Optimization of Total Thyroxine Radioimmunoassay in Unextracted Serum by Dextran Charcoal

Specific method for accurate radioimmunoassay of free and total thyroxine in un-extracted serum has been developed. The method is used for estimation of either free thyroxine (T₄) or total T₄ by inhibition of the binding sites of T₄ to protein using TCA/NaOH mixture. The antibodies used in this study were obtained from Mallinkrodt. The separation technique used in our modification is that of dextran charcoal, this is to absorb the free fractions on charcoal.

The kinetics of antibody antigen combination, the parameters of the experiment like; charcoal concentration, antibody dilution, temp., time of incubation and radioactivity concentration were, studied to optimize the standard curve and the sensitivity of T₄RIA.

Serum samples of different thyroid cases were investigated for determination of free and total T₄ concentration to get the levels for each group of which 50 normals, 57 hyperthyroidism and 21 hypothyroidism in comparison with the classical tests of thyroid functions. Also sensitivity and variance coefficient has been carried out in comparison with other T₄RIA techniques.     

Studies on Trace Labelling of Bromsulphalein (BSP) with Radioiodine

Two methods have been studied for high specific activity trace iodination of Bromsulphalein (BSP) with radioiodine (¹³¹I). In the first method, BSP in aqueous solution is reacted with iodine and an oxidising agent – hydrogen peroxide or chloramine T. In the second method, iodination is carried out by electrolytically liberated iodine. The electrolytic method gives labelled BSP free from any labelled degradation products. The stability of ¹³¹I-labelled BSP in aqueous solution has been studied. The product has been found to remain more stable in aqueous solutions at a slightly acid pH (4…6.5) than in dilute alkaline solutions (pH 7…9.5).     

Studies on the Purity of 51Cr Labelled Human Serum Albumin

Different methods reported for the purification of Chromium-51 labelled human serum albumin have been studied in detail. It has been found that purification by ion exchange or dialysis does not remove some of the impurities such as complexed chromium and hydrated chromic oxide. Purification over DEAE cellulose or Sephadex (A-50) gives a product which is free from these impurities. It has been confirmed that chromium labelling leads to the formation of labelled protein aggregates and the specific activity of the labelled aggregates is greater than that of the non-aggregated fraction. These aggregates can be separated from the labelled monomer by column chromatography of Gel filtration.     

Doping and radiation damage profiles of P+ ions implanted in silicon along the [110] axis

Several doses of 200 KeV phosphorus ions have been implanted under channeling conditions along the [110] direction in silicon.

Range distribution has been determined for the three implant doses 10¹³, 10¹⁴, 10¹⁵ P⁺/cm² both with the electrical measurements and the neutron activation techniques.

The radiation damage distribution has been determined both with 290 KeV proton back-scattering analysis and with transmission electron microscopy (TEM) observations.

Good agreement has been found between electrical and neutron activation profiles in the samples where 100% of the implanted dose had been electrically activated by means of annealing.

Carrier concentration profiles, from samples implanted with 10¹⁵ P⁺/cm², determined after two different annealing temperatures (500°C and 700°C) have bcen compared with the radiation damage distribution and a correlation between damage and phosphorus electrical activation process seems to be possible.

Maximum damage peak, as determined by back-scattering analysis, shifts from ～0.4 μ depth in the lower dose(5 × 10¹⁴ P⁺/cm²), to ～0.22 pm depth in the higher implanted dose (4 × 10¹⁵ P⁺/cm²). Damage distribution of phosphorus ions random implanted in the same experimental conditions shows 3 peak at ～0.2 μn depth.

In accordance with the back-scattering analysis, T.E.M. observations on 10¹⁵P⁺/cm² implanted samples show the presence of amorphous regions at depth between 0.25 and 0.5 μm from the surface. In the most damaged layer ～0.3μm in depth, a surface density of ～10¹²/cm² amorphous regions 25-50 A diameter was observed.     

13C/12C-Untersuchungen an Tumorzellen

Abstract

The metabolism of tumor-cells differs in many ways from normal (healthy) cells. One of the major differences is the high glycolytic activity in tumor-cells with the subsequent formation of lactate from glucose, even in the presence of oxygen. The question whether this high rate of glycolysis has any effect on the ¹³C/¹²C-relation of the cells is examined in experiments with a tumor-cell line (HT29) and in specimens of human breast cancer.

The HT29 cells show a clear decrease in ¹³C content compared to their culture medium (Δδ = 3.28‰).

Tissue from human breast cancer has more ¹³C than normal breast tissue taken from the same patient ((Δδ = 2.74‰). But the content of fat is much higher in the normal tissue and its δ-value is negatively correlated with its fat content. It is concluded that the difference between normal and tumor tissue is due to the heterogeneous composition of the normal tissue samples.     

Infrared Study of Anthraquinone-Anion-Radical and its 180 Labelled Isomer

Abstract

Infrared spectra of anthraquinone and anthraquinone-¹⁸0-anion-radicals have been investigated. Anion-radicals have been obtained by electrochemical reduction of anthraquinone and anthraquinone-¹⁸0 in dimethylsulf-oxide-d₆.

Band assignment has been performed using the group vibrational concept and isotopic shift data. The assignment of IR active C[dbnd]O stretching band and the other bands in 1150–1800 cm^?1 region of anthraquinone anion-radical (anthrasemiquinone) have been discussed and compared with those of some other ¹⁸0 labelled anion-radicals. A decrease of 182 cm^?1 has been found forafter transformation of the neutral quinone into anion-radical.     

Incorporation of Radioiodine in Thyroid Hormones for the Determination of T3/T4 Ratio Using Paper Chromatography

A precise and accurate method is being developed for the determination of T₃/T₄ ratio. This method depends on using serum of different thyroid cases for the assesment of T₃/T₄ ratio. It has been found that the best temperature of storage is 0.0 °C to 4 °C before extracting the iodinated tyrosines and thyronines.

Iodination with ¹²⁵I was done according to our previous technique to render the separated thyronine radioactive. Scanning method helped for the determination of T₃/T₄ ratio. The results were confirmed by sensitivity and recovery test and statistically analyzed. 100 cases were investigated comprising hypo-, eu-, and hyperthyroid cases.     

On the Preparation and Distribution of two S35-Labelled Hypoglycemic Agents

Two S³⁵-labelled hypoglycemic agents were prepared by the interaction of labelled sulphonamide and urea derivatives. The R values of the intermediates as well as the products wer determined and compared with tolbutamide. The purity of the products was checked by melting point determination, infrared spectrum, and radioactivity investigations. Localization of the separated products on the chromatograms was done by colour reaction, radioscaning, and radioautography. Distribution of the labelled drung in different tissues of rats was also carried out to prove the site of action of the drung and to show its accumulation. Counting of these tissues has been carried out by means of a liquid scintillation counter type Nuclear Chicago.     

Towards an Inhalative 13C Breath Test Method

Abstract

Customary ¹³CO₂ breath tests—and also ¹⁵N urine tests—always start with an oral administration of a test substrate. The test person swallows a stable isotope labelled diagnostic agent. This technique has been used to study several pathophysiological changes in gastrointestinal organs. However, to study pathophysiological changes of the bronchial and lung epithelium, the inhalative administration of a stable isotope labelled agent appeared more suitable to us. [1-¹³C]Hexadecanol and [1-¹³C]glucose were chosen. Inhaled [1-¹³C]hexadecanol did not yield ¹³CO₂ in the exhaled air, but [1-¹³C]glucose did. To study the practicability of the [1-¹³C]glucose method and the reproducibility of the results, 18 inhalation tests were performed with healthy subjects. In 6 self-tests, the optimum inhalative dose of [¹³C]glucose was determined to be 205 mg. Using the APS aerosol provocation system with the nebulizer ‘Medic Aid’ (Erich Jaeger Würzburg), a 25% aqueous solution was inhaled. Then, breath samples were collected at 15 min. intervals and analysed for ¹³CO₂. 75–120min after the end of inhalation a well-reproducible maximum δ¹³C value of 6‰ over baseline (DOB) was detected for 12 healthy probands.

Speculating that the pulmonary resorption of the [¹³C]glucose is the rate-limiting step of elimination, decompensations in the epithelium ought to be reflected in changed [1-¹³C]glucose resorption rates and changed ¹³CO₂ output.

Therefore, we speculate that the inhalation of suitable ¹³C-labelled substrates will pave the way for a new group of ¹³CO₂ breath tests aiding investigations of specific pathophysiological changes in the pulmonary tract, such as inflammations of certain sections and decompensations of cell functions.     

Analysis of 13C-mixed Triacylglycerol in Stool by Bulk (Ea-IRMS) and Compound Specific (GC/MS) Methods

Abstract

This paper was presented in poster form at the 17th International Congress of Nutrition, August 27-31, Vienna. Austria (Annals of Nutrition & Metabolism 2001; 45(Suppl.1):349). Some of the data were also presented in poster form at the British Society of Gastroenterology Meeting, March 18-21, Glasgow, UK (Gut 2001; 48(Suppl.1):A91).

The ¹³C-mixed triacylglycerol (MTG) breath test is used to measure intraluminal fat digestion. In normal digestion. 20–40% of the ingested ¹³C label is recovered in breath CO₂. We aimed to identify the proportions of ingested label excreted in stool, as well as breath following ingestion of ¹³C-MTG by children with impaired exocrine pancreatic function and healthy controls. ¹³C enrichment of breath samples was measured by continuous flow isotope ratio mass spectrometry (IRMS) and cumulative percent dose recovered (cPDR) in 10 h was calculated. Total ¹³C of a faecal fat extract from each stool was measured by elemental analyser-IRMS, and ¹³C enrichment and concentration of the TBDMS derivative of octanoic acid was measured by GC/MS after hydrolysis of the fat extract. Stool 5-day cPDR was calculated. Mean breath cPDR was 35%. Mean cPDR in stool by combustion-IRMS and GC/ MS, respectively, was 0.8% and 1.0%. Therefore, the remaining 64% of the ¹³C label must remain in the body and variability in breath cPDR is due to postabsorptive rather than predigestive factors.     

A Possible Relationship Between Serum Satellite DNA and Cellular Antioxidative Mechanism

The presence of circulating free DNA in the sera of cancer patients has been reported by different authors. Moreover, a hypothetical relationship between the DNA level and different stages of the disease has also been suggested. In this work, the free serum DNA level in various cancer patients was measured and consistent results with the reported data were found. (1,4) To assess the possible mechanism for the release of this fraction and to explain the high levels observed in cancer patients levels of the bivalent intracellularcation, magnesium were determined. Further to relate the process to the antioxidative mechanism of the cell membrane also selenium levels were measured simultaneously. The serum free DNA and the magnesium concentration of the patients before treatment were significally higher and the selenium levels lower than the control group. After appropriate the rapy the magnesium levels were found to decline while the high free DNA and low selenium values still persisted. These findings imply that the membrane stability and permeability is impaired due both to the neoplastic changes and later to the effect of chemotherapeutic agents with the result that intracellular material is released into the circulation.     

Measurement of uranium and its isotopes at trace levels in environmental samples using mass spectrometry

Actinides have widely entered the environment as a result of nuclear accidents and atmospheric weapon testing. These radionuclides, especially uranium, are outstanding radioactive pollutants, due to their high radiotoxicity and long half-lives. In addition to this, since depleted uranium (DU) has been used in the Balkan conflict in 1999, there has been a concern about the possible consequences of its use for the people and environment. Therefore, accurate, precise and simple determination methods are necessary in order to evaluate the human dose and the concentration and effects of these nuclides in the environment. The principal isotopes of uranium e.g. ²³⁵U and ²³⁸U are of primordial origin and ²³⁴U present in radioactive equilibrium with ²³⁸U. ²³⁶U occurs in nature at ultra trace concentrations with a ²³⁶U: ²³⁸U atom ratio of 10⁻¹⁴. Concentrations of uranium in soil samples were determined using inductively coupled plasma mass spectrometry (ICP-MS) and isotope ratios of uranium were measured using a thermal ionisation mass spectrometer. Radioactive dis-equilibrium of ²³⁴/²³⁸U, depletion of ²³⁵/²³⁸U and significant evidence of ²³⁶U/²³⁸U were noticed in soil samples.      

Radiation effects on luminescent and structural properties of YPO4: Pr3+ nanophosphors

ABSTRACT

YPO₄ phosphors doped with trivalent ion Pr³⁺ were prepared by sol–gel method and treated with different doses of gamma radiation, from 0.25 MGy to 4 MGy. Effects of radiation on morphology, structure and luminescent properties were analyzed. Also, the influence of radiation on the change in the color of the samples was examined. The color efficiency of powders was evaluated by colorimetric analysis (CIE and L * a * b system). It has been observed that powders change color under the influence of radiation, i.e. they pass from white to pinkish red. Also, it has been determined that the radiation affects morphology change, as the particle size increases with increasing of the radiation dose. With the increase in the radiation dose, the emission intensity of samples decreases. The structure remains almost unchanged after irradiation, and the intensity constantly decreases with increasing of dose.     

Investigation of the Oxidation of Serum Albumin by Iodine

In studies of metabolism using serum albumin labelled with iodine (I¹³¹ and I¹²⁵) the question of the biological equivalence of the labelled material is of decisive importance. Changes of the chemical and biological characteristic of various proteins after iodination have been found by a number of workers [1–4], especially with reference to chromatographic behaviour, rate of metabolism in the organism and content of thiol. Oxidation of sulphhydryl groups can lead to interchain disulphide bridge-linkage and the formation of molecule aggregates.     

Efficiency of Refining of Acetic Acid from Inorganic Impurities by Normal Freezing Method by Radioisotopic Technique

Investigations on efficiency of refining of acetic acid by normal freezing method with radiotracer technique and neutron activation are presented. Equilibrium distribution coefficient for Zn²⁺ in CH₃COOH was determined; the same coefficient was also determined for Cl^? but with lower degree of approximation and a “generalized” distribution coefficient for a set of ions was estimated by comparison of radioactivity of original and refined samples of acetic acid after neutron activation in nuclear reactor.     

Part 1: An Appraisal of New Radioisotope Techniques in the Study of Iodine Metabolism

The use of different radioiodines and technetium for the evaluation of the thyroid functions has been discussed. Preparation oe labelled iodotyrosines and labelled throxine is mentioned for the convey of different radiochemical tests and for the use of reference standards, necessary for the identification of separated amino acids. Paper chromatography and paper electrophoresis techniques are used for the identification and determination of iodinated amino acids or homones, either in the free state or bound to the proteins. Tc^99m is employed to substitute radioiodine in the study of uptake measurements of the thyroid gland and to evaluate the excretion of the isotope in urine. Thiourea-S³⁵ is employed, for the first time to human, to study the metabolism of the substance in different thyroid disorders.     

Spectra of 2, 5- and 3, 4-Dichlorobromobenzene Molecules in Ultraviolet

Vibrational analyses have been presented for the absorption bands of 2, 5- and 3, 4-dichlorobromobenzene molecules photographed in the regions 2925 - 2690 A and 2900 - 2580 A respectively. The following constants have been determined:

The 0.0 band shifts have been explaine on the basis of simple additivity rule.     

The Fate of [15N]Ammonium and [15N]Nitrate in the Soil of a 140-Year-Old Spruce Stand (Picea Abies) in the Fichtelgebirge (NE-Bavaria)

Abstract

A ¹⁵N tracer-experiment was carried out in a 140-year-old spruce stand (Picea abies (L.) Karst.) in the Fichtelgebirge (NE-Bavaria, Germany). Highly enriched (98 at%) [¹⁵N]ammonium and [¹⁵N]nitrate were applied as tracers by simulation of a deposition of 41.3 mol N ha^?1 with 11 water m^?2. To examine seasonal variations of uptake by spruce and understorey vegetation, different plots were labelled in spring, summer and autumn 1994.

One aim of the present study was to perfect a method of preparation of soil extracts for isotope ratio mass spectrometry (IRMS) measurements. Ammonium and nitrate from soil extracts were prepared for IRMS measurements by steam distillation and subsequent freeze drying. Additionally, tracer distribution and transformations in the soil nitrogen pools were examined. Ammonium, nitrate and total nitrogen were examined in the organic layer and the upper 10 cm of the mineral soil during 3 months after the first tracer application in spring 1994.

In July 1994, three months after tracer application, 40% of the [¹⁵N]ammonium label and 29% of the [¹⁵N]nitrate label, respectively, were recovered in the total N pool of the investigated soil horizons. In the organic layer the L/Of horizon retained most of the recovered tracers. Nitrification, immobilisation and mineralisation occurred even under the conditions of high soil acidity at the study site.     

Loss of Trace Elements From Natural Water During Storage 2. Behavior of 203HgCl2, CH3 203HgCl and 65ZnCl2 Added to Marine Water

Dilute solutions (10–50 ppb) of mercury and zinc are prepared using artificial marine water and natural marine water, labelled with ²⁰³Hg and ⁶⁵Zn.

²⁰³Hg in inorganic form is lost considerably from both solutions caused by vaporization to the atomosphere and presumably by adsorption to the surface of containers. In natural marine water some biological activities participate for the behavior of both metals but it seems to be in smaller extent than that in pond water. ⁶⁵Zn remained constant with no loss under these experimental conditions.

When CH₃ ²⁰³HgC1 is added to the solution instead of ²⁰³HgC1₂, ²⁰³Hg is scarcely lost but a smaller amount is observed to be lost by vaporization from well-test tubes. In natural marine water, however, CH₃ ²⁰³HgC1 is also lost in the similar manner as ²⁰³HgC1₂.     

Relationship of the Acid-Induced 13C Chemical-Shift Change in the α-Carbon Atom to the Stretching Frequency of the Hydroxy Group in Saturated Alcohols

A good linear relationship was found between the acid-induced ¹³C chemical shift change of the α-carbon atom and the OH stretching frequency of the hydroxy group in some saturated alcohols. The relationship is discussed with model calculations using the CNDO/2 method.

The trifluoroacetic acid-induced ¹³C chemical shift change in the α-carbon atom to the hydroxy group, Δδ¹³c, has been measured for a number, of saturated alcohols in carbon tetrachloride solution and provides information on the relative basicity and its dependence on the structural factor.¹ The free OH stretching frequency, v_OHis known to depend on the structural factor of the substituent._2–5 We reported that a linear relationship exists between the ¹⁷ O chemical shift, δ¹⁷ O, and the OH stretching frequency of the hydroxy group in saturated alchohols, R-OH.⁵ In this case, the ¹⁷O chemical shift change was reported to be governed by the ionization potential.⁶ The ionization potential of R-OH is expected to be correlated with its basicity. Regression analysis for Δδ¹³C was thus carried out for Δδ¹⁷OH reported for 14 and 19 typical alcohols, respectively. In order to explain the experimental results the oretically, we carried out CNDO/2 calculations for a series of methyl-substituted methanols.