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[Application of double focusing magnetic sector high resolution ICP-MS to the study of iron stable isotope trace].

Iron absorption efficiency study is one of the key steps to overcome iron deficiency anemia. Determination of iron isotope ratios is the base of iron absorption study. For this purpose radioactive isotope trace method, neutron activation, and thermal ionization mass spectrometry etc. have been used as accessory means. A double focusing inductively coupled plasma mass spectrometry (DF-ICP-MS) was used to determine isotope ratios of Fe in food and human faeces in this paper. Interferences of 40Ar16O to 56Fe and 40Ar14N to 54Fe were avoided by using higher resolution (3000). Longer times and fast electronic scanning were employed to decrease the influence of signal fluctuations from the plasma. Instrumental parameters of the mass spectrometer were optimized while keeping the total integral time under 27s. RSD values obtained for isotope ratios of 5Fe/56Fe, 57Fe/56Fe and 58 Fe/56Fe were 0.23%, 0.14%, 0.22%, respectively, for a natural standard solution of 5 microg x mL(-1). The method has been used to iron nutrition studies.

Iron↗

Neonatal water metabolism: an objective postnatal index of intrauterine fetal growth.

The water metabolism of 103 newborn babies was determined over the first 10 postnatal days, by measuring water turnover rates by means of an isotope dilution technique. This technique involves the oral administration of the non-radioactive isotope of water, 2H2O, and the measurement of its urinary excretion by infrared spectrophotometry. The slope of the excretion curve after equilibration with the infant's body water was mathematically expressed as the rate constant. Using multiple obstetric and paediatric criteria, the babies were clinically classified into one of three categories, fully grown ("normal'), borderline or clearly growth retarded. The median values of the rate constants X 10(4) (h-1) for the three groups were 73.3, 85.9 and 100.2 and were highly significantly different from each other (P less than 0.0005) with no overlap of the 97% non-parametric confidence limits of each group. Neonatal water turnover increased with the clinical degree of intrauterine fetal growth retardation and within the limits of this study, this finding was unaffected by gestational age, birth weight or the neonatal environment. The results suggest that neonatal water metabolism is an objective postnatal index of fetal growth retardation.

Body Water↗

A novel method for sentinel lymph node mapping using magnetite in patients with non-small cell lung cancer.

OBJECTIVE: The detection rate of sentinel lymph nodes in patients with non-small cell lung cancer using isosulfan blue dye is too low for clinical use. Although exposure to radioactivity is reportedly minimal, special procedures are nonetheless required when a radioactive isotope is used as a tracer. Therefore, to eliminate the need for a radioactive tracer and to obtain a better detection rate than is obtained with isosulfan blue dye, we have developed a novel method that employs magnetite as the tracer. The aim of the present study was to test the feasibility of this technique. METHODS: The tracer employed was ferumoxides, a colloidal superparamagnetic iron oxide of nonstoichiometric magnetite. Thirty-eight non-small cell lung cancer patients participated in the study; each received 5 mL of ferumoxides, injected around the tumor intraoperatively. Fifteen minutes after injection, lung resection and lymph node dissection were carried out. The magnetic force within the lymph nodes was measured using a highly sensitive handheld magnetometer ex vivo. All lymph nodes were also subjected to conventional histological analysis. RESULTS: The rate of detection of sentinel lymph nodes was 81.6% (31/38). The accuracy, sensitivity, and false-negative rates were 96.8% (30/31), 85.7% (6/7), and 14.3% (1/7), respectively. CONCLUSION: Intraoperative sentinel lymph node mapping using ferumoxides and a highly sensitive magnetometer is a safe, accurate, and sensitive way to detect sentinel lymph nodes in non-small cell lung cancer patients.

Adult↗

[A novel method for sentinel lymph node mapping using magnetite].

The detection rate of sentinel lymph nodes using isosulfan blue dye is too low for clinical use. Although exposure to radioactivity is reportedly minimal, special procedures are nonetheless required when a radioactive isotope is used as a tracer. Therefore, to eliminate the need for a radioactive tracer and to obtain a better detection rate than is currently obtained with isosulfan blue dye, we have developed a novel method that employs magnetite as the tracer. The tracer employed is ferumoxides, a colloidal superparamagnetic iron oxide of non-stoichiometric magnetite. The magnetic force within the lymph nodes was measured using a highly sensitive hand-held magnetometer ex vivo. The rate of detection of sentinel lymph nodes was 82% in non-small cell lung cancer. Intraoperative sentinel lymph node mapping using ferumoxides and a highly sensitive magnetometer is a safe, accurate, and sensitive way to detect sentinel lymph nodes. To detect sentinel lymph nodes in vivo, the magnetometer should be sterilized and the sensitivity of the magnetometer should be higher than that we used. Therefore now we are developing a more sensitive and sterilizable magnetometer.

Contrast Media↗

Detection of p53 mutations by single-strand conformation polymorphisms (SSCP) gel electrophoresis. A comparative study of radioactive and nonradioactive silver-stained SSCP analysis.

p53 mutations are the most common genetic abnormality in humans tumors, but their clinical significance remains to be precisely elucidated. Conventional single-strand conformation polymorphism (SSCP) analysis, a well-established technique for detecting p53 mutations, uses radioactively labeled polymerase chain reaction (PCR) products, which migrate abnormally in the presence of mutations. We performed radioactive PCR-SSCP analysis in a series of 30 formalin-fixed, paraffin-embedded ovarian carcinomas and two cell lines (SW480 and Caov4) harboring known homozygous p53 mutations and compared the results with nonradioactive silver-stained SSCP. The purpose was to assess whether nonradioactive SSCP is suitable for detecting p53 mutations in a rapid, sensitive, cost-effective fashion, without the need of radioactive isotopes. We accomplished PCR amplification of p53 exons 5 through 8 in 26 carcinomas, and radioactive SSCP detected p53 mutations in 13 tumors; three mutations were localized in exon 5, six in exon 6, two in exon 7, and two in exon 8. All mutations were correctly identified with nonradioactive SSCP, except for one exon 8 mutation. To establish the sensitivity of nonradioactive SSCP, DNA samples of SW480 and Caov4 were mixed with increasing amounts (0-90%) of normal DNA and subjected to PCR-SSCP analysis. Mutations were detected until the concentration of SW480 and Caov4 was 15% and 10%, respectively, of the total sample. The results of our investigation demonstrate that nonradioactive silver-stained SSCP is a sensitive, rapid, and simple technique to detect p53 mutations, even in formalin-fixed tissues, and could be easily used to investigate large series of patients to assess the clinical significance of p53 mutations in human tumors.

Carcinoma↗

One step direct detection of recurrent mutations in the breast cancer susceptibility gene, BRCA1.

Germ-line mutations of the BRCA1 gene have been linked to 85% of hereditary breast and ovarian cancers. More than one hundred mutations have been reported, including several which are over represented within the Ashkenazi Jewish (185delAG) and Irish families (1294del40). These recurrent mutations are cost-effective targets for presymptomatic screening of cancer susceptibility. Most current techniques such as single strand conformation polymorphism, heteroduplex analysis, DNA sequencing, and protein translation termination assays are multistep, time consuming methods and require radioactive isotopes for mutation detection. As an alternative, we have developed a single step, non-radioactive allele specific oligonucleotide (ASO) PCR assay designed to target any known mutation. Its application to detection of the BRCA1 185delAG and 1294del40 mutations is described here. The ASO PCR is efficient, highly sensitive, non-radioactive, specific for individual mutations, performed in a single step, and is amenable to large-scale screening for other known mutations.

Alleles↗

[Radionuclide detection of Helicobacter pylori infection].

The aim is the assessment of the HP infection in stomach using breath test and comparison to other diagnostic methods, as well as following up the effect of therapy. In 83 patients with digestive discomfort rapid urease test, histology and breath test were performed, while in 25 patients with proven HP infection the effect of therapy was followed up using breath test and clinical findings. For rapid urease test and histology, samples were taken from antral mucosa. Breath test was performed after per oral administration of the capsule of 14C- urea (37 kBq) (Izotop, Hungary and Laboratory for radioactive isotopes, Vinca) which, in the presence of Helicobacter pylori breaks up to 14CO2 and NH3. Radioactivity was measured by beta counter in the exhaled air fasting and 30 minutes after ingestion of the capsule. According to our results, the rise of activity over 100% was considered positive. From 83 patients, 58 were breath test was positive, 24 negative and one equivocal. Fast urease test was in 54 positive, in 29 negative while histology was in 57 postitive and 26 negative. Findings of the breath and urease tests were in accordance in 93% patients while breath test and histology in 98% patients. During follow up of the therapeutic effects, breath test and clinical findings were in accordance in 98% patients. Breath test can be useful in diagnosis but is a method of choice in following up the patients after therapy for H. pylori infection, because it is non-invasive, fast and precise.

Breath Tests↗

Extraction of caesium from mosses and incorporation into an oxalate crystalline lattice.

The ratios of stable and radioactive isotopes in the environment are not constant and their effects on living organisms are not the same. Whilst radionuclides can be monitored by measuring their radioactivity levels, it is also important to determine the stable isotopes. The method described in this paper represents a simple and cheap process by which to extract caesium from moss, without significant destruction of the plant, and to incorporate the isolated caesium into a water-soluble crystal. Ammonium oxalate and phosphoric acid are water-soluble substances at 5% concentration but, when the solutions are combined, crystallisation occurs. Crystals were estimated to contain 68.30% O, 20.98% C, 4.68% H, 6.04% N and less than 0.01% P, corresponding to NH4HC2O4 x H2C2O4 x 2H2O (C4H11NO10). The crystal mass did not vary with the species of moss, although the effectiveness of caesium extraction did. Typically, 73.0 +/- 6.1% of the solubilised caesium was entrapped within the crystals.

Bryophyta↗

An in vitro system for measuring intrinsic dietary mineral exchangeability: alternative to intrinsic isotopic labeling.

The primary purpose for the in vitro system described here was to provide an alternative method to intrinsic isotopic labeling for determining exchangeability of intrinsic food mineral with extrinsic inorganic mineral or one of its isotopes. In this system, foods or food mixtures extrinsically tagged with an isotope of the mineral of interest are incubated successively in media simulating peptic or pancreatic digestion. Radioactivity and mineral measurements are made on a)the mixture before digestion and after b)peptic and c)successive peptic pancreatic (P-Pa) digestion. Exchangeability is determined by comparing the specific activity in the peptic and P-Pa digest supernatants to that in the mixture before digestion. In addition, determination of the total soluble mineral after P-Pa digestion provides a measurement of potential bioavailability. The procedure was tested for calcium (Ca) exchangeability and bioavailability from a number of foods and food combinations with 45Ca as the tracer. It could be used for any mineral, however, including non-nutrient or toxic elements. It is likely to be most useful in association with human studies carried out with extrinsic stable (non-radioactive) or short-lived radioactive isotopes that are unsuitable for intrinsic labeling.

Biological Availability↗

[Applications of stable isotopes in biomedical sciences].

The employment of stable isotopes technology allows to perform a wide range of studies in vivo avoiding the utilisation of radioactive isotopes. As stable isotopes naturally occur in nature, the technique detects the enrichment in breath air of the tracer administered. Stable isotopes technology is increasingly being used in biomedical investigation due to the security of its use and the development and availability of new substrates. Breath tests such 13C-urea in the detection of Helicobacter pylori and 13C-octanoic acid for gastric emptying are the most important clinical applications.

Amino Acids↗

Pharmacokinetics of entacapone, a peripherally acting catechol-O-methyltransferase inhibitor, in man. A study using a stable isotope techique.

OBJECTIVE: This study investigated the pharmacokinetics of the catechol-O-methyltransferase (COMT) inhibitor entacapone by giving simultaneously stable non-radioactive isotope 13C-entacapone intravenously (i.v.) and unlabelled entacapone orally. In comparison with a crossover design, the simultaneous i.v. and oral administration made it possible to minimise intra-individual variation, sample size and the duration of the study and still obtain accurate pharmacokinetic data. METHODS: Eight healthy male volunteers were enrolled in this study. They were given a 20-mg i.v. dose of 13C-entacapone as a 1-mg/ml infusion at a constant rate of 5 mg/min over 4 min and a 100-mg dose of unlabelled entacapone orally immediately after the infusion. Blood samples were drawn at -5 (before onset of infusion), 0 (upon termination of infusion), 2, 5, 10, 20, 30 and 45 min and 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 10 and 12 h after the tablet ingestion. Urine during the 48 h after dosing was collected in fractions. Concentrations of 13C-entacapone and entacapone in plasma samples and urine fractions were determined using gas chromatography-mass spectrometry. RESULTS: The decay of i.v. 13C-entacapone in plasma was tri-exponential and its pharmacokinetics were described using an open three-compartment model. The volume of the central compartment (Vc) and the volume of distribution at steady state (Vss) were 0.08+/-0.03 l/kg and 0.27+/-0.10 l/kg, respectively. Total plasma clearance (Cltot) averaged 11.7+/-1.9 ml/min kg(-1). The half-lives for the distribution phase and for the rapid and terminal elimination phases (t1/2alpha, t1/2beta and t1/2gamma) were 0.05+/-0.01 h, 0.38+/-0.16 h and 2.40+/-1.70 h, respectively. The terminal elimination phase accounted for only 9% of the total area under the plasma concentration-time curve (AUC), which was 409 +/- 98 ng h/ml after the i.v. dose. Oral entacapone was absorbed rapidly with a time to reach the peak concentration (tmax) of 0.9+/-0.4 h, a maximum concentration (Cmax) of 457+/-334 ng/ml and an AUC of 497+/-118 ng h/ml. During the 48 h after dosing, the recovery of free and conjugated unchanged 13C-entacapone in urine was 38.1+/-7.2% of the i.v. dose and the recovery of free and conjugated unchanged entacapone 13.3+/-3.9% of the oral dose. The bioavailability of oral entacapone was 25% based on the AUC values and 35% based on urinary excretion. CONCLUSION: The results of the present study using stable isotope technique indicate that entacapone is rapidly absorbed, distributed to a small volume and rapidly eliminated by mainly non-renal routes. The pharmacokinetic profile of entacapone provides the rationale for a concomitant and frequently repeated simultaneous dosing of entacapone with levodopa and dopa decarboxylase inhibitors in the treatment of Parkinson's disease. This study confirmed the previously published data and fully support the validity of the technique used.

Administration, Oral↗

[Automated evaluation and analysis of isotopic heart function tests with minimal cardiac transit times].

The clinical evaluation of cardiac function by non-invasive means with the help of radioactive isotopes registering minimal cardiac transit times (MTTs) is increasingly employed in routine diagnosis. In order to economise on work load and time expenditure and for the purpose of an objective and complete evaluation of data, automatic data processing is desirable. This paper describes a program that consists of 4 sections: 1. examination 2. generation of data 3. processing of data 4. evaluation of data. The program permits a nearly total automated data generation, analysis, calculation, classification, final clinical evaluation and the automated production of a medical report. This greatly reduces the time spent per examination to approximately 5 minutes. The automated evaluation of the data is based on clinical experience with approximately 3.500 measurements in patients with the most frequent cardiac diseases. The result is an objective statement of causes of MTT changes and is highly useful for medical routine application. The control and the final inclusion of the findings into the spectrum of other clinical results remains the subject of the physician's judgement. The advantage of the method is the evaluation of all cardiac segments. This pertains to the evaluation of both atria and both ventricles, as well as of the entire central circulation with and without participation of the lung.

Computers↗

Induction of stress proteins by electromagnetic fields in cultured HL-60 cells.

HL-60 cells in culture were exposed for 2 h to a sinusoidal 0.1 or 1 mT (1 or 10 Gauss) magnetic field at 60 Hz and pulse labeled after exposure with radioactive isotopes by incubation by using either [(35)S]methionine, [(3)H]leucine, or [(33)P]phosphate. The radioactive labels were incorporated into cellular proteins through synthesis or phosphorylation. Proteins were extracted from electrostatically sorted nuclei, and the heat shock/stress proteins (sp) were analyzed for synthesis and phosphorylation by two-dimensional polyacrylamide gel electrophoresis. In the control cultures (no exposure to the magnetic field), sp 72c (cognate form) was faintly observed. A 0.1 mT exposure did not show sp metabolism to be different from that of the controls; however, after a 1 mT exposure of the HL-60 cells, sp 70i (inducible form) was synthesized ([(35)S]methionine incorporation). Sp 90 was not synthesized at either field level, but was phosphorylated ([(33)P]phosphate incorporation) in the 1 mT exposure. Sp 27 (isoforms a and b) was induced after a 1 mT exposure as reflected by labeling with [(3)H]leucine. These sps were not detected after a 0.1 mT exposure. After a 1 mT exposure and labeling with [(33)P], sp 27 isoforms b and c were phosphorylated whereas isoform 'a' was not observed. Sps 70i, 72c, and 90 were identified by commercial sp antibodies. Likewise, polypeptides a, b, and c were verified as sp 27 isoforms by Western blotting. Statistical evaluation of sp areas and densities, determined from fluorographs by Western-blot analysis, revealed a significant increase in sps 90 and 27a after a 1 mT magnetic field exposure. The 1 mT magnetic field interacts at the cellular level to induce a variety of sp species. Bioelectromagnetics 20:347-357, 1999. Published 1999 Wiley-Liss, Inc.

Electromagnetic Fields↗

Disposal of low-level radioactive waste. Impact on the medical profession.

During 1985, low-level radioactive waste disposal has become a critical concern. The issue has been forced by the threatened closure of the three commercial disposal sites. The medical community has used radioactive isotopes for decades in nuclear medicine, radiation therapy, radioimmunoassay, and biomedical research. Loss of disposal capacity for radioactive wastes generated by these activities, by the suppliers of radioisotopes, and by pharmaceutical companies will have a profound impact on the medical profession.

Legislation as Topic↗

Prostate brachytherapy. An overview.

BACKGROUND: Prostate brachytherapy represents one of the oldest techniques of using radiation therapy to treat prostate cancer. Over the past 10 years, there have been major changes in the types of prostate brachytherapy that can be performed with the introduction of new radioactive isotopes, new afterloading techniques, and an improved understanding of the radiobiology associated with differing dose rates. METHODS: Prostate brachytherapy can be divided into temporary implantation using high activity sources such as iridium-192, or permanent brachytherapy using the interstitial implantation of iodine-125 or palladium-103 sources. There are various techniques that can be used to actually insert the radioactive material into the prostate. This can be done as an open or closed procedure and can be performed via a suprapubic or a perineal retropubic approach. The use of remote afterloading has substantially reduced the radiation protection problems associated with manually loaded radioactive sources. RESULTS: Results using brachytherapeutic isotopes in the treatment of prostate cancer have been variable, but it appears that using the higher dose rate sources and delivering a high relative integral dose to the prostate can result in improved histologic control of prostate cancer. In patients with aggressive prostate cancer, the use of iodine-125 permanent implantation has not been successful. CONCLUSIONS: The role of brachytherapy in the treatment of prostate cancer remains an exciting alternative in the management of prostate cancer. Its role is becoming more defined in the treatment of large, bulky prostatic neoplasms as a way of improving the dose distribution achieved between normal and tumor tissue.

Brachytherapy↗

Tc99m macroaggregated albumin cisternography for the detection of persistent cerebrospinal fluid (CSF) leak in an animal model.

The present study describes a new in vivo animal model that enables the detection of cerebrospinal fluid (CSF) leakage after dural injury. A polyethylene catheter (PE 10) was inserted into the subarachnoid space in the lumbar area by a simple surgical procedure and a radioactive isotope Tc99m Macroaggregated Albumin (Tc99m MAA) was injected into the CSF. In the experimental group, a standardised dural puncture was performed in the cervical area. The accumulation of the isotope in the gauze placed over the dural puncture and viewed by a gamma camera as a spot of concentrated radioactivity, was indicative of CSF leakage. In a second group of animals with intact cervical dura the absence of leakage was presented as a picture of sporadic background radioactivity. To demonstrate the effectiveness of the model in detection of invisible leakage, blood was applied over the cervical dural defect in another group of animals and CSF leakage was assessed by the above mentioned isotope detection method. This in vivo model may be used for evaluation of the sealing properties of various materials under physiological and metabolic processes in living tissue.

Animals↗

A modified technique allowing interactive ultrasound-guided three-dimensional transperineal prostate implantation.

PURPOSE: Ultrasound-guided transperineal prostate implantation is a new technique for performing permanent isotope implants of the prostate. The details of the technique are presented to demonstrate its ability to place radioactive seeds three-dimensionally within the prostate gland to achieve uniform dose distribution without the need for complicated preplanning. METHODS AND MATERIALS: An accurate measurement of the prostate volume is made using biplanar transrectal ultrasound. The total activity to be implanted is derived from a look-up table based on prostate volume. The basic plan is to implant 60-70% of the total activity in the periphery of the gland and the remaining activity in the interior of the gland. The ultrasound transducer provides visualization of the prostate through transverse, longitudinal and oblique cuts and allows for accurate placement of implant needles, approximately 1 cm apart. In addition, these needles can be moved through the prostate under constant visualization, thus allowing for precise seed placement. RESULTS: The setup of the transrectal ultrasound device as well as prostate volume measurements are performed in 10 to 15 min. The actual placement of the needles and seed implantation takes 1 to 1.5 h to perform. Postimplantation dosimetric evaluation is performed using orthogonal x-ray films and 3 mm thick CT slices taken at 3 mm intervals. This evaluation has confirmed accurate seed placement within the prostate gland. CONCLUSION: Interactive ultrasound guided transperineal prostate implantation is a fast and accurate method of performing permanent radioactive isotope prostate implants.

Brachytherapy↗

Yeast tRNA Leu UAG. Purification, properties and determination of the nucleotide sequence by radioactive derivative methods.

A second major species of leucine tRNA, tRNA Leu UAG (formerly designated tRNA Leu CUA) was purified from baker's yeast in a three-step procedure entailing BD-cellulose chromatography in the presence and absence of Mg2+ and Sephadex G-100 gel filtration. Results of aminoacylation and partial RNase T1 digestion experiments showed that this tRNA retains a native conformation under conditions that denature yeast tRNA Leu m5CAA (tRNA3 Leu). The primary structure of baker's yeast tRNA Leu UAG was elucidated by application of sensitive radioactive isotope derivative ("postlabeling") methods. Complete RNase T1 and A and partial RNase U2 fragments, prepared from non-radioactive tRNA and 5'-half and 3'-half molecules, were separated by two-dimensional polyethyleneimine-cellulose anion-exchange thin-layer chromatography and isolated by a novel micropreparative procedure affording high yields of these compounds in sufficient purity for subsequent tritium derivative analysis. Base composition and sequence of oligonucleotides were analyzed by tritium derivative methods. Molar ratios of the fragments were determined from the radioactivity of 3H-labeled nucleoside trialcohols in combination with base analysis. 2'-O-Methylated guanosine was characterized using the [gamma-32P]ATP/polynucleotide kinase reaction. The analysis of classical complete and partial RNase digests by the tritium derivative methods yielded the complete nucleotide sequence of the tRNA. A total of about 20 A260 units of the RNA was used for analysis, i.e. considerably less material than required for conventional spectrophotometric analysis. A different sequencing approach, consisting of a combination of "readout sequencing" with tritium sequencing of complete RNase T1 and A fragments, was applied to the 3'-half molecule. The 3'-half molecule was labeled with 32P at its 5' terminus, partially degraded with RNase T1, U2, and Phy1 and with alkali, and subjected to polyacrylamide gel electrophoresis. The sequence was read off the gel on the basis of cleavage patterns and size of the fragments. While the readout procedure provided only the positions of A, U, C, and G residues in the chain, additional information from tritium derivative analysis was utilized to define the positions of the modified nucleosides. The readout sequencing procedure was found to require less than 0.01 A260 unit of RNA and the analysis of the complete fragments about 6 A260 units. Interesting structural features of tRNA Leu UAG are (a) the location of unique, leucine tRNA iso-acceptor-specific sequences next to U-8, a constant nucleotide participating in synthetase recognition, (b) the occurrence of 1-methyladenosine in the T loop, a modification not present in the structurally related tRNA Leu m5CAA, and (c) the unusual presence of an unmodified uridine in the first position of the anticodon, which may be related to the unusual coding properties reported for this tRNA.

Adenine↗