scholarly journals Lymphocyte metallothionein mRNA responds to marginal zinc intake in human volunteers

2000 ◽  
Vol 84 (5) ◽  
pp. 747-756 ◽  
Author(s):  
Adrian K. Allan ◽  
Gabrielle M. Hawksworth ◽  
Leslie R. Woodhouse ◽  
Barbara Sutherland ◽  
Janet C. King ◽  
...  
Keyword(s):  
Human Subjects ◽  
Baseline Period ◽  
Zn Deficiency ◽  
Pcr Assay ◽  
Laser Induced Fluorescence Detection ◽  
Pcr Product ◽  
Human Volunteers ◽  
Actin Mrna ◽  
Zn Status ◽  
Diagnostic Indicator

Marginal Zn deficiency is thought to be prevalent in both developed and developing countries. However, the extent of Zn deficiency is not known, due to the lack of a reliable diagnostic indicator. Blood plasma and erythrocyte concentrations of metallothionein (MT) reflect Zn status, but measurement of MT is dependent on the availability of sensitive immunoassays. Our aim was to show whether measurement of T lymphocyte MT-2A mRNA, using a competitive reverse transcriptase (RT)–polymerase chain reaction (PCR) assay, could indicate Zn status in human subjects in a residential Zn-depletion study. In the study, the Zn intake of seven volunteers was maintained at 13·7 mg/d for 5 weeks (baseline) followed by 4·6 mg/d for 10 weeks (marginal intake) and then 13·7 mg/d (repletion) for 5 weeks. The quantitative assay was developed using standard techniques and concentrations of MT-2A mRNA were normalized by reference to β-actin mRNA which was also measured by competitive RT–PCR assay. An alternative method of measuring the PCR product using capillary electrophoresis with laser-induced fluorescence detection was also evaluated. There was considerable inter-individual variation in MT-2A mRNA concentration and the mean level at the end of the baseline period was 10·3 (SE 3·7) fg MT-2A mRNA/pg β-actin mRNA, which then decreased by 64 % during the low Zn intake period. After repletion, MT-2A mRNA returned to baseline concentrations. In contrast, plasma Zn was unchanged by marginal Zn intake or repletion. The effect of low Zn in all individuals was consistent. We conclude that this assay is a sensitive method of evaluating marginal changes in dietary Zn intake.

scholarly journals BIMG-13. A NOVEL RADIOPHARMACEUTICAL ([18F]DASA-23) TO MONITOR PYRUVATE KINASE M2 INDUCED GLYCOLYTIC REPROGRAMMING IN GLIOBLASTOMA

2020 ◽  
Vol 3 (Supplement_1) ◽  
pp. i3-i4
Author(s):  
Corinne Beinat ◽  
Chirag Patel ◽  
Tom Haywood ◽  
Surya Murty ◽  
Lewis Naya ◽  
...  
Keyword(s):  
Cell Culture ◽  
Pyruvate Kinase ◽  
Cancer Metabolism ◽  
Human Subjects ◽  
Normal Brain ◽  
Healthy Human ◽  
Pyruvate Kinase M2 ◽  
Molar Activity ◽  
Pet Tracer ◽  
Human Volunteers

Abstract BACKGROUND Pyruvate kinase M2 (PKM2) catalyzes the final step in glycolysis, a key process of cancer metabolism. PKM2 is preferentially expressed by glioblastoma (GBM) cells with minimal expression in healthy brain, making it an important biomarker of cancer glycolytic re-programming. We describe the bench-to-bedside development, validation, and translation of a novel positron emission tomography (PET) tracer to study PKM2 in GBM. Specifically, we evaluated 1-((2-fluoro-6-[18F]fluorophenyl)sulfonyl)-4-((4-methoxyphenyl)sulfonyl)piperazine ([18F]DASA-23) in cell culture, mouse models of GBM, healthy human volunteers, and GBM patients. METHODS [18F]DASA-23 was synthesized with a molar activity of 100.47 ± 29.58 GBq/µmol and radiochemical purity >95%. We performed initial testing of [18F]DASA-23 in GBM cell culture and human GBM xenografts implanted orthotopically into mice. Next we produced [18F]DASA-23 under current Good Manufacturing Practices United States Food and Drug Administration (FDA) oversight, and evaluated it in healthy volunteers and a pilot cohort of patients with gliomas. RESULTS In mouse imaging studies, [18F]DASA-23 clearly delineated the U87 GBM from the surrounding healthy brain tissue and had a tumor-to-brain ratio (TBR) of 3.6 ± 0.5. In human volunteers, [18F]DASA-23 crossed the intact blood-brain barrier and was rapidly cleared. In GBM patients, [18F]DASA-23 successfully outlined tumors visible on contrast-enhanced magnetic resonance imaging (MRI). The uptake of [18F]DASA-23 was markedly elevated in GBMs compared to normal brain, and it was able to identify a metabolic non-responder within 1-week of treatment initiation. CONCLUSION We developed and translated [18F]DASA-23 as a promising new tracer that demonstrated the visualization of aberrantly expressed PKM2 for the first time in human subjects. These encouraging results warrant further clinical evaluation of [18F]DASA-23 to assess its utility for imaging therapy-induced normalization of aberrant cancer metabolism.

Sensitivity and reliability of zinc transporter and metallothionein gene expression in peripheral blood mononuclear cells as indicators of zinc status: responses to ex vivo zinc exposure and habitual zinc intake in humans

2020 ◽  
pp. 1-8
Author(s):  
Stephen R. Hennigar ◽  
Alyssa M. Kelley ◽  
Bradley J. Anderson ◽  
Nicholes J. Armstrong ◽  
Holly L. McClung ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Ex Vivo ◽  
Human Subjects ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
Essential Nutrient ◽  
Zn Status

Abstract Zn is an essential nutrient for humans; however, a sensitive biomarker to assess Zn status has not been identified. The objective of this study was to determine the reliability and sensitivity of Zn transporter and metallothionein (MT) genes in peripheral blood mononuclear cells (PBMCs) to Zn exposure ex vivo and to habitual Zn intake in human subjects. In study 1, human PBMCs were cultured for 24 h with 0–50 µm ZnSO4 with or without 5 µm N,N,N′,N′-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN), and mRNA expression of SLC30A1-10, SLC39A1-14, MT1 subtypes (A, B, E, F, G, H, L, M and X), MT2A, MT3 and MT4 mRNA was determined. In study 2, fifty-four healthy male and female volunteers (31·9 (sd 13·8) years, BMI 25·7 (sd 2·9) kg/m2) completed a FFQ, blood was collected, PBMCs were isolated and mRNA expression of selected Zn transporters and MT isoforms was determined. Study 1: MT1E, MT1F, MT1G, MT1H, MT1L, MT1M, MT1X, MT2A and SLC30A1 increased with increasing concentrations of Zn and declined with the addition of TPEN. Study 2: Average daily Zn intake was 16·0 (sd 5·3) mg/d (range: 9–31 mg/d), and plasma Zn concentrations were 15·5 (SD 2·8) μmol/l (range 11–23 μmol/l). PBMC MT2A was positively correlated with dietary Zn intake (r 0·306, P = 0·03) and total Zn intake (r 0·382, P < 0·01), whereas plasma Zn was not (P > 0·05 for both). Findings suggest that MT2A mRNA in PBMCs reflects dietary Zn intake in healthy adults and may be a component in determining Zn status.

scholarly journals A schlieren optical study of the human cough with and without wearing masks for aerosol infection control

2009 ◽  
Vol 6 (suppl_6) ◽  
Author(s):  
Julian W. Tang ◽  
Thomas J. Liebner ◽  
Brent A. Craven ◽  
Gary S. Settles
Keyword(s):  
High Speed ◽  
Fluid Dynamic ◽  
Human Subjects ◽  
Tracer Gas ◽  
Airborne Infection ◽  
Dynamic Mechanisms ◽  
Human Volunteers ◽  
Natural Mechanism ◽  
Time Changes ◽  
Aerosol Infection

Various infectious agents are known to be transmitted naturally via respiratory aerosols produced by infected patients. Such aerosols may be produced during normal activities by breathing, talking, coughing and sneezing. The schlieren optical method, previously applied mostly in engineering and physics, can be effectively used here to visualize airflows around human subjects in such indoor situations, non-intrusively and without the need for either tracer gas or airborne particles. It accomplishes this by rendering visible the optical phase gradients owing to real-time changes in air temperature. In this study, schlieren video records are obtained of human volunteers coughing with and without wearing standard surgical and N95 masks. The object is to characterize the exhaled airflows and evaluate the effect of these commonly used masks on the fluid-dynamic mechanisms that spread infection by coughing. Further, a high-speed schlieren video of a single cough is analysed by a computerized method of tracking individual turbulent eddies, demonstrating the non-intrusive velocimetry of the expelled airflow. Results show that human coughing projects a rapid turbulent jet into the surrounding air, but that wearing a surgical or N95 mask thwarts this natural mechanism of transmitting airborne infection, either by blocking the formation of the jet (N95 mask), or by redirecting it in a less harmful direction (surgical mask).

A gold nanoparticles-assisted multiplex PCR assay for simultaneous detection of Salmonella typhimurium, Listeria monocytogenes and Escherichia coli O157:H7

2020 ◽  
Vol 12 (2) ◽  
pp. 212-217 ◽  
Author(s):  
Juan Du ◽  
Shujing Wu ◽  
Liyuan Niu ◽  
Junguang Li ◽  
Dianbo Zhao ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gold Nanoparticles ◽  
Listeria Monocytogenes ◽  
Salmonella Typhimurium ◽  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Escherichia Coli O157 ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Pcr Product

Unfunctionalized flower-shaped AuNPs is used as colorimetric sensor for PCR product detection by naked eyes.

scholarly journals Detection of Trypanosoma cruzi and Trypanosoma rangeli Infection by Duplex PCR Assay Based on Telomeric Sequences

2003 ◽  
Vol 10 (5) ◽  
pp. 775-779 ◽  
Author(s):  
Miguel Angel Chiurillo ◽  
Gladys Crisante ◽  
Agustina Rojas ◽  
Andreina Peralta ◽  
Manuel Dias ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Human Subjects ◽  
Simultaneous Detection ◽  
Duplex Pcr ◽  
Trypanosoma Rangeli ◽  
Pcr Assay ◽  
Telomeric Sequences ◽  
Species Specific ◽  
Duplex Pcr Assay ◽  
Reduviid Bugs

ABSTRACT We used the species specificity and repetitious nature of subtelomeric kinetoplastida sequences to generate a duplex PCR assay for the simultaneous detection of Trypanosoma cruzi and Trypanosoma rangeli in experimentally and naturally infected triatomine (Reduviid) bugs and in infected human subjects. The assay was species specific and was capable of detecting 1/20th of T. cruzi and 1/4th of T. rangeli cell equivalents without complementary hybridization. In addition, the PCR-based assay was robust enough for direct application to difficult biological samples such as Reduviid feces or guts and was capable of recognizing all T. cruzi and T. rangeli strains and lineages. Because the assay primers amplify entirely different target sequences, no reaction interference was observed, facilitating future adaptation of this assay to an automated format.

Polymerase Chain Reaction Assay for Detection of Escherichia coli O157:H7 and Escherichia coli O157:H−

2002 ◽  
Vol 65 (1) ◽  
pp. 5-11 ◽  
Author(s):  
TAKAHISA MIYAMOTO ◽  
NATSUKO ICHIOKA ◽  
CHIE SASAKI ◽  
HIROSHI KOBAYASHI ◽  
KEN-ICHI HONJOH ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Dna Sequence ◽  
Escherichia Coli O157 ◽  
Chain Reaction ◽  
Pcr Assay ◽  
E Coli ◽  
Pcr Product ◽  
Genomic Dnas ◽  
Polymerase Chain

The DNA band patterns generated by polymerase chain reaction (PCR) using the du2 primer and template DNAs from various strains of Escherichia coli and non–E. coli bacteria were compared. Among three to five prominent bands produced, the three bands at about 1.8, 2.7, and 5.0 kb were detected in all of the E. coli O157 strains tested. Some nonpathogenic E. coli and all pathogenic E. coli except E. coli O157 showed bands at 1.8 and 5.0 kb. It seems that the band at 2.7 kb is specific to E. coli O157. Sequence analysis of the 2.7-kb PCR product revealed the presence of a DNA sequence specific to E. coli O157:H− and E. coli O157:H7. Since the DNA sequence from base 15 to base 1008 of the PCR product seems to be specific to E. coli O157, a PCR assay was carried out with various bacterial genomic DNAs and O157-FHC1 and O157-FHC2 primers that amplified the region between base 23 and base 994 of the 2.7-kb PCR product. A single band at 970 bp was clearly detected in all of the strains of E. coli O157:H− and E. coli O157:H7 tested. However, no band was amplified from template DNAs from other bacteria, including both nonpathogenic and pathogenic E. coli except E. coli O157. All raw meats inoculated with E. coli O157:H7 at 3 × 100 to 3.5 × 102 CFU/25 g were positive both for our PCR assay after cultivation in mEC-N broth at 42°C for 18 h and for the conventional cultural method.

scholarly journals The role of ZIP proteins in zinc assimilation and distribution in plants: current challenges

2019 ◽  
Vol 55 (No. 2) ◽  
pp. 45-54 ◽  
Author(s):  
Juan Daniel Lira-Morales ◽  
Nancy Varela-Bojórquez ◽  
Magaly Berenice Montoya-Rojo ◽  
J. Adriana Sañudo-Barajas
Keyword(s):  
Family Members ◽  
Zn Deficiency ◽  
Mineral Concentration ◽  
Regulation Of Expression ◽  
Zip Family ◽  
Zn Homeostasis ◽  
Homeostasis Regulation ◽  
Zn Status ◽  
Nutritional Imbalance

Soils with mineral deficiencies lead to nutritional imbalance in crops worldwide. Zinc (Zn) is a micronutrient that is fundamental for plant growth and development, being essential for the proper functioning of a range of enzymes and transcription factors. Zn transporters tightly regulate Zn homeostasis. Plants contain a large number of Zn-responsive genes that are specifically expressed under Zn deficiency to ensure the coordination of assimilatory pathways and meet the physiological requirements. This review brings together a range of studies that have been undertaken to investigate the effects of Zn status on the regulatory mechanisms involved in plant mineral nutrition. The ZIP (ZRT, IRT-like Protein) family is especially implicated in Zn transport and in the maintenance of cellular Zn homeostasis. Regulation of expression in relation to plant tissue, mineral concentration, and species has been determined for several ZIP family members. In the omic era, genomic and proteomic approaches have facilitated a rapid increase in our understanding of the roles of ZIP family members and their regulation, though significant knowledge gaps remain. A comprehensive understanding of ZIP proteins could lead to many potential molecular applications to improve crop management and food quality.  

scholarly journals Erythrocytes, erythrocyte membranes, neutrophils and platelets as biopsy materials for the assessment of zinc status in humans

1992 ◽  
Vol 68 (2) ◽  
pp. 515-527 ◽  
Author(s):  
Manuel Ruz ◽  
Kelley R. Cavan ◽  
William J. Bettger ◽  
Rosalind S. Gibson
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Erythrocyte Membranes ◽  
Zn Deficiency ◽  
Molar Ratios ◽  
Potential Index ◽  
Zinc Depletion ◽  
Zn Status ◽  
Cellular Immune

During a controlled zinc depletion-repletion study, fifteen men aged 25.3 (sd 3.3) years were fed on a low-Zn diet with high phytate:Zn and phytate × calcium: Zn molar ratios for 7 weeks, followed by a 2 week repletion period when 30 mg supplemental Zn/d was given. Changes in plasma, urine, and hair Zn concentrations, taste acuity, and cellular immune response confirmed the development of mild Zn deficiency. Zn concentrations in neutrophils, platelets, erythrocytes and erythrocyte membranes, mean platelet volume, and activities of alkaline phosphatse (EC3.1.3.1) and α-d-mannosidase (EC3.2.1.24) in neutrophils did not respond to changes in Zn status. In contrast, alkaline phosphatase activity in erythrocyte membranes showed a significant decline which was consistent in all subjects (nmol product formed/min per mg protein; baselinev.7-week Zn depletion, 0.656 (sd 0.279)v.0.506 (sd 0.230), at 7 weeks;P< 0.05); neutral phosphatase activity remained unchanged. Alkaline phosphatase activity in erythrocyte membranes may be a potential index of Zn status in humans

scholarly journals Zinc status of northern Tasmanian adults

2015 ◽  
Vol 4 ◽  
Author(s):  
Jeffrey M. Beckett ◽  
Madeleine J. Ball
Keyword(s):  
Cross Sectional Study ◽  
Health Concern ◽  
Zn Deficiency ◽  
Dietary Intakes ◽  
Cross Sectional ◽  
Serum Zn ◽  
Zinc Nutrition ◽  
Zn Status ◽  
Low Serum ◽  
Consultative Group

AbstractInformation regarding Zn status in the Australian population is very limited. Mild deficiencies in Zn have been associated with CVD, impaired immune function and poor healing. A cross-sectional study of 497 northern Tasmanian adults (24–82 years of age) was conducted to assess Zn status. Dietary intakes were assessed by FFQ and serum concentrations of Zn were evaluated using International Zinc Nutrition Consultative Group methodology. Mean Zn intakes were 12·6 (sd4·4) mg/d for men and 10·9 (sd3·6) mg/d for women. It was found that 52 % of men but only 9 % of women consumed less than the Australia/New Zealand estimated average requirement for Zn. Mean serum Zn was 13·0 (sd2·4) µmol/l in men and 13·0 (sd2·5) µmol/l in women. Overall, 15 % of men and 7 % of women had low serum Zn levels. Furthermore, low serum Zn was observed in 18 % of men 50 years or older and 30 % of men 70 years or older. The present results suggest that mild Zn deficiency may be prevalent in older Tasmanian adults, particularly men; and due to the importance of Zn in many areas of health, this could be of public health concern.

A rapid real-time qRT-PCR assay for ovine β-actin mRNA

2005 ◽  
Vol 117 (2) ◽  
pp. 173-182 ◽  
Author(s):  
Helga Bjarnadottir ◽  
Jon J. Jonsson
Keyword(s):  
Real Time ◽  
Pcr Assay ◽  
Qrt Pcr ◽  
Actin Mrna
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