Rapid cation-exchange high-performance liquid chromatographic procedure for the separation and quantitation of hemoglobins S, C, and O Arab in cord blood samples

A rapid method for separating and quantitating hemoglobin (Hb) variants in cord blood samples using cation high-performance liquid chromatography (HPLC) is described. The procedure is a modification of a previously published method, and uses a weak cation-exchange Brownlee-3CM column and Bis-Tris-KCN-Na acetate developers. A chromatogram can be completed in 10 minutes. The slow-moving variants, Hbs S, C, and O Arab, can be completely separated from each other and are identified by their elution times relative to Hb A. Hb E elutes as a shoulder on the descending side of Hb A, which is characteristic for this variant in this procedure. Differentiation between heterozygous, homozygous, and Hb X-beta+-thalassemia conditions is easily made.     

Characterization of normal and infarcted rat myocardium using a combination of small-animal PET and clinical MRI.

The combination of small-animal PET and MRI data provides quantitative in vivo insights into cardiac pathophysiology, integrating information on biology and morphology. We sought to determine the feasibility of PET and MRI for the quantification of ischemic injury in the rat model. METHODS: Fourteen healthy male Wistar rats were studied with 18F-FDG PET and cine MRI. Myocardial viability was determined in a transmural myocardial infarction model in 12 additional rats, using 18F-FDG PET and delayed-enhancement MRI with gadolinium-diethylenetriaminepentaacetic acid. All PET was acquired with a dedicated small-animal PET system. MRI was performed on a 1.5-T clinical tomograph with a dedicated small-animal electrocardiographic triggering device and a small surface coil. RESULTS: In normal rats, 18F-FDG uptake was homogeneous throughout the left ventricle. The lowest mean uptake of the 18F-FDG was found in the apical regions (79% +/- 6.0% of maximum) and the highest uptake was in the anterior wall (93% +/- 4.3 % of maximum). Myocardial infarct size as determined by histology correlated well with defects of glucose metabolism obtained with 18F-FDG PET (r = 0.89) and also with delayed-enhancement MRI (r = 0.91). Left ventricular ejection fraction in normal rats measured by cine MRI was 57% +/- 5.4% and decreased to 38% +/- 12.9% (P < 0.001) in the myocardial infarction model. CONCLUSION: Integrating information from small-animal PET and clinical MRI instrumentation allows for the quantitative assessment of cardiac function and infarct size in the rat model. The MRI measurements of scar can be complemented by metabolic imaging, addressing the extent and severity of ischemic injury and providing endpoints for therapeutic interventions.     

Effect of ileo-rectal anastomosis and post-valve T-caecum cannulation on growing pigs. 2. Blood variables and mineral balances.

In a long-term study nine ileo-rectally anastomosed (IRA) and seven post-valve T-caecum (PVTC)-cannulated pigs were compared with six intact pigs with regard to different blood variables, sodium and potassium retention and weights of selected organs. After surgery, apart from urea and K measured 13 weeks post-surgery, there were no differences in the blood variables between the PVTC-pigs and intact pigs. In IRA-pigs concentrations of creatinine (P < 0.01), Na (P < 0.001), base excess (P < 0.001), pH (P < 0.01) and bicarbonate (P < 0.001) in blood were lower than those in intact pigs. At 13 weeks after surgery the blood K concentration in IRA-pigs was higher (P < 0.001) than that in PVTC-pigs or intact pigs. At 6 weeks after surgery the blood urea concentration in IRA-pigs was higher (P < 0.001) than that in intact and PVTC-pigs. At 13 weeks after surgery the urea concentration in PVTC-pigs was higher (P < 0.001) than those in IRA-pigs or intact pigs. The Na (P < 0.01 11 weeks after surgery) and (P < 0.05 and P < 0.01 5 and 11 weeks after surgery respectively) balances in IRA-pigs were lower than those in intact animals. Na retention was negative for IRA-animals 11 weeks after surgery. Na and K retentions were similar in PVTC-pigs and in intact pigs. The urinary: faecal excretion of Na differed slightly between PVTC-animals and intact animals. At 13 weeks after surgery there were no differences in organ weights between the PVTC-pigs and intact animals. In the IRA-pigs the weights of the liver (P > 0.05), the kidneys (P > 0.05) and the adrenal glands were higher (P < 0.001) than those in the intact animals.     

Detection of circulating human platelet fragments by using monoclonal antibodies against platelet glycoprotein IIb-IIIa complex

A radioimmunoassay (RIA) for the detection of platelets and platelet fragments was developed. A sandwich of two monoclonal antibodies directed against the platelet-specific glycoprotein complex IIb-IIIa (GP IIb-IIIa) was used in this assay. A discontinuous 7.5-20% (v/v) albumin gradient was applied to separate platelets and their fragments of various sizes. In platelet suspensions fractionated in this way, we observed that particles smaller than normal platelets still carried the GP IIb-IIIa antigens. This procedure enabled us to detect platelet-derived particles in platelet-rich plasma from thrombocytopenic patients.     

Hb F-Xin-Su or A gamma I73(E17)Asp----His: a new slow-moving fetal hemoglobin variant

A slow-moving gamma chain variant was discovered in the cord blood of an infant born to parents of the Han nationality from Jiangsu. The variant, which migrated electrophoretically at alkaline pH between Hb A and Hb D, was characterized by an Asp----His substitution at gamma 73, while residues gamma 75 and gamma 136 were occupied by Ile and Ala, respectively. As the A gamma T-chain was also present, this baby has two types of abnormal gamma-chain. The variant was named Hb F-Xin-Su.     

Hb Las Palmas or alpha 2 beta 2(49)(CD8)Ser----Phe, a mildly unstable hemoglobin variant

A new hemoglobin variant with a Ser----Phe substitution at position beta 49(CD8) was discovered in two members of a family living in the Canary Islands, Spain. Detection was by polyacrylamide gel electrophoresis and by reversed phase high performance liquid chromatography. The variant, which constituted 43% and 45%, respectively, in the two heterozygotes, was slightly unstable. Its presence did not affect hematological values though there was a mild reticulocytosis.