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The usefulness of the Coloscreen VPI test kit in occult blood detection has been examined and compared with our current method. The results show the Coloscreen procedure to be a simple, sensitive method that is much less subject to vegetable peroxidase interference than are other available screening procedures.
We describe an improved colorimetric method for assays of total and direct bilirubin in serum. Bilirubin reacts with diazotized sulfanilic acid in an acidic medium to form a blue azopigment. Total bilirubin is assayed in the presence of reaction accelerators (caffeine, urea, and citric acid), direct bilirubin in their absence. The azo compound so formed is read at the same wavelength (570 nm) in both assays. A sample blank is run in parallel. Standard curves are linear for total and direct bilirubin concentrations up to 513.0 and 256.5 mumol/L, respectively. The method is characterized by (a) use of the same protocol for both assays, i.e., a one-step procedure with short reaction time (5 min at room temperature), and (b) use of a single working solution, which, refrigerated, is stable for one month. The method is reliable, yields results that compare closely with those of the classical Jendrassik--Gróf method, is suitable for routine use, and lends itself to automation.
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The calculations of the electronic structure of fifth-coordinated ferroporphyrin-imidazole complexes modeling alpha- and beta-subunits in desoxyhemoglobin and desoxymyoglobin are made using the interative extended Hückel method. The features of the electronic structure of the model complexes resulting from the stereochemical differences of the active site are studied and compared. Theoretical calculations of the Mössbauer parameters of model complexes are made and compared with experimental data. The results show that basic features of the Mössbauer parameters are qualitatively confirmed by the theoretical analysis. The necessity of the accounting of the stereochemical and electronic structure nonequivalence of nonidentical subunits in tetrameric desoxyhemoglobins during the Mössbauer spectra approximation is also confirmed.
In a coastal population in whom anaemia was common, two randomised controlled trials were undertaken to investigate the effectiveness of treating iron deficiency anaemia at a dispensary and at primary schools. For anaemic adult villagers treated at a dispensary, one dose of tetrachlorethylene for hookworm infection and a once per week visit to collect medicines were found to be satisfactory. A small but significant increase in haemoglobin level was produced by four weeks oral iron therapy, but this was only maintained after seven months by the group that had initially also received tetrachlorethylene. In children (5-14 years) no significant rise in haemoglobin level was obtained by using oral iron and/or TCE, either at the dispensary or at the primary schools. This suggests that malaria was a more potent cause of anaemia in these children.
It has been established that one of the mechanisms of lipid peroxidation intensification is mediated by metals of variable valency, hemin compounds and phospholipids carried into the vitreous body by blood.
The barrier properties of erythrocyte membranes were studied under freezing-thawing in cryopreservatives based on glycerol or polyethylene glycol Mm 1500 Da (PEG-1500) (at cryoprotectant concentrations from 0.5 to 30%). The state of the membrane barrier function in reconstituted and native erythrocytes subjected to rapid (200-400 degrees C per min) freezing has been registered by spectrophotometry, flame photometry, and radiolabel analysis as judged by labeled compound release (hemoglobin, K+, [14C]sucrose, 86Rb+). It is demonstrated that the effective cryoprotection of erythrocyte membranes in reconstituted erythrocytes is attained at cryoprotectant concentrations in the medium of 0.5-1% or in erythrocytes of 8-30%.
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Resonance Raman spectra of the ferrous CO complex of cytochrome P-450cam have been observed both in its camphor-bound and free states. Upon excitation at 457.9 nm, near the absorption maximum of the Soret band, the ferrous CO complex of the camphor-bound enzyme showed an anomalously intense Raman line at 481 cm-1 besides the strong Raman lines at 1366 and 674 cm-1 for the porphyrin vibrations. The Raman line at 481 cm-1 (of the 12C16O complex) shifted to 478 cm-1 upon the substitution by 13C16O and to 473 cm-1 by 12C18O without any detectable shift in porphyrin Raman lines. This shows that the line at 481 cm-1 is assignable to Fe-CO stretching vibration. By the excitation at 457.9 nm, a weak Raman line was also observed at 558 cm-1, which was assigned to the Fe-C-O bending vibration, because it was found to shift by -14 cm-1 on 13C16O substitution while only -3 cm-1 on 12C18O substitution. These stretching and bending vibrations of the Fe-CO bond were not detected with the excitation at 413.1 nm, though the porphyrin Raman lines at 1366 and 674 cm-1 were clearly observed. When the substrate, camphor, was removed from the enzyme, the Fe-CO stretching vibration was found to shift to 464 cm-1 from 481 cm-1, while no detectable changes were found in porphyrin Raman lines. This means that the bound substrate interacts predominantly with the Fe-CO portion of the enzyme molecule.
The objective of the present study was to obtain information about the duration of tissue hypoxia in patients with iron deficiency anemia. That fact is of importance for the determination of the duration of maintenance iron therapy. The level of 2,3-diphosphoglycerate was studied during the treatment, after the correction of anemic syndrome and after 60-day out-patient department treatment. The data obtained revealed that the level of 2,3-diphosphoglycerate was considerably elevated, as compared with the norm, before the treatment. After the active treatment and correction of anemic syndrome it was decreased, but remaining above the norm. By the 60th day of the out-patient department treatment the decrease continued and the level of 2,3-diphosphoglycerate approached the norm.
Serum ferritin estimation was used to determine the iron stores in 55 patients with severe iron deficiency anaemia in pregnancy. The response to oral or intravenous iron therapy was monitored in 18 of these patients. The results in all cases indicated deficient iron stores. Replenishment of iron stores was significantly greater in those patients treated with parenteral iron. This may be the treatment of choice for severe iron deficiency anaemia during pregnancy.
Free iron in the circulation in contrast to iron complexed with protein or carbohyrdate, is intensely toxic. The annual morbidity and mortality from accidental overdose with iron salts is significant and has directed attention to the evaluation of safer forms of therapy. Comparative studies between oral ferrous sulphate and iron polymaltose in rats established an LD100 of 350 mg/kg for the salt, whereas neither morbidity nor mortality could be produced in doses exceeding 1 000 mg/kg for the complex. Bio-availability of iron from salt and complex for haemoglobin synthesis was then compared both in rats and in man with a twin-isotope technique. In experimental animals and in the human studies, venesected individuals were used to reproduce the iron deficiency state where treatment would be indicated. It was concluded that bioavailability is comparable for therapeutic doses of ferrous sulphate and iron polymaltose in iron-deficient subjects. The marginally lower absorption of complex was offset by a greater degree of gastrointestinal tract tolerance; the wide margin of safety may be of importance in reducing the danger of accidental iron overdose.
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