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[Purification of cadmium ion binding metallothionein-3 by proteinase digestion on affinity chromatographic column].

The gene encoding human metallothionein-3 (hMT-3) was synthesized and inserted into the poly-cloning sites of fusion expression vector pALEX, and fused downstream to its glutathione S-transferase (GST) fusion partner. Fusion protein GST-Cd2+-hMT-3 was expressed after isopropyl-beta-D-thiogalactopyranoside (IPTG) induction and addition of 0.1 mmol/L CdSO4 into the culture medium, and mainly existed in cellular soluble fraction as revealed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Recombinant MT was purified by two purification procedures: "proteinase digestion after purification" method, e. g. by elution of GST-Cd2+-hMT-3 from GST affinity chromatography first, then proteinase digestion and GST affinity chromatographic purification again subsequently; and "proteinase digestion in situ" method, e. g. digestion of GST-Cd2+-hMT-3 directly on the column while its binding to the GST affinity chromatographic resin and collection of Cd2+-MT directly from the flow through fraction after the digestion. It was confirmed that the later procedure exhibited more effective and more convenient by avoiding the conventional elution, dialysis and lyophilization processes and increasing the purity, recovery or yield of the final product. After further purification by a Superdex 75 HR 10/30 column, finally 6-7 mg of Cd2+-hMT-3 was obtained from 3 L of flask culture with the recovery of about 1.8%. SDS-PAGE, amino acid composition and inductively coupled plasma atomic emission spectrometer (ICP-AES) analysis showed that the relative molecular mass of Cd2+-hMT-3 is about 7 000, with a purity above 90%. Its amino acid composition is consistent with the expected value of natural hMT-3, particularly no aromatic amino acid and histidine, and the atomic ratio of 21: (7.5 +/- 0.1) for S: Cd, is also consistent with the theoretical value of 21: 7.

Cadmium↗

[Synthesis and spectral studies of functionalized L-Cys-CdS nanoparticles as fluorescence probes].

In this work, nano-CdS was successfully prepared. The nano-CdS was also modified with L-cysteine. Absorption and fluorescence spectra of CdS nanoparticles for different pH values, reaction times and different Cys/Cd2+/S ratios were investigated. Meanwhile a fluorescence enhancing effect was observed between trace zinc ions and the functionalized L-Cys-CdS nanoparticles. The response is linearly proportional to the concentration of zinc ions from 1.0 to 15 micromol x L(-1). The functionalized nanoparticles are hopeful of use as fluorescence probes in detecting trace elements in biological samples.

Cadmium Compounds↗

Application of a continuous ventricular function monitor with miniature cadmium telluride detector to patients with coronary artery bypass grafting.

A continuous ventricular function monitor with a miniature cadmium telluride detector was evaluated and applied to patients with coronary bypass surgery (CABG). Ejection fraction (EF) at rest and change in EF from rest to exercise and postexercise (Y) measured with the device correlated with that of the gamma camera (X) (Y = 0.86x + 6.8(%), r = 0.87, n = 110, p less than 0.001, and Y = 0.96x + 0.4(%), r = 0.90, n = 37, p less than 0.001 respectively). Left ventricular function during and after supine ergometer exercise was monitored in 54 patients before and after CABG. The EF change from baseline to peak exercise improved from -5.9% +/- 8.9% before CABG to 7.2% +/- 7.9% after CABG (p less than 0.001). In all patients but two, a rapid EF increase just after exercise over baseline EF was observed. This EF "overshoot" during recovery increased from 11.5% +/- 6.5% to 16.4% +/- 6.0% (p less than 0.001) after CABG. The time from the cessation of exercise to EF overshoot decreased from 153 +/- 80 sec to 76 +/- 49 sec (p less than 0.001) after CABG. The continuous ventricular function monitor with a miniature cadmium telluride detector is able to measure EF reliably. Following successful aortocoronary bypass, EF response during exercise improved and the EF overshoot in the recovery phase became faster and higher.

Cadmium↗

133Xenon absorption into rubber-protected portable cadmium telluride (CdTe(Cl)) detectors invalidating the 133Xenon washout method for measurement of cutaneous and subcutaneous blood flow rates in man.

The importance of 133Xenon absorption into rubber detector caps during cutaneous and subcutaneous blood flow measurement was investigated in 46 experiments involving 38 persons. 133Xenon was administered atraumatically. Cutaneous and subcutaneous washout rates were registered by portable Cadmium Telluride detectors without rubber caps, with rubber caps, and with rubber caps with Mylar membranes interposed between the rubber and the tissue investigated. No difference in rate constants obtained by means of various detector types was detected. The accumulation of 133Xenon in the rubber caps was found to take place within the first few minutes after the detectors had been brought into position. The 133Xenon then diffused back into the tissue exhibiting a great variation regarding rate constants. The 133Xenon diffused form rubber into air and perfused tissue tracing a monoexponential course; and again the rate constants would vary considerably. No correlation was found between elimination rates obtained with detectors with and without 133Xenon polluted caps, and no way of correcting for the 133Xenon content in the rubber caps was found. Relative changes in rate constants could still be recognized, but absolute values were not obtainable.

Absorption↗

[The formation of an adaptive genetic system in an inbred strain of Drosophila melanogaster selected for high embryonic mortality].

In prolonged genetic experiment two contrast inbred strains, selected for the differences in embryonic mortality have been produced (HEM and LEM). In HEM strain the balanced lethal system in 55.0 region (near B1) of chromosome 2 has been formed. This system provides a high level of average fitness in the strain, despite of the high level of embryonic mortality.

Adaptation, Physiological↗

[Cadmium-induced changes in the activity of the dopaminergic and purinergic systems and in ascorbic acid catabolism in the rat striatum].

Levels of dopamine (DA), 3-4-dihydroxyphenylacetic acid (DOPAC), ascorbic acid (AA), dehydroascorbic acid (DHAA), uric acid (UA) and adenosine (ADO), were determined by HPLC in the striatum of male Wistar rats treated with repeated injections of cadmium (as sulfate) 3 mg/kg/day s.c. for 10 consecutive days. Cadmium treatment significantly reduced DOPAC levels with consequent decrease of the DOPAC/DA ratio; AA levels were significantly reduced, while DHAA levels were significantly increased, with consequent increase of DHAA/AA ratio: levels of UA and ADO were both significantly increased. It is concluded that cadmium, given systemically, reduces dopaminergic system activity in the rat striatum; such impairment occurs together with an increase of AA oxidation and of markers of purinergic system activity. The inhibition of striatal dopaminergic activity could be considered the neurochemical basis of behavioral changes induced by cadmium, while the increase of AA oxidation and of purinergic system activity could be considered an antitoxic metabolic response.

Adenosine↗

[Basic evaluation of an ambulatory ventricular function monitor with a a cadmium telluride detector].

The reliability of a new ambulatory ventricular function monitor (VEST) with cadmium telluride (Cd-Te) detector was validated. No counting loss was observed under 20 kcps. In phantom study, more than 200 ml with 6 cm distance from the detector and more than 300 ml with 6-12 cm distance, underestimation of volume was observed. Ejection fraction (EF) calculated in phantom study showed close value to the true EF in 50-400 ml of end-diastolic volume. In 10 patients, 1-2 cm detector shift toward right upper, upper and left showed underestimation of EF. Good correlation of gamma camera EF (X) and VEST EF (Y) was obtained (Y = 0.96X + 3.6(%), n = 37, r = 0.93, p less than 0.001). EF change from rest values, obtained from each comparable exercise and post exercise stage by gamma camera (X) and VEST (Y), showed good correlation (Y = 0.95X + 0.8, n = 20, r = 0.85, p less than 0.001). In conclusion Cd-Te VEST was considered as a reliable EF measurement technique similar to gamma camera.

Cadmium↗

Enhanced production of hepatitis B virus surface antigen in mouse C127 cell on a bovine papillomavirus-metallothionein vector.

We have constructed a recombinant plasmid pCPS12 containing the hepatitis B viral surface antigen (HBsAg) gene linked to the mouse metallothionein promoter on a BPV-pML2 vector. Two stable clones S12-8 and S12-2, obtained by transfection of the mouse C127 cells with pCPS12 propagated in dam+ dcm+ and dam- dcm- Escherichia coli respectively, exhibited different types of response to 5-azacytidine (5-aza-CR) and cadmium (Cd) induction. In S12-8, the productivity of HBsAg was enhanced by 5-aza-CR or 5-aza-CR plus Cd, but not by Cd alone. In S12-2, the expression of HBsAg was not affected by 5-aza-CR but was induced by Cd in the presence or absence of 5-aza-CR. This suggests that methylation may be important in controlling the HBsAg expression and the inducibility of Cd in the transfectants.

Animals↗

Glutathione-coated cadmium-sulfide crystallites in Candida glabrata.

Cadmium-sulfide crystallites form in the yeast Candida glabrata cultured in the presence of cadmium salts. The particles function to sequester and detoxify intracellular cadmium ions. The crystallites are peptide-coated, but the coating peptide varies with the nutrient conditions of the growth medium. When cultured in rich nutrient broth the yeast forms intracellular CdS particles coated with a mixture of glutathione and the gamma-glutamylcysteine dipeptide. In contrast, cultures in synthetic minimal medium yield particles coated with polymerized gamma EC peptides of general structure (gamma-Glu-Cys)n-Gly. Glutathione/gamma-glutamylcysteine particles exhibit properties analogous to quantum, semiconductor-type crystallites. The optical properties are dependent on particle size, and irradiation results in photoluminescence and photoreduction not observed in bulk CdS mineral. Aerobic irradiation leads to particle decomposition presumably via oxidation of the sulfide ions within the crystallite.

Cadmium↗

[Cloning of the gene of Saccharomyces cerevisiae yeasts that determines resistance to the toxic action of cadmium ions].

A gene conferring resistance to cadmium in Saccharomyces cerevisiae was isolated from a yeast gene library created on the basis of the pL3 vector. The phenotype of resistance is only expressed in the yeast cells with cloned DNA inserted into a multicopy plasmid. Integration of the plasmid into chromosome or introduction of the centromeric region into the plasmid decreases the level of cadmium resistance. The cloned Sau3A I fragment of the yeast chromosome is 3.5 kbp in size. Restriction analysis and subcloning experiments showed the gene to be located within 1.6 kbp of the XhoI-Sau3A I fragment of DNA. Instability was observed in the vicinity of the XhoI-Sau3A I fragment of the yeast DNA in Escherichia coli.

Base Sequence↗

Sensitivity enhancement of ninhydrin-treated latent fingerprints by enzymes and metal salts.

Lyophilized Sigma Type III trypsin has been applied to latent prints two weeks to two months old. This trypsin preparation eliminates the background problems that had been encountered with old prints in a previous study. Zinc chloride treatment of latent prints previously exposed to ninhydrin enhances their detectability upon laser examination. However, it has been reported that the zinc chloride reaction occasionally fails to occur. Accordingly, we have investigated the optimization of this reaction. We find that high humidity and elevated temperature, particularly the former, are needed. Cadmium nitrate, although it produces weaker fluorescence than zinc chloride, may at times be useful. Reaction conditions are much the same as those for zinc chloride.

Cadmium↗

Experimental teratological studies with the mouse CNS mutations cranioschisis and delayed splotch.

Teratological experiments were made with a recessive mouse gene (cranioschisis) causing exencephaly and a semidominant gene (delayed splotch) causing spina bifida. In studies with the cranioschisis gene administration of warfarin and thyroxine resulted in frequencies of exencephaly significantly below that expected of a recessive trait, perhaps indicating selective elimination of abnormal conceptuses. Studies with the delayed splotch gene tested the hypothesis that offspring with a hereditary defect of neural-tube closure have other, unexpressed CNS defects, which may be elicited by teratological impulses. This proposition was decisively upheld by administering 5-bromo-2'-deoxyuridine, cadmium sulfate and retinoic acid, as these treatments all caused significantly greater frequencies of induced exencephaly in offspring with spina bifida than in their genetically normal littermates.

Animals↗