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Biological damage from the Auger effect, possible benefits.

Decay of radioactive isotopes by K-capture leads to the Auger effect and results in the loss of several orbital electrons and the emission of X-rays. Whereas radiation effects are produced from the emitted electrons, the consequences of the Auger effect are strictly localized to the site of the decaying nuclide. The paper reviews the biological consequences of the decay of 125I which produces the Auger effect. Nearly all data were obtained from DNA labeled with 125I-5-iodo-2'-deoxyuridine (IUdR) in bacteria and mammalian cells. Parameters of effects were cell death, DNA strand breaks, and mutation induction. In order to recognize in a cell the contribution from the Auger effect and that of absorbed radiation, experimental data are analysed in terms of the specific energy for the nuclear volume which contains the isotope. The data indicate that decay of 125I is far more toxic than is expected on the basis of absorbed dose to the labeled nucleus. Moreover, it is emphasized that the toxicity of the 125I decay is largely determined by events immediately localized to the site of decay. Because the consequences of the Auger effect are stricly localized to the molecular site of the decay, 125I and perhaps other nuclides decaying by K-capture promise to be interesting tools in cell biology and molecular biology. First data on the Auger effect as a tool are summarized. It appears that recognizable biological damage is only observed when the Auger effect takes place in vitally important molecules, an example of which is DNA.

Cell Line↗

Applications of nuclear medicine in ophthalmology.

Diagnostic techniques using radioactive isotopes can be used for the investigation of cerebral, orbital and intraocular tumours and for the dynamic study of tear flow. The value of such techniques in the study of obstruction of the nasolacrymal system (particularly partial and functional obstructions) has been established and the possibility of using Tc99m clearance for the investigation of dry eyes is being investigated.

Brain↗

On problems of calculating energy expenditure and substrate utilization from respiratory exchange data.

Indirect calorimetry based on respiratory exchange measurement has been successfully used from the beginning of the century to obtain an estimate of heat production (energy expenditure) in human subjects and animals. The errors inherent to this classical technique can stem from various sources: 1) model of calculation and assumptions, 2) calorimetric factors used, 3) technical factors and 4) human factors. The physiological and biochemical factors influencing the interpretation of calorimetric data include a change in the size of the bicarbonate and urea pools and the accumulation or loss (via breath, urine or sweat) of intermediary metabolites (gluconeogenesis, ketogenesis). More recently, respiratory gas exchange data have been used to estimate substrate utilization rates in various physiological and metabolic situations (fasting, post-prandial state, etc.). It should be recalled that indirect calorimetry provides an index of overall substrate disappearance rates. This is incorrectly assumed to be equivalent to substrate "oxidation" rates. Unfortunately, there is no adequate golden standard to validate whole body substrate "oxidation" rates, and this contrasts to the "validation" of heat production by indirect calorimetry, through use of direct calorimetry under strict thermal equilibrium conditions. Tracer techniques using stable (or radioactive) isotopes, represent an independent way of assessing substrate utilization rates. When carbohydrate metabolism is measured with both techniques, indirect calorimetry generally provides consistent glucose "oxidation" rates as compared to isotopic tracers, but only when certain metabolic processes (such as gluconeogenesis and lipogenesis) are minimal or / and when the respiratory quotients are not at the extreme of the physiological range. However, it is believed that the tracer techniques underestimate true glucose "oxidation" rates due to the failure to account for glycogenolysis in the tissue storing glucose, since this escapes the systemic circulation. A major advantage of isotopic techniques is that they are able to estimate (given certain assumptions) various metabolic processes (such as gluconeogenesis) in a noninvasive way. Furthermore when, in addition to the 3 macronutrients, a fourth substrate is administered (such as ethanol), isotopic quantification of substrate "oxidation" allows one to eliminate the inherent assumptions made by indirect calorimetry. In conclusion, isotopic tracers techniques and indirect calorimetry should be considered as complementary techniques, in particular since the tracer techniques require the measurement of carbon dioxide production obtained by indirect calorimetry. However, it should be kept in mind that the assessment of substrate oxidation by indirect calorimetry may involve large errors in particular over a short period of time. By indirect calorimetry, energy expenditure (heat production) is calculated with substantially less error than substrate oxidation rates.

Animals↗

The use of ionophores of rapid loading of human red cells with radioactive cations for cation-pump studies.

Techniques are described for the rapid loading of intact human red cells with radioactive isotopes of alkali cations or Ca2+ by using ionophorous compounds (nigericin, gramicidin D and A 23187). Loading was rapid and efficient if the membrane potential of the cells was rendered more negative inside. After cation loading the ionophores could be bound to albumin and removed by repeated washings. The ATP and 2,3-DPG contents of the cells were practically unaltered by this treatment. Passive membrane permeability to Na+ and Ca2+ returned to normal. Loaded erythrocytes pumped out Na+ in a ouabain-sensitive and Ca2+ in a lanthanum-sensitive way. Ca2+ -loaded red cells were microspherocytes and exhibited a rapid K+ -efflux. Parallel with the extrusion of Ca2+ cells regained their biconcave shape and normal passive permeability to K+.

Adenosine Triphosphate↗

The impact of W. K. Röntgen's discovery on the use of internalizable sources of ionizing energy in diagnostic and therapeutic nuclear medicine.

The early history of cathode rays, X-rays and a third kind of natural radiation from several minerals and atomic fission is described. In this way the fundamental concept of radioactivity, laws of decay and atomic models were developed. With artificial radioactive isotopes, new tailored radiopharmaceuticals could be introduced into metabolic research, medical diagnostics and therapy. Von Hevesy's concept of the dynamic state of body constituents led to examination of the locations and movements of labelled atoms and molecules as a function of time. That was the birth of nuclear medicine. The principles and value at the molecular level of several specific tracer studies in research and diagnostic or therapeutic use are explained. Typically, diagnostic tests with tracer agents are non-invasive and have low radiation exposure. Competing with other diagnostic and therapeutic modalities, nuclear medicine is a speciality in its own right. But there are moves to classify it as a subspecialization of other organ-oriented clinical disciplines. That is a misunderstanding of the radiologist's role and does not answer the question: What is the best way of working for the patient? New horizons in diagnostic modalities, biochemistry, immunology, imaging and the use of immunogenic therapeutic agents demand a continuous cooperation within interdisciplinary teams. That is as necessary with radiologic departments, participating in changed organizational structures, as with other clinical departments.

Humans↗

Complementarity of magnetic resonance spectroscopy, positron emission tomography and single photon emission tomography for the in vivo investigation of human cardiac metabolism and neurotransmission.

The three techniques allowing the noninvasive study of cardiac metabolism, namely magnetic resonance spectroscopy (MRS), positron emission tomography (PET) and single photon emission computed tomography (SPET), all use external detection with stable or radioactive isotopes. These techniques yield different information. PET is quantitative and very sensitive, and therefore only tracer amounts of molecules need to be injected. It allows neurotransmitters and receptors to be studied and a global view of metabolism (oxygen consumption, glucose and fatty acid utilization) to be obtained. SPET also has good sensitivity, but uses gamma-emitting isotopes of heteroatoms. Their longer half-lives allow follow-up for hours or days. MRS is based on stable elements with high (hydrogen 1, phosphorus 31, fluorine 19...) or low (carbon 13, Deuterium) natural abundance. It has very low sensitivity and only millimolar concentrations of substrates can be detected, but various parts of metabolism can be studied. The in vivo measurement of myocardial concentration of substances has many problems that are common to all three techniques (measurement of the volume, measurement of the quantity of each molecule, resolution, partial volume effect, improvement of the signal-to-noise ratio, movement of the organ). The complementarity of the techniques is illustrated by their applications to the study of cardiac metabolism. For instance, the energy metabolism can be studied by 31P-MRS, which detects the high-energy compounds ATP and phosphocreatine, and 13C-MRS yields information on the tricarboxylic acid cycle activity. PET and SPET allow the utilization of fatty acids, the normal fuels of the heart, to be studied. During ischaemia, PET with 18F-fluorodeoxyglucose (18FDG) can determine the glucose consumption and 1H-MRS shows the increase in lactic acid, reflecting anaerobic glycolysis. Comparison of the use of acetate labelled with 11C for PET or 13C for MRS shows the potentials and limitations of each technique. Myocardial perfusion can be evaluated directly with various PET tracers or indirectly with thallium 201 or various technetium-99m-labelled tracers by SPET. No MRS marker of perfusion is so far clinically available. Mainly SPET and PET are used clinically for the investigation of ischaemic heart disease as well as cardiomyopathies, but some initial results using 31P-MRS are being obtained.

Animals↗

Use of a DNA microfluorometric assay to measure proliferative response of mink lung cells to purified TGF beta and to TGF beta activity found in prostate cell conditioned medium.

The proliferative response of Mv1Lu cells to purified TGF beta 1, or TGF beta-like activity released by various cells into medium conditioned over a 24-h period was quantitated by adapting a rapid DNA fluorometric assay. Acid activation of the conditioned medium allowed the amount of biologically latent versus active TGF beta to be quantitated. A neutralizing antibody specific for TGF beta 1, 1.2, and 2.0 completely blocked the growth inhibition observed treating Mv1Lu cells with either purified TGF beta 1 or medium containing secreted TGF beta-like activity conditioned by DU145 prostate cells. In contrast to other assays commonly used to measure TGF beta activity, the proliferative response is related directly to DNA content rather than as a reflection of enzymatic activity or incorporation of 3H-thymidine. The necessity for radioactive isotope usage has been eliminated, and the biological response can be quantitated over a period of days.

Animals↗

Cultured human skin fibroblasts absorb 65Zn. Optimization of the method and study of the mechanisms involved.

The radioactive isotope 65Zn was used to study the incorporation of zinc by cultured human skin fibroblasts. The development of the method for studying cell uptake of 65Zn in a minimal synthetic medium is presented. Kinetics carried out on control cultures up to 240 min indicated that zinc uptake occurred in three phases, the first being the most rapid. Temperature and pH affect zinc uptake, in favor of an active transport process. In addition, the rate of incorporation is considerably decreased during the first phases after adding potassium cyanide, during the last phases after adding sodium iodoacetate, and during all the phases if dithioerythritol is used. A hypothesis is therefore proposed according to which several types of mechanisms would be involved in zinc uptake by fibroblasts. At least a part of these mechanisms is energy-dependent.

Adult↗

The changes of protein kinase C activity in peripheral blood lymphocytes in the patients with obstructive jaundice and the implication.

The roles of protein kinase C (PKC) signal pathway in the pathogenesis of obstructive jaundice were studied. PKC from cytosolic and membrane fractions of peripheral blood lymphocytes (PBL) in 51 patients with obstructive jaundice and 16 cases of normal controls was isolated and purified. The activities of PKC were determined by radioactive isotope gamma-32P-ATP-catalyzing assay. The results showed that the total PKC activities in PBL in the patients with obstructive jaundice were significantly increased as compared with those in the normal controls (P < 0.01). Moreover, the membrane PKC activities and their percentages of the total PKC activities were higher in obstructive jaundice group than in those in the normal controls (P < 0.05). The total PKC activities in PBL in the patients with obstructive jaundice were significantly positively correlated with the levels of soluble IL-2 receptor (sIL-2R) (r = 0.58, P < 0.01) and the degree of jaundice (T-BIL) (r = 0.67, P < 0.01) in serum. It was concluded that the activities of PKC signal pathway was related with the degree of T-BIL. PKC signal pathway might took part in the activation of T-lymphocytes in the patients with obstructive jaundice and play an important role in the immune regulation and the assessment of pathosis in the patients with obstructive jaundice.

Adult↗

Chernobyl radioactivity on the Black Sea coast of Turkey.

After the Chernobyl reactor accident, Eastern Black Sea coast was one of the heavily contaminated regions of Turkey. Clouds loaded with radioactive isotopes arrived the region on May 1986 and emptied their contents with the heavy rains that are frequently seen in the region. In order to asses the current level of contamination, several different samples, moss, lichen, litter, surface soil and soil cores were collected on August 1994. Samples were brought to the laboratory and their moisture, pH and organic matter contents were determined. Gamma-ray spectra of the samples were collected with a HpGe detector. (137)Cs was the major isotope observed. Activity of most litter samples were below 1000 Bq/kg, while most of the moss samples had activities below 5000 Bq/kg, there were a few with higher (137)Cs activities. Surface soil samples generally had activities less than 2000 Bq/kg and depth profiles of cesium activities in the soil cores showed regional variations.

Journal Article↗

Use of monoclonal antibodies in colorectal cancer: a review.

This is a review article covering the development of monoclonal antibodies attached to a radioactive isotope which are used in scanning for metastatic adenocarcinoma of the colon. The most recent and complete study described 223 patients, of which 169 were evaluated. All patients required an operation. At operation, biopsies were taken of the resected tumor. Some false negative areas, such as dilated varicosities and inflammatory areas, were encountered. The latter can be seen as a chronic inflammatory process such as a leaking anastomosis with a diverticulum or old traumas in the pelvis. This technique allows a physician to follow patients, especially those who have had an abdominal perineal or exoneration of their pelvic organs, for possible recurrence in this area. There is also some discussion about possible future uses in the treatment of the metastatic disease.

Adenocarcinoma↗

Yttrium90 synovectomy in the management of chronic knee arthritis: a single institution experience.

Radiation synovectomy (RS) has been used to treat chronically inflamed joints refractory to treatment using conventional agents. In RS, the radioactive isotope is concentrated in the synovial membrane from the injected colloid suspension, where it exerts its activity. However, despite numerous reports confirming its safety and efficacy, this procedure is not widely practised. In the Singleton Hospital NHS Trust, yttrium(90) (Y(90)) RS has been practised since 1990 for refractory synovitis. In this study, we analyse the results of therapy and complications in 38 joints so treated. Doses of 10 mCi were used in the majority of patients. Most responses were apparent by 6 months following the procedure. Altogether, 68% of the treated joints showed satisfactory response at 3 years, with 29% having all symptoms under control beyond 3 years. In three patients, there was evidence of minor pigmentation at the injection site. Two patients had extravasation of the isotope and needle track ulcers, which were recorded as major toxicity. We find Y(90) radiosynovectomy to be safe, quick, and effective in the management of patients with refractory synovitis. The efficacy of RS should be tested in randomised clinical trials involving large numbers of patients.

Adult↗

Field energetics and the estimation of pollen and nectar intake in the marsupial honey possum, Tarsipes rostratus, in heathland habitats of south-western Australia.

A method is described, based on the simultaneous turnover of both stable (18O) and radioactive isotopes (3H and 22Na), whereby the daily nectar and pollen intake of free-ranging marsupial honey possums (Tarsipes rostratus) may be estimated. The field metabolic rate is measured using doubly labelled water and nectar intake is estimated independently from the measured water and sodium fluxes. The method assumes that free-water intake is negligible (but may be accounted for if not the case), that virtually all dietary sodium is derived from nectar rather than from pollen, and that the animals are in energetic balance over the period of measurement. These assumptions have been tested and found to be robust, except during periods of heavy rain when significant intakes of free-water were recorded. Leaching experiments with pollen grains suggest that less than 10% of the sodium ingested by honey possums is derived from pollen and calculations thus assumed a 90%:10% split between nectar and pollen. Nectar intake averaged 5.9 +/- 0.6 ml.day-1 and regressing nectar intake on daily change in body mass predicts an intake of approximately 7 ml.day-1 nectar to maintain balance for a 9 g honey possum. Estimates of pollen intake averaged 660 +/- 156 mg.day-1 and a similar regression analysis of the data predicts that a daily intake of approximately 1 g pollen would be needed to maintain mass balance of honey possums. Estimated nectar and pollen intakes did not differ significantly between males and females, but nectar intake was higher in winter compared with dry periods of the year. The sugar content of nectar falls during winter, however, and the overall energy derived from nectar thus remains roughly constant. Estimates of pollen and nectar intake for individual animals were not significantly correlated, suggesting that honey possums forage selectively for these two food items.

Animals↗

A Zea mays 39-kDa thylakoid transglutaminase catalyses the modification by polyamines of light-harvesting complex II in a light-dependent way.

A transglutaminase (TGase; EC 2.3.2.13) activity, which shared many properties with the TGase activity of the Helianthus tuberosus chloroplast, was observed in the Zea mays L. chloroplast and in its fractions. This activity was found to be prevalent in thylakoids; bis-(glutamyl) spermidine and bis-(glutamyl) putrescine were the main polyamine conjugates formed. Light stimulated the endogenous thylakoid activity. Putrescine, spermidine and spermine were conjugated to the isolated light-harvesting complex of photosystem II (LHCII) with different degrees of efficiency, spermine being the polyamine most efficiently conjugated. A TGase with a light-sensitive activity was identified in the photosystem II-enriched fraction. Its partial purification on a sucrose gradient allowed the separation of a 39-kDa band, which was immunorecognised by two anti-TGase antibodies (Ab-3 and rat prostatic gland-TGase). Both a colorimetric and a radiometric assay for TGase activity, the former carried out in the presence of biotinylated cadaverine and the latter in the presence of polyamines labelled with radioactive isotopes and resulting in the isolation of glutamyl-polyamines, further confirmed that the thylakoid enzyme is indeed a calcium-dependent transglutaminase (Thyl-TGase). At variance with guinea pig liver and erythrocyte TGases, which are insensitive to light, the activity of the thylakoid transglutaminase is affected by light. Moreover, this enzyme, when tested with purified LHCII as substrate, catalysed the production of mono- and bis-glutamyl-polyamines in equal amounts, whereas the 'animal' enzymes produced mainly mono-derivatives. Herein, it is discussed whether this light sensitivity is due to the enzyme or the substrate.

Animals↗

Metabolic alteration in patients with cancer: nutritional implications.

During the past 20 years, efforts have been made to elucidate the metabolic changes observed in patients with cancer by using stable and radioactive isotopic tracers. These metabolic changes in patients with cancer may be similar to those in other stress conditions, in which glucose production and utilization, lipolysis and free fatty acid flux, and net protein catabolism are increased. Stress hormones, such as glucagon and catecholamines, and certain cytokines may be responsible for these metabolic changes. Although it has been shown that cachexia in patients with cancer signals a poor prognosis, efforts to improve the clinical outcomes with nutritional support have been disappointing. The failure of cancer patients to respond to nutritional support may be related to an alteration in the intermediate metabolism. Therefore, further research evaluating the metabolic abnormalities associated with cancer may lead to more effective nutritional therapies.

Cachexia↗

The usefulness of radiotracers to make the body biochemically transparent.

Radioactive isotopes are uniquely applicable to observe reactions or circuits of reactions at the molecular level without disturbing the system being studied. The advent of molecular imaging modalities, particularly positron emission tomography (PET), is a major breakthrough for the visualisation and quantitative assessment of cellular and molecular processes occurring in living tissues. The recent development of animal PET scanners that offers 2-mm resolution and is tailored to laboratory rodent models, has made a further great impact on in vivo biochemistry. With these live-imaging modalities at hand, radiotracer-based technologies allow to look directly at biochemical distribution and interaction processes. Tremendous progress made in radiotracer chemistry, primarily in carbon-11 and fluorine-18 radiochemistry, and in the design of imaging devices strengthens the usefulness of radiotracers in nuclear medicine and drug research and development and opens exciting opportunities for new applications, e.g., in food science.

Animals↗

Random changes of amino acid residues with expected frequency by saturated point mutagenesis.

The yeast transcriptional activator protein, Gcn4p from Saccharomyces cerevisiae binds to the specific sequence in the promoters of many amino acid biosynthetic genes for general control. A new random saturation mutagenesis method was developed to isolate Gcn4p derivatives with only one or two mutations in the DNA binding domain without using radioactive isotope. This will be used to identify the amino acids of Gcn4p involved in protein-protein interactions. Saturation mutagenesis in the DNA binding domain of Gcn4p was performed using spiked degenerate oligonucleotides containing randomized codon bases designed specifically for only one or two base changes in the mutagenized area. These oligonucleotides were synthesized to have two flanking restriction enzyme sites for cloning to the appropriate vector. The 3' ends were mutually primed after hybridization via the palindromic sequences of the restriction enzyme sites. These molecules were then converted to double stranded DNA upon treatment with DNA polymerase. Here, a library collection of 100,680 in an altered Gcn4p pool was generated by cloning a mixed-base oligonucleotide in the place of the sequence coding for the DNA binding domains. The quality of the library was examined by DNA sequencing and found to be in good agreement with the expected statistical values. Calculated mutation frequency was 66% of mutant nucleotide rate and actual sequencing data revealed 68% mutant nucleotide rates from the sequenced library. Thus, among 21 mutants, 16 had one point mutations and 5 had two point mutations. This approach appears to be an effective and general tool for creating proteins with one or two amino acid change(s) in their molecules.

Binding Sites↗

Analysis of immunoglobulin light-chain mRNA in gastric malignant lymphoma using new highly sensitive in situ hybridization method.

BACKGROUND: To analyze B-cell monoclonality, we need fresh materials for the Southern blotting method. In situ hybridization (ISH) for the detection of immunoglobulin light-chain mRNA can be used in formalin-fixed and paraffin-embedded tissues. However, it is difficult to detect low levels of mRNA copies. So we must develop new highly sensitive in situ hybridization methods to diagnose gastric malignant lymphoma.METHODS: We analyzed materials from 15 patients with gastric malignant lymphoma, using a new highly sensitive ISH method, and compared the results with those of a conventional method.RESULTS: With this new method, 13 of 15 cases were shown to have monotypic cytoplasmic signals, and monoclonality was detected in 3 of 4 cases of mucosa-associated lymphoid tissue (MALT) lymphoma.CONCLUSIONS: The accuracy of endoscopic biopsies is not satisfactory. However, the detection of immunoglobulin light chain mRNA, using the new highly sensitive ISH method with a non-radioactive isotope oligonucleotide probe, can be used to diagnose gastric malignant lymphoma even when conventional immunoglobulin light-chain mRNA or proteins alone cannot demonstrate the clonality.

Journal Article↗