow and vascular conductance (P heart rate by up to 54 beats min(−1), cardiac output did not change in any of the three...... demonstrate that the elevated cardiac output during steady-state exercise is regulated by the increase in skeletal muscle blood flow and venous return to the heart, whereas the increase in heart rate appears to be secondary to the regulation of cardiac output....

  4. Significance of molecular diagnostics in human papilloma virus (HPV determination

    Directory of Open Access Journals (Sweden)

    Kovačević G.

    2014-01-01

    Full Text Available HPV infection is considered to be the most important etiologic factor in cervical cancer development. In this retrospective study, which included the period from 2000 to 2012, the results of two molecular techniques used in the detection of HPV infection among women of the South Bačka District were analyzed. By using the technique of in situ hybridization and the rPCR method, the proportion of high-risk HPV among women with normal cytology was determined to be 19.8% and 32.7%, respectively, and among women with abnormal cytology 43.1% and 61%, respectively. Among the analyzed women, HPV type 16 was the most prevalent, followed by HPV types 31, 51 and 18. Application of molecular HPV diagnosis is valuable because it increases the sensitivity of the screening test, so that the application of both tests to detect cervical cancer is a true prevention of malignancy.

  5. Studies on proinsulin and proglucagon biosynthesis and conversion at the subcellular level: I. Fractionation procedure and characterization of the subcellular fractions

    Science.gov (United States)

    Noe, BD; Baste, CA; Bauer, GE

    1977-01-01

    Anglerfish islets were homogenized in 0.25 M sucrose and separated into seven separate subcellular fractions by differential and discontinuous density gradient centrifugation. The objective was to isolate microsomes and secretory granules in a highly purified state. The fractions were characterized by electron microscopy and chemical analyses. Each fraction was assayed for its content of protein, RNA, DNA, immunoreactive insulin (IRI), and immunoreactive glucagon (IRG). Ultrastructural examination showed that two of the seven subcellular fractions contain primarily mitochondria, and that two others consist almost exclusively of secretory granules. A fifth fraction contains rough and smooth microsomal vesicles. The remaining two fractions are the cell supernate and the nuclei and cell debris. The content of DNA and RNA in all fractions is consistent with the observed ultrastructure. More than 82 percent of the total cellular IRI and 89(percent) of the total cellular IRG are found in the fractions of secretory granules. The combined fractions of secretory granules and microsomes consistently yield >93 percent of the total IRG. These results indicate that the fractionation procedure employed yields fractions of microsomes and secretory granules that contain nearly all the immunoassayable insulin and glucagons found in whole islet tissue. These fractions are thus considered suitable for study of proinsulin and proglucagon biosynthesis and their metabolic conversion at the subcellular level. PMID:328517

  6. Tryptophan and kynurenine determination in human hair by liquid chromatography.

    Science.gov (United States)

    Dario, Michelli F; Freire, Thamires Batello; Pinto, Claudinéia Aparecida Sales de Oliveira; Prado, María Segunda Aurora; Baby, André R; Velasco, Maria Valéria R

    2017-10-15

    Tryptophan, an amino acid found in hair proteinaceous structure is used as a marker of hair photodegradation. Also, protein loss caused by several chemical/physical treatments can be inferred by tryptophan quantification. Kynurenine is a photo-oxidation product of tryptophan, expected to be detected when hair is exposed mainly to UVB (290-320nm) radiation range. Tryptophan from hair is usually quantified directly as a solid or after alkaline hydrolysis, spectrofluorimetrically. However, these types of measure are not sufficiently specific and present several interfering substances. Thus, this work aimed to propose a quantification method for both tryptophan and kynurenine in hair samples, after alkali hydrolysis process, by using high-performance liquid chromatography (HPLC) with fluorimetric and UV detection. The tryptophan and kynurenine quantification method was developed and validated. Black, white, bleached and dyed (blond and auburn) hair tresses were used in this study. Tryptophan and kynurenine were separated within ∼9min by HPLC. Both black and white virgin hair samples presented similar concentrations of tryptophan, while bleaching caused a reduction in the tryptophan content as well as dyeing process. Unexpectedly, UV/vis radiation did not promote significantly the conversion of tryptophan into its photo-oxidation product and consequently, kynurenine was not detected. Thus, this works presented an acceptable method for quantification of tryptophan and its photooxidation metabolite kynurenine in hair samples. Also, the results indicated that bleaching and dyeing processes promoted protein/amino acids loss but tryptophan is not extensively degraded in human hair by solar radiation. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Dynamic Determinants of the Uncontrolled Manifold during Human Quiet Stance.

    Science.gov (United States)

    Suzuki, Yasuyuki; Morimoto, Hiroki; Kiyono, Ken; Morasso, Pietro G; Nomura, Taishin

    2016-01-01

    Human postural sway during stance arises from coordinated multi-joint movements. Thus, a sway trajectory represented by a time-varying postural vector in the multiple-joint-angle-space tends to be constrained to a low-dimensional subspace. It has been proposed that the subspace corresponds to a manifold defined by a kinematic constraint, such that the position of the center of mass (CoM) of the whole body is constant in time, referred to as the kinematic uncontrolled manifold ( kinematic-UCM ). A control strategy related to this hypothesis ( CoM-control-strategy ) claims that the central nervous system (CNS) aims to keep the posture close to the kinematic-UCM using a continuous feedback controller, leading to sway patterns that mostly occur within the kinematic-UCM, where no corrective control is exerted. An alternative strategy proposed by the authors ( intermittent control-strategy ) claims that the CNS stabilizes posture by intermittently suspending the active feedback controller, in such a way to allow the CNS to exploit a stable manifold of the saddle-type upright equilibrium in the state-space of the system, referred to as the dynamic-UCM , when the state point is on or near the manifold. Although the mathematical definitions of the kinematic- and dynamic-UCM are completely different, both UCMs play similar roles in the stabilization of multi-joint upright posture. The purpose of this study was to compare the dynamic performance of the two control strategies. In particular, we considered a double-inverted-pendulum-model of postural control, and analyzed the two UCMs defined above. We first showed that the geometric configurations of the two UCMs are almost identical. We then investigated whether the UCM-component of experimental sway could be considered as passive dynamics with no active control, and showed that such UCM-component mainly consists of high frequency oscillations above 1 Hz, corresponding to anti-phase coordination between the ankle and hip. We

  8. Dynamic determinants of the uncontrolled manifold during human quiet stance

    Directory of Open Access Journals (Sweden)

    Yasuyuki Suzuki

    2016-12-01

    Full Text Available Human postural sway during stance arises from coordinated multi-joint movements. Thus, a sway trajectory represented by a time-varying postural vector in the multiple-joint-angle-space tends to be constrained to a low-dimensional subspace. It has been proposed that the subspace corresponds to a manifold defined by a kinematic constraint, such that the position of the center of mass (CoM of the whole body is constant in time, referred to as the kinematic uncontrolled manifold (kinematic-UCM. A control strategy related to this hypothesis (CoM-control-strategy claims that the central nervous system (CNS aims to keep the posture close to the kinematic-UCM using a continuous feedback controller, leading to sway patterns that mostly occur within the kinematic-UCM, where no corrective control is exerted. An alternative strategy proposed by the authors (intermittent control-strategy claims that the CNS stabilizes posture by intermittently suspending the active feedback controller, in such a way to allow the CNS to exploit a stable manifold of the saddle-type upright equilibrium in the state-space of the system, referred to as the dynamic-UCM, when the state point is on or near the manifold. Although the mathematical definitions of the kinematic- and dynamic-UCM are completely different, both UCMs play similar roles in the stabilization of multi-joint upright posture. The purpose of this study was to compare the dynamic performance of the two control strategies. In particular, we considered a double-inverted-pendulum-model of postural control, and analyzed the two UCMs defined above. We first showed that the geometric configurations of the two UCMs are almost identical. We then investigated whether the UCM-component of experimental sway could be considered as passive dynamics with no active control, and showed that such UCM-component mainly consists of high frequency oscillations above 1 Hz, corresponding to anti-phase coordination between the ankle and

  9. Standardization for cortisol determination in human blood by competitive protein-binding

    International Nuclear Information System (INIS)

    Okada, H.

    1978-01-01

    Standardization for determination of cortisol from human plasma (17-hydroxycorticosteroids) using competitive protein-binding method is presented. Activated carbon coated with dextrans is used for separation of the hormone-protein complexe and hormone labelled free [pt

  10. 77 FR 38374 - Culturally Significant Objects Imported for Exhibition Determinations: “The Human Beast: German...

    Science.gov (United States)

    2012-06-27

    ... DEPARTMENT OF STATE [Public Notice 7935] Culturally Significant Objects Imported for Exhibition Determinations: ``The Human Beast: German Expressionism at The San Diego Museum of Art'' SUMMARY: Notice is... objects to be included in the exhibition ``The Human Beast: German Expressionism at The San Diego Museum...

  11. Determination of platinum in human subcellular microsamples by inductively coupled plasma mass spectrometry

    DEFF Research Database (Denmark)

    Björn, Erik; Nygren, Yvonne; Nguyen, Tam T. T. N.

    2007-01-01

    A fast and robust method for the determination of platinum in human subcellular microsamples by inductively coupled plasma mass spectrometry was developed, characterized, and validated. Samples of isolated DNA and exosome fractions from human ovarian (2008) and melanoma (T289) cancer cell lines w...

  12. Major differences between human atopic dermatitis and murine models as determined by global transcriptomic profiling

    DEFF Research Database (Denmark)

    Ewald, David Adrian; Noda, Shinji; Oliva, Margeaux

    2017-01-01

    , and a comparison of these models with the human AD transcriptomic fingerprint is lacking. We sought to evaluate the transcriptomic profiles of six common murine models and determine how they relate to human AD skin. Transcriptomic profiling was performed using microarrays and qRT-PCR on biopsies from NC/Nga, flaky...

  13. Determining adaptive and adverse oxidative stress responses in human bronical epithelial cells exposed to zinc

    Science.gov (United States)

    Determining adaptive and adverse oxidative stress responses in human bronchial epithelial cells exposed to zincJenna M. Currier1,2, Wan-Yun Cheng1, Rory Conolly1, Brian N. Chorley1Zinc is a ubiquitous contaminant of ambient air that presents an oxidant challenge to the human lung...

  14. Determining Human Clot Lysis Time (in vitro with Plasminogen/Plasmin from Four Species (Human, Bovine, Goat, and Swine

    Directory of Open Access Journals (Sweden)

    Omaira Cañas Bermúdez

    2015-05-01

    Full Text Available Cardiovascular disease is the leading cause of death worldwide, including failures in the plasminogen/plasmin system which is an important factor in poor lysis of blood clots. This article studies the fibrinolytic system in four species of mammals, and it identifies human plasminogen with highest thrombolysis efficiency. It examines plasminogen from four species (human, bovine, goat, and swine and identifies the most efficient one in human clot lysis in vitro. All plasminogens were identically purified by affinity chromatography. Human fibrinogen was purified by fractionation with ethanol. The purification of both plasminogen and fibrinogen was characterized by one-dimensional SDS-PAGE (10%. Human clot formation in vitro and its dissolution by plasminogen/plasmin consisted of determining lysis time from clot formation to its dilution. Purification of proteins showed greater than 95% purity, human plasminogen showed greater ability to lyse clot than animal plasminogen. The article concludes that human plasminogen/plasmin has the greatest catalysis and efficiency, as it dissolves human clot up to three times faster than that of irrational species.

  15. Effects of human errors on the determination of surveillance test interval

    International Nuclear Information System (INIS)

    Chung, Dae Wook; Koo, Bon Hyun

    1990-01-01

    This paper incorporates the effects of human error relevant to the periodic test on the unavailability of the safety system as well as the component unavailability. Two types of possible human error during the test are considered. One is the possibility that a good safety system is inadvertently left in a bad state after the test (Type A human error) and the other is the possibility that bad safety system is undetected upon the test (Type B human error). An event tree model is developed for the steady-state unavailability of safety system to determine the effects of human errors on the component unavailability and the test interval. We perform the reliability analysis of safety injection system (SIS) by applying aforementioned two types of human error to safety injection pumps. Results of various sensitivity analyses show that; 1) the appropriate test interval decreases and steady-state unavailability increases as the probabilities of both types of human errors increase, and they are far more sensitive to Type A human error than Type B and 2) the SIS unavailability increases slightly as the probability of Type B human error increases, and significantly as the probability of Type A human error increases. Therefore, to avoid underestimation, the effects of human error should be incorporated in the system reliability analysis which aims at the relaxations of the surveillance test intervals, and Type A human error has more important effect on the unavailability and surveillance test interval

  16. Using indicators to determine the contribution of human rights to public health efforts

    Science.gov (United States)

    Ferguson, Laura

    2009-01-01

    Abstract There is general agreement on the need to integrate human rights into health policies and programmes, although there is still reluctance to go beyond rhetorical acknowledgement of their assumed significance. To determine the actual value of human rights for the effectiveness of public health efforts requires clarity about what their incorporation looks like in practice and how to assess their contribution. Despite the pervasive use of indicators in the public health field, indicators that specifically capture human rights concerns are not well developed and those that exist are inconsistently used. Even though “health and human rights indicators” are increasingly being constructed, it is often the case that health indicators are used to draw conclusions about some interaction between human rights and health; or that law and policy or other indicators, traditionally the domain of the human rights community, are used to make conclusions about health outcomes. To capture the added value that human rights bring to health, the differences in the contributions offered by these indicators need to be understood. To determine the value of different measures for advancing programme effectiveness, improving health outcomes and promoting human rights, requires questioning the intended purpose behind the construction of an indicator, who uses it, the kind of indicator it is, the extent to which it provides information about vulnerable populations, as well as how the data are collected and used. PMID:19784452

  17. Higher cortical modulation of pain perception in the human brain: Psychological determinant.

    Science.gov (United States)

    Chen, Andrew Cn

    2009-10-01

    Pain perception and its genesis in the human brain have been reviewed recently. In the current article, the reports on pain modulation in the human brain were reviewed from higher cortical regulation, i.e. top-down effect, particularly studied in psychological determinants. Pain modulation can be examined by gene therapy, physical modulation, pharmacological modulation, psychological modulation, and pathophysiological modulation. In psychological modulation, this article examined (a) willed determination, (b) distraction, (c) placebo, (d) hypnosis, (e) meditation, (f) qi-gong, (g) belief, and (h) emotions, respectively, in the brain function for pain modulation. In each, the operational definition, cortical processing, neuroimaging, and pain modulation were systematically deliberated. However, not all studies had featured the brain modulation processing but rather demonstrated potential effects on human pain. In our own studies on the emotional modulation on human pain, we observed that emotions could be induced from music melodies or pictures perception for reduction of tonic human pain, mainly in potentiation of the posterior alpha EEG fields, likely resulted from underneath activities of precuneous in regulation of consciousness, including pain perception. To sum, higher brain functions become the leading edge research in all sciences. How to solve the information bit of thinking and feeling in the brain can be the greatest challenge of human intelligence. Application of higher cortical modulation of human pain and suffering can lead to the progress of social humanity and civilization.

  18. Higher cortical modulation of pain perception in the human brain: Psychological determinant

    OpenAIRE

    Chen, Andrew Cn

    2009-01-01

    Pain perception and its genesis in the human brain have been reviewed recently. In the current article, the reports on pain modulation in the human brain were reviewed from higher cortical regulation, i.e. top-down effect, particularly studied in psychological determinants. Pain modulation can be examined by gene therapy, physical modulation, pharmacological modulation, psychological modulation, and pathophysiological modulation. In psychological modulation, this article examined (a) willed d...

  19. The 5-HT2A receptor binding pattern in the human brain is strongly genetically determined

    DEFF Research Database (Denmark)

    Pinborg, Lars H; Arfan, Haroon; Haugbol, Steven

    2007-01-01

    With the appropriate radiolabeled tracers, positron emission tomography (PET) enables in vivo human brain imaging of markers for neurotransmission, including neurotransmitter synthesis, receptors, and transporters. Whereas structural imaging studies have provided compelling evidence that the human...... brain anatomy is largely genetically determined, it is currently unknown to what degree neuromodulatory markers are subjected to genetic and environmental influence. Changes in serotonin 2A (5-HT(2A)) receptors have been reported to occur in various neuropsychiatric disorders and an association between...

  20. SOCIOECONOMIC, INSTITUTIONAL & POLITICAL DETERMINANTS OF HUMAN RIGHTS ABUSES: A SUBNATIONAL STUDY OF INDIA, 1993 – 2002

    OpenAIRE

    Vadlamannati, Krishna Chaitanya

    2008-01-01

    We conduct an econometric analysis of socioeconomic, institutional and political factors determining government respect for human rights within India. Using time series cross-sectional data for 28 Indian states for the period 1993 – 2002, we find that internal threat poised by number of social violence events, presence of civil war and riot hit disturbed areas are strongly associated with human rights abuses. Amongst socioeconomic factors, ‘exclusive’ economic growth, ‘uneven’ development, po...

  1. Predictive values of traditional animal bioassay studies for human perinatal carcinogenesis risk determination

    International Nuclear Information System (INIS)

    Anderson, Lucy M.

    2004-01-01

    The many physiological, biochemical, and structure differences between rodents and humans, especially with regard to gestation and fetal development, invite questions as to the utility of rodent models for the prediction of risk of perinatal carcinogenesis in humans and for extrapolation of mechanistic studies. Here, the relevance of basic generalities, derived from rodent perinatal studies, to human contexts is considered. The cross-species usefulness of these generalities was upheld by the example of carcinogen activation and detoxification as determining factors. These have been established in rodent studies and recently indicted in humans by investigations of genetic polymorphisms in cytochromes P450, N-acetyltransferase, myeloperoxidase, quinone reductase, and glutathione S-transferase. Also, published data have been analyzed comparatively for diethylstilbestrol and irradiation, the two known human transplacental carcinogenic agents. At similar doses to those experienced by humans, both diethylstilbestrol and X- and gamma-irradiation in rodents and dogs yielded increased tumors at rates similar to those for humans. In rodents, there was a clearly negative relationship between total diethylstilbestrol dose and tumors per dose unit, and a similar pattern was suggested for radiation. Diethylstilbestrol had transgenerational effects that did not diminish over three generations. Overall, this analysis of the published literature indicates that there are basic qualitative and quantitative similarities in the responsiveness of human and rodent fetuses to carcinogens, and that dose effects may be complex and in need of further investigation

  2. Human skeletal muscle drug transporters determine local exposure and toxicity of statins.

    Science.gov (United States)

    Knauer, Michael J; Urquhart, Bradley L; Meyer zu Schwabedissen, Henriette E; Schwarz, Ute I; Lemke, Christopher J; Leake, Brenda F; Kim, Richard B; Tirona, Rommel G

    2010-02-05

    The 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, or statins, are important drugs used in the treatment and prevention of cardiovascular disease. Although statins are well tolerated, many patients develop myopathy manifesting as muscle aches and pain. Rhabdomyolysis is a rare but severe toxicity of statins. Interindividual differences in the activities of hepatic membrane drug transporters and metabolic enzymes are known to influence statin plasma pharmacokinetics and risk for myopathy. Interestingly, little is known regarding the molecular determinants of statin distribution into skeletal muscle and its relevance to toxicity. We sought to identify statin transporters in human skeletal muscle and determine their impact on statin toxicity in vitro. We demonstrate that the uptake transporter OATP2B1 (human organic anion transporting polypeptide 2B1) and the efflux transporters, multidrug resistance-associated protein (MRP)1, MRP4, and MRP5 are expressed on the sarcolemmal membrane of human skeletal muscle fibers and that atorvastatin and rosuvastatin are substrates of these transporters when assessed using a heterologous expression system. In an in vitro model of differentiated, primary human skeletal muscle myoblast cells, we demonstrate basal membrane expression and drug efflux activity of MRP1, which contributes to reducing intracellular statin accumulation. Furthermore, we show that expression of human OATP2B1 in human skeletal muscle myoblast cells by adenoviral vectors increases intracellular accumulation and toxicity of statins and such effects were abrogated when cells overexpressed MRP1. These results identify key membrane transporters as modulators of skeletal muscle statin exposure and toxicity.

  3. Determination of ribavirin in human serum using liquid chromatography tandem mass spectrometry

    NARCIS (Netherlands)

    van der Lijke, H.; Alffenaar, J.-W. C.; Kok, W.Th.; Greijdanus, B.; Uges, D.R.A.

    2012-01-01

    A method has been developed for the determination of ribavirin in human serum for therapeutic drug monitoring purposes, using liquid chromatography electrospray ionization mass spectrometry. Separation was obtained with a mobile phase gradient starting and ending in 100% aqueous conditions using a

  4. A Laboratory Exercise to Determine Human ABO Blood Type by Noninvasive Methods

    Science.gov (United States)

    Martin, Michael P.; Detzel, Stephen M.

    2008-01-01

    Analysis of single-nucleotide polymorphisms and their association with diseases and nondisease phenotypes is of growing importance in human biology studies. In this laboratory exercise, students determine the genetic basis for their ABO blood type; however, no blood is drawn. Students isolate genomic DNA from buccal mucosa cells that are present…

  5. Determinants of Human Papillomavirus Vaccination Intention Among Female Sex Workers in Amsterdam, the Netherlands

    NARCIS (Netherlands)

    Marra, E.; Dam, L. van; Kroone, N.; Alberts, C.J.; Craanen, M.; Zimet, G.D.; Heijmam, T.; Hogewoning, A.A.; Sonder, G.J.B.; Vries, H.J.C. de; Alberts, C.J.; Paulussen, T.G.W.M.; Schim van der Loeff, M.F.

    2017-01-01

    Introduction: Female sex workers (FSWs) are at risk for human papillomavirus (HPV)-induced diseases but are currently not targeted by the HPV vaccination program in the Netherlands. We explored determinants of their intention to get vaccinated against HPV in case vaccination would be offered to

  6. Palmitate and stearate binding to human serum albumin. Determination of relative binding constants

    DEFF Research Database (Denmark)

    Vorum, H; Fisker, K; Honoré, B

    1997-01-01

    Multiple binding equilibria of two apparently insoluble ligands, palmitate and stearate, to defatted human serum albumin were studied in a 66 mM sodium phosphate buffer (pH 7.4) at 37 degrees C, by determination of dialytic exchange rates of ligands among identical equilibrium solutions. The expe...

  7. Teachers' Conceptions About the Genetic Determinism of Human Behaviour: A Survey in 23 Countries

    Science.gov (United States)

    Castéra, Jérémy; Clément, Pierre

    2014-02-01

    This work analyses the answers to a questionnaire from 8,285 in-service and pre-service teachers from 23 countries, elaborated by the Biohead-Citizen research project, to investigate teachers' conceptions related to the genetic determinism of human behaviour. A principal components analysis is used to assess the main trends in all the interviewed teachers' conceptions. This illustrates that innatism is present in two distinct ways: in relation to individuals (e.g. genetic determinism to justify intellectual likeness between individuals such as twins) or in relation to groups of humans (e.g. genetic determinism to justify gender differences or the superiority of some human ethnic groups). A between-factor analysis discriminates between countries, showing very significant differences. There is more innatism among teachers' conceptions in African countries and Lebanon than in European countries, Brazil and Australia. Among the other controlled parameters, only two are significantly independent of the country: the level of training and the level of knowledge of biology. A co-inertia analysis shows a strong correlation between non-citizen attitudes towards and innatist conceptions of genetic determinism regarding human groups. We discuss these findings and their implications for education.

  8. Determination of Elements in Normal and Leukemic Human Whole Blood by Neutron Activation Analysis

    Energy Technology Data Exchange (ETDEWEB)

    Brune, D; Frykberg, B; Samsahl, K; Wester, P O

    1961-11-15

    By means of gamma-spectrometry the following elements were simultaneously determined in normal and leukemic human whole blood: Cu, Mn, Zn, Sr, Na, P, Ca, Rb, Cd, Sb, Au, Cs and Fe. Chemical separations were performed according to a group separation method using ion-exchange technique. No significant difference between the concentrations of the elements in normal- and leukemic blood was observed.

  9. LC determination of propylene glycol in human plasma after pre-column derivatization with benzoyl chloride

    NARCIS (Netherlands)

    Sinjewel, A.; Swart, E.L.; Lingeman, H.; Wilhelm, A.J.

    2007-01-01

    A simple high-performance liquid chromatographic method, using photodiode array detection was developed for the determination of propylene glycol in human plasma and in the fluid retreived after continuous veno-venous hemofiltration. The method entailed alkaline derivatization with benzoyl chloride

  10. Susceptibility of human and probiotic Bifidobacterium spp. to selected antibiotics as determined by the Etest method

    NARCIS (Netherlands)

    Matto, J.; Hoek, van A.H.A.M.; Domig, K.J.; Saarela, M.; Flórez, A.B.; Brockmann, E.; Amtmann, E.; Mayo, B.; Aarts, H.J.M.; Danielsen, M.

    2007-01-01

    This study reports the antibiotic susceptibility of 203 strains representing human or probiotic associated Bifidobacterium species as determined by the Etest method. Strains showing minimum inhibitory concentration (MIC) for tetracycline >= 16 mu g mL(-1) were detected in all studied

  11. Determination of telmisartan in human blood plasma: Part I: Immunoassay development

    NARCIS (Netherlands)

    Hempen, C.M.; Gläsle-Schwarz, Liane; Kunz, Ulrich; Karst, U.

    2006-01-01

    Telmisartan is an angiotensin II receptor antagonist and a known drug against high blood pressure. In this report, the development of a new and rapid analytical technique, an enzyme-linked immunosorbent assay (ELISA) for the determination of telmisartan in human blood plasma is described. The

  12. Teachers' Conceptions about the Genetic Determinism of Human Behaviour: A Survey in 23 Countries

    Science.gov (United States)

    Castéra, Jérémy; Clément, Pierre

    2014-01-01

    This work analyses the answers to a questionnaire from 8,285 in-service and pre-service teachers from 23 countries, elaborated by the Biohead-Citizen research project, to investigate teachers' conceptions related to the genetic determinism of human behaviour. A principal components analysis is used to assess the main trends in all the interviewed…

  13. Cultural, Human, and Social Capital as Determinants of Corporal Punishment: Toward an Integrated Theoretical Model.

    Science.gov (United States)

    Xu, Xiaohe; Tung, Yuk-Ying; Dunaway, R. Gregory

    2000-01-01

    This article constructs a model to predict the likelihood of parental use of corporal punishment on children in two-parent families. Reports that corporal punishment is primarily determined by cultural, human, and social capital that are available to, or already acquired by parents. Discusses an integrated, resource-based theory for predicting use…

  14. Determination of Elements in Normal and Leukemic Human Whole Blood by Neutron Activation Analysis

    International Nuclear Information System (INIS)

    Brune, D.; Frykberg, B.; Samsahl, K.; Wester, P.O.

    1961-11-01

    By means of gamma-spectrometry the following elements were simultaneously determined in normal and leukemic human whole blood: Cu, Mn, Zn, Sr, Na, P, Ca, Rb, Cd, Sb, Au, Cs and Fe. Chemical separations were performed according to a group separation method using ion-exchange technique. No significant difference between the concentrations of the elements in normal- and leukemic blood was observed

  15. Human Capital, Social Classes, and the Earnings Determination Process in Brazilian Agriculture.

    Science.gov (United States)

    Neves, Jorge A.; Haller, Archibald O.; Fernandes, Danielle C.

    This paper examines the process of earnings determination in the agricultural sector of Brazil. Among the main causal factors analyzed are human capital (education and work experience), labor market segmentation, gender, social class position, level of development/modernization, and concentration of land ownership. Data on individuals employed in…

  16. Training-induced changes in peritendinous type I collagen turnover determined by microdialysis in humans

    DEFF Research Database (Denmark)

    Langberg, Henning; Rosendal, L; Kjaer, M

    2001-01-01

    1. Acute exercise is found to increase collagen type I formation locally in peritendinous connective tissue of the Achilles' tendon in humans, as determined from changes in interstitial concentrations of collagen propeptide (PICP) and a collagen degradation product (ICTP) by the use of microdialy...

  17. Online immunocapture ICP-MS for the determination of the metalloprotein ceruloplasmin in human serum.

    Science.gov (United States)

    Bernevic, Bogdan; El-Khatib, Ahmed H; Jakubowski, Norbert; Weller, Michael G

    2018-04-02

    The human copper-protein ceruloplasmin (Cp) is the major copper-containing protein in the human body. The accurate determination of Cp is mandatory for the reliable diagnosis of several diseases. However, the analysis of Cp has proven to be difficult. The aim of our work was a proof of concept for the determination of a metalloprotein-based on online immunocapture ICP-MS. The immuno-affinity step is responsible for the enrichment and isolation of the analyte from serum, whereas the compound-independent quantitation with ICP-MS delivers the sensitivity, precision, and large dynamic range. Off-line ELISA (enzyme-linked immunosorbent assay) was used in parallel to confirm the elution profile of the analyte with a structure-selective method. The total protein elution was observed with the 32 S mass trace. The ICP-MS signals were normalized on a 59 Co signal. The human copper-protein Cp could be selectively determined. This was shown with pure Cp and with a sample of human serum. The good correlation with off-line ELISA shows that Cp could be captured and eluted selectively from the anti-Cp affinity column and subsequently determined by the copper signal of ICP-MS.

  18. [Neurobiological determinism: questionable inferences on human freedom of choice and forensic criminal responsibility].

    Science.gov (United States)

    Urbaniok, F; Hardegger, J; Rossegger, A; Endrass, J

    2006-08-01

    Several authors argue that criminal behavior is generally caused by neurobiological deficits. Based on this neurobiological perspective of assumed causality, the concept of free will is questioned, and the theory of neurobiological determinism of all human behavior is put forward, thus maintaining that human beings are not responsible for their actions, and consequently the principle of guilt should be given up in criminal law. In this context the controversial debate on determinism and indeterminism, which has been held for centuries, has flared up anew, especially within the science of criminal law. When critically examining the current state of research, it becomes apparent that the results do not support the existence of a universally valid neurobiological causality of criminal behavior, nor a theory of an absolute neurobiological determinism. Neither is complete determination of all phenomena in the universe--as maintained--the logical conclusion of the principle of causality, nor is it empirically confirmed. Analyzed methodically, it cannot be falsified, and thus, as a theory which cannot be empirically tested, it represents a dogma against which plausible objections can be made. The criticism of the concept of free will, and even more so of human accountability and criminal responsibility, is not put forward in a valid way. The principle of relative determinism--the evaluation of the degree of determinism of personality factors potentially reducing criminal responsibility, which includes concrete observations and analysis of behavior--thus remains a central and cogent approach to the assessment of criminal responsibility. To sum up, the theories proposed by some authors on the complete neurobiological determinism of human behavior, and the subsequent impossibility of individual responsibility and guilt, reveal both methodical misconception and a lack of empirical foundation.

  19. ALFY-Controlled DVL3 Autophagy Regulates Wnt Signaling, Determining Human Brain Size.

    Directory of Open Access Journals (Sweden)

    Rotem Kadir

    2016-03-01

    Full Text Available Primary microcephaly is a congenital neurodevelopmental disorder of reduced head circumference and brain volume, with fewer neurons in the cortex of the developing brain due to premature transition between symmetrical and asymmetrical cellular division of the neuronal stem cell layer during neurogenesis. We now show through linkage analysis and whole exome sequencing, that a dominant mutation in ALFY, encoding an autophagy scaffold protein, causes human primary microcephaly. We demonstrate the dominant effect of the mutation in drosophila: transgenic flies harboring the human mutant allele display small brain volume, recapitulating the disease phenotype. Moreover, eye-specific expression of human mutant ALFY causes rough eye phenotype. In molecular terms, we demonstrate that normally ALFY attenuates the canonical Wnt signaling pathway via autophagy-dependent removal specifically of aggregates of DVL3 and not of Dvl1 or Dvl2. Thus, autophagic attenuation of Wnt signaling through removal of Dvl3 aggregates by ALFY acts in determining human brain size.

  20. Non-alcoholic fatty liver disease and impaired proinsulin conversion as newly identified predictors of the long-term non-response to a lifestyle intervention for diabetes prevention: results from the TULIP study.

    Science.gov (United States)

    Schmid, Vera; Wagner, Robert; Sailer, Corinna; Fritsche, Louise; Kantartzis, Konstantinos; Peter, Andreas; Heni, Martin; Häring, Hans-Ulrich; Stefan, Norbert; Fritsche, Andreas

    2017-12-01

    Lifestyle intervention is effective to prevent type 2 diabetes. However, a considerable long-term non-response occurs to a standard lifestyle intervention. We investigated which risk phenotypes at baseline and their changes during the lifestyle intervention predict long-term glycaemic non-response to the intervention. Of 300 participants at high risk for type 2 diabetes who participated in a 24 month lifestyle intervention with diet modification and increased physical activity, 190 participants could be re-examined after 8.7 ± 1.6 years. All individuals underwent a five-point 75 g OGTT and measurements of body fat compartments and liver fat content with MRI and spectroscopy at baseline, 9 and 24 months during the lifestyle intervention, and at long-term follow-up. Fasting proinsulin to insulin conversion (PI/I ratio) and insulin sensitivity and secretion were calculated from the OGTT. Non-response to lifestyle intervention was defined as no decrease in glycaemia, i.e. no decrease in AUC for glucose at 0-120 min during OGTT (AUCglucose 0-120 min ). Before the lifestyle intervention, 56% of participants had normal glucose regulation and 44% individuals had impaired fasting glucose and/or impaired glucose tolerance. At long-term follow-up, 11% had developed diabetes. Multivariable regression analysis with adjustment for age, sex, BMI and change in BMI during the lifestyle intervention revealed that baseline insulin secretion and insulin sensitivity, as well as change in insulin sensitivity during the lifestyle intervention, predicted long-term glycaemic control after 9 years. In addition, increased hepatic lipid content as well as impaired fasting proinsulin conversion at baseline were newly detected phenotypes that independently predicted long-term glycaemic control. Increased hepatic lipid content and impaired proinsulin conversion are new predictors, independent of change in body weight, for non-response to lifestyle intervention in addition to the

  1. Multi-element determination in cancellous bone of human femoral head by PIXE

    International Nuclear Information System (INIS)

    Yuanxun Zhang; Yongping Zhang; Yongpeng Tong; Shijing Qiu; Xiaotao Wu; Kerong Dai

    1996-01-01

    Proton Induced X-ray Emission (PIXE) method is used for the determination of elemental concentrations in cancellous bone of human femoral head from five autopsies and seven patients with femoral neck broken. The specimen preparation and experimental procedure are described in detail. Using the t test, the results show that the concentrations of P, Ca, Fe, Cu, Sr in control group are higher than those in patient group, but the concentrations of S, K, Zn, Mn are not significantly different. The physiological functions of metallic elements in human bone are also discussed. (author). 19 refs., 1 fig., 4 tabs

  2. Determination of cytotoxicity in vivo using 111Indium-labelled human tumor cells

    International Nuclear Information System (INIS)

    Lockshin, Arnold; Giovanella, B.C.; Kolielski, Tony; Stehlin, J.S. Jr.

    1984-01-01

    Loss of radioactivity from nude mice was determined after inoculation of human tumor cells prelabelled with ( 111 In)indium oxine ( 111 InOx). Elimination of 111 In was increased somewhat by treating the mice with diphtheria toxin (DT), which is toxic selectively for human cells compared to mice. Calcium disodium edetate (CaNa 2 EDTA), a metal chelating agent, facilitated elimination of 111 In and increased the difference in the rates of loss of radioactivity from mice bearing viable compared to DT-killed cells. (author)

  3. Validation method for determination of cholesterol in human urine with electrochemical sensors using gold electrodes

    Science.gov (United States)

    Riyanto, Laksono, Tomy Agung

    2017-12-01

    Electrochemical sensors for the determination of cholesterol with Au as a working electrode (Au) and its application to the analysis of urine have been done. The gold electrode was prepared using gold pure (99.99%), with size 1.0 mm by length and wide respectively, connected with silver wire using silver conductive paint. Validation methods have been investigated in the analysis of cholesterol in human urine using electrochemical sensors or cyclic voltammetry (CV) method. The effect of electrolyte and uric acid concentration has been determined to produce the optimum method. Validation method parameters for cholesterol analysis in human urine using CV are precision, recovery, linearity, limit of detection (LOD) and limit of quantification (LOQ). The result showed the correlation of concentration of cholesterol to anodic peak current is the coefficient determination of R2 = 0.916. The results of the validation method showed the precision, recovery, linearity, LOD, and LOQ are 1.2539%, 144.33%, 0.916, 1.49 × 10-1 mM and 4.96 × 10-1 mM, respectively. As a conclusion is Au electrode is a good electrode for electrochemical sensors to determination of cholesterol in human urine.

  4. Gut luminal endogenous protein: implications for the determination of ileal amino acid digestibility in humans.

    Science.gov (United States)

    Moughan, Paul J; Rutherfurd, Shane M

    2012-08-01

    The true ileal digestibility assay provides the most informative measure of digestibility to assess bioavailability of amino acids in foods for humans. To determine 'true' estimates of ileal amino acid digestibility, requires that endogenous amino acids present in digesta at the terminal ileum be quantified. The amounts of endogenous amino acids in ileal digesta can be determined after feeding an animal or human a protein-free diet (traditional approach) or by various methods after giving a protein-containing diet. When the protein-free method has been applied with adult human subjects an overall mean value (three separate studies) for endogenous ileal nitrogen flow of 800 mg N/d has been reported. This value is considerably lower than a comparable value obtained after feeding protein of 1852 mg N/d (mean of four separate studies), and thus endogenous ileal N and amino acids should be measured under conditions of protein alimentation. There is some confusion concerning the terminology used to define digestibility, with the term "true" digestibility having different adopted meanings. Here, true amino acid digestibility is defined as apparent amino acid digestibility corrected for the basal amino acid losses determined after giving either a protein-free or a protein-containing diet. Basal losses should be determined at a defined dry-matter and protein intake. The protein-free diet approach to determining endogenous amino acids is considered unphysiological and basal losses refer to ileal endogenous amino acid flows associated with digesta dry-matter flow, and not including "specific" effects of dietary factors such as non starch polysaccharides and anti nutritional factors. Arguments are advanced that the enzyme hydrolysed protein/ultra filtration method may be suitable for routine application with a cannulated pig model, to obtain physiologically-valid basal estimates of ileal endogenous amino acids to allow calculation of true ileal amino acid digestibility in the

  5. Biological psychological and social determinants of old age: Bio-psycho-social aspects of human aging

    Directory of Open Access Journals (Sweden)

    Małgorzata Dziechciaż

    2014-11-01

    Full Text Available Biological psychological and social determinants of old age: Bio-psycho-social aspects of human aging. The aging of humans is a physiological and dynamic process ongoing with time. In accordance with most gerontologists’ assertions it starts in the fourth decade of life and leads to death. The process of human aging is complex and individualized, occurs in the biological, psychological and social sphere. Biological aging is characterized by progressive age-changes in metabolism and physicochemical properties of cells, leading to impaired self-regulation, regeneration, and to structural changes and functional tissues and organs. It is a natural and irreversible process which can run as successful aging, typical or pathological. Biological changes that occur with age in the human body affect mood, attitude to the environment, physical condition and social activity, and designate the place of seniors in the family and society. Psychical ageing refers to human awareness and his adaptability to the ageing process. Among adaptation attitudes we can differentiate: constructive, dependence, hostile towards others and towards self attitudes. With progressed age, difficulties with adjustment to the new situation are increasing, adverse changes in the cognitive and intellectual sphere take place, perception process involutes, perceived sensations and information received is lowered, and thinking processes change. Social ageing is limited to the role of an old person is culturally conditioned and may change as customs change. Social ageing refers to how a human being perceives the ageing process and how society sees it.

  6. Determination of fluvoxamine maleate in human urine and human serum using alkaline KMnO4 -rhodamine B chemiluminescence.

    Science.gov (United States)

    Yang, Dongqin; He, Yanyan; Chen, Funan

    2017-09-01

    The flow-injection chemiluminescence (FI-CL) behavior of a gold nanocluster (Au NC)-enhanced rhodamine B-KMnO 4 system was studied under alkaline conditions for the first time. In the present study, the as-prepared bovine serum albumin-stabilized Au NCs showed excellent stability and reproducibility. The addition of trace levels of fluvoxamine maleate (Flu) led to an obvious decline in CL intensity in the rhodamine B-KMnO 4 -Au NCs system, which could be used for quantitative detection of Flu. Under optimized conditions, the proposed CL system exhibited a favorable analytical performance for Flu determination in the range 2 to 100 μg ml -1 . The detection limit for Flu measurement was 0.021 μg ml -1 . Moreover, this newly developed system revealed outstanding selectivity for Flu detection when compared with a multitude of other species, such as the usual ions, uric acid and a section of hydroxy compounds. Additionally, CL spectra, UV-visible spectroscopes and fluorescence spectra were measured in order to determine the possible reaction mechanism. This approach could be used to detect Flu in human urine and human serum samples with the desired recoveries and could have promising application under physiological conditions. Copyright © 2017 John Wiley & Sons, Ltd.

  7. Concentration of 24 Trace Elements in Human Heart Tissue Determined by Neutron Activation Analysis

    Energy Technology Data Exchange (ETDEWEB)

    Wester, P O

    1964-06-15

    By means of neutron-activation analysis, human heart tissue from autopsy of 20 victims of traumatic accidents has been investigated with respect to the concentration of 24 different trace elements. A recently developed ion-exchange technique combined with gamma spectrometry has been used, which permits simultaneous determination of a large number of trace elements. The following trace elements have been determined quantitatively: Ag, As, Au, Ba, Br; Ca, Cd, Ce, Co, Cr, Cs, Cu, Fe, Hg, La, Mo, Pt, Rb, Sb, Se, Se, Sm, Zn, W. In some heart samples, Hf and Os were determined qualitatively. The mean and standard deviation are given for the elements Cu, Fe, Se and Zn, Since none of the other quantitatively determined trace elements were normally distributed, the median is given as the central value. When possible, comparisons with values from other investigations have been made. No marked differences in the trace-element concentrations with age or sex could be detected.

  8. Concentration of 24 Trace Elements in Human Heart Tissue Determined by Neutron Activation Analysis

    International Nuclear Information System (INIS)

    Wester, P.O.

    1964-06-01

    By means of neutron-activation analysis, human heart tissue from autopsy of 20 victims of traumatic accidents has been investigated with respect to the concentration of 24 different trace elements. A recently developed ion-exchange technique combined with gamma spectrometry has been used, which permits simultaneous determination of a large number of trace elements. The following trace elements have been determined quantitatively: Ag, As, Au, Ba, Br; Ca, Cd, Ce, Co, Cr, Cs, Cu, Fe, Hg, La, Mo, Pt, Rb, Sb, Se, Se, Sm, Zn, W. In some heart samples, Hf and Os were determined qualitatively. The mean and standard deviation are given for the elements Cu, Fe, Se and Zn, Since none of the other quantitatively determined trace elements were normally distributed, the median is given as the central value. When possible, comparisons with values from other investigations have been made. No marked differences in the trace-element concentrations with age or sex could be detected

  9. A RESEARCH AIMED AT DETERMINATION BETWEEN HUMAN RESOURCES PRACTICES AND PERCEIVED ORGANIZATIONAL SUPPORT RELATIONSHIP IN ORGANIZATIONS

    Directory of Open Access Journals (Sweden)

    EBRU AYKAN

    2013-06-01

    Full Text Available Perceived Organizational Support (POS which was popularized in the early 1990s, is conception that may have both positive and negative effect on the staff and organization. In many ways perceived organizational support can determine the continuity of an organization over the long term. This study look at relationship between human resources practices which is taken on five dimensions and perceived organizational support. An investigation has been conduct over bed and supplier industry in Kayseri. The research that was performed with 227 worker is concluded that there are positive relations between training and human resources politics practices and perceived organizational support as of dimensions and between human resource management practices and perceived organizational support as of general.

  10. Mutation Rate Variation is a Primary Determinant of the Distribution of Allele Frequencies in Humans.

    Directory of Open Access Journals (Sweden)

    Arbel Harpak

    2016-12-01

    Full Text Available The site frequency spectrum (SFS has long been used to study demographic history and natural selection. Here, we extend this summary by examining the SFS conditional on the alleles found at the same site in other species. We refer to this extension as the "phylogenetically-conditioned SFS" or cSFS. Using recent large-sample data from the Exome Aggregation Consortium (ExAC, combined with primate genome sequences, we find that human variants that occurred independently in closely related primate lineages are at higher frequencies in humans than variants with parallel substitutions in more distant primates. We show that this effect is largely due to sites with elevated mutation rates causing significant departures from the widely-used infinite sites mutation model. Our analysis also suggests substantial variation in mutation rates even among mutations involving the same nucleotide changes. In summary, we show that variable mutation rates are key determinants of the SFS in humans.

  11. Structural Stereochemistry of Androstene Hormones Determines Interactions with Human Androgen, Estrogen, and Glucocorticoid Receptors

    Directory of Open Access Journals (Sweden)

    Thomas L. Shaak

    2013-01-01

    Full Text Available DHEA, 17α-AED, 17β-AED, and 17β-AET exhibit strong biological activity that has been attributed to androgenic, estrogenic, or antiglucocorticoid activity in vivo and in vitro. This study compared DHEA, 17α-AED, 17β-AED, and 17β-AET for their ability to activate the human AR, ER, and GR and determine the relative androgenicity, estrogenicity, and glucocorticoid activity. The results show that, at the receptor level, these androstene hormones are weak AR and even weaker ER activators. Direct androstene hormone activation of the human AR, ERα, and ERβ may not be essential for their biological function. Similarly, these hormones indirectly activated the human GR, only in the presence of high dexamethasone concentrations. These results underscore the major difference between androstene hormone interactions with these nuclear receptors and their biological effects.

  12. Binding of radioiodinated human. beta. -endorphin to serum proteins from rats and humans, determined by several methods

    Energy Technology Data Exchange (ETDEWEB)

    Sato, H.; Sugiyama, Y.; Sawada, Y.; Iga, T.; Hanano, M.

    1985-10-07

    Binding of immunoreactive radioiodinated human ..beta..-endorphin (/sup 125/I-..beta..-EP) to rat serum was demonstrated by gel filtration of /sup 125/I-..beta..-EP in pooled rat serum on Sephadex G-200. Two radioactive peaks associated with proteins eluted from the column. The first peak eluted at the void volume containing lipoproteins, ..cap alpha../sub 2/- and ..beta../sub 2/-macroglobulins, and the second peak at the fraction of albumin. Binding of /sup 125/I-..beta..-EP to albumin was directly proved by gel filtration of /sup 125/I-..beta..-EP in buffer containing 4% human serum albumin on Sephadex G-200. Equilibrium dialysis was not applicable to investigating the interaction of /sup 125/I-..beta..-EP with serum proteins, because of the intense nonspecific adsorption to the semi-permeable membrane and the degradation of the peptide during dialysis. Therefore, in order to quantitatively evaluate the binding of /sup 125/I-..beta..-EP in sera from rats and humans, the authors utilized four other methods (ultrafiltration, charcoal adsorption, polyethylene glycol precipitation and equilibrium gel filtration). These methods corresponded well with each other and indicated 35-44% binding of /sup 125/I-..beta..-EP in rat serum. Binding of /sup 125/I-..beta..-EP in normal human serum was 36%, determined by ultrafiltration. Serum protein binding of /sup 125/I-..beta..-EP was concentration independent over the concentration range studied (1-1000 nM). 23 references, 4 figures, 1 table.

  13. Determination of mercury in human serum and packed blood cells by neutron activation analysis

    International Nuclear Information System (INIS)

    Versieck, J.; Vanballenberghe, L.; Wittoek, A.; Vermeir, G.; Vandecasteele, C.

    1990-01-01

    A method is described for the determination of mercury in human blood serum and packed blood cells employing neutron activation analysis. Great attention was devoted to the collection and manipulation of the samples. The accuracy and precision of the method were tested by analyzing biological reference materials and by comparing the concentrations measured in a number of serum samples to those obtained by another, independent technique (cold vapor atomic absorption spectrometry) in the same samples. The article reports the levels measured in blood serum and packed blood cells samples from 15 adult volunteers, as well as the figures determined in a open-quotes second-generationclose quotes biological reference material (freeze-dried human serum), prepared and conditioned at the University of Ghent

  14. Determination of flurbiprofen in human plasma by gas chromatography with mass spectrometry and its pharmacokinetics.

    Science.gov (United States)

    Yilmaz, Bilal; Sahin, Huseyin; Akba, Vedat; Erdem, Ali Fuat

    2014-01-01

    This paper describes a GCIMS method for the determination of flurbiprofen in human plasma. Flurbiprofen and internal standard ibuprofen were extracted from plasma by using a liquid-liquid extraction method. Derivatization was carried out using N-Methyl-N-(trimethylsilyl)trifluoroacetamide. The calibration curve was linear between the concentration range of 0.10 and 5.0 microg/mL. Intraday and interday precision values for flurbiprofen in plasma were less than 5.49%, and accuracy (relative error) was better than 5.33%. The extraction recoveries of flurbiprofen from human plasma were between 93.6 and 98.6%. The LOD and LOQ of flurbiprofen were 0.03 and 0.10 microg/mL, respectively. This assay was applied to determine the pharmacokinetic parameters of flurbiprofen in healthy Turkish volunteers who had been given 100 mg of flurbiprofen.

  15. Human error considerations and annunciator effects in determining optimal test intervals for periodically inspected standby systems

    International Nuclear Information System (INIS)

    McWilliams, T.P.; Martz, H.F.

    1981-01-01

    This paper incorporates the effects of four types of human error in a model for determining the optimal time between periodic inspections which maximizes the steady state availability for standby safety systems. Such safety systems are characteristic of nuclear power plant operations. The system is modeled by means of an infinite state-space Markov chain. Purpose of the paper is to demonstrate techniques for computing steady-state availability A and the optimal periodic inspection interval tau* for the system. The model can be used to investigate the effects of human error probabilities on optimal availability, study the benefits of annunciating the standby-system, and to determine optimal inspection intervals. Several examples which are representative of nuclear power plant applications are presented

  16. Fluorine determination in human and animal bones by particle-induced gamma-ray emission

    International Nuclear Information System (INIS)

    Sastri, Chaturvedula S.; Hoffmann, Peter; Ortner, Hugo M.; Iyengar, Venkatesh; Blondiaux, Gilbert; Tessier, Yves; Petri, Hermann; Aras, Namik K.; Zaichick, Vladimir

    2002-01-01

    Fluorine was determined in the iliac crest bones of patients and in ribs collected from postmortem investigations by particle-induced gamma-ray emission based on the 19 F(p,pγ) 19 F reaction, using 20/2.5 MeV protons. The results indicate that for 68% of the human samples the F concentration is in the range 500-1999 μg g -1 . For comparison purposes fluorine was also determined in some animal bones; in some animal tissues lateral profiles of fluorine were measured. (abstract)

  17. Determination of mercury in human tissues after acute poisoning by neutron activation

    International Nuclear Information System (INIS)

    Rakovic, M.; Glagolicova, A.; Prouza, Z.; Gregora, Z.

    1976-01-01

    The non-destructive determination of mercury in human tissues after acute poisoning based on the use of a γ-ray spectometer with a Ge(Li) semiconductor detector is described. Samples were irradiated at a thermal neutron flux density of about 10 12 n cm -2 s -1 for 4 hrs. From the results of the preliminary experiments the following conclusions have been drawn. The mercury losses should be negligible when drying of the biological samples at 80 deg C. To irradiate samples for mercury determination is preferable in silica tubes than in polyethylene ones. (T.I.)

  18. Determination of cobalt in human liver by atomic absorption spectrometry with electrothermal atomization

    International Nuclear Information System (INIS)

    Caldas, E.D.; Gine-Rosias, M.F.; Dorea, J.G.

    1991-01-01

    A detailed study of the use of electrothermal atomic absorption spectrometry for the determination of cobalt in human liver is described. Comparisons of sample digestion using nitric acid or nitric acid plus perchloric acid, atomization procedures and the application of palladium and magnesium nitrate chemical modifiers were studied using NBS SRM 1577a Bovine Liver. The best results were achieved with sample decomposition in nitric acid, atomization from the tube wall and no chemical modifier. Cobalt was determined in 90 samples of livers from foetuses and deceased newborns using the standard addition method with an average recovery of 99.8%. (author). 30 refs.; 4 figs.; 2 tabs

  19. Extraction and Determination of Cyproheptadine in Human Urine by DLLME-HPLC Method

    OpenAIRE

    Maham, Mehdi; Kiarostami, Vahid; Waqif-Husain, Syed; Abroomand-Azar, Parviz; Tehrani, Mohammad Saber; Khoeini Sharifabadi, Malihe; Afrouzi, Hossein; Shapouri, MahmoudReza; Karami-Osboo, Rouhollah

    2013-01-01

    Novel dispersive liquid-liquid microextraction (DLLME), coupled with high performance liquid chromatography with photodiode array detection (HPLC-DAD) has been applied for the extraction and determination of cyproheptadine (CPH), an antihistamine, in human urine samples. In this method, 0.6 mL of acetonitrile (disperser solvent) containing 30 ?L of carbon tetrachloride (extraction solvent) was rapidly injected by a syringe into 5 mL urine sample. After centrifugation, the sedimented phase con...

  20. Direct microcomputer controlled determination of zinc in human serum by flow injection atomic absorption spectrometry

    DEFF Research Database (Denmark)

    Simonsen, Kirsten Wiese; Nielsen, Bent; Jensen, Arne

    1986-01-01

    A procedure is described for the direct determination of zinc in human serum by fully automated, microcomputer controlled flow injection atomic absorption spectrometry (Fl-AAS). The Fl system is pumpless, using the negative pressure created by the nebuliser. It only consists of a three-way valve......, programmable from the microcomputer, to control the sample volume. No pre-treatment of the samples is necessary. The limit of detection is 0.14 mg l–1, and only small amounts of serum (

  1. Carriers of the TCF7L2 rs7903146 TT genotype have elevated levels of plasma glucose, serum proinsulin and plasma gastric inhibitory polypeptide (GIP) during a meal test

    DEFF Research Database (Denmark)

    Gjesing, A P; Kjems, L L; Vestmar, M A

    2011-01-01

    , proinsulin, insulin, C-peptide, glucagon, glucagon-like peptide-1 (GLP-1), glucagon-like peptide-2 (GLP-2) and gastric inhibitory polypeptide (GIP) among individuals carrying the high-risk rs7903146 TT genotype and low-risk CC genotype following a meal test. Methods A meal challenge was performed in 31......, 45, 60, 75, 90, 105, 120, 135, 150, 180, 210, and 240 min after ingestion of a standardised breakfast meal. Results An elevated incremental AUC for plasma glucose was observed among TT genotype carriers (CC carriers 21.8¿±¿101.9 mmol/l¿×¿min vs TT carriers 97.9¿±¿89.2 mmol/l¿×¿min, p¿=¿0.001). TT...

  2. Determination of combined sibship indices "gray zone" using 15 STR loci for central Bosnian human population.

    Science.gov (United States)

    Musanovic, Jasmin; Filipovska-Musanovic, Marijana; Kovacevic, Lejla; Buljugic, Dzenisa; Dzehverovic, Mirela; Avdic, Jasna; Marjanovic, Damir

    2012-05-01

    In our previous population studies of Bosnia and Herzegovina human population, we have used autosomal STR, Y-STR, and X-STR loci, as well as Y-chromosome NRY biallelic markers. All obtained results were included in Bosnian referent database. In order of future development of applied population molecular genetics researches of Bosnia and Herzegovina human population, we have examined the effectiveness of 15 STR loci system in determination of sibship by using 15 STR loci and calculating different cut-off points of combined sibship indices (CSI) and distribution of sharing alleles. From the perspective of its application, it is very difficult and complicated to establish strict CSI cut-off values for determination of the doubtless sibship. High statistically significant difference between the means of CSI values and in distribution of alleles sharing in siblings and non-siblings was noticed (P < 0.0001). After constructing the "gray zone", only one false positive result was found in three CSI cut-off levels with the highest percent of determined sibship/non-sibship at the CSI = 0.067, confirming its practical benefit. Concerning the distribution of sharing alleles, it is recommended as an informative estimator for its usage within Bosnia and Herzegovina human population.

  3. Computational determination of the effects of virulent Escherichia coli and salmonella bacteriophages on human gut.

    Science.gov (United States)

    Mostafa, Marwa Mostafa; Nassef, Mohammad; Badr, Amr

    2016-10-01

    Salmonella and Escherichia coli are different types of bacteria that cause food poisoning in humans. In the elderly, infants and people with chronic conditions, it is very dangerous if Salmonella or E. coli gets into the bloodstream and then they must be treated by phage therapy. Treating Salmonella and E. coli by phage therapy affects the gut flora. This research paper presents a system for detecting the effects of virulent E. coli and Salmonella bacteriophages on human gut. A method based on Domain-Domain Interactions (DDIs) model is implemented in the proposed system to determine the interactions between the proteins of human gut bacteria and the proteins of bacteriophages that infect virulent E. coli and Salmonella. The system helps gastroenterologists to realize the effect of injecting bacteriophages that infect virulent E. coli and Salmonella on the human gut. By testing the system over Enterobacteria phage 933W, Enterobacteria phage VT2-Sa and Enterobacteria phage P22, it resulted in four interactions between the proteins of the bacteriophages that infect E. coli O157:H7, E. coli O104:H4 and Salmonella typhimurium and the proteins of human gut bacterium strains. Several effects were detected such as: antibacterial activity against a number of bacterial species in human gut, regulation of cellular differentiation and organogenesis during gut, lung, and heart development, ammonia assimilation in bacteria, yeasts, and plants, energizing defense system and its function in the detoxification of lipopolysaccharide, and in the prevention of bacterial translocation in human gut. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  4. HAMLET -Human Model MATROSHKA for Radiation Exposure Determination of Astronauts -Current status and results

    Science.gov (United States)

    Reitz, Guenther; Berger, Thomas; Bilski, Pawel; Burmeister, Soenke; Labrenz, Johannes; Hager, Luke; Palfalvi, Jozsef K.; Hajek, Michael; Puchalska, Monika; Sihver, Lembit

    The exploration of space as seen in specific projects from the European Space Agency (ESA) acts as groundwork for human long duration space missions. One of the main constraints for long duration human missions is radiation. The radiation load on astronauts and cosmonauts in space (as for the ISS) is a factor of 100 higher than the natural radiation on Earth and will further increase should humans travel to Mars. In preparation for long duration space missions it is important to evaluate the impact of space radiation in order to secure the safety of the astronauts and minimize their radiation risks. To determine the radiation risk on humans one has to measure the radiation doses to radiosensitive organs within the human body. One way to approach this is the ESA facility MATROSHKA (MTR), under the scientific and project lead of DLR. It is dedicated to determining the radiation load on astronauts within and outside the International Space Station (ISS), and was launched in January 2004. MTR is currently preparing for its fourth experimental phase inside the Japanese Experimental Module (JEM) in summer 2010. MTR, which mimics a human head and torso, is an anthropomorphic phantom containing over 6000 radiation detectors to determine the depth dose and organ dose distribution in the body. It is the largest international research initiative ever performed in the field of space dosimetry and combines the expertise of leading research institutions around the world, thereby generating a huge pool of data of potentially immense value for research. Aiming at optimal scientific exploitation, the FP7 project HAMLET aims to process and compile the data acquired individually by the participating laboratories of the MATROSHKA experiment. Based on experimental input from the MATROSHKA experiment phases as well as on radiation transport calculations, a three-dimensional model for the distribution of radiation dose in an astronaut's body will be built up. The scientific achievements

  5. Determination of extinction coefficients of human hemoglobin in various redox states.

    Science.gov (United States)

    Meng, Fantao; Alayash, Abdu I

    2017-03-15

    The role of hemoglobin (Hb) redox forms in tissue and organ toxicities remain ambiguous despite the well-documented contribution of Hb redox reactivity to cellular and subcellular oxidative changes. Moreover, several recent studies, in which Hb toxicity were investigated, have shown conflicting outcomes. Uncertainties over the potential role of these species may in part be due to the protein preparation method of choice, the use of published extinction coefficients and the lack of suitable controls for Hb oxidation and heme loss. Highly purified and well characterized redox forms of human Hb were used in this study and the extinction coefficients of each Hb species (ferrous/oxy, ferric/met and ferryl) were determined. A new set of equations were established to improve accuracy in determining the transient ferryl Hb species. Additionally, heme concentrations in solutions and in human plasma were determined using a novel reversed phase HPLC method in conjugation with our photometric measurements. The use of more accurate redox-specific extinction coefficients and method calculations will be an invaluable tool for both in vitro and in vivo experiments aimed at determining the role of Hb-mediated vascular pathology in hemolytic anemias and when Hb is used as oxygen therapeutics. Published by Elsevier Inc.

  6. Apoptosis may determine the release of skeletal alkaline phosphatase activity from human osteoblast-line cells.

    Science.gov (United States)

    Farley, J R; Stilt-Coffing, B

    2001-01-01

    Although quantitative measurement of skeletal alkaline phosphatase (sALP) activity in serum can provide an index of the rate of bone formation, the metabolic process that determines the release of sALP - from the surface of osteoblasts, into circulation-is unknown. The current studies were intended to examine the hypothesis that the release of sALP from human osteoblasts is a consequence of apoptotic cell death. We measured the release of sALP activity from human osteosarcoma (SaOS-2) cells and normal human bone cells, under basal conditions and in response to agents that increased apoptosis (TNF-a, okadiac acid) and agents that inhibit apoptosis (IGF-I, calpain, and caspase inhibitors). Apoptosis was determined by the presence of nucleosomes (histone-associated DNA) in the cytoplasm of the cells by using a commercial kit. The results of these studies showed that TNF-a and okadiac acid caused dose- and time-dependent increases in apoptosis in the SaOS-2 cells (r = 0.78 for doses of TNF-a and r = 0.93 for doses of okadiac acid, P sALP activity (e.g., r = 0.89 for TNF-a and r = 0.75 for okadiac acid, P sALP activity (P sALP activity (P sALP release. The associations between apoptosis and sALP release were not unique to osteosarcoma (i.e., SaOS-2) cells, but also seen with osteoblast-line cells derived from normal human bone. Together, these data demonstrate that the release of sALP activity from human osteoblast-line cells in vitro is associated with, and may be a consequence of, apoptotic cell death. These findings are consistent with the general hypothesis that the appearance of sALP activity in serum may reflect the turnover of osteoblast-line cells.

  7. Adsorptive Cathodic Stripping Voltammetric Determination of Cefoperazone in Bulk Powder, Pharmaceutical Dosage Forms, and Human Urine

    Directory of Open Access Journals (Sweden)

    Vu Dang Hoang

    2013-01-01

    Full Text Available The electroreduction behaviour and determination of cefoperazone using a hanging mercury drop electrode were investigated. Cyclic voltammograms of cefoperazone recorded in universal Britton-Robinson buffers pH 3–6 exhibited a single irreversible cathodic peak. The process was adsorption-controlled. Britton-Robinson buffer 0.04 M pH 4.0 was selected as a supporting electrolyte for quantitative purposes by differential pulse and square wave adsorptive cathodic stripping voltammetry. The experimental voltammetric conditions were optimized using Central Composite Face design. A reduction wave was seen in the range from −0.7 to −0.8 V. These voltammetric techniques were successfully validated as per ICH guidelines and applied for the determination of cefoperazone in its single and sulbactam containing powders for injection and statistically comparable to USP-HPLC. They were further extended to determine cefoperazone in spiked human urine with no matrix effect.

  8. Differences in Virulence Between Legionella pneumophila Isolates From Human and Non-human Sources Determined in Galleria mellonella Infection Model

    Directory of Open Access Journals (Sweden)

    Patrícia S. Sousa

    2018-04-01

    Full Text Available Legionella pneumophila is a ubiquitous bacterium in freshwater environments and in many man-made water systems capable of inducing pneumonia in humans. Despite its ubiquitous character most studies on L. pneumophila virulence focused on clinical strains and isolates from man-made environments, so little is known about the nature and extent of virulence variation in strains isolated from natural environments. It has been established that clinical isolates are less diverse than man-made and natural environmental strains, suggesting that only a subset of environmental isolates is specially adapted to infect humans. In this work we intended to determine if unrelated L. pneumophila strains, isolated from different environments and with distinct virulence-related genetic backgrounds, displayed differences in virulence, using the Wax Moth Galleria mellonella infection model. We found that all tested strains were pathogenic in G. mellonella, regardless of their origin. Indeed, a panoply of virulence-related phenotypes was observed sustaining the existence of significant differences on the ability of L. pneumophila strains to induce disease. Taken together our results suggest that the occurrence of human infection is not related with the increased capability of some strains to induce disease since we also found a concentration threshold above which L. pneumophila strains are equally able to cause disease. In addition, no link could be established between the sequence-type (ST and L. pneumophila pathogenicity. We envision that in man-made water distribution systems environmental filtering selection and biotic competition acts structuring L. pneumophila populations by selecting more resilient and adapted strains that can rise to high concentration if no control measures are implemented. Therefore, public health strategies based on the sequence based typing (STB scheme analysis should take into account that the major disease-associated clones of L

  9. The determination of methylmercury, total mercury and total selenium in human hair

    International Nuclear Information System (INIS)

    1987-01-01

    This Reference Method describes the determination of methylmercury in human hair by gas liquid chromatography. It is designed for biological monitoring of selected individuals and population groups with a possible intake of methylmercury exceeding the recommended Provisional Tolerable Weekly Intake (PTWI) through contaminated seafood, as part of a project on the evaluation of methylmercury in Mediterranean populations and related health hazards. The method, however, is also applicable to other regions. The method involves direct determination of methylmercury by gas liquid chromatography. The sample is disintegrated in a solution of sodium hydroxide, methylmercury is extracted from an aliquot of the solution into toluene and, after purification, a small volume is injected into a chromatographic column, filled with polyethyleneglycol succinate on Diatomite AW. Methylmercury in the gaseous mixture is detected with an electron capture detector and its amount determined by comparing the peak height with those of appropriate standards. The next Reference Method describes the determination of selenium in human hair (and other indicative tissues) by gas liquid chromatography and is designed for biological monitoring of selected individuals and population groups in the Mediterranean region with a possible intake of methylmercury exceeding the recommended Provisional Tolerable Weekly Intake (PTWI) through contaminated seafood. The data are intended to establish a possible correlation between methylmercury intake and levels of selenium in the subjects monitored. Selenium in the solvent phase is determined by gas liquid chromatography using an electron capture detector. The above method has been selected because selenium is determined in conjunction with methylmercury, both of which require competence in gas chromatographic techniques. Reliable result for total selenium, however, will also be obtained by the following techniques: a) Atomic absorption spectrophotometry; b

  10. Experimental Determination of the Neutron Radiation-Dose Distribution in the Human Phantom

    Energy Technology Data Exchange (ETDEWEB)

    Stipcic, Neda [Institute Rudjer Bogkovic, Zagreb, Yugoslavia (Serbia)

    1967-01-15

    The quality of the radiation delivering the radiation dose to the human phantom is quite different from that of the incident neutron beam. This paper describes the experimental investigation of the variation of neutron dose related to the variation of neutron fluence with depth in the human phantom. The distribution of neutron radiation was determined in the human phantom - a cube of paraffin wax 25 cm x 25 cm x 50 cm with a density of 0.92 cm{sup -3}. Po-Be and Ra-Be point sources were used as neutron sources. Neutron fluences were measured using different types of detector: scintillation detector, BF{sub 3} counter, and nuclear-track emulsions. Since the fluence measurements with these three types of detectors were carried out under the same experimental conditions, it was possible to separate and analyse each part of the radiation dose in the paraffin. From the investigations, the distribution of the total radiation dose was obtained as a function of the paraffin depth. The maximum value of this dose distribution is constant with respect to the distance between the source and the paraffin phantom. From the results obtained, some conclusions may be drawn concerning the amount of absorbed radiation dose in the human phantom. (author)

  11. Major differences between human atopic dermatitis and murine models, as determined by using global transcriptomic profiling

    DEFF Research Database (Denmark)

    Ewald, David A.; Noda, Shinji; Oliva, Margeaux

    2017-01-01

    , and a comparison of these models with the human AD transcriptomic fingerprint is lacking. Objective We sought to evaluate the transcriptomic profiles of 6 common murine models and determine how they relate to human AD skin. Methods Transcriptomic profiling was performed by using microarrays and quantitative RT......-PCR on biopsy specimens from NC/Nga, flaky tail, Flg-mutated, ovalbumin-challenged, oxazolone-challenged, and IL-23–injected mice. Gene expression data of patients with AD, psoriasis, and contact dermatitis were obtained from previous patient cohorts. Criteria of a fold change of 2 or greater and a false...... discovery rate of 0.05 or less were used for gene arrays. Results IL-23–injected, NC/Nga, and oxazolone-challenged mice show the largest homology with our human meta-analysis–derived AD transcriptome (37%, 18%, 17%, respectively). Similar to human AD, robust TH1, TH2, and also TH17 activation are seen in IL...

  12. Determination of Chlorpyrifos in Human Blood by Gas Chromatography-Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Xinhua Dai

    2017-01-01

    Full Text Available Gas chromatography-mass spectrometry method was developed for the qualitative and quantitative analyses of chlorpyrifos in human blood samples. The chlorpyrifos and parathion (internal standard in human blood were extracted with a mixed solvent of hexane and acetonitrile. Chlorpyrifos was well separated from the internal standard. The linear range of chlorpyrifos was 0.01–2 μg/ml in blood. The limit of detection and limit of quantification were estimated at 0.002 and 0.01 μg/ml, respectively. The inter- and intra-day precisions, accuracy, and recovery were assessed to verify this method. The results showed that the developed method is rapid, sensitive, and reliable. It is suitable for the determination of chlorpyrifos in forensic toxicological analysis and clinical diagnosis.

  13. Determination of the synthesis of uptake of α2-macroglobulin by cultured human glioma cells

    International Nuclear Information System (INIS)

    Druskova, E.; Bizik, J.; Grofova, M.

    1994-01-01

    Using immunological techniques, the synthesis of α 2 -macroglobulin was studied in established cell lines derived from human glioblastomas multiform. α 2 -Macroglobulin was detected in cytoplasm and in the culture medium of the analyzed cell lines. Radioimmunoprecipitation, revealed a protein with Mr corresponding to α 2 -macroglobulin in the medium conditioned by U-118MG and U-343MG cells. On the other hand, using immunoblot analysis, α 2 -macroglobulin was detected in all of the analyzed lines. In immunofluorescence test, α 2 -macroglobulin was determined also in all four cell lines, but with different staining pattern. Conditioned culture medium of U-536MG cells with the lowest level of α 2 -macroglobulin exerted the lowest mitogenic activity for human fibroblasts. (author)

  14. Study on the Analytical Method for Determination of P-32 in Human Hair

    International Nuclear Information System (INIS)

    Syarbaini; Lubis, E.; Sarwani

    1996-01-01

    Neutron doses due to accident criticality can be estimated by measuring of radionuclide of neutron activation products in human hair. In this work, the analytical method for the determination of P-32 in neutron irradiated hair sample by G.A Siwabessy reactor has been studied. This analytical method consists of dissolving of human hair sample by 10 M HNO3, separation dan purification of P-32 by precipitation as ammonium molibdophosphate finally, the precipitate was measured by low backgroundα/βcounter. The minimum detectable activity of P-32 was 0,05 Bq at a background of 4,6 cpm and with a counting efficiency of 55 % for a 30 minute counting time

  15. Determination of albendazole sulfoxide in human plasma by using liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Saraner, Nihal; Özkan, Güler Yağmur; Güney, Berrak; Alkan, Erkin; Burul-Bozkurt, Nihan; Sağlam, Onursal; Fikirdeşici, Ezgi; Yıldırım, Mevlüt

    2016-06-01

    A rapid, simple and sensitive method was developed and validated using liquid chromatography-tandem mass spectrometry (LC-MS/MS) for determination of albendazole sulfoxide (ABZOX) in human plasma. The plasma samples were extracted by protein precipitation using albendazole sulfoxide-d3 as internal standard (IS). The chromatographic separation was performed on Waters Xbridge C18Column (100×4.6mm, 3.5μm) with a mobile phase consisting of ammonia solution, water and methanol at a flow rate of 0.70mL/min. ABZOX was detected and identified by mass spectrometry with electrospray ionization (ESI) in positive ion and multiple-reaction monitoring (MRM) mode. The method was linear in the range of 3-1500ng/mL for ABZOX. This method was successfully applied to the bioequivalence study in human plasma samples. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Essential role of RAB27A in determining constitutive human skin color.

    Directory of Open Access Journals (Sweden)

    Yasuko Yoshida-Amano

    Full Text Available Human skin color is predominantly determined by melanin produced in melanosomes within melanocytes and subsequently distributed to keratinocytes. There are many studies that have proposed mechanisms underlying ethnic skin color variations, whereas the processes involved from melanin synthesis in melanocytes to the transfer of melanosomes to keratinocytes are common among humans. Apart from the activities in the melanogenic rate-limiting enzyme, tyrosinase, in melanocytes and the amounts and distribution patterns of melanosomes in keratinocytes, the abilities of the actin-associated factors in charge of melanosome transport within melanocytes also regulate pigmentation. Mutations in genes encoding melanosome transport-related molecules, such as MYO5A, RAB27A and SLAC-2A, have been reported to cause a human pigmentary disease known as Griscelli syndrome, which is associated with diluted skin and hair color. Thus we hypothesized that process might play a role in modulating skin color variations. To address that hypothesis, the correlations of expression of RAB27A and its specific effector, SLAC2-A, to melanogenic ability were evaluated in comparison with tyrosinase, using human melanocytes derived from 19 individuals of varying skin types. Following the finding of the highest correlation in RAB27A expression to the melanogenic ability, darkly-pigmented melanocytes with significantly higher RAB27A expression were found to transfer significantly more melanosomes to keratinocytes than lightly-pigmented melanocytes in co-culture and in human skin substitutes (HSSs in vivo, resulting in darker skin color in concert with the difference observed in African-descent and Caucasian skins. Additionally, RAB27A knockdown by a lentivirus-derived shRNA in melanocytes concomitantly demonstrated a significantly reduced number of transferred melanosomes to keratinocytes in co-culture and a significantly diminished epidermal melanin content skin color intensity (

  17. Scintillometric determination of DNA repair in human cell lines. A critical appraisal

    Energy Technology Data Exchange (ETDEWEB)

    Bianchi, V.; Zantedeschi, A.; Levis, A.G. (Padua Univ. (Italy). Ist. di Biologica Animale); Nuzzo, F.; Stefanini, M. (Consiglio Nazionale delle Ricerche, Pavia (Italy). Ist. di Genetica Biochimica ed Evoluzionistica); Abbondandolo, A.; Bonatti, S.; Fiorio, R.; Mazzaccaro, A. (Consiglio Nazionale delle Ricerche, Pisa (Italy). Ist. di Mutagenesi e Differenziamento); Capelli, E. (Pavia Univ. (Italy). Ist. di Genetica)

    1982-04-01

    The ability of a variety of chemical and physical agents to stimulate DNA repair synthesis in human cell cultures was tested by a simplified scintillometric procedure, with the use of hydroxyurea (HU) to suppress DNA replicative synthesis. After incubation with (/sup 3/H)thymidine, the radioactivity incorporated into DNA was determined in controls (C) and treated (T) cultures and in the corresponding HU series (Csub(HU), Tsub(HU)). The ratios Tsub(HU)/Csub(HU) and Tsub(HU)/T:Csub(HU)/C, indicating absolute and relative increases of DNA radioactivity, were calculated. When both ratios were significantly higher than 1, they were taken as indices of DNA repair stimulation.

  18. Determination of arsenic in human hair by destructive neutron activation analysis

    Energy Technology Data Exchange (ETDEWEB)

    Bayat, I.; Ansar Kabootar Ahangi, M.; Sadegh Keramati, M. [Atomic Energy Organization of Iran, Waste Management Dept., Tehran (Iran)

    2002-03-01

    The method described below was developed for the purposes of the determination of micro quantities of arsenic in human hair. The nuclear research reactor of the AEOI, the Atomic Energy Organization of Iran, was used for the irradiation of 150 mg hair samples for a period of 20 hours. Radiochemical methods were used for the separation of interfering ions. The total arsenic concentrations found in over 100 samples ranged from 0.01 to 4 ppm, with the detection limit for arsenic being reported to lie between 0.1 and 0.01 ppm. (orig.)

  19. Women’s human agency and self-determination in Guatemalan tourism development

    Directory of Open Access Journals (Sweden)

    Lindsay Usher

    2010-10-01

    Full Text Available Tourism is often identified as a strategy to ameliorate the wellbeing of poor communities and their most vulnerable members – women and their children. Women’s ability to get involved in and benefit from tourism is, however, conditioned by traditional gender roles and consequent education handicaps. Development programs often target women to mitigate these disparities. Thispaper examines whether an intervention by the Peace Corps resulted in improved human agency and self-determination among indigenous Q’eqchi’ women in Alta Verapaz, Guatemala. The paper reports the accomplishments and shortcomings of the intervention and elaborates on the practical and theoretical implications of the findings.

  20. High Performance Liquid Chromatographic Method for Determination of Dipyridamole in Human Plasma

    Directory of Open Access Journals (Sweden)

    DAVOOD BEIGI BAND ARAB ADI

    1999-08-01

    Full Text Available A simple, rapid and specific high-performance liquid chromatographic procedure is reported for"nquantitative determination of dipyridamole in human -plasma. The assay uses a reversed-phase"nhigh-performance liquid chromatographic (HPLC and UV detection at 280nm and has a limit"nof detection of approximately 5ng/mL. The mobile phase consists of MeOH-H20 (60:40"nadjusted to pH 3.3. Dipyridamole was extracted from plasma by back-extraction procedure, with"npropranolol as the internal standard. The reproducibility of the method is satisfactory

  1. Determination of organic-bound chlorine and bromine in human body fluids by neutron activation analysis

    International Nuclear Information System (INIS)

    McKinney, J.D.; Abusamra, A.; Reed, J.H.

    1983-01-01

    The levels of organic-bound chlorine and bromine in human milk and serum are determined by neutron activation analysis. Desalted milk and serum fractions are irradiated with neutrons in a nuclear reactor and the resulting γ-rays of 38 Cl and 80 Br are measured. The desalting procedure, achieved by using Bio-Gel molecular sieves, virtually removes all ionic chloride and bromides from milk and serum. Radioactive tracer studies with polychlorinated biphenyl- 14 C indicate a recovery of 90% through the Bio-Gel column. The total organic chlorine in 2.2-(4-chlorophenyl)-1,1-dichloroethane spiked milk and heptachlor spiked milk, determined after being desalted and irradiated according to this procedure, substantiates a good recovery of the added spike. The lower limits of detection of organic-bound chlorine and bromine in milk or serum are 50 and 5 parts per billion (ppb), respectively

  2. Online determination of biophysical parameters of mucous membranes of a human body

    Energy Technology Data Exchange (ETDEWEB)

    Lisenko, S A; Kugeiko, M M [Belarusian State University, Minsk (Belarus)

    2013-07-31

    We have developed a method for online determination of biophysical parameters of mucous membranes (MMs) of a human body (transport scattering coefficient, scattering anisotropy factor, haemoglobin concentration, degrees of blood oxygenation, average diameter of capillaries with blood) from measurements of spectral and spatial characteristics of diffuse reflection. The method is based on regression relationships between linearly independent components of the measured light signals and the unknown parameters of MMs, obtained by simulation of the radiation transfer in the MM under conditions of its general variability. We have proposed and justified the calibration-free fibre-optic method for determining the concentration of haemoglobin in MMs by measuring the light signals diffusely reflected by the tissue in four spectral regions at two different distances from the illumination spot. We have selected the optimal wavelengths of optical probing for the implementation of the method. (laser applications in biology and medicine)

  3. [Determination of trace cobalt in human urine by graphite furnace atomic absorption spectrometr].

    Science.gov (United States)

    Zhong, L X; Ding, B M; Jiang, D; Liu, D Y; Yu, B; Zhu, B L; Ding, L

    2016-05-20

    To establish a method to determine cobalt in human urine by graphite furnace atomic absorption spectrometry. Urine with 2% nitric acid diluted two-fold, to quantify the curve, graphite furnace atomic absorption spectrometric detection. Co was linear within 2.5~40.0 ng/ml with r>0.999. Spike experiment showed that Co received good recovery rate, which was 90.8%~94.8%. Intra-assay precisions were 3.2%~5.1% for Co, inter-assay precisions were 4.4%~5.2% for Co. The method by using graphite furnace atomic absorption spectrometr to determine urine Co was fast, accurate and with low matrix effect. It could meet the requirement in GBZ/T 210.5-2008.

  4. From genomes to societies: a holistic view of determinants of human health.

    Science.gov (United States)

    Shi, Yuyan; Zhong, Sheng

    2014-08-01

    Both biological and social sciences have identified contributing factors to human health. However, health outcomes are unlikely to equal a simple sum of these identified factors. This article makes an attempt to put together the information, methods, and technologies that relate to health outcomes from biological, behavioral, and social disciplines. Much of this information was obtained by controlling for the variations of the factors in 'other' disciplines. For example, genetic factors were controlled for in identifying the behavioral determinants of health. Looking forward, better understandings of health outcomes may require exploiting the interactions of health determinants that were identified from different disciplines. We propose the concept of 'systems health' studies, which take health outcomes as the outputs of a system, where the inputs and their interactions from multiple disciplines are considered. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. Molecular Determinants of Human T-lymphotropic Virus Type 1 Transmission and Spread

    Directory of Open Access Journals (Sweden)

    Patrick L. Green

    2011-07-01

    Full Text Available Human T-lymphotrophic virus type-1 (HTLV-1 infects approximately 15 to 20 million people worldwide, with endemic areas in Japan, the Caribbean, and Africa. The virus is spread through contact with bodily fluids containing infected cells, most often from mother to child through breast milk or via blood transfusion. After prolonged latency periods, approximately 3 to 5% of HTLV-1 infected individuals will develop either adult T-cell leukemia/lymphoma (ATL, or other lymphocyte-mediated disorders such as HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP. The genome of this complex retrovirus contains typical gag, pol, and env genes, but also unique nonstructural proteins encoded from the pX region. These nonstructural genes encode the Tax and Rex regulatory proteins, as well as novel proteins essential for viral spread in vivo such as, p30, p12, p13 and the antisense encoded HBZ. While progress has been made in the understanding of viral determinants of cell transformation and host immune responses, host and viral determinants of HTLV-1 transmission and spread during the early phases of infection are unclear. Improvements in the molecular tools to test these viral determinants in cellular and animal models have provided new insights into the early events of HTLV-1 infection. This review will focus on studies that test HTLV-1 determinants in context to full length infectious clones of the virus providing insights into the mechanisms of transmission and spread of HTLV-1.

  6. Potentiometric determination of acid dissociation constants (pKa) for human and veterinary antibiotics.

    Science.gov (United States)

    Qiang, Zhimin; Adams, Craig

    2004-07-01

    This work determined the acid dissociation constants (pKa) of 26 common human and veterinary antibiotics by potentiometric titration. Selected antibiotics consisted of sulfonamides, macrolides, tetracyclines, fluoroquinolones, and other miscellaneous antibiotics. After validation of analysis methods using phosphoric acid as a model compound, a second-derivative (delta2pH/deltaV2) method was primarily applied to determining pKa's from titration curves for most antibiotics due to its convenience and accuracy. For tetracyclines, however, a least-square non-linear regression method was developed to determine their pKa's because the second-derivative method cannot well distinguish the pKa,2 and pKa,3 of tetracyclines. Results indicate that the pKa values are approximately 2 and 5-7.5 for sulfonamides; 7.5-9 for macrolides; 3-4, 7-8 and 9-10 for tetracyclines; 3-4, 6, 7.5-9 and 10-11 for fluoroquinolones; while compound-specific for other miscellaneous antibiotics. The moieties corresponding to specific pKa's were identified based on chemical structures of antibiotics. In addition, the pKa's available in literature determined by various techniques are compiled in comparison with the values of this work. These results are expected to essentially facilitate the research on occurrence, fate and effects, analysis methods development, and control of antibiotics in various treatment operations.

  7. Establishing a protocol for element determination in human nail clippings by neutron activation analysis

    International Nuclear Information System (INIS)

    Sanches, Thalita Pinheiro; Saiki, Mitiko

    2011-01-01

    Human nail samples have been analyzed to evaluate occupational exposure, nutritional status and to diagnose certain diseases. However, sampling and washing protocols for nail analyses vary from study to study not allowing comparisons between studies. One of the difficulties in analyzing nail samples is to eliminate only surface contamination without removing elements of interest in this tissue. In the present study, a protocol was defined in order to obtain reliable results of element concentrations in human nail clippings. Nail clippings collected from all 10 fingers or toes were previously pre cleaned using an ethyl alcohol solution to eliminate microbes. Then, the clippings were cut in small pieces and submitted to different reagents for washing by shaking. Neutron activation analysis (NAA) was applied for nail samples analysis which consisted of irradiating aliquots of samples together with synthetic elemental standards in the IEA-R1 nuclear research reactor followed by gamma ray spectrometry. Comparisons made between the results obtained for nails submitted to different reagents for cleaning indicated that the procedure using acetone and Triton X100 solution is more effective than that of nitric acid solution. Analyses in triplicates of a nail sample indicated results with relative standard deviations lower than 15% for most of elements, showing the homogeneity of the prepared sample. Qualitative analyses of different nail polishes showed that the presence of elements determined in the present study is negligible in these products. Quality control of the analytical results indicated that the applied NAA procedure is adequate for human nail analysis. (author)

  8. Video Analysis of Human Gait and Posture to Determine Neurological Disorders

    Directory of Open Access Journals (Sweden)

    Ivan Lee

    2008-08-01

    Full Text Available This paper investigates the application of digital image processing techniques to the detection of neurological disorder. Visual information extracted from the postures and movements of a human gait cycle can be used by an experienced neurologist to determine the mental health of the person. However, the current visual assessment of diagnosing neurological disorder is based very much on subjective observation, and hence the accuracy of diagnosis heavily relies on experience. Other diagnostic techniques employed involve the use of imaging systems which can only be operated under highly constructed environment. A prototype has been developed in this work that is able to capture the subject's gait on video in a relatively simple setup, and from which to process the selected frames of the gait in a computer. Based on the static visual features such as swing distances and joint angles of human limbs, the system identifies patients with Parkinsonism from the test subjects. To our knowledge, it is the first time swing distances are utilized and identified as an effective means for characterizing human gait. The experimental results have shown a promising potential in medical application to assist the clinicians in diagnosing Parkinsonism.

  9. [Determination of 24 minerals in human milk by inductively coupled plasma mass spectrometry with microwave digestion].

    Science.gov (United States)

    Sun, Zhongqing; Yue, Bing; Yang, Zhenyu; Li, Xiaowei; Wu, Yongning; Yin, Shian

    2013-05-01

    To determine the levels of 24 minerals in human milk by inductively coupled plasma mass spectrometry with microwave digestion. The samples were digested by microwave. The contents of minerals were determined by inductively coupled plasma mass spectrometry. The standard reference minerals of 1849a and 1568a from National Institute of Science and Technology were used for quality control. The accuracy and reproduability for this method were evaluated with mix standards and 1849a and 1568a standard reference materials. The ranges of the levels of sodium, magnesium, phosphorus, potassium, calcium, aluminum, chromium, arsenic, selenium, iron, zinc, manganese, copper, molybdenum, vanadium, cobalt, nickel, gallium, cadmium, silver, strontium, cesium, barium, lead in human milk was 34.97-415.83 mg/kg, 19.00-39.52 mg/kg, 102.13-274.53 mg/kg, 351.19-713.99 mg/kg, 180.08-349.64 mg/kg, 0.06-0.44 mg/kg, 0.9-7.37 microg/kg, 0.92-2.72 microg/kg, 0.20-21.15 microg/kg, 0.10-0.70 mg/kg, 0.56-3.25 mg/kg, 3.00-16.12 micro.g/kg, 62.16-591.69 microg/kg, 0.02-6.91 microg/kg, 5.99-13.70 microg/kg, 0.07-2.11 microg/kg, 0.77-209.26 microg/kg, 0.005-0.28 microg/kg, 0.02-0.23 microg/kg, 0.02-0.71 microg/kg, 36.89-132.26 microg/kg, 0.01-4.72 microg/kg, 0.83-28.16 microg/kg, 2.5-5.3 microg/kg, respectively. The levels of minerals in human milk in present study were consisted with other similar studies. The experiment examined the levels of minerals in human milk satisfactorily. The method has high accuracy and good reproducibility, which could be used for understanding the levels of minerals in human milk.

  10. Trace element determination study in human hair by neutron activation analysis

    International Nuclear Information System (INIS)

    Frazao, Selma Violato

    2008-01-01

    Human hair analysis studies have been subject of continuous interest due to the fact that they can be used as an important tool to evaluate trace element levels in the human body. These determinations have been carried out to use hair for environmental and occupational monitoring, to identify intoxication or poisoning by toxic metals, to assess nutritional status, to diagnose and to prevent diseases and in forensic sciences. Although hair analysis presents several advantages over other human tissue or fluid analyses, such as organ tissue, blood, urine and saliva, there are some controversies regarding the use of hair analysis data. These controversies arise from the fact that it is difficult to establish reliable reference values for trace elements in hair. The purpose of this study was to evaluate the factors that affect element concentrations in hair samples from a population considered healthy and residing in the Sao Paulo metropolitan area. The collected human head hair was cut in small pieces, washed, dried and analyzed by neutron activation analysis (NAA). Aliquots of hair samples and synthetic elemental standards were irradiated at the IEA-R1 nuclear research reactor for 16 h under a thermal neutron flux of about 5x10 12 n cm -2 s -1 for As, Br, Ca, Co, Cr, Cs, Cu, Fe, K, La, Na, Sb, Sc, Se and Zn determinations. The induced gamma activities of the standards and samples were measured using a gamma ray spectrometer coupled to an hiperpure Ge detector. For quality control of the results, IAEA- 85 Human Hair and INCT-TL-1 Tea Leaves certified reference materials (CRMs) were analyzed. Results obtained in these CRMs presented for most of elements, good agreement with the values of the certificates (relative errors less than 10%) and good precision (variation coefficients less than 13.6%). Results of replicate hair sample analysis showed good reproducibility indicating homogeneity of the prepared sample. Results obtained in the analyses of dyed and non-dyed hair

  11. Determination of Flurbiprofen in Human Plasma by High-Performance Liquid Chromatography.

    Science.gov (United States)

    Yilmaz, Bilal; Erdem, Ali Fuat

    2015-10-01

    A simple high-performance liquid chromatography method has been developed for determination of flurbiprofen in human plasma. The method was validated on an Ace C18 column using UV detection. The mobile phase was acetonitrile-0.05 M potassium dihydrogen phosphate solution (60:40, v/v) adjusted to pH 3.5 with phosphoric acid. The calibration curve was linear between the concentration range of 0.10-5.0 μg/mL. Intra- and inter-day precision values for flurbiprofen in plasma were flurbiprofen from human plasma were between 93.0 and 98.9%. The limits of detection and quantification of flurbiprofen were 0.03 and 0.10 μg/mL, respectively. In addition, this assay was applied to determine the pharmacokinetic parameters of flurbiprofen in six healthy Turkish volunteers who had been given 100 mg flurbiprofen. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  12. Determination of human albumin in serum and urine samples by constant-energy synchronous fluorescence method.

    Science.gov (United States)

    Madrakian, Tayyebeh; Bagheri, Habibollah; Afkhami, Abbas

    2015-08-01

    A sensitive spectrofluorimetric method using constant-energy synchronous fluorescence technique is proposed for the determination of human albumin without separation. In this method, no reagent was used for enhancement of the fluorescence signal of albumin in the solution. Effects of some parameters, such as energy difference between excitation and emission monochromators (ΔE), emission and excitation slit widths and scan rate of wavelength were studied and the optimum conditions were established. For this purpose factorial design and response surface method were employed for optimization of the effective parameters on the fluorescence signal. The results showed that the scan rate of the wavelength has no significant effect on the analytical signal. The calibration curve was linear in the range 0.1-220.0 µg mL(-1) of albumin with a detection limit of 7.0 × 10(-3)  µg mL(-1). The relative standard deviations (RSD) for six replicate measurements of albumin were calculated as 2.2%, 1.7% and 1.3% for 0.5, 10.0 and 100.0 µg mL(-1) albumin, respectively. Furthermore the proposed method has been employed for the determination of albumin in human serum and urine samples. Copyright © 2014 John Wiley & Sons, Ltd.

  13. Determination of cadmium in human urine by graphite furnace atomic absorption spectrometry

    International Nuclear Information System (INIS)

    Shimizu, Tokuo; Shijo, Yoshio; Sakai, Kaoru

    1981-01-01

    A trace amount of cadmium in human urine was determined by graphite furnace atomic absorption spectrometry. A urine sample (25 ml) was digested with 5 ml of HNO 3 and 30 ml of H 2 O 2 in a long-neck flask on a hot-plate (200 0 C), then diluted to 50 ml. The standard addition method was carried out before digesting. Ten μl of the resulted solution was injected into a tube treated with tungsten carbide, and the cadmium signal was measured with the ramp mode atomization. Interference induced by organic materials in urine was avoided by HNO 3 -H 2 O 2 digestion. Interference induced by inorganic salts could be reduced by 2-fold dilution and tungsten carbide treatment. The cadmium signal was separated sufficiently from the molecular absorption due to NaCl etc. by the ramp mode atomization. Since the blank level of H 2 O 2 was relatively high, the determination was limited to about 0.1 μg/l. The coefficient of variation was 1.76% at 0.36 μg/l in 24 h human urine (n = 4). The time required was (8 -- 10)h. The precision of this method was higher than those of direct methods, and the reasonable values of urine levels of cadmium were obtained. (author)

  14. Application of dried blood spot cards to determine olive oil phenols (hydroxytyrosol metabolites) in human blood.

    Science.gov (United States)

    de Las Hazas, María Carmen López; Motilva, Maria José; Piñol, Carme; Macià, Alba

    2016-10-01

    In this study, a fast and simple blood sampling and sample pre-treatment method based on the use of the dried blood spot (DBS) cards and ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) for the quantification of olive oil phenolic metabolites in human blood was developed and validated. After validation, the method was applied to determine hydroxytyrosol metabolites in human blood samples after the acute intake of an olive oil phenolic extract. Using the FTA DMPK-A DBS card under optimum conditions, with 20µL as the blood solution volume, 100µL of methanol/Milli-Q water (50/50, v/v) as the extraction solvent and 7 disks punched out from the card, the main hydroxytyrosol metabolites (hydroxytyrosol-3-O-sulphate and hydroxytyrosol acetate sulphate) were identified and quantified. The developed methodology allowed detecting and quantifying the generated metabolites at low μM levels. The proposed method is a significant improvement over existing methods to determine phenolic metabolites circulating in blood and plasma samples, thus making blood sampling possible with the volunteer pricking their own finger, and the subsequent storage of the blood in the DBS cards prior to chromatographic analysis. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. A novel human body exergy consumption formula to determine indoor thermal conditions for optimal human performance in office buildings

    DEFF Research Database (Denmark)

    Wu, Xiaozhou; Zhao, Jianing; Olesen, Bjarne W.

    2013-01-01

    In this paper, a novel human body exergy consumption formula was derived strictly according to Gagge's two-node thermal transfer model. The human body exergy consumption calculated by the formula was compared with values calculated using Shukuya's formula for a typical office environment....... The results show that human body exergy consumption calculated by either of these formulas reaches a minimum under the same thermal condition. It is shown that this is in accordance with expectation. The relation between human performance and human body exergy consumption was studied by analyzing the data...... obtained in simulated office environments in winter. The results show that human body exergy consumption and human performance are inversely as operative temperature changes from 17 to 28°C or human thermal sensation changes from −1.0 to +1.4, and that optimum thermal comfort cannot be expected to lead...

  16. Pedaling rate is an important determinant of human oxygen uptake during exercise on the cycle ergometer.

    Science.gov (United States)

    Formenti, Federico; Minetti, Alberto E; Borrani, Fabio

    2015-09-01

    Estimation of human oxygen uptake (V˙o2) during exercise is often used as an alternative when its direct measurement is not feasible. The American College of Sports Medicine (ACSM) suggests estimating human V˙o2 during exercise on a cycle ergometer through an equation that considers individual's body mass and external work rate, but not pedaling rate (PR). We hypothesized that including PR in the ACSM equation would improve its V˙o2 prediction accuracy. Ten healthy male participants' (age 19-48 years) were recruited and their steady-state V˙o2 was recorded on a cycle ergometer for 16 combinations of external work rates (0, 50, 100, and 150 W) and PR (50, 70, 90, and 110 revolutions per minute). V˙o2 was calculated by means of a new equation, and by the ACSM equation for comparison. Kinematic data were collected by means of an infrared 3-D motion analysis system in order to explore the mechanical determinants of V˙o2. Including PR in the ACSM equation improved the accuracy for prediction of sub-maximal V˙o2 during exercise (mean bias 1.9 vs. 3.3 mL O2 kg(-1) min(-1)) but it did not affect the accuracy for prediction of maximal V˙o2 (P > 0.05). Confirming the validity of this new equation, the results were replicated for data reported in the literature in 51 participants. We conclude that PR is an important determinant of human V˙o2 during cycling exercise, and it should be considered when predicting oxygen consumption. © 2015 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.

  17. Determination of trace elements and screening of metalloproteins in human blood and tissues

    International Nuclear Information System (INIS)

    Prohaska, K.

    2003-02-01

    Sequential and simultaneous atomic spectrometric detection was applied for the determination of metals in human whole blood, blood fractions and joint tissues. For this purpose ICP-OES (inductively coupled plasma - optical emission spectrometry) and GFAAS (graphite furnace - atomic absorption spectrometry) were optimized. The nebulizing technique of the ICP-OES causes a high consumption of the sample (2.4 mL /min uptake rate). Using the simultaneous modus of measurement it is possible to determine up to 20 elements in the samples of interest. Using GFAAS the elements can be detected sequentially only, the method is more time consuming in comparison. For the direct sample injection into the graphite tube only 20 aeL are needed. Since 1 mL sample has to be filled in the sample vessel of the autosampler, up to six elements can be determined in this volume. The most important advantage of the simultaneous multi-element detection is the low sample consumption, which is essential for analysis of biological samples. Normally only small amounts of human blood or tissues can be collected, and the concentrations of analytes are usually very low. Therefore in many cases a sample preparation is of advantage, which enables a pre-concentration of the analyte. The sample digestion was optimized with respect to the possible pre-concentration of the analytes. Ashing of the freeze-dried blood enabled a six fold higher concentration in the measuring solution compared with wet digestion of blood. For the ICP-OES sample introduction system two different nebulizers were tested in the complex matrix of the digested blood samples. A Meinhard and the microconcentric nebulizer were compared according to their analytical performance. The matrix of the samples caused LODs three to five times higher than in the aqueous standards. The application of the Meinhard nebulizer enabled sufficiently low LODs in the matrix for some elements of interest (Ca, Cu, Fe, Mg, P, S, Zn in freeze-dried blood

  18. Trace Elements in Human Myocardial Infarction Determined by Neutron Activation Analysis

    International Nuclear Information System (INIS)

    Wester, P.O.

    1965-05-01

    By means of neutron activation analysis, injured and adjacent uninjured human heart tissue from 12 autopsy cases with myocardial infarction are investigated with respect to the concentration of 23 trace elements. The bulk elements K, Na and P are also determined. A recently developed ion-exchange technique, combined with subsequent y-spectrometry, is used. The following trace elements are determined: Ag, As, Au, Ba, Br, Ca, Cd, Ce, Co, Cr, Cs, Cu, Fe, Hg, La, Mo, Rb, Sb, Sc, Se, Sm, Zn and W. In the injured tissue compared to the uninjured, calculation on a wet weight basis showed a decrease in Co, Cs, K, Mo, P, Rb and Zn, and an increase in Br, Ca, Ce, La, Na, Sb and Sm. The differences in Ca, La, Mo, P and Zn are dependent on the age of the myocardial infarction, and the regression lines for these elements are given. The concentration of the trace elements in uninjured tissue from infarcted hearts is compared to the concentration of these elements in normal heart tissue, determined in a previous study. In the uninjured tissue from infarcted hearts a decrease is found in Cu and Mo, and an increase in As and Ce

  19. Trace Elements in Human Myocardial Infarction Determined by Neutron Activation Analysis

    Energy Technology Data Exchange (ETDEWEB)

    Wester, P O

    1965-05-15

    By means of neutron activation analysis, injured and adjacent uninjured human heart tissue from 12 autopsy cases with myocardial infarction are investigated with respect to the concentration of 23 trace elements. The bulk elements K, Na and P are also determined. A recently developed ion-exchange technique, combined with subsequent y-spectrometry, is used. The following trace elements are determined: Ag, As, Au, Ba, Br, Ca, Cd, Ce, Co, Cr, Cs, Cu, Fe, Hg, La, Mo, Rb, Sb, Sc, Se, Sm, Zn and W. In the injured tissue compared to the uninjured, calculation on a wet weight basis showed a decrease in Co, Cs, K, Mo, P, Rb and Zn, and an increase in Br, Ca, Ce, La, Na, Sb and Sm. The differences in Ca, La, Mo, P and Zn are dependent on the age of the myocardial infarction, and the regression lines for these elements are given. The concentration of the trace elements in uninjured tissue from infarcted hearts is compared to the concentration of these elements in normal heart tissue, determined in a previous study. In the uninjured tissue from infarcted hearts a decrease is found in Cu and Mo, and an increase in As and Ce.

  20. Body Site Is a More Determinant Factor than Human Population Diversity in the Healthy Skin Microbiome.

    Directory of Open Access Journals (Sweden)

    Guillermo I Perez Perez

    Full Text Available We studied skin microbiota present in three skin sites (forearm, axilla, scalp in men from six ethnic groups living in New York City.Samples were obtained at baseline and after four days following use of neutral soap and stopping regular hygiene products, including shampoos and deodorants. DNA was extracted using the MoBio Power Lyzer kit and 16S rRNA gene sequences determined on the IIlumina MiSeq platform, using QIIME for analysis.Our analysis confirmed skin swabbing as a useful method for sampling different areas of the skin because DNA concentrations and number of sequences obtained across subject libraries were similar. We confirmed that skin location was the main factor determining the composition of bacterial communities. Alpha diversity, expressed as number of species observed, was greater in arm than on scalp or axilla in all studied groups. We observed an unexpected increase in α-diversity on arm, with similar tendency on scalp, in the South Asian group after subjects stopped using their regular shampoos and deodorants. Significant differences at phylum and genus levels were observed between subjects of the different ethnic origins at all skin sites.We conclude that ethnicity and particular soap and shampoo practices are secondary factors compared to the ecological zone of the human body in determining cutaneous microbiota composition.

  1. Somatomedin A in human serum: determined by radioimmunoassay and radioreceptorassay; results and implications

    International Nuclear Information System (INIS)

    Hall, K.; Sara, V.

    1981-01-01

    The polypeptide hormones in the somatomedin family were discovered as three different biological activities of serum: insulin-like activity (ILA), sulphation factor activity (SFA) and multiplication stimulatory activity (MSA). Three different biological actions on three different target tissues have been used to identify the somatomedins during their purification. All the somatomedins contain ILA, SFA and MSA but their order of potency of biological action varies from tissue to tissue. It is apparent that the pattern of somatomedin levels determined throughout human life varies according to the technique used for their determination. Each assay has a unqiue order of crossreaction for the various somatomedins, and is sensitive to different forms present in blood. Thus the different patterns must reflect the predominance of specific somatomedins at certain stages of life. The heterogeneity of polypeptide hormones in the circulating blood is well recognized but its relation to age and development has yet to be fully explored. Methods for the estimation of the somatomedins must take account of such differences. In the radioimmunoassay field it is generally recognized that each type of antibody has a unique recognition pattern. This applies equally to the radioreceptorassay where the specificity of the receptor varies according to species, organ and age. Consequently, it is essential that age-related standards are established for each technique for the determination of somatomedins. (Auth.)

  2. Determination of genetic relatedness from low-coverage human genome sequences using pedigree simulations.

    Science.gov (United States)

    Martin, Michael D; Jay, Flora; Castellano, Sergi; Slatkin, Montgomery

    2017-08-01

    We develop and evaluate methods for inferring relatedness among individuals from low-coverage DNA sequences of their genomes, with particular emphasis on sequences obtained from fossil remains. We suggest the major factors complicating the determination of relatedness among ancient individuals are sequencing depth, the number of overlapping sites, the sequencing error rate and the presence of contamination from present-day genetic sources. We develop a theoretical model that facilitates the exploration of these factors and their relative effects, via measurement of pairwise genetic distances, without calling genotypes, and determine the power to infer relatedness under various scenarios of varying sequencing depth, present-day contamination and sequencing error. The model is validated by a simulation study as well as the analysis of aligned sequences from present-day human genomes. We then apply the method to the recently published genome sequences of ancient Europeans, developing a statistical treatment to determine confidence in assigned relatedness that is, in some cases, more precise than previously reported. As the majority of ancient specimens are from animals, this method would be applicable to investigate kinship in nonhuman remains. The developed software grups (Genetic Relatedness Using Pedigree Simulations) is implemented in Python and freely available. © 2017 John Wiley & Sons Ltd.

  3. Determination of tetrahydrophtalimide and 2-thiothiazolidine-4-carboxylic acid, urinary metabolites of the fungicide captan, in rats and humans

    NARCIS (Netherlands)

    van Welie, R.T.H.; van Duyn, P; Lamme, E K; Jäger, P; van Baar, B L; Vermeulen, N P

    1991-01-01

    Capillary gas chromatographic (GC) methods using sulphur and mass selective detection for the qualitative and quantitative determination of tetrahydrophtalimide (THPI) and 2-thiothiazolidine-4-carboxylic acid (TTCA), urinary metabolites of the fungicide captan in rat and humans, were developed.

  4. DETERMINANTS OF HEALTHCARE EXPENDITURE ON HUMAN CAPITAL AND ECONOMIC GROWTH IN BANGLADESH: A LONGITUDINAL DATA ANALYSIS FROM 1995-2010

    OpenAIRE

    SHONGKOUR ROY

    2014-01-01

    The objective of this study was to examine the determinants of healthcare expenditure in Bangladesh between 1995 and 2010 byapplying the World development indicator data. First, I developed an empirical model for longitudinal data analysis to determinant thehealthcare expenditure under the human capital and economic growth. Next, I explored the dynamic relationships among healthcareexpenditure, human capital and economic growth usingOLSmodel. The expenditure in private healthcare was 2.3% and...

  5. Determinants of human papillomavirus vaccine acceptability in Latin America and the Caribbean.

    Science.gov (United States)

    Winkler, Jennifer L; Wittet, Scott; Bartolini, Rosario M; Creed-Kanashiro, Hilary M; Lazcano-Ponce, Eduardo; Lewis-Bell, Karen; Lewis, Merle J; Penny, Mary E

    2008-08-19

    Prophylactic human papillomavirus (HPV) vaccines provide promise as a key component of future cervical cancer prevention programs in the Latin America and the Caribbean region. The successful introduction and acceptance of these vaccines will depend on a range of factors including awareness of cervical cancer as a problem, affordability of the vaccine, political will, competition with other vaccines, feasibility of vaccine delivery and acceptability of the vaccine among the range of groups who will influence uptake. While existing data about acceptability from Latin America and the Caribbean is scarce, it is clear that health policymakers, providers and the general public lack knowledge about HPV and cervical cancer. Furthermore, they would value more local epidemiologic data related to cervical cancer. Price is currently a major barrier to vaccine acceptability and a priority for advocacy. More research is required in Latin America and the Caribbean to determine what messages and strategies will work in these communities.

  6. Emotionally intelligent leadership as a key determinant of strategic and effective management of human capital

    Directory of Open Access Journals (Sweden)

    Helena Viskupičová

    2016-06-01

    Full Text Available The purpose of this paper is to outline the strategic importance of Emotional Intelligence (EI in contemporary approaches to management of Human resources (HR. The aim of this study is to provide a conceptual theoretical insight of the relationship between EI and leadership. The paper also reflects the conditions within the present business environment in Slovakia with the emphasis on leadership in the context of EI (based on the research conducted in this field, while answering the following question: To what extent do Slovak organizations incorporate EI measures into the hiring process for managerial positions? The answer to this question would consequently disclose whether organizations in Slovakia consider Emotional Intelligence as a factor determining effective performance of managers – leaders.

  7. Headspace Solid Phase Micro Extraction Gas Chromatographic Determination of Fenthion in Human Serum

    Directory of Open Access Journals (Sweden)

    Kyriaki Machera

    2008-05-01

    Full Text Available A simple and effective analytical procedure was developed for the determination of fenthion residues in human serum samples. The sample treatment was performed using the headspace solid-phase micro extraction with polyacrylate fiber, which has the advantage to require low amount of serum (1 mL without tedious pre-treatment. The quantification of fenthion was carried out by gas chromatography-mass spectrometry and the recoveries ranged from 79 to 104% at two spiking levels for 6 replicates. Detection and quantification limits were calculated as 1.51 and 4.54 ng/mL of serum respectively. Two fenthion metabolites − fenoxon and fenthion–sulfoxide − were also identified.

  8. Gas Chromatographic Mass Spectrometric Determination of Myo-inositol in Humans Utilizing a Deuterated Internal Standard

    DEFF Research Database (Denmark)

    Andersen, Jan Rud; Larsen, Elfinn; Harbo, Helge

    1982-01-01

    The isotopic dilution technique was used for determining the content of myo-inositol in human urine, plasma and haemolysed erythrocyte samples. A deuterated myo-inositol, synthesized from inosose-2 by base-catalysed exchange of hydrogens by deuterium, followed by reduction of the inosose with 2H2......, was added as internal standard to the samples at an early stage in the analytical procedure. After separation and derivatization to the hexa-acetate, the gas chromatographic mass spectrometric analysis was carried out. A 25 m fused silica capillary column coated with methyl silicone was used, and the ions...... selected for monitoring were m/z 210 and m/z 214, which are characteristic and abundant fragment ions from unlabelled and hexadeuterated myo-inositolhexa-acetate, respectively. Calibration curves from water, urine, plasma and haemolysed erythrocytes show parallel, linear responses in the ratio between...

  9. Determination of clarithromycin in human plasma by liquid chromatography-electrospray ionization tandem mass spectrometry.

    Science.gov (United States)

    Jiang, Yao; Wang, Jiang; Li, Hao; Wang, Yingwu; Gu, Jingkai

    2007-03-12

    A rapid and sensitive method has been developed for the determination of clarithromycin in human plasma with liquid chromatography-tandem mass spectrometry. Clarithromycin and the internal standard, telmisartan were precipitated from the matrix (50 microl) with 200 microl acetonitrile and separated by HPLC using formic acid:10 mM ammonium acetate:methanol (1:99:400, v/v/v) as the mobile phase. The assay based on detection by electrospray positive ionization mass spectrometry in the multiple-reaction monitoring mode was finished within 2.4 min. Linearity was over the concentration range 10-5000 ng/ml with a limit of detection of 0.50 ng/ml. Intra- and inter-day precision measured as relative standard deviation were bioequivalence study of two tablet formulations of clarithromycin.

  10. Application of PIGE to determine fluorine concentration in human teeth. Contribution to fluorosis study

    International Nuclear Information System (INIS)

    Salah, H.; Arab, N.

    2007-01-01

    Fluorosis, osteoselerosis, and systemic illness may result when tolerance levels of fluorine intake are exceeded. To avoid these abnormalities, we shall be aware of the mechanisms of their development. For this, quantitative results gathered from different parts of the world are necessary. In this paper, PIGE method is used to determine fluorine concentration in human teeth, collected from different regions of the Algerian Sahara. Both enamel and dentine regions were inspected. The external and internal mappings of the analyzed teeth have shown more variation in the localized fluorine concentration in dentine than in enamel, and the lowest level of fluorine concentration was observed in the frontier crown-root zone. The obtained results reveal relatively high fluorine concentration lying within the limit of fluorosis, according to the standardised grading dental fluorosis. (author)

  11. Determination of knowledge of Turkish midwifery students about human papilloma virus infection and its vaccines.

    Science.gov (United States)

    Genc, Rabia Ekti; Sarican, Emine Serap; Turgay, Ayse San; Icke, Sibel; Sari, Dilek; Saydam, Birsen Karaca

    2013-01-01

    Human papilloma virus (HPV) is one of the most common sexually transmitted agents and its infection is the most established cause of cervical cancer. Midwives play a key position in the implementation of cervical cancer. This descriptive study aimed to determine the level of knowledge concerning HPV and HPV vaccination among 268 midwifery students. Data were collected between November 15 and 30, 2011, through a self-reported questionnaire. The mean age of participants was 20.75 ± 1.60. Among all students, 44.4% had heard of HPV, while 40.4% had heard of HPV vaccinatiob. The relationship between the midwifery student knowledge on HPV and HPV vaccine and their current educational year was significant (p=0.001). In conclusion midwifery students have moderate level of knowledge about HPV and its vaccine and relevant information should be included in their teaching curriculum.

  12. Arsenic determination in water supplies for human consumption of the province of Cartago, Costa Rica

    International Nuclear Information System (INIS)

    Montero-Campos, Virginia; Quesada-Kimsey, Jaime; Ledezma-Espinoza, Aura; Sandoval-Mora, Jose A.

    2010-01-01

    Scientific knowledge about hydroarsenicism must be disclosed in Latin America. The presence of arsenic has been detected in waters of Costa Rica that have been used for human consumption, in areas of risk; specifically in the province of Cartago, in the cantons of Oreamuno, Central, Paraiso and Alvarado. A quantification of reduced form trivalent arsenic was performed with the methodology of the 7062 Environmental Protection Agency of the United States, through volatile metal hydride generation by flame atomic absorption. The analyzed samples have determined that maintaining the maximum extent permitted by current legislation Costa Rican, 10μg/L. Research areas have corresponded to areas of high risk for its volcanic nature. The vast water supply of the cantons under study, with areas of relative protection circling recharge areas of springs, has caused the population to consume water source underground, but surface direct influence, less influence of volcanic rocks that are found at greater depths. (author) [es

  13. Determination of tritium concentrations in humans before the development of a nuclear power plant in Turkey.

    Science.gov (United States)

    Dizman, Serdar; Yilmaz, Adnan; Keser, Recep

    2015-01-01

    The most widely used method to determine the level of tritium in humans is testing urine. Tritium concentrations in urine samples of 100 persons aged 18-66 years selected randomly from a pilot region in Turkey were analysed. The average activity concentration of urine samples was 4.66 ± 1.94 Bq L(-1) and the maximum activity concentration was 27.91 Bq L(-1). The minimum detectable activity was 2.38 Bq L(-1). The annual effective dose from tritium was also evaluated on the basis of the measurement results and reference values recommended by the International Commission on Radiological Protection. The effective doses for males and females were 4.56 and 3.54 nSv, respectively. These results were lower than the permissible annual effective dose for members of the public.

  14. [Simultaneous determination of eight kinds of conjunct bile acids in human bile by R-HPLC].

    Science.gov (United States)

    Dai, Z; Tan, G; Qian, K; Chen, X

    1997-01-01

    A method for the simultaneous determination of eight kinds of conjunct bile acids in human bile was developed by HPLC. They were separated on a YWG-C18 (3 microns) column at 30 degrees C, with methanol/water (65/35, V/V, pH3.0) as mobile phase, and detection wavelength at UV 210 nm. The linear ranges were 50-1,000 microns.ml-1, the recoveries were 91.2%-108.6%. The biles of 30 cases with cholelithiasis cholecystolithiasis and 20 cases without gallstone were detected by HPLC. The results showed that the constitution of bile acids was different between patients with cholelithiasis cholecystolithiasis and patients without gallstone.

  15. Reference measurement procedure for the determination of electrolytes in human blood via ICP-OES measurement

    Science.gov (United States)

    Grote-Koska, D.; Klauke, R.; Brand, K.; Schumann, G.

    2018-04-01

    The determination of electrolytes in human body fluids is one of the most frequently performed analyses in clinical routine laboratories. Metrological traceability of measurement results in patient samples is essential and requires the involvement of higher order reference measurement procedures wherever available. Here, the authors present the evaluation of a higher order reference system for the simultaneous determination of K+, Li+, Na+, Ca2+ and Mg2+ in blood serum and plasma. In the same order, the determined measurement performances were as follows: measurement ranges: 0.75 mmol l-1-75.0 mmol l-1, 0.05 mmol l-1-5.00 mmol l-1, 5 mmol l-1-200 mmol l-1, 0.4 mmol l-1-8.0 mmol l-1 and 0.1 mmol l-1-4.0 mmol l-1. Measurement imprecision: CVs were  ⩽1.1% for intra assay investigations and  ⩽1.8% for long term inter assay investigations for all measurands. Excellent accuracy was found testing certified Standard Reference Materials from NIST: SRM 909 (deviations from 0.0% to 1.1%) and SRM 956 (deviations from 0.0% to 1.5%). Intercomparisons with the German Metrology Institute (PTB) revealed differences from 0.1% to 0.8%. Matrix influences and carry over were not detectable. The expanded combined measurement uncertainties for the determination of the reference method values were estimated as  ⩾1.5% (k  =  2) for each measurand. The reference measurement procedure is accredited by the German accreditation body (DAkkS) in association with the German calibration service (DKD) according to ISO 17025 and ISO 15195. Services comprise the certification of calibrators, control materials and samples used in proficiency testing schemes.

  16. Determining The Factors Causing Human Error Deficiencies At A Public Utility Company

    Directory of Open Access Journals (Sweden)

    F. W. Badenhorst

    2004-11-01

    Full Text Available According to Neff (1977, as cited by Bergh (1995, the westernised culture considers work important for industrial mental health. Most individuals experience work positively, which creates a positive attitude. Should this positive attitude be inhibited, workers could lose concentration and become bored, potentially resulting in some form of human error. The aim of this research was to determine the factors responsible for human error events, which lead to power supply failures at Eskom power stations. Proposals were made for the reduction of these contributing factors towards improving plant performance. The target population was 700 panel operators in Eskom’s Power Generation Group. The results showed that factors leading to human error can be reduced or even eliminated. Opsomming Neff (1977 soos aangehaal deur Bergh (1995, skryf dat in die westerse kultuur werk belangrik vir bedryfsgeestesgesondheid is. Die meeste persone ervaar werk as positief, wat ’n positiewe gesindheid kweek. Indien hierdie positiewe gesindheid geïnhibeer word, kan dit lei tot ’n gebrek aan konsentrasie by die werkers. Werkers kan verveeld raak en dit kan weer lei tot menslike foute. Die doel van hierdie navorsing is om die faktore vas te stel wat tot menslike foute lei, en wat bydra tot onderbrekings in kragvoorsiening by Eskom kragstasies. Voorstelle is gemaak vir die vermindering van hierdie bydraende faktore ten einde die kragaanleg se prestasie te verbeter. Die teiken-populasie was 700 paneel-operateurs in die Kragopwekkingsgroep by Eskom. Die resultate dui daarop dat die faktore wat aanleiding gee tot menslike foute wel verminder, of geëlimineer kan word.

  17. Analytical determination of specific 4,4'-methylene diphenyl diisocyanate hemoglobin adducts in human blood.

    Science.gov (United States)

    Gries, Wolfgang; Leng, Gabriele

    2013-09-01

    4,4'-Methylene diphenyl diisocyanate (MDI) is one of the most important isocyanates in the industrial production of polyurethane and other MDI-based synthetics. Because of its high reactivity, it is known as a sensitizing agent, caused by protein adducts. Analysis of MDI is routinely done by determination of the nonspecific 4,4'-methylenedianiline as a marker for MDI exposure in urine and blood. Since several publications have reported specific adducts of MDI and albumin or hemoglobin, more information about their existence in humans is necessary. Specific adducts of MDI and hemoglobin were only reported in rats after high-dose MDI inhalation. The aim of this investigation was to detect the hemoglobin adduct 5-isopropyl-3-[4-(4-aminobenzyl)phenyl]hydantoin (ABP-Val-Hyd) in human blood for the first time. We found values up to 5.2 ng ABP-Val-Hyd/g globin (16 pmol/g) in blood samples of workers exposed to MDI. Because there was no information available about possible amounts of this specific MDI marker, the analytical method focused on optimal sensitivity and selectivity. Using gas chromatography-high-resolution mass spectrometry with negative chemical ionization, we achieved a detection limit of 0.02 ng ABP-Val-Hyd/g globin (0.062 pmol/g). The robustness of the method was confirmed by relative standard deviations between 3.0 and 9.8 %. Combined with a linear detection range up to 10 ng ABP-Val-Hyd/g globin (31 pmol/g), the enhanced precision parameter demonstrates that the method described is optimized for screening studies of the human population.

  18. Determination of the effectiveness of inactivation of human immunodeficiency virus by Pretoria pasteurization.

    Science.gov (United States)

    Jeffery, B S; Webber, L; Mokhondo, K R; Erasmus, D

    2001-12-01

    The risk of transmission of the human immunodeficiency virus (HIV) via breastfeeding is between 10 and 17 per cent. In resource-poor countries most HIV-infected women cannot afford to formula feed their infants and formula feeding is not desirable in areas of high infant mortality because of loss of the immunological benefits of breastmilk. A method has been devised by which HIV-infected women may express and pasteurize their breastmilk in a domestic setting using inexpensive apparatus and a simple technique. The method, Pretoria Pasteurization has been shown to be reliable under a wide range of conditions and maintains milk between 56 degrees and 62.5 degrees C for between 12 and 15 min. This study was devised to determine whether Pretoria Pasteurization effectively inactivates HIV in human milk. Samples of expressed breastmilk were obtained from a group of HIV-infected lactating women and a group of HIV-negative women. The samples of milk from the HIV-negative women were inoculated with high titres of cell-associated and cell-free HIV. Each sample was divided into a control portion and a study portion. The study portion underwent Pretoria Pasteurization. Control and pasteurized samples were inoculated into lymphocyte co-culture for a period of 35 days. All co-cultures were sampled weekly and analysed by serological and molecular methods for p24 antigen, cell-free HIV RNA and integrated DNA. Viral RNA was detected in the milk of 80 per cent amongst the known HIV-positive women. The mean serum viral load in the group of HIV positive women was 50728 copies/ml and the mean milk viral load was 422000 copies/ml. Evidence of viral replication was shown in 11 of the control specimens. There was no evidence of viral replication in any of the study specimens which had undergone Pretoria Pasteurization. It was concluded that Pretoria Pasteurization effectively inactivates HIV in human milk.

  19. Determination of molybdenum in human urine by electrothermal atomization atomic absorption spectrometry

    International Nuclear Information System (INIS)

    Pita Calvo, C.; Bermejo Barrera, P.; Bermejo Barrera, A.

    1995-01-01

    Various matrix modifiers were investigated for the determination of molybdenum in human urine samples by electrothermal atomization atomic absorption spectrometry. Methods with nitric acid, barium difluoride, magnesium nitrate, palladium-magnesium nitrate and palladium-hydroxylamine hydrochloride were studied by introducing the urine samples directly into the graphite furnace with 0.3% Triton X-100. The charring and atomization curves, the amount of modifier and the calibration and addition graphs were studied in all instances. The precision, accuracy and chemical interferences of the methods were also investigated. The matrix interferences have been removed with the modifiers barium difluoride, palladium-magnesium nitrate and palladium-hydroxylamine hydrochloride. The limits of detection and quantification were 0.2 and 0.7 μg l -1 , respectively, for these modifiers. The characteristic masses were 14.1, 18.0 and 14.9 pg of Mo for palladium-magnesium nitrate, palladium-hydroxylamine hydrochloride and barium difluoride, respectively. The method with palladium-magnesium nitrate has been applied to the study of the amount of molybdenum in human urine samples. The molybdenum levels found lie between 4.8-205.6 μg l -1

  20. Simultaneous determination of ramipril, ramiprilat and telmisartan in human plasma using liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Gupta, V K; Jain, Rajeev; Lukram, Ojitkumar; Agarwal, Shilpi; Dwivedi, Ashish

    2011-01-15

    A rapid and sensitive liquid chromatography tandem mass spectrometry method has been developed and validated for the simultaneous determination of ramipril, ramiprilat and telmisartan in human plasma. The solid-phase extraction technique was used for the extraction of ramipril, ramiprilat and telmisartan from human plasma. Trandolaprilat and hydrochlorothiazide were used as the internal standards (ISs). Chromatography was performed on a Hypurity C18, 5 μm, 50 mm × 4.6mm column, with the mobile phase consisting of ammonium acetate and acetonitrile (in a 20:80 ratio), followed by detection using mass spectrometry. The method involves a simple reversed isocratic chromatography condition and mass spectrometry detection, which enables detection at sub-nanogram levels. The method was validated and the lower limit of quantification for ramipril, ramiprilat and telmisartan was found to be 0.1 ng mL(-1), 0.1 ng mL(-1) and 2 ng mL(-1), respectively. The mean recovery for ramipril, ramiprilat and telmisartan ranged from 90.1 to 104.1%. This method increased the sensitivity and selectivity; resulting in high-throughput analysis of ramipril, ramiprilat and telmisartan using two different ISs in a single experiment for bioequivalence studies, with a chromatographic run time of 1.5 min only. Copyright © 2010 Elsevier B.V. All rights reserved.

  1. Development and validation of HPLC analytical method for quantitative determination of metronidazole in human plasma

    International Nuclear Information System (INIS)

    Safdar, K.A.; Shyum, S.B.; Usman, S.

    2016-01-01

    The objective of the present study was to develop a simple, rapid and sensitive reversed-phase high performance liquid chromatographic (RP-HPLC) analytical method with UV detection system for the quantitative determination of metronidazole in human plasma. The chromatographic separation was performed by using C18 RP column (250mm X 4.6mm, 5 meu m) as stationary phase and 0.01M potassium dihydrogen phosphate buffered at pH 3.0 and acetonitrile (83:17, v/v) as mobile phase at flow rate of 1.0 ml/min. The UV detection was carried out at 320nm. The method was validated as per the US FDA guideline for bioanalytical method validation and was found to be selective without interferences from mobile phase components, impurities and biological matrix. The method found to be linear over the concentration range of 0.2812 meu g/ml to 18.0 meu g/ml (r2 = 0.9987) with adequate level of accuracy and precision. The samples were found to be stable under various recommended laboratory and storage conditions. Therefore, the method can be used with adequate level of confidence and assurance for bioavailability, bioequivalence and other pharmacokinetic studies of metronidazole in human. (author)

  2. Determination of trace uranium in human hair by nuclear track detection technique

    International Nuclear Information System (INIS)

    Chung, Yong Sam; Moon, Jong Hwa; En, Zinaida; Cho, Seung Yeon; Kang, Sang Hoon; Lee, Jae Ki

    2001-01-01

    The aim of this study is to describe a usefulness of nuclear analytical technique in assessing and comparing the concentration levels through the analysis of uranium using human hair sample in the field of environment. A fission track detection technique was applied to determine the uranium concentration in human hair. Hair samples were collected from two groups of people - a) workers not dealing with uranium directly, and b) workers possibly contaminated with uranium. The concentration of 235 U for the first group varied from <1 to 39 ng/g and the second group can be estimated up to the level of μg/g. Radiographs of heavy-duty work samples contained high dense 'hot spots' along a single hair. After washing in acetone and distilled water, external contamination was not totally removed. Insoluble uranium compounds were not completely washed out. The (n, f)-radiography technique, having high sensitivity, and capable of getting information on uranium content at each point of a single hair, is an excellent tool for environmental monitoring

  3. Determination of uranium in human hair by acid digestion and FIAS-ICPMS

    International Nuclear Information System (INIS)

    Gonnen, R.; Kol, R.; Laichter, Y.; Marcus, P.; Lorber, A.; Karpas, Z.; Halicz, L.

    2000-01-01

    The content of heavy metals in human hair may serve as an indicator of occupational or environmental exposure of metal compounds. However, before such exposure can be determined, the level of the element in a 'normal' population must be established. The concentration of uranium in human hair was measured by flow injection - inductively coupled plasma mass spectrometry (FIAS-ICPMS) after acid digestion of the hair samples. All hair samples were rinsed in order to remove external contamination prior to the digestion in a 2:1 solution of concentrated nitric acid and 30% hydrogen peroxide. The limit of detection of the method, for a 50 mg hair sample, was 0.015 μg/g, mainly due to the presence of impurities in the hydrogen peroxide. The range of uranium concentration in the initial test group was found to be 0.01-0.18 μg/g. The mean and median values of the entire study population were 0.062 and 0.050 μg U/g hair, respectively. Differences between the following sub-populations: male and female, smokers and non-smokers and people below and above 45 years of age were examined. The only statistically significant difference was found in the latter group (p=0.03). (author)

  4. Carbon dots based immunosorbent assay for the determination of GFAP in human serum

    Science.gov (United States)

    Ma, Yunsu; Xu, Guanhong; Wei, Fangdi; Cen, Yao; Song, Yueyue; Ma, Yujie; Xu, Xiaoman; Shi, Menglan; Sohail, Muhammad; Hu, Qin

    2018-04-01

    Glial fibrillary acidic protein (GFAP) is expressed in the central nervous system and the level of GFAP normally rises with brain injury and astroglial tumors. So, serum GFAP is used as a marker for diagnosing various types of brain damage and astroglial tumors. In this study, a new sensor based on carbon dots (CDs) linked with antibodies to specifically detect GFAP in human serum was developed. Anti-GFAP (Ab1) linked with protein A/G agarose resin (PA/G) as a capture antibody (PA/G-Ab1) and anti-GFAP (Ab2) labeled with CDs as a detection antibody (CDs-Ab2) were prepared firstly. Then the CD-linked antibody immunosorbent assay (CLAISA) method was constructed based on the sandwich conjunction reaction among PA/G-Ab1, GFAP, and CDs-Ab2. CLAISA, using the fluorescence of PA/G-Ab1-GFAP-Ab2-CDs as the direct signal, enabled the proposed immunosensor to detect GFAP sensitively with a linear range of 0.10-8.00 ng ml-1 and a detection limit of 25 pg ml-1. This method was applied to the determination of GFAP in human serum by the standard addition method, and the results showed high accuracy and precision. Considering the easy synthetic process and excellent performance of CLAISA, this method has great potential to be used to monitor GFAP in the clinic.

  5. Sex determination of human mandible using metrical parameters by computed tomography: A prospective radiographic short study

    Directory of Open Access Journals (Sweden)

    Basavaraj N Kallalli

    2016-01-01

    Full Text Available Introduction: Sex determination of unidentified human remains is very important in forensic medicine, medicolegal cases, and forensic anthropology. The mandible is the largest and hardest facial bone that commonly resists postmortem damage and forms an important source of personal identification. Additional studies have demonstrated the applicability of facial reconstruction using three-dimensional computed tomography scan (3D-CT for the purpose of individual identification. Aim: To determine the sex of human mandible using metrical parameters by CT. Materials and Methods: The study included thirty subjects (15 males and 15 females, with age group ranging between 10 and 60 years obtained from the outpatient department of Oral Medicine and Radiology, Narsinhbhai Patel Dental College and Hospital. CT scan was performed on all the subjects, and the data obtained were reconstructed for 3D viewing. After obtaining 3D-CT scan, a total of seven mandibular measurements, i.e., gonial angle (G-angle, ramus length (Ramus-L, minimum ramus breadth and gonion-gnathion length (G-G-L, bigonial breadth, bicondylar breadth (BIC-Br, and coronoid length (CO-L were measured; collected data were analyzed using SPSS statistical analysis program by Student's t-test. Results: The result of the study showed that out of seven parameters, G-angle, Ramus-L, G-G-L, BIC-Br, and CO-L showed a significant statistical difference (P < 0.05, with overall accuracy of 86% for males and 82% for females. Conclusion: Personal identification using mandible by conventional methods has already been proved but with variable efficacies. Advanced imaging modalities can aid in personal identification with much higher accuracy than conventional methods.

  6. Simple method for the determination of rosiglitazone in human plasma using a commercially available internal standard.

    Science.gov (United States)

    Mamidi, Rao N V S; Benjamin, Biju; Ramesh, Mullangi; Srinivas, Nuggehally R

    2003-09-01

    To the best of our knowledge, bioanalytical methods to determine rosiglitazone in human plasma reported in literature use internal standards that are not commercially available. Our purpose was to develop a simple method for the determination of rosiglitazone in plasma employing a commercially available internal standard (IS). After the addition of celecoxib (IS), plasma (0.25 mL) samples were extracted into ethyl acetate. The residue after evaporation of the organic layer was dissolved in 750 microL of mobile phase and 50 microL was injected on to HPLC. The separation was achieved using a Hichrom KR 100, 250 x 4.6 mm C(18) with a mobile phase composition potassium dihydrogen phosphate buffer (0.01 m, pH 6.5):acetonitrile:methanol (40:50:10, v/v/v). The flow-rate of the mobile phase was set at 1 mL/min. The column eluate was monitored by fluorescence detector set at an excitation wavelength of 247 nm and emission wavelength of 367 nm. Linear relationships (r(2) > 0.99) were observed between the peak area ratio rosiglitazone to IS vs rosiglitazone concentrations across the concentration range 5-1000 ng/mL. The intra-run precision (%RSD) and accuracy (%Dev) in the measurement of rosiglitazone were 80% for both rosiglitazone and IS from human plasma. The lower limit of quantitation of the assay was 5 ng/mL. In summary, the methodology for rosiglitazone measurement in plasma was simple, sensitive and employed a commercially available IS. Copyright 2003 John Wiley & Sons, Ltd.

  7. Two different spectrofluorimetric methods for simultaneous determination of gemfibrozil and rosiglitazone in human plasma.

    Science.gov (United States)

    El-Din, Mohie M K Sharaf; Attia, Khalid A M; Nassar, Mohamed W I; Kaddah, Mohamed M Y

    2010-10-15

    Two accurate, reliable, and highly sensitive spectrofluorimetric methods were developed for simultaneous determination of binary mixture gemfibrozil and rosiglitazone in human plasma without prior separation steps. The first method is based on synchronous fluorescence spectrometry using double scans. At Δλ=27nm, gemfibrozil yields detectable signal that is independent of the presence of rosiglitazone. Similarly, at Δλ=120nm the signal of rosiglitazone is not influenced by the presence of gemfibrozil. Signals at two wavelengths, 301 (Δλ=27nm) and 368nm (Δλ=120nm) vary linearly with gemfibrozil and rosiglitazone concentrations over the range 100-700ngmL(-1) (for gemfibrozil) and 20-140ngmL(-1) (for rosiglitazone), respectively. The limits of detection (LOD) were 2.3 and 2.72ngmL(-1) for gemfibrozil and rosiglitazone, respectively. The second method is based on the technique of simultaneous equations (Vierodt's method), in which 258nm was selected as the excitation wavelength. Two equations are constructed based on the fact that at ( λ(EM)₂=302 nm of gemfibrozil) and (λ(EM)₂=369 nm of rosiglitazone) the fluorescence of the mixture is the sum of the individual fluorescence of gemfibrozil and rosiglitazone. The limits of detection (LOD) were 28.1 and 23.63ngmL(-1) for gemfibrozil and rosiglitazone, respectively. The proposed methods were successfully applied for the determination of the two compounds in synthetic mixtures and in human plasma with a good recovery. Copyright © 2010 Elsevier B.V. All rights reserved.

  8. The viral transcription group determines the HLA class I cellular immune response against human respiratory syncytial virus.

    Science.gov (United States)

    Johnstone, Carolina; Lorente, Elena; Barriga, Alejandro; Barnea, Eilon; Infantes, Susana; Lemonnier, François A; David, Chella S; Admon, Arie; López, Daniel

    2015-04-01

    The cytotoxic T-lymphocyte-mediated killing of virus-infected cells requires previous recognition of short viral antigenic peptides bound to human leukocyte antigen class I molecules that are exposed on the surface of infected cells. The cytotoxic T-lymphocyte response is critical for the clearance of human respiratory syncytial virus infection. In this study, naturally processed viral human leukocyte antigen class I ligands were identified with mass spectrometry analysis of complex human leukocyte antigen-bound peptide pools isolated from large amounts of human respiratory syncytial virus-infected cells. Acute antiviral T-cell response characterization showed that viral transcription determines both the immunoprevalence and immunodominance of the human leukocyte antigen class I response to human respiratory syncytial virus. These findings have clear implications for antiviral vaccine design. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  9. The Viral Transcription Group Determines the HLA Class I Cellular Immune Response Against Human Respiratory Syncytial Virus*

    Science.gov (United States)

    Johnstone, Carolina; Lorente, Elena; Barriga, Alejandro; Barnea, Eilon; Infantes, Susana; Lemonnier, François A.; David, Chella S.; Admon, Arie; López, Daniel

    2015-01-01

    The cytotoxic T-lymphocyte-mediated killing of virus-infected cells requires previous recognition of short viral antigenic peptides bound to human leukocyte antigen class I molecules that are exposed on the surface of infected cells. The cytotoxic T-lymphocyte response is critical for the clearance of human respiratory syncytial virus infection. In this study, naturally processed viral human leukocyte antigen class I ligands were identified with mass spectrometry analysis of complex human leukocyte antigen-bound peptide pools isolated from large amounts of human respiratory syncytial virus-infected cells. Acute antiviral T-cell response characterization showed that viral transcription determines both the immunoprevalence and immunodominance of the human leukocyte antigen class I response to human respiratory syncytial virus. These findings have clear implications for antiviral vaccine design. PMID:25635267

  10. "Determination of mycophenolic acid in human plasma by high-performance liquid chromatography"

    Directory of Open Access Journals (Sweden)

    "Mehdi Ahadi Barzoki

    2005-05-01

    Full Text Available A simple, sensitive and reproducible HPLC method is presented for determination of mycophenolic acid(MPA in human plasma. Samples were prepared after precipitation of the plasma protein by addition of acetonitrile and naproxen was used as internal standard (I.S.. Separation was performed by reversedphase HPLC, using a Hamilton PRP-C18 Column, 51% acetonitrile and 49% potassium phosphate buffer (20 mM at pH 3.0 as mobile phase, flow rate of 1.0 ml/min, and UV detection at 215 nm. MPA and I.S. had retention times of 7.5 and 11.35 min, respectively. The method showed an acceptable linearity in the range of 0.1µg/ml-40µg/ml with r2 of .9992. The concentration of 0.1µg/ml was determined as quantification limit. Mean absolute recovery was 94.8%. The mean intra- and inter-day reproducibility of method was 4.6 and 11.4% respectively.

  11. A research agenda for helminth diseases of humans: social ecology, environmental determinants, and health systems.

    Directory of Open Access Journals (Sweden)

    Andrea Gazzinelli

    Full Text Available In this paper, the Disease Reference Group on Helminth Infections (DRG4, established in 2009 by the Special Programme for Research and Training in Tropical Diseases (TDR, with the mandate to review helminthiases research and identify research priorities and gaps, focuses on the environmental, social, behavioural, and political determinants of human helminth infections and outlines a research and development agenda for the socioeconomic and health systems research required for the development of sustainable control programmes. Using Stockols' social-ecological approach, we describe the role of various social (poverty, policy, stigma, culture, and migration and environmental determinants (the home environment, water resources development, and climate change in the perpetuation of helminthic diseases, as well as their impact as contextual factors on health promotion interventions through both the regular and community-based health systems. We examine these interactions in regard to community participation, intersectoral collaboration, gender, and possibilities for upscaling helminthic disease control and elimination programmes within the context of integrated and interdisciplinary approaches. The research agenda summarises major gaps that need to be addressed.

  12. Lack of close relationship between three strains of human rhinoviruses as determined by their RNA sequences.

    Science.gov (United States)

    Yin, F H; Lonberg-Holm, K; Chan, S P

    1973-07-01

    The possible genomic homologies between three serotypes of human rhinoviruses (HRV 1A, HRV 2, and HRV 14) were investigated. First we confirmed that these viruses were unrelated by the criterion of the absence of common antigenic determinants on the surfaces of the native virions, as detected by cross-neutralization of complementfixation. RNA-RNA hybridization was then examined with purified, highly radioactive, double-stranded, replicative-form RNA and excess single-stranded virion RNA. Single-stranded RNA showed 100% homology with the minus strand from the replicative-form RNA of the same type of virus. HRV 1A, HRV 2, and HRV 14 showed low intertypic homologies; these were not significantly greater than those found between the rhinoviruses and polivirus, which were used as a negative control. The immunological relationship and the RNA homology between HRV 1A and HRV 1B were also examined by the above techniques. It was confirmed that HRV 1A and HRV 1B share some surface determinants and it was also found that HRV 1B RNA shares 70% homology with HRV 1A RNA.

  13. Structural determinants of phenotypic diversity and replication rate of human prions.

    Directory of Open Access Journals (Sweden)

    Jiri G Safar

    2015-04-01

    Full Text Available The infectious pathogen responsible for prion diseases is the misfolded, aggregated form of the prion protein, PrPSc. In contrast to recent progress in studies of laboratory rodent-adapted prions, current understanding of the molecular basis of human prion diseases and, especially, their vast phenotypic diversity is very limited. Here, we have purified proteinase resistant PrPSc aggregates from two major phenotypes of sporadic Creutzfeldt-Jakob disease (sCJD, determined their conformational stability and replication tempo in vitro, as well as characterized structural organization using recently emerged approaches based on hydrogen/deuterium (H/D exchange coupled with mass spectrometry. Our data clearly demonstrate that these phenotypically distant prions differ in a major way with regard to their structural organization, both at the level of the polypeptide backbone (as indicated by backbone amide H/D exchange data as well as the quaternary packing arrangements (as indicated by H/D exchange kinetics for histidine side chains. Furthermore, these data indicate that, in contrast to previous observations on yeast and some murine prion strains, the replication rate of sCJD prions is primarily determined not by conformational stability but by specific structural features that control the growth rate of prion protein aggregates.

  14. Characterisation of monoclonal antibodies for human luteinising hormone, and mapping of antigenic determinants on the hormone

    International Nuclear Information System (INIS)

    Soos, M.; Siddle, K.

    1983-01-01

    Twelve mouse monoclonal antibodies for human luteinising hormone were produced. The affinities varied from 4 X 10 7 to 1 X 10 10 l/mol. The specificity of each antibody was assessed by determining the relative reactivities with luteinising hormone, thyroid stimulating hormone, follicle stimulating hormone and chorionic gonadotrophin. Six antibodies bound to the α-subunit as shown by similar reactivity with all hormones, and the remainder to the β-subunit as shown by specificity for luteinising hormone. This latter group of antibodies cross-reacted only weakly with thyroid stimulating hormone (approximately 10%) and follicle stimulating hormone (approximately 3%). Three of these antibodies also showed low reactivity towards chorionic gonadotrophin (<10%), though the others did not (80-300%). The ability of different antibodies to bind simultaneously to luteinising hormone was examined and it was shown that several distinct antigenic determinants existed on both subunits. The characterisation of monoclonal binding sites is discussed in relation to the use of antibodies in two-site immunoradiometric assays. (Auth.)

  15. Sensitive method for the determination of nitrated polycyclic aromatic hydrocarbons in the human diet

    Energy Technology Data Exchange (ETDEWEB)

    Siegmund, B.; Weiss, R.; Pfannhauser, W. [Graz Univ. of Technology (Austria). Inst. of Food Chemistry

    2003-01-01

    The presence of nitrated polycyclic aromatic hydrocarbons (nitro-PAHs) as contaminants in foods is nowadays a legitimate cause of concern, as they are reported to be strong direct-acting mutagens and carcinogens that may pose a risk to human health. Nevertheless, their concentrations in foods are in the ultra-trace region. In this study, we present a very sensitive analytical method for the determination of this compound class in complex food matrices. Special emphasis was put on sample extraction and clean up. Recoveries of 75% could be obtained for 1- and 2-nitronaphthalene; for all other compounds investigated the recoveries were {>=}94%. The determination of the compounds was performed by gas chromatography-mass spectrometry. The results show that the use of negative chemical ionization (NCI) increases the sensitivity by one order of magnitude in comparison to electron impact ionization (EI) (limits of detection 0.07-0.25 {mu}g kg{sup -1} for EI and 0.01-0.02 {mu}g kg{sup -1} for NCI). The influence of the sample matrix on the sensitivity of the method is demonstrated with extracts of a sample from a duplicate diet study. Due to the lack of a certified reference material the suitability of the method is demonstrated with pumpkin seed oil that was spiked with nitro-PAHs at two different concentration levels. (orig.)

  16. Simultaneous determination of rifabutin and human serum albumin in pharmaceutical formulations by capillary electrophoresis.

    Science.gov (United States)

    Ermolenko, Yu; Anshakova, A; Osipova, N; Kamentsev, M; Maksimenko, O; Balabanyan, V; Gelperina, S

    Capillary zone electrophoresis (CZE) was used for determination of rifabutin (RFB), an anti-tuberculosis antibiotic drug, in various pharmaceutical formulations. Apart from that, simultaneous determination of RFB and human serum albumin (HSA) was performed. Electrophoretic behaviour of RFB was examined at various pH levels. CE conditions: a quartz capillary tube (internal diameter 75mm, effective length 50cm, total length 60cm), the capillary temperature was 25°С, the voltage applied to the capillary tube was +20kV, the UV detection wavelength was 214nm, hydrodynamic injection of the sample was performed at 30mbar for 5s, tetraborate buffer solution (0.01М, рН9.2). The obtained results are characterized by high efficiency (number of theoretical plates up to 260,000) and sufficient sensitivity (LOQ starting from 0.02μg/ml for RFB). The obtained data are in good accord with both HPLC results (for RFB) and spectrophotometry (for HSA). Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Spectrophotometric determination of mefenamic acid excreted as free drug in urine of human beings

    International Nuclear Information System (INIS)

    Naseer, M.M.; Nawaz, R.; Shafique, M.; Rehman, R.

    2007-01-01

    Urinary excretion of free mefenamic acid was investigated in 16 healthy human volunteers, eight males and eight females, following the oral administration of 500 mg tablet of mefenamic acid. Urine samples were collected at pre-determined schedule and drug concentration was determined by spectrophotometric method. The total recovery of free mefenamic acid was 1.526 +- 0.128 and 1.193 +- 0.112% in male and female volunteers respectively. The average +- S.E values for diuresis, pH and rate of excretion of mefenamic acid was 0.0160 +- 0.004 mL/min./kg of body weight, 6.22 +- 0.167, 0.077 +- 0.016 micro g min/sup -1/kg/sup -1/in male while 0.0084 +- 0.0023mL min/sup -1/kg-1 of body weight, 6.35 +- 0.164, 0.054 +- 0.008 micro g min/sup -1/kg/sup -1/respectively in female volunteers. The results obtained are different from the earlier studies due to variability in dose, gender variation, fluctuation in urine pH, environmental conditions and nutritional ingredients. (author)

  18. Microdose clinical trial: quantitative determination of nicardipine and prediction of metabolites in human plasma.

    Science.gov (United States)

    Yamane, Naoe; Takami, Tomonori; Tozuka, Zenzaburo; Sugiyama, Yuichi; Yamazaki, Akira; Kumagai, Yuji

    2009-01-01

    A sample treatment procedure and high-sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for quantitative determination of nicardipine in human plasma were developed for a microdose clinical trial with nicardipine, a non-radioisotope labeled drug. The calibration curve was linear in the range of 1-500 pg/mL using 1 mL of plasma. Analytical method validation for the clinical dose, for which the calibration curve was linear in the range of 0.2-100 ng/mL using 20 microL of plasma, was also conducted. Each method was successfully applied to making determinations in plasma using LC/MS/MS after administration of a microdose (100 microg) and clinical dose (20 mg) to each of six healthy volunteers. We tested new approaches in the search for metabolites in plasma after microdosing. In vitro metabolites of nicardipine were characterized using linear ion trap-fourier transform ion cyclotron resonance mass spectrometry (LIT-FTICRMS) and the nine metabolites predicted to be in plasma were analyzed using LC/MS/MS. There is a strong possibility that analysis of metabolites by LC/MS/MS may advance to utilization in microdose clinical trials with non-radioisotope labeled drugs.

  19. Human haptoglobin phenotypes and concentration determination by nanogold-enhanced electrochemical impedance spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Cheng, Tsai-Mu; Lee, Tzu-Cheng; Tseng, Shin-Hua; Chu, Hsueh-Liang; Chang, Chia-Ching [Department of Biological Science and Technology, National Chiao Tung University, Hsinchu 30050, Taiwan (China); Pan, Ju-Pin, E-mail: ccchang01@faculty.nctu.edu.tw [Division of Cardiology, Department of Medicine, Taipei Veterans General Hospital, Taipei 11217, Taiwan (China)

    2011-06-17

    Haptoglobin (Hp) is an acute phase protein that binds free hemoglobin (Hb), preventing Hb-induced oxidative damage in the vascular system. There are three phenotypes in human Hp, whose heterogeneous polymorphic structures and varying concentrations in plasma have been attributed to the cause of diseases and outcome of clinical treatments. Different phenotypes of Hp may be composed of the same subunits but different copy numbers, rendering their determination difficult by a single procedure. In this study, we have developed a simple, fast, reliable and sensitive method, using label-free nanogold-modified bioprobes coupled with self-development electrochemical impedance spectroscopy (EIS). By this method, probe surface charge transfer resistance is detected. The relative charge transfer resistance ratios for Hp 1-1, Hp 2-1 and Hp 2-2 were characterized. We were able to determine protein size difference within 3 nm, and the linear region of the calibration curve for Hp levels in the range of 90 pg ml{sup -1} and 90 {mu}g ml{sup -1} ({approx}1 fM to 1 pM). We surmise that similar approaches can be used to investigate protein polymorphism and altered protein-protein interaction associated with diseases.

  20. Determination of natural in vivo noble-gas concentrations in human blood.

    Directory of Open Access Journals (Sweden)

    Yama Tomonaga

    Full Text Available Although the naturally occurring atmospheric noble gases He, Ne, Ar, Kr, and Xe possess great potential as tracers for studying gas exchange in living beings, no direct analytical technique exists for simultaneously determining the absolute concentrations of these noble gases in body fluids in vivo. In this study, using human blood as an example, the absolute concentrations of all stable atmospheric noble gases were measured simultaneously by combining and adapting two analytical methods recently developed for geochemical research purposes. The partition coefficients determined between blood and air, and between blood plasma and red blood cells, agree with values from the literature. While the noble-gas concentrations in the plasma agree rather well with the expected solubility equilibrium concentrations for air-saturated water, the red blood cells are characterized by a distinct supersaturation pattern, in which the gas excess increases in proportion to the atomic mass of the noble-gas species, indicating adsorption on to the red blood cells. This study shows that the absolute concentrations of noble gases in body fluids can be easily measured using geochemical techniques that rely only on standard materials and equipment, and for which the underlying concepts are already well established in the field of noble-gas geochemistry.

  1. Fluorine determination in human healthy and carious teeth using the PIGE technique

    International Nuclear Information System (INIS)

    Carvalho, M.L.; Karydas, A.G.; Casaca, C.; Zarkadas, Ch.; Paradellis, Th.; Kokkoris, M.; Nsouli, B.; Cunha, A.S.

    2001-01-01

    The purpose of this study is to determine and compare the fluorine concentration in human teeth from two different populations, living in the Portuguese quite isolated islands of Acores: S. Miguel and Terceira. Both populations have similar dietary habits, similar occupational activities, mostly rural, and the age of both populations is more or less the same, around 40 years. No chronic diseases were registered in any of the donors. The two groups are exposed to different levels of fluorine in drinking water. Terceira island has moderate fluorine concentration levels (1-2 μg g -1 ) while S. Miguel island is known for the high fluorine concentration levels in its water (>3 μg g -1 ), especially in one area known as Furnas. Thirty-three teeth, 17 healthy and 16 carious without restoration (14 incisors and canines, 7 premolars and 12 molars), were collected and analyzed for the determination of fluorine concentration in the dentine region, using the nuclear reaction 19 F(p,αγ) 16 O. The teeth were cross-sectioned along the vertical plane and polished, in order to obtain a smooth and plane surface of about 1 mm thickness. In this work an association between caries prevalence and fluorine content of drinking water is discussed and the variation of fluorine concentration among different types of teeth (canines and incisors, premolars, molars) and physical state (carious and non-carious) is examined

  2. Ex vivo irradiation of human blood to determine DNA damage using molecular techniques

    International Nuclear Information System (INIS)

    Montes, Angel; Agapito, Juan

    2014-01-01

    Biological dosimetry is the assessment of absorbed dose in individuals exposed to ionizing radiation from blood samples based on the radiation induced damage in cellular DNA. The aim of this study was to determine the damage in the DNA through the assessment of an experimental ex vivo assay using irradiated samples of human blood cells. For this purpose, blood samples were irradiated at low doses (<100 mGy) considering the following parameters: blood volume (3mL), temperature (37 °C) and incubation time (0.5, 2, 4, 8 and 24 h). Dose values were: 0, 12.5, 25 and 50 mGy using Cesium -137 gamma rays at 662 keV and a dose rate of 38.46 mGy/h. The qualitative damage in the genomic DNA was determined using agarose gel electrophoresis and polymerase chain reaction (PCR) for the p53 gene in a sequence of 133 pb of exon 7, related to the protein that acts in the cell repair process. The results of the qualitative analysis showed no degradation of genomic DNA; also an increase in the DNA concentration was observed up to the fourth hour of incubation, finding maximum values for all doses in the two samples. As a conclusion, the effects of ionizing radiation at doses used in this experiment do not generate a detectable damage, by means of molecular techniques such as those used in the present study. (authors).

  3. Determination of linsidomine in human plasma by tandem LC-MS with ESI.

    Science.gov (United States)

    Sutherland, F C; de Jager, A D; Swart, K J; Hundt, H K; Scanes, T; Hundt, A F

    2000-04-01

    A sensitive method for the determination of linsidomine in plasma was developed, using high-performance liquid chromatographic (HPLC) separation with tandem mass spectrometric detection. Linsidomine was derivatised with propyl chloroformate and extracted with tert-butyl methyl ether/1,2-dichloroethane (55:45, v/v), back-extracted into HCl (0.01 M) followed by alkalinisation and back-extraction into ether; the final ether extract evaporated, reconstituted in mobile phase and then separated on a Phenomenex Luna C18 (2) 5 micron 2.1 x 150 mm column with a mobile phase consisting of methanol water formic acid (98/100%) (400:600:0.05, v/v/v) at a flow-rate of 0.4 ml min(-1). Detection was achieved by a Finnigan MAT mass spectrometer (LCQ) at unit resolution in the selected reaction monitoring (SRM) mode monitoring the transition of the protonated molecular ion m/z 257.0 to the product ion m/z 86.0. The mean recovery for linsidomine was 51% with a lower limit of quantification of 0.70 ng/ml using 1 ml plasma for extraction. This LC-MS/MS method for the determination of linsidomine in human plasma allows for better specificity and a higher sample throughput than the traditional LC-UV methods. It also demonstrates the profound effect that the composition of acidic modifiers and matrix constituents can have on the electrospray ionisation (ESI) of the analyte.

  4. Electrochemistry of raloxifene on glassy carbon electrode and its determination in pharmaceutical formulations and human plasma.

    Science.gov (United States)

    Bagheri, Akbar; Hosseini, Hadi

    2012-12-01

    The electrochemical behavior of raloxifene (RLX) on the surface of a glassy carbon electrode (GCE) has been studied by cyclic voltammetry (CV). The CV studies were performed in various supporting electrolytes, wide range of potential scan rates, and pHs. The results showed an adsorption-controlled and quasi-reversible process for the electrochemical reaction of RLX, and a probable redox mechanism was suggested. Under the optimum conditions, differential pulse voltammetry (DPV) was applied for quantitative determination of the RLX in pharmaceutical formulations. The DPV measurements showed that the anodic peak current of the RLX was linear to its concentration in the range of 0.2-50.0μM with a detection limit of 0.0750μM, relative standard deviation (RSD %) below 3.0%, and a good sensitivity. The proposed method was successfully applied for determination of the RLX in pharmaceutical and human plasma samples with a good selectivity and suitable recovery. Copyright © 2012 Elsevier B.V. All rights reserved.

  5. Standardization of the radioimmunoassay technique for the determination of human gastrin and its clinical application

    International Nuclear Information System (INIS)

    Peig Ginabredra, M.G.

    1989-01-01

    It was developed and standardized a system of radioimmunoassay for the determination of gastrin, employing synthetic human gastrin for radioiodination and preparation of standard as well as specific antibody raised rabbits. The hormone was labeled with 125 I by the Cloramine T techique and purified by anion exchange chromatography in QAE-Sephadex A-25, being determined its specific activity. The tracer thus obtained was submitted to analysis of purity by poliacrilamide gel eletrophoresis and precipitation of proteins by trichloroacetic acid. Its stability evaluated according to the time of storage, being its purity and adequation for the use in radioimmunoassay also compared to a tracer obtained from a commercial diagnosis kit. The assays were performed by incubation of radioiodinated gastrin, standard gastrin prepared in plasma free from this hormone (from zero to 500 pmol/l) or samples to be assayed with the antiserum for 4 days at 4 0 C. The separation between the free gastrin and the gastrin bound to the antibody was carried out by adsorption of the free hormone to the charcoal, whose ideal concentration was previously determined. Plasma free from gastrin was obtained from time-expired blood bank plasma submitted to extraction with charcoal. When performed the quality control, this radioimmunoassay was shown specific, accurate, precise and sensitive, allowing the performance of valid assays. Its validation was even confirmed by clear discrimination not only of the gastrin concentration in subjects with very low levels (gastrectomized) and extremely high levels (Zollinger-Ellison syndrome) as well as gastrin concentrations in subjects with other diseases, such as Chagas disease, pernicious anemia and chronic renal failure. (author) [pt

  6. Flow-cytometric determination of high-density-lipoprotein binding sites on human leukocytes

    International Nuclear Information System (INIS)

    Schmitz, G.; Wulf, G.; Bruening, T.A.; Assmann, G.

    1987-01-01

    In this method, leukocytes were isolated from 6 mL of EDTA-blood by density-gradient centrifugation and subsequently incubated with rhodamine isothiocyanate (RITC)-conjugated high-density lipoproteins (HDL). The receptor-bound conjugate particles were determined by fluorescent flow cytometry and compared with 125 I-labeled HDL binding data for the same cells. Human granulocytes express the highest number of HDL binding sites (9.4 x 10(4)/cell), followed by monocytes (7.3 x 10(4)/cell) and lymphocytes (4.0 x 10(4)/cell). Compared with conventional analysis of binding of 125 I-labeled HDL in tissue-culture dishes, the present determination revealed significantly lower values for nonspecific binding. In competition studies, the conjugate competes for the same binding sites as 125 I-labeled HDL. With the use of tetranitromethane-treated HDL3, which fails to compete for the HDL receptor sites while nonspecific binding is not affected, we could clearly distinguish between 37 degrees C surface binding and specific 37 degrees C uptake of RITC-HDL3, confirming that the HDL receptor leads bound HDL particles into an intracellular pathway rather than acting as a docking type of receptor. Patients with familial dysbetalipoproteinemia showed a significantly higher number of HDL binding sites in the granulocyte population but normal in lymphocytes and monocytes, indicating increased uptake of cholesterol-containing lipoproteins. In patients with familial hypercholesterolemia, HDL binding was increased in all three cell types, indicating increased cholesterol uptake and increased cholesterol synthesis. The present method allows rapid determination of HDL binding sites in leukocytes from patients with various forms of hyper- and dyslipoproteinemias

  7. Spectrofluorimetric determination of sertraline in dosage forms and human plasma through derivatization with 9-fluorenylmethyl chloroformate

    Directory of Open Access Journals (Sweden)

    Belal Fathalla

    2011-10-01

    Full Text Available Abstract Background Sertraline is primarily used to treat major depression in adult outpatients as well as obsessive-compulsive, panic and social anxiety disorders in both adults and children. A survey of the literature reveals that most of the reported methods are either insufficiently sensitive or tedious and require highly sophisticated and dedicated instrumentation. The proposed method is considered to be specific for determination of SER in presence of its metabolite (deaminated form. Results A sensitive, simple and specific spectrofluorimetric method was developed for the determination of sertraline (SER in pharmaceutical formulations and biological fluids. The method is based on its reaction with 9-fluorenylmethyl chloroformate (FMOC-Cl in borate buffer of pH 8.0 to yield a highly fluorescent derivative peaking at 315 nm after excitation at 265 nm. The different experimental parameters affecting the development and stability of the reaction product were carefully studied and optimized. The fluorescence concentration plot was rectilinear over the range of 0.05-1.0 μg mL-1 with a lower detection limit of 5.34 × 10-3 μg mL-1 and limit of quantitation of 0.016 μg mL-1. Conclusions The proposed method was successfully applied to the analysis of commercial tablets and the results obtained were in good agreement with those obtained using the reference method. Furthermore, the method was applied for the determination of SER in spiked and real human plasma. The mean % recovery (n = 3 was 94.33 ± 1.53 and 92.00 ± 2.65, respectively. A proposal of the reaction pathway was postulated.

  8. Topographic antigenic determinants recognized by monoclonal antibodies on human choriogonadotropin beta-subunit

    International Nuclear Information System (INIS)

    Bidart, J.M.; Troalen, F.; Salesse, R.; Bousfield, G.R.; Bohuon, C.J.; Bellet, D.H.

    1987-01-01

    We describe a first attempt to study the antibody-combining sites recognized by monoclonal antibodies raised against the beta-subunit of human choriogonadotropin (hCG). Two groups of antibodies were first defined by their ability to recognize only the free beta-subunit or the free and combined subunit. Antibodies FBT-11 and FBT-11-L bind only to hCG beta-subunit but not to hCG, whereas antibodies FBT-10 and D1E8 bind to both the beta-subunit and the hormone. In both cases, the antigenic determinants were localized to the core of the protein (residues 1-112), indicating the weak immunogenicity of the specific carboxyl-terminal extension of hCG-beta. Nine synthetic peptides spanning different regions of hCG-beta and lutropin-beta were assessed for their capacity to inhibit antibody binding. A synthetic peptide inclusive of the NH2-terminal region (residues 1-7) of the hCG beta-subunit was found to inhibit binding to the radiolabeled subunit of a monoclonal antibody specific for free hCG-beta (FBT-11). Further delineation of the antigenic site recognized by this antibody provided evidence for the involvement of fragment 82-92. Moreover, monoclonal antibody FBT-11 inhibited the recombination of hCG-beta to hCG-alpha, indicating that its antigenic determinant might be located nearby or in the hCG-beta portion interacting with the alpha-subunit. Binding of monoclonal antibody FBT-10, corresponding to the second antigenic determinant, was weakly inhibited by fragment 82-105 and did not impair the recombination of the hCG beta-subunit to the hCG alpha-subunit. Its combining site appeared to be located in a region of the intact native choriogonadotropin present at the surface of the hormone-receptor complex

  9. Simultaneous Estimation and Validation for Determination of Lamivudine and Zidovudine in Human Plasma By LCMS/MS Method

    Directory of Open Access Journals (Sweden)

    C. Parthiban

    2012-01-01

    Full Text Available A high performance liquid chromatographic method with mass detection was developed for determination of Lamivudine and Zidovudine in human plasma. Lamivudine and Zidovudine were extracted from an aliquot of human plasma using Solid Phase Extraction technique, and then injected into a liquid chromatograph equipped with a tandem mass spectrometry detector; quantitation was done by peak area ratio method. A weighted (1/Conc2 linear regression was performed to determine the concentration of analyte. This method demonstrates acceptable performance and is suitable for the determination of Lamivudine and Zidovudine in human plasma over the range of 25.291 to 4046.621 ng/mL and 23.357 to 4057.l4lng/mL respectively using Solid Phase Extraction Procedure. As all the values obtained were within the limits as specified, the method has been successfully used to analyse the human plasma containing Lamivudine and Zidovudine with good recoveries and proved to be robust.

  10. Determining

    Directory of Open Access Journals (Sweden)

    Bahram Andarzian

    2015-06-01

    Full Text Available Wheat production in the south of Khuzestan, Iran is constrained by heat stress for late sowing dates. For optimization of yield, sowing at the appropriate time to fit the cultivar maturity length and growing season is critical. Crop models could be used to determine optimum sowing window for a locality. The objectives of this study were to evaluate the Cropping System Model (CSM-CERES-Wheat for its ability to simulate growth, development, grain yield of wheat in the tropical regions of Iran, and to study the impact of different sowing dates on wheat performance. The genetic coefficients of cultivar Chamran were calibrated for the CSM-CERES-Wheat model and crop model performance was evaluated with experimental data. Wheat cultivar Chamran was sown on different dates, ranging from 5 November to 9 January during 5 years of field experiments that were conducted in the Khuzestan province, Iran, under full and deficit irrigation conditions. The model was run for 8 sowing dates starting on 25 October and repeated every 10 days until 5 January using long-term historical weather data from the Ahvaz, Behbehan, Dezful and Izeh locations. The seasonal analysis program of DSSAT was used to determine the optimum sowing window for different locations as well. Evaluation with the experimental data showed that performance of the model was reasonable as indicated by fairly accurate simulation of crop phenology, biomass accumulation and grain yield against measured data. The normalized RMSE were 3%, 2%, 11.8%, and 3.4% for anthesis date, maturity date, grain yield and biomass, respectively. Optimum sowing window was different among locations. It was opened and closed on 5 November and 5 December for Ahvaz; 5 November and 15 December for Behbehan and Dezful;and 1 November and 15 December for Izeh, respectively. CERES-Wheat model could be used as a tool to evaluate the effect of sowing date on wheat performance in Khuzestan conditions. Further model evaluations

  11. Alpha-CaMKII plays a critical role in determining the aggressive behavior of human osteosarcoma.

    Science.gov (United States)

    Daft, Paul G; Yuan, Kaiyu; Warram, Jason M; Klein, Michael J; Siegal, Gene P; Zayzafoon, Majd

    2013-04-01

    Osteosarcoma is among the most frequently occurring primary bone tumors, primarily affecting adolescents and young adults. Despite improvements in osteosarcoma treatment, more specific molecular targets are needed as potential therapeutic options. One target of interest is α-Ca(2+)/calmodulin-dependent protein kinase II (α-CaMKII), a ubiquitous mediator of Ca(2+)-linked signaling, which has been shown to regulate tumor cell proliferation and differentiation. Here, we investigate the role of α-CaMKII in the growth and tumorigenicity of human osteosarcoma. We show that α-CaMKII is highly expressed in primary osteosarcoma tissue derived from 114 patients, and is expressed in varying levels in different human osteosarcoma (OS) cell lines [MG-63, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)/HOS, and 143B). To examine whether α-CaMKII regulates osteosarcoma tumorigenic properties, we genetically inhibited α-CaMKII in two osteosarcoma cell lines using two different α-CaMKII shRNAs delivered by lentiviral vectors and overexpressed α-CaMKII by retrovirus. The genetic deletion of α-CaMKII by short hairpin RNA (shRNA) in MG-63 and 143B cells resulted in decreased proliferation (50% and 41%), migration (22% and 25%), and invasion (95% and 90%), respectively. The overexpression of α-CaMKII in HOS cells resulted in increased proliferation (240%), migration (640%), and invasion (10,000%). Furthermore, α-CaMKII deletion in MG-63 cells significantly reduced tumor burden in vivo (65%), whereas α-CaMKII overexpression resulted in tumor formation in a previously nontumor forming osteosarcoma cell line (HOS). Our results suggest that α-CaMKII plays a critical role in determining the aggressive phenotype of osteosarcoma, and its inhibition could be an attractive therapeutic target to combat this devastating adolescent disease. ©2013 AACR.

  12. Determination of N-methylsuccinimide and 2-hydroxy-N-methylsuccinimide in human urine and plasma.

    Science.gov (United States)

    Jönsson, B A; Akesson, B

    1997-12-19

    A method for determination of N-methylsuccinimide (MSI) and 2-hydroxy-N-methylsuccinimide (2-HMSI) in human urine and of MSI in human plasma was developed. MSI and 2-HMSI are metabolites of the widely used organic solvent N-methyl-2-pyrrolidone (NMP). MSI and 2-HMSI were purified from urine and plasma by C8 solid-phase extraction and analysed by gas chromatography-mass spectrometry in the negative-ion chemical ionisation mode. The intra-day precisions in urine were 2-6% for MSI (50 and 400 ng/ml) and 3-5% for 2-HMSI (1000 and 8000 ng/ml). For MSI in plasma it was 2% (60 and 1200 ng/ml). The between-day precisions in urine were 3-4% for MSI (100 and 1000 ng/ml) and 2-4% for 2-HMSI (10,000 and 18,000 ng/ml) and 3-4% for MSI in plasma (100 and 900 ng/ml). The recoveries from urine were 109-117% for MSI (50 and 400 ng/ml) and 81-89% for 2-HMSI (1000 and 8000 ng/ml). The recovery of MSI from plasma was 91-101% (50 and 500 ng/ml). The detection limits for MSI were 3 ng/ml in urine and 1 ng/ml in plasma and that of 2-HMSI in urine was 200 ng/ml. The method is applicable for analysis of urine and plasma samples from workers exposed to NMP.

  13. Simultaneous determination of psychotropic drugs in human urine by capillary electrophoresis with electrochemiluminescence detection

    Energy Technology Data Exchange (ETDEWEB)

    Li Jianguo [Key Laboratory of Analytical Chemistry for Life Science (Education Ministry of China), School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093 (China); School of Chemistry and Chemical Engineering, Suzhou University, Suzhou 215006 (China); Zhao Fengjuan [Key Laboratory of Analytical Chemistry for Life Science (Education Ministry of China), School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093 (China); Ju Huangxian [Key Laboratory of Analytical Chemistry for Life Science (Education Ministry of China), School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093 (China)]. E-mail: hxju@nju.edu.cn

    2006-08-04

    Amitriptyline, doxepin and chlorpromazine are often used as psychotropic drugs in treatment of the various mental diseases, and are also partly excreted by kidney. This work developed a simple, selective and sensitive method for their simultaneous monitoring in human urine using capillary electrophoresis coupled with electrochemiluminescence (ECL) detection based on end-column ECL reaction of tris-(2,2'-bipyridyl)ruthenium(II) with aliphatic tertiary amino moieties. Acetone was used as an additive to the running buffer to obtain their absolute separation. Under optimized conditions the proposed method displayed a linear range from 5.0 to 800 ng mL{sup -1} for the three drugs with the correlation coefficients more than 0.995 (n = 8). Their limits of detection were 0.8 ng mL{sup -1} (3.6 fg), 1.0 ng mL{sup -1} (4.5 fg) and 1.5 ng mL{sup -1} (6.8 fg) at a signal to noise ratio of 3, respectively. The relative standard deviations for five determinations of 20 ng mL{sup -1} amitriptyline, doxepin and chlorpromazine were 1.7%, 4.2% and 3.6%, respectively. For practical application an extract step with 90:10 heptane/ethyl acetate (v/v) was performed to eliminate the influence of ionic strength in sample. The recoveries of amitriptyline, doxepin and chlorpromazine at different levels in human urine were between 83% and 93%, which showed that the method was valuable in clinical and biochemical laboratories for monitoring amitriptyline, doxepin and chlorpromazine.

  14. Sensitive spectrophotometric determination of metoclopramide hydrochloride in dosage forms and spiked human urine using vanillin

    Directory of Open Access Journals (Sweden)

    O. Zenita Devi

    2016-09-01

    Full Text Available A new spectrophotometric method which is simple, sensitive, selective and rapid is described for the determination of metoclopramide hydrochloride (MCP in bulk drug and in dosage forms using vanillin as the chromogenic agent. The method is based on the condensation reaction between primary aromatic amine group present in MCP with aromatic aldehyde, vanillin to produce an intense yellow colored product. The resulting Schiff’s base shows an absorption maximum at 410 nm and the reaction product is stable for more than one day. The reaction was carried out in acetic acid and perchloric acid medium. Beer’s law was obeyed in the concentration range 1.5–15.0 μg ml−1 MCP with a molar absorptivity of 1.89 × 104 l mol−1 cm−1. The limit of detection (LOD and limit of quantification (LOQ were found to be 0.51 and 1.55 μg ml−1, respectively. The method was statistically evaluated by calculating percent relative error (% RE for accuracy and percent relative standard deviation (% RSD for precision, and was applied successfully to the determination of MCP in tablets, in injection and also in spiked human urine. No interference was observed from common additives found in pharmaceutical preparations. The results obtained by the proposed method were validated statistically by comparing the results with those of the reference method by applying the Student’s t-test and F-test. The accuracy and reliability of the method were further ascertained by performing recovery tests via standard-addition technique.

  15. A novel bioassay for the activity determination of therapeutic human brain natriuretic peptide (BNP.

    Directory of Open Access Journals (Sweden)

    Lei Yu

    Full Text Available BACKGROUND: Recombinant human brain natriuretic peptide (rhBNP is an important peptide-based therapeutic drug indicated for the treatment of acute heart failure. Accurate determination of the potency of therapeutic rhBNP is crucial for the safety and efficacy of the drug. The current bioassay involves use of rabbit aortic strips, with experiments being complicated and time-consuming and markedly variable in results. Animal-less methods with better precision and accuracy should be explored. We have therefore developed an alternative cell-based assay, which relies on the ability of BNP to induce cGMP production in HEK293 cells expressing BNP receptor guanylyl cyclase-A. METHODOLOGY/PRINCIPAL FINDINGS: An alternative assay based on the measurement of BNP-induced cGMP production was developed. Specifically, the bioassay employs cells engineered to express BNP receptor guanylyl cyclase-A (GCA. Upon rhBNP stimulation, the levels of the second messager cGMP in these cells drastically increased and subsequently secreted into culture supernatants. The quantity of cGMP, which corresponds to the rhBNP activity, was determined using a competitive ELISA developed by us. Compared with the traditional assay, the novel cell-based assay demonstrated better reproducibility and precision. CONCLUSION/SIGNIFICANCE: The optimized cell-based assay is much simpler, more rapid and precise compared with the traditional assay using animal tissues. To our knowledge, this is the first report on a novel and viable alternative assay for rhBNP potency analysis.

  16. Differences in gene expression of human xylosyltransferases and determination of acceptor specificities for various proteoglycans

    Energy Technology Data Exchange (ETDEWEB)

    Roch, Christina; Kuhn, Joachim; Kleesiek, Knut [Institut fuer Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum NRW, Universitaetsklinik der Ruhr-Universitaet Bochum, 32545 Bad Oeynhausen (Germany); Goetting, Christian, E-mail: cgoetting@hdz-nrw.de [Institut fuer Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum NRW, Universitaetsklinik der Ruhr-Universitaet Bochum, 32545 Bad Oeynhausen (Germany)

    2010-01-01

    The xylosyltransferase (XT) isoforms XT-I and XT-II initiate the posttranslational glycosaminoglycan (GAG) synthesis. Here, we determined the relative expression of both isoforms in 33 human cell lines. The majority of tested cell lines showed dominant XYLT2 gene expression, while only in 23132/87, JAR, NCI-H510A and THP-1 was the XT-I mRNA expression higher. Nearly equal expression levels were detected in six cell lines. Additionally, to shed light on putative differences in acceptor specificities the acceptor properties of potential acceptor sequences were determined. Peptides were expressed as glutathione-S-transferase fusion proteins containing putative or known GAG attachment sites of in vivo proteoglycans. Kinetic analysis showed that K{sub m} and V{sub max} values for XT-I mediated xylosylation were slightly higher than those for XT-II, and that XT-I showed a lesser stringency concerning the acceptor sequence. Mutagenesis of the bikunin peptide sequence in the G-S-G attachment site and flanking regions generated potential acceptor molecules. Here, mutations on the N-terminal side and the attachment site were found to be more susceptible to a loss of acceptor function than mutations in the C-terminus. Altogether the known consensus sequence a-a-a-a-G-S-G-a-a/G-a ('a' representing Asp or Glu) for XT-I mediated xylosylation could be approved and additionally extended to apply to XT-II as well.

  17. Determination of lipoic acid in human urine by capillary zone electrophoresis.

    Science.gov (United States)

    Kubalczyk, Paweł; Głowacki, Rafał

    2017-07-01

    Fast, simple, and accurate CE method enabling determination of lipoic acid (LA) in human urine has been developed and validated. LA is a disulfide-containing natural compound absorbed from the organism's diet. Due to powerful antioxidant activity, LA has been used for prevention and treatment of various diseases and disorders, e.g. cardiovascular diseases, neurodegenerative disorders, and cancer. The proposed analytical procedure consists of liquid-liquid sample extraction, reduction of LA with tris(2-carboxyethyl)phosphine, derivatization with 1-benzyl-2-chloropyridinium bromide (BCPB) followed by field amplified sample injection stacking, capillary zone electrophoresis separation, and ultraviolet-absorbance detection of LA-BCPB derivative at 322 nm. Effective baseline electrophoretic separation was achieved within 6 min under the separation voltage of 20 kV (∼80 μA) using a standard fused-silica capillary (effective length 51.5 cm, 75 μm id) and BGE consisted of 0.05 mol/L borate buffer adjusted to pH 9. The experimentally determined limit of detection for LA in urine was 1.2 μmol/L. The calibration curve obtained for LA in urine showed linearity in the range 2.5-80 μmol/L, with R 2 0.9998. The relative standard deviation of the points of the calibration curve was lower than 10%. The analytical procedure was successfully applied to analysis of real urine samples from seven healthy volunteers who received single 100 mg dose of LA. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Elasticity-based determination of isovolumetric phases in the human heart

    Directory of Open Access Journals (Sweden)

    Braun Jürgen

    2010-10-01

    Full Text Available Abstract Background/Motivation To directly determine isovolumetric cardiac time intervals by magnetic resonance elastography (MRE using the magnitude of the complex signal for deducing morphological information combined with the phase of the complex signal for tension-relaxation measurements. Methods Thirty-five healthy volunteers and 11 patients with relaxation abnormalities were subjected to transthoracic wave stimulation using vibrations of approximately 25 Hz. A k-space-segmented, ECG-gated gradient-recalled echo steady-state sequence with a 500-Hz bipolar motion-encoding gradient was used for acquiring a series of 360 complex images of a short-axis view of the heart at a frame rate of less than 5.2 ms. Magnitude images were employed for measuring the cross-sectional area of the left ventricle, while phase images were used for analyzing the amplitudes of the externally induced waves. The delay between the decrease in amplitude and onset of ventricular contraction was determined in all subjects and assigned to the time of isovolumetric tension. Conversely, the delay between the increase in wave amplitude and ventricular dilatation was used for measuring the time of isovolumetric elasticity relaxation. Results Wave amplitudes decreased during systole and increased during diastole. The variation in wave amplitude occurred ahead of morphological changes. In healthy volunteers the time of isovolumetric elasticity relaxation was 75 ± 31 ms, which is significantly shorter than the time of isovolumetric tension of 136 ± 36 ms (P n = 11 isovolumetric elasticity relaxation was significantly prolonged, with 133 ± 57 ms (P P = 0.053. Conclusion The complex MRE signal conveys complementary information on cardiac morphology and elasticity, which can be combined for directly measuring isovolumetric tension and elasticity relaxation in the human heart.

  19. (99m) Tc-labelled human serum albumin cannot replace (125) I-labelled human serum albumin to determine plasma volume in patients with liver disease

    DEFF Research Database (Denmark)

    Henriksen, Ulrik Lütken; Henriksen, Jens H; Bendtsen, Flemming

    2013-01-01

    Summary Background and aims Determination of plasma volume (PV) is important in several clinical situations. Thus, patients with liver disease often have augmented PV as part of their sodium–water retention. This study was undertaken to compare PV determination by two indicators: technetium......-labelled human serum albumin (99mTc-HSA) and iodine-labelled human serum albumin (125I-HSA), as the former may have advantages at repeated measurements and the latter is the classical gold standard. Study population and methods In 88 patients, (64 with liver disease, mainly cirrhosis, and 24 patients without...... In all patients, a close correlation was present between PV determined by the two indicators (r = 0·89, Pdetermined with 99mTc-HSA exceeded PV determined with 125I-HSA by 367 ml (5·2 ml kg...

  20. LC/MS Method for the Determination of Stable Isotope Labeled Promethazine in Human Plasma

    Science.gov (United States)

    Zuwei, Wang; Boyd, Jason; Berens, Kurt L.; Putcha, Lakshmi

    2004-01-01

    Promethazine (PMZ) is taken by astronauts orally (PO), intramuscularly (IM) or rectally (PR) for space motion sickness. LC/MS method was developed with off-line solid phase extraction to measure plasma concentrations of PMZ given as stable isotope-labeled (SIL) formulations by the three different routes of administration simultaneously. Samples (0.5ml) were loaded on to Waters Oasis HLB co-polymer cartridges and eluted with 1.0 mL methanol. HPLC separation of the eluted sample was performed using an Agilent Zorbax SB-CN column (50 x 2.1 mm) at a flow rate of 0.2 mL/min for 6 min. Acetonitrile/ ammonium acetate (30 mM) in water (3:2, v/v), pH 5.6 plus or minus 0.1, was used as the mobile phase for separation. Concentrations of PMZ, PMZ-d4 and PMZ-d7 and chlorpromazine (internal standard) were determined using a Micromass ZMD single quadrupole mass spectrometer with Electrospray Ionization (ESI). ESI mass spectra were acquired in positive ion mode with selected ion monitoring of [M+ H]dot plus. The method is rapid, reproducible and the assay specific parameters are listed in a table. A novel, sensitive and specific method for the measurement of PMZ and SIL PMZ in human plasma is reported.

  1. Determination of levofloxacin in human serum using liquid chromatography tandem mass spectrometry

    Directory of Open Access Journals (Sweden)

    Samiksha Ghimire

    2018-01-01

    Full Text Available A rapid liquid chromatography tandem-mass spectrometry method was developed for the determination of levofloxacin and its metabolite (desmethyl-levofloxacin in human serum. Sample preparation was done using protein precipitation technique. Our method had a run time of 2.5 min and retention times of 1.6 min for all analytes. The standard curves were linear within the concentration range of 0.10 to 5.00 mg/L for levofloxacin and 0.10 to 4.99 mg/L for desmethyl- levofloxacin; a correlation coefficient (R2 of 0.999 and 0.998 respectively. The lower limit of quantification for both analytes was 0.10 mg/L. Within-day precision ranged from 1.4% and 2.4% for levofloxacin, 1.5% to 5% for desmethyl-levofloxacin and between-day precision ranged from 3.6% to 4.1% for levofloxacin and 0.0% to 3.3% for desmethyl-levofloxacin; whereas, accuracy ranged from 0.1% to 12.7% for levofloxacin and 0.2% to 15.6% for desmethyl-levofloxacin. This method could be a useful asset for routine therapeutic drug monitoring of levofloxacin in multi-drug resistant tuberculosis patients.

  2. Determination of flomoxef in human plasma by liquid chromatography/electrospray ionization tandem mass spectrometry.

    Science.gov (United States)

    Kravtsova, Oxana Yu; Paramonov, Sergey A; Vasilevich, Natalya I; Kazyulkin, Denis N; Vlasova, Ekaterina; Engsig, Michael

    2013-12-01

    A specific, sensitive, rapid and reproducible method for the determination of flomoxef in human plasma using high-performance liquid chromatography-tandem mass spectrometry was developed and validated. Flomoxef was detected using an electrospay ionization method operated in negative-ion mode. Chromatographic separation was performed in gradient elution mode on a Luna® C18(2) column (3 μM, 20 × 4.0 mm) at a flow rate of 1 mL/min and runtime 3.5 min. The mobile phase consisted of acetonitrile and water containing 0.1% formic acid as additive. Extraction of flomoxef from plasma and precipitation of plasma proteins was performed with acetonitrile with an absolute recovery of 86.4 ± 1.6%. The calibration curve was linear with a correlation coefficient of 0.999 over the concentration range 10-5000 ng/mL and the lower limit of quantification was 10 ng/mL. The intra- and inter-day precisions were flomoxef revealed that it could be successfully analyzed at 4 ºС over 24 h, but it was unstable in solutions at room temperature during short-term storage (4 h) and several freeze-thaw cycles. Copyright © 2013 John Wiley & Sons, Ltd.

  3. DNA-binding determinants promoting NHEJ by human Polμ.

    Science.gov (United States)

    Martin, Maria Jose; Juarez, Raquel; Blanco, Luis

    2012-12-01

    Non-homologous end-joining (NHEJ), the preferred pathway to repair double-strand breaks (DSBs) in higher eukaryotes, relies on a collection of molecular tools to process the broken ends, including specific DNA polymerases. Among them, Polµ is unique as it can catalyze DNA synthesis upon connection of two non-complementary ends. Here, we demonstrate that this capacity is intrinsic to Polµ, not conferred by other NHEJ factors. To understand the molecular determinants of its specific function in NHEJ, the interaction of human Polµ with DNA has been directly visualized by electromobility shift assay and footprinting assays. Stable interaction with a DNA gap requires the presence of a recessive 5'-P, thus orienting the catalytic domain for primer and nucleotide binding. Accordingly, recognition of the 5'-P is crucial to align the two DNA substrates of the NHEJ reaction. Site-directed mutagenesis demonstrates the relevance of three specific residues (Lys(249), Arg(253) and Arg(416)) in stabilizing the primer strand during end synapsis, allowing a range of microhomology-induced distortions beneficial for NHEJ. Moreover, our results suggest that the Polµ BRCT domain, thought to be exclusively involved in interaction with NHEJ core factors, has a direct role in binding the DNA region neighbor to the 5'-P, thus boosting Polµ-mediated NHEJ reactions.

  4. Fractionation of human serum lipoproteins and simultaneous enzymatic determination of cholesterol and triglycerides

    International Nuclear Information System (INIS)

    Qureshi, Rashid Nazir; Kok, Wim Th.; Schoenmakers, Peter J.

    2009-01-01

    A method based on Asymmetric Flow Field-Flow Fractionation (AF4) was developed to separate different types of lipoproteins from human serum. The emphasis in the method optimization was on the possibilities to characterize the largest lipoprotein fractions (LDL and VLDL), which is usually not possible with the size-exclusion chromatography methods applied in routine analysis. Different channel geometries and flow programs were tested and compared. The use of a short fractionation channel was shown to give less sample dilution at the same fractionation power compared to a conventional, long channel. Different size selectivities were obtained with an exponential decay and a linear cross flow program. The ratio of the UV absorption signal to the light scattering signal was used to validate the relation between retention time and size of the fractionated particles. An experimental setup was developed for the simultaneous determination of the cholesterol and triglycerides distribution over the lipoprotein fractions, based on enzymatic reactions followed by UV detection at 500 nm. Coiled and knitted PTFE tubing reactors were compared. An improved peak sharpness and sensitivity were observed with the knitted tubing reactor. After optimization of the experimental conditions a satisfactory linearity and precision (2-3% rsd for cholesterol and 5-6% rsd for triglycerides) were obtained. Finally, serum samples, a pooled sample from healthy volunteers and samples of sepsis patients, were analyzed with the method developed. Lipoprotein fractionation and cholesterol and triglyceride distributions could be correlated with the clinical background of the samples.

  5. Determination of critical epitope of PcMab-47 against human podocalyxin

    Directory of Open Access Journals (Sweden)

    Shunsuke Itai

    2018-07-01

    Full Text Available Podocalyxin (PODXL is a type I transmembrane protein, which is highly glycosylated. PODXL is expressed in some types of human cancer tissues including oral, breast, and lung cancer tissues and may promote tumor growth, invasion, and metastasis. We previously produced PcMab-47, a novel anti-PODXL monoclonal antibody (mAb which reacts with endogenous PODXL-expressing cancer cell lines and normal cells independently of glycosylation in Western blot, flow cytometry, and immunohistochemical analysis. In this study, we used enzyme-linked immunosorbent assay (ELISA, flow cytometry, and immunohistochemical analysis to determine the epitope of PcMab-47. The minimum epitope of PcMab-47 was found to be Asp207, His208, Leu209, and Met210. A blocking peptide containing this minimum epitope completely neutralized PcMab-47 reaction against oral cancer cells by flow cytometry and immunohistochemical analysis. These findings could lead to the production of more functional anti-PODXL mAbs, which are advantageous for antitumor activities.

  6. Determination of critical epitope of PcMab-47 against human podocalyxin.

    Science.gov (United States)

    Itai, Shunsuke; Yamada, Shinji; Kaneko, Mika K; Kato, Yukinari

    2018-07-01

    Podocalyxin (PODXL) is a type I transmembrane protein, which is highly glycosylated. PODXL is expressed in some types of human cancer tissues including oral, breast, and lung cancer tissues and may promote tumor growth, invasion, and metastasis. We previously produced PcMab-47, a novel anti-PODXL monoclonal antibody (mAb) which reacts with endogenous PODXL-expressing cancer cell lines and normal cells independently of glycosylation in Western blot, flow cytometry, and immunohistochemical analysis. In this study, we used enzyme-linked immunosorbent assay (ELISA), flow cytometry, and immunohistochemical analysis to determine the epitope of PcMab-47. The minimum epitope of PcMab-47 was found to be Asp207, His208, Leu209, and Met210. A blocking peptide containing this minimum epitope completely neutralized PcMab-47 reaction against oral cancer cells by flow cytometry and immunohistochemical analysis. These findings could lead to the production of more functional anti-PODXL mAbs, which are advantageous for antitumor activities.

  7. Simultaneous determination of metformin and vildagliptin in human plasma by a HILIC-MS/MS method.

    Science.gov (United States)

    Pontarolo, Roberto; Gimenez, Ana Carolina; de Francisco, Thais Martins Guimarães; Ribeiro, Rômulo Pereira; Pontes, Flávia Lada Degaut; Gasparetto, João Cleverson

    2014-08-15

    The objective of this work was to develop and validate a HILIC-MS/MS method for the simultaneous determination of metformin and vildagliptin in human plasma. Chromatographic separation was achieved using an Atlantis HILIC Silica 150-mm × 2.1-mm, 3-μm particle size column maintained at 40°C. The isocratic mobile phase consisted of 20% water and 80% acetonitrile/water solution 95:5 (v/v), containing both 0.1% formic acid and 3mM ammonium formate. The flow rate was maintained at 400 μL min(-1). Data from validation studies demonstrated that the new method is highly selective, sensitive (limits of detection vildagliptin in plasma volunteers who orally received a single dose of metformin (850 mg), vildagliptin (50mg) or drug association (metformin 850 mg+vildagliptin 50mg). The new method can thus also be used as a tool for the clinical monitoring of metformin and vildagliptin. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. Synthesis of peptide templated copper nanoclusters for fluorometric determination of Fe(III) in human serum

    International Nuclear Information System (INIS)

    Tang, Ting; Ouyang, Jiang; Hu, Lanshuang; Guo, Linyan; Yang, Minghui; Chen, Xiang

    2016-01-01

    Copper nanoclusters (Cu-NCs) were prepared by reducing CuCl 2 with ascorbic acid in the presence of the short peptide template Cys-Cys-Cys-Asp-Leu. They were characterized by UV-vis absorption and fluorescence spectroscopy, transmission electron microscopy and X-ray photoelectron spectroscopy. The Cu-NCs have a size of ∼2 nm, can be well dispersed in water and are photostable. Their fluorescence (peaking at 425 nm under 365-nm excitation) is quenched by Fe(III) ions. Based on this finding, a sensitive and selective fluorescence assay for the detection of Fe(III) was developed. Under optimized conditions and a pH value of 2.0, the assay displays a linear response in the 0.05 to 30 μM Fe(III) concentration range, with a detection limit of 20 nM based on an S/N ratio of 3. The assay was successfully applied to the determination of Fe(III) in spiked human serum where is gave recoveries that ranged from 96.2 % to 98.3 %. (author)

  9. The determination of chromium-50 in human blood and its utilization for blood volume measurements

    International Nuclear Information System (INIS)

    Zeisler, R.; Young, I.

    1986-01-01

    Possible relationships between insufficient blood volume increases during pregnancy and infant mortality could be established with an adequate measurement procedure. An accurate and precise technique for blood volume measurements has been found in the isotope dilution technique using chromium-51 as a label for red blood cells. However, in a study involving pregnant women, only stable isotopes can be used for labeling. Stable chromium-50 can be determined in total blood samples before and after dilution experiments by neutron activation analysis (NAA) or mass spectrometry. However, both techniques may be affected by insufficient sensitivity and contamination problems at the inherently low natural chromium concentrations to be measured in the blood. NAA procedures involving irradiations with highly thermalized neutrons at a fluence rate of 2x10 13 n/cm 2 xs and low background gamma spectrometry are applied to the analysis of total blood. Natural levels of chromium-50 in human and animal blood have been found to be <0.1 ng/mL; i.e., total chromium levels of <3 ng/mL. Based on the NAA procedure, a new approach to the blood volume measurement via chromium-50 isotope dilution has been developed which utilizes the ratio of the induced activities of chromium-51 to the iron-59 in three blood samples taken from each individual, namely blank, labeled and diluted labeled blood. (author)

  10. Determination of oxidation state of iron in normal and pathologically altered human aortic valves

    Energy Technology Data Exchange (ETDEWEB)

    Czapla-Masztafiak, J. [Institute of Nuclear Physics PAN, Radzikowskiego 152, 31-342 Kraków (Poland); Lis, G.J.; Gajda, M.; Jasek, E. [Department of Histology, Jagiellonian University Medical College, Kopernika 7, 31-034 Kraków (Poland); Czubek, U. [Department of Coronary Disease, Jagiellonian University Medical College, John Paul II Hospital, Prądnicka 80, 31-202 Kraków (Poland); Bolechała, F. [Department of Forensic Medicine, Jagiellonian University Medical College, Grzegórzecka 16, 31-531 Kraków (Poland); Borca, C. [Swiss Light Source, Paul Scherrer Institute, 5232 Villigen PSI (Switzerland); Kwiatek, W.M. [Institute of Nuclear Physics PAN, Radzikowskiego 152, 31-342 Kraków (Poland)

    2015-12-01

    In order to investigate changes in chemical state of iron in normal and pathologically altered human aortic valves X-ray absorption spectroscopy was applied. Since Fe is suspected to play detrimental role in aortic valve stenosis pathogenesis the oxidation state of this element has been determined. The experimental material consisted of 10 μm sections of valves excised during routine surgery and from autopsies. The experiment was performed at the MicroXAS beamline of the SLS synchrotron facility in Villigen (Switzerland). The Fe K-edge XANES spectra obtained from tissue samples were carefully analyzed and compared with the spectra of reference compounds containing iron in various chemical structures. The analysis of absorption edge position and shape of the spectra revealed that both chemical forms of iron are presented in valve tissue but Fe{sup 3+} is the predominant form. Small shift of the absorption edge toward higher energy in the spectra from stenotic valve samples indicates higher content of the Fe{sup 3+} form in pathological tissue. Such a phenomenon suggests the role of Fenton reaction and reactive oxygen species in the etiology of aortic valve stenosis. The comparison of pre-edge regions of XANES spectra for control and stenotic valve tissue confirmed no differences in local symmetry or spin state of iron in analyzed samples.

  11. A modified assay method for determining serotonin uptake in human platelets

    International Nuclear Information System (INIS)

    Arora, R.C.; Meltzer, H.Y.

    1981-01-01

    Effects of various experimental conditions on serotonin (5-HT) uptake in human platelets were examined. The experimental design allowed the evaluation of the effect of diffusion and other non-saturable processes on the affinity and maximum activity of the membrane pump for 5-HT uptake. Total 5-HT uptake was determined by incubating platelet-rich plasma (PRP) with increasing concentrations of serotonin at 37 0 C for 4 min. The passive uptake was measured by the addition of various 5-HT concentrations to PRP in buffer at 37 0 C, followed by immediate transfer to an ice-cold water bath. The difference between the total and passive uptake was linear for 6 min. The affinity (Ksub(m)) for active platelet serotonin uptake was 0.45 +- 0.09 μmol/l and maximal rate of uptake (V) was 10.7 +- 2.1 pmol/10 7 platelets/min. The described method provides a convenient and reliable measure of active 5-HT uptake suitable for clinical investigation. The effect of passive diffusion on kinetic parameters is discussed. (Auth.)

  12. Fractionation of human serum lipoproteins and simultaneous enzymatic determination of cholesterol and triglycerides

    Energy Technology Data Exchange (ETDEWEB)

    Qureshi, Rashid Nazir [Polymer-Analysis Group, van' t Hoff Institute for Molecular Sciences, University of Amsterdam, Nieuwe Achtergracht 166, 1018WV Amsterdam (Netherlands); Kok, Wim Th., E-mail: W.Th.Kok@uva.nl [Polymer-Analysis Group, van' t Hoff Institute for Molecular Sciences, University of Amsterdam, Nieuwe Achtergracht 166, 1018WV Amsterdam (Netherlands); Schoenmakers, Peter J. [Polymer-Analysis Group, van' t Hoff Institute for Molecular Sciences, University of Amsterdam, Nieuwe Achtergracht 166, 1018WV Amsterdam (Netherlands)

    2009-11-03

    A method based on Asymmetric Flow Field-Flow Fractionation (AF4) was developed to separate different types of lipoproteins from human serum. The emphasis in the method optimization was on the possibilities to characterize the largest lipoprotein fractions (LDL and VLDL), which is usually not possible with the size-exclusion chromatography methods applied in routine analysis. Different channel geometries and flow programs were tested and compared. The use of a short fractionation channel was shown to give less sample dilution at the same fractionation power compared to a conventional, long channel. Different size selectivities were obtained with an exponential decay and a linear cross flow program. The ratio of the UV absorption signal to the light scattering signal was used to validate the relation between retention time and size of the fractionated particles. An experimental setup was developed for the simultaneous determination of the cholesterol and triglycerides distribution over the lipoprotein fractions, based on enzymatic reactions followed by UV detection at 500 nm. Coiled and knitted PTFE tubing reactors were compared. An improved peak sharpness and sensitivity were observed with the knitted tubing reactor. After optimization of the experimental conditions a satisfactory linearity and precision (2-3% rsd for cholesterol and 5-6% rsd for triglycerides) were obtained. Finally, serum samples, a pooled sample from healthy volunteers and samples of sepsis patients, were analyzed with the method developed. Lipoprotein fractionation and cholesterol and triglyceride distributions could be correlated with the clinical background of the samples.

  13. OPTIMIZATION AND VALIDATION OF HPLC METHOD FOR TETRAMETHRIN DETERMINATION IN HUMAN SHAMPOO FORMULATION.

    Science.gov (United States)

    Zeric Stosic, Marina Z; Jaksic, Sandra M; Stojanov, Igor M; Apic, Jelena B; Ratajac, Radomir D

    2016-11-01

    High-performance liquid chromatography (HPLC) method with diode array detection (DAD) were optimized and validated for separation and determination of tetramethrin in an antiparasitic human shampoo. In order to optimize separation conditions, two different columns, different column oven temperatures, as well as mobile phase composition and ratio, were tested. Best separation was achieved on the Supelcosil TM LC-18- DB column (4.6 x 250 mm), particle size 5 jim, with mobile phase methanol : water (78 : 22, v/v) at a flow rate of 0.8 mL/min and at temperature of 30⁰C. The detection wavelength of the detector was set at 220 nm. Under the optimum chromatographic conditions, standard calibration curve was measured with good linearity [r2 = 0.9997]. Accuracy of the method defined as a mean recovery of tetramethrin from shampoo matrix was 100.09%. The advantages of this method are that it can easily be used for the routine analysis of drug tetramethrin in pharmaceutical formulas and in all pharmaceutical researches involving tetramethrin.

  14. [Determination of the healing effect of Piper aduncum (spiked pepper or matico) on human fibroblasts].

    Science.gov (United States)

    Paco, Karen; Ponce-Soto, Luis Alberto; Lopez-Ilasaca, Marco; Aguilar, José L

    2016-01-01

    To evaluate the healing effect of a Piper aduncum ethanol-water extract on an adult human dermal fibroblast cell line (hDFa). After obtaining the extract via solid-liquid extraction, concentration, and lyophilization, extract proteins were purified using reverse phase high-performance liquid chromatography, identified using tandem mass spectrometry of tryptic peptides, and analyzed using MALDI-TOF-TOF on an ABSciex4800 mass spectrometer. Half maximum effective concentration values (EC50), half maximum inhibiting concentration (IC50), and percentages of cell proliferation were determined using tetrazolium salt assays. Cell migration was evaluated using a "scratch assay". Growth factor expression in cells was analyzed via quantitative real-time reverse transcription polymerase chain reaction. Against the hDFa cell line, the extract had an IC50 of 200 μg/mL and EC50 of 103.5 µg/mL. In the proliferation assay, protein K2 (obtained from the extract) exhibited increased proliferative activity relative to other treatments (1 µg/mL); this agent also exhibited increased activity (50 µg/mL) in the fibroblast migration assay.Furthermore, the relative expression of platelet-derived growth factor increased by 8.6-fold in the presence of K2 protein relative to the control. The hydroethanolic extract of Piper aduncum and its component proteins increased the proliferation and migration of hDFa and increased the expression of growth factors involved in the healing process.

  15. Identification of virulence determinants of the human pathogenic fungi Aspergillus fumigatus and Candida albicans by proteomics.

    Science.gov (United States)

    Kniemeyer, Olaf; Schmidt, André D; Vödisch, Martin; Wartenberg, Dirk; Brakhage, Axel A

    2011-06-01

    Both fungi Candida albicans and Aspergillus fumigatus can cause a number of life-threatening systemic infections in humans. The commensal yeast C. albicans is one of the main causes of nosocomial fungal infectious diseases, whereas the filamentous fungus A. fumigatus has become one of the most prevalent airborne fungal pathogens. Early diagnosis of these fungal infections is challenging, only a limited number of antifungals for treatment are available, and the molecular details of pathogenicity are hardly understood. The completion of both the A. fumigatus and C. albicans genome sequence provides the opportunity to improve diagnosis, to define new drug targets, to understand the functions of many uncharacterised proteins, and to study protein regulation on a global scale. With the application of proteomic tools, particularly two-dimensional gel electrophoresis and LC/MS-based methods, a comprehensive overview about the proteins of A. fumigatus and C. albicans present or induced during environmental changes and stress conditions has been obtained in the past 5 years. However, for the discovery of further putative virulence determinants, more sensitive and targeted proteomic methods have to be applied. Here, we review the recent proteome data generated for A. fumigatus and C. albicans that are related to factors required for pathogenicity. Copyright © 2011 Elsevier GmbH. All rights reserved.

  16. Elasticity of human embryonic stem cells as determined by atomic force microscopy.

    Science.gov (United States)

    Kiss, Robert; Bock, Henry; Pells, Steve; Canetta, Elisabetta; Adya, Ashok K; Moore, Andrew J; De Sousa, Paul; Willoughby, Nicholas A

    2011-10-01

    The expansive growth and differentiation potential of human embryonic stem cells (hESCs) make them a promising source of cells for regenerative medicine. However, this promise is off set by the propensity for spontaneous or uncontrolled differentiation to result in heterogeneous cell populations. Cell elasticity has recently been shown to characterize particular cell phenotypes, with undifferentiated and differentiated cells sometimes showing significant differences in their elasticities. In this study, we determined the Young's modulus of hESCs by atomic force microscopy using a pyramidal tip. Using this method we are able to take point measurements of elasticity at multiple locations on a single cell, allowing local variations due to cell structure to be identified. We found considerable differences in the elasticity of the analyzed hESCs, reflected by a broad range of Young's modulus (0.05-10 kPa). This surprisingly high variation suggests that elasticity could serve as the basis of a simple and efficient large scale purification/separation technique to discriminate subpopulations of hESCs.

  17. A validated HPLC determination of the flavone aglycone diosmetin in human plasma.

    Science.gov (United States)

    Kanaze, Feras Imad; Bounartzi, Melpomeni I; Niopas, Ioannis

    2004-12-01

    Diosmetin, 3',5,7-trihydroxy-4'-methoxy flavone, is the aglycone of the flavonoid glycoside diosmin that occurs naturally in foods of plant origin. Diosmin exhibits antioxidant and anti-inflammatory activities, improves venous tone and it is used for the treatment of chronic venous insufficiency. Diosmin is hydrolyzed by enzymes of intestinal micro flora before absorption of its aglycone diosmetin. A specific, sensitive, precise, accurate and robust HPLC assay for the determination of diosmetin in human plasma was developed and validated. Diosmetin and the internal standard 7-ethoxycoumarin were isolated from plasma by liquid-liquid extraction and separated on a C8 reversed-phase column with methanol-water-acetic acid (55:43:2, v/v/v) as the mobile phase at 43 degrees C. Peaks were monitored at 344 nm. The method was linear in the 10-300 ng/mL concentration range (r > 0.999). Recovery for diosmetin and internal standard was greater than 89.7 and 86.8%, respectively. Intra-day and inter-day precision for diosmetin ranged from 1.6 to 4.6 and from 2.2 to 5.3%, respectively, and accuracy was better than 97.9%. Copyright 2004 John Wiley & Sons, Ltd.

  18. Analytical method by high resolution liquid chromatography for the determination of carbamazepine in human plasma

    International Nuclear Information System (INIS)

    Jimenez Aleman, Narda M; Calero Carbonell, Jorge E; Padron Yaquis, Alejandro S; Izquierdo Lozano, Julio C

    2007-01-01

    One of the requirements to develop the studies of bioavailability and bioequivalence is to have analytic methodologies validated for the work with samples in biological fluids. A method was developed by high resolution liquid chromatography for the determination of carbamazepine in human plasma. A mixture of hydrogen phosphate of sodium: acetonitrile (65:35) adjusted to pH= 3.3 with phosphoric acid, flow of 1.2 mL/min and ultraviolet detection at 210 nm, was used as mobile phase. Propylparabene was used as an internal standard. According to the established regulations for the validation of the methods in biological fluids, the following parameters were studied: stability of the samples, lineality, specificity, precision, accuracy and limit of detection and quantification. The method proved to be specific and sensitive with a detection and quantification limit of 0.9 and 1.0 ng, respectively. The method was lineal, precise and exact in the range of concentrations of 1. 07 at 12.67 μg/mL. The mean recovery was not statistically different from 100.0 %. The analito in the proposed biological matrix remained in the studied period. The methodology described in this work is applied in our case to the study that evaluates the bioavailability and bioequivalence of a Cuban formulation of carbamazepine in healthy volunteers. (Author)

  19. Extraction and Determination of Cyproheptadine in Human Urine by DLLME-HPLC Method.

    Science.gov (United States)

    Maham, Mehdi; Kiarostami, Vahid; Waqif-Husain, Syed; Abroomand-Azar, Parviz; Tehrani, Mohammad Saber; Khoeini Sharifabadi, Malihe; Afrouzi, Hossein; Shapouri, Mahmoudreza; Karami-Osboo, Rouhollah

    2013-01-01

    Novel dispersive liquid-liquid microextraction (DLLME), coupled with high performance liquid chromatography with photodiode array detection (HPLC-DAD) has been applied for the extraction and determination of cyproheptadine (CPH), an antihistamine, in human urine samples. In this method, 0.6 mL of acetonitrile (disperser solvent) containing 30 μL of carbon tetrachloride (extraction solvent) was rapidly injected by a syringe into 5 mL urine sample. After centrifugation, the sedimented phase containing enriched analyte was dissolved in acetonitrile and an aliquot of this solution injected into the HPLC system for analysis. Development of DLLME procedure includes optimization of some important parameters such as kind and volume of extraction and disperser solvent, pH and salt addition. The proposed method has good linearity in the range of 0.02-4.5 μg mL(-1) and low detection limit (13.1 ng mL(-1)). The repeatability of the method, expressed as relative standard deviation was 4.9% (n = 3). This method has also been applied to the analysis of real urine samples with satisfactory relative recoveries in the range of 91.6-101.0%.

  20. Structure determination of human Lck unique and SH3 domains by nuclear magnetic resonance spectroscopy

    Directory of Open Access Journals (Sweden)

    Willbold Dieter

    2003-05-01

    Full Text Available Abstract Background Protein tyrosine kinases are involved in signal transduction pathways that regulate cell growth, differentiation, activation and transformation. Human lymphocyte specific kinase (Lck is a 56 kDa protein involved in T-cell- and IL2-receptor signaling. Three-dimensional structures are known for SH3, SH2 and kinase domains of Lck as well as for other tyrosine kinases. No structure is known for the unique domain of any Src-type tyrosine kinase. Results Lck(1–120 comprising unique and SH3 domains was structurally investigated by nuclear magnetic resonance spectroscopy. We found the unique domain, in contrast to the SH3 part, to have basically no defined structural elements. The solution structure of the SH3 part could be determined with very high precision. It does not show significant differences to Lck SH3 in the absence of the unique domain. Minor differences were observed to the X-ray structure of Lck SH3. Conclusion The unique domain of Lck does not contain any defined structure elements in the absence of ligands and membranes. Presence of the unique domain is not relevant to the three-dimensional structure of the Lck SH3 domain.

  1. Determination of nitrite/nitrate in human biological material by the simple Griess reaction.

    Science.gov (United States)

    Guevara, I; Iwanejko, J; Dembińska-Kieć, A; Pankiewicz, J; Wanat, A; Anna, P; Gołabek, I; Bartuś, S; Malczewska-Malec, M; Szczudlik, A

    1998-06-22

    Since a number of pathological processes such as septic shock, inflammation, graft rejection, diabetes, etc. are associated with a release of nitric oxide (NO), rapid and accurate methods of monitoring of NO concentration are of interest. Various methods for measurement of nitrite and nitrate (NO2-, NO3- ) -- the stable metabolites of NO -- are commonly used for this purpose. In this paper we have shown that the proper Griess procedure for nitrite determination significantly increases the sensitivity of this method. This procedure, supplemented with deproteinization and reduction of nitrates to nitrites in the presence of NADPH-sensitive reductase, can be successfully applied for measurement of NOx levels in human body fluids (serum, urine and CSF). Deproteinization of samples with methanol/diethylether is required and does not influence the sensitivity of detection of NO metabolites. The recovery of the method is 88%+/-6% (n = 30). The NOx concentrations measured by this procedure ranged from 25.0 to 39.0 micromol/l in blood, 4.6 to 14.6 micromol/l in CSF and 0.37 to 2.52 mmol/l (adjusted to creatinine concentration) in urine. The coefficient of variation for this method was between 1.3-2.2%. This method can also be recommended for measurement of NOx produced by cells in tissue cell culture.

  2. In vivo determination of the absorption and scattering spectra of the human prostate during photodynamic therapy

    Science.gov (United States)

    Finlay, Jarod C.; Zhu, Timothy C.; Dimofte, Andreea; Stripp, Diana C. H.; Malkowicz, S. B.; Whittington, Richard; Miles, Jeremy; Glatstein, Eli; Hahn, Stephen M.

    2004-06-01

    A continuing challenge in photodynamic therapy is the accurate in vivo determination of the optical properties of the tissue being treated. We have developed a method for characterizing the absorption and scattering spectra of prostate tissue undergoing PDT treatment. Our current prostate treatment protocol involves interstitial illumination of the organ via cylindrical diffusing optical fibers (CDFs) inserted into the prostate through clear catheters. We employ one of these catheters to insert an isotropic white light point source into the prostate. An isotropic detection fiber connected to a spectrograph is inserted into a second catheter a known distance away. The detector is moved along the catheter by a computer-controlled step motor, acquiring diffuse light spectra at 2 mm intervals along its path. We model the fluence rate as a function of wavelength and distance along the detector"s path using an infinite medium diffusion theory model whose free parameters are the absorption coefficient μa at each wavelength and two variables A and b which characterize the reduced scattering spectrum of the form μ"s = Aλ-b. We analyze our spectroscopic data using a nonlinear fitting algorithm to determine A, b, and μa at each wavelength independently; no prior knowledge of the absorption spectrum or of the sample"s constituent absorbers is required. We have tested this method in tissue simulating phantoms composed of intralipid and the photosensitizer motexafin lutetium (MLu). The MLu absorption spectrum recovered from the phantoms agrees with that measured in clear solution, and μa at the MLu absorption peak varies linearly with concentration. The ´"s spectrum reported by the fit is in agreement with the known scattering coefficient of intralipid. We have applied this algorithm to spectroscopic data from human patients sensitized with MLu (2 mg kg-1) acquired before and after PDT. Before PDT, the absorption spectra we measure include the characteristic MLu absorption

  3. Sensitive cell-based assay for determination of human immunodeficiency virus type 1 coreceptor tropism.

    Science.gov (United States)

    Weber, Jan; Vazquez, Ana C; Winner, Dane; Gibson, Richard M; Rhea, Ariel M; Rose, Justine D; Wylie, Doug; Henry, Kenneth; Wright, Alison; King, Kevin; Archer, John; Poveda, Eva; Soriano, Vicente; Robertson, David L; Olivo, Paul D; Arts, Eric J; Quiñones-Mateu, Miguel E

    2013-05-01

    CCR5 antagonists are a powerful new class of antiretroviral drugs that require a companion assay to evaluate the presence of CXCR4-tropic (non-R5) viruses prior to use in human immunodeficiency virus (HIV)-infected individuals. In this study, we have developed, characterized, verified, and prevalidated a novel phenotypic test to determine HIV-1 coreceptor tropism (VERITROP) based on a sensitive cell-to-cell fusion assay. A proprietary vector was constructed containing a near-full-length HIV-1 genome with the yeast uracil biosynthesis (URA3) gene replacing the HIV-1 env coding sequence. Patient-derived HIV-1 PCR products were introduced by homologous recombination using an innovative yeast-based cloning strategy. The env-expressing vectors were then used in a cell-to-cell fusion assay to determine the presence of R5 and/or non-R5 HIV-1 variants within the viral population. Results were compared with (i) the original version of Trofile (Monogram Biosciences, San Francisco, CA), (ii) population sequencing, and (iii) 454 pyrosequencing, with the genotypic data analyzed using several bioinformatics tools, i.e., the 11/24/25 rule, Geno2Pheno (2% to 5.75%, 3.5%, or 10% false-positive rate [FPR]), and webPSSM. VERITROP consistently detected minority non-R5 variants from clinical specimens, with an analytical sensitivity of 0.3%, with viral loads of ≥1,000 copies/ml, and from B and non-B subtypes. In a pilot study, a 73.7% (56/76) concordance was observed with the original Trofile assay, with 19 of the 20 discordant results corresponding to non-R5 variants detected using VERITROP and not by the original Trofile assay. The degree of concordance of VERITROP and Trofile with population and deep sequencing results depended on the algorithm used to determine HIV-1 coreceptor tropism. Overall, VERITROP showed better concordance with deep sequencing/Geno2Pheno at a 0.3% detection threshold (67%), whereas Trofile matched better with population sequencing (79%). However, 454

  4. CCQM-K11.2 determination of glucose in human serum and CCQM-K12.2 determination of creatinine in human serum

    Science.gov (United States)

    Wise, Stephen A.; Phinney, Karen W.; Duewer, David L.; Sniegoski, Lorna T.; Welch, Michael J.; Pritchett, Jeanita; Pabello, Guiomar; Avila Calderon, Marco A.; Balderas, Miryan; Qinde, Liu; Kooi, Lee Tong; Rego, Eliane; Garrido, Bruno; Allegri, Gabriella; de La Cruz, Marcia; Barrabin, Juliana; Monteiro, Tânia; Lee, Hwashim; Kim, Byungjoo; Delatour, Vincent; Peignaux, Maryline; Kawaguchi, Migaku; Bei, Xu; Can, Quan; Nammoonnoy, Jintana; Schild, Katrin; Ohlendorf, Rüdiger; Henrion, Andre; Ceyhan Gören, Ahmet; Yılmaz, Hasibe; Bilsel, Mine; Konopelko, L.; Krylov, A.; Lopushanskaya, E.

    2018-01-01

    Glucose and creatinine are two of the most frequently measured substances in human blood/serum for assessing the health status of individuals. Because of their clinical significance, CCQM-K11 glucose in human serum and CCQM-K12 creatinine in human serum were the fourth and fifth key comparisons (KCs) performed by the Organic Analysis Working Group (OAWG). These KCs were conducted in parallel and were completed in 2001. The initial subsequent KCs for glucose, CCQM-K11.1, and creatinine, CCQM-K12.1, were completed in 2005. Measurements for the next KCs for these two measurands, CCQM-K11.2 and CCQM-K12.2, were completed in 2013. While designed as subsequent KCs, systematic discordances between the participants' and the anchor institution's results in both comparisons lead the OAWG to request reference results from two experienced laboratories that had participated in the 2001 comparisons. Based on the totality of the available information, the OAWG converted both CCQM-K11.2 and CCQM-K12.2 to 'Track C' KCs where the key comparison reference value is estimated by consensus. These comparisons highlighted that carrying out comparisons for complex chemical measurements and expecting to be able to treat them under the approaches used for formal CIPM subsequent comparisons is not an appropriate strategy. The approach used here is a compromise to gain the best value from the comparison; it is not an approach that will be used in the future. Instead, the OAWG will focus on Track A and Track C comparisons that are treated as stand-alone entities. Participation in CCQM-K11.2 demonstrates a laboratory's capabilities to measure a polar (pKow > 2), low molecular mass (100 g/mol to 500 g/mol) metabolite in human serum at relatively high concentrations (0.1 mg/g to 10 mg/g). Participation in CCQM-K12.2 demonstrates capabilities to measure similar classes of metabolites at relatively low concentrations (1 μg/g to 30 μg/g). The capabilities required for the analysis of complex

  5. Drinking behavior in nursery pigs: determining the accuracy between an automatic water meter versus human observers.

    Science.gov (United States)

    Meiszberg, A M; Johnson, A K; Sadler, L J; Carroll, J A; Dailey, J W; Krebs, N

    2009-12-01

    Assimilating accurate behavioral events over a long period can be labor-intensive and relatively expensive. If an automatic device could accurately record the duration and frequency for a given behavioral event, it would be a valuable alternative to the traditional use of human observers for behavioral studies. Therefore, the objective of this study was to determine the accuracy in the time spent at the waterer and the number of visits to the waterer by individually housed nursery pigs between human observers scoring video files using Observer software (OBS) and an automatic water meter Hobo (WM, control) affixed onto the waterline. Eleven PIC USA genotype gilts (22 +/- 2 d of age; 6.5 +/- 1.4 kg of BW) were housed individually in pens with ad libitum access to a corn-based starter ration and one nipple waterer. Behavior was collected on d 0 (day of weaning), 7, and 14 of the trial using 1 color camera positioned over 4 attached pens and a RECO-204 DVR at 1 frame per second. For the OBS method, 2 experienced observers recorded drinking behavior from the video files, which was defined as when the gilt placed her mouth over the nipple waterer. Data were analyzed using nonparametric methods and the general linear model and regression procedures in SAS. The experimental unit was the individual pen housing 1 gilt. The GLM model included the method of observation (WM vs. OBS) and time (24 h) as variables, and the gilt nested within method was used as the error term. Gilts consumed more water (P = 0.04) on d 14 than on d 0. The time of day affected (P waterer regardless of the method. However, the OBS method underestimated (P waterer (3.48 +/- 0.33 visits/h for OBS vs. 4.94 +/- 0.33 for WM) and overestimated (P waterer (22.6 +/- 1.46 s/h for OBS vs. 13.9 +/- 1.43 for WM) compared with WM. The relationship between the 2 methods for prediction of time spent at the waterer and number of visits made by the gilts was weak (R(2) = 0.56 and 0.69, respectively). Collectively

  6. CCQM K6.2 determination of total cholesterol in human serum

    Science.gov (United States)

    Wise, Stephen A.; Phinney, Karen W.; Duewer, David L.; Sniegoski, Lorna T.; Welch, Michael J.; Pabello, Guiomar; Avila Caldero, Marco A.; Qinde, Liu; Kooi, Lee Tong; Rego, Eliane; Garrido, Bruno; Allegri, Gabriella; de La Cruz, Marcia; Barrabin, Juliana; Puglisi, Celia; Lopez, Eduardo; Lee, Hwashim; Kim, Byungjoo; Delatour, Vincent; Heuillet, Maud; Nammoonnoy, Jintana; Ceyhan Gören, Ahmet; Bilsel, Gokhan; Konopelko, L.; Krylov, A.; Lopushanskaya, E.

    2018-01-01

    Cholesterol is one of the most frequently measured substances in human blood/serum to assist in assessing the health status of individuals. Because of its clinical significance, CCQM-K6 determination of cholesterol in serum was completed in 2000 as one of the first key comparison (KC) studies performed within the Organic Analysis Working Group (OAWG). The first subsequent KC for cholesterol, CCQM-K6.1, was completed in 2001. Measurements for this second subsequent, CCQM-K6.2, were completed in 2012. These subsequent comparisons were conducted to enable CCQM members that had not participated in earlier studies to demonstrate their capabilities to measure a nonpolar (pKow mass (100 g/mol to 500 g/mol) metabolite in human serum at relatively high concentrations (1 mg/g to 3 mg/g) found in normal populations. Successful participation in CCQM-K6.2 demonstrated capabilities in analysis of complex biological matrices including sample preparation (extraction, derivatization), LC or GC separation, and quantification using an isotope dilution mass spectrometry approach. Normally in a subsequent KC, no key comparison reference value (KCRV) would be established and assessment of performance would be via the deviation of participants' results to the anchor institute's results, adjusted to account for the anchor's performance in the original comparison versus its KCRV. Due to the very long-time period since the original key comparison, the OAWG decided that this did not represent the best approach to assess performance in what is a relatively complex measurement. Given the excellent agreement between the anchor institute's results and robust consensus summary of the participants' values, the reference value for this study was taken as the anchor institute's result and treated as a 'KCRV'. Seven of the nine participants demonstrated agreement with the reference value. Main text To reach the main text of this paper, click on Final Report. Note that this text is that which appears

  7. Determination of salbutamol and salbutamol glucuronide in human urine by means of liquid chromatography-tandem mass spectrometry

    DEFF Research Database (Denmark)

    Mareck, Ute; Guddat, Sven; Schwenke, Anne

    2012-01-01

    The determination of salbutamol and its glucuronide in human urine following the inhalative and oral administration of therapeutic doses of salbutamol preparations was performed by means of direct urine injection utilizing liquid chromatography-tandem mass spectrometry (LC-MS/MS) and employing d(3...... glucuronide values between 8 and 15 ng/ml. The approach enabled the rapid determination of salbutamol and its glucuronic acid conjugate in human urine and represents an alternative to existing procedures since time-consuming hydrolysis or derivatization steps were omitted. Moreover, the excretion...

  8. Role of Notch signaling in cell-fate determination of human mammary stem/progenitor cells

    International Nuclear Information System (INIS)

    Dontu, Gabriela; Jackson, Kyle W; McNicholas, Erin; Kawamura, Mari J; Abdallah, Wissam M; Wicha, Max S

    2004-01-01

    Notch signaling has been implicated in the regulation of cell-fate decisions such as self-renewal of adult stem cells and differentiation of progenitor cells along a particular lineage. Moreover, depending on the cellular and developmental context, the Notch pathway acts as a regulator of cell survival and cell proliferation. Abnormal expression of Notch receptors has been found in different types of epithelial metaplastic lesions and neoplastic lesions, suggesting that Notch may act as a proto-oncogene. The vertebrate Notch1 and Notch4 homologs are involved in normal development of the mammary gland, and mutated forms of these genes are associated with development of mouse mammary tumors. In order to determine the role of Notch signaling in mammary cell-fate determination, we have utilized a newly described in vitro system in which mammary stem/progenitor cells can be cultured in suspension as nonadherent 'mammospheres'. Notch signaling was activated using exogenous ligands, or was inhibited using previously characterized Notch signaling antagonists. Utilizing this system, we demonstrate that Notch signaling can act on mammary stem cells to promote self-renewal and on early progenitor cells to promote their proliferation, as demonstrated by a 10-fold increase in secondary mammosphere formation upon addition of a Notch-activating DSL peptide. In addition to acting on stem cells, Notch signaling is also able to act on multipotent progenitor cells, facilitating myoepithelial lineage-specific commitment and proliferation. Stimulation of this pathway also promotes branching morphogenesis in three-dimensional Matrigel cultures. These effects are completely inhibited by a Notch4 blocking antibody or a gamma secretase inhibitor that blocks Notch processing. In contrast to the effects of Notch signaling on mammary stem/progenitor cells, modulation of this pathway has no discernable effect on fully committed, differentiated, mammary epithelial cells. These studies

  9. Determination of Scopolamine in Human Saliva Using Solid Phase Extraction and LC/MS/MS

    Science.gov (United States)

    Wang, Zuwei; Vaksman, Zalman; Boyd, Jason; Putcha, Lakshmi

    2007-01-01

    Purpose: Scopolamine is the preferred treatment for motion sickness during space flight because of its quick onset of action, short half-life and favorable side-effect profile. The dose administered depends on the mode of administration and usually ranges between 0.1 and 0.8 mg. Such small doses make it difficult to detect concentrations of scopolamine in biological fluids by using conventional HPLC methods. To measure scopolamine in saliva and thereby to evaluate the pharmacokinetics of scopolamine, we developed an LC/MS/MS method using off-line solid phase extraction. Method: Samples (0.5mL) were loaded onto Waters Oasis HLB co-polymer cartridges (10 mg, 1 mL) and eluted with 0.5 mL methanol without evaporation and reconstitution. HPLC separation of the eluted sample was performed using an Agilent Zorbax SB-CN column (50 x 2.1 mm) at a flow rate of 0.2 mL/min for 4 minutes. The mobile phase for separation was 90:10 (v/v) methanol: ammonium acetate (2 mM) in water, pH 5.0 +/- 0.1. Concentrations of scopolamine were determined using a Micromass Quattro Micro(TM) mass spectrometer with electrospray ionization (ESI). ESI mass spectra were acquired in positive ion mode with multiple reaction monitoring for the determination of scopolamine m/z = 304.2 yields 138.1 and internal standard (IS) hyoscyamine m/z = 290.2 yields 124.1. Results: The method is rapid, reproducible, specific and has the following parameters: scopolamine and the IS are eluted at 1.7 and 3.2 min respectively. The linear range is 50-5000 pg/mL for scopolamine in saliva with correlation coefficients > 0.99 with a CV < 0.5 %. The intra-day and inter-day CVs are < 15 % for quality control samples with concentrations of 75, 300, 750 and 3000 pg/mL of scopolamine in human saliva. Conclusion: Solid phase extraction allows more rapid sample preparation and greater precision than liquid extraction. Furthermore, we increased the sensitivity and specificity by adjusting the LC mobile phase and using an MS

  10. Determining the binding affinity of therapeutic monoclonal antibodies towards their native unpurified antigens in human serum.

    Directory of Open Access Journals (Sweden)

    Christine Bee

    Full Text Available Monoclonal antibodies (mAbs are a growing segment of therapeutics, yet their in vitro characterization remains challenging. While it is essential that a therapeutic mAb recognizes the native, physiologically occurring epitope, the generation and selection of mAbs often rely on the use of purified recombinant versions of the antigen that may display non-native epitopes. Here, we present a method to measure both, the binding affinity of a therapeutic mAb towards its native unpurified antigen in human serum, and the antigen's endogenous concentration, by combining the kinetic exclusion assay and Biacore's calibration free concentration analysis. To illustrate the broad utility of our method, we studied a panel of mAbs raised against three disparate soluble antigens that are abundant in the serum of healthy donors: proprotein convertase subtilisin/kexin type 9 (PCSK9, progranulin (PGRN, and fatty acid binding protein (FABP4. We also determined the affinity of each mAb towards its purified recombinant antigen and assessed whether the interactions were pH-dependent. Of the six mAbs studied, three did not appear to discriminate between the serum and recombinant forms of the antigen; one mAb bound serum antigen with a higher affinity than recombinant antigen; and two mAbs displayed a different affinity for serum antigen that could be explained by a pH-dependent interaction. Our results highlight the importance of taking pH into account when measuring the affinities of mAbs towards their serum antigens, since the pH of serum samples becomes increasingly alkaline upon aerobic handling.

  11. Determinants in the uptake of the Human Papillomavirus vaccine: a systematic review based on European studies

    Directory of Open Access Journals (Sweden)

    Victoria eFernández de Casadevante

    2015-06-01

    Full Text Available Background Cervical cancer is the fourth most common cancer affecting women worldwide. Since 2006, two Human Papillomavirus vaccines (HPVV have been licensed to protect women against the virus that causes cervical cancer. However, worldwide coverage remains unequal. Studies from the USA found strong evidence for differences in HPVV uptake by ethnicity and healthcare coverage. As the profile of ethnic groups and the healthcare system in the USA differ from countries in Europe where HPVV is free in most of the countries, we conducted a systematic review in order to analyze the determinants of HPVV uptake in Europe.Methods We performed a systematic Pubmed, Scopus and Science Direct search to find articles published from HPVV availability in European countries until April 2014. No age restriction was applied. We included all studies assessing factors associated with HPVV uptake. Uptake refers to either initiation and/or completion of the three dose vaccination program. Results Out of the 23 eligible studies, 14 were retrospective reviews of data, six were cross-sectional surveys and three were prospective cohort studies. Higher HPVV uptake was associated with ethnic majority populations, higher socio-economic status, regular cervical screening participation by the mother and having received previous childhood vaccinations.Conclusions Since the vaccine is offered for free in most of the European countries, the findings suggest that ethno-cultural and educational factors play an important role when it comes to HPVV uptake. Girls who were undervaccinated had also a lower uptake of standard childhood vaccines and mothers who were less likely to attend cervical cancer screening. This may indicate, that only few parents have specific concerns with HPVV, and that preventive health care should seek ways to target these vulnerable groups.

  12. Development and Validation of a HPLC Method for Determination of Pefloxacin in Tablet and Human Plasma

    Directory of Open Access Journals (Sweden)

    Shahnaz Gauhar

    2009-03-01

    Full Text Available Objective(sDeveloping and validating a simple, efficient, reproducible and economic reversed phase high performance liquid chromatographic (RP-HPLC method for the quantitative determination of pefloxacin in bulk material, tablets and in human plasma. Materials and MethodsA shim-pack CLC-ODS column and a mobile phase constituting acetonitrile: 0.025 M phosphoric acid solution (13:87 v/v, pH 2.9 adjusted with KOH were used. The flow rate was 1 ml/min and the analyses performed using ultraviolet (UV detector at a wavelength of 275 nm using acetaminophen as an internal standard.ResultsThe developed method showed good resolution between pefloxacin and acetaminophen. It was selective to pefloxacin and able to resolve the drug peak from internal standard and from formulation excipients. The percentage of coefficient variation (CV of the retention times and peak areas of pefloxacin from the six consecutive injections were 0.566% and 0.989%, respectively. The results showed that the peak area responses are linear within the concentration range of 0.125 µg/ml-12 µg/ml (R2= 0.9987. The limits of detection (LOD and limits of quantitation (LOQ for pefloxacin were 0.03125 µg/ml and 0.125 µg/ml. The intra-day and inter-day variation, RSD were 0.376-0.9056 and 0.739-0.853 respectively; also, inter-day variation with relative standard deviation (RSD were 0.1465-0.821 in plasma. The accuracy results of 70%, 100%, and 130% drugs were 100.72%, 100.34%, and 100.09%, respectively.ConclusionThe method is linear, quantitative, reproducible and could be used as a more convenient, efficient and economical method for the trace analysis of drug in biological fluids, in raw material and tablets.

  13. Liquid Chromatography-Tandem Mass Spectrometric Assay for Determination of Stavudine in Human Plasma

    Directory of Open Access Journals (Sweden)

    Fengdan Jin

    2014-01-01

    Full Text Available A LC-MS/MS method for determination of stavudine in human plasma was established and validated, and it was applied to the pharmaceutical formulations bioequivalence study. 0.5 mL plasma sample was extracted by liquid-liquid extraction. Stavudine was detected by a LC-MS/MS system. The pharmacokinetic parameters of stavudine in different formulations were calculated by noncompartment model statistics. The method was linear over the concentration ranges 5.00–1000 ng/mL in plasma. The intra- and interassay relative standard deviation (RSD was <10%. The average accuracies for the assay at three concentrations (5.00, 80.0, and 900 ng/mL were from 100.2% to 102.5%. Pharmacokinetic parameters of stavudine reference formulation were obtained as follows: Tmax was 0.6±0.2 h, Cmax was 480.7±150.9 g/L, t1/2 was 1.7±0.4 h, and AUC0-t was 872.8±227.8 g·h/L, and pharmacokinetic parameters of stavudine test formulation were obtained as follows: Tmax was 0.5±0.2 h, Cmax was 537.5±178.5 g/L, t1/2 was 1.7±0.3 h, and AUC0-t was (914.1±284.5 g·h/L. Calculated with AUC0-t, the bioavailability of two formulations was 105.0%.

  14. COMMUNITY DETERMINANTS OF IMMIGRANT SELF-EMPLOYMENT: HUMAN CAPITAL SPILLOVERS AND ETHNIC ENCLAVES

    OpenAIRE

    Liliana Sousa

    2013-01-01

    I find evidence that human capital spillovers have positive effects on the proclivity of low human capital immigrants to self-employ. Human capital spillovers within an ethnic community can increase the self-employment propensity of its members by decreasing the costs associated with starting and running a business (especially, transaction costs and information costs). Immigrants who do not speak English and those with little formal education are more likely to be self-employed if they reside...

  15. Use of magnesium silicate as a selective absorbent in radioimmunological method of determination of insulin level in human serum

    Energy Technology Data Exchange (ETDEWEB)

    Bogoniowska, Z; Stelmasiak, T [Wojskowy Instytut Higieny i Epidemiologii, Warsaw (Poland)

    1974-01-01

    The authors present a radioimmunological method for determination of insulin (IRI) level in the human serum using magnesium silicate (talc) as adsorbent. The method is based on the phenomenon of selective adsorption of the free radioactive hormone. The optimal parameters for the method were determined. The serum level of IRI in clinically healthy subjects after oral glucose loading was established. The obtained results were compared with the results obtained by the radioimmunological method of double antibodies in stochastically grouped samples.

  16. Quantitative determination of the human breast milk macronutrients by near-infrared Raman spectroscopy

    Science.gov (United States)

    Motta, Edlene d. C. M.; Zângaro, Renato A.; Silveira, Landulfo, Jr.

    2012-03-01

    This work proposes the evaluation of the macronutrient constitution of human breast milk based on the spectral information provided by near-infrared Raman spectroscopy. Human breast milk (5 mL) from a subject was collected during the first two weeks of breastfeeding and stocked in -20°C freezer. Raman spectra were measured using a Raman spectrometer (830 nm excitation) coupled to a fiber based Raman probe. Spectra of human milk were dominated by bands of proteins, lipids and carbohydrates in the 600-1800 cm-1 spectral region. Raman spectroscopy revealed differences in the biochemical constitution of human milk depending on the time of breastfeeding startup. This technique could be employed to develop a classification routine for the milk in Human Milk Banking (HMB) depending on the nutritional facts.

  17. Direct solid-phase microextraction combined with gas and liquid chromatography for the determination of lidocaine in human urine

    NARCIS (Netherlands)

    Koster, E.H M; Hofman, N.S K; de Jong, G.J.

    Solid-phase microextraction (SPME) has been combined with gas chromatography (GC) and liquid chromatography (LC) for the determination of lidocaine in human urine. A polydimethylsiloxane (PDMS) coated fibre was directly immersed into buffered urine. Extraction conditions such as time, pH, ionic

  18. XRF and TXRF techniques for multi-element determination of trace elements in whole blood and human hair samples

    International Nuclear Information System (INIS)

    Khuder, A.; Karjou, J.; Sawan, M.Kh.; Bakir, M.A.

    2007-01-01

    XRF and TXRF were established as useful techniques for multi-element analysis of whole blood and human head hair samples. Direct-XRF with different collimation units and different X-ray excitation modes was successfully used for the determination of S, P, K, Ca, Fe, and Br elements in blood samples and K, Ca, Mn, Fe elements in human hair samples. Direct analysis by TXRF was used for the determination of Rb and Sr in digested blood and human hair samples, respectively, while, the co-precipitation method using APDC for TXRF analysis was used for the determination of Ni, Cu, Zn, and Pb elements in both matrices. As a result, the improved XRF and TXRF methods were applied for multi-element determination of elements in whole blood and human hair samples in non-occupational exposed population living in Damascus city. The mean concentrations of analyzed elements in both matrices were on the reported range values for non-occupational population in other countries. (author)

  19. XRF and TXRF techniques for multi-element determination of trace elements in whole blood and human hair samples

    International Nuclear Information System (INIS)

    Khuder, A.; Karjou, J.; Sawan, M.Kh.; Bakir, M.A.

    2008-01-01

    XRF and TXRF were established as useful techniques for multi-element analysis of whole blood and human head hair samples. Direct-XRF with different collimation units and different X-ray excitation modes was successfully used for the determination of S, P, K, Ca, Fe, and Br elements in blood samples and K, Ca, Mn, Fe elements in human hair samples. Direct analysis by TXRF was used for the determination of Rb and Sr in digested blood and human hair samples, respectively, while, the co-precipitation method using APDC for TXRF analysis was used for the determination of Ni, Cu, Zn, and Pb elements in both matrices. As a result, the improved XRF and TXRF methods were applied for multi-element determination of elements in whole blood and human hair samples in non-occupational exposed population living in Damascus city. The mean concentrations of analyzed elements in both matrices were on the reported range values for non-occupational population in other countries. (author)

  20. A Comprehensive Experiment for Molecular Biology: Determination of Single Nucleotide Polymorphism in Human REV3 Gene Using PCR-RFLP

    Science.gov (United States)

    Zhang, Xu; Shao, Meng; Gao, Lu; Zhao, Yuanyuan; Sun, Zixuan; Zhou, Liping; Yan, Yongmin; Shao, Qixiang; Xu, Wenrong; Qian, Hui

    2017-01-01

    Laboratory exercise is helpful for medical students to understand the basic principles of molecular biology and to learn about the practical applications of molecular biology. We have designed a lab course on molecular biology about the determination of single nucleotide polymorphism (SNP) in human REV3 gene, the product of which is a subunit of…

  1. Pomp and Circumstance: University Presidents and the Role of Human Capital in Determining Who Leads U.S. Research Institutions

    Science.gov (United States)

    Singell, Larry D., Jr.; Tang, Hui-Hsuan

    2013-01-01

    While there is wide agreement that leaders matter, little is known regarding the role that human capital plays in determining who becomes one. We exploit unique attributes of the higher education industry to examine if training and academic ability affect the placement of university presidents within the research hierarchy of U.S. institutions.…

  2. A study on chemical element determinations in human nails by neutron activation analysis

    International Nuclear Information System (INIS)

    Sanches, Thalita Pinheiro; Saiki, Mitiko

    2013-01-01

    Nail analyses have been the object of study in order to assess the levels of elements accumulated in the human organism and to use this tissue to monitor environmental and occupational exposure, to evaluate the nutritional status, to verify intoxication by toxic metals and to diagnose or to prevent diseases. Nail analyses present advantages due to easy sample collection, storage, transportation and this tissue provides element level accumulation over time. However, there is controversy regarding the application of nail analysis data due to difficulties to establish reliable reference values or element concentration ranges as control values. The objective of this study was to evaluate the factors that can affect nail element concentrations for further sample analyses of a group of individuals by applying neutron activation analysis (NAA). Fingernails and toenails collected from adult individuals of both genders, aged 18 to 71 years, living in the Sao Paulo Metropolitan Region were cut in small fragments, cleaned and dried for analyses. Samples and element standards were irradiated for 16 h under a thermal neutron flux of about 4.5 x 10 12 n cm -2 s -1 at the IEA-R1 nuclear research reactor followed by gamma ray spectrometry. Element concentrations for As, Br, Ca, Co, Cr, Cs, Fe, K, La, Na, Rb, Sb, Sc, Se and Zn were determined. For quality control of the analytical results, certified reference materials were analysed and the results showed good accuracy and precision with relative errors and relative standard deviations lower than 5.1 % and 11.6 %, respectively. Preliminary assays indicated that the contribution due to impurities from plastic involucres used in the irradiation as well as those from nail polishes is very low and could be considered negligible. Results from the nail sample cleaning process using distinct procedures indicated that HNO 3 solution may cause sample dissolution. Sample homogeneity was verified by analysis of a sample in replicate. A

  3. A study on chemical element determinations in human nails by neutron activation analysis

    International Nuclear Information System (INIS)

    Sanches, Thalita Pinheiro

    2012-01-01

    Nail analyses have been the object of study in order to assess the levels of elements accumulated in the human organism and to use this tissue to monitor environmental and occupational exposure, to evaluate the nutritional status, to verify intoxication by toxic metals and to diagnose or to prevent diseases. Nail analyses present advantages due to easy sample collection, storage, transportation and this tissue provides element level accumulation over time. However, there is controversy regarding the application of nail analysis data due to difficulties to establish reliable reference values or element concentration ranges as control values. The objective of this study was to evaluate the factors that can affect nail element concentrations for further sample analyses of a group of individuals by applying neutron activation analysis (NAA). Fingernails and toenails collected from adult individuals of both genders, aged 18 to 71 years, living in the Sao Paulo Metropolitan Region were cut in small fragments, cleaned and dried for analyses. Samples and element standards were irradiated for 16 h under a thermal neutron flux of about 4.5 x 10 12 n cm -2 s -1 at the IEA-R1 nuclear research reactor followed by gamma ray spectrometry. Element concentrations for As, Br, Ca, Co, Cr, Cs, Fe, K, La, Na, Rb, Sb, Sc, Se and Zn were determined. For quality control of the analytical results, certified reference materials were analysed and the results showed good accuracy and precision with relative errors and relative standard deviations lower than 5.1 % and 11.6 %, respectively. Preliminary assays indicated that the contribution due to impurities from plastic involucres used in the irradiation as well as those from nail polishes is very low and could be considered negligible. Results from the nail sample cleaning process using distinct procedures indicated that HNO 3 solution may cause sample dissolution. Sample homogeneity was verified by analysis of a sample in replicate. A

  4. Oxygen tension is a determinant of the matrix-forming phenotype of cultured human meniscal fibrochondrocytes.

    Directory of Open Access Journals (Sweden)

    Adetola B Adesida

    Full Text Available BACKGROUND: Meniscal cartilage displays a poor repair capacity, especially when injury is located in the avascular region of the tissue. Cell-based tissue engineering strategies to generate functional meniscus substitutes is a promising approach to treat meniscus injuries. Meniscus fibrochondrocytes (MFC can be used in this approach. However, MFC are unable to retain their phenotype when expanded in culture. In this study, we explored the effect of oxygen tension on MFC expansion and on their matrix-forming phenotype. METHODOLOGY/PRINCIPAL FINDINGS: MFC were isolated from human menisci followed by basic fibroblast growth factor (FGF-2 mediated cell expansion in monolayer culture under normoxia (21%O(2 or hypoxia (3%O(2. Normoxia and hypoxia expanded MFC were seeded on to a collagen scaffold. The MFC seeded scaffolds (constructs were cultured in a serum free chondrogenic medium for 3 weeks under normoxia and hypoxia. Constructs containing normoxia-expanded MFC were subsequently cultured under normoxia while those formed from hypoxia-expanded MFC were subsequently cultured under hypoxia. After 3 weeks of in vitro culture, the constructs were assessed biochemically, histologically and for gene expression via real-time reverse transcription-PCR assays. The results showed that constructs under normoxia produced a matrix with enhanced mRNA ratio (3.5-fold higher; p<0.001 of collagen type II to I. This was confirmed by enhanced deposition of collagen II using immuno-histochemistry. Furthermore, the constructs under hypoxia produced a matrix with higher mRNA ratio of aggrecan to versican (3.5-fold, p<0.05. However, both constructs had the same capacity to produce a glycosaminoglycan (GAG -specific extracellular matrix. CONCLUSIONS: Our data provide evidence that oxygen tension is a key player in determining the matrix phenotype of cultured MFC. These findings suggest that the use of normal and low oxygen tension during MFC expansion and subsequent neo

  5. A study on chemical element determinations in human nails by neutron activation analysis

    Energy Technology Data Exchange (ETDEWEB)

    Sanches, Thalita Pinheiro; Saiki, Mitiko, E-mail: thalitapsanches@usp.br, E-mail: mitiko@ipen.br [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

    2013-07-01

    Nail analyses have been the object of study in order to assess the levels of elements accumulated in the human organism and to use this tissue to monitor environmental and occupational exposure, to evaluate the nutritional status, to verify intoxication by toxic metals and to diagnose or to prevent diseases. Nail analyses present advantages due to easy sample collection, storage, transportation and this tissue provides element level accumulation over time. However, there is controversy regarding the application of nail analysis data due to difficulties to establish reliable reference values or element concentration ranges as control values. The objective of this study was to evaluate the factors that can affect nail element concentrations for further sample analyses of a group of individuals by applying neutron activation analysis (NAA). Fingernails and toenails collected from adult individuals of both genders, aged 18 to 71 years, living in the Sao Paulo Metropolitan Region were cut in small fragments, cleaned and dried for analyses. Samples and element standards were irradiated for 16 h under a thermal neutron flux of about 4.5 x 10{sup 12} n cm{sup -2} s{sup -1} at the IEA-R1 nuclear research reactor followed by gamma ray spectrometry. Element concentrations for As, Br, Ca, Co, Cr, Cs, Fe, K, La, Na, Rb, Sb, Sc, Se and Zn were determined. For quality control of the analytical results, certified reference materials were analysed and the results showed good accuracy and precision with relative errors and relative standard deviations lower than 5.1 % and 11.6 %, respectively. Preliminary assays indicated that the contribution due to impurities from plastic involucres used in the irradiation as well as those from nail polishes is very low and could be considered negligible. Results from the nail sample cleaning process using distinct procedures indicated that HNO{sub 3} solution may cause sample dissolution. Sample homogeneity was verified by analysis of a sample in

  6. Combined lipidomic and proteomic analysis of isolated human islets exposed to palmitate reveals time-dependent changes in insulin secretion and lipid metabolism.

    Directory of Open Access Journals (Sweden)

    Kirsten Roomp

    Full Text Available Studies on the pathophysiology of type 2 diabetes mellitus (T2DM have linked the accumulation of lipid metabolites to the development of beta-cell dysfunction and impaired insulin secretion. In most in vitro models of T2DM, rodent islets or beta-cell lines are used and typically focus is on specific cellular pathways or organs. Our aim was to, firstly, develop a combined lipidomics and proteomics approach for lipotoxicity in isolated human islets and, secondly, investigate if the approach could delineate novel and/ or confirm reported mechanisms of lipotoxicity. To this end isolated human pancreatic islets, exposed to chronically elevated palmitate concentrations for 0, 2 and 7 days, were functionally characterized and their levels of multiple targeted lipid and untargeted protein species determined. Glucose-stimulated insulin secretion from the islets increased on day 2 and decreased on day 7. At day 7 islet insulin content decreased and the proinsulin to insulin content ratio doubled. Amounts of cholesterol, stearic acid, C16 dihydroceramide and C24:1 sphingomyelin, obtained from the lipidomic screen, increased time-dependently in the palmitate-exposed islets. The proteomic screen identified matching changes in proteins involved in lipid biosynthesis indicating up-regulated cholesterol and lipid biosynthesis in the islets. Furthermore, proteins associated with immature secretory granules were decreased when palmitate exposure time was increased despite their high affinity for cholesterol. Proteins associated with mature secretory granules remained unchanged. Pathway analysis based on the protein and lipid expression profiles implicated autocrine effects of insulin in lipotoxicity. Taken together the study demonstrates that combining different omics approaches has potential in mapping of multiple simultaneous cellular events. However, it also shows that challenges exist for effectively combining lipidomics and proteomics in primary cells. Our

  7. Neutron activation analysis applied to the determination of trace elements in human nails

    International Nuclear Information System (INIS)

    Aguiar, Amilton Reinaldo

    2001-01-01

    There is a considerable interest in the determination of trace elements in human nails in order to use this tissue as a monitor of nutritional and healthy status of individuals, of occupational exposure diseases and of the environmental contamination. In this work, instrumental neutron activation analysis was applied to determine trace elements in finger nail clippings to make comparisons between the elemental concentrations obtained in nails from healthy individuals of a control group and those from cystic fibrosis (CF) patients. Firstly, a protocol for sample collection and preparation for analysis was established. Finger and toe nail samples from CF patients were collected at the Instituto da Crianca of Medicine School, University of Sao Paulo, SP. The nail samples from control group were collected from healthy adults and from children living in Sao Paulo city, SP. These samples were cleaned by stirring them witha a diluted Triton X100 solution and then by washing with distilled water and acetone. The analytical procedure consisted of irradiation nail samples and elemental standards in the IEA-R1m nuclear reactor under thermal neutro flux of about 10 12 n cm -2 s -1 , for short and long period irradiations. The activities of the radionuclides were measured using a gamma-ray spectrometer comprising an hyperpure Ge detector and associated electronic system. The biological reference materials Bovine Liver 1577b and Oyster Tissue 1566a, both from National Institute of Standards and Technology, USA were analysed in order to evaluate the accuracy and precision of the results. The analysis of variance (ANOVA) and F-test at the significance level of 5% were used to make a comparison between the sets of results obtained in this work. In the comparison of the results obtained for finger and toe nails from CF patients, the Br concentrations were higher in finger nails that those obtained for toe nails. For the control group of children, the finger nails presented different

  8. Sialo-Xenoantigenic Glycobiology Molecular Glycobiology of Sialylglycan-Xenoantigenic Determinants in Pig to Human Xenotransplantation

    CERN Document Server

    Song, Kwon-Ho

    2013-01-01

    Carbohydrate antigens on glycoconjugates of mammalian cells play crucial roles in various biological processes and are epitopes recognized by the immune system, as glycobiology has hugely been progressed during the past two decades. The book focuses on sialic acid–based xenoantigenes. In pig to human xenotransplantation, exposure of pig organs to human blood results in hyper acute rejection (HAR), caused by differences in carbohydrate epitopes between human and pig vascular endothelia. Although Gal-antigen as major antigen was eliminated, the remaining non-Gal antigens are considered to be xenoantigens. Sialosyl-Tn or Hanganutziu-Deicher (HD), are non-Gal antigens specific to natural antibodies in human. To overcome rejection responses such as HAR, studies of genes involved in carbohydrate antigens, causing xenoantigenicity, are necessary. Knowledge of pig glycosyltransferases are also useful to apply to xenoantigen masking or identification of the xenoantigenic sialylglycan(s). In the first chapter the scr...

  9. Determination of atomic number and composition of human enamel; Determinacao da composicao e numero atomico efetivo do esmalte humano

    Energy Technology Data Exchange (ETDEWEB)

    Nogueira, M.S. [Centro Regional de Ciencias Nucleares (CRCN), Recife, PE (Brazil); Rodas Duran, J.E. [Sao Paulo Univ., Ribeirao Preto, SP (Brazil). Faculdade de Filosofia, Ciencias e Letras. Dept. de Fisica e Matematica

    2001-07-01

    The teeth are organs of complicated structure that consist, partly, of hard tissue containing in its interior the dental pulp, rich in vases and nerves. The main mass of the tooth is constituted by the dentine, which is covered with hard tissues and of epithelial origin called enamel. The dentine of the human teeth used in this work were completely removed and the teeth were cut with a device with a diamond disc. In this work the chemical composition of the human enamel was determined, which showed a high percentage of Ca and P, in agreement with the results found in the literature. The effective atomic number of the material and the half-value layer in the energy range of diagnostic X-ray beams were determined. Teeth could be used to evaluated the public's individual doses as well as for retrospective dosimetry what confirms the importance of their effective atomic number and composition determination. (author)

  10. Understanding arsenic metabolism through spectroscopic determination of arsenic in human urine

    OpenAIRE

    Brima, Eid I.; Jenkins, Richard O.; Haris, Parvez I.

    2006-01-01

    In this review we discuss a range of spectroscopic techniques that are currently used for analysis of arsenic in human urine for understanding arsenic metabolism and toxicity, especially in relation to genetics/ethnicity, ingestion studies and exposure to arsenic through drinking water and diet. Spectroscopic techniques used for analysis of arsenic in human urine include inductively coupled plasma mass spectrometry (ICP-MS), hydride generation atomic absorption spectrometry (HG-AAS), hydride ...

  11. Determination of fluoride in human nails via cyclic instrumental neutron activation analysis

    International Nuclear Information System (INIS)

    Spate, V.L.; Morris, J.S.; Baskett, C.K.; Mason, M.M.; Reams, C.L.

    1994-01-01

    The role of fluorine in human health has become somewhat controversial. It is widely accepted as protective against dental caries, may be protective against osteoporosis, and has been very conservatively implicated with osteosarcoma in male rats. The develepment of a neutron activation analysis method and its application to the analysis of human nails is reported. It has been found that toenails collected in population-based epidemiology studies apparently reflect fluoride intake. (author) 11 refs.; 2 tabs

  12. Subcellular Distribution of NAD+ between Cytosol and Mitochondria Determines the Metabolic Profile of Human Cells*

    Science.gov (United States)

    VanLinden, Magali R.; Dölle, Christian; Pettersen, Ina K. N.; Kulikova, Veronika A.; Niere, Marc; Agrimi, Gennaro; Dyrstad, Sissel E.; Palmieri, Ferdinando; Nikiforov, Andrey A.; Tronstad, Karl Johan; Ziegler, Mathias

    2015-01-01

    The mitochondrial NAD pool is particularly important for the maintenance of vital cellular functions. Although at least in some fungi and plants, mitochondrial NAD is imported from the cytosol by carrier proteins, in mammals, the mechanism of how this organellar pool is generated has remained obscure. A transporter mediating NAD import into mammalian mitochondria has not been identified. In contrast, human recombinant NMNAT3 localizes to the mitochondrial matrix and is able to catalyze NAD+ biosynthesis in vitro. However, whether the endogenous NMNAT3 protein is functionally effective at generating NAD+ in mitochondria of intact human cells still remains to be demonstrated. To modulate mitochondrial NAD+ content, we have expressed plant and yeast mitochondrial NAD+ carriers in human cells and observed a profound increase in mitochondrial NAD+. None of the closest human homologs of these carriers had any detectable effect on mitochondrial NAD+ content. Surprisingly, constitutive redistribution of NAD+ from the cytosol to the mitochondria by stable expression of the Arabidopsis thaliana mitochondrial NAD+ transporter NDT2 in HEK293 cells resulted in dramatic growth retardation and a metabolic shift from oxidative phosphorylation to glycolysis, despite the elevated mitochondrial NAD+ levels. These results suggest that a mitochondrial NAD+ transporter, similar to the known one from A. thaliana, is likely absent and could even be harmful in human cells. We provide further support for the alternative possibility, namely intramitochondrial NAD+ synthesis, by demonstrating the presence of endogenous NMNAT3 in the mitochondria of human cells. PMID:26432643

  13. Subcellular Distribution of NAD+ between Cytosol and Mitochondria Determines the Metabolic Profile of Human Cells.

    Science.gov (United States)

    VanLinden, Magali R; Dölle, Christian; Pettersen, Ina K N; Kulikova, Veronika A; Niere, Marc; Agrimi, Gennaro; Dyrstad, Sissel E; Palmieri, Ferdinando; Nikiforov, Andrey A; Tronstad, Karl Johan; Ziegler, Mathias

    2015-11-13

    The mitochondrial NAD pool is particularly important for the maintenance of vital cellular functions. Although at least in some fungi and plants, mitochondrial NAD is imported from the cytosol by carrier proteins, in mammals, the mechanism of how this organellar pool is generated has remained obscure. A transporter mediating NAD import into mammalian mitochondria has not been identified. In contrast, human recombinant NMNAT3 localizes to the mitochondrial matrix and is able to catalyze NAD(+) biosynthesis in vitro. However, whether the endogenous NMNAT3 protein is functionally effective at generating NAD(+) in mitochondria of intact human cells still remains to be demonstrated. To modulate mitochondrial NAD(+) content, we have expressed plant and yeast mitochondrial NAD(+) carriers in human cells and observed a profound increase in mitochondrial NAD(+). None of the closest human homologs of these carriers had any detectable effect on mitochondrial NAD(+) content. Surprisingly, constitutive redistribution of NAD(+) from the cytosol to the mitochondria by stable expression of the Arabidopsis thaliana mitochondrial NAD(+) transporter NDT2 in HEK293 cells resulted in dramatic growth retardation and a metabolic shift from oxidative phosphorylation to glycolysis, despite the elevated mitochondrial NAD(+) levels. These results suggest that a mitochondrial NAD(+) transporter, similar to the known one from A. thaliana, is likely absent and could even be harmful in human cells. We provide further support for the alternative possibility, namely intramitochondrial NAD(+) synthesis, by demonstrating the presence of endogenous NMNAT3 in the mitochondria of human cells. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  14. Three-site sandwich radioimmunoassay with monoclonal antibodies for a sensitive determination of human alpha-fetoprotein

    International Nuclear Information System (INIS)

    Nomura, M.; Imai, M.; Takahashi, K.; Kumakura, T.; Tachibana, K.; Aoyagi, S.; Usuda, S.; Nakamura, T.; Miyakawa, Y.; Mayumi, M.

    1983-01-01

    Utilizing monoclonal antibodies against human alpha-fetoprotein, 3 distinct antigenic determinants were identified. These antigenic determinants, provisionally designated a, b and c, were arranged in such a manner that the binding of one determinant with the corresponding antibody did not inhibit, or only barely inhibited the binding of antibodies directed to the other 2 determinants. Monoclonal antibodies with 3 different specificities were, therefore, applied to develop a sandwich-type solid-phase radioimmunoassay of the antigen in which wells were coated with anti-a, and radiolabeled anti-b together with radiolabeled anti-c was employed to detect the bound antigen. The 3-site sandwich radioimmunoassay involving 3 different determinants gave a higher sensitivity than 2-site assays in which only anti-b or anti-c was employed as a radiolabeled reagent, because the radioactivity of the 2 labeled antibodies was added on the antigen bound to immobilized anti-a. (Auth.)

  15. Determination of the Optimized Automation Rate considering Effects of Automation on Human Operators in Nuclear Power Plants

    International Nuclear Information System (INIS)

    Lee, Seung Min; Seong, Poong Hyun; Kim, Jong Hyun; Kim, Man Cheol

    2015-01-01

    Automation refers to the use of a device or a system to perform a function previously performed by a human operator. It is introduced to reduce the human errors and to enhance the performance in various industrial fields, including the nuclear industry. However, these positive effects are not always achieved in complex systems such as nuclear power plants (NPPs). An excessive introduction of automation can generate new roles for human operators and change activities in unexpected ways. As more automation systems are accepted, the ability of human operators to detect automation failures and resume manual control is diminished. This disadvantage of automation is called the Out-of-the- Loop (OOTL) problem. We should consider the positive and negative effects of automation at the same time to determine the appropriate level of the introduction of automation. Thus, in this paper, we suggest an estimation method to consider the positive and negative effects of automation at the same time to determine the appropriate introduction of automation. This concept is limited in that it does not consider the effects of automation on human operators. Thus, a new estimation method for automation rate was suggested to overcome this problem

  16. Human and ecological determinants of the spatial structure of local breed diversity.

    Science.gov (United States)

    Colino-Rabanal, Victor J; Rodríguez-Díaz, Roberto; Blanco-Villegas, María José; Peris, Salvador J; Lizana, Miguel

    2018-04-24

    Since domestication, a large number of livestock breeds adapted to local conditions have been created by natural and artificial selection, representing one of the most powerful ways in which human groups have constructed niches to meet their need. Although many authors have described local breeds as the result of culturally and environmentally mediated processes, this study, located in mainland Spain, is the first aimed at identifying and quantifying the environmental and human contributions to the spatial structure of local breed diversity, which we refer to as livestock niche. We found that the more similar two provinces were in terms of human population, ecological characteristics, historical ties, and geographic distance, the more similar the composition of local breeds in their territories. Isolation by human population distance showed the strongest effect, followed by isolation by the environment, thus supporting the view of livestock niche as a socio-cultural product adapted to the local environment, in whose construction humans make good use of their ecological and cultural inheritances. These findings provide a useful framework to understand and to envisage the effects of climate change and globalization on local breeds and their livestock niches.

  17. Relation of circulating concentrations of chemokine receptor CCR5 ligands to C-peptide, proinsulin and HbA1c and disease progression in type 1 diabetes

    DEFF Research Database (Denmark)

    Pfleger, C; Kaas, A; Hansen, L

    2008-01-01

    Th1 related chemokines CCL3 and CCL5 and Th2 related CCL4 as ligands of the receptor CCR5 contribute to disease development in animal models of type 1 diabetes. In humans, no data are available addressing the role of these chemokines regarding disease progression and remission. We investigated lo...

  18. The Human and Physical Determinants of Wildfires and Burnt Areas in Israel

    Science.gov (United States)

    Levin, Noam; Tessler, Naama; Smith, Andrew; McAlpine, Clive

    2016-09-01

    Wildfires are expected to increase in Mediterranean landscapes as a result of climate change and changes in land-use practices. In order to advance our understanding of human and physical factors shaping spatial patterns of wildfires in the region, we compared two independently generated datasets of wildfires for Israel that cover approximately the same study period. We generated a site-based dataset containing the location of 10,879 wildfires (1991-2011), and compared it to a dataset of burnt areas derived from MODIS imagery (2000-2011). We hypothesized that the physical and human factors explaining the spatial distribution of burnt areas derived from remote sensing (mostly large fires, >100 ha) will differ from those explaining site-based wildfires recorded by national agencies (mostly small fires, human activities, improving the management of forest areas and raising public awareness to fire risk are key considerations in reducing fire danger.

  19. Kinetics of ingested 222Rn in humans determined from measurements with 133Xe. Project summary report

    International Nuclear Information System (INIS)

    Correia, J.A.; Weise, S.B.; Callahan, R.J.; Strauss, H.W.

    1987-12-01

    The problem of naturally occurring 222-radon contamination has received a great deal of public and scientific attention over the past several years, and has become a major public health issue worldwide. The purpose of the work reported in this document was to provide information about the behavior of ingested 222-radon in the digestive system and other organs of the human body. 133-Xenon, an element which behaves in the same manner as 222-radon in tissue and differs only in tissue solubility, was used in studies on human subjects. The tissue solubility differences were accounted for by using the tissue/blood partition coefficients of the two gases

  20. Closing the Gap: A Human Rights Approach towards Social Determinants of Health

    NARCIS (Netherlands)

    Toebes, Brigit; Stronks, Karien

    2016-01-01

    Social determinants of health are major contributors to population health as well as health inequalities. The public perception that health inequalities that arise from these social determinants are unjust seems to be widespread across societies. Nevertheless, there is also scepticism about the

  1. Closing the Gap : a Human Rights Approach to the Social Determinants of Health

    NARCIS (Netherlands)

    Toebes, Brigit; Stronks, Karien

    2016-01-01

    Social determinants of health are major contributors to population health as well as health inequalities. The public perception that health inequalities that arise from these social determinants are unjust seems to be widespread across societies. Nevertheless, there is also scepticism about the

  2. Definitions of the categories that determine the role of human in social and economic processes

    Directory of Open Access Journals (Sweden)

    Smachylo Valentina

    2016-01-01

    Full Text Available The priorities of the national economy development indicate the growing role of a person in the economic process of creating added value and capitalization of enterprises that require new approaches to the management process in this area. This requires the definition of basic categories that define the role and place of man in the socio-economic processes and characterise a person in the process of work. The article defines the basic aspects that must be considered in the study of the categories «staff», «personnel», «human resources», «cadre», «human potential», «cadre potential», «economically active population», «employment potential», «human capital»: evolution of concepts, level of socio-economic development, the presence or potentiality of human resources, the level of research, management paradigm. The essence, differentiation and interrelation of the given categories in the specified areas are justified. The necessity of socially responsible approach to management is underlined.

  3. Determination of markers for collagen type I turnover in peritendinous human tissue by microdialysis

    DEFF Research Database (Denmark)

    Olesen, J L; Langberg, Henning; Heinemeier, K M

    2006-01-01

    Previous results from our group have shown that loading of human tendon elevates tendinous type I collagen production measured by microdialysis. However, exclusion of the observed elevation as a response to trauma from inserting the microdialysis catheters or a possible influence from the collage...

  4. Determination Motive through the Prism of the General Concept of the Motives of Human Behaviour

    Science.gov (United States)

    Veresha, Roman V.

    2016-01-01

    This research studies the problems of defining the concept of motive of crime in terms of the psychological concept of motives of human behavior. The purpose of this research is to define the motive of crime (which is yet undefined in spite of the longstanding existence of criminology) and to improve existing scientific theories regarding the…

  5. Human Resource Development Practices as Determinant of HRD Climate and Quality Orientation

    Science.gov (United States)

    Hassan, Arif; Hashim, Junaidah; Ismail, Ahmad Zaki Hj

    2006-01-01

    Purpose: The aim of the study was to measure employees' perception of human resource development (HRD) practices, to explore whether ISO certification leads to any improvements in HRD system, and to examine the role of HRD practices on employees' development climate and quality orientation in the organization. Design/methodology/approach: A total…

  6. Human disturbance is a major determinant of wildlife distribution in Himalayan midhill landscapes of Nepal

    Czech Academy of Sciences Publication Activity Database

    Paudel, Prakash K.; Kindlmann, Pavel

    2012-01-01

    Roč. 15, č. 3 (2012), s. 283-293 ISSN 1367-9430 R&D Projects: GA MŠk(CZ) ED1.1.00/02.0073 Institutional support: RVO:67179843 Keywords : human disturbance * midhills * wildlife conservation * mountain landscape * conservation challenges Subject RIV: EH - Ecology, Behaviour Impact factor: 2.692, year: 2012

  7. Determination of platinum drug release and liposome stability in human plasma by CE-ICP-MS

    DEFF Research Database (Denmark)

    Nguyen, Trinh Thi Nhu Tam; Ostergaard, Jesper; Stürup, Stefan

    2013-01-01

    An in vitro method for simultaneous assessment of platinum release and liposome stability of liposomal formulations in human plasma is demonstrated. The development and assessment of the method was performed on a PEGylated liposomal formulation containing cisplatin. Complete separation of free ci...

  8. Aggrecan turnover in human intervertebral disc as determined by the racemization of aspartic acid

    NARCIS (Netherlands)

    Sivan, S.S.; Tsitron, E.; Wachtel, E.; Roughley, P.J.; Sakkee, N.; Ham, F. van der; Groot, J.de; Roberts, S.; Maroudas, A.

    2006-01-01

    We have used the racemization of aspartic acid as a marker for the "molecular age" of aggrecan components of the human intervertebral disc matrix (aggregating and non-aggregating proteoglycans as well as the different buoyant density fractions of aggrecan). By measuring the D/L Asp ratio of the

  9. Calcaneus length determines running economy: implications for endurance running performance in modern humans and Neandertals.

    Science.gov (United States)

    Raichlen, David A; Armstrong, Hunter; Lieberman, Daniel E

    2011-03-01

    The endurance running (ER) hypothesis suggests that distance running played an important role in the evolution of the genus Homo. Most researchers have focused on ER performance in modern humans, or on reconstructing ER performance in Homo erectus, however, few studies have examined ER capabilities in other members of the genus Homo. Here, we examine skeletal correlates of ER performance in modern humans in order to evaluate the energetics of running in Neandertals and early Homo sapiens. Recent research suggests that running economy (the energy cost of running at a given speed) is strongly related to the length of the Achilles tendon moment arm. Shorter moment arms allow for greater storage and release of elastic strain energy, reducing energy costs. Here, we show that a skeletal correlate of Achilles tendon moment arm length, the length of the calcaneal tuber, does not correlate with walking economy, but correlates significantly with running economy and explains a high proportion of the variance (80%) in cost between individuals. Neandertals had relatively longer calcaneal tubers than modern humans, which would have increased their energy costs of running. Calcaneal tuber lengths in early H. sapiens do not significantly differ from those of extant modern humans, suggesting Neandertal ER economy was reduced relative to contemporaneous anatomically modern humans. Endurance running is generally thought to be beneficial for gaining access to meat in hot environments, where hominins could have used pursuit hunting to run prey taxa into hyperthermia. We hypothesize that ER performance may have been reduced in Neandertals because they lived in cold climates. Copyright © 2011 Elsevier Ltd. All rights reserved.

  10. Determination of an unrelated donor pool size for human leukocyte antigen-matched platelets in Brazil

    Directory of Open Access Journals (Sweden)

    Carolina Bonet Bub

    2016-02-01

    Full Text Available ABSTRACT Background: Successful transfusion of platelet refractory patients is a challenge. Many potential donors are needed to sustain human leukocyte antigen matched-platelet transfusion programs because of the different types of antigens and the constant needs of these patients. For a highly mixed population such as the Brazilian population, the pool size required to provide adequate platelet support is unknown. Methods: A mathematical model was created to estimate the appropriate size of an unrelated donor pool to provide human leukocyte antigen-compatible platelet support for a Brazilian population. A group of 154 hematologic human leukocyte antigen-typed patients was used as the potential patient population and a database of 65,500 human leukocyte antigen-typed bone marrow registered donors was used as the donor population. Platelet compatibility was based on the grading system of Duquesnoy. Results: Using the mathematical model, a pool containing 31,940, 1710 and 321 donors would be necessary to match more than 80% of the patients with at least five completely compatible (no cross-reactive group, partial compatible (one cross-reactive group or less compatible (two cross-reactive group donors, respectively. Conclusion: The phenotypic diversity of the Brazilian population has probably made it more difficulty to find completely compatible donors. However, this heterogeneity seems to have facilitated finding donors when cross-reactive groups are accepted as proposed by the grading system of Duquesnoy. The results of this study may help to establish unrelated human leukocyte antigen-compatible platelet transfusions, a procedure not routinely performed in most Brazilian transfusion services.

  11. High Performace Liquid Chromtographic Determination of Nicardipine Hydrochloride in Human Plasma

    Directory of Open Access Journals (Sweden)

    Y. S. R. Krishnaiah

    2004-01-01

    Full Text Available A sensitive high-performance liquid chromatographic method was developed for the estimation of nicardipine hydrochloride in human plasma. Varying amount of nicardipine hydrochloride (2.5 to 150 ng/0.5 mL and fixed quantity (100 ng/0.5 mL of nifedipine (internal standard was added to blank human plasma, and a single step extraction was carried out with ethyl acetate. The mixture was centrifuged, ethyl acetate layer separated, dried and reconstituted with 100 μL of acetonitrile. Twenty microliters of this solution was injected into a reverse phase C-18 column using a mobile phase consisting of acetonitrile: 0.02 M potassium dihydrogen phosphate (pH 4.0 in the ratio of 60:40 v/v and the eluents were monitored at 239 nm. The method was validated for its linearity, precision and accuracy. The calibration curve was linear in the range of 5-150 ng/0.5 mL of plasma and the lower detection limit was 2.5 ng/0.5 mL of plasma. The intra- and inter-day variation was found to be less than 2.5% indicating that the method is highly precise. The mean recovery of nicardipine hydrochloride from plasma samples was 89.6±2.60%. The proposed HPLC method was applied for the estimation of nicardipine hydrochloride in human plasma after oral administration of an immediate release nicardipine hydrochloride capsule (dose 30 mg to 6 adult male volunteers. There was no interference of either the drug metabolites or other plasma components with the proposed HPLC method for the estimation of nicardipine hydrochloride in human plasma. Due to its simplicity, sensitivity, high precision and accuracy, the proposed HPLC method may be used for biopharmaceutical and pharmacokinetic evaluation of nicardipine hydrochloride and its formulations in humans

  12. Identification of the UBP1 locus as a critical blood pressure determinant using a combination of mouse and human genetics

    DEFF Research Database (Denmark)

    Koutnikova, Hana; Laakso, Markku; Lu, Lu

    2009-01-01

    complementarities of mouse and human genetic approaches, identifies the UBP1 locus as a critical blood pressure determinant. UBP1 plays a role in cholesterol and steroid metabolism via the transcriptional activation of CYP11A, the rate-limiting enzyme in pregnenolone and aldosterone biosynthesis. We suggest......Hypertension is a major health problem of largely unknown genetic origins. To identify new genes responsible for hypertension, genetic analysis of recombinant inbred strains of mice followed by human association studies might prove powerful and was exploited in our current study. Using a set of 27...... recombinant BXD strains of mice we identified a quantitative trait locus (QTL) for blood pressure (BP) on distal chromosome 9. The association analysis of markers encompassing the syntenic region on human chromosome 3 gave in an additive genetic model the strongest association for rs17030583 C/T and rs2291897...

  13. An investigation and comparison of speech recognition software for determining if bird song recordings contain legible human voices

    Directory of Open Access Journals (Sweden)

    Tim D. Hunt

    Full Text Available The purpose of this work was to test the effectiveness of using readily available speech recognition API services to determine if recordings of bird song had inadvertently recorded human voices. A mobile phone was used to record a human speaking at increasing distances from the phone in an outside setting with bird song occurring in the background. One of the services was trained with sample recordings and each service was compared for their ability to return recognized words. The services from Google and IBM performed similarly and the Microsoft service, that allowed training, performed slightly better. However, all three services failed to perform at a level that would enable recordings with recognizable human speech to be deleted in order to maintain full privacy protection.

  14. Determination of total and methylmercury compounds in the IAEA human hair intercomparison samples - Experience of the IAEA-MEL

    International Nuclear Information System (INIS)

    Horvat, M.; Liang, L.; Mandic, V.

    1995-01-01

    The programme of this CRP is focused on the analyses of human hair samples. There are only two human hair samples certified for total mercury, and no RMs for methylmercury compounds is available. One of the main objectives of this CRP is to produce, through the IAEA AQCS Programme, a human hair intercomparison material for quality assurance requirements in population monitoring programmes for total and methylmercury exposure. Through the reporting period, MESL has introduced a new method for simultaneous determination of total and methylmercury in biological samples. As the laboratory has close collaboration with the CRP's Reference Laboratory in Ljubljana, Slovenia, it has also been actively involved in the quality assurance component of this CRP. This report represents a summary on the results for total and methylmercury in two intercomparison samples, IAEA-085 and IAEA-086 using newly developed method

  15. Determination of γ-hydroxybutyrate in human urine samples by ion exclusion and ion exchange two-dimensional chromatography system.

    Science.gov (United States)

    Liu, Junwei; Deng, Zhifen; Zhu, Zuoyi; Wang, Yong; Wang, Guoqing; Sun, Yu-An; Zhu, Yan

    2017-12-15

    A two-dimensional ion chromatography system was developed for the determination of γ-hydroxybutyrate (GHB) in human urine samples. Ion exclusion chromatography was used in the first dimensional separation for elimination of urine matrices and detection of GHB above 10mgL -1 , ion exchange chromatography was used in the second dimensional separation via column-switching technique for detection of GHB above 0.08mgL -1 . Under the optimized chromatographic conditions, the ion exclusion and ion exchange chromatography separation system exhibited satisfactory repeatability (RSDchromatography system was convenient and practical for the determination of GHB in human urine samples. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. RP-HPLC Determination of Three Anti-Hyperlipidemic Drugs in Spiked Human Plasma and in Dosage Forms

    Directory of Open Access Journals (Sweden)

    Ola. M. Abdallah

    2011-01-01

    Full Text Available Sensitive, simple and accurate high performance liquid chromatographic (HPLC methods for the determination of atorvastatin (AT, fluvastatin (FL and pravastatin (PV have been developed. The proposed methods involve the use of a 150 mmx4.6 mm Zorbax Extend-C18 column (5 μm particle size and different chromatographic conditions for the separation of the three statins. Linearity range was 5-40, 5-30 and 10-60 μg mL-1 for AT, FL and PV respectively. The developed methods proved to be successful in the determination of all studied drugs in spiked human plasma samples.

  17. α-Klotho expression determines nitric oxide synthesis in response to FGF-23 in human aortic endothelial cells.

    Directory of Open Access Journals (Sweden)

    Chih-Ping Chung

    Full Text Available Endothelial cells (ECs express fibroblast growth factor (FGF receptors and are metabolically active after treatment with FGF-23. It is not known if this effect is α-Klotho independent or mediated by humoral or endogenous endothelial α-Klotho. In the present study, we aimed to characterize EC α-Klotho expression within the human vascular tree and to investigate the potential role of α-Klotho in determining FGF-23 mediated EC regulation. Human tissue and ECs from various organs were used for immunohistochemistry and Western blot. Primary cultures of human aortic endothelial cells (HAECs and human brain microvascular endothelial cells (HBMECs were used to generate in vitro cell models. We found endogenous α-Klotho expression in ECs from various organs except in microvascular ECs from human brain. Furthermore, FGF-23 stimulated endothelial nitric oxide synthase (eNOS expression, nitric oxide (NO production, and cell proliferation in HAECs. Interestingly, these effects were not observed in our HBMEC model in vitro. High phosphate treatment and endothelial α-Klotho knockdown mitigated FGF-23 mediated eNOS induction, NO production, and cell proliferation in HAECs. Rescue treatment with soluble α-Klotho did not reverse endothelial FGF-23 resistance caused by reduced or absent α-Klotho expression in HAECs. These novel observations provide evidence for differential α-Klotho functional expression in the human endothelium and its presence may play a role in determining the response to FGF-23 in the vascular tree. α-Klotho was not detected in cerebral microvascular ECs and its absence may render these cells nonresponsive to FGF-23.

  18. Simultaneous determination of triple therapy for Helicobacter pylori in human plasma by reversed phase chromatography with online wavelength switching

    Science.gov (United States)

    Ahmed, Sameh; Atia, Noha N.

    2015-02-01

    The infection of gastric mucosa by Helicobacter pylori (HP) is an essential cofactor in the aetiology of gastroduodenal ulcer and gastric carcinoma. Because of the bacterial resistance, combination therapy containing omeprazole (OME), tinidazole (TNZ) and clarithromycin (CLA) is commonly used for eradication of HP. However, the simultaneous determination of the triple therapy in human plasma was not reported. A simple, reproducible, and selective HPLC method was developed for the simultaneous determination of the triple therapy mixture used for management of HP infections in human plasma. An HPLC procedure based on a liquid-liquid extraction, enrichment of the analytes and subsequent reversed-phase chromatography with UV detection was used. To enable sensitive and selective detection, the method involved the use of online wavelength switching detection, with two different detection wavelengths; 280 nm for detection of OME and TNZ and 210 nm for detection of CLA. Separations were performed on C18 analytical column with acetonitrile-10 mM phosphate buffer of pH = 3.0 at flow rate of 1.0 mL min-1. The linear ranges in human plasma were 0.05-10 μg mL-1 with correlation coefficients >0.9990. The detection limits in human plasma were 0.02-0.07 μg mL-1. Validation parameters were assessed in compliance with US-FDA guidelines. The method proved to be valuable for the therapeutic drug monitoring after oral administration of triple therapy tablets.

  19. Determination of the distal dose edge in a human phantom by measuring the prompt gamma distribution: a Monte Carlo study

    Energy Technology Data Exchange (ETDEWEB)

    Min, Chul Hee; Lee, Han Rim; Yeom, Yeon Su; Cho, Sung Koo; Kim, Chan Hyeong [Hanyang University, Seoul (Korea, Republic of)

    2010-06-15

    The close relationship between the proton dose distribution and the distribution of prompt gammas generated by proton-induced nuclear interactions along the path of protons in a water phantom was demonstrated by means of both Monte Carlo simulations and limited experiments. In order to test the clinical applicability of the method for determining the distal dose edge in a human body, a human voxel model, constructed based on a body-composition-approximated physical phantom, was used, after which the MCNPX code was used to analyze the energy spectra and the prompt gamma yields from the major elements composing the human voxel model; finally, the prompt gamma distribution, generated from the voxel model and measured by using an array-type prompt gamma detection system, was calculated and compared with the proton dose distribution. According to the results, effective prompt gammas were produced mainly by oxygen, and the specific energy of the prompt gammas, allowing for selective measurement, was found to be 4.44 MeV. The results also show that the distal dose edge in the human phantom, despite the heterogeneous composition and the complicated shape, can be determined by measuring the prompt gamma distribution with an array-type detection system.

  20. Macrodistribution of 226Ra in the human skeleton as determined in vitro by gamma-ray spectrometry

    International Nuclear Information System (INIS)

    Essling, M.A.; Huff, D.R.

    1976-01-01

    A large number of γ-ray measurements of 226 Ra in human bone has been made both at MIT and CHR-ANL. These data are analyzed for correlations of the specific activity ratio of a bone group with other parameters, an interlaboratory comparison of the measurements, and an estimate of the errors expected when a whole-body content is calculated from the 226 Ra determination on a partial skeleton

  1. Determination of copper and iron in the human aqueous humor by atomic absorption spectrometer with graphite furnace

    International Nuclear Information System (INIS)

    Iqbal, Z.; Mohammad, Z.; Shah, M.T.; Saeed, M.; Imdadullah

    1999-01-01

    The concentration of copper and iron was determined in human aqueous humor using atomic absorption spectrophotometer equipped with graphite furnace. The mean (+- SEM) concentrations of copper (n=16) and iron (n=14) were 0.0234 -+ 0.0045 mu g.ml/sup -1/ and 0.045 -+ 0.0092 mu.ml/sup -1/ respectively. In male and female, the concentrations of copper (p< 0.82) and iron (p<0.38) were not significantly different. (author)

  2. Determination of gold accumulation in human tissues caused by gold therapy using x-ray fluorescence analysis

    International Nuclear Information System (INIS)

    Bacso, J.; Uzonyi, I.; Dezsoe, B.

    1986-08-01

    Human autopsy tissues from five patients with rheumatoid arthritis treated earlier with aqueous solution of gold and those from untreated control with the same disease were analyzed by x-ray fluorescence spectrometry using a conventional Si(Li) detection system. The gold and zinc concentrations of tissues were determined and compared with literature data. Correlation was found between Zn and Au concentrations in heart, lung, kidney and liver tissues. (author)

  3. Determination of seven trace elements in human milk, powdered cow's milk and infant foods by neutron activation analysis

    International Nuclear Information System (INIS)

    Grimanis, A.P.; Vassilaki-Grimani, M.; Alexiou, D.; Papadatos, C.

    1979-01-01

    The purpose of this study was to apply neutron activation analysis to the determination of seven trace elements (Co, Cr, Cu, Se, Zn, Rb and As) in colostrum, transitional and mature human milk as well as in powdered cow's milk and commercial infant foods, and thus to find out whether non-breast-fed infants received the same or different amounts of these trace elements as breast-fed ones. Ranges and averages for trace elements in mature human milk, expressed as microgrammes per gramme wet weight, varied as follows: Cr 0.018-0.040 and 0.027; Co 0.0013-0.0030 and 0.0020; Cu 0.38-0.50 and 0.46; Se 0.011-0.022 and 0.015; Zn 1.4-1.7 and 1.5; Rb 0.60-0.66 and 0.63; and As 0.0016-0.0060 and 0.0032. The mean concentrations of the essential trace elements Zn, Se, Cu, Cr and Co in colostrum were, respectively, 3.7, 3.2, 2.4, 1.8 and 1.7 times higher than those in mature human milk. No significant differences in Rb and As concentrations were found between colostrum and mature milk. Average values of Zn and Cu in transitional human milk were, respectively, 3.4 and 1.5 times higher than in mature milk. No significant differences for the rest of the elements were found between these two milk samples. The two milk formulas, half-cream and humanized, contained higher mean concentrations of Zn, Rb and As and similar or lower concentrations of Cr, Co and Se than mature human milk. Half-cream is deficient in Cu; mature human milk contains about 9 times more Cu than the half-cream cow's milk. Humanized milk contains a similar mean concentration of Cu as the mature milk. All examined infant foods contained similar or higher concentrations of all elements determined than did the mature human milk

  4. Effect of training on epinephrine-stimulated lipolysis determined by microdialysis in human adipose tissue

    DEFF Research Database (Denmark)

    Stallknecht, Bente; Simonsen, L; Bülow, J

    1995-01-01

    Trained humans (Tr) have a higher fat oxidation during submaximal physical work than sedentary humans (Sed). To investigate whether this reflects a higher adipose tissue lipolytic sensitivity to catecholamines, we infused epinephrine (0.3 nmol.kg-1.min-1) for 65 min in six athletes and six....... During epinephrine infusion intercellular glycerol concentrations were lower, but adipose tissue blood flow was higher in trained compared with sedentary subjects (P ... glycerol concentrations (Tr: 129 +/- 36 microM; Sed: 119 +/- 56) did not differ between groups. It is concluded that in intact subcutaneous adipose tissue epinephrine-stimulated blood flow is enhanced, whereas lipolytic sensitivity to epinephrine is the same in trained compared with untrained subjects....

  5. Effect of training on epinephrine-stimulated lipolysis determined by microdialysis in human adipose tissue

    DEFF Research Database (Denmark)

    Stallknecht, B; Simonsen, L; Bülow, J

    1995-01-01

    glycerol concentrations (Tr: 129 +/- 36 microM; Sed: 119 +/- 56) did not differ between groups. It is concluded that in intact subcutaneous adipose tissue epinephrine-stimulated blood flow is enhanced, whereas lipolytic sensitivity to epinephrine is the same in trained compared with untrained subjects.......Trained humans (Tr) have a higher fat oxidation during submaximal physical work than sedentary humans (Sed). To investigate whether this reflects a higher adipose tissue lipolytic sensitivity to catecholamines, we infused epinephrine (0.3 nmol.kg-1.min-1) for 65 min in six athletes and six...... sedentary young men. Glycerol was measured in arterial blood, and intercellular glycerol concentrations in abdominal subcutaneous adipose tissue were measured by microdialysis. Adipose tissue blood flow was measured by 133Xe-washout technique. From these measurements adipose tissue lipolysis was calculated...

  6. Design and characterization of a wearable macrobending fiber optic sensor for human joint angle determination

    Science.gov (United States)

    Silva, Ana S.; Catarino, André; Correia, Miguel V.; Frazão, Orlando

    2013-12-01

    The work presented here describes the development and characterization of intensity fiber optic sensor integrated in a specifically designed piece of garment to measure elbow flexion. The sensing head is based on macrobending incorporated in the garment, and the increase of curvature number was studied in order to investigate which scheme provided a good result in terms of sensitivity and repeatability. Results showed the configuration that assured a higher sensitivity (0.644 dBm/deg) and better repeatability was the one with four loops. Ultimately, this sensor can be used for rehabilitation purposes to monitor human joint angles, namely, elbow flexion on stroke survivors while performing the reach functional task, which is the most common upper-limb human gesture.

  7. The function of the human interferon-beta 1a glycan determined in vivo

    DEFF Research Database (Denmark)

    Dissing-Olesen, Lasse; Thaysen-Andersen, Morten; Meldgaard, Michael

    2008-01-01

    Recombinant human interferon-beta (rhIFN-beta) is the leading therapeutic intervention shown to change the cause of relapsing-remitting multiple sclerosis, and both a nonglycosylated and a significantly more active glycosylated variant of rhIFN-beta are used in treatment. This study investigates...... into the role of the rhIFN-beta1a glycan and its carbohydrate residues. The possibilities of improving the pharmacological properties of rhIFN-beta1a using glycoengineering are discussed...

  8. Application of thermoresponsive HPLC to forensic toxicology: determination of barbiturates in human urine

    OpenAIRE

    Kanno, Sanae; Watanabe, Kanako; Hirano, Seishiro; Yamagishi, Itaru; Gonmori, Kunio; Minakata, Kayoko; Suzuki, Osamu

    2009-01-01

    A high-performance liquid chromatography (HPLC) method has been developed for the assays of five barbiturates in human urine using a new thermoresponsive polymer separation column, which is composed of N-isopropylacrylamide polymer. According to elevating the column temperature from 10 ℃ to 50 ℃, five barbiturates, such as metharbital, primidone, phenobarbital, mephobarbital and pentobarbital, became well separated by this method. Five barbiturates showed good linearity in the range of 0.2-10...

  9. Age Effect in the Morphological Traits Performance for Sex Determination in Human Skulls and Mandibles

    OpenAIRE

    Suazo Galdames, Iván; Zavando, Daniela

    2012-01-01

    In this study we tested the hypothesis that diagnostic performance of the morphological indicators for sexual dimorphism are reduced as they are applied in skull and mandibles of older subjects. We used 275 adult human skulls, 250 of these with mandible, all subjects with sex and age registry. Sixteen classic morphological indicators of sexual dimorphism were evaluated, this information was compared with the registry and results noted in terms of precision. The best general performance of mor...

  10. The Acinar Cage: Basement Membranes Determine Molecule Exchange and Mechanical Stability of Human Breast Cell Acini.

    Directory of Open Access Journals (Sweden)

    Aljona Gaiko-Shcherbak

    Full Text Available The biophysical properties of the basement membrane that surrounds human breast glands are poorly understood, but are thought to be decisive for normal organ function and malignancy. Here, we characterize the breast gland basement membrane with a focus on molecule permeation and mechanical stability, both crucial for organ function. We used well-established and nature-mimicking MCF10A acini as 3D cell model for human breast glands, with ether low- or highly-developed basement membrane scaffolds. Semi-quantitative dextran tracer (3 to 40 kDa experiments allowed us to investigate the basement membrane scaffold as a molecule diffusion barrier in human breast acini in vitro. We demonstrated that molecule permeation correlated positively with macromolecule size and intriguingly also with basement membrane development state, revealing a pore size of at least 9 nm. Notably, an intact collagen IV mesh proved to be essential for this permeation function. Furthermore, we performed ultra-sensitive atomic force microscopy to quantify the response of native breast acini and of decellularized basement membrane shells against mechanical indentation. We found a clear correlation between increasing acinar force resistance and basement membrane formation stage. Most important native acini with highly-developed basement membranes as well as cell-free basement membrane shells could both withstand physiologically relevant loads (≤ 20 nN without loss of structural integrity. In contrast, low-developed basement membranes were significantly softer and more fragile. In conclusion, our study emphasizes the key role of the basement membrane as conductor of acinar molecule influx and mechanical stability of human breast glands, which are fundamental for normal organ function.

  11. New Targets for Zika Virus Determined by Human-Viral Interactomic: A Bioinformatics Approach

    Directory of Open Access Journals (Sweden)

    Eduardo Esteves

    2017-01-01

    Full Text Available Identifying ZIKV factors interfering with human host pathways represents a major challenge in understanding ZIKV tropism and pathogenesis. The integration of proteomic, gene expression and Protein-Protein Interactions (PPIs established between ZIKV and human host proteins predicted by the OralInt algorithm identified 1898 interactions with medium or high score (≥0.7. Targets implicated in vesicular traffic and docking were identified. New receptors involved in endocytosis pathways as ZIKV entry targets, using both clathrin-dependent (17 receptors and independent (10 receptors pathways, are described. New targets used by the ZIKV to undermine the host’s antiviral immune response are proposed based on predicted interactions established between the virus and host cell receptors and/or proteins with an effector or signaling role in the immune response such as IFN receptors and TLR. Complement and cytokines are proposed as extracellular potential interacting partners of the secreted form of NS1 ZIKV protein. Altogether, in this article, 18 new human targets for structural and nonstructural ZIKV proteins are proposed. These results are of great relevance for the understanding of viral pathogenesis and consequently the development of preventive (vaccines and therapeutic targets for ZIKV infection management.

  12. Human rights versus legal control over women's reproductive self-determination.

    Science.gov (United States)

    Uberoi, Diya; de Bruyn, Maria

    2013-06-14

    States have a duty under international human rights law to protect people's health. Nonetheless, while some health-related policies and laws protect basic human rights, others violate fundamental rights when they criminalize, prohibit, and restrict access to necessary health services. For example, laws and regulations related to protection of life from conception, contraception, actions of pregnant women, and abortion can harm women and place women and health care providers in jeopardy of legal penalization. Given the adverse consequences of punitive and restrictive laws related to pregnancy, advocates, civil society groups, human rights groups, and government institutions must work together to promote, protect, and fulfill women's fundamental reproductive rights. Copyright © 2013 Uberoi and de Bruyn. This is an open access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

  13. Magnetic graphene oxide as adsorbent for the determination of polycyclic aromatic hydrocarbon metabolites in human urine.

    Science.gov (United States)

    Zhu, Linli; Xu, Hui

    2014-09-01

    Detection of monohydroxy polycyclic aromatic hydrocarbons metabolites in urine is an advisable and valid method to assess human environmental exposure to polycyclic aromatic hydrocarbons. In this work, novel Fe3O4/graphene oxide composites were prepared and their application in the magnetic solid-phase extraction of monohydroxy polycyclic aromatic hydrocarbons in urine was investigated by coupling with liquid chromatography and mass spectrometry. In the hybrid material, superparamagnetic Fe3O4 nanoparticles provide fast separation to simplify the analytical process and graphene oxide provides a large functional surface for the adsorption. The prepared magnetic nanocomposites were characterized by X-ray diffraction, scanning electron microscopy, transmission electron microscopy, and vibrating sample magnetometry. The experimental conditions were optimized systematically. Under the optimal conditions, the recoveries of these compounds were in the range of 98.3-125.2%, the relative standard deviations ranged between 6.8 and 15.5%, and the limits of detection were in the range of 0.01-0.15 ng/mL. The simple, quick, and affordable method was successfully used in the analysis of human urinary monohydroxy polycyclic aromatic hydrocarbons in two different cities. The results indicated that the monohydroxy polycyclic aromatic hydrocarbons level in human urine can provide useful information for environmental exposure to polycyclic aromatic hydrocarbons. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Human capital of the child population and its selected determinants: Case study of Lviv city, Ukraine

    Directory of Open Access Journals (Sweden)

    Pantyley Viktoriya

    2017-09-01

    Full Text Available In conditions of modern economic and political destabilisation in Ukraine, human capital of the child population is considered as the most reliable indicator of socioeconomic development of the country and its regions. The primary goal of the study is the analysis of health status as the main element of human capital among the child population aged 0-14, and external and family environmental factors influencing this population group in the city of Lviv (in the light of available statistics and materials of the author’s own sociological research. Research results show a potential crisis of human capital among the child population in Lviv city. Results of own field research show a statistically important correlation between the willingness of parents to have more children and the following independent variables: number of members in the household, and the household’s income. The state of health among the child population showed a significant correlation between the following variables: household’s income, life satisfaction among parents, availability of healthcare in the place of residence, financial affordability of healthcare, physical activity among children, influence of noise and unpleasant scents.

  15. A truly human interface: Interacting face-to-face with someone whose words are determined by a computer program

    Directory of Open Access Journals (Sweden)

    Kevin eCorti

    2015-05-01

    Full Text Available We use speech shadowing to create situations wherein people converse in person with a human whose words are determined by a conversational agent computer program. Speech shadowing involves a person (the shadower repeating vocal stimuli originating from a separate communication source in real-time. Humans shadowing for conversational agent sources (e.g., chat bots become hybrid agents (echoborgs capable of face-to-face interlocution. We report three studies that investigated people’s experiences interacting with echoborgs and the extent to which echoborgs pass as autonomous humans. First, participants in a Turing Test spoke with a chat bot via either a text interface or an echoborg. Human shadowing did not improve the chat bot’s chance of passing but did increase interrogators’ ratings of how human-like the chat bot seemed. In our second study, participants had to decide whether their interlocutor produced words generated by a chat bot or simply pretended to be one. Compared to those who engaged a text interface, participants who engaged an echoborg were more likely to perceive their interlocutor as pretending to be a chat bot. In our third study, participants were naïve to the fact that their interlocutor produced words generated by a chat bot. Unlike those who engaged a text interface, the vast majority of participants who engaged an echoborg neither sensed nor suspected a robotic interaction. These findings have implications for android science, the Turing Test paradigm, and human-computer interaction. The human body, as the delivery mechanism of communication, fundamentally alters the social psychological dynamics of interactions with machine intelligence.

  16. Empirical Determination of Efficient Sensing Frequencies for Magnetometer-Based Continuous Human Contact Monitoring

    Directory of Open Access Journals (Sweden)

    Seungho Kuk

    2018-04-01

    Full Text Available The high linear correlation between the smartphone magnetometer readings in close proximity can be exploited for physical human contact detection, which could be useful for such applications as infectious disease contact tracing or social behavior monitoring. Alternative approaches using other capabilities in smartphones have aspects that do not fit well with the human contact detection. Using Wi-Fi or cellular fingerprints have larger localization errors than close human contact distances. Bluetooth beacons could reveal the identity of the transmitter, threatening the privacy of the user. Also, using sensors such as GPS does not work for indoor contacts. However, the magnetometer correlation check works best in human contact distances that matter in infectious disease transmissions or social interactions. The omni-present geomagnetism makes it work both indoors and outdoors, and the measured magnetometer values do not easily reveal the identity and the location of the smartphone. One issue with the magnetometer-based contact detection, however, is the energy consumption. Since the contacts can take place anytime, the magnetometer sensing and recording should be running continuously. Therefore, how we address the energy requirement for the extended and continuous operation can decide the viability of the whole idea. However, then, we note that almost all existing magnetometer-based applications such as indoor location and navigation have used high sensing frequencies, ranging from 10 Hz to 200 Hz. At these frequencies, we measure that the time to complete battery drain in a typical smartphone is shortened by three to twelve hours. The heavy toll raises the question as to whether the magnetometer-based contact detection can avoid such high sensing rates while not losing the contact detection accuracy. In order to answer the question, we conduct a measurement-based study using independently produced magnetometer traces from three different

  17. Empirical Determination of Efficient Sensing Frequencies for Magnetometer-Based Continuous Human Contact Monitoring.

    Science.gov (United States)

    Kuk, Seungho; Kim, Junha; Park, Yongtae; Kim, Hyogon

    2018-04-27

    The high linear correlation between the smartphone magnetometer readings in close proximity can be exploited for physical human contact detection, which could be useful for such applications as infectious disease contact tracing or social behavior monitoring. Alternative approaches using other capabilities in smartphones have aspects that do not fit well with the human contact detection. Using Wi-Fi or cellular fingerprints have larger localization errors than close human contact distances. Bluetooth beacons could reveal the identity of the transmitter, threatening the privacy of the user. Also, using sensors such as GPS does not work for indoor contacts. However, the magnetometer correlation check works best in human contact distances that matter in infectious disease transmissions or social interactions. The omni-present geomagnetism makes it work both indoors and outdoors, and the measured magnetometer values do not easily reveal the identity and the location of the smartphone. One issue with the magnetometer-based contact detection, however, is the energy consumption. Since the contacts can take place anytime, the magnetometer sensing and recording should be running continuously. Therefore, how we address the energy requirement for the extended and continuous operation can decide the viability of the whole idea. However, then, we note that almost all existing magnetometer-based applications such as indoor location and navigation have used high sensing frequencies, ranging from 10 Hz to 200 Hz. At these frequencies, we measure that the time to complete battery drain in a typical smartphone is shortened by three to twelve hours. The heavy toll raises the question as to whether the magnetometer-based contact detection can avoid such high sensing rates while not losing the contact detection accuracy. In order to answer the question, we conduct a measurement-based study using independently produced magnetometer traces from three different countries. Specifically, we

  18. Determination of cobalt in human biological liquids from electrothermal atomic absorption spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Dragan, Felicia [University of Oradea, Faculty of Medicine and Pharmacy, 29 N Jiga, 410028 Oradea (Romania); HIncu, Lucian [University of Medicine and Pharmacy ' Carol Davila' , Faculty of Pharmacy, 6 Traian Vuia, 020956 Bucuresti (Romania); Bratu, Ioan, E-mail: fdragan@uoradea.r [National Institute for Research and Development of Isotopic and Molecular Technologies, 65-103 Donath, 400293 Cluj-Napoca (Romania)

    2009-08-01

    Problems and possibilities of the determination of Co in serum and urine samples by electrothermal atomic absorption spectrometry (ETAAS) are described. Optimal instrumental parameters as well as a suitable atomizer, calibration procedure and hydrogen peroxide as modifier are proposed for direct ETAAS measurement of Co in serum and urine. The detection limit achieved was 0.1 {mu}g L{sup -1} for both matrices and relative standard deviations varied in the range 5-20% depending on the Co concentration in the sample. The validity of the method was verified by the analyses of standard reference materials. For serum samples with Co content lower than the detection limit, a separation and preconcentration procedure based on liquid/liquid extraction is suggested prior to determination of Co in the organic phase by ETAAS. This procedure permits determination of 0.02 {mu}g L{sup -1} Co in serum samples with a relative standard deviation of 10-18%.

  19. SELF-DETERMINATION AND POWER: A HUMAN-CENTRIC APPROACH TO THE INTERNATIONAL RELATIONS

    Directory of Open Access Journals (Sweden)

    Shameer Modongal

    2017-05-01

    Full Text Available This paper analyzes the importance of self-determination in international relations. It explains the puzzle why some small regions or group of people are ready to be small states by separating from big powerful states. The self-determination of a nation is preferable to the people than a military power of the large state. The military power of a state may not translate into a better life of people. So people’s empowerment cannot be treated the necessary outcome of state’s military power. When one group of people feel as marginalized in national policymaking and its implications, they show a secessionist tendency. This paper contends that people may prefer their ideology, identity and self-determination than the power of the state.

  20. An improved method to determine neuromuscular properties using force laws - From single muscle to applications in human movements.

    Science.gov (United States)

    Siebert, T; Sust, M; Thaller, S; Tilp, M; Wagner, H

    2007-04-01

    We evaluate an improved method for individually determining neuromuscular properties in vivo. The method is based on Hill's equation used as a force law combined with Newton's equation of motion. To ensure the range of validity of Hill's equation, we first perform detailed investigations on in vitro single muscles. The force-velocity relation determined with the model coincides well with results obtained by standard methods (r=.99) above 20% of the isometric force. In addition, the model-predicted force curves during work loop contractions very well agree with measurements (mean difference: 2-3%). Subsequently, we deduce theoretically under which conditions it is possible to combine several muscles of the human body to model muscles. This leads to a model equation for human leg extension movements containing parameters for the muscle properties and for the activation. To numerically determine these invariant neuromuscular properties we devise an experimental method based on concentric and isometric leg extensions. With this method we determine individual muscle parameters from experiments such that the simulated curves agree well with experiments (r=.99). A reliability test with 12 participants revealed correlations r=.72-.91 for the neuromuscular parameters (p<.01). Predictions of similar movements under different conditions show mean errors of about 5%. In addition, we present applications in sports practise and theory.

  1. Determination of ramipril in human plasma and its fragmentation by UPLC-Q-TOF-MS with positive electrospray ionization

    Directory of Open Access Journals (Sweden)

    Szpot Paweł

    2015-06-01

    Full Text Available This report presents the application of ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry with positive electrospray ionization, to determine ramipril in human plasma. First, the proteins in human plasma were precipitated using acetonitrile, then the supernatant was extracted by ethyl acetate at pH 3 and finally, the extract was analyzed using a UPLC-QTOF- MS system. The method was validated and the coefficient of determination (R2 was > 0.999, the lower limit of quantification (LLOQ was 0.5 ng mL-1. Precision, recovery and stability were determined for three different concentrations of ramipril. RSD for this method ranged from 3.3 to 8.6 %. The intra-day mean recovery was from 65.3 to 97.3 %. In addition, the fragmentation of ramipril was studied. Due to high resolution of the spectrometer, it was possible to measure fragment masses accurately and determine their molecular and chemical formulas with high accuracy.

  2. [Determination of lidocaine and its metabolites in human plasma by liquid chromatography in combination with tandem mass spectrometry].

    Science.gov (United States)

    Xiang, Jin; Zhang, Cheng; Yu, Qin; Liang, Mao-Zhi; Qin, Yong-Ping; Nan, Feng

    2010-07-01

    To establish a liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method for the determination of lidocaine (LDC) and its metabolites, monoethylglycinexylidide (MEGX) and glycinexylidide (GX), in human plasma. METHODS; The assay was conducted with an API 3000 HPLC-MS/MS system consisted of a Ultimate C18 column (50 x 4.6 mm, 5 microm). The mobile phase consisted of methanol: 5 mmol/ L ammonium acetate (50:50, pH was adjusted to 5.0 by formic acid) and the flow rate was set at 0.2 mL/min. The alkalinized sample was extracted with ethyl acetate. After evaporation of the organic layer, the residue was dissolved in mobile phase and the drug was determined by HPLC-MS/MS using electrospray ionization. The calibration curve was linear in a range from 15.625 to 2000 ng/mL for LDC. Linear calibration curves were obtained in the range of 1.5625 to 200 ng/mL for both for MEGX and GX. The limit of quantification for LDC, MEGX and GX was set at 15.625, 1.5625 and 1.5625 ng/mL. This method for the quantitative determination of lidocaine and its metabolites in human plasma is simple, rapid, sensitive and accurate. Therefore it can be used for the determination of lidocaine and its metabolites in clinical practice.

  3. Qualitative and quantitative determination of human biomarkers by laser photoacoustic spectroscopy methods

    Science.gov (United States)

    Popa, C.; Bratu, A. M.; Matei, C.; Cernat, R.; Popescu, A.; Dumitras, D. C.

    2011-07-01

    The hypothesis that blood, urine and other body fluids and tissues can be sampled and analyzed to produce clinical information for disease diagnosis or therapy monitoring is the basis of modern clinical diagnosis and medical practice. The analysis of breath air has major advantages because it is a non-invasive method, represents minimal risk to personnel collecting the samples and can be often sampled. Breath air samples from the human subjects were collected using aluminized bags from QuinTron and analyzed using the laser photoacoustic spectroscopy (LPAS) technique. LPAS is used to detect traces of ethylene in breath air resulting from lipid peroxidation in lung epithelium following the radiotherapy and also traces of ammonia from patients subjected to hemodialysis for treatment of renal failure. In the case of patients affected by cancer and treated by external radiotherapy, all measurements were done at 10P(14) CO2 laser line, where the ethylene absorption coefficient has the largest value (30.4 cm-1 atm-1), whereas for patients affected by renal failure and treated by standard dialysis, all measurements were performed at 9R(30) CO2 laser line, where the ammonia absorption coefficient has the maximum value of 57 cm-1 atm-1. The levels of ethylene and ammonia in exhaled air, from patients with cancer and renal failure, respectively, were measured and compared with breath air contents from healthy humans. Human gas biomarkers were measured at sub-ppb (parts per billion) concentration sensitivities. It has been demonstrated that LPAS technique will play an important role in the future of exhaled breath air analysis. The key attributes of this technique are sensitivity, selectivity, fast and real time response, as well as its simplicity.

  4. Simultaneous determination of cytosine arabinoside, daunorubicin and etoposide in human plasma

    DEFF Research Database (Denmark)

    Krogh-Madsen, Mikkel; Hansen, Steen Honoré; Honoré, Per Hartvig

    2010-01-01

    A method for simultaneous bioanalysis of the three cytotoxic drugs cytosine arabinoside, daunorubicin and etoposide in human plasma was developed and validated. A HPLC method with ultra-violet and fluorescence detection, preceded by mixed-mode cation-exchange solid phase extraction sample....... The overall precision (% relative standard deviation) was within 0.2-13.5% and the recovery ranged between 86.1% and 110.1% for the three drugs at all concentrations tested. Plasma samples were stable for at least two months when stored at -20 degrees C. The method was successfully applied to quantification...

  5. The quantitative determination of uranium in human hair by fission track measurements

    International Nuclear Information System (INIS)

    Wilson, D.J.; Bentley, K.W.

    1985-01-01

    Human hairs containing a uranium burden were placed in contact with a mica sheet as the recording matrix and irradiated in a thermal neutron flux. The fission fragment tracks penetrating the mica were etched and counted. Calculations have been made to show the losses due to the fission fragment range being less than the diameter of the hair and for the variation of track density with distance from the line of contact between the hair and the mica. Experimental data from 50 μm diameter hair and those derived by calculation were compared. (author)

  6. Use of radioimmunology determination of LH-RH in human urine

    International Nuclear Information System (INIS)

    Bourguignon, J.P.; Dourcy, C.; Franchimont, P.

    1976-01-01

    The existence of endogenous LHRH like immunoreactivity is shown in human urines after appropriate extraction, by radioimmunoassay of LHRH. In normaly cycling and menopausal women the quantities of endogenous hormone found in urines are greater after acid extraction than those found after extraction at pH 7. Furthermore, the increase observed by extraction in acidified methanol is directly correlated and proportional to the quantity of hormone assayable by extraction at pH 7. The hypothesis of urinary excretion of LHRH as a polymer of immunoreactive units is suggested by this study [fr

  7. Trace element determination in human bones using the neutron activation analysis method

    International Nuclear Information System (INIS)

    Kramarski, Sila; Saiki, Mitiko; Borelli, Aurelio; Batalha, Joao R.F.

    1997-01-01

    This work presents the results obtained in the analysis of rib bone samples from normal human individuals by applying instrumental neutron activation analysis. In these analyses, the elements Br, Cl, Fe, K, Mg, Na, Rb, Sr and Zn were found at the ppm level and the elements Ca and P at the level of percentage. The precision and the of the results were evaluated by using biological reference materials NIST SRM 1577a Bovine Liver, IAEA A-11 Milk powder, NIES CRM 9 Sargasso e NIES CRM 10A Rice Flour Unpolished. (author). 5 refs., 3 tabs

  8. Determination of fracture toughness of human permanent and primary enamel using an indentation microfracture method.

    Science.gov (United States)

    Hayashi-Sakai, Sachiko; Sakai, Jun; Sakamoto, Makoto; Endo, Hideaki

    2012-09-01

    The purpose of the present study was to examine the fracture toughness and Vickers microhardness number of permanent and primary human enamel using the indentation microfracture method. Crack resistance and a parameter indirectly related to fracture toughness were measured in 48 enamel specimens from 16 permanent teeth and 12 enamel specimens obtained from six primary teeth. The Vickers microhardness number of the middle portion was greater than the upper portion in primary enamel. The fracture toughness was highest in the middle portion of permanent enamel, because fracture toughness greatly depends upon microstructure. These findings suggest that primary teeth are not miniature permanent teeth but have specific and characteristic mechanical properties.

  9. Determination of persistent organohalogenated pollutants in human hair reference material (BCR 397): an interlaboratory study

    Energy Technology Data Exchange (ETDEWEB)

    Gill, Udai [Health Canada, Product Safety Laboratory, Ottawa, Ontario (Canada); Covaci, Adrian [University of Antwerp, Toxicological Center, Wilrijk (Belgium); Ryan, John Jake [Health Canada, Bureau of Chemical Safety, Health Products and Food Branch, Ottawa, Ontario (Canada); Emond, Andre [Health Canada, Environmental Research Division, First Nations and Inuit Health Branch, Ottawa, Ontario K1A0L2 (Canada)

    2004-12-01

    A human powdered hair material (BCR 397) was tested for its content in persistent organic pollutants (POPs), such as polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs) and polybrominated diphenyl ethers (PBDEs). Using different methods, three laboratories (two from Canada and one from Belgium) analysed a powdered hair sample to evaluate some methodologies and to obtain consensus values for selected POPs. Measured values for all PCB congeners and p,p'-DDE were within a relative standard deviation (RSD) of 15%. These first results contribute to the accuracy and precision for POPs quantification in hair and render test results more comparable among different laboratories. (orig.)

  10. Microscopic age determination of human skeletons including an unknown but calculable variable

    DEFF Research Database (Denmark)

    Wallin, Johan Albert; Tkocz, Izabella; Kristensen, Gustav

    1994-01-01

    estimation, which includes the covariance matrix of four single equation residuals, improves the accuracy of age determination. The standard deviation, however, of age prediction remains 12.58 years. An experimental split of the data was made in order to demonstrate that the use of subgroups gives a false...

  11. Relation between chloroguanide bioactivation to cycloguanil and the genetically determined metabolism of mephenytoin in humans.

    Science.gov (United States)

    Funck-Brentano, C; Bosco, O; Jacqz-Aigrain, E; Keundjian, A; Jaillon, P

    1992-05-01

    It has been suggested recently that the bioactivation of chloroguanide hydrochloride (proguanil) to its active antimalarial metabolite cycloguanil cosegregates with the genetically determined polymorphism of mephenytoin hydroxylation. We determined the chloroguanide to cycloguanil ratio in urine after oral administration of a single dose of 200 mg proguanil either alone or together with 100 mg racemic mephenytoin or 40 mg dextromethorphan in a randomized crossover study performed in 24 healthy subjects. The mephenytoin hydroxylation index was also determined after administration of 100 mg racemic mephenytoin either alone or together with 200 mg proguanil. Two subjects were poor metabolizers and one subject was an intermediate metabolizer of mephenytoin. These three subjects had chloroguanide to cycloguanil ratios of more than 50. The 21 subjects with the extensive metabolizer phenotype for mephenytoin hydroxylation had chloroguanide to cycloguanil ratios of less than 10. The chloroguanide to cycloguanil ratio was not significantly altered by mephenytoin or dextromethorphan coadministration. The trend toward a correlation between chloroguanide/cycloguanil ratio and log mephenytoin hydroxylation index did not reach statistical significance. Inclusion of the dextromethorphan metabolic ratio into the model did not improve the relationship. These findings confirm that the bioactivation of chloroguanide to cycloguanil cosegregates with the genetically determined activity of the CYP2C family. However, the chloroguanide to cycloguanil ratio and the mephenytoin hydroxylation index do not similarly reflect the variable activity of CYP2C.

  12. Crystallization and Structure Determination of the Human Estrogen Receptor by X-Ray Diffraction

    National Research Council Canada - National Science Library

    Harrison, Stephen

    1997-01-01

    ...) bound with 31 base-pairs of the 5S rRNA gene promoter has been determined at 3.1 A resolution. Individual zinc fingers are positioned differently in the major groove and across the minor groove of DNA to span the entire length of the duplex...

  13. Determinants of maternal pregnancy one-carbon metabolism and newborn human DNA methylation profiles

    NARCIS (Netherlands)

    N.H. van Mil (Nina); M.I. Bouwl-Both (Marieke I.); L. Stolk (Lisette); M.M.P.J. Verbiest (Michael); A. Hofman (Albert); V.W.V. Jaddoe (Vincent); F.C. Verhulst (Frank); P.H.C. Eilers (Paul); A.G. Uitterlinden (André); E.A.P. Steegers (Eric); H.W. Tiemeier (Henning); R.P.M. Steegers-Theunissen (Régine)

    2014-01-01

    textabstractMaternal one-carbon (1-C) metabolism provides methylgroups for fetal development and programing by DNA methylation as one of the underlying epigenetic mechanisms. We aimed to investigate maternal 1-C biomarkers, folic acid supplement use, and MTHFR C677T genotype as determinants of 1-C

  14. Determination of free insulin-like growth factor-I in human serum

    DEFF Research Database (Denmark)

    Frystyk, J.; Skjærbæk, C.; Ivarsen, P.

    2001-01-01

    Two fundamentally different methods are currently used for the determination of free insulin-like growth factor-I (IGF-I): ultrafiltration by centrifugation (UF) and direct immunoradiometric assay (IRMA). The aim was to evaluate a commercial IRMA (DSL, Webster, TX, USA) and to compare it with UF....

  15. Proteomic-Biostatistic Integrated Approach for Finding the Underlying Molecular Determinants of Hypertension in Human Plasma.

    Science.gov (United States)

    Gajjala, Prathibha R; Jankowski, Vera; Heinze, Georg; Bilo, Grzegorz; Zanchetti, Alberto; Noels, Heidi; Liehn, Elisa; Perco, Paul; Schulz, Anna; Delles, Christian; Kork, Felix; Biessen, Erik; Narkiewicz, Krzysztof; Kawecka-Jaszcz, Kalina; Floege, Juergen; Soranna, Davide; Zidek, Walter; Jankowski, Joachim

    2017-08-01

    Despite advancements in lowering blood pressure, the best approach to lower it remains controversial because of the lack of information on the molecular basis of hypertension. We, therefore, performed plasma proteomics of plasma from patients with hypertension to identify molecular determinants detectable in these subjects but not in controls and vice versa. Plasma samples from hypertensive subjects (cases; n=118) and controls (n=85) from the InGenious HyperCare cohort were used for this study and performed mass spectrometric analysis. Using biostatistical methods, plasma peptides specific for hypertension were identified, and a model was developed using least absolute shrinkage and selection operator logistic regression. The underlying peptides were identified and sequenced off-line using matrix-assisted laser desorption ionization orbitrap mass spectrometry. By comparison of the molecular composition of the plasma samples, 27 molecular determinants were identified differently expressed in cases from controls. Seventy percent of the molecular determinants selected were found to occur less likely in hypertensive patients. In cross-validation, the overall R 2 was 0.434, and the area under the curve was 0.891 with 95% confidence interval 0.8482 to 0.9349, P hypertensive patients were found to be -2.007±0.3568 and 3.383±0.2643, respectively, P hypertensives and normotensives. The identified molecular determinants may be the starting point for further studies to clarify the molecular causes of hypertension. © 2017 American Heart Association, Inc.

  16. A Review of the Extraction and Determination Methods of Thirteen Essential Vitamins to the Human Body: An Update from 2010.

    Science.gov (United States)

    Zhang, Yuan; Zhou, Wei-E; Yan, Jia-Qing; Liu, Min; Zhou, Yu; Shen, Xin; Ma, Ying-Lin; Feng, Xue-Song; Yang, Jun; Li, Guo-Hui

    2018-06-19

    Vitamins are a class of essential nutrients in the body; thus, they play important roles in human health. The chemicals are involved in many physiological functions and both their lack and excess can put health at risk. Therefore, the establishment of methods for monitoring vitamin concentrations in different matrices is necessary. In this review, an updated overview of the main pretreatments and determination methods that have been used since 2010 is given. Ultrasonic assisted extraction, liquid⁻liquid extraction, solid phase extraction and dispersive liquid⁻liquid microextraction are the most common pretreatment methods, while the determination methods involve chromatography methods, electrophoretic methods, microbiological assays, immunoassays, biosensors and several other methods. Different pretreatments and determination methods are discussed.

  17. A Review of the Extraction and Determination Methods of Thirteen Essential Vitamins to the Human Body: An Update from 2010

    Directory of Open Access Journals (Sweden)

    Yuan Zhang

    2018-06-01

    Full Text Available Vitamins are a class of essential nutrients in the body; thus, they play important roles in human health. The chemicals are involved in many physiological functions and both their lack and excess can put health at risk. Therefore, the establishment of methods for monitoring vitamin concentrations in different matrices is necessary. In this review, an updated overview of the main pretreatments and determination methods that have been used since 2010 is given. Ultrasonic assisted extraction, liquid–liquid extraction, solid phase extraction and dispersive liquid–liquid microextraction are the most common pretreatment methods, while the determination methods involve chromatography methods, electrophoretic methods, microbiological assays, immunoassays, biosensors and several other methods. Different pretreatments and determination methods are discussed.

  18. Selective changes in expression of HLA class I polymorphic determinants in human solid tumors

    International Nuclear Information System (INIS)

    Natali, P.G.; Nicotra, M.R.; Bigotti, A.; Venturo, I.; Giacomini, P.; Marcenaro, L.; Russo, C.

    1989-01-01

    Analysis of surgical biopsies with monoclonal antibodies (mAbs) to framework determinants of major histocompatibility complex class I antigens has shown that malignant transformation is frequently associated with a marked loss of these cell surface molecules. The present study sought to determine whether more selective losses of major histocompatibility complex class I expression occur. Multiple specimens from 13 different types of primary and metastatic tumors were tested utilizing mAb BB7.2, which recognizes a polymorphic HLA-A2 epitope. In each case, expression of HLA-A,B,C molecules was determined by testing with mAb W6/32 directed to a framework HLA class I determinant. The authors have found that in HLA-A2-positive patients, HLA-A2 products are not detectable or are reduced in their expression in 70-80% of endometrial, colorectal, mammary, and renal tumors; in 40-60% of soft-tissue, skin, ovary, urinary bladder, prostate, and stomach tumors; and in 25-30% of melanomas and lung carcinomas tested. All tumors expressed the framework HLA-A,B.C determinant. The HLA-A2 epitope recognized by mAb BB7.2 is located in a portion of the HLA-A2 molecule postulated to react with the T-cell receptor. The selective loss of an HLA class I polymorphic epitope shown in this study may explain the mechanism by which tumor cells escape both T-cell recognition and natural killer cell surveillance

  19. Relation of circulating concentrations of chemokine receptor CCR5 ligands to C-peptide, proinsulin and HbA1c and disease progression in type 1 diabetes

    DEFF Research Database (Denmark)

    Pfleger, C.; Kaas, A.; Hansen, L.

    2008-01-01

    Th1 related chemokines CCL3 and CCL5 and Th2 related CCL4 as ligands of the receptor CCR5 contribute to disease development in animal models of type 1 diabetes. In humans, no data are available addressing the role of these chemokines regarding disease progression and remission. We investigated...... longitudinally circulating concentrations of CCR5 ligands of 256 newly diagnosed patients with type 1 diabetes. CCR5 ligands were differentially associated with beta-cell function and clinical remission. CCL5 was decreased in remitters and positively associated with HbA1c suggestive of a Th1 associated...... of CCR5 by therapeutic agents such as maraviroc may provide a new therapeutic target to ameliorate disease progression in type 1 diabetes. (C) 2008 Elsevier Inc. All rights reserved Udgivelsesdato: 2008/7...

  20. Ultra-performance liquid chromatographic determination of tocopherols and retinol in human plasma.

    Science.gov (United States)

    Bell, Edward C; John, Mathew; Hughes, Rodney J; Pham, Thu

    2014-10-01

    A rapid, selective and sensitive ultra-performance liquid chromatography method has been developed for the detection and quantification of tocopherols and retinol in human plasma. Alpha-tocopherol, gamma-tocopherol and retinol are assayed using fluorescence detection. Excitation/emission wavelengths are 295/330 nm and 325/470 nm for the analysis of both tocopherols and retinol, respectively. Retinol acetate is employed as the internal standard. The reversed-phase method incorporates gradient elution with a mobile phase consisting of methanol and acetonitrile. Separation of vitamin compounds is achieved using a bridged ethyl hybrid C18 column. The retention times for retinol, retinol acetate, gamma-tocopherol and alpha-tocopherol are 1.6, 1.8, 3.9 and 4.3 min, respectively. The limits of quantification for retinol, gamma-tocopherol and alpha-tocopherol were 0.02, 0.02 and 0.1 µg/mL, respectively. The assay method is suitable for the analysis of tocopherols and retinol in human plasma. The method may be applied following the ingestion of foods fortified with these fat-soluble vitamins. © The Author [2013]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  1. Simultaneous determination of ethamsylate, tramadol and lidocaine in human urine by capillary electrophoresis with electrochemiluminescence detection.

    Science.gov (United States)

    Li, Jianguo; Ju, Huangxian

    2006-09-01

    Ethamsylate, tramadol and lidocaine, partly excreted by the kidney, are generally used as hemostatic, analgesic and local anesthetic in surgery. We developed a simple and sensitive method for their simultaneous monitoring in human urine based on CE coupled with electrochemiluminescence detection by end-column mode. Under optimized conditions the proposed method yielded linear ranges from 5.0 x 10(-8) to 5.0 x 10(-5), 1.0 x 10(-7) to 1.0 x 10(-4) and 1.0 x 10(-7) to 1.0 x 10(-4) M with LODs of 8.0 x 10(-9) M (36 amol), 1.6 x 10(-8) M (72 amol) and 1.0 x 10(-8) M (45 amol) (S/N = 3) for ethamsylate, tramadol and lidocaine, respectively. The RSD for their simultaneous detection at 1.0 x 10(-6) M was 2.1, 2.8 and 3.2% (n = 7), respectively. For practical application an extraction step with ethyl acetate at pH 11 was performed to eliminate the influence of the sample ionic strength. The recoveries of ethamsylate, tramadol and lidocaine at different levels in human urine were between 87 and 95%. This method was used for simultaneous detection of ethamsylate, tramadol and lidocaine in clinic urine samples from two medicated patients. It was valuable in clinical and biochemical laboratories for monitoring these drugs for various purposes.

  2. Prior Knowledge about Objects Determines Neural Color Representation in Human Visual Cortex.

    Science.gov (United States)

    Vandenbroucke, A R E; Fahrenfort, J J; Meuwese, J D I; Scholte, H S; Lamme, V A F

    2016-04-01

    To create subjective experience, our brain must translate physical stimulus input by incorporating prior knowledge and expectations. For example, we perceive color and not wavelength information, and this in part depends on our past experience with colored objects ( Hansen et al. 2006; Mitterer and de Ruiter 2008). Here, we investigated the influence of object knowledge on the neural substrates underlying subjective color vision. In a functional magnetic resonance imaging experiment, human subjects viewed a color that lay midway between red and green (ambiguous with respect to its distance from red and green) presented on either typical red (e.g., tomato), typical green (e.g., clover), or semantically meaningless (nonsense) objects. Using decoding techniques, we could predict whether subjects viewed the ambiguous color on typical red or typical green objects based on the neural response of veridical red and green. This shift of neural response for the ambiguous color did not occur for nonsense objects. The modulation of neural responses was observed in visual areas (V3, V4, VO1, lateral occipital complex) involved in color and object processing, as well as frontal areas. This demonstrates that object memory influences wavelength information relatively early in the human visual system to produce subjective color vision. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  3. Determination of the potential bioavailability of plant microRNAs using a simulated human digestion process.

    Science.gov (United States)

    Philip, Anna; Ferro, Valerie A; Tate, Rothwelle J

    2015-10-01

    The "dietary xenomiR hypothesis" proposes that microRNAs (miRNAs) in foodstuffs survive transit through the mammalian gastrointestinal tract and pass into cells intact to affect gene regulation. However, debate continues as to whether dietary intake poses a feasible route for such exogenous gene regulators. Understanding on miRNA levels during pretreatments of human diet is essential to test their bioavailability during digestion. This study makes the novel first use of an in vitro method to eliminate the inherent complexities and variability of in vivo approaches used to test this hypothesis. Plant miRNA levels in soybean and rice were measured during storage, processing, cooking, and early digestion using real-time PCR. We have demonstrated for the first time that storage, processing, and cooking does not abolish the plant miRNAs present in the foodstuffs. In addition, utilizing a simulated human digestion system revealed significant plant miRNA bioavailability after early stage digestion for 75 min. Attenuation of plant messenger RNA and synthetic miRNA was observed under these conditions. Even after an extensive pretreatment, plant-derived miRNA, delivered by typical dietary ingestion, has a robustness that could make them bioavailable for uptake during early digestion. The potential benefit of these regulatory molecules in pharma nutrition could be explored further. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Positioning of chromosomes in human spermatozoa is determined by ordered centromere arrangement.

    Directory of Open Access Journals (Sweden)

    Olga S Mudrak

    Full Text Available The intranuclear positioning of chromosomes (CHRs is a well-documented fact; however, mechanisms directing such ordering remain unclear. Unlike somatic cells, human spermatozoa contain distinct spatial markers and have asymmetric nuclei which make them a unique model for localizing CHR territories and matching peri-centromere domains. In this study, we established statistically preferential longitudinal and lateral positioning for eight CHRs. Both parameters demonstrated a correlation with the CHR gene densities but not with their sizes. Intranuclear non-random positioning of the CHRs was found to be driven by a specific linear order of centromeres physically interconnected in continuous arrays. In diploid spermatozoa, linear order of peri-centromeres was identical in two genome sets and essentially matched the arrangement established for haploid cells. We propose that the non-random longitudinal order of CHRs in human spermatozoa is generated during meiotic stages of spermatogenesis. The specific arrangement of sperm CHRs may serve as an epigenetic basis for differential transcription/replication and direct spatial CHR organization during early embryogenesis.

  5. Integrated approaches for determination of environmental and human risks of persistent toxic substances

    International Nuclear Information System (INIS)

    Blaha, L.; Cupr, P.; Dusek, L.; Hilscherova, K.; Holoubek, I.; Klanova, J.

    2008-01-01

    Substances that are persistent and bioaccumulative often posses toxic characteristics and cause adverse human health or environmental effects. Basic objective of the long-term research project INCHEMBIOL undertaken by the Centre RECETOX are the complex studies of interactions among chemical compounds present in environmental compartments and their biological effects and studies of the fate of mainly persistent chemical compounds in the environment, their effects on the environment and living organisms including human. Destiny in this concept consists of a summary of transport (from their input in the environment, transport within the environmental compartment, where they are discharged, transport among compartments and long-range transport in the environment) and transformation processes (abiotic and biotic transformations). It also includes study of distribution equilibriums, properties conditioning their environmental behaviour, study of the transformation processes and their products. This complex approach is a part of long-term research activities of the centre RECETOX. In the contribution methods used and results obtained in exploration of the causality among chemical (presence of chemical compounds in the environment) and biological (mechanisms of effects on the living organisms) are described.

  6. Variability of tissue mineral density can determine physiological creep of human vertebral cancellous bone.

    Science.gov (United States)

    Kim, Do-Gyoon; Shertok, Daniel; Ching Tee, Boon; Yeni, Yener N

    2011-06-03

    Creep is a time-dependent viscoelastic deformation observed under a constant prolonged load. It has been indicated that progressive vertebral deformation due to creep may increase the risk of vertebral fracture in the long-term. The objective of this study was to examine the relationships of creep with trabecular architecture and tissue mineral density (TMD) parameters in human vertebral cancellous bone at a physiological static strain level. Architecture and TMD parameters of cancellous bone were analyzed using microcomputerized tomography (micro-CT) in specimens cored out of human vertebrae. Then, creep and residual strains of the specimens were measured after a two-hour physiological compressive constant static loading and unloading cycle. Creep developed (3877 ± 2158 με) resulting in substantial levels of non-recoverable post-creep residual strain (1797 ± 1391 με). A strong positive linear correlation was found between creep and residual strain (r = 0.94, p creep rate. The TMD variability (GL(COV)) was the strongest correlate of creep rate (r = 0.79, p < 0.001). This result suggests that TMD variability may be a useful parameter for estimating the long-term deformation of a whole vertebral body. The results further suggest that the changes in TMD variability resulting from bone remodeling are of importance and may provide an insight into the understanding of the mechanisms underlying progressive failure of vertebral bodies and development of a clinical fracture. Copyright © 2011 Elsevier Ltd. All rights reserved.

  7. Development and validation of an HPLC method for simultaneous determination of trimethoprim and sulfamethoxazole in human plasma.

    Science.gov (United States)

    Sayar, Esin; Sahin, Selma; Cevheroglu, Semsettin; Hincal, A Atilla

    2010-09-01

    The combination of trimethoprim (TMP) and sulfamethoxazole (SMX) is used in the treatment of many common infections such as urinary, respiratory and gastrointestinal tract infections. The aim of this study was to determine TMP and SMX simultaneously in human plasma samples by high performance liquid chromatography (HPLC) using antipyrine as the internal standard. Separation of the compounds was achieved on a reverse-phase C8 column packed with 5 microm dimethyl octadecylsilyl bonded amorphous silica (4.6 mm x 250 mm) column using a mobile phase consisted of potassium hydrogen phosphate, acetonitrile, methanol and water adjusted to pH 6.2. The mobile phase was delivered at a flow rate of 1 mL min- and the effluent was monitored using Max plot technique at 25 derees C. Retention times were 5 min for TMP, 7 min for antipyrine and 9 min for SMX. Quantitation limits were 10 ng mL(-1) for TMP and 50 ng mL(-1) for SMX. Our findings indicated that the developed HPLC method was precise, accurate, specific and sensitive for simultaneous determination of TMP and SMX. Proposed HPLC method was successfully applied for the analysis of TMP and SMX in human plasma after oral administration of a co-trimoxazole tablet to human volunteers.

  8. Environmental variation of arsenic levels in human blood determined by neutron activation analysis

    DEFF Research Database (Denmark)

    Heydorn, Kaj

    1970-01-01

    Denmark. Arsenic was determined by neutron activation analysis with radiochemical separation and re-irradiation yield determination. The precision and accuracy of the results have been carefully evaluated in order to permit quantitative tests for the significance of the observed differences. The results......Arsenic levels in blood plasma and red cells from patients with Blackfoot disease, a peripheral arteriosclerosis endemic to a small area in Taiwan, were studied in relation to healthy individuals from the same and other parts of Taiwan and compared with arsenic levels in a control group from...... from Taiwan followed a logarithmic normal distribution, and no difference was found between Blackfoot patients and their healthy family members. However, their overall arsenic levels were higher than the Taiwan average, presumably because of arsenic in their drinking water. Much lower levels were found...

  9. The issue of HPLC determination of endogenous lipoic acid in human plasma.

    Science.gov (United States)

    Sechovcová, Soňa; Královcová, Pavla; Kanďár, Roman; Ventura, Karel

    2018-05-01

    Lipoic acid (LA) is used extensively as a therapeutic agent for the treatment of various diseases. Many methods have been reported for the determination of LA plasma levels and its metabolites after its supplementation, but available information concerning endogenous plasma levels is still scarce. Studies which directly focused on determining the endogenous plasma levels provided highly controversial results, endogenous plasma levels of LA: 2.4 and 4.9 nmol/L respectively. However, the levels of free LA in the plasma of nonsupplemented voluntary blood donors were not detectable in all cases. The presented results of our study show that endogenous concentrations of LA are <1.85 nmol/L. Copyright © 2017 John Wiley & Sons, Ltd.

  10. Determination of bacterial endotoxins in blood of animals and humans subjected to irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Kiselev, P N; Shchul' s, T S [Tsentral' nyj Nauchno-Issledovatel' skij Rentgeno-Radiologicheskij Inst., Leningrad (USSR)

    1981-01-01

    The content of Gram-negative bacterium endotoxins in the irradiated organism was determined by the use of actinomycin D sensitizing the organism to the effect of such toxins. Regularities in the accumulation of Gram-negative bacterium endotoxins in the irradiated organism were studied as well as the dependence of animal sensitization on the actinomycin D dose. Possibilities for the development of endotoxemia in oncologic patients in the process of radiotheraphy were traced. It is shown that the radiotherapy of the intestine containing large quantaties of bacteria and their toxins is accompanied by endotoxemia; in the first part of the radiotherapy course its manifestation depends on the irradiation dose; having reached a specific level, endotoxemia retains its symptoms till the end of irradiation. It is indicated that the determination of bacterium endotoxins is of interest when studying not only the development of endotoxemia in the organism exposed to radiation, hut also many other toxemic states.

  11. Determining picogram quantities of U in human urine by thermal ionization mass spectrometry

    International Nuclear Information System (INIS)

    Kelly, W.R.; Fassett, J.D.; Hotes, S.A.

    1987-01-01

    The U concentration in Standard Reference Material 2670 (Toxic Metals in Freeze-Dried Urine) and the urine of two preschool-age children were determined by measuring the chemically separated U by isotope dilution thermal ionization mass spectrometry using ion counting detection. This procedure can detect about 1% of the U atoms loaded into the mass spectrometer and has a total chemical blank of about 5 pg U. The U concentration in SRM 2670 was found to be 113 +/- 2 pg 238 U/ml (1 s). At this concentration, a 1-ml sample is sufficient for a determination with a total uncertainty of less than 5%. The U concentrations in the two children were 3.1 +/- 0.9 and 3.6 +/- 0.9 pg 238 U/g. These values suggest that the U concentration in urine of unexposed persons may be at this low level or lower

  12. Determining the Neural Substrate for Encoding a Memory of Human Pain and the Influence of Anxiety.

    Science.gov (United States)

    Tseng, Ming-Tsung; Kong, Yazhuo; Eippert, Falk; Tracey, Irene

    2017-12-06

    To convert a painful stimulus into a briefly maintainable construct when the painful stimulus is no longer accessible is essential to guide human behavior and avoid dangerous situations. Because of the aversive nature of pain, this encoding process might be influenced by emotional aspects and could thus vary across individuals, but we have yet to understand both the basic underlying neural mechanisms as well as potential interindividual differences. Using fMRI in combination with a delayed-discrimination task in healthy volunteers of both sexes, we discovered that brain regions involved in this working memory encoding process were dissociable according to whether the to-be-remembered stimulus was painful or not, with the medial thalamus and the rostral anterior cingulate cortex encoding painful and the primary somatosensory cortex encoding nonpainful stimuli. Encoding of painful stimuli furthermore significantly enhanced functional connectivity between the thalamus and medial prefrontal cortex (mPFC). With regards to emotional aspects influencing encoding processes, we observed that more anxious participants showed significant performance advantages when encoding painful stimuli. Importantly, only during the encoding of pain, the interindividual differences in anxiety were associated with the strength of coupling between medial thalamus and mPFC, which was furthermore related to activity in the amygdala. These results indicate not only that there is a distinct signature for the encoding of a painful experience in humans, but also that this encoding process involves a strong affective component. SIGNIFICANCE STATEMENT To convert the sensation of pain into a briefly maintainable construct is essential to guide human behavior and avoid dangerous situations. Although this working memory encoding process is implicitly contained in the majority of studies, the underlying neural mechanisms remain unclear. Using fMRI in a delayed-discrimination task, we found that the

  13. Rapid determination of human globin chains using reversed-phase high-performance liquid chromatography.

    Science.gov (United States)

    Wan, Jun-Hui; Tian, Pei-Ling; Luo, Wei-Hao; Wu, Bing-Yi; Xiong, Fu; Zhou, Wan-Jun; Wei, Xiang-Cai; Xu, Xiang-Min

    2012-07-15

    Reversed-phase high-performance liquid chromatography (RP-HPLC) of human globin chains is an important tool for detecting thalassemias and hemoglobin variants. The challenges of this method that limit its clinical application are a long analytical time and complex sample preparation. The aim of this study was to establish a simple, rapid and high-resolution RP-HPLC method for the separation of globin chains in human blood. Red blood cells from newborns and adults were diluted in deionized water and injected directly onto a micro-jupiter C18 reversed-phase column (250 mm × 4.6 mm) with UV detection at 280 nm. Under the conditions of varying pH or the HPLC gradient, the globin chains (pre-β, β, δ, α, (G)γ and (A)γ) were denatured and separated from the heme groups in 12 min with a retention time coefficient of variation (CV) ranging from 0.11 to 1.29% and a peak area CV between 0.32% and 4.86%. Significant differences (P<0.05) among three groups (normal, Hb H and β thalassemia) were found in the area ratio of α/pre-β+β applying the rapid elution procedure, while P≥0.05 was obtained between the normal and α thalassemia silent/trait group. Based on the ANOVA results, receiver operating characteristic (ROC) curve analysis of the δ/β and α/pre-β+β area ratios showed a sensitivity of 100.0%, and a specificity of 100.0% for indicating β thalassemia carriers, and a sensitivity of 96.6% and a specificity of 89.6% for the prediction of hemoglobin H (Hb H) disease. The proposed cut-off was 0.026 of δ/β for β thalassemia carriers and 0.626 of α/pre-β+β for Hb H disease. In addition, abnormal hemoglobin hemoglobin E (Hb E) and Hb Westmead (Hb WS) were successfully identified using this RP-HPLC method. Our experience in developing this RP-HPLC method for the rapid separation of human globin chains could be of use for similar work. Copyright © 2012 Elsevier B.V. All rights reserved.

  14. Determination of Drug Toxicity Using 3D Spheroids Constructed From an Immortal Human Hepatocyte Cell Line

    DEFF Research Database (Denmark)

    Fey, S. J.; Wrzesinski, Krzysztof

    2012-01-01

    that a precise dose can be provided in a manner similar to in vivo studies. This avoided correction of the actual dose given based on a protein determination after treatment (when some cells may have lysed). Conversion of published in vitro LC50 data (mM) for six common drugs (acetaminophen, amiodarone...... different from 2D cultures and are more representative of the liver in vivo....

  15. Determination of Smuggled Cigarette Characteristics in Brazil and Their Potential Risk to the Human Health

    Directory of Open Access Journals (Sweden)

    Cleber Pinto da Silva

    2016-07-01

    Full Text Available The objective of this study was to determine the different characteristics of tobacco found in thirty brands of smuggled cigarettes in Brazil. Determination of arsenic through atomic absorption spectrometry in graphite oven was carried out and classical methodologies were employed to determine dirtiness, total ash, insoluble ash, humidity, tobacco pH and sidestream smoke pH. The methodology used to quantify arsenic presented quantification limit of 15.0 ng g-1 and detection limit of 4.0 ng g-1 in dry tobacco mass. The recovery of arsenic for the method purpose was 98.2% and relative standard deviation 6.0%. About 56% of the brands were observed to have arsenic concentrations above 20.0 ng g-1, which means nearly twice as much as the arsenic found in cigarettes sold legally in Brazil. Levels above the recommended value for humidity were found in 53% of brands. About 96% of the brands presented total ash content above that indicated by the Brazilian Pharmacopoeia. About 53% of the samples contained levels of insoluble ash above the limit. In 90% of the samples, the smoke was alkaline. In dirtiness tests, 81.2% of the brands presented some kind of contaminant, such as fungi, insect fragments, grass or mites. The characteristics revealed that the consumption of this kind of cigarette can increase risks to consumer health. DOI: http://dx.doi.org/10.17807/orbital.v8i3.832 

  16. Metric Sex Determination of the Human Coxal Bone on a Virtual Sample using Decision Trees.

    Science.gov (United States)

    Savall, Frédéric; Faruch-Bilfeld, Marie; Dedouit, Fabrice; Sans, Nicolas; Rousseau, Hervé; Rougé, Daniel; Telmon, Norbert

    2015-11-01

    Decision trees provide an alternative to multivariate discriminant analysis, which is still the most commonly used in anthropometric studies. Our study analyzed the metric characterization of a recent virtual sample of 113 coxal bones using decision trees for sex determination. From 17 osteometric type I landmarks, a dataset was built with five classic distances traditionally reported in the literature and six new distances selected using the two-step ratio method. A ten-fold cross-validation was performed, and a decision tree was established on two subsamples (training and test sets). The decision tree established on the training set included three nodes and its application to the test set correctly classified 92% of individuals. This percentage was similar to the data of the literature. The usefulness of decision trees has been demonstrated in numerous fields. They have been already used in sex determination, body mass prediction, and ancestry estimation. This study shows another use of decision trees enabling simple and accurate sex determination. © 2015 American Academy of Forensic Sciences.

  17. Health as freedom: addressing social determinants of global health inequities through the human right to development.

    Science.gov (United States)

    Fox, Ashley M; Meier, Benjamin Mason

    2009-02-01

    In spite of vast global improvements in living standards, health, and well-being, the persistence of absolute poverty and its attendant maladies remains an unsettling fact of life for billions around the world and constitutes the primary cause for the failure of developing states to improve the health of their peoples. While economic development in developing countries is necessary to provide for underlying determinants of health--most prominently, poverty reduction and the building of comprehensive primary health systems--inequalities in power within the international economic order and the spread of neoliberal development policy limit the ability of developing states to develop economically and realize public goods for health. With neoliberal development policies impacting entire societies, the collective right to development, as compared with an individual rights-based approach to development, offers a framework by which to restructure this system to realize social determinants of health. The right to development, working through a vector of rights, can address social determinants of health, obligating states and the international community to support public health systems while reducing inequities in health through poverty-reducing economic growth. At an international level, where the ability of states to develop economically and to realize public goods through public health systems is constrained by international financial institutions, the implementation of the right to development enables a restructuring of international institutions and foreign-aid programs, allowing states to enter development debates with a right to cooperation from other states, not simply a cry for charity.

  18. Hyperspectral imaging based on compressive sensing to determine cancer margins in human pancreatic tissue ex vivo

    Science.gov (United States)

    Peller, Joseph; Thompson, Kyle J.; Siddiqui, Imran; Martinie, John; Iannitti, David A.; Trammell, Susan R.

    2017-02-01

    Pancreatic cancer is the fourth leading cause of cancer death in the US. Currently, surgery is the only treatment that offers a chance of cure, however, accurately identifying tumor margins in real-time is difficult. Research has demonstrated that optical spectroscopy can be used to distinguish between healthy and diseased tissue. The design of a single-pixel imaging system for cancer detection is discussed. The system differentiates between healthy and diseased tissue based on differences in the optical reflectance spectra of these regions. In this study, pancreatic tissue samples from 6 patients undergoing Whipple procedures are imaged with the system (total number of tissue sample imaged was N=11). Regions of healthy and unhealthy tissue are determined based on SAM analysis of these spectral images. Hyperspectral imaging results are then compared to white light imaging and histological analysis. Cancerous regions were clearly visible in the hyperspectral images. Margins determined via spectral imaging were in good agreement with margins identified by histology, indicating that hyperspectral imaging system can differentiate between healthy and diseased tissue. After imaging the system was able to detect cancerous regions with a sensitivity of 74.50±5.89% and a specificity of 75.53±10.81%. Possible applications of this imaging system include determination of tumor margins during surgery/biopsy and assistance with cancer diagnosis and staging.

  19. Determination of total mercury and methylmercury in human head hair by radiochemical methods of analysis

    International Nuclear Information System (INIS)

    Vasconcellos, M.B.A.; Saiki, M.; Paletti, G.; Baruzzi, R.G.; Rodrigues, D.A.; Cuten, J.

    1995-01-01

    Total mercury has been determined by instrumental neutron activation analysis in the hair of several Indian tribes living in the Xingu Park, located in the Amazonic region of Brazil. Methylmercury and total mercury have been determined in selected samples using cold vapour atomic absorption spectroscopy, at the Nuclear Chemistry Department, Jozef Stefan Institute, Ljubliana, Slovenia. Mercury levels were found to be much higher in the Indian hair samples as compared to the samples from the control population. The arithmetic and geometric means for total mercury in the Indian hair samples ranged from 10 to 20 ppm, compared to values of about 1 ppm for the means of the control group. The results obtained for methylmercury have shown that the majority of the mercury is present in the hair of the Indians as the organic form. The Indian study populations living in the Xingu Park can thus be considered as being at risk with regards to contamination by mercury. With the aim of applying neutron activation analysis for the determination of methylmercury in hair, experiments were done at the IEA-R1 nuclear research reactor irradiating cysteine- and also thioacetamide- impregnated filter papers, on which a methylmercury solution was pipetted. The results obtained have shown that all the mercury was lost from the cysteine-impregnated paper and about 90 % of the mercury remained on the paper impregnated with thioacetamide. (author)

  20. Possibility of determination of the level of antioxidants in human body using spectroscopic methods

    International Nuclear Information System (INIS)

    Timofeeva, E; Gorbunova, E

    2016-01-01

    In this work, the processes of antioxidant defence against aggressive free radicals in human body were investigated theoretically; and the existing methods of diagnosis of oxidative stress and disturbance of antioxidant activity were reviewed. Also, the kinetics of free radical reactions in the oxidation of luminol and interaction antioxidants (such as chlorophyll in the multicomponent system of plant's leaves and ubiquinone) with the UV radiation were investigated experimentally by spectroscopic method. The results showed that this method is effective for recording the luminescence of antioxidants, free radicals, chemiluminescent reactions and fluorescence. In addition these results reveal new opportunities for the study of the antioxidant activity and antioxidant balance in a multicomponent system by allocating features of the individual components in spectral composition. A creation of quality control method for drugs, that are required for oxidative stress diagnosis, is a promising direction in the development of given work. (paper)

  1. Experiments on the radioimmunological determination of the triiodothyronine binding capacity of human blood serum

    International Nuclear Information System (INIS)

    Traub, H.W.J.

    1980-01-01

    Bovine serum antibodies have been obtained from rabbits. Constant amounts of antibodies have been incubated with increasing amounts of antigens. The maximum value found for T 3 binding to the antigen-antibody complex was 66 μg RA/mg antigen. Human serum samples containing 0,05 ng 125 J-T 3 /μl were incubated with 600 μg RA, and the antigen was precipitated with 8,4 mg anti-RA. The mean value obtained for 30 enthyreotic test persons was a binding to RA of 7.9 +- 0,52% 125 J-T 3 . Comparative measurements of patient and standard serum samples shared only slight differences. This means that the measured data do not reflect the thyroid function. (orig./MG) [de

  2. Mini-review: Can non-human leucocyte antigen genes determine susceptibility to severe dengue syndromes?

    Science.gov (United States)

    Ng, Dorothy; Ghosh, Aparna; Jit, Mark; Seneviratne, Suranjith L

    2017-09-01

    Dengue viral infections are endemic or epidemic in virtually all tropical countries. Among individuals infected with the dengue virus, severe dengue syndromes (i.e., dengue haemorrhagic fever and dengue shock syndromes) tend to affect only some and this may be due to a combination of host genetic susceptibility and viral factors. In this review article we analyse and discuss the present knowledge of non-human leucocyte antigen host genetic susceptibility to severe dengue syndromes. The relevance of genetic polymorphisms in the pathways of antigen recognition, uptake, processing and presentation, activation of interferon α responses, mast cell and complement activation and T cell activation and dengue disease severity has been reviewed and analysed. © The Author(s) 2018. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  3. A New Validated RP- HPLC Method for the Determination of Nevirapine in Human Plasma

    Directory of Open Access Journals (Sweden)

    C. H. Venkata Kumar

    2010-01-01

    Full Text Available A rapid, selective and sensitive high performance liquid chromatographic method for the estimation of nevirapine in human plasma has been developed. Chromatography was carried out on a Hypersil BDS C18 column using a mixture of ammonium acetate buffer (pH 4.0 ± 0.05 and acetonitrile (85:15 v/v as the mobile phase. The eluents were monitored for the drug by UV detection at 254 nm. Oxcarbazepine was used as an internal standard for this study. The retention times for nevirapine and oxcarbazepine were found to be 7.2 and 14.7 min respectively. The method was found to be linear in the concentration range of 50 ng/mL to 5003.7 ng/mL. The method was validated as per FDA guidelines and was found to be suitable for bioequivalence and pharmacokinetic studies.

  4. Release of leukotriene C4 from human polymorphonuclear leucocytes as determined by radioimmunoassay

    International Nuclear Information System (INIS)

    Aehringhaus, U.; Woelbling, R.H.; Peskar, B.M.; Peskar, B.A.; Koenig, W.; Patrono, C.

    1982-01-01

    Rabbits were immunized with a conjugate of leukotriene (LT)C 4 and bovine serum albumin prepared by coupling the single free amino group of the hapten to the protein using gluteraldehyde. Binding of [ 3 H]LTC 4 to the antibodies obtained is inhibited by 50% with 1.5 ng LTC 4 . The relative cross-section of LTD 4 is 16% and of LTC 4 -methyl ester 3.6%. The validity of the radioimmunoassay was demonstrated by comparison with bioassay using the isolated guinea pig ileum. Using the radioimmunoassay it could be shown that endogenous LTC 4 is released in a dose-dependent manner by human polymorphonuclear leucocytes stimulated with the divalent cation ionophore A23187. (Auth.)

  5. Insights into the ninhydrin chemiluminescent reaction and its potential for micromolar determination of human serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Alvarez, M. Rodriguez [Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, Av. Julian Claveria, 8 33006 Oviedo (Spain); Laino, R. Badia [Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, Av. Julian Claveria, 8 33006 Oviedo (Spain); Diaz-Garcia, M.E. [Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, Av. Julian Claveria, 8 33006 Oviedo (Spain)]. E-mail: medg@uniovi.es

    2006-06-15

    The N-terminal region of human serum albumin (HSA) has an inherent affinity for Co(II) ions. On this basis a new continuous flow method for detection of HSA has been developed taking advantage of the strong quenching effect of the albumin in the ninhydrin-H{sub 2}O{sub 2}-Co(II) chemiluminescent system. The analytical potential of the system is compared with other conventional chemiluminescent reagents. The method gives linear responses from the detection limit (0.30 {mu}M HSA) up to 6.8 {mu}M. The repeatability of the method is good (RSD=7%), it is cheap and rapid to apply and does not require the use of insoluble or expensive reagents nor sophisticated equipment.

  6. Human endogenous retroviruses and multiple sclerosis: innocent bystanders or disease determinants?

    Science.gov (United States)

    Antony, Joseph M; Deslauriers, Andre M; Bhat, Rakesh K; Ellestad, Kristofer K; Power, Christopher

    2011-02-01

    Human endogenous retroviruses (HERVs) constitute 5-8% of human genomic DNA and are replication incompetent despite expression of individual HERV genes from different chromosomal loci depending on the specific tissue. Several HERV genes have been detected as transcripts and proteins in the central nervous system, frequently in the context of neuroinflammation. The HERV-W family has received substantial attention in large part because of associations with diverse syndromes including multiple sclerosis (MS) and several psychiatric disorders. A HERV-W-related retroelement, multiple sclerosis retrovirus (MSRV), has been reported in MS patients to be both a biomarker as well as an effector of aberrant immune responses. HERV-H and HERV-K have also been implicated in MS and other neurological diseases but await delineation of their contributions to disease. The HERV-W envelope-encoded glycosylated protein, syncytin-1, is encoded by chromosome 7q21 and exhibits increased glial expression within MS lesions. Overexpression of syncytin-1 in glia induces endoplasmic reticulum stress leading to neuroinflammation and the induction of free radicals, which damage proximate cells. Syncytin-1's receptor, ASCT1 is a neutral amino acid transporter expressed on glia and is suppressed in white matter of MS patients. Of interest, antioxidants ameliorate syncytin-1's neuropathogenic effects raising the possibility of using these agents as therapeutics for neuroinflammatory diseases. Given the multiple insertion sites of HERV genes as complete and incomplete open reading frames, together with their differing capacity to be expressed and the complexities of individual HERVs as both disease markers and bioactive effectors, HERV biology is a compelling area for understanding neuropathogenic mechanisms and developing new therapeutic strategies. 2010 Elsevier B.V. All rights reserved.

  7. Hydroxychloroquine susceptibility determination of Coxiella burnetii in human embryonic lung (HEL) fibroblast cells.

    Science.gov (United States)

    Angelakis, Emmanouil; Khalil, Jacques Bou; Le Bideau, Marion; Perreal, Celine; La Scola, Bernard; Raoult, Didier

    2017-07-01

    Coxiella burnetii, the causative agent of Q fever, survives and replicates in the acidic environment of monocytes/macrophages; hydroxychloroquine, through alkalinisation of the acidic vacuoles, is critical for the management of Q fever. In this study, a collection of C. burnetii strains isolated from human samples was tested to evaluate the in vitro minimum inhibitory concentrations (MICs) of doxycycline and hydroxychloroquine. Serial two-fold dilutions of doxycycline (0.25-8 mg/L) and hydroxychloroquine (0.25-4 mg/L) were added to C. burnetii-infected human embryonic lung (HEL) fibroblast cells after 48 h of incubation, in duplicate. DNA was detected by C. burnetii-specific semi-quantitative PCR with primers and probes designed for amplification of the IS1111 and IS30A spacers. A total of 29 C. burnetii isolates obtained from 29 patients were tested. Doxycycline MICs ranged from 0.25 mg/L to 0.5 mg/L and hydroxychloroquine MICs from 0.25 mg/L to >4 mg/L. Four C. burnetii stains had hydroxychloroquine MICs ≤ 1 mg/L. The concentration of hydroxychloroquine was associated with a significant decrease in C. burnetii DNA copies in HEL cells based on linear regression analysis (P= 0.01). Recommended serum concentrations of hydroxychloroquine significantly reduced the growth of C. burnetii. Moreover, some C. burnetii strains presented hydroxychloroquine MICs below the recommended serum concentrations, indicating that, for these cases, hydroxychloroquine treatment alone may even be effective. Copyright © 2017 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.

  8. Determination of Radionuclide Concentration in Human Teeth in Najaf Governorate, Iraq

    Directory of Open Access Journals (Sweden)

    Basim Almayahi

    2017-12-01

    Full Text Available Introduction: 238U decays with alpha particles emission into 234Th and the series ends with 206Pb. The unstable nucleus loses the energy with emitting ionizing alpha particles for reaching a stable state. It is undergoing alpha decay with decay energy (4.679 MeV. Alpha particles enter the human and animal bodies through inhalation of air or ingestion of contaminated food and water. This study aimed to perform a radiological analysis on the natural alpha particle emission rates of the human teeth as the biomarkers of radiation exposure and environmental pollution. Materials and Methods: This study was conducted on 68 teeth samples of 27 males and 41 females collected from the hospitals distributed across Najaf governorate, including many districts in Iraq. Alpha particle emission rates were measured using CR-39 nuclear track detector. Results: The mean emission rate of alpha particles in the female teeth was 0.0396±0.0070 mBq cm-2, which was relatively higher than that in the male teeth (0.0390±0.0048 mBq cm-2. Nevertheless, there was no significant difference between the female and male teeth regarding the emission rate of alpha particles. Furthermore, the emission rate of alpha particles in the teeth of the samples taken from Kufa (0.0417±0.0057 mBq cm-2 was higher than those obtained from Najaf (0.0384±0.0053 mBq cm-2. Conclusion: As the findings of this study revealed, Najaf governorate had a lower emission rate of alpha particles as compared to other sites of the worldwide. Therefore, it could be concluded that there is no negative consequence threatening the people’s health in this regard.

  9. 5-fluorouracil Toxicity Mechanism Determination in Human Keratinocytes: in vitro Study on HaCaT

    Directory of Open Access Journals (Sweden)

    Jan Hartinger

    2018-01-01

    Full Text Available 5-fluorouracil (5-FU and capecitabine therapy is often accompanied by palmar-plantar erythrodysesthesia (PPE which is manifestation of 5-FU toxicity in keratinocytes. The main mechanisms of 5-FU action are thymidylate synthase (TS inhibition which can be abrogated by thymidine and strengthened by calciumfolinate (CF and incorporation of fluorouridinetriphosphate into RNA which can be abrogated by uridine. For proper PPE treatment 5-FU mechanism of action in keratinocytes needs to be elucidated. We used the 5-FU toxicity modulators uridine, thymidine and CF to discover the mechanism of 5-FU action in human keratinocyte cell line HaCaT. To measure the cellular viability, we used MTT test and RTCA test. CF did not augment 5-FU toxicity and 5-FU toxicity was weakened by uridine. Therefore, the primary mechanism of 5-FU toxicity in keratinocytes is 5-FU incorporation into RNA. The uridine protective effect cannot fully develop in the presence of CF. Thymidine addition to 5-FU and uridine treated cells not only prevents the toxicity-augmenting CF effect but it also prolongs the 5-FU treated cells survival in comparison to uridine only. Therefore, it can be assumed that in the presence of uridine the 5-FU toxicity mechanism is switched from RNA incorporation to TS inhibition. Although particular 5-FU toxicity mechanisms were previously described in various cell types, this is the first time when various combinations of pyrimidine nucleosides and CF were used for 5-FU toxicity mechanism elucidation in human keratinocytes. We suggest that for PPE treatment ointment containing uridine and thymidine should be further clinically tested.

  10. Micro-bead injection spectroscopy for label-free automated determination of immunoglobulin G in human serum.

    Science.gov (United States)

    Ramos, Inês I; Magalhães, Luís M; Barreiros, Luisa; Reis, Salette; Lima, José L F C; Segundo, Marcela A

    2018-01-01

    Immunoglobulin G (IgG) represents the major fraction of antibodies in healthy adult human serum, and deviations from physiological levels are a generic marker of disease corresponding to different pathologies. Therefore, screening methods for IgG evaluation are a valuable aid to diagnostics. The present work proposes a rapid, automatic, and miniaturized method based on UV-vis micro-bead injection spectroscopy (μ-BIS) for the real-time determination of human serum IgG with label-free detection. Relying on attachment of IgG in rec-protein G immobilized in Sepharose 4B, a bioaffinity column is automatically assembled, where IgG is selectively retained and determined by on-column optical density measurement. A "dilution-and-shoot" approach (50 to 200 times) was implemented without further sample treatment because interferences were flushed out of the column upon sample loading, with minimization of carryover and cross-contamination by automatically discarding the sorbent (0.2 mg) after each determination. No interference from human serum albumin at 60 mg mL -1 in undiluted sample was found. The method allowed IgG determination in the range 100-300 μg mL -1 (corresponding to 5.0-60 mg mL -1 in undiluted samples), with a detection limit of 33 μg mL -1 (1.7 mg mL -1 for samples, dilution factor of 50). RSD values were time-to-result decreased from several hours to times, showing the potential of the proposed approach as a point-of-care method. Graphical abstract Micro-Bead Injection Spectroscopy method for real time, automated and label-free determination of total serum human Immunoglobulin G (IgG). The method was designed for Lab-on-Valve (LOV) platforms using a miniaturised protein G bioaffinity separative approach. IgG are separated from serum matrix components upon quantification with low non-specific binding in less than 5 min.

  11. Non-invasive determination of metabolite concentrations in human transplanted kidney in vivo by 31P MR spectroscopy

    International Nuclear Information System (INIS)

    Kugel, H.; Wittsack, H.J.; Wenzel, F.; Heindel, W.; Lackner, K.; Stippel, D.

    2000-01-01

    To investigate concentrations of phosphorus-containing metabolites in human transplanted kidney in vivo by quantitative 31 P MR spectroscopy (MRS) using surface coils and to compare the obtained values with previous data. Material and Methods: In 5 patients with well-functioning transplanted kidneys, 31 P spectra were obtained with the three-dimensional localization image-selected in vivo spectroscopy technique applying a protocol for quantitative spectroscopy using surface coils. Relaxation corrected signal intensities determined by time domain fitting were used to derive absolute molar concentrations for phosphate-containing metabolites. Results: Little or no phosphocreatine in all spectra verified the absence of muscle contamination, confirming proper volume localization. The mean concentrations in the transplanted kidneys were as follows: ATP 1.60±0.26 mmol/l, PDE 2.14±0.91 mmol/l, Pi 0.66±0.25 mmol/l, PME 2.32±0.50 mmol/l. These values are consistent with previously reported values determined by other techniques. Conclusion: The non-invasive determination of absolute metabolite concentrations in human kidney using MRS supplements the use of signal intensity ratios to detect pathologic changes in the energy metabolism of transplanted kidneys

  12. Energy dispersive x-ray fluorescence spectrometric determination of phosphorus, calcium, iron, zinc, and strontium in human bones

    International Nuclear Information System (INIS)

    Ohta, Akishige; Matsubayashi, Takashi; Itoman, Moritoshi

    1981-01-01

    Phosphorus, calcium, iron, zinc and strontium in a human bone extracted by surgery were determined by energy dispersive X-ray fluorescence spectrometry. The bone was decomposed with nitric acid, then diluted with water. A specific quantity of the solution was naturally dried on polyethylene film, and subjected to X-ray analysis. For determining the calibration curves in a mixture of phosphorus, calcium, iron, zinc and strontium, for the analysis of phosphorus and calcium, germanium was used as the secondary target and aluminum as the filter; and for the analysis of iron, zinc and strontium, molybdenum and molybdenum-aluminum were used, respectively. Consequently, the calibration curves were able to be obtained with high precision in the ranges from 5 to 500 μg of phosphorus, from 1 to 50 μg of calcium and from 0.1 to 1.0 μg of iron, zinc and strontium. In this way, in 1 mg of the human bone by wet weight, phosphorus, calcium, iron, zinc and strontium were able to be determined. (J.P.N.)

  13. Determination of Cd and Pb in human blood by isotope dilution inductively coupled plasma mass spectrometry : International comparison

    International Nuclear Information System (INIS)

    Park, Chang Joon; Suh, Jeong Ki; Lee, Sang Hwa

    1996-01-01

    Inorganic analytical laboratory of Korea Research Institute of Standards and Science participated in an interlaboratory comparison program operated by Quebec Toxicology Centre of Canada in 1994 and again in 1995. The objective of this program is to enable participating laboratories to assess reproducibility and accuracy of their analytical results for trace toxic elements in human biological fluids. This laboratory determined Cd and Pb concentrations in 3 levels of human blood samples by isotope dilution inductively coupled plasma mass spectrometry. 0.5 mL of blood sample is added to the digestion bomb together with 2 mL of nitric acid and enriched spike isotopes and then decomposed in the microwave digestion system. The decomposed sample is diluted to 10 mL and nebulized into ICP-MS. The Cd and Pb values reported by all participating laboratories are presented and compared. The values reported by this laboratory are within the acceptable range of target values. (author)

  14. Ag ion irradiated based sensor for the electrochemical determination of epinephrine and 5-hydroxytryptamine in human biological fluids

    Energy Technology Data Exchange (ETDEWEB)

    Goyal, Rajendra N., E-mail: rngcyfcy@iitr.ernet.in [Department of Chemistry, Indian Institute of Technology Roorkee, Roorkee 247667 (India); Agrawal, Bharati [Department of Chemistry, Indian Institute of Technology Roorkee, Roorkee 247667 (India)

    2012-09-19

    Highlights: Black-Right-Pointing-Pointer Ag ions irradiation enhances the electrocatalytic activity of carbon nano tubes. Black-Right-Pointing-Pointer The low fluence of irradiation caused the ordering of carbon nano tubes. Black-Right-Pointing-Pointer Simultaneous determination of epinephrine and 5-hydroxytryptamine has been carried out. Black-Right-Pointing-Pointer The determination of the neurotransmitters in human blood and urine is reported. - Abstract: A promising and highly sensitive voltammetric method has been developed for the first time for the determination of epinephrine (EP) and 5-hydroxytryptamine (5-HT) using 120 MeV Ag ion irradiated multi-walled carbon nano tube (MWCNT) based sensor. The MWCNT were irradiated at various fluences of 1e12, 3e12 and 1e13 ions cm{sup -2} using palletron accelerator. The simultaneous determination of EP and 5-HT has been carried out in phosphate buffer solution of pH 7.20 using square wave voltammetry and cyclic voltammetry. Experimental results suggested that irradiation of MWCNT by Ag ions enhanced the electrocatalytic activity due to increase in effective surface area and insertion of Ag ions, leading to a remarkable enhancement in peak currents and shift of peak potentials to less positive values as compared to the unirradiated MWCNT (pristine). The developed sensor exhibited a linear relationship between peak current and concentration of EP and 5-HT in the range 0.1-105 {mu}M with detection limit (3{sigma}/b) of 2 nM and 0.75 nM, respectively. The practical utility of irradiation based MWCNT sensor has been demonstrated for the determination of EP and 5-HT in human urine and blood samples.

  15. [Assisted human reproduction techniques: determination of parentage and users in comparative law].

    Science.gov (United States)

    Germán Zurriaráin, Roberto

    2011-01-01

    The paper undertakes the determination of parentage and users of assisted reproduction techniques in comparative law, particularly in three European Union countries: Italy, France and Spain. The idea of artificially conceived child protection, present in the Italian provision, is substituted, partially in French law, and totally in Spanish law, by an exclusively individualistic outlook, configuring a true ″right to maternity″ for married women and for those not living with a male partner (single women, divorced or widowed) or with a female partner (lesbians), and for women inseminated artificially by the semen of the husband before or after their death, and for women already having culminated their fertile life.

  16. Determination of Alpha Activity from Radium-226 and its Decay Products in Human Blood

    International Nuclear Information System (INIS)

    Kenawy, M.A.; Morsy, A.A.; Abdel Hameid, M.M.

    1999-01-01

    During the last few years new and elaborate technique for the quantitative determination of alpha emitting particles in biological samples have been developed using the highly sensitive solid state nuclear track detector CR-39 which allows activities as low as 10 -15 Ci.g -1 to be detected. Blood samples (10 ml) were taken from volunteers. Three pieces of CR-39 were inserted in each blood container and stored at 20 degree for 15 weeks. The pre-etching condition were 6N NaOH+40% C 2 H 5 OH at 60 degree for 3 hours to reduce the background alpha tracks from the environmental radon

  17. Determination of trace elements in human brain tissues using neutron activation analysis

    International Nuclear Information System (INIS)

    Leite, R.E.P.; Jacob-Filho, W.; Grinberg, L.T.; Ferretti, R.E.L.

    2008-01-01

    Neutron activation analysis was applied to assess trace element concentrations in brain tissues from normal (n = 21) and demented individuals (n = 21) of both genders aged more than 50 years. Concentrations of the elements Br, Fe, K, Na, Rb, Se and Zn were determined. Comparisons were made between the results obtained for the hippocampus and frontal cortex tissues, as well as, those obtained in brains of normal and demented individuals. Certified reference materials, NIST 1566b Oyster Tissue and NIST 1577b Bovine Liver were analyzed for quality of the analytical results. (author)

  18. LC-MS/MS method for the determination of clodronate in human plasma.

    Science.gov (United States)

    Hasan, Mahmoud; Schumacher, Gitta; Seekamp, Anne; Taedken, Tobias; Siegmund, Werner; Oswald, Stefan

    2014-11-01

    Clodronate belongs to the class of bisphosphonates which are used for the treatment of bone disorders. Due to its high polarity it has a low and highly variable oral bioavailability which results in low plasma concentrations and requires sensitive bioanalytical methods to characterize its pharmacokinetics in human. Here, we describe for the first time the development and validation of a LC-MS/MS assay for the quantification of clodronate in human plasma. The bisphosphonate was isolated from the biological matrix by protein precipitation using perchloric acid (10%), and derivatized with trimethylorthoacetate prior sample clean-up with liquid-liquid extraction using methyl tert-butyl ether. The chromatography was performed using an isocratic elution with ammonium acetate 5mM (85% v/v, pH 3.8) and acetonitrile (15% v/v) as mobile phase with a flow rate of 300μl/min on a reversed-phase column (Supelco Ascentis(®), C18) temporized at 50°C. The mass spectrometric detection was done using the API4000 triple quadruple mass spectrometer monitoring the mass/charge transitions 301.0/145 for clodronate and 305.2/137.1 for the internal standard etidronate. The analytical range was set to 5-800ng/ml, allowing an evaluation of the plasma concentration-time profiles of clodronate for approximately 7-8 half-life (∼24h). The method was validated according to current FDA/EMA guidelines on bioanalytical method validation with respect to specificity, linearity, intra- and inter-day accuracy and precision, matrix effect, recovery as well as stability. The precision of the assay was 0.6-6.9% and 0.6-8.1% for the intra-day and inter-day variability, respectively. The intra-day and inter-day accuracy (error) was 0.6-8.8% and 2.2-4.5%. The recovery of the analyte was low (2-3%) but reproducible over the entire validation range and sufficient to monitor the target concentrations in human plasma. The drug was shown to be stable in plasma at room temperature for at least 3h (96.0±6%) and

  19. Role of Lysine-54 in determining cofactor specificity and binding in human dihydrofolate reductase

    International Nuclear Information System (INIS)

    Huang, Shaoming; Tan, Xuehai; Thompson, P.D.; Freisheim, J.H.; Appleman, J.R.; Blakley, R.L.; Sheridan, R.P.; Venkataraghavan, R.

    1990-01-01

    Lysine-54 of human dihydrofolate reductase (hDHFR) appears to be involved in the interaction with the 2'-phosphate of NADPH and is conserved as a basic residue in other species. Studies have suggested that in Lactobacillus casei dihydrofolate reductase Arg-43, the homologous residue at this position, plays an important role in the binding of NADPH and in the differentiation of K m values for NADPH and NADH. A Lys-54 to Gln-54 mutant (K54Q) of hDHFR has been constructed by oligodeoxynucleotide-directed mutagenesis in order to study the role of Lys-54 in differentiating K m and k cat values for NADPH and NADH as well as in other functions of hDHFR. The purpose of this paper is to delineate in quantitative terms the magnitude of the effect of the Lys-54 to Gln-54 replacement on the various kinetic parameters of hDHFR. Such quantitative effects cannot be predicted solely on the basis of X-ray structures. The ratio of K m (NADH)/K m (NADPH) decreases from 69 in the wild-type enzyme to 4.7 in the K54Q enzyme, suggesting that Lys-54, among other interactions between protein side-chain residues and the 2'-phosphate, makes a major contribution in terms of binding energy and differentiation of K m values for NADPH and NADH. Agents at concentrations that show activating effects on the wild-type enzyme such as potassium chloride and urea all inactivate the K54Q enzyme. There appear to be no gross conformational differences between wild-type and K54Q enzyme molecules as judged by competitive ELISA using peptide-specific antibodies against human dihydrofolate reductase and from protease susceptibility studies on both wild-type and K54Q mutant enzymes. The pH-rate profiles using NADPH for K54Q and wild-type enzymes show divergences at certain pH values, suggesting the possibility of alteration(s) in the steps of the catalytic pathway for the K54Q enzyme

  20. Commercial radioimmunoassay for beta subunit of human chorionic gonadotropin: falsely positive determinations due to elevated serum luteinizing hormone

    International Nuclear Information System (INIS)

    Fowler, J.E. Jr.; Platoff, G.E.; Kubrock, C.A.; Stuzman, R.E.

    1982-01-01

    Among 17 men who had received seemingly curative treatment for unilateral non-seminomatous germ cell tumors for the testis and who had consistently normal serum human chorionic gonadotropin (HCG) levels at a reference laboratory, 7 (41%) had at least one falsely positive commercial serum HCG determination. To investigate the cause of these falsely positive determinations the authors measured the cross reactivity of luteinizing hormone (LH) and follicle stimulating hormone (FSH) standards in the commercial HCG assay, and studied the relationships between commercial HCG levels and serum LH levels, serum FSH levels and gonadal status in men with and without normal gonadal function. The falsely positive HCG determinations appeared to be due to elevated serum LH levels and cross reactivity of LH in the commercial HCG assay because: 1) there was substantial cross reactivity of the LH standards in the commercial assay, 2) the serum LH was elevated in four of six men with solitary testes, 3) there was a striking correlation between elevated serum LH levels and falsely elevated commercial HCG levels in ten men with solitary or absent testes, and 4) there were no falsely positive HCG determinations in 13 normal men but there were falsely positive HCG determinations in seven of ten anorchid men

  1. [Determination of oxaprozin in human plasma with high performance liquid chromatography (HPLC) and its application].

    Science.gov (United States)

    Mao, Mian; Wang, Ling; Jiang, Xuehua; Yang, Lin

    2013-06-01

    The present research was aimed to develop a high performance liquid chromatography (HPLC) method to determine oxaprozin in plasma and to evaluate the bioavailability of two oxaprozin enteric coated tablets. A C18 column was used to separate the plasma after protein precipitation and the mobile phase was methanol-12. 5mmol/L ammonium acetate buffer solution (pH=3.0)(71:29). The calibration curve was linear in the concentration range of 0. 50-70. 56 microg . mL-1, and the intra and inter-day RSDs were less than 12. 33% and 10. 42% respectively. A single dose of 0. 4 g reference preparation or test preparation of oxaprozin enteric coated tablets was administered to 20 healthy volunteers according to a randomized crossover study. AUC0-->264h were (4 917. 44 +/- 629. 57) microg . h . mL-1 and (4 604. 30+/-737. 83) microg . h . mL-1, respectively; Cmax were (52. 34+/-7. 68) microg . mL-1 and (48. 66+/-4. 87) microg . mL-1, respectively; Tmax were (18. 70+/-2.27) h and (19. 30+/-1. 63) h, respectively; The relative bioavailability of test preparation was 94.0% +/- 13. 7%. The method is simple, rapid and selective for oxaprozin determination. There is no significant difference in the main pharmacokinetic parameters between the test formulation and reference formulation and the two formulations are in bioequivalence.

  2. Determination of fluspirilene in human plasma by liquid chromatography-tandem mass spectrometry with electrospray ionisation.

    Science.gov (United States)

    Swart, K J; Sutherland, F C; van Essen, G H; Hundt, H K; Hundt, A F

    1998-12-18

    An ultra-sensitive method for the determination of fluspirilene in plasma was established, using high-performance liquid chromatographic separation with tandem mass spectrometric detection. The samples were extracted with hexane/isoamyl alcohol, separated on a Phenomenex Luna C18 5 mu 150 x 2.1 mm column with a mobile phase consisting of methanol-water-acetic acid (600:400:1) at a flow-rate of 0.3 ml/min. Detection was achieved by a Finnigan Matt mass spectrometer (LCQ) at unit resolution in full scan mode scanning the product ion spectrum from m/z 130-500 and monitoring the transition of the protonated molecular ion at m/z 476.2, to the sum of the largest product ions m/z 371, 342 and 274 (MS-MS). Electrospray ionisation was used for ion production. The mean recovery for fluspirilene was 90% with a lower limit of quantification of 21.50 pg/ml using 1 ml plasma for extraction. This is the first chromatographic method described for the determination of fluspirilene in plasma that is accurate and sensitive enough to be used in pharmacokinetic studies.

  3. Age and dose as determinants of times of appearance of radium-induced osteosarcomas in humans

    International Nuclear Information System (INIS)

    Stebbings, J.H.; Lucas, H.F.

    1983-01-01

    Determinants of time-until-tumor for bone sarcomas in US radium cases have been reevaluated. For the 57 measured female bone sarcoma cases exposed to 226 Ra and/or 228 Ra, ages at appearance of tumor are more strongly correlated with radium intake doses than are latency periods. Ages at appearance of tumor increase as initial radium intakes decrease. As many lower dose cases were iatrogenic cases exposed in mid-adult life, rather than about age 22 as for the dial worker cases, these results may be artifactual, resulting from an increased susceptability to sarcoma induction at older ages. Alternatively the time of appearance of a sarcoma may be in part directly determined by age. Both explanations find support in the literature. These results suggest that 226 228 Ra-induced bone sarcomas do have an expression period. The downturn in sarcoma incidence at very high doses may result from truncation of the expression period on its early side by the minimum induction period

  4. A Modified LC/MS/MS Method with Enhanced Sensitivity for the Determination of Scopolamine in Human Plasma

    Science.gov (United States)

    Wang, Zuwei; Vaksman, Zalman; Putcha, Lakshmi

    2008-01-01

    Intranasal scopolamine is a choice drug for the treatment of motion sickness during space flight because of its quick onset of action, short half-life and favorable sideeffects profile. The dose administered usually ranges between 0.1 and 0.4 mg. Such small doses make it difficult to detect concentrations of scopolamine in biological fluids using existing sensitive LC/MS/MS method, especially when the biological sample volumes are limited. To measure scopolamine in human plasma to facilitate pharmacokinetic evaluation of the drug, we developed a sensitive LC/MS/MS method using 96 well micro elution plates for solid phase extraction (SPE) of scopolamine in human plasma. Human plasma (100-250 micro L) were loaded onto Waters Oasis HLB 96 well micro elution plate and eluted with 50 L of organic solvent without evaporation and reconstitution. HPLC separation of the eluted sample was performed using an Agilent Zorbax SB-CN column (50 x 2.1 mm) at a flow rate of 0.2 mL/min for 3 minutes. The mobile phase for separation was 80:20 (v/v) methanol: ammonium acetate (30 mM) in water. Concentrations of scopolamine were determined using a Micromass Quattro Micro(TM) mass spectrometer with electrospray ionization (ESI). ESI mass spectra were acquired in positive ion mode with multiple reaction monitoring for the determination of scopolamine m/z = 304.2 right arrow 138.1 and internal standard hyoscyamine m/z = 290.2 right arrow 124.1. The method is rapid, reproducible, specific and has the following parameters: scopolamine and the IS are eluted at about 1.1 and 1.7 min respectively. The linear range is 25-10000 pg/mL for scopolamine in human plasma with correlation coefficients greater than 0.99 and CV less than 0.5%. The intra-day and inter-day CVs are less than 15% for quality control samples with concentrations of 75,300, and 750 pg/mL of scopolamine in human plasma. SPE using 96 well micro elution plates allows rapid sample preparation and enhanced sensitivity for the LC

  5. Which Factors Determine Metal Accumulation in Agricultural Soils in the Severely Human-Coupled Ecosystem?

    Science.gov (United States)

    Xu, Li; Cao, Shanshan; Wang, Jihua; Lu, Anxiang

    2016-05-17

    Agricultural soil is typically an important component of urban ecosystems, contributing directly or indirectly to the general quality of human life. To understand which factors influence metal accumulation in agricultural soils in urban ecosystems is becoming increasingly important. Land use, soil type and urbanization indicators all account for considerable differences in metal accumulation in agricultural soils, and the interactions between these factors on metal concentrations were also examined. Results showed that Zn, Cu, and Cd concentrations varied significantly among different land use types. Concentrations of all metals, except for Cd, were higher in calcareous cinnamon soil than in fluvo-aquic soil. Expansion distance and road density were adopted as urbanization indicators, and distance from the urban center was significantly negatively correlated with concentrations of Hg, and negatively correlated with concentrations of Zn, and road density was positively correlated with Cd concentrations. Multivariate analysis of variance indicated that Hg concentration was significantly influenced by the four-way interaction among all factors. The results in this study provide basic data to support the management of agricultural soils and to help policy makers to plan ahead in Beijing.

  6. Attention Determines Contextual Enhancement versus Suppression in Human Primary Visual Cortex.

    Science.gov (United States)

    Flevaris, Anastasia V; Murray, Scott O

    2015-09-02

    Neural responses in primary visual cortex (V1) depend on stimulus context in seemingly complex ways. For example, responses to an oriented stimulus can be suppressed when it is flanked by iso-oriented versus orthogonally oriented stimuli but can also be enhanced when attention is directed to iso-oriented versus orthogonal flanking stimuli. Thus the exact same contextual stimulus arrangement can have completely opposite effects on neural responses-in some cases leading to orientation-tuned suppression and in other cases leading to orientation-tuned enhancement. Here we show that stimulus-based suppression and enhancement of fMRI responses in humans depends on small changes in the focus of attention and can be explained by a model that combines feature-based attention with response normalization. Neurons in the primary visual cortex (V1) respond to stimuli within a restricted portion of the visual field, termed their "receptive field." However, neuronal responses can also be influenced by stimuli that surround a receptive field, although the nature of these contextual interactions and underlying neural mechanisms are debated. Here we show that the response in V1 to a stimulus in the same context can either be suppressed or enhanced depending on the focus of attention. We are able to explain the results using a simple computational model that combines two well established properties of visual cortical responses: response normalization and feature-based enhancement. Copyright © 2015 the authors 0270-6474/15/3512273-08$15.00/0.

  7. Genetic determination of human facial morphology: links between cleft-lips and normal variation.

    Science.gov (United States)

    Boehringer, Stefan; van der Lijn, Fedde; Liu, Fan; Günther, Manuel; Sinigerova, Stella; Nowak, Stefanie; Ludwig, Kerstin U; Herberz, Ruth; Klein, Stefan; Hofman, Albert; Uitterlinden, Andre G; Niessen, Wiro J; Breteler, Monique M B; van der Lugt, Aad; Würtz, Rolf P; Nöthen, Markus M; Horsthemke, Bernhard; Wieczorek, Dagmar; Mangold, Elisabeth; Kayser, Manfred

    2011-11-01

    Recent genome-wide association studies have identified single nucleotide polymorphisms (SNPs) associated with non-syndromic cleft lip with or without cleft palate (NSCL/P), and other previous studies showed distinctly differing facial distance measurements when comparing unaffected relatives of NSCL/P patients with normal controls. Here, we test the hypothesis that genetic loci involved in NSCL/P also influence normal variation in facial morphology. We tested 11 SNPs from 10 genomic regions previously showing replicated evidence of association with NSCL/P for association with normal variation of nose width and bizygomatic distance in two cohorts from Germany (N=529) and the Netherlands (N=2497). The two most significant associations found were between nose width and SNP rs1258763 near the GREM1 gene in the German cohort (P=6 × 10(-4)), and between bizygomatic distance and SNP rs987525 at 8q24.21 near the CCDC26 gene (P=0.017) in the Dutch sample. A genetic prediction model explained 2% of phenotype variation in nose width in the German and 0.5% of bizygomatic distance variation in the Dutch cohort. Although preliminary, our data provide a first link between genetic loci involved in a pathological facial trait such as NSCL/P and variation of normal facial morphology. Moreover, we present a first approach for understanding the genetic basis of human facial appearance, a highly intriguing trait with implications on clinical practice, clinical genetics, forensic intelligence, social interactions and personal identity.

  8. LC-MS/MS determination of tranexamic acid in human plasma after phospholipid clean-up.

    Science.gov (United States)

    Fabresse, Nicolas; Fall, Fanta; Etting, Isabelle; Devillier, Philippe; Alvarez, Jean-Claude; Grassin-Delyle, Stanislas

    2017-07-15

    Tranexamic acid is a widely used antifibrinolytic drug but its pharmacology and pharmacokinetics remains poorly understood. Owing to the recent knowledge on phospholipid-induced matrix effects during human plasma analysis, our aim was to develop a liquid chromatography-mass spectrometry method for the quantitation of tranexamic acid after efficient sample clean-up. Sample preparation consisted in phospholipid removal and protein precipitation. Hydrophilic interaction liquid chromatography was used and the detection was achieved with multiple reaction monitoring. The method was validated according to the European Medicine Agency guideline in the range 1.0-1000.0μg/mL. The performance of the method was excellent with a precision in the range 1.2-3.0%, an accuracy between 88.4 and 96.6% and a coefficient of variation of the internal standard-normalized matrix factor below 6.7%. This method is suitable for the quantification of tranexamic acid in the wide range of concentrations observed during clinical studies, with all the advantages related to phospholipid removal. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. The 8p23 inversion polymorphism determines local recombination heterogeneity across human populations.

    Science.gov (United States)

    Alves, Joao M; Chikhi, Lounès; Amorim, António; Lopes, Alexandra M

    2014-04-01

    For decades, chromosomal inversions have been regarded as fascinating evolutionary elements as they are expected to suppress recombination between chromosomes with opposite orientations, leading to the accumulation of genetic differences between the two configurations over time. Here, making use of publicly available population genotype data for the largest polymorphic inversion in the human genome (8p23-inv), we assessed whether this inhibitory effect of inversion rearrangements led to significant differences in the recombination landscape of two homologous DNA segments, with opposite orientation. Our analysis revealed that the accumulation of genetic differentiation is positively correlated with the variation in recombination profiles. The observed recombination dissimilarity between inversion types is consistent across all populations analyzed and surpasses the effects of geographic structure, suggesting that both structures (orientations) have been evolving independently over an extended period of time, despite being subjected to the very same demographic history. Aside this mainly independent evolution, we also identified a short segment (350 kb, inversion) in the central region of the inversion where the genetic divergence between the two structural haplotypes is diminished. Although it is difficult to demonstrate it, this could be due to gene flow (possibly via double-crossing over events), which is consistent with the higher recombination rates surrounding this segment. This study demonstrates for the first time that chromosomal inversions influence the recombination landscape at a fine-scale and highlights the role of these rearrangements as drivers of genome evolution.

  10. Secondary structure determination of human. beta. -endorphin by /sup 1/H NMR spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Lichtarge, O.; Jardetzky, O.; Li, C.H.

    1987-09-08

    The /sup 1/H NMR spectra of human ..beta..-endorphin indicate that the peptide exists in random-coil form in aqueous solution but becomes helical in mixed solvent. Thermal denaturation NMR experiments show that in water there is no transition between 24 and 75/sup 0/C, while a slow noncooperative thermal unfolding is observed in a 60% methanol-40% water mixed solvent in the same temperature range. These findings are consistent with circular dichroism studies by other workers concluding that ..beta..-endorphin is a random coil in water but that it forms 50% ..cap alpha..-helix or more in mixed solvents. The peptide in the mixed water-methanol solvent was further studied by correlated spectroscopy (COSY) and nuclear Overhauser effect spectroscopy (NOESY) experiments. These allow a complete set of assignments to be made and establish two distinct stretches over which the solvent induces formation of ..cap alpha..-helices: the first occurs between Tyr-1 and Thr-12 and the second between Leu-14 and extending to Lys-28. There is evidence that the latter is capped by a turn occurring between Lys-28 and Glu-31. These helices form at the enkephalin receptor binding site, which is at the amino terminus, and at the morphine receptor binding site, located at the carboxyl terminus. The findings suggest that these two receptors may specifically recognize ..cap alpha..-helices.

  11. Genetic, anatomic, and clinical determinants of human serum sterol and vitamin D levels.

    Science.gov (United States)

    Stiles, Ashlee R; Kozlitina, Julia; Thompson, Bonne M; McDonald, Jeffrey G; King, Kevin S; Russell, David W

    2014-09-23

    An unknown fraction of the genome participates in the metabolism of sterols and vitamin D, two classes of lipids with diverse physiological and pathophysiological roles. Here, we used mass spectrometry to measure the abundance of >60 sterol and vitamin D derivatives in 3,230 serum samples from a well-phenotyped patient population. Twenty-nine of these lipids were detected in a majority of samples at levels that varied over thousands of fold in different individuals. Pairwise correlations between sterol and vitamin D levels revealed evidence for shared metabolic pathways, additional substrates for known enzymes, and transcriptional regulatory networks. Serum levels of multiple sterols and vitamin D metabolites varied significantly by sex, ethnicity, and age. A genome-wide association study identified 16 loci that were associated with levels of 19 sterols and 25-hydroxylated derivatives of vitamin D (P < 10(-7)). Resequencing, expression analysis, and biochemical experiments focused on one such locus (CYP39A1), revealed multiple loss-of-function alleles with additive effects on serum levels of the oxysterol, 24S-hydroxycholesterol, a substrate of the encoded enzyme. Body mass index, serum lipid levels, and hematocrit were strong phenotypic correlates of interindividual variation in multiple sterols and vitamin D metabolites. We conclude that correlating population-based analytical measurements with genotype and phenotype provides productive insight into human intermediary metabolism.

  12. [Determination of sulpride in human plasma by high performance liquid chromatography].

    Science.gov (United States)

    Yu, X; Luo, Z; Tang, J; Yu, P

    1997-11-01

    This paper describes a reliable method for the pharmacokinetic study of Sulpride in human plasma by reversed-phase high performance liquid chromatography. To compensate the loss of Sulpride during the extraction procedure we used an internal standard very similar in chemical structure and UV absorbance to those of Sulpride. The mobile phase was methanol-water-acetic acid (60:30:1) with a flow rate of 1.2 mL/min. A UV detector was used at 290 nm. The linear range was 5-100 mg/L and the detectable limit was 1.0 mg/L. The recovery and RSD were 97.95%-99.96% and 2.6%-5.1% respectively. The results showed that this method is a sensitive and accurate one which makes the pharmacokinetic study of Sulpride possible. If the concentration was too low to be detected by UV monitor, a fluorescence detector could be used with the excitation wavelength at 299 nm and emission at 342 nm. We analyzed the plasma samples from 30 day-treated psychotic patients and got the satisfactory results.

  13. Neural Determinants of Task Performance during Feature-Based Attention in Human Cortex

    Science.gov (United States)

    Gong, Mengyuan

    2018-01-01

    Abstract Studies of feature-based attention have associated activity in a dorsal frontoparietal network with putative attentional priority signals. Yet, how this neural activity mediates attentional selection and whether it guides behavior are fundamental questions that require investigation. We reasoned that endogenous fluctuations in the quality of attentional priority should influence task performance. Human subjects detected a speed increment while viewing clockwise (CW) or counterclockwise (CCW) motion (baseline task) or while attending to either direction amid distracters (attention task). In an fMRI experiment, direction-specific neural pattern similarity between the baseline task and the attention task revealed a higher level of similarity for correct than incorrect trials in frontoparietal regions. Using transcranial magnetic stimulation (TMS), we disrupted posterior parietal cortex (PPC) and found a selective deficit in the attention task, but not in the baseline task, demonstrating the necessity of this cortical area during feature-based attention. These results reveal that frontoparietal areas maintain attentional priority that facilitates successful behavioral selection. PMID:29497703

  14. Genetics of human body size and shape: pleiotropic and independent genetic determinants of adiposity.

    Science.gov (United States)

    Livshits, G; Yakovenko, K; Ginsburg, E; Kobyliansky, E

    1998-01-01

    The present study utilized pedigree data from three ethnically different populations of Kirghizstan, Turkmenia and Chuvasha. Principal component analysis was performed on a matrix of genetic correlations between 22 measures of adiposity, including skinfolds, circumferences and indices. Findings are summarized as follows: (1) All three genetic matrices were not positive definite and the first four factors retained even after exclusion RG > or = 1.0, explained from 88% to 97% of the total additive genetic variation in the 22 trials studied. This clearly emphasizes the massive involvement of pleiotropic gene effects in the variability of adiposity traits. (2) Despite the quite natural differences in pairwise correlations between the adiposity traits in the three ethnically different samples under study, factor analysis revealed a common basic pattern of covariability for the adiposity traits. In each of the three samples, four genetic factors were retained, namely, the amount of subcutaneous fat, the total body obesity, the pattern of distribution of subcutaneous fat and the central adiposity distribution. (3) Genetic correlations between the retained four factors were virtually non-existent, suggesting that several independent genetic sources may be governing the variation of adiposity traits. (4) Variance decomposition analysis on the obtained genetic factors leaves no doubt regarding the substantial familial and (most probably genetic) effects on variation of each factor in each studied population. The similarity of results in the three different samples indicates that the findings may be deemed valid and reliable descriptions of the genetic variation and covariation pattern of adiposity traits in the human species.

  15. Multiple spatially resolved reflection spectroscopy for in vivo determination of carotenoids in human skin and blood

    Science.gov (United States)

    Darvin, Maxim E.; Magnussen, Björn; Lademann, Juergen; Köcher, Wolfgang

    2016-09-01

    Non-invasive measurement of carotenoid antioxidants in human skin is one of the important tasks to investigate the skin physiology in vivo. Resonance Raman spectroscopy and reflection spectroscopy are the most frequently used non-invasive techniques in dermatology and skin physiology. In the present study, an improved method based on multiple spatially resolved reflection spectroscopy (MSRRS) was introduced. The results obtained were compared with those obtained using the ‘gold standard’ resonance Raman spectroscopy method and showed strong correlations for the total carotenoid concentration (R  =  0.83) as well as for lycopene (R  =  0.80). The measurement stability was confirmed to be better than 10% within the total temperature range from 5 °C to  +  30 °C and pressure contact between the skin and the MSRRS sensor from 800 Pa to 18 000 Pa. In addition, blood samples taken from the subjects were analyzed for carotenoid concentrations. The MSRRS sensor was calibrated on the blood carotenoid concentrations resulting in being able to predict with a correlation of R  =  0.79. On the basis of blood carotenoids it could be demonstrated that the MSRRS cutaneous measurements are not influenced by Fitzpatrick skin types I-VI. The MSRRS sensor is commercially available under the brand name biozoom.

  16. Human capital on the move: Education as a determinant of internal migration in selected INDEPTH surveillance populations in Africa

    Directory of Open Access Journals (Sweden)

    Carren Ginsburg

    2016-05-01

    Full Text Available Background: Education, as a key indicator of human capital, is considered one of the major determinants of internal migration, with previous studies suggesting that human capital accumulates in urban areas at the expense of rural areas. However, there is fragmentary evidence concerning the educational correlates of internal migration in sub-Saharan Africa. Objective: The study questions whether more precise measures of migration in Health and Demographic Surveillance System (HDSS populations support the hypothesis that migrants are self-selected on human capital and more educated people are more likely to leave rural areas or enter urban areas within a geographical region. Methods: Using unique longitudinal data representing approximately 900,000 people living in eight sub-Saharan African HDSS sites that are members of the INDEPTH Network, the paper uses Event History Analysis techniques to examine the relationship between formal educational attainment and in- and out-migration, over the period 2009 to 2011. Results: Between 7Š and 27Š of these local populations are moving in or out of the HDSS area over this period. Education is positively associated with both in- and out-migration in the Kenyan HDSS areas; however, the education effect has no clear pattern in the HDSS sites in Burkina Faso, Mozambique, and South Africa. Conclusions: Empirical results presented in this paper confirm a strong age profile of migration consistent with human capital expectation, yet the results point to variability in the association of education and the propensity to migrate. In particular, the hypothesis of a shift of human capital from rural to urban areas is not universally valid.

  17. Determining tissue-lead levels in large game mammals harvested with lead bullets: human health concerns.

    Science.gov (United States)

    Tsuji, L J S; Wainman, B C; Jayasinghe, R K; VanSpronsen, E P; Liberda, E N

    2009-04-01

    Recently, the use of lead isotope ratios has definitively identified lead ammunition as a source of lead exposure for First Nations people, but the isotope ratios for lead pellets and bullets were indistinguishable. Thus, lead-contaminated meat from game harvested with lead bullets may also be contributing to the lead body burden; however, few studies have determined if lead bullet fragments are present in big game carcasses. We found elevated tissue-lead concentrations (up to 5,726.0 microg/g ww) in liver (5/9) and muscle (6/7) samples of big game harvested with lead bullets and radiographic evidence of lead fragments. Thus, we would advise that the tissue surrounding the wound channel be removed and discarded, as this tissue may be contaminated by lead bullet fragments.

  18. In silico determination of intracellular glycosylation and phosphorylation sites in human selectins: Implications for biological function

    DEFF Research Database (Denmark)

    Ahmad, I.; Hoessli, D.C.; Gupta, Ramneek

    2007-01-01

    Post-translational modifications provide the proteins with the possibility to perform functions in addition to those determined by their primary sequence. However, analysis of multifunctional protein structures in the environment of cells and body fluids is made especially difficult by the presence...... both modifications are likely to occur can also be predicted (YinYang sites), to suggest further functional versatility. Structural modifications of hydroxyl groups of P-, E-, and L-selectins have been predicted and possible functions resulting from such modifications are proposed. Functional changes...... of the three selectins are based on the assumption that transitory and reversible protein modifications by phosphate and O-GlcNAc cause specific conformational changes and generate binding sites for other proteins. The computer-assisted prediction of glycosylation and phosphorylation sites in selectins should...

  19. Use of activation analysis method for determining trace elements in human nail clippings

    International Nuclear Information System (INIS)

    Aguiar, Amilton Reinaldo; Saiki, Mitiko

    1999-01-01

    This work presents results obtained in nail clipping samples analyses. These nail samples were collected from healthy group of individuals and from patients with cystic fibrosis disease. Irradiation of nail clippings was carried out with a thermal neutron flux from 10 11 to 10 13 n.cm -2 .s -1 at the IEA-R1m nuclear research reactor and the elements Al, As, Br, Ca, cd, Cl, Co, Cr, Cu, Fe, K, Mg, Mn, Na, Se and Zn were determined. The results obtained for healthy group were within the range of concentrations published, for normal population. However for several elements, the group of patients presented higher concentrations than those presented for healthy individuals. The precision and the accuracy of the results were evaluated by analyzing reference materials NIST 1577b Bovine Liver and 1566a Oyster Tissue. (author)

  20. Determination of radiation exposure of humans due to dermal contact with radionuclides

    International Nuclear Information System (INIS)

    Pratzel, H.G.

    1992-01-01

    It is the aim of this research project to determine the distribution of radioactive substances in the skin and organism, in order to be able to predict from these data the radiation exposure to be expected following contamination of the skin with radionuclides. Further investigations are to be carried out into the suitability of measures of prevention and decontamination. In view of the fact that only small amounts of those compounds are absorbed by the skin, the determination of the parameters chosen was carried out in radionuclide studies in young pigs. With the exception of the skin of the palms and soles, the radiation effects in man could easily be extrapolated from the results of these experiments. The findings revealed permit the following conclusions to be drawn: The incorporation of radioactive substances after dermal contamination is quite unavoidable, even if this is immediately followed by decontamination measures. The degree of incorporation and resulting radiation dose are only minor and remain several orders of magnitude below the dermal radiation dose. If the skin is contaminated with radioactive liquids, the extent of incorporation and accumulation is proportionate to the activity concentration of those liquids. The incorporated radiation dose is in direct proportion to the contaminated skin area and also shows a relationship to the duration of contamination, if the skin is wet. The uptake by the organism may be somewhat reduced, if decontamination is initiated immediately. Dermal protectives may help to reduce skin permeability and, thus, the uptake of radioactivity into the corneal layer and follicles. (orig./MG) [de

  1. Determination of Drug Toxicity Using 3D Spheroids Constructed From an Immortal Human Hepatocyte Cell Line

    Science.gov (United States)

    Fey, Stephen J.; Wrzesinski, Krzysztof

    2012-01-01

    Numerous publications have documented that the immortal cells grown in three-dimensional (3D) cultures possess physiological behavior, which is more reminiscent of their parental organ than when the same cells are cultivated using classical two-dimensional (2D) culture techniques. The goal of this study was to investigate whether this observation could be extended to the determination of LD50 values and whether 3D data could be correlated to in vivo observations. We developed a noninvasive means to estimate the amount of protein present in a 3D spheroid from it is planar area (± 21%) so that a precise dose can be provided in a manner similar to in vivo studies. This avoided correction of the actual dose given based on a protein determination after treatment (when some cells may have lysed). Conversion of published in vitro LC50 data (mM) for six common drugs (acetaminophen, amiodarone, diclofenac, metformin, phenformin, and valproic acid) to LD50 data (mg compound/mg cellular protein) showed that the variation in LD50 values was generally less than that suggested by the original LC50 data. Toxicological analysis of these six compounds in 3D spheroid culture (either published or presented here) demonstrated similar LD50 values. Although in vitro 2D HepG2 data showed a poor correlation, the primary hepatocyte and 3D spheroid data resulted in a much higher degree of correlation with in vivo lethal blood plasma levels. These results corroborate that 3D hepatocyte cultures are significantly different from 2D cultures and are more representative of the liver in vivo. PMID:22454432

  2. Use of a variable tracer infusion method to determine glucose turnover in humans

    International Nuclear Information System (INIS)

    Molina, J.M.; Baron, A.D.; Edelman, S.V.; Brechtel, G.; Wallace, P.; Olefsky, J.M.

    1990-01-01

    The single-compartment pool fraction model, when used with the hyperinsulinemic glucose clamp technique to measure rates of glucose turnover, sometimes underestimates true rates of glucose appearance (Ra) resulting in negative values for hepatic glucose output (HGO). We focused our attention on isotope discrimination and model error as possible explanations for this underestimation. We found no difference in [3-3H] glucose specific activity in samples obtained simultaneously from the femoral artery and vein (2,400 +/- 455 vs. 2,454 +/- 522 dpm/mg) in 6 men during a hyperinsulinemic euglycemic clamp study where insulin was infused at 40 mU.m-2.min-1 for 3 h; therefore, isotope discrimination did not occur. We compared the ability of a constant (0.6 microCi/min) vs. variable tracer infusion method (tracer added to the glucose infusate) to measure non-steady-state Ra during hyperinsulinemic clamp studies. Plasma specific activity fell during the constant tracer infusion studies but did not change from base line during the variable tracer infusion studies. By maintaining a constant plasma specific activity the variable tracer infusion method eliminates uncertainty about changes in glucose pool size. This overcame modeling error and more accurately measures non-steady-state Ra (P less than 0.001 by analysis of variance vs. constant infusion method). In conclusion, underestimation of Ra determined isotopically during hyperinsulinemic clamp studies is largely due to modeling error that can be overcome by use of the variable tracer infusion method. This method allows more accurate determination of Ra and HGO under non-steady-state conditions

  3. Militarization, human rights violations and community responses as determinants of health in southeastern Myanmar: results of a cluster survey.

    Science.gov (United States)

    Davis, William W; Mullany, Luke C; Schissler, Matt; Albert, Saw; Beyrer, Chris

    2015-01-01

    The Myanmar army and ethnic armed groups agreed to a preliminary ceasefire in 2012, but a heavy military presence remains in southeastern Myanmar. Qualitative data suggested this militarization can result in human rights abuses in the absence of armed engagements between the parties, and that rural ethnic civilians use a variety of self-protection strategies to avoid these abuses or reduce their negative impacts. We used data from a household survey to determine prevalence of select self-protection activities and to examine exposure to armed groups, human rights violations and self-protection activities as determinants of health in southeastern Myanmar. Data collected from 463 households via a two-stage cluster survey of conflict-affected areas in eastern Myanmar in January 2012, were analyzed using logistic regression models to identify associations between exposure to state and non-state armed groups, village self-protection, human rights abuses and health outcomes. Close proximity to a military base was associated with human rights abuses (PRR 1.30, 95 % CI: 1.14-1.48), inadequate food production (PRR 1.08, 95 % CI: 1.03-1.13), inability to access health care (PRR 1.29, 95 % CI: 1.04-1.60) and diarrhea (PRR 1.15, 95 % CI: 1.05-1.27. Direct exposure to armed groups was associated with household hunger (PRR1.71, 95 % CI: 1.30-2.23). Among households that reported no human rights abuses, risk of household hunger (PRR 5.64, 95 % CI: 1.88-16.91), inadequate food production (PRR 1.95, 95 % CI: 1.11-3.41) and diarrhea (PRR 2.53, 95 % CI: 1.45-4.42) increased when neighbors' households reported experiencing human rights abuses. Households in villages that reported negotiating with the Myanmar army had lower risk of human rights violations (PRR 0.91, 95 % CI: 0.85-0.98), household hunger (PRR 0.85, 95 % CI: 0.74-0.96), inadequate food production (PRR 0.93, 95 % CI:0.89-0.98) and diarrhea (PRR 0.89, 95 % CI:0.82-0.97). Stratified analysis suggests that self

  4. Socio-economic and cultural determinants of human african trypanosomiasis at the Kenya - Uganda transboundary.

    Directory of Open Access Journals (Sweden)

    Jane Jemeli Rutto

    Full Text Available BACKGROUND: Kenya and Uganda have reported different Human African Trypanosomiasis incidences in the past more than three decades, with the latter recording more cases. This cross-sectional study assessed the demographic characteristics, tsetse and trypanosomiasis control practices, socio-economic and cultural risk factors influencing Trypanosoma brucei rhodesiense (T.b.r. infection in Teso and Busia Districts, Western Kenya and Tororo and Busia Districts, Southeast Uganda. A conceptual framework was postulated to explain interactions of various socio-economic, cultural and tsetse control factors that predispose individuals and populations to HAT. METHODS: A cross-sectional household survey was conducted between April and October 2008. Four administrative districts reporting T.b.r and lying adjacent to each other at the international boundary of Kenya and Uganda were purposely selected. Household data collection was carried out in two villages that had experienced HAT and one other village that had no reported HAT case from 1977 to 2008 in each district. A structured questionnaire was administered to 384 randomly selected household heads or their representatives in each country. The percent of respondents giving a specific answer was reported. Secondary data was also obtained on socio-economic and political issues in both countries. RESULTS: Inadequate knowledge on the disease cycle and intervention measures contributed considerable barriers to HAT, and more so in Uganda than in Kenya. Gender-associated socio-cultural practices greatly predisposed individuals to HAT. Pesticides-based crop husbandry in the 1970's reportedly reduced vector population while vegetation of coffee and banana's and livestock husbandry directly increased occurrence of HAT. Livestock husbandry practices in the villages were strong predictors of HAT incidence. The residents in Kenya (6.7% applied chemoprophylaxis and chemotherapeutic controls against trypanosomiasis to a

  5. The eIF4AIII RNA helicase is a critical determinant of human cytomegalovirus replication

    International Nuclear Information System (INIS)

    Ziehr, Ben; Lenarcic, Erik; Cecil, Chad; Moorman, Nathaniel J.

    2016-01-01

    Human cytomegalovirus (HCMV) was recently shown to encode a large number of spliced mRNAs. While the nuclear export of unspliced viral transcripts has been extensively studied, the role of host mRNA export factors in HCMV mRNA trafficking remains poorly defined. We found that the eIF4AIII RNA helicase, a component of the exon junction complex, was necessary for efficient virus replication. Depletion of eIF4AIII limited viral DNA accumulation, export of viral mRNAs from the nucleus, and the production of progeny virus. However eIF4AIII was dispensable for the association of viral transcripts with ribosomes. We found that pateamine A, a natural compound that inhibits both eIF4AI/II and eIF4AIII, has potent antiviral activity and inhibits HCMV replication throughout the virus lytic cycle. Our results demonstrate that eIF4AIII is required for efficient HCMV replication, and suggest that eIF4A family helicases may be a new class of targets for the development of host-directed antiviral therapeutics. - Highlights: • The host eIF4AIII RNA helicase is required for efficient HCMV replication. • Depleting eIF4AIII inhibited the nuclear export of HCMV mRNAs. • HCMV mRNAs did not require eIF4AIII to associate with polyribosomes. • The eIF4A family helicases may be new targets for host-directed antiviral drugs.

  6. The eIF4AIII RNA helicase is a critical determinant of human cytomegalovirus replication

    Energy Technology Data Exchange (ETDEWEB)

    Ziehr, Ben; Lenarcic, Erik; Cecil, Chad; Moorman, Nathaniel J., E-mail: nmoorman@med.unc.edu

    2016-02-15

    Human cytomegalovirus (HCMV) was recently shown to encode a large number of spliced mRNAs. While the nuclear export of unspliced viral transcripts has been extensively studied, the role of host mRNA export factors in HCMV mRNA trafficking remains poorly defined. We found that the eIF4AIII RNA helicase, a component of the exon junction complex, was necessary for efficient virus replication. Depletion of eIF4AIII limited viral DNA accumulation, export of viral mRNAs from the nucleus, and the production of progeny virus. However eIF4AIII was dispensable for the association of viral transcripts with ribosomes. We found that pateamine A, a natural compound that inhibits both eIF4AI/II and eIF4AIII, has potent antiviral activity and inhibits HCMV replication throughout the virus lytic cycle. Our results demonstrate that eIF4AIII is required for efficient HCMV replication, and suggest that eIF4A family helicases may be a new class of targets for the development of host-directed antiviral therapeutics. - Highlights: • The host eIF4AIII RNA helicase is required for efficient HCMV replication. • Depleting eIF4AIII inhibited the nuclear export of HCMV mRNAs. • HCMV mRNAs did not require eIF4AIII to associate with polyribosomes. • The eIF4A family helicases may be new targets for host-directed antiviral drugs.

  7. What determines the acceptability of genetically modified food that can improve human nutrition?

    Science.gov (United States)

    Purchase, Iain F H

    2005-09-01

    It has been predicted that by 2025 there will be an annual shortfall of cereals for feeding the human population of 68.5 million tones. One possible solution is the use of genetically modified (GM) crops, which are already grown extensively (59 million ha of GM crops were planted in 2002) in the USA, South America, Africa and China. Nevertheless, there is considerable disagreement about the advisability of using such crops, particularly in Europe. Obviously, the safety of the food derived from the GM crops is a primary consideration. Safety assessment relies on establishing that the food is substantially equivalent to its non-GM counterpart and specific testing for allergenicity of proteins and toxicity of metabolites and the whole food. There appears to be international agreement on the principles of safety assessment. Safety to the environment is equally important, but will not be covered in this presentation. The public's perception of the risk of new technology is critical to its acceptance. Perception of risk, in turn, depends on the credibility of the source of the information and trust in the regulatory process. In many countries, the public appears to have lost its trust in the scientists and government dealing with GM food, making the acceptability of GM crops uncertain. Of equal importance are the socio-economic factors that impinge on the viability of GM produce. These include intellectual property protection, trade liberalization (through subsidy and tariff barriers in developed countries) and the intensity of bio safety regulations. The socio-economic interests of developed and developing countries may diverge and may even be contradictory in any one country. Acceptance of GM crops will thus depend on detailed issues surrounding particular crops and economies.

  8. What determines the acceptability of genetically modified food that can improve human nutrition?

    International Nuclear Information System (INIS)

    Purchase, Iain F.H.

    2005-01-01

    It has been predicted that by 2025 there will be an annual shortfall of cereals for feeding the human population of 68.5 million tonnes. One possible solution is the use of genetically modified (GM) crops, which are already grown extensively (59 million ha of GM crops were planted in 2002) in the USA, South America, Africa and China. Nevertheless, there is considerable disagreement about the advisability of using such crops, particularly in Europe. Obviously, the safety of the food derived from the GM crops is a primary consideration. Safety assessment relies on establishing that the food is substantially equivalent to its non-GM counterpart and specific testing for allergenicity of proteins and toxicity of metabolites and the whole food. There appears to be international agreement on the principles of safety assessment. Safety to the environment is equally important, but will not be covered in this presentation. The public's perception of the risk of new technology is critical to its acceptance. Perception of risk, in turn, depends on the credibility of the source of the information and trust in the regulatory process. In many countries, the public appears to have lost its trust in the scientists and government dealing with GM food, making the acceptability of GM crops uncertain. Of equal importance are the socio-economic factors that impinge on the viability of GM produce. These include intellectual property protection, trade liberalisation (through subsidy and tariff barriers in developed countries) and the intensity of bio safety regulations. The socio-economic interests of developed and developing countries may diverge and may even be contradictory in any one country. Acceptance of GM crops will thus depend on detailed issues surrounding particular crops and economies

  9. What determines the acceptability of genetically modified food that can improve human nutrition?

    Energy Technology Data Exchange (ETDEWEB)

    Purchase, Iain F.H. [University of Manchester, Oxford Road, Manchester M13 9PT (United Kingdom)

    2005-09-01

    It has been predicted that by 2025 there will be an annual shortfall of cereals for feeding the human population of 68.5 million tonnes. One possible solution is the use of genetically modified (GM) crops, which are already grown extensively (59 million ha of GM crops were planted in 2002) in the USA, South America, Africa and China. Nevertheless, there is considerable disagreement about the advisability of using such crops, particularly in Europe. Obviously, the safety of the food derived from the GM crops is a primary consideration. Safety assessment relies on establishing that the food is substantially equivalent to its non-GM counterpart and specific testing for allergenicity of proteins and toxicity of metabolites and the whole food. There appears to be international agreement on the principles of safety assessment. Safety to the environment is equally important, but will not be covered in this presentation. The public's perception of the risk of new technology is critical to its acceptance. Perception of risk, in turn, depends on the credibility of the source of the information and trust in the regulatory process. In many countries, the public appears to have lost its trust in the scientists and government dealing with GM food, making the acceptability of GM crops uncertain. Of equal importance are the socio-economic factors that impinge on the viability of GM produce. These include intellectual property protection, trade liberalisation (through subsidy and tariff barriers in developed countries) and the intensity of bio safety regulations. The socio-economic interests of developed and developing countries may diverge and may even be contradictory in any one country. Acceptance of GM crops will thus depend on detailed issues surrounding particular crops and economies.

  10. Determination of material emission signatures by PTR-MS and their correlations with odor assessments by human subjects

    DEFF Research Database (Denmark)

    K H, Han; J S, Zhang; Wargocki, Pawel

    2010-01-01

    by human subjects. VOC emissions from each material were measured in a 50-l small-scale chamber. Chamber air was sampled by PTR-MS to determine emission signatures. Sorbent tube sampling and TD-GC/MS analysis were also performed to identify the major VOCs emitted and to compare the resulting data...... VOC odor indices was used to represent the emission level measured by PTR-MS.......The objectives of this study were to determine volatile organic compound (VOC) emission signatures of nine typical building materials by using proton transfer reaction-mass spectrometry (PTR-MS) and to explore the correlation between the PTR-MS measurements and the measurements of acceptability...

  11. Silver nanoparticles plasmon resonance-based method for the determination of uric acid in human plasma and urine samples

    International Nuclear Information System (INIS)

    Amjadi, M.; Rahimpour, E.

    2012-01-01

    We have developed a simple and sensitive colorimetric procedure for the quantification of trace amounts of uric acid. It is based on the finding that uric acid in a medium containing ammonia and sodium hydroxide at 65 0 C can reduce silver ions to form yellow silver nanoparticles (Ag NPs). These are stabilized in solution by using poly(vinyl alcohol) as a capping agent. The yellow color of the solution that results from the localized surface plasmon resonance of Ag NPs can be observed by the bare eye. The absorbance at 415 nm is proportional to the concentration of uric acid which therefore can be determined quantitatively. The calibration curve is linear in the concentration range from 10 to 200 nM, with a limit of detection of 3.3 nM. The method was successfully applied to the determination of uric acid in human plasma and urine samples. (author)

  12. Trace element content of human hair determined using neutron activation analysis as monitor of exposure effects to environmental metals

    International Nuclear Information System (INIS)

    Dutkiewicz, T.; Paprotny, W.; Sokolowska, D.; Kulka, E.; Woyciechowska, E.; Dybczynski, R.; Sterlinski, S.

    1978-01-01

    Determination of 17 trace elements, mainly metals, in small hair samples by neutron activation analysis was elaborated. The number of elements determined in one sample made it possible to assess quickly the exposure of human population in a given region. The hair samples after careful washing according to procedure elaborated previously were irradiated in high reactor neutron flux (approximately 10 14 n.cm -2 .s -1 ). The activities of samples were measured three times (for different decay times) by means of Ge(Li) gamma-ray spectrometer. A new version of the comparator technique, called a triple-single comparator method, was applied for quantitative analysis. The statistical methods used for planning of sampling and preliminary results on evaluation of analytical data are also described. (author)

  13. How do social determinants affect human trafficking in Southeast Asia, and what can we do about it? A systematic review.

    Science.gov (United States)

    Perry, Kelsey McGregor; McEwing, Lindsay

    2013-12-12

    The sale of women and children accounts for the greatest proportion of human trafficking globally, with Southeast Asia acting as the illegal industry's largest international hub. At least 225,000 women and children are trafficked from the region every year, accounting for approximately one-third of the global human trade. The health ramifications of trafficking are severe: many survivors contract infectious diseases including sexually transmitted infections and develop mental health conditions, including anxiety, panic disorder, and major depression. The complications associated with studying a highly secretive illegal trade have severely limited research on effective prevention measures. Because this presents a challenge for organizations that hope to develop prevention strategies, we asked the following question: How do social determinants facilitate or mitigate trafficking of women and children in Southeast Asia, and what recommendations does the literature provide for combating trafficking via these social determinants? Using a Cochrane-based systematic search methodology, five independent researchers reviewed 1,148 articles from the past ten years (2001–2011). After three phases of independent review, they selected and analyzed 61 articles to identify the determinants that impact trafficking of women and children in Southeast Asia. Key social determinants that facilitate trafficking include poverty, female gender, lack of policy and enforcement, age, migration, displacement and conflict, ethnicity, culture, ignorance of trafficking methods, and caste status. Conversely, protective determinants that mitigate trafficking include formal education, citizenship, maternal education, higher caste status, and birth order. Recommendations relating to a variety of the determinants are identified and discussed in detail. Social determinants are central to the processes that mitigate and facilitate the sale and exploitation of women and children in Southeast Asia

  14. [Diagnostic significance of serum free DNA human telomerase reverse transcriptase quantitative determination on spinal cord injury].

    Science.gov (United States)

    Yang, M K; Tang, J; Xiang, Z; Zhang, X; Wang, J; Li, Z; Li, Y; Sheng, W B

    2018-02-06

    Objective: To investigate the relationship between the content of human telomerase reverse transcriptase (hTERT) and its clinical features in serum free DNA in patients with different degree of spinal cord injury. Methods: From December 2013 to December 2016, inpatients of the Central Hospital of Bazhong, Sichuan Province were enrolledand divided into the experimental group, the disease control group and the negative control group. For the experimental group: 46 patients with spinal cord injury were graded according to the criteria of the American Association of Spinal Cord Injury (ASIA), including 12 cases of grade A, 10 cases of grade B, 10 cases of grade C, 7 cases of grade D and 7 cases of grade E; for the disease control group: 15 patients with spinal fractures (without spinal cord injury) at the same period were included; and for the negative control group: 20 healthy adult volunteers aged 18-50 years were selected.Real-time fluorescence quantitative PCR and immunoblotting were performed to detect the content of hTERT in serum free DNA both in patients and healthy controls and to compare the difference between them. The results of the somatosensory evoked potential (SEP) of all patients were compared and analyzed.The receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of hTERT content in serum free DNA in patients with spinal cord injury. Results: Comparison of serum free DNA hTERT content: in the experimental group, the serum free DNA hTERT content of grade A, B, C, D, E was (99.63±8.23), (76.24±4.37), (46.07±5.43), (16.30±0.95) and (15.74±1.12)μg/L, respectively.While it was (15.01±1.39)μg/L in the disease control group and (14.54±1.03)μg/L in the negative control group. The total difference was statistically significant between patients of each group and the control group ( F =857.917, P spinal cord injury has a certain guiding significance for the diagnosis of spinal cord injury and the degree of injury.

  15. Determination of hydroxyurea in human plasma by HPLC-UV using derivatization with xanthydrol.

    Science.gov (United States)

    Legrand, Tiphaine; Rakotoson, Marie-Georgine; Galactéros, Frédéric; Bartolucci, Pablo; Hulin, Anne

    2017-10-01

    A simple and rapid high performance liquid chromatography (HPLC) method using ultraviolet (UV) detection was developed to determine hydroxyurea (HU) concentration in plasma sample after derivatization with xanthydrol. Two hundred microliters samples were spiked with methylurea (MeU) as internal standard and proteins were precipitated by adding methanol. Derivatization of HU and MeU was immediately performed by adding 0.02M xanthydrol and 1.5M HCl in order to obtain xanthyl-derivatives of HU and MeU that can be further separated using HPLC and quantified using UV detection at 240nm. Separation was achieved using a C18 column with a mobile phase composed of 20mM ammonium acetate and acetonitrile in gradient elution mode at a flow rate of 1mL/min. The total analysis time did not exceed 18min. The method was found linear from 5 to 400μM and all validation parameters fulfilled the international requirements. Between- and within-run accuracy error ranged from -4.7% to 3.2% and precision was lower than 12.8%. This simple method requires small volume samples and can be easily implemented in most clinical laboratories to develop pharmacokinetics studies of HU and to promote its therapeutic monitoring. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Bioanalytical methods for the determination of cocaine and metabolites in human biological samples.

    Science.gov (United States)

    Barroso, M; Gallardo, E; Queiroz, J A

    2009-08-01

    Determination of cocaine and its metabolites in biological specimens is of great importance, not only in clinical and forensic toxicology, but also in workplace drug testing. These compounds are normally screened for using sensitive immunological methods. However, screening methods are unspecific and, therefore, the posterior confirmation of presumably positive samples by a specific technique is mandatory. Although GC-MS-based techniques are still the most commonly used for confirmation purposes of cocaine and its metabolites in biological specimens, the advent of LC-MS and LC-MS/MS has enabled the detection of even lower amounts of these drugs, which assumes particular importance when sample volume available is small, as frequently occurs with oral fluid. This paper will review recently-published papers that describe procedures for detection of cocaine and metabolites, not only in the most commonly used specimens, such as blood and urine, but also in other 'alternative' matrices (e.g., oral fluid and hair) with a special focus on sample preparation and chromatographic analysis.

  17. Atomic Resolution Structures of Human Bufaviruses Determined by Cryo-Electron Microscopy

    Directory of Open Access Journals (Sweden)

    Maria Ilyas

    2018-01-01

    Full Text Available Bufavirus strain 1 (BuV1, a member of the Protoparvovirus genus of the Parvoviridae, was first isolated from fecal samples of children with acute diarrhea in Burkina Faso. Since this initial discovery, BuVs have been isolated in several countries, including Finland, the Netherlands, and Bhutan, in pediatric patients exhibiting similar symptoms. Towards their characterization, the structures of virus-like particles of BuV1, BuV2, and BuV3, the current known genotypes, have been determined by cryo-electron microscopy and image reconstruction to 2.84, 3.79, and 3.25 Å, respectively. The BuVs, 65–73% identical in amino acid sequence, conserve the major viral protein, VP2, structure and general capsid surface features of parvoviruses. These include a core β-barrel (βB-βI, α-helix A, and large surface loops inserted between these elements in VP2. The capsid contains depressions at the icosahedral 2-fold and around the 5-fold axes, and has three separated protrusions surrounding the 3-fold axes. Structure comparison among the BuVs and to available parvovirus structures revealed capsid surface variations and capsid 3-fold protrusions that depart from the single pinwheel arrangement of the animal protoparvoviruses. These structures provide a platform to begin the molecular characterization of these potentially pathogenic viruses.

  18. Assessment of the usefulness of the PIGE method to determine nitrogen in human blood serum

    International Nuclear Information System (INIS)

    Kupila-Rantala, T.; Hyvoenen-Dabek, M.; Raiasanen, J.; Dabek, J.T.

    1995-01-01

    Proton-induced gamma-ray emission (PIGE) analysis has been applied to the determination of total nitrogen concentration of blood sera from hospital patients. Both the reaction 14 N(p,p'γ) 14 N, E γ =2313 keV, and the reaction 15 N(p,αγ) 12 C,E γ =4439keV, have been used. From 37 patients, the range of the nitrogen concentrations obtained was 9.54-16.3 g/l with a mean of 12.0 g/l. The corresponding total protein concentrations varied between 59 and 96 g/l, measured in a clinical laboratory auto analyser using the biuret method. The range of the albumin concentrations was 29-46 g/l. The nitrogen concentrations correlated well with the total protein concentrations (r=0.802). The multiple correlation of nitrogen versus (albumin+ (total protein- albumin)) gives a coefficient of 0.175 for albumin and 0.161 for total protein- albumin. The results agree with expectations and support the potential usefulness of the rapid PIGE method for medical studies. (author). 17 refs., 3 figs

  19. Spectrofluorimetric determination of aliskiren in tablets and spiked human plasma through derivatization with dansyl chloride.

    Science.gov (United States)

    Aydoğmuş, Zeynep; Sarı, Ferhat; Ulu, Sevgi Tatar

    2012-03-01

    A simple and sensitive method has been developed and validated for the determination of aliskiren (ALS) in its dosage forms and spiked plasma. The method was based on the reaction of the drug with dansyl chloride in the presence of bicarbonate solution of pH 10.5 to give a highly fluorescent derivative which was measured at 501 nm with excitition at 378 nm in dichloromethane. Different experimental parameters affecting the development of the method and stability were carefully studied and optimized. The calibration curves were linear over the concentration ranges of 100-700 and 50-150 ng/mL for standard solution and plasma, respectively. The limits of detection were 27.52 ng/mL in standard solution, 4.91 ng/mL in plasma. The developed method was successfully applied to the analysis the drug in the commercial tablets and spiked plasma samples. The mean recovery of ALS from tablets and plasma was 100.10 and 97.81%, respectively. A proposal of the reaction pathway was presented.

  20. [Determination of ciprofloxacin in human serum and urine by reversed-phase HPLC].

    Science.gov (United States)

    Qin, Y; Liang, D

    1993-03-01

    A sensitive and rapid method for the determination of ciprofloxacin using enoxacin as the internal standard was reported. High-performance liquid chromatograph model 344 (Beckman, USA) with a variable wavelength UV detector and reversed-phase Ultrasphere-ODS column (5 microns, 250 x 4.6 mm) was used. Serum or urine sample preparation involved addition phosphate buffer (pH 7.5) and aqueous solution of sodium lauryl sulfate, followed by chloroform extraction. The organic layer was removed and evaporated to dryness under an air stream in a 37 degrees C water bath. The residue was dissolved in 50 microliters mobile phase and 20 microliters injected. The mobile phase of 0.02 mol/L acetate buffer (pH 3.0) -acetonitril-dimethylformamide-10% aqueous solution of tetrabutyl ammonium hydroxide (88:6.5:5:0.5) was pumped at 0.9 ml/min through the column. The detector operated at 0.01 aufs and the wavelength was set at 276 nm. The retention times for ciprofloxacin and enoxacin were 7.31 min and 5.59 min, respectively. In serum, standard curve was linear in the concentration range of 0.75 to 24 mumol/L, the detective limit was 0.2 mumol/L, extraction recovery was 69-74%, within-day CV was less than 5%, and inter-day CV was less than 6%.

  1. Determination of total mercury and methylmercury in human head hair by radiochemical methods of analysis

    International Nuclear Information System (INIS)

    Vasconcellos, M.B.A.; Saiki, M.; Paletti, G.; Pinheiro, R.M.M.; Baruzzi, R.G.; Spindel, R.

    1992-01-01

    Efforts were carried out in order to detect population groups in Brazil that could be at risk with respect to mercury contamination, mainly by ingestion of contaminated fish. Two regions were identified. The first was near Billings Dam which is located in one of the most heavily industrialized parts of the country. It is suspected that the water is polluted by chlor-alkali and other industrial pollutants. People living near this dam frequently consume fish caught around or near the dam. The second region is Xingu Park, located in the Amazonic region where several Indian tribes live and where the gold exploration activities have caused much concern due to the use of mercury in the ore processing. Tons of mercury are thrown in the rivers of the region annually. Both of these groups and a control group were used in this study. It was concluded that the population near Billings Dam had normal levels of mercury. However, the Indians from Xingu Park had very high levels of mercury in their Hair. It was determined that this group deserved further study. 7 refs, 2 tabs

  2. Trace elements determinations in cancerous and non-cancerous human tissues using instrumental neutron activation analysis

    International Nuclear Information System (INIS)

    Choi, Insup.

    1989-01-01

    Recent improvements in analyzing techniques when coupled to the growing knowledge of trace element biochemistry provide a powerful tool to investigate the relationship between trace elements and cancer. It is hoped that selective delivery or restriction of specific minerals may aid in cancer prevention or treatment. Tissues were collected at the time of surgery of various cancer patients including colon cancer and breast cancer. Three kinds of tissues were taken from a patient; cancerous, noncancerous, and transitional tissue obtained from a region located between the cancer and healthy tissues. A total of 57 tissues were obtained from 19 cancer patients. Seven of them were colon cancer patients, and 5 of them were breast cancer patients. Nine elements were determined using instrumental activation analysis. Cancerous colon tissue had significantly higher concentrations of selenium and iron than healthy tissues. Cancerous breast tissue had significantly higher concentrations of selenium, iron, manganese, and rubidium than healthy tissues. Iron can be enriched in cancer tissue because cancer tissue retains more blood vessels. Selenium is enriched in cancer tissue, possibly in an effort of the body to inhibit the growth of tumors. The manganese enrichment can be explained in the same manner as selenium considering its suspected anticarcinogenicity. It is not certain why rubidium was enriched in cancer tissue. It could be that this is the result of alteration of cell membrane permeability, change in extracellular matrix, or increased metabolism in cancer tissue

  3. Opportunities and limits in applying epidemiologic methods to determine the effects of ionizing radiation in humans

    International Nuclear Information System (INIS)

    Lilienfeld, A.M.

    1980-01-01

    In this presentation, an attempt has been made to indicate rather broadly some of the opportunities and limits of the epidemiologic studies of ionizing radiation effects. The review has not been exhaustive. However, it is clear that much further information is needed before estimates of radiation effects can be made with the kind of confidence that one desires. These scientific considerations should be divorced from the policy considerations where a more conservative attitude would appear desirable. Epidemiologic studies now in progress should be continued on the survivors in Hiroshima and Nagasaki. An attempt should be made to locate other exposed groups in order to obtain information to determine the types and frequencies of malignancies and other possible effects. In epidemiologic studies of the effect of radiation, there should be incorporated mechanisms for obtaining information on other possible carcinogenic factors to detemine the possible effects of interaction between radiation and these other factors. Conversely, in studies of other types of carcinogenic agents, data on radiation exposure should also be obtained. To increase the value of such studies, it is important to develop a standardized method of obtaining information on radiation exposure as well as some means of categorizing such exposure

  4. Use of DNA from bite marks can determine species and individual animals that attack humans

    Science.gov (United States)

    Farley, Sean; Talbot, Sandra L.; Sage, George K.; Sinnott, Rick; Coltrane, Jessica

    2014-01-01

    During the summer of 2008, 6 documented attacks and close encounters with brown bears (Ursus arctos) occurred in the greater Anchorage, Alaska (USA) area. We discuss findings from 2 incidents in which people were mauled within 2 km of each other over a 6-week period and in which it was assumed that a single animal was responsible. To ensure public safety, authorities killed a brown bear implicated in the attacks by circumstantial evidence, though it was not known a priori that the animal was responsible. We extracted DNA from hairs and bite sites on the clothing of both victims and determined species and individual identity of the animal(s) involved in both incidents. Genetic data indicated the brown bear killed by authorities was responsible for one of the maulings, but not both. This research demonstrates that DNA-based techniques, with appropriate sampling, can provide unambiguous identification of animals involved in attacks, as well as provide reasonable justification for excluding others. Because DNA-based techniques can unequivocally identify individual bears carrying out attacks, they should be considered a standard method employed in wildlife attack investigations.

  5. Ultrasensitive determination of mercury in human saliva by atomic fluorescence spectrometry based on solidified floating organic drop microextraction

    International Nuclear Information System (INIS)

    Yuan, C.-G.; Wang, J.; Jin, Y.

    2012-01-01

    We report on a new, rapid and simple method for the determination of ultra-trace quantities of mercury ion in human saliva. It is based on solidified floating organic drop microextraction and detection by cold vapor atomic fluorescence spectrometry (CV-AFS). Mercury ion was complexed with diethyldithiocarbamate, and the hydrophobic complex was then extracted into fine droplets of 1-undecanol. By cooling in an ice bath after extraction, the droplets in solution solidify to form a single ball floating on the surface of solution. The solidified micro drop containing the mercury complex was then transferred for determination by CV-AFS. The effects of pH value, concentration of chelating reagent, quantity of 1-undecanol, sample volume, equilibration temperature and time were investigated. Under the optimum conditions, the preconcentration of a 25-mL sample is accomplished with an enrichment factor of 182. The limit of detection is 2.5 ng L -1 . The relative standard deviation for seven replicate determinations at 0.1 ng mL -1 level is 4.1%. The method was applied to the determination of mercury in saliva samples collected from four volunteers. Two volunteers having dental amalgam fillings had 0.4 ng mL -1 mercury in their saliva, whereas mercury was not detectable in the saliva of two volunteers who had no dental fillings. (author)

  6. Improvements on enzymatic hydrolysis of human hair for illicit drug determination by gas chromatography/mass spectrometry.

    Science.gov (United States)

    Míguez-Framil, Martha; Moreda-Piñeiro, Antonio; Bermejo-Barrera, Pilar; López, Patricia; Tabernero, María Jesús; Bermejo, Ana María

    2007-11-15

    The use of ultrasound energy for accelerating the pronase E enzymatic hydrolysis of human hair for extracting illicit drugs has been novelty tested. The enzymatic extracts obtained after 30 min of sonication in an ultrasonic water bath were subjected to an optimized solid-phase extraction process, which involved a solution of 2.0% (v/v) acetic acid in methanol as eluting solution and concentration by N2 stream evaporation. A gas chromatography/mass spectrometry method was used to separate and determine cocaine, benzoylecgonine, codeine, morphine, and 6-monoacethylmorphine in 20 min. Variables affecting ultrasound-assisted pronase E hydrolysis such as hydrolysis temperature, hydrolysis time, enzyme concentration, catalyzer (1,4-dithiothreitol) concentration, ionic strength, pH, and ultrasound frequency were simultaneously evaluated by a Plackett-Burman design 2(8) PBD of resolution III. The most statistically significant variables were ionic strength and pH, which means that analyte extraction is mainly attributed to pronase E activity. The optimization or evaluation of all the factors has led to an accelerated pronase E hydrolysis of human hair, which can be completed in 30 min. Results have been found to be statistically similar to those obtained with conventional pronase E hydrolysis. The accelerated method was finally applied to several human hair samples from multidrug abusers.

  7. Dual-Energy Computed Tomography Gemstone Spectral Imaging: A Novel Technique to Determine Human Cardiac Calculus Composition.

    Science.gov (United States)

    Cheng, Ching-Li; Chang, Hsiao-Huang; Ko, Shih-Chi; Huang, Pei-Jung; Lin, Shan-Yang

    2016-01-01

    Understanding the chemical composition of any calculus in different human organs is essential for choosing the best treatment strategy for patients. The purpose of this study was to assess the capability of determining the chemical composition of a human cardiac calculus using gemstone spectral imaging (GSI) mode on a single-source dual-energy computed tomography (DECT) in vitro. The cardiac calculus was directly scanned on the Discovery CT750 HD FREEdom Edition using GSI mode, in vitro. A portable fiber-optic Raman spectroscopy was also applied to verify the quantitative accuracy of the DECT measurements. The results of spectral DECT measurements indicate that effective Z values in 3 designated positions located in this calculus were 15.02 to 15.47, which are close to values of 15.74 to 15.86, corresponding to the effective Z values of calcium apatite and hydroxyapatite. The Raman spectral data were also reflected by the predominant Raman peak at 960 cm for hydroxyapatite and the minor peak at 875 cm for calcium apatite. A potential single-source DECT with GSI mode was first used to examine the morphological characteristics and chemical compositions of a giant human cardiac calculus, in vitro. The CT results were consistent with the Raman spectral data, suggesting that spectral CT imaging techniques could be accurately used to diagnose and characterize the compositional materials in the cardiac calculus.

  8. Capillary electrophoresis with indirect UV detection for the determination of stabilizers and citrates present in human albumin solutions.

    Science.gov (United States)

    Jaworska, Małgorzata; Cygan, Paulina; Wilk, Małgorzata; Anuszewska, Elzbieta

    2009-08-15

    Sodium caprylate and N-acetyltryptophan are the most frequently used stabilizers that protect the albumin from aggregation or heat induced denaturation. In turn citrates - excipients remaining after fractionation process - can be treated as by-product favoring leaching aluminum out of glass containers whilst albumin solution is stored. With ionic nature these substances have all the markings of a subject for capillary electrophoresis analysis. Thus CE methods were proposed as new approach for quality control of human albumin solution in terms of determination of stabilizers and citrates residue. Human albumin solutions both 5% and 20% from various manufacturers were tested. Indirect detection mode was set to provide sufficient detectability of analytes lacking of chromophores. As being anions analytes were separated with reversed electroosmotic flow. As a result of method optimization two background electrolytes based on p-hydroxybenzoic acid and 2,6-pyridinedicarboxylic acid were selected for stabilizers and citrates separation, respectively. The optimized methods were successfully validated. For citrates that require quantification below 100microM the method demonstrated the precision less than 4% and the limit of detection at 4microM. In order to check the new methods accuracy and applicability the samples were additionally tested with selected reference methods. The proposed methods allow reliable quantification of stabilizers and citrates in human albumin solution that was confirmed by method validation as well as result comparison with reference methods. The CE methods are considered to be suitable for quality control yet simplifying and reducing cost of analysis.

  9. Evaluation of capillary zone electrophoresis for the determination of protein composition in therapeutic immunoglobulins and human albumins.

    Science.gov (United States)

    Christians, Stefan; van Treel, Nadine Denise; Bieniara, Gabriele; Eulig-Wien, Annika; Hanschmann, Kay-Martin; Giess, Siegfried

    2016-07-01

    Capillary zone electrophoresis (CZE) provides an alternative means of separating native proteins on the basis of their inherent electrophoretic mobilities. The major advantage of CZE is the quantification by UV detection, circumventing the drawbacks of staining and densitometry in the case of gel electrophoresis methods. The data of this validation study showed that CZE is a reliable assay for the determination of protein composition in therapeutic preparations of human albumin and human polyclonal immunoglobulins. Data obtained by CZE are in line with "historical" data obtained by the compendial method, provided that peak integration is performed without time correction. The focus here was to establish a rapid and reliable test to substitute the current gel based zone electrophoresis techniques for the control of protein composition of human immunoglobulins or albumins in the European Pharmacopoeia. We believe that the more advanced and modern CZE method described here is a very good alternative to the procedures currently described in the relevant monographs. Copyright © 2016 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.

  10. A Human Rights-Based Approach to Farmworker Health: An Overarching Framework to Address the Social Determinants of Health.

    Science.gov (United States)

    Ramos, Athena K

    2018-01-01

    Migrant and seasonal workers have a right to the highest attainable standard of health. Unfortunately, these farmworkers face a multitude of challenges. They are employed in one of the most dangerous industries and face serious occupational health risks, while positioned at the bottom of the social hierarchy. They often lack formal education and training, English language proficiency, legal status, access to information, and equitable opportunities to health and healthcare. This article will explore the international human rights conventions that support farmworkers' right to health and healthcare in the United States. International human rights may provide a valuable legal framework that could be used to advocate on behalf of farmworkers and address the social determinants of health. Therefore, a Human Rights-Based Approach to Farmworker health will be presented along with recommendations for how to advance health and access to healthcare among this population. Fostering the health and well-being of migrant and seasonal farmworkers is critical to advancing equity, social justice, and maintaining the workforce required to meet production needs and safeguard the economic competitiveness of the industry.

  11. Picomolar concentrations of morphine in human urine determined by dansyl derivatization and liquid chromatography-mass spectrometry.

    Science.gov (United States)

    Lamshöft, Marc; Grobe, Nadja; Spiteller, Michael

    2011-04-15

    Morphine is present in varying amounts as an endogenous product in human urine. Derivatization of morphine contained in urine with dansyl chloride yields a known product, which can be quantified by liquid chromatography mass spectrometry with high selectivity and sensitivity. Urine samples of 51 healthy individuals were spiked with stable-isotope labeled morphine, hydrolyzed and subjected to solid phase extraction followed by derivatization of morphine with dansyl chloride. The dansyl derivatives of naturally occurring morphine and deuterated internal standard were then detected by liquid chromatography-triple quadrupole mass spectrometry. Using the [N-CD(3)]-labeled internal standard and solid-phase extraction, a limit of detection of 35 fmol/ml (10 pg/ml) and a limit of quantification of 87.5 fmol/ml (25 pg/ml) was determined for morphine in human urine. This new LC-MS/MS method allowed the detection of endogenous morphine in human urine of 51 volunteers with an average value of 156.4 fmol/ml (44.7 ng/ml). Copyright © 2011 Elsevier B.V. All rights reserved.

  12. Determination of the optical properties of melanin-pigmented human skin equivalents using terahertz time-domain spectroscopy

    Science.gov (United States)

    Lipscomb, Dawn; Echchgadda, Ibtissam; Peralta, Xomalin G.; Wilmink, Gerald J.

    2013-02-01

    Terahertz time-domain spectroscopy (THz-TDS) methods have been utilized in previous studies in order to characterize the optical properties of skin and its primary constituents (i.e., water, collagen, and keratin). However, similar experiments have not yet been performed to investigate whether melanocytes and the melanin pigment that they synthesize contribute to skin's optical properties. In this study, we used THz-TDS methods operating in transmission geometry to measure the optical properties of in vitro human skin equivalents with or without normal human melanocytes. Skin equivalents were cultured for three weeks to promote gradual melanogenesis, and THz time domain data were collected at various time intervals. Frequency-domain analysis techniques were performed to determine the index of refraction (n) and absorption coefficient (μa) for each skin sample over the frequency range of 0.1-2.0 THz. We found that for all samples as frequency increased, n decreased exponentially and the μa increased linearly. Additionally, we observed that skin samples with higher levels of melanin exhibited greater n and μa values than the non-pigmented samples. Our results indicate that melanocytes and the degree of melanin pigmentation contribute in an appreciable manner to the skin's optical properties. Future studies will be performed to examine whether these contributions are observed in human skin in vivo.

  13. The determination of polycyclic aromatic hydrocarbons in human urine by high-resolution gas chromatography-mass spectrometry.

    Science.gov (United States)

    Cho, Sung-Hee; Lee, Sun-Kyung; Kim, Chong Hyeak

    2018-05-01

    Polycyclic aromatic hydrocarbons (PAHs), organic compounds formed by at least two condensed aromatic rings, are ubiquitous environmental pollutants that are produced by incomplete combustion of organic materials. PAHs have been classified as carcinogenIC to humans by the International Agency for Research on Cancer, because they can bind to DNA, causing mutations. Therefore, the levels of PAHs in human urine can be used as an indicator for potential carcinogenesis and cell mutation. An analytical method was developed for the accurate measurement of PAHs in urine using high-resolution gas chromatography-mass spectrometry. Urine samples were extracted by an Oasis HLB extraction cartridge after enzymatic hydrolysis with a β-glucuronidase/arylsulfatase cocktail. The 18 PAHs were separated using an Agilent DB-5 MS capillary column (30 m × 0.25 mm, 0.25 μm) and monitored by time-of-flight mass spectrometry. Under the optimized method, the linearity of calibration curves was >0.994. The limits of detection at a signal-to-noise ratio of 3 were 10-100 ng/L. The coefficients of variation were in the range of 0.4-9.0%. The present method was highly accurate for simultaneous determination of 18 PAHs in human urine and could be applied to monitoring and biomedical investigations to check exposure of PAHs. Copyright © 2017 John Wiley & Sons, Ltd.

  14. Radiosensitivity of different human tumor cells lines grown as multicellular spheroids determined from growth curves and survival data

    International Nuclear Information System (INIS)

    Schwachoefer, J.H.C.; Crooijmans, R.P.; van Gasteren, J.J.; Hoogenhout, J.; Jerusalem, C.R.; Kal, H.B.; Theeuwes, A.G.

    1989-01-01

    Five human tumor cell lines were grown as multicellular tumor spheroids (MTS) to determine whether multicellular tumor spheroids derived from different types of tumors would show tumor-type dependent differences in response to single-dose irradiation, and whether these differences paralleled clinical behavior. Multicellular tumor spheroids of two neuroblastoma, one lung adenocarcinoma, one melanoma, and a squamous cell carcinoma of the oral tongue, were studied in terms of growth delay, calculated cell survival, and spheroid control dose50 (SCD50). Growth delay and cell survival analysis for the tumor cell lines showed sensitivities that correlated well with clinical behavior of the tumor types of origin. Similar to other studies on melanoma multicellular tumor spheroids our spheroid control dose50 results for the melanoma cell line deviated from the general pattern of sensitivity. This might be due to the location of surviving cells, which prohibits proliferation of surviving cells and hence growth of melanoma multicellular tumor spheroids. This study demonstrates that radiosensitivity of human tumor cell lines can be evaluated in terms of growth delay, calculated cell survival, and spheroid control dose50 when grown as multicellular tumor spheroids. The sensitivity established from these evaluations parallels clinical behavior, thus offering a unique tool for the in vitro analysis of human tumor radiosensitivity

  15. Determination of the 4-monohydroxy metabolites of perhexiline in human plasma, urine and liver microsomes by liquid chromatography.

    Science.gov (United States)

    Davies, Benjamin J; Herbert, Megan K; Coller, Janet K; Somogyi, Andrew A; Milne, Robert W; Sallustio, Benedetta C

    2006-11-07

    The use of perhexiline (PHX) is limited by hepatic and neurological toxicity associated with elevated concentrations in plasma that are the result of polymorphism of the cytochrome P450 2D6 isoform (CYP2D6). PHX is cleared by hepatic oxidation that produces three 4-monohydroxy metabolites: cis-OH-PHX, trans1-OH-PHX and trans2-OH-PHX. The current study describes an HPLC-fluorescent method utilising pre-column derivatization with dansyl chloride. Following derivatization, the metabolites were resolved on a C18 column with a gradient elution using a mobile phase composed of methanol and water. The method described is suitable for the quantification of the metabolites in human plasma and urine following clinical doses and for kinetic studies using human liver microsomes. The method demonstrates sufficient sensitivity, accuracy and precision between 5.0 and 0.01, 50.0 and 0.2 and 1.0 and 0.005 mg/l in human plasma, urine and liver microsomes, respectively, with intra-assay coefficients of variation and bias D6 extensive metaboliser (EM) patients at steady state with respect to PHX dosing determined that the mean (+/-S.D.) renal clearances of trans1-OH-PHX and cis-OH-PHX were 1.58+/-0.35 and 0.16+/-0.06l/h, respectively. The mean (+/-S.D.) dose recovered in urine as free and glucuronidated 4-monohydroxy PHX metabolites was 20.6+/-11.6%.

  16. A novel liquid chromatography/mass spectrometry method for determination of neurotransmitters in brain tissue: Application to human tauopathies.

    Science.gov (United States)

    Forgacsova, Andrea; Galba, Jaroslav; Garruto, Ralph M; Majerova, Petra; Katina, Stanislav; Kovac, Andrej

    2018-01-15

    Neurotransmitters, small molecules widely distributed in the central nervous system are essential in transmitting electrical signals across neurons via chemical communication. Dysregulation of these chemical signaling molecules is linked to numerous neurological diseases including tauopathies. In this study, a precise and reliable liquid chromatography method was established with tandem mass spectrometry detection for the simultaneous determination of aspartic acid, asparagine, glutamic acid, glutamine, γ-aminobutyric acid, N-acetyl-l-aspartic acid, pyroglutamic acid, acetylcholine and choline in human brain tissue. The method was successfully applied to the analysis of human brain tissues from three different tauopathies; corticobasal degeneration, progressive supranuclear palsy and parkinsonism-dementia complex of Guam. Neurotransmitters were analyzed on ultra-high performance chromatography (UHPLC) using an ethylene bridged hybrid amide column coupled with tandem mass spectrometry (MS/MS). Identification and quantification of neurotransmitters was carried out by ESI+ mass spectrometry detection. We optimized sample preparation to achieve simple and fast extraction of all nine analytes. Our method exhibited an excellent linearity for all analytes (all coefficients of determination >0.99), with inter-day and intra-day precision yielding relative standard deviations 3.2%-11.2% and an accuracy was in range of 92.6%-104.3%. The present study, using the above method, is the first to demonstrate significant alterations of brain neurotransmitters caused by pathological processes in the brain tissues of patient with three different tauopathies. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Comparison of two extraction procedures for determination of drugs of abuse in human saliva by high-performance liquid chromatography.

    Science.gov (United States)

    Fernández, P; Morales, L; Vázquez, C; Lago, M; Bermejo, A M

    2008-11-01

    High performance liquid chromatography in combination with diode array detection (HPLC-DAD) was used to determine morphine, 6-acetylmorphine, cocaine, benzoylecgonine, cocaethylene, methadone and 2-ethylene-1,5-dimethyl-3,3,-diphenylpyrrolidine in