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Radioisotopic evidence for the polypeptides associated with photosystem II.

Direct radioisotopic evidence for the polypeptides associated with oxygen evolution in a non-destructive approach by comparing the polypeptides of photosystem II (PS II) particles in etioplasts that lack O2 evolution with that of chloroplasts that exhibit high rates of oxygen evolution is reported. Polypeptide analysis by coomassie blue staining revealed that 32, 23 and 15 kDa proteins were absent in etioplast particles but developed in chloroplast PS II particles after illumination. However, a strikingly different picture was obtained when labelled polypeptides were analyzed from the fluorograph. While the new proteins identified by coomassie blue staining were also labelled, intensely labelled major polypeptides of molecular range 40, 41, 43 kD and minor polypeptides of molecular range 47, 48, 51 kD were observed in chloroplast PS II particles but not in etioplasts or etioplast particles; these labelled polypeptides were so prominent that they could be identified even in thylakoid membranes.

Chlorophyll↗

The determination of lead in environmental air using a radioisotope X-ray-fluorescence unit.

The feasibility for the development of a portable radioisotope excited X-ray fluorescence unit, able to detect Pb in air samples, was studied. The unit employs a 0.6-mCi 109Cd source and a Xe-filled proportional detector. The sensitivity of the method was found to be 15 micrograms of lead per cubic metre of air. Suggestions are also made for the improvement of the sensitivity.

Air Pollutants↗

Objective evaluation of local xenogeneic graft-versus-host reaction by computerized radioisotope imaging (CRI).

The local xenogeneic graft-versus-host reaction (GVHR) assay has been used clinically to evaluate cellular immune competence and experimentally to monitor the immunomodulatory effects of several drugs. By employing a computerized radioisotope imaging (CRI) technique, we were able to perform the assay with smaller numbers of mononuclear cells (MNC) and to rid it entirely of any bias. Measuring the local GVHR by CRI compares well with the conventional measurement of the volume (correlation coefficient r = 0.619; P less than 0.001). A clear-cut distinction was documented between normal donors and cancer patients (P less than 0.001) when 10 X 10(6) or more MNC were used in the assay. This is an improvement over the previous, conventional testing of local GVHR which required injection of 20 X 10(6) MNC in order to achieve a similar resolution. The indications for the presence of immune competence have therefore been redefined using the local GVHR index as determined by CRI according to the scale of MNC injected. Thus, immunocompetence is considered present if the CRI index is greater than or equal to 1.2 for 10 X 10(6) MNC, greater than or equal to 2.0 for 15 X 10(6) MNC and greater than or equal to 2.6 for 20 X 10(6) MNC.

Animals↗

Carbonylmetalloimmunoassay (CMIA) a new type of non-radioisotopic immunoassay. Principles and application to phenobarbital assay.

A new non-radioisotopic immunoassay procedure, which we have termed carbonylmetalloimmunoassay (CMIA), is described. The tracers used in this approach are organometallic carbonyl complexes that can be detected at femtomole levels (300-700 fmol) by Fourier transform infrared (FT-IR) spectroscopy. The validity of the technique has been tested in a phenobarbital assay using as the marker a cyclopentadienylmanganese (I) tricarbonyl (cymantrene) moiety, ethyl acetate extraction to separate the free and bound organometallic fractions, and FT-IR spectroscopy to detect the CO stretching modes of the organometallic label. Typical dilution and standard curves obtained with this CMIA procedure are presented. The method was of comparable sensitivity to a [14C] radioimmunoassay (RIA) for the detection of phenobarbital. A comparison of the results for phenobarbital assays by both CMIA and RIA showed that higher titres were obtained using the CMIA method. The standard curves suggest that CMIA is a reliable and reproducible immunoassay procedure for phenobarbital.

Cross Reactions↗

Activation-induced expression of vascular permeability factor by human peripheral T cells: a non-radioisotopic semiquantitative reverse transcription-polymerase chain reaction assay.

Vascular permeability factor, also known as vascular endothelial growth factor (VPF/VEGF), is a disulfide-linked dimeric glycoprotein of about 40 kDa that enhances vascular permeability, induces chemotaxis and activation of monocytes/macrophages, and promotes growth of vascular endothelial cells. It has been reported that several tumor cell lines and normal cells produce VPF/VEGF. To examine the possibility of VPF/VEGF mRNA expression in human peripheral T cells and its mechanism(s) of regulation, we developed a non-radioisotopic semiquantitative reverse transcription-polymerase chain reaction (RT-PCR; VPF/VEGF, GAPDH co-amplification) assay which detects all species of VPF/VEGF mRNA alternative splicing products. T cells expressed negligible VPF/VEGF mRNA in the resting state. However, TPA markedly enhanced the expression of 121-, 165- and 189-amino-acid-containing isoforms of VPF/VEGF mRNA in T cells. Both CD4+ and CD8+ T cells expressed VPF/VEGF mRNA in response to TPA treatment. Moreover, T cell receptor stimulation with monoclonal anti-CD3 antibody with or without IL-1 beta enhanced VPF/VEGF mRNA expression in T cells. These findings suggest that T cell activation induces VPF/VEGF expression in the cells, resulting in induction of its biological activities.

Endothelial Growth Factors↗

Radioisotope diagnosis of splenic trauma.

Multiple injury or delay in seeking medical attention may prevent confident clinical diagnosis of splenic trauma. The spleen scan is a rapid, simple, noninvasive test useful in such circumstances. When peritoneal lavage is contraindicated, unrevealing, or inapproapriate, radioisotope imaging of the spleen can help confirm a suspicion of splenic injury.

Adolescent↗

Radioisotope angiography in diagnosis of varicocele.

A varicocele, the most common surgically correctable cause of subfertility in men, is a manifestation of abnormal retrograde venous blood flow through the internal spermatic vein into the peritesticular pampiniform plexus. Methods of diagnosing the varicocele, including careful physical examination using a Valsalva maneuver as well as a Doppler stethoscope, remain somewhat subjective. Venography, while a reliable and objective technique, is invasive and not without morbidity. In the present study, 16 patients with clinically detectable varicoceles of various sizes were studied using a testicular radioisotope angiogram developed in our laboratory. Following the injection of 15 mg. of stannous pyrophosphate which results in tagging of the circulating red cell pool, 20 mCi. of 99m-technetium were administered in an intravenous bolus injection. The isotope binds to the pyrophosphate thus tagging the red blood cells and creating a blood-pooling agent with a relatively short half-life (six hours). Perfusion studies as well as static views of the scrotal contents, both in the supine and erect position, with and without Valsalva maneuver, were obtained using a gamma camera. The activity in each hemiscrotum then was quantitated using a digital computer. It is proposed that quantitation of this differential blood pool will make the identification of even subclinical varicoceles a potentially objective determination.

Humans↗

Cost effective analysis of prenephrectomy radioisotope scans in renal cell carcinoma.

To evaluate the cost-effectiveness of liver and bone isotope scans, results of scans were reviewed in 60 patients with renal cell carcinoma. In no patient did the results of the scans influence or change the treatment plan. Routine utilization of radioisotope scans as part of the preoperative evaluation of patients with renal cell carcinoma should be discouraged.

Adenocarcinoma↗

Absorption of irrigating fluid during transurethral prostatic resection as measured by ethanol, radioisotopes, and regular-interval monitoring.

Ethanol monitoring is a novel noninvasive method for immediate detection of absorption of irrigating fluid during transurethral prostatic resection. Its accuracy was evaluated during thirteen resections using 5% mannitol +2% ethanol as the irrigating fluid. The ethanol concentration in the expired breath correlated strongly with the degree of absorption of intravascular irrigating fluid as determined by a radioisotope technique, and fluid volume measurements, and with the changes in the serum sodium concentration. Extravascular absorption of the irrigating fluid resulted in prolonged elevation of the ethanol concentration and gave smaller changes in the serum sodium concentration.

Absorption↗

Radioisotope bone scanning for the detection of occult bony metastases in invasive cervical carcinoma.

Radioisotope bone scans to search for occult skeletal metastases at initial staging were obtained in 540 patients with invasive cervical carcinoma admitted to the Gynecological Oncology Unit of the Tygerberg Hospital over a 36-month period. Positive scans were reported in 55 (10.2%) patients but subsequent radiographic examination revealed that of these, 43 were false positive. The 11 patients with bony metastases were all initially classified as Stage III or IV disease irrespective of the bone scan and the positive scans influenced the clinical staging in only 6 (1.1%) patients. All the bone scans in the 210 patients with clinical Stage I or II disease were negative. Bone scanning does not seem warranted at initial staging in Stage I or II invasive cervical carcinoma.

Bone Neoplasms↗

A non-radioisotopic reverse transcriptase assay using biotin-11-deoxyuridinetriphosphate on primer-immobilized microtiter plates.

We developed a non-radioisotopic (non-RI) reverse transcriptase assay (RTA). The reverse transcriptase (RT) incorporates biotin-11-deoxyuridine-triphosphate (bio-dUTP) using a poly(rA) template hybridized with oligo(dT) primer that is immobilized on the surface of a 96-well microtiter plate. This assay is thus semi-automated by adapting it to an ELISA testing format. The incorporation of bio-dUTP was enhanced by adding cold dTTP to the reaction mixture, optimally in a molar ratio 4:1 (dTTP:bio-dUTP). This non-RI RTA is more sensitive than the conventional RI assay for the detection of purified Rous-associated virus 2 (RAV-2) and of human immunodeficiency virus type 1 (HIV-1) lysate. Because of its simple procedure, higher sensitivity and non-use of RI materials, the assay can be utilized not only for virological studies but also for routine safety screening of biological products for retroviral contamination.

Avian Leukosis Virus↗

Leishmania donovani-macrophage binding mediated by surface glycoproteins/antigens: characterization in vitro by a radioisotopic assay.

A radioisotopic assay was developed to quantitate the binding of Leishmania donovani promastigotes to hamster peritoneal macrophages in vitro. The binding was temperature dependent and required no serum factors. Binding was reduced by preloading macrophages with zymosan granules or unlabeled promastigotes, but not with latex leads or opsonized erythrocytes. Binding was reduced by 10 mM EGTA that was reversible by the addition at an equimolar concentration of calcium, but not magnesium ions. Sialic acid, D-glucose, D-mannose and their derivatives reduced the binding, whereas L-fucose, D-galactose and their related sugars did not. Pretreatment of promastigotes with neuraminidase, alpha-mannosidase, alpha-N-acetylglucosaminidase or beta-glucosidase reduced their binding to macrophages. Prior trypsinization of either macrophages or promastigotes also substantially reduced the binding. At 4 degrees C, prior opsonization of promastigotes with subagglutination titers of antiserum doubled the level of binding but in combination with Protein A reduced it to 50% of its normal binding level. Prior opsonization of macrophages decreased their binding to promastigotes significantly at 4 or 37 degrees C. The results indicate that binding of Leishmania donovani promastigotes to hamster peritoneal macrophages is a ligand-receptor interaction involving their antigenic surface membrane proteins. The binding ligands of the parasites appear to have at least sialol, glucosyl, mannosyl and N-acetylglucosaminyl terminal residues as binding determinants. Thus, receptor-mediated endocytosis, defined in a broader sense, appears to be the mechanism by which leishmanias gain entry into macrophages.

Animals↗

In vitro radioisotope precursor incorporation into ribonucleic acid and protein in dystrophic mouse gastrocnemius muscle (C57 BL/6J dy2J/dy2J).

An in vitro tissue slice technique has been developed and used to compare radioisotope precurosr incorporation into RNA and protein in normal and dystrophic mouse gastrophic muscle. Significant differences are observed with both RNA and protein when incorporation is measured on a fresh weight of muscle basis. Specific activity comparisons show significantly increased incorporation with protein but not with RNA. A comparison of the results with in vivo studies has been made. The in vitro system developed is applicable to the study of macromolecular metabolism in normal and diseased human muscle tissue.

Animals↗

Reversed-phase high-performance liquid chromatography of salmon calcitonin and its degradation products in biological samples using column switching and flow-through radioisotope detection.

For the determination of salmon calcitonin and its degradation products in biological samples, a reversed-phase HPLC method with column switching and flow-through radioisotope detection has been developed using high specific activity [125I]salmon calcitonin. Effects of the precolumn packing material and washing solvent were examined in terms of [125I]salmon calcitonin recovery. Spiked samples of [125I]salmon calcitonin in plasma and kidney homogenate were injected onto a LiChroprep RP-8 precolumn after dilution with 0.1% trifluoroacetic acid. After washing the polar interfering compounds with 0.1% trifluoroacetic acid, the concentrated [125I]salmon calcitonin and its degradation products were eluted and separated on a W-Porex C18 column with a gradient of 0.1% trifluoroacetic acid in acetonitrile-water. Detection and calibration of [125I]salmon calcitonin were possible down to picogram levels. Reproducible kinetic data for the degradation of intact [125I]salmon calcitonin were possible down to picogram levels. Reproducible kinetic data for the degradation of intact [125I]salmon calcitonin by rat kidney homogenate could be traced.

Animals↗

Short- and long-term reproducibility of radioisotopic examination of gastric emptying.

Reproducibility of gastric emptying (GE) of a radiolabelled solid meal was assessed. The short-term reproducibility was evaluated on the basis of 12 paired GE examinations performed 1-3 days apart. Twelve paired GE examinations taken 3-8 months apart enabled long-term reproducibility assessment. Reproducibility of GE parameters was expressed in terms of the coefficient of variation, CV. No significant between-day variation of solid GE was found either regarding the short-term or the long-term reproducibility. Although slightly higher CV values characterized the long-term reproducibility of the GE parameters considered, the variations of the differences between repeated GE examinations did not differ significantly between short- and long-term GE reproducibility. The results obtained justify the use of radioisotopic GE measurement for the assessment of early and late results of pharmacologic or surgical management.

Adult↗

Preliminary results on the production of short-lived radioisotopes with a Plasma Focus device.

An experimental campaign was conducted to assess the feasibility of short-lived radioisotope (SLR) production within the pulsed discharges of a Plasma Focus (PF) device. This so-called "endogenous production" technique rests on the exploitation of nuclear reactions for the creation of SLR directly within the plasma, rather than on irradiating an external target. Until now only one research group has published data relevant to PF endogenous production of SLR, and the data seem to confirm that the PF has the capability to breed SLR. The campaign demonstrated production of (15)O, (17)F and (13)N from the (14)N(d,n)(15)O, (12)C(d,n)(13)N and (16)O(d,n)(17)F reactions. A 7kJ, 17kV Mather-type PF was operated with natural nitrogen, oxygen, CO(2) and deuterium in the vacuum chamber. Results to date confirm that, with a PF of this type, up to 1microCi of SLRs per discharge can be obtained.

Journal Article↗

Accreditation experience of radioisotope metrology laboratory of Argentina.

This work presents the experience developed by the Radioisotope Metrology Laboratory (LMR), of the Argentine National Atomic Energy Commission (CNEA), as result of the accreditation process of the Quality System by ISO 17025 Standard. Considering the LMR as a calibration laboratory, services of secondary activity determinations and calibration of activimeters used in Nuclear Medicine were accredited. A peer review of the (alpha/beta)-gamma coincidence system was also carried out. This work shows in detail the structure of the quality system, the results of the accrediting audit and gives the number of non-conformities detected and of observations made which have all been resolved.

Accreditation↗

Design of dantrolene-derived probes for radioisotope-free photoaffinity labeling of proteins involved in the physiological Ca2+ release from sarcoplasmic reticulum of skeletal muscle.

Bifunctional dantrolene derivatives have been synthesized as probes for radioisotope-free photoaffinity labeling with the aim of elucidating the molecular mechanism of skeletal muscle contraction. GIF-0430 and GIF-0665 are aromatic azido-functionalized derivatives that were designed to selectively inhibit physiological Ca2+ release (PCR) from sarcoplasmic reticulum (SR) in mouse skeletal muscle without a strong effect on Ca2+-induced Ca2+ release (CICR). These photoaffinity probes consist of either an azidomethyl or an ethynyl group, respectively, which could function as a tag for introduction of an optional detectable marker unit by an appropriate chemoselective ligation method after the photo-cross-linking operation. Actually, the former probe worked to photolabel its target proteins specifically as confirmed by subsequent fluorescent visualization.

Animals↗