Trace elements in sickle cell disease

Instrumental Neutron Activation Analysis (INAA) and Proton-Induced X-ray Emission (PIXE) analysis (employed as a complementary technique) have been used to determine the concentration of 11 elements in blood samples and its components erythrocytes and plasma obtained, from three groups of subjects in Nigeria viz: sickle cell anaemia (SCA) subjects, subjects with sickle cell trait and normal control subjects. The results suggest that SCA subjects have significantly higher concentrations of Na, Cl, Ca and Cu in their whole blood and erythrocytes and a higher concentration, of Cl and Cu in their plasma relative to the control subjects. Furthermore, a significantly lower concentration of K, Fe, Zn, Se, Br and Rb were found in the whole blood and erythrocytes of the SCA subjects as compared to the controls while the concentration of K and Fe in the plasma of the SCA subjects were however, found to be significantly higher than that of the control group. The study also shows that there was no significant differences between the concentration of these 11 elements in the group with sickle cell trait and the normal control group.     

Molecular aspects of sickle cell disease.

Sickle cell hemoglobin (hemoglobin S) contains valine instead of glutamic acid in position 6 of the β-chain. Few detectable conformational differences exist between hemoglobin S and normal adult hemoglobin (hemoglobin A). Following loss of oxygen, sickle cell hemoglobin (hemoglobin S) self associates to form a liquid crystal phase which distorts the erythrocyte into the sickle shape thereby resulting in the clinical symptomology associated with sickle cell anemia. This gel contains deoxyhemoglobin S monomers and polymers in equilibrium. The polymerization process is known to have a negative temperature coefficient, to be pH dependent, and to be extremely dependent on hemoglobin S concentration. The polymerization of deoxyhemoglobin S appears to be entropically driven and occurs in two kinetic phases, a delay period and a rapid polymerization process. The polymer consists of tubules containing six or eight strands of deoxyhemoglobin S tetramers which align with one another. Each strand is would around the tubule with a pitch of about 3000 Å, but the precise nature of the intermolecular hemoglobin S contacts is not known. Subsequent alignment of the tubules occurs and results in the tactoids observed in (S/S) erythrocytes. While many of the details of polymer formation and structure remain to be elucidated, several attempts to chemically alter the sickling phenomenon have been carried out. As yet, however, no satisfactory chemical treatment has been discovered.     

Dielectric spectroscopy of red blood cells in sickle cell disease

Hypoxia-induced polymerization of sickle hemoglobin and the related ion diffusion across cell membrane can lead to changes in cell dielectric properties, which can potentially serve as label-free, diagnostic biomarkers for sickle cell disease. This article presents a microfluidic-based approach with on-chip gas control for the impedance spectroscopy of suspended cells within the frequency range of 40 Hz to 110 MHz. A comprehensive bioimpedance of sickle cells under both normoxia and hypoxia is achieved rapidly (within ∼7 min) and is appropriated by small sample volumes (∼2.5 μL). Analysis of the sensing modeling is performed to obtain optimum conditions for dielectric spectroscopy of sickle cell suspensions and for extraction of single cell properties from the measured impedance spectra. The results of sickle cells show that upon hypoxia treatment, cell interior permittivity and conductivity increase, while cell membrane capacitance decreases. Moreover, the relative changes in cell dielectric parameters are found to be dependent on the sickle and fetal hemoglobin levels. In contrast, the changes in normal red blood cells between the hypoxia and normoxia states are unnoticeable. The results of sickle cells may serve as a reference to design dielectrophoresis-based cell sorting and electrodeformation testing devices that require cell dielectric characteristics as input parameters. The demonstrated method for dielectric characterization of single cells from the impedance spectroscopy of cell suspensions can be potentially applied to other cell types and under varied gas conditions.     

Synthetic studies of cystobactamids as antibiotics and bacterial imaging carriers lead to compounds with high in vivo efficacy

There is an alarming scarcity of novel chemical matter with bioactivity against multidrug-resistant Gram-negative bacterial pathogens. Cystobactamids, recently discovered natural products from myxobacteria, are an exception to this trend. Their unusual chemical structure, composed of oligomeric para-aminobenzoic acid moieties, is associated with a high antibiotic activity through the inhibition of gyrase. In this study, structural determinants of cystobactamid''s antibacterial potency were defined at five positions, which were varied using three different synthetic routes to the cystobactamid scaffold. The potency against Acinetobacter baumannii could be increased ten-fold to an MIC (minimum inhibitory concentration) of 0.06 μg mL⁻¹, and the previously identified spectrum gap of Klebsiella pneumoniae could be closed compared to the natural products (MIC of 0.5 μg mL⁻¹). Proteolytic degradation of cystobactamids by the resistance factor AlbD was prevented by an amide-triazole replacement. Conjugation of cystobactamid''s N-terminal tetrapeptide to a Bodipy moiety induced the selective localization of the fluorophore for bacterial imaging purposes. Finally, a first in vivo proof of concept was obtained in an E. coli infection mouse model, where derivative 22 led to the reduction of bacterial loads (cfu, colony-forming units) in muscle, lung and kidneys by five orders of magnitude compared to vehicle-treated mice. These findings qualify cystobactamids as highly promising lead structures against infections caused by Gram-positive and Gram-negative bacterial pathogens.

Structure–activity relationship studies of the natural product cystobactamid at four different positions led to novel imaging probes and analogs with superior antibacterial activities and in vivo efficacy.     

Isotopic fingerprint method: Assessment of the origin of rare earth compounds from the isotope ratios of lead impurities

The isotope ratios ²⁰⁷Pb:²⁰⁶Pb and ²⁰⁸Pb: ²⁰⁶Pb are measured by means of inductively coupled plasma mass spectrometry (ICP-MS) for monazite minerals and commercial rare earth compounds and are evaluated with respect to the initial composition of the minerals and the origin of the samples. The application of the isotopic fingerprint method is demonstrated.Dedicated to Professor Dr. Dieter Klockow on the occasion of his 60th birthday     

Potentiometric microdetermination of lead(II) with an ion-selective lead electrode and its application to the analysis of organic lead compounds

Summary A method for the microdetermination of lead(II) by direct potentiometric titration with 0.0025 M sodium molybdate is described. A lead-selective indicator electrode and a double-junction reference electrode are used in conjunction with an expanded-scale pH-meter to detect the end-point. If the titrant is standardized against 1.5 mg of lead(II), accurate results are obtained for 1–2 mg of lead: the absolute error does not exceed 3gmg of lead. The relative standard deviation is 0.05%. The optimum pH for the titration is 5–5.5. In water-organic solvent mixtures the potential break at the endpoint increases, but the electrode response becomes very slow. The titration of lead is also possible in sodium nitrate and/or perchlorate media, so this titration is applicable to the microdetermination of lead in organic lead compounds after wet mineralization with HNO₃ and/or HClO₄. Chloride (up to 1000 times the lead content) and bromide (100 times) do not interfere. Iodide and sulphate interfere. Five organic lead compounds have been analysed successfully: the standard deviation of the absolute error was 0.1%.

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Zusammenfassung Ein Verfahren zur Mikrobestimmung von Blei(II) durch direkte potentiometrische Titration mit 0,0025M Natriummolybdat wurde beschrieben. Eine bleiempfindliche Indikatorelektrode und eine Bezugselektrode mit KNO₃-Zwischenelektrolyt wurde in Kombination mit einem pH-Meter mit gedehnter Skala zur Erkennung des Titrationsendpunktes verwendet. Unter Standardisierung der Maßlösung gegen 1,5 mg Blei(II) werden genaue Resultate im Bereich von 1 bis 2 mg Blei erhalten: der absolute Fehler ist innerhalb ±3g Blei. Die relative Standardabweichung beträgt 0,05%. Das günstigste pH liegt im Bereich 5–5,5. In Wasser-Aceton oder -Methanol nimmt der Potentialsprung am Äquivalenzpunkt stark zu; gleichzeitig wird jedoch das Ansprechen der Elektroden sehr langsam. Die Titration von Blei ist möglich auch in Medien von Natriumnitrat oder/und Natriumperchlorat, was die Anwendung dieser Titration auf die Bleimikrobestimmung in bleihaltigen organischen Verbindungen nach Mineralisierung auf nassem Wege mit HNO₃ oder/und HClO₄ ermöglicht. Chlorid (bis zum 1000fachen Gehalt an Blei) und Bromid (100fache Menge) stören nicht. Iodid und Sulfat stören. Fünf bleihaltige organische Verbindungen wurden erfolgreich analysiert; die Standardabweichung des absoluten Fehlers betrug 0,1%.

     

In vivo tibial lead measurement to assess the health hazard of Venezuelan workers

Gamma-ray induced X-ray emission (GIXE) technique for elemental lead in-vivo tibial measurement using a large volume hyper pure germanium detector is presented with the most convenient source-sample-detector geometry. The system operates advantageously when several parameters are considered for a lower dead time operation. The detection limit (DL) is better that 3.5 μg/g of Ca. Results of in-vivo average tibial lead concentration for some of the monitored groups are: control 7 μg of lead per g of Ca; gasoline filling attendants 6 μg/g of Ca; custom office workers near the DL; industrial workers range from DL up to 84±3 μg/g of Ca, suggesting that lead accumulation in the bone does not represent a major health risk This revised version was published online in August 2006 with corrections to the Cover Date.     

In search of new lead compounds for trypanosomiasis drug design: A protein structure-based linked-fragment approach

Summary A modular method for pursuing structure-based inhibitor design in the framework of a design cycle is presented. The approach entails four stages: (1) a design pathway is defined in the three-dimensional structure of a target protein; (2) this pathway is divided into subregions; (3) complementary building blocks, also called fragments, are designed in each subregion; complementarity is defined in terms of shape, hydrophobicity, hydrogen bond properties and electrostatics; and (4) fragments from different subregions are linked into potential lead compounds. Stages (3) and (4) are qualitatively guided by force-field calculations. In addition, the designed fragments serve as entries for retrieving existing compounds from chemical databases. This linked-fragment approach has been applied in the design of potentially selective inhibitors of triosephosphate isomerase from Trypanosoma brucei, the causative agent of sleeping sickness.     

The composition of the plants’ extracts included in the herbal mixtures used to treat the parodontal disease

The methods of gas-liquid chromatography and mass-selective detection were used to determine the composition of the volatile fraction of plant extracts included in herb mixtures used for treating parodontal disease. The following marker substances were found: naphthalene derivatives for pot marigold, furan derivatives for trifid bur-marigold and burnet, and the specific terpenoids for licorice, milfoil, and wild marjoram. All the plants used in the herb mixture, except for pot marigold and cinquefoil, were analyzed (ion-exclusion chromatography) for the content of lower carboxylic acids. It was established that the specific acids for the wild marjoram and licorice are ascorbic and the succinic acids, respectively.     

Assessment uncertainty associated to the analysis of organic compounds linked to particulate matter of atmospheric aerosols

Current work has evaluated uncertainty associated to quantification of several organic compounds present in particulate matter of atmospheric aerosols, setting out the stages of analytical procedure that contribute most to the global uncertainty. Several sources of uncertainty have been identified, which were clustered into five main contributions: sampling, extraction, clean-up, derivatization and analysis. A discussion of the main contributions to the overall uncertainty is reported, allowing authors to locate the largest ones and plan future improvements. Combined uncertainties ranged between 10-18% (alkanes), 12-16% (PAHs), 10-18% (alcohols) and 9-21% (acids). The analytical procedure was validated by analysing a standard reference material (SRM1649a, urban dust). Also, the proposed method was applied to the analysis of four samples of particulate matter.     

A new method for the atomic-absorption determination of lead blended as lead alkyls in motor spirit

The total lead present as tetraethyl-lead (TEL) and tetramethyl-lead (TML) in motor spirit is determined by atomic-absorption spectrometry after decomposition by extraction with a mixture of mercaptoacetic acid and nitrous acid in the presence of hydrochloric acid.     

Replacement of chlorides with dicarboxylate ligands in anticancer active Ru(II)-DMSO compounds: a new strategy that might lead to improved activity

A series of new Ru(II)-DMSO complexes containing dicarboxylate ligands (dicarb), namely, oxalate (ox), malonate (mal), methylmalonate (mmal), dimethylmalonate (dmmal), and succinate (suc), have been synthesized and structurally characterized. These compounds were prepared from the known Ru(II)-Cl-DMSO anticancer complexes cis,fac-[RuCl2(DMSO-S)3(DMSO-O)] (1) and trans-[RuCl2(DMSO-S)4] (2) and from the chloride-free precursor fac-[Ru(DMSO-S)3(DMSO-O)3][CF3SO3]2 (3), with the aim of assessing how the nature of the anionic ligands influences the biological activity of these species. Basically, the investigated ligands can be divided into two groups. The reaction of either 1 or 2 with K2(dicarb) (dicarb = ox, mal, mmal) yielded preferentially the mononuclear species [K]fac-[RuCl(DMSO-S)3(eta2-dicarb)] (dicarb = mal, 6; mmal, 9; ox, 14) that contains a chelating dicarboxylate unit and a residual chloride. Likewise, when 3 was used as a precursor, the neutral mononuclear species fac-[Ru(DMSO-O)(DMSO-S)3(eta2-dicarb)] (dicarb = mal, 7; mmal, 10; ox, 16), which contains a DMSO-O ligand in the place of Cl-, was obtained. On the contrary, K2(suc) and K2(dmmal) yielded preferentially the dinuclear species [fac-Ru(DMSO-S)3(H2O)(mu-dicarb)]2 (dicarb = dmmal, 11; suc, 13), with two bridging dicarboxylate moieties. The two water molecules in anti geometry have strong intramolecular H-bonding with the non-coordinated oxygen atoms of the carboxylate groups. The solid-state X-ray structural data showed that the preferential binding mode of the investigated dicarboxylates, either bridging (mu) or chelating (eta2), is dictated mainly by steric reasons. Oxalate, unlike the other dicarboxylates, has also the bridging bis-chelate (eta4,mu) coordination mode available: this was found in the dinuclear species [{fac-RuCl(DMSO-S)3}2(eta4,mu-ox)] (15) and [{fac-Ru(DMSO-O)(DMSO-S)3}2(eta4,mu-ox)][CF3SO3]2 (17). We also isolated the unprecedented neutral metallacycle, [fac-Ru(DMSO-S)3(eta3,mu-ox)]4 (18), in which each oxalate unit has one unbound oxygen atom. The new complexes were thoroughly characterized by 1-D (1H and 13C) and 2-D (H-H- COSY and HMQC) NMR spectroscopy in solution and by IR spectroscopy in the solid state. The molecular structures of 10 compounds, 6-11, 13, 15, 17, and 18, were determined by X-ray crystallography. The behavior of selected complexes in aqueous solution was investigated by 1H NMR spectroscopy.     

A new approach to the formal classification of covalent compounds of the elements

A new approach to the classification of covalent compounds of the transition metals is presented. The classification introduces the concepts of valency number and ligand bond number as alternatives to the concepts of formal oxidation state and coordination number. Population density maps (MLX plots) which represent the covalent compounds of an element as a function of all the known ligands are exemplified for chromium; molybdenum and tungsten; and nickel, palladium and platinum. These plots are used to identify the major characteristic chemical properties of the elements including reactivity trends and reaction mechanisms.