Hereditary non-spherocytic haemolytic anaemia due to red blood cell glutathione synthetase deficiency in four unrelated patients from Spain: clinical and molecular studies.

In four unrelated patients with chronic haemolysis and markedly reduced red blood cell (RBC) glutathione (49.5%, 12.6%, 11.5% and 15% of the normal concentration respectively), a severe glutathione synthetase (GSH-S, EC 6.3.2.3) deficiency was found. One case exhibited a neonatal haemolytic anaemia associated with oxoprolinuria, but without neurological manifestations. The family study revealed GSH-S activity in both parents to be around half the normal level, a finding consistent with the presumed autosomal recessive mode of inheritance of this enzymopathy. Two cases exhibited a well-compensated haemolytic syndrome without anaemia or splenomegaly at steady state. One of these cases was diagnosed after an episode of acute haemolytic anaemia after fava bean ingestion. The remaining patient suffered from moderate to severe chronic non-spherocytic haemolytic anaemia and splenomegaly, and required occasional blood transfusion for a haemolytic crisis associated with drug ingestion. In this patient, the anaemia was corrected by splenectomy. In addition to GSH-S, a panel of 16 other RBC enzyme activities was also studied in all the patients. Hexokinase, aldolase, glucose-6-phosphate dehydrogenase and pyruvate kinase activities all increased; these increases were to be expected, given the rise in the number of circulating reticulocytes. In two patients, the incubation of RBCs with hydrogen peroxide revealed an enhanced production of malonyldialdehyde. DNA analysis showed a homozygous state for 656 A-->G mutation in patients 2 and 3. The GSH-S gene of patient 1, studied elsewhere, revealed an 808 T-->C. The GSH-S gene of patient 4 was not available for study. The present study demonstrates that GSH-S deficiency is also present in Spain and further supports the molecular and clinical heterogeneity of this enzymopathy

Mutation analysis in familial hypercholesterolemia patients of different ancestries: identification of three novel LDLR gene mutations.

Twelve familial hypercholesterolemia (FH) patients of different ancestries living in South Africa were subjected to mutation analysis of the low-density lipoprotein receptor (LDLR) gene. Nine different mutations were identified in 10 patients. Six of these, including the founder-related mutation C660X identified in two Lebanese patients, have previously been described in other FH patients with compatible genetic backgrounds, and/or in patients originating from countries where admixture is not uncommon. Characterization of an abnormal electrophoresis pattern detected in exon 4 of the LDLR gene by heteroduplex single-strand conformation polymorphism (HEX-SSCP) analysis, revealed a novel G deletion at codon 185 (617delG) which resulted in a downstream stop codon. Two of the new mutations identified resulted in amino acid substitutions and were designated R57C and Q357P.

Autologous stem cell transplantation for poor prognosis Hodgkin's disease in first complete remission: a retrospective study from the Spanish GEL-TAMO cooperative group.

Although more than 50% of Hodgkin's disease patients are cured with conventional chemotherapy, many will relapse and eventually die from their disease. Many efforts have been made to identify poor prognostic factors that could be useful in selecting high-risk patients in 1st CR who may benefit from high-dose chemo/radiotherapy. However, the role of early transplantation in 1st CR remains unclear. We have retrospectively analyzed the results obtained with this procedure in 22 hospitals belonging to the Spanish GEL/TAMO cooperative group. Twenty-seven patients, of whom 19 were males, underwent autologous transplantation for Hodgkin's disease in 1st CR between January 1987 and January 1996. Remission had been achieved after one (n = 22) or two (n = 5) lines of treatment. Twenty-four patients had advanced stage disease, 12 patients bulky mediastinal disease, nine bone marrow involvement and 18 had extranodal disease. Peripheral blood was used as the source of hematopoietic stem cells in 15 patients, BM in nine, and both in three. All but three patients received chemotherapy-based conditioning regimens (16 CBV, four BEAM and four BEAC), while three were conditioned with CY and TBI. There were no transplant-related deaths. Median (range) times to recover >0.5 x 10(9)/l neutrophils and >50 x 10(9)/l platelets were 14 (8-56) days and 16 (8-240) days, respectively. With a median follow-up of 30 (8-66) months, 21 patients are alive and in continuous CR. Four patients who relapsed after transplant at 8, 17.5, 22 and 26 months achieved a second CR with conventional chemotherapy; one patient relapsed 92 months post-transplant and died 5 months afterwards. Another patient died 30.5 months post-transplant from a secondary malignancy. In conclusion, high-dose therapy in poor prognosis Hodgkin's disease in 1st CR was well tolerated with no transplant-related mortalities. Although the follow-up of this series is relatively short, our results seem promising. Nevertheless, late relapses can occur, and the role of this procedure vs conventional treatment in very high-risk patients should be assessed in prospective randomized studies.

Nonradioactive multiplex PCR screening strategy for the simultaneous detection of multiple low-density lipoprotein receptor gene mutations.

We have developed a rapid, nonradioactive screening test enabling the simultaneous analysis of three low-density lipoprotein receptor (LDLR) gene mutations (D154N, D206E, and V408M), which together account for familial hypercholesterolemia (FH) in approximately 90% of the South African Afrikaner population. The assay is designed so that FH patients, negative for these founder-related mutations (found in descendants of European settlers), subsequently can be screened for unknown mutations in the mutation-rich exon 4 of the LDLR gene. Our screening assay consists of two steps: (1) multiplex allele-specific PCR amplification of exons 4 and 9, and (2) simultaneous analysis of single- and double-strand conformational polymorphisms in exon 4 by vertical electrophoresis on low cross-linked polyacrylamide gels. The simplicity, specificity, and versatility of the multiplex assay makes it an ideal system for routine screening of FH mutations in large population samples.

A modified internal rotation stretching technique for overhand and throwing athletes.

Strength, stretching, and rehabilitation methods for the shoulder have been previously described and have been universally applied; nevertheless, many throwing athletes continue to develop overuse injuries. The pitching process tends to increase external rotation and limit internal rotation of the shoulder joint. Our technique involves a modified stretching method of the posterior shoulder musculature. The athlete lies prone with the elbow flexed 90 degrees. With the shoulder abducted 90 degrees, in neutral flexion/extension, and 90 degrees or more of internal rotation, the scapula protrudes posteriorly. By depressing the inferior angle of the scapula toward the thoracic wall, the infraspinatus muscle and posterior joint capsule are effectively isolated and stretched. Manual stabilization of the scapula onto the chest wall transfers the internal rotation movements to the glenohumeral joint, as opposed to sharing the movement with the scapulothoracic articulation. This method improves the efficacy of the internal rotation stretching exercise for the glenohumeral joint. Such an addition to traditional stretching methods may increase the efficiency of the least effective component. We conclude that this modification to traditional programs should be an integral part of the training and rehabilitation program of any athlete requiring near maximal performance of the shoulder.

Emergence of quinolone-resistant Escherichia coli bacteremia in neutropenic patients with cancer who have received prophylactic norfloxacin.

Between January 1988 and December 1992, 35 episodes of Escherichia coli bacteremia were identified in a series of 230 cases of bacteremia in neutropenic patients with cancer. Thirteen episodes (37%) were due to quinolone-resistant strains. Minimal inhibitory concentrations of norfloxacin ranged from 16 micrograms/mL to 128 micrograms/mL, and those of ciprofloxacin from 8 micrograms/mL to 64 micrograms/mL. The incidence of bacteremia due to quinolone-resistant E. coli increased from zero episodes per 1,000 hospital admissions in 1988 to four episodes per 1,000 admissions in 1992 (P = .018). To identify risk factors for quinolone-resistant E. coli bacteremia, we compared episodes of quinolone-resistant and quinolone-susceptible E. coli bacteremia. Among the variables analyzed, prophylaxis with norfloxacin was the only factor significantly associated with the development of quinolone-resistant E. coli bacteremia; 13 of 13 patients with bacteremia due to resistant strains received norfloxacin, whereas only one (5%) of 22 patients with bacteremia due to susceptible strains did (P < .001). According to our data, neutropenic patients with cancer who receive fluoroquinolone prophylaxis may be at risk of developing E. coli bacteremia due to quinolone-resistant strains.

[Alfa-2b interferon in the treatment of thrombocytosis associated to chronic non leukemic myeloproliferative syndromes]. / Interferón alfa-2b en el tratamiento de la trombocitosis asociada a síndromes mieloproliferativos crónicos no leucémicos.

BACKGROUND: The effect of interferons in the correction of thrombocytosis in chronic myeloproliferative syndromes is well known. In this study the efficacy of alpha-2b interferon in a regimen of induction followed by a phase of sequential maintenance to progressively decreasing doses was evaluated with the aim of knowing the minimum doses necessary to maintain response. METHODS: The response to treatment with alpha-2b interferon was prospectively studied in a group of 37 patients with chronic myeloproliferative syndromes with associated thrombocytosis (excluding chronic myeloid leukemia). Likewise, the toxicity of the treatment was analyzed. RESULTS: Sixty-seven percent of the patients responded (platelets lower than 600 x 10(9)/1) to the daily administration of 3 or 5 MU of interferon. Forty percent of the patients who responded to the daily schedule of administration maintained the response upon receiving 3 doses weekly for 4 months. Half of the 8 patients who received 2 weekly doses of interferon for 4 months continued maintaining the responses. Only two of the 4 patients who received one sole weekly dose during the following 4 months maintained the response. Only one of the 37 patients who initiated treatment underwent progression of the symptoms present at the beginning of the study. Toxicity was high and was the cause of 12 discontinuations of treatment (32% of the patients) during the daily treatment phase (9 patients) or during maintenance of 3 weekly doses (3 patients). No toxicity was observed in the schedule of one or two weekly doses. CONCLUSIONS: Alpha-2b interferon is effective in the treatment of thrombocytosis of the chronic myeloproliferative syndromes (excluding chronic myeloid leukemia) when administered daily and is ever less so when the doses are spaced at 3, 2 or 1 week. The toxicity of interferon treatment is high when administered at affective doses.

Yeasts associated with Vienna sausage packaging.

A total of 123 representative yeast isolates from a previous study of a Vienna sausage processing plant were identified according to conventional methods and long-chain fatty acid analyses. The most prevalent isolates belonged to the genera Candida and Debaryomyces. Other genera encountered were Rhodotorula, Yarrowia, Pichia, Galactomyces, Cryptococcus, Trichosporon and Torulaspora.

Hairy cell leukemia and autoimmune hemolytic anemia.

A case of hairy cell leukemia (HCL) presenting as autoimmune hemolytic anemia (AHA) is described. A 40-year-old woman presented with severe hemolytic anemia. The morphological and immunological studies of bone marrow and spleen revealed a hairy cell leukemia. Although autoimmune diseases are a well known complication of HCL this is the first description of AHA as a complication of HCL.

[Vindesine, CCNU, high-dosage ara-C, and prednisolone (VINAP regimen) in the treatment of relapsing or refractory non-Hodgkin's lymphomas]. / Vindesina, CCNU, dosis altas de Ara-C y prednisolona (régimen VINAP) en el tratamiento de los linfomas no hodgkinianos en fase de recaída o refractarios.

Results of second line chemotherapy schedules to treat refractory lymphoma have usually been poor. In this study we have treated 21 patients with advanced non-Hodgkin's lymphoma, usually heavily pretreated, with VINAP regimen. This original four drug chemotherapy combination included: Vindesine, 2 mg/m2 iv on days 1 and 2; CCNU, 40 mg/m2 oral on days 3 and 4; Cytosine arabinoside, 2.4 g/m2 iv on day 3 to 6 and methyl-Prednisolone, 80 mg/m2 on days 1 to 6. Sixteen patients (76%) showed response, including 5 (23%) who achieved a complete remission (CR). Eight patients achieved a partial remission (PR), and two patients obtained an objective response. Although the responses to VINAP regimen were dramatic and rapid in onset, usually they were of short duration except in cases of lymphosarcoma cell leukaemia. The median duration of response for patients with CR was 42 weeks and for PR 11 weeks. Toxicity was acceptable, including predictable myelosuppression, frequent mucositis and occasional polyneuritis. Neither central nervous problems nor conjunctivitis or dermatitis had been seen.