Artificial blood: current status of hemoglobin solutions.

Attempts to develop a hemoglobin-based red cell substitute have spanned many decades, but no clinically useful product has been produced to date. The issues preventing clinical application primarily are ones of safety--not efficacy. Numerous animal studies have documented the efficacy of SFH. Although effective, the solution has limitations that have caused concern. Oncotic considerations limit the concentration of the infusate SFH to 6 to 8 g/dL, or half-normal. Owing to the loss of organic phosphate modulators of P50, such as 2,3-DPG, the P50 of SFH is typically between 12 and 14 mm Hg, which is also half the normal value. And finally, the intravascular half-life of SFH is too short, ranging only from 2 to 6 hr. Polymerization provides a means of correcting these limitations. The high oxygen affinity can be greatly diminished by covalent binding of pyridoxal-5'-phosphate to the N-terminal of the chains. Colloid osmotic pressure exerted by a protein solution is proportional to the number of discrete colloid particles. Through polymerization, the number of colloid particles is reduced, leading to a decrease in COP. Data show that this can be achieved in a reproducible fashion. The rate at which COP diminishes determines the yield of polymeric species, as well as their molecular weight distribution. Polymerization can be controlled to result in a yield of 75% to 85% polymers with a molecular weight distribution of 128 to 400 kd. The number average and the weight average molecular weights indicate that the large proportion of polymers represent the cross linking of two tetramers. The data that reflect the interaction of oxygen with poly-SFH-P indicate that the oxygen carrying function of hemoglobin has not been significantly altered by the chemical modifications. The binding coefficient of oxygen is unchanged. As anticipated, there is a loss of cooperativity (diminished Hill coefficient) between the hemoglobin chains, suggesting structural restrictions in the polymeric species because of cross linking. A reduced alkaline Bohr effect is the expected result, and data confirm this. Finally, some increase in oxygen affinity is to be expected with polymerization. This is indeed the case, although the P50 of poly-SFH-P is comparable to banked blood (18 to 22 mm Hg). To be clinically useful, a modified hemoglobin solution requires a reasonable shelf-life.(ABSTRACT TRUNCATED AT 400 WORDS)     

Effect of various stabilizing agents on Imperata cylindrica grass pollen allergen extract

BACKGROUND: Allergen extracts are unstable, heat labile or susceptible to proteases. Stability of allergen extracts is important for proper diagnosis and therapy of allergic disorders. OBJECTIVE: The present study was undertaken to determine the preservation and stabilization conditions of Imperata cylindrica (Ic) grass pollen extract. METHODS: The Ic extract was kept with 0.1 mepsilon-aminocaproic acid (EACA), 0.75 m sucrose, 5% glycerol, 0.03% human serum albumin (HSA) or 0.4% phenol for different time periods. The extracts were stored for 3, 6 and 12 months each at 4 degrees C, 4 degrees C with daily exposure to room temperature (RT) for 1 h, and RT. The quality of extracts was analysed by SDS-PAGE, Western blot, ELISA, ELISA inhibition and skin test. RESULTS: Extracts kept with EACA and sucrose retained most of the protein bands followed by glycerol as determined by SDS-PAGE and Western blot during all storage periods and conditions in comparison with standard extracts. The extracts kept with HSA, phenol and without preservative (WP) showed protein degradation below 33 kDa after 3 months storage at all conditions. However, a 67-kDa allergen was stable in these extracts. EACA extract required 75 to 120 ng of protein for 50% inhibition in IgE binding under different conditions, whereas standard extract required 70 ng for the same. ELISA also demonstrated high allergenic reactivity of EACA extract. ID test on allergy patients with EACA extract demonstrated same allergenic potency as that of standard extract. CONCLUSION: EACA is the best preservative/stabilizing agent of Ic pollen extract, followed by sucrose and glycerol. Ic extract kept with phenol, HSA and without preservative showed degradation within 3 months. EACA preserved extract is equally potent as that of standard extract up to 1 year's storage.     

Cadmium Mobilization by Monoaralkyl- and Monoalkyl Esters of meso-2,3-Dimercaptosuccinic Acid and by a Dithiocarbamate

Syntheses and relative cadmium mobilizing properties are described for three new monoaralkyl esters (HOOCCH(SH)CH(SH)COOR, where R = phenylethyl ((CH₂)₂C₆H₅), MPhEDMS; R = 3-phenylpropyl ((CH₂)₃C₆H₅), MPhPDMS; and R = 2-phenoxyethyl ((CH₂)₂OC₆H₅), MPhOEDMS) of meso-2,3-dimercaptusuccinic acid. These were prepared by the reaction of the corresponding alcohol with meso-2,3-dimercaptosuccinic acid (DMSA) in aqueous HCl. When administered intraperitoneally to cadmium-loaded mice at 0.50 mmol/kg/day for four consecutive days, all induced significant reductions in the whole body cadmium levels. MPhEDMS, 60%; MPhPDMS, 66%; and MPhOEDMS, 58% in comparison with control levels. At the same dosage monoisoamyl meso-2,3-dimercaptosuccinate (Mi-ADMS) and a dithiocarbamate, sodium N-benzyl-4-O-(β-D-galactopyranosyl)-d -glucamine-N-carbodithioate (BLDTC) induced reductions of 65% and 57%, respectively. Hepatic and renal cadmium were also depleted significantly, while brain cadmium levels were unchanged. These compounds induced a significant reduction in the cadmium levels of the spleen, and one, MPhOEDMS, produced a 10% decrease in pancreatic cadmium. The manner in which the later injections removed smaller fractions of the total body cadmium is consistent with a bodily distribution of these compounds by which they are concentrated primarily in the kidneys and the liver, with much smaller amounts reaching other organs. It is proposed that these compounds enter renal and hepatic cells through an anion transport system.     

Epidemiology of gastro-intestinal and respiratory tract diseases in rural areas of Varanasi (India)

The pattern of gastro-intestinal (GIT) and Respiratory tract (RTI) diseases in rural areas of the Varanasi district, U.P. (India) was studied from the outpatient (OPD) attendance between July 1985 to June 1986. Out of a total of new patients, 19.71% had respiratory diseases in contrast to 27.9% with GIT disease. A decreasing trend was observed in attendance rates as age advanced. The difference in age distribution of males and females was statistically significant. The attendance rates for GIT diseases were higher in female (29.16%) than in males (25.16%) whereas in RTI, the rates were higher in males (19.39%) than in females (18.77%). The attendance rate was found to be higher in rainy season (23.59%) followed by summer (14.50%). Diarrhoea (15.44%) and upper (URTI) respiratory tract infection (19.93%) occupied the major recorded episodes.     

Myopia and plasma cortisol

Estimation of plasma cortisol by fluorometric technique desorbed by Mallingly's was carried out in 56 cases, including 38 cases of myopia (19 cases of simple myopia and 19 cases of degenerative myopia) and 18 normal individuals. Urinary 17-keto steroids/24 hours was also estimated by Zimmermann technique in 12 out of the 56 cases, which include 8 cases of myopia (4 cases of simple myopia and 4 cases of degenerative myopia) and 4 normal individuals as control. Plasma cortisol level is lower in simple and degenerative myopia than in normal subjects, but on statistical analysis the difference was not of much significance. Patients with simple myopia with positive family history showed lower value of plasma cortisol than patients with simple myopia with negative family history, the difference was also statistically insignificant (P-0.1). In degenerative myopia patients, with and without family history, there was very little difference in plasma cortisol level and statistically highly insignificant (P-0.8). Urinary 17 keto steroid/24 hour values are lower in simple and degenerative myopia than in normal subjects.