Synchronous fluorescence, UV-visible spectrophotometric, and voltammetric studies of the competitive interaction of bis(1,10-phenanthroline)copper(II) complex and neutral red with DNA

Constant wavelength synchronous fluorescence spectroscopy (CW-SFS), UV-visible absorption spectroscopy, and cyclic and differential pulse voltammetry were applied to investigate the competitive interaction of DNA with the bis(1,10-phenanthroline)copper(II) complex cation ([Cu(phen)(2)](2+)) and a fluorescence probe, neutral red dye (NR), in a tris-hydrogen chloride buffer (pH 7.4). The results show that both the [Cu(phen)(2)](2+)and the NR molecules can intercalate competitively into the DNA double-helix structure. The cyclic voltammetry method showed that both anodic and cathodic currents of [Cu(phen)(2)](2+) decreased on addition of the DNA and the intercalated [Cu(phen)(2)](2+)-DNA complex formed (beta = (4.14 +/- 0.24) x 10(3)). CW-SFS measurements were facilitated by the use of the three-way resolution of the CW-SFS for NR, [Cu(phen)(2)](2+), and NR-DNA. The important constant wavelength (CW) interval, Deltalambda, was shown to vary considerably when optimized (135, 58, and 98 nm for NR, NR-DNA, and [Cu(phen)(2)](2+), respectively). This approach clearly avoided the errors that otherwise would have arisen from the common assumption that Deltalambda is constant. Furthermore, a chemometrics approach, parallel factor analysis (PARAFAC), was applied to resolve the measured three-way CW-SFS data, and the results provided simultaneously the concentration information for the three reaction components, NR, [Cu(phen)(2)](2+), and NR-DNA, for the system at each equilibrium point. The PARAFAC analysis indicated that the intercalation of the [Cu(phen)(2)](2+) molecule into the DNA proceeds by exchanging with the NR probe and can be attributed to two parallel reactions. Comprehensive information was readily obtained; the replacement of the intercalated NR commenced immediately on introduction of [Cu(phen)(2)](2+), approximately 50% of NR was replaced by [Cu(phen)(2)](2+) at a concentration of 0.45 x 10(-5) mol L(-1), and nearly all of the NR was replaced at a [Cu(phen)(2)](2+) concentration of 2.50 x 10(-5) mol L(-1). This work has the potential to improve extraction of information from the fluorescence intercalator displacement (FID) assay.     

Light absorption and scattering by cell suspensions of some cyanobacteria and microalgae

Absorption spectra of cyanobacteria (Anacystis nidulans, Anabaena variabilis, and Chlorogloeopsis fritschii), red (Cyanidium caldarum and Porphyridium cruentum), green (Dunaliella maritima and Dunaliella salina) and diatom (Thalassiosira weisflogii) alga cell suspensions are presented; the spectra were obtained by using an approach developed earlier to compensate for scattering [1, 2]. In all species, the shapes of the absorption spectra were independent of the cell concentration. For Th. weisflogii and D. maritima, the analysis of selective and nonselective scattering was carried out. The effect of mechanical cell disruption on optical properties (absorption, scattering, and “package” effect) on D. maritima was studied. The character and dynamics of optical changes in D. salina under the influence of sodium chloride were followed.     

Spectroscopic determination of succinylcholine in dosage forms using eosin Y

Two simple and sensitive analytical assay methods using spectrophotometry and spectrofluorimetry techniques were developed for the estimation of succinylcholine chloride (SUC) in pharmaceutical preparations. The suggested methods are based on the formation of an ion pair complex formed between the drug and eosin Y spectrophotometrically (Method I), or the suppressive effect of succinylcholine on the native fluorescence property of eosin Y (Method II). The spectrophotometric method (Method I) involves measuring the absorbance of the complex between succinylcholine and eosin Y at 550 nm in Britton Robinson buffer of pH 3. However, the spectrofluorimetric method (Method II) involves measuring the quenching effect of the studied drug on the native fluorescence property of eosin Y at the same pH at 550 nm after excitation at 480 nm. The absorbance versus concentration of the drug is rectilinear over the range of 0.5 to 15 μg/ml. The formation constant was 3.5 × 10⁴ and the Gibb's free energy change was ?2.5 × 10⁴ J/mol. In Method II, the relative fluorescence intensity was directly proportional to SUC concentration over the range of 0.05 to 1 μg/ml. The proposed methods allowed a successful application to the estimation of succinylcholine ampoules. An explanation of the reaction pathway was postulated.     

合欢皮与山合欢皮的理化性质的比较研究

采用紫外分光光度法,薄层色谱法,溶血试验,泡沫反应等,对合欢皮与山合欢皮的理化性质进行了比较研究。结果表明:可以将山合欢皮作合欢皮使用。     

Proteomics shows Hsp70 does not bind peptide sequences indiscriminately in vivo

Heat shock protein 70 (Hsp70) binds peptide and has several functions that include protein folding, protein trafficking, and involvement with immune function. However, endogenous Hsp70-binding peptides had not previously been identified. Therefore, we eluted and identified several hundred endogenously bound peptides from Hsp70 using liquid chromatography ion trap mass spectrophotometry (LC-ITMS). Our work shows that the peptides are capable of binding Hsp70 as previously described. They are generally 8-26 amino acids in length and correspond to specific regions of many proteins. Through computationally assisted analysis of peptides eluted from Hsp70 we determined variable amino acid sequences, including a 5 amino acid core sequence that Hsp70 favorably binds. We also developed a computer algorithm that predicts Hsp70 binding within proteins. This work helps to define what peptides are bound by Hsp70 in vivo and suggests that Hsp70 facilitates peptide selection by aiding a funneling mechanism that is flexible but allows only a limited number of peptides to be processed.     

Noninvasive methods to determine the critical micelle concentration of some bile acid salts

In this work the critical micelle concentrations (cmc) of four bile salts, sodium cholate, sodium glycocholate, sodium deoxycholate, and sodium glycodeoxycholate, are determined and presented. Three independent noninvasive methodologies (potentiometry, derivative spectrophotometry, and light scattering) were used for cmc determination, at 25 degrees C with ionic strength adjusted to 0.10 M with NaCl. Spectrophotometric and potentiometric studies of some bile salts were also executed at various ionic strength values, thus allowing the influence of the ionic strength on the cmc value of the bile salt to be assessed. A critical comparison of the cmc values obtained with data collected from the literature is presented. Furthermore, this work makes an evaluation of the conceptual bases of different methodologies commonly used for cmc determination, since variations in the results obtained can be related mainly to different intrinsic features of the methods used (such as sensitivity or the need to include tracers or probes) or to the operational cmc definition applied. The undoubted definition of the experimental bile salt concentration that corresponds to cmc (operational cmc) is essential since in the case of these amphiphiles the formation of micelles is not as abrupt as in the case of ordinary association colloids. The biphasic nature of their aggregation leads to a "round-shaped" variation of the experimental parameters under analysis, which makes difficult the evaluation of the cmc values and can be responsible for the different results obtained.     

Measurement of zinc, copper, manganese, and iron concentrations in hair of pituitary dwarfism patients using flameless atomic absorption spectrophotometry

Pituitary dwarfism (hGHD) is known to be associated with trace element deficiency, which causes improper functioning of the involved endocrine system. Previously, we reported on the head hair concentrations of zinc, copper, manganese, and iron from a total of 418 normal subjects (154 male and 264 female). In this report, we analyzed the head hair concentrations of the same four trace metals of 103 hGHD children (60 male and 43 female) under treatment with human growth hormone (hGH). These subjects ranged in age from 5 to 18 yr. The results were compared with 338 agematched normal subjects (120 male and 218 female). Both male and female hGHD showed approx 1.7 times higher zinc concentrations than normal subjects. Cheruvanky et al. reported a similar trend but with a slightly lower difference between hGHD and normal subjects. The average copper content in the hair of both male and female subjects also showed higher values for the hGHD than for the normal subjects, a trend similar to the values reported by Teraoka et al. In the case of manganese, the concentrations in hair of the hGHD were approx 50% of the values in the normal subjects. Head hair concentrations of iron in the hGHD were commensurate with the normal subjects. Because the content of trace elements in hair varies with the age of subjects, as a control, we investigated the head hair concentration of zinc from 20 healthy girls ranging in age from 10 to 18 yr. The average zinc concentration decreased from 10 to 12 yr, but no clear relation to age was observed from 13 yr and older. These trends were similar to our previous report. The zinc concentration in hair and body weight gain over a year was negatively correlated. The age variation in the content of zinc, copper, manganese, and iron in hair was measured comparing hGHD with the normal subjects in various ages. Concerning the zinc-level variation of hGHD and normal subjects, there were conspicuous differences between hGHD and normal subjects. For copper, the variations in concentration with age were similar to zinc. Regarding the age variations for manganese, hGHD had lower concentrations in hair compared to the normal subjects throughout adolescence (11–18 yr). We have studied the effects between the hair and these trace element concentrations in hGHD before and after hGH administration. These results suggest that hGH affects the metabolism of these trace elements.     

Hydrolysis study of the bifunctional antitumour compound RAPTA-C, [Ru(eta6-p-cymene)Cl2(pta)

The hydrolysis of [Ru(η⁶-p-cymene)Cl₂(PTA)] (PTA = 1,3,5-triaza-7-phosphatricyclo-[3.3.1.1]decanephosphine; RAPTA-C) was studied using UV-visible (UV-vis) spectrophotometry and NMR spectroscopy. In analogy to in silico studies, [Ru(η⁶-p-cymene)Cl(H₂O)(PTA)]⁺ was found to be the most abundant hydrolysis product, although the dihydrolysed species [Ru(η⁶-p-cymene)(OH)(H₂O)(PTA)]⁺ and the dichloro compound are present. Rate constants for the different aquation and anation steps and the equilibrium constants were determined. Hydrolysis is suppressed at high chloride concentrations. These results have important implications on the mode of action of the RAPTA drug candidates.     

Determination of Mn(II) and Co(II) with Arsenazo III

Complex formation between Arsenazo III and Mn²⁺ and Co²⁺ at equilibrium has been investigated at pH 7.2, and the stoichiometry and stability of the complexes have been determined. The data indicate that Arsenazo III is suitable for determination of Mn²⁺ and Co²⁺ on the micromolar scale. The dissociation constants of the phosphate complexes of Mn²⁺ and Co²⁺ at pH 7.2 were estimated with Arsenazo III as 3.6 and 10 mM, respectively.     

Metal contamination in sediments and biota of the Bay of Quinte,Lake Ontario,Canada

The Bay of Quinte receives drainage from several large river systems, including the Moira River which carried sediment from mines into the Bay from the 1880s to the 1960s. We are investigating possible metal contamination of submerged weed beds and marsh biota which may contribute to the low diversity and biomass of macrophyte beds and Typha marshes in the Bay. In 1987, sediment, macrophytes and snails were sampled in wetlands close to the Moira River and at Hay Bay (part of the Bay of Quinte presumably unaffected by mine effluents) located 20 km from the Moira. Some element concentrations in sediment and biota were determined by neutron activation analysis (NAA) including Al, As, Br, Ca, Co, Cl, Cr, Cs, Fe, Hf, K, La, Na, Mg, Sb, Sc, Rb, Ta, Th, Ti, U, V and Zn. Other elements were analysed by acid dissolution and atomic absorption spectrophotometry (AAS) including Ag, As, Cu, Hg, Ni, Pb, and Zn. Levels of As in sediments and plants were higher close to the Moira River, whereas Cu and Ni showed the opposite pattern in sediments. The usefulness of species as bioassays differed: Stagnicola elodes Say accumulated significantly higher levels of Cu (35 vs 18 ppm) and V (l. l vs 0.5 ppm) than Planorbella trivolvis Say collected from the same sites. The macrophyte, Myriophyllum spicatum L. acted as an accumulator of Pb (up to 9.6 ppm), whereas Pb in Vallisneria americana Michx. at the same sites was undetectable.