ABSTRACT
BACKGROUND: Identification of RBC pyruvate-kinase (PK) gene mutations by polymerase chain reaction (PCR) and single strand conformation polymorphism (SSCP) followed by PK gene sequencing in positive cases has been assessed and the results obtained with a preliminary study of 15 unrelated patients of Spanish origin are presented. PATIENTS AND METHODS: Patients have been classified into two different groups: group 1, propositus (15 cases), and group 2, relatives of the patients included in group 1 (10 males and 5 females). In group 1, a PCR was followed by SSCP and sequencing, and in group 2, the PCR was followed by digestion with specific restriction endonucleases (PCR-ER). RESULTS: Group 1: from 15 patients included in the study 2 were identified as homozygous, 4 as heterozygous and 9 as compound heterozygous. In this group, were identify 26 affected alleles with 11 different mutations: T1456 10 alleles (38.6%), T721 3 alleles (11.6%), A1010, C514, C1015 and T1223 2 alleles (7.7%), and C1070, A1291, T1508, A1595 y T1675 one allele. Relatives from 8 out of 15 patients from group 1 showed the following pattern: homozygous (one case), heterozygous (10 cases), compound heterozygous (2 cases) and normal (2 cases). CONCLUSIONS: SSCP procedure followed by direct gene sequencing in positive cases is fast and simple enough to allow the identification of PK deficient variants, avoiding the need of biochemical characterisation of semipurified deficient enzyme, which is more cumbersome and time consuming. In addition, the PCR-ER method is a very useful tool for screening of the most frequent molecular variants, as well as, for the detection of the carrier condition of this enzymopathy (family studies).
Subject(s)
Anemia, Hemolytic/genetics , Erythropoiesis/genetics , Pyruvate Kinase/deficiency , Pyruvate Kinase/metabolism , Amino Acids/genetics , Anemia, Hemolytic/enzymology , Chronic Disease , Female , Genotype , Hematology , Humans , Male , Nucleotides/genetics , Point Mutation/genetics , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , Sequence Analysis, DNA , Societies, Medical , SpainSubject(s)
Amyloidosis/drug therapy , Doxorubicin/analogs & derivatives , Hodgkin Disease/complications , Adult , Amyloidosis/etiology , Doxorubicin/metabolism , Doxorubicin/therapeutic use , Edema/drug therapy , Edema/etiology , Fatigue/etiology , Humans , Male , Nephrotic Syndrome/drug therapy , Nephrotic Syndrome/etiology , Proteinuria/drug therapy , Proteinuria/etiology , Serum Amyloid A Protein/analysis , Serum Amyloid A Protein/metabolismSubject(s)
Antineoplastic Agents/therapeutic use , Hematopoietic Stem Cell Transplantation/methods , Interferon-alpha/therapeutic use , Multiple Myeloma/therapy , Registries/statistics & numerical data , Antineoplastic Agents/adverse effects , Combined Modality Therapy , Humans , Interferon-alpha/adverse effects , Multiple Myeloma/mortality , Recurrence , Spain/epidemiologySubject(s)
Leukemia, Neutrophilic, Chronic , Chromosome Aberrations , Clone Cells/pathology , Disease Progression , Humans , Leukemia, Neutrophilic, Chronic/genetics , Leukemia, Neutrophilic, Chronic/pathology , Multiple Myeloma/pathology , Myelodysplastic Syndromes/pathology , Neoplasms, Multiple Primary/pathology , Neoplastic Stem Cells/pathology , Neutrophils/physiology , Paraproteinemias/pathologyABSTRACT
A 70-yr-old man with multiple myeloma IgD developed a plasma cell leukemia producing a serum IgD monoclonal peak and lambda light chains in the urine. When the serum and the urine were heated at 56 degrees C for 30 min both monoclonal bands disappeared. The precipitate failed to redissolve on heating to 100 degrees C. Ion exchange chromatography with a linear gradient of phosphate buffer, pH 8, 0.020-0.300 mol/l and column electrofocusing showed that the serum pyroglobulin was eluted with buffer concentration between 0.040-0.125 mol/l and had an isoelectric point of 5.02, while the pyroglobulin of the urine was eluted with 0.020-0.033 mol/l and had a pI = 7.16. The serum and urine pyroglobulins had a total antigenic community with their correspondent purified proteins. The isolated lambda chains did not change when they were heated at 56 degrees C for 30 min, nevertheless, the heated purified IgD shows some changes in its isoelectric point, molecular mass and antigenicity. These changes in the purified IgD suggest that the pyroprecipitability could be due to conformational features.
Subject(s)
Immunoglobulin D , Leukemia, Plasma Cell/metabolism , Paraproteins/metabolism , Pyroglobulins/metabolism , Aged , Chemical Phenomena , Chemistry, Physical , Electrophoresis , Hot Temperature , Humans , Immunoassay , Immunoglobulin lambda-Chains , Isoelectric Focusing , Leukemia, Plasma Cell/etiology , Leukemia, Plasma Cell/immunology , Male , Molecular Weight , Multiple Myeloma/complications , Pyroglobulins/urineABSTRACT
Six cases of constitutional aplastic anemia (Fanconi's anemia), including four boys and two girls who were between 6 and 10 years of age, are presented. Congenital abnormalities were observed in all the patients, especially skin pigmentations, renal anomalies, and growth retardation. The initial symptoms were insidious, mainly related to the anemia. All patients were anemic and thrombocytopenic, but only three were leukopenic. Five cases showed increased levels of fetal hemoglobin, which was marked in two. Bone marrow obtained by aspiration and/or biopsy was hypocellular in all cases. Chromosomal studies showed breaks, ring- or cross-chromosomes, chromosome exchanges, and endoreduplications. Five cases were treated with androgens and a good response was obtained in all of them. Four patients died, and the remaining two are alive and well. The clinical and biological features of these patients are in accordance with those of previous reports. The importance of the androgen therapy is stressed; androgen dependence was observed in all cases.
