Safety and efficacy of therapeutic angiogenesis as a novel treatment in patients with critical limb ischemia.

BACKGROUND: In some patients with critical limb ischemia (CLI) the possibility of revascularizing treatment does not exist. In this case therapeutic angiogenesis (TA) using autologous endothelial progenitor cell (EPC) transplantation could be an alternative. The objective of our study was to evaluate the safety and efficacy of TA using EPC. METHODS: Twenty-eight patients with CLI who were not candidates for surgical or endovascular revascularization were included in a prospective study. To mobilize EPCs from the bone marrow, granulocyte colony-stimulating growth factor was injected subcutaneously at doses of 5 microg/kg/day for 5 days. Apheresis was performed, obtaining 50 mL of blood with a high rate of EPCs (CD34(+) and CD133(+) cells were counted). EPCs were implanted in the ischemic limb by intramuscular injections. Primary end points were the safety and feasibility of the procedure and limb salvage rate for amputation at 12 months. Other variables studied were improvement in rest pain, healing of ulcers, ankle-brachial pressure index (ABI), and digital plethysmography. All procedures were done pretreatment and every 3 months for a year on average. Postransplantation arteriography was done in selected cases. RESULTS: No adverse effects were observed. Mean follow-up was 14 months. Before treatment, mean basal ABI was 0.35+/-0.2 and at 18 months postimplantation, 0.72+/-0.51 (p=0.009). There was a mean decrease of five points in pain scale: basal 8.7+/-1, after TA 3.8+/-2.9 (p=0.01). Seven patients required major amputation. Kaplan-Meier analysis revealed a limb salvage rate of 74.4% after 1 year. CONCLUSION: Implantation of EPCs in CLI is a safe alternative, improves tissue perfusion, and obtains high amputation-free rates. Nevertheless, this is a small cohort and results should be tested with long randomized trials.

Rentabilidad de las muestras respiratorias pediátricas en la red centinela de vigilancia de la gripe en las Islas Baleares / Diagnostic yield of paediatric respiratory samples in the Balearic Islands Sentinel Influenza Surveillance Network

Introducción: La gripe es una enfermedad mundialmente vigilada a través de las redes centinela (RC) que predicen el comportamiento epidemiológico y comunican los datos clínicos y virológicos. Objetivos: Evaluar la rentabilidad de las muestras respiratorias pediátricas en la RC de las Islas Baleares en las últimas 5 temporadas gripales. Material y pacientes: Se tomó un frotis faríngeo para cultivo vírico de los pacientes que cumplían los criterios de gripe. Se inocularon las muestras en la línea celular MDCK. Se revisaron los datos epidemiológicos de cada paciente con cultivo positivo para los virus gripales A y B. Resultados: Durante el período de estudio se analizaron 338 frotis faríngeos de la RC. De éstos, 65 (19,3%) pertenecían a menores de 14 años. El 44,6% de estas muestras fueron positivas frente al 39,1% de las muestras de adultos. El virus gripal A se aisló en 24 muestras pediátricas (82,7%) y el virus gripal B se aisló en 5 muestras (17,3%). La edad media de los pacientes positivos de la RC fue de 8,5 años. En el grupo de 0 a 4 años sólo fueron positivos 3 pacientes (10,3%) y en el grupo de 5 a 14 años fueron positivos 26 pacientes (89,7%). Conclusiones: A pesar de que los pediatras sólo representan el 22% de los médicos de la RC y aportan el 19,3% de las muestras, el porcentaje de positividad y rentabilidad de éstas es superior al de la población adulta (AU)

Introduction: Influenza disease is subjected to surveillance by national networks (RC) that predict the epidemic behaviour by reporting clinical and virological data. Objectives: To evaluate the effectiveness of the paediatric respiratory samples in the Balearic Islands RC in the last five epidemic seasons. Material and patients: A breath sample was taken from paediatric patients in the RC who had flu symptoms. The samples were inoculated in the MDCK cell line. We reviewed the epidemiological data of patients with a culture positive to influenza A and B. Results: A total of 338 pharyngeal swabs from the RC were analysed during the study period. Of these, 65 (19.3%) belonged to <14 years old patients, and 44.6% of the samples were positive as opposed to 39.1% of adult respiratory samples. The influenza A virus was isolated in 24 paediatric samples (82.7%) and the influenza B virus in 5 (17.3%). The mean age of the paediatric patients of the RC who were positive was 8.5 years. Only 3 patients in the 0–4 year old group were positive (10.3%) and 26 patients (89.7%) in the 5–14 years old group. Conclusions: In spite that paediatricians represented only 22% of the RC doctors and obtain the 19.3% of all respiratory samples, the percentage and effectiveness of these is higher that obtained in the adult population (AU)

[Diagnostic yield of paediatric respiratory samples in the Balearic Islands Sentinel Influenza Surveillance Network]. / Rentabilidad de las muestras respiratorias pediátricas en la red centinela de vigilancia de la gripe en las Islas Baleares.

INTRODUCTION: Influenza disease is subjected to surveillance by national networks (RC) that predict the epidemic behaviour by reporting clinical and virological data. OBJECTIVES: To evaluate the effectiveness of the paediatric respiratory samples in the Balearic Islands RC in the last five epidemic seasons. MATERIAL AND PATIENTS: A breath sample was taken from paediatric patients in the RC who had flu symptoms. The samples were inoculated in the MDCK cell line. We reviewed the epidemiological data of patients with a culture positive to influenza A and B. RESULTS: A total of 338 pharyngeal swabs from the RC were analysed during the study period. Of these, 65 (19.3%) belonged to <14 years old patients, and 44.6% of the samples were positive as opposed to 39.1% of adult respiratory samples. The influenza A virus was isolated in 24 paediatric samples (82.7%) and the influenza B virus in 5 (17.3%). The mean age of the paediatric patients of the RC who were positive was 8.5 years. Only 3 patients in the 0-4 year old group were positive (10.3%) and 26 patients (89.7%) in the 5-14 years old group. CONCLUSIONS: In spite that paediatricians represented only 22% of the RC doctors and obtain the 19.3% of all respiratory samples, the percentage and effectiveness of these is higher that that obtained in the adult population.

In hereditary hemochromatosis, red cell apheresis removes excess iron twice as fast as manual whole blood phlebotomy.

The current treatment of hereditary hemochromatosis (HH) consists of performing periodic manual whole blood phlebotomies. Erythroapheresis (EPH) is considered to be an alternative procedure if the classic treatment is contra-indicated. A prospective study of 13 consecutive cases of HH were included in a periodic EPH program with the aim of assessing the efficacy, feasibility, and tolerability of EPH in the treatment of HH by induction and maintenance. Iron depletion (ferritin <20 microg/l) was achieved in all patients after a mean of 6.7 +/- 2.9 months of treatment and a mean of 13.5 +/- 7.2 EPH sessions. The procedure was well tolerated and there were no complications. After a follow-up period of 10.5 +/- 6.6 months, only four patients have required further maintenance sessions beyond 6 months after completing the induction therapy. The efficacy, speed, tolerability, and more favorable schedule of an EPH program facilitate treatment of HH.

Distribution of HFE C282Y and H63D mutations in the Balearic Islands (NE Spain).

The HFE gene contains two main missense mutations: C282Y and H63D. Individuals with these mutations carry a risk of developing hereditary haemochromatosis (HH). The common form of this disease is due to homozygosity for the C282Y mutation. Population studies have shown the variation of the prevalence of these mutations in different countries and ethnic groups. The purposes of this current study were to determine the prevalence of the C282Y and H63D mutations in the Balearic Islands and the genotypic characterization of patients diagnosed with HH, as well as those with iron overload and liver diseases. A total of 1330 Balearic chromosomes were analyzed. The results showed that the populations of the Balearic Islands were not homogeneous. No C282Y carriers were observed in a group of descendants of Majorcan Jews (Chuetas) and the frequency was very low in Minorca (1.2%) in comparison with the other islands of Majorca (4.7%) and Ibiza (6.5%). The carrier frequency of the H63D mutation was similar in the three islands and very high (43.1%) in the descendants of Majorcan Jews. The study of patients was carried out in 129 individuals. The homozygous C282Y genotype was the principal one involved in hereditary haemochromatosis (90%), whereas the other HH patients were C282Y/H63D compound heterozygous and H63D homozygous.

Two outbreaks of mumps in children vaccinated with the Rubini strain in Spain indicate low vaccine efficacy.

This paper reports a descriptive study of two outbreaks of mumps in Spanish towns in 1998 and an assessment of the efficacy of the vaccine used. It provides evidence of the low level of protection against the mumps virus that has been observed previously

Long-term storage at -80 degrees C of hematopoietic progenitor cells with 5-percent dimethyl sulfoxide as the sole cryoprotectant.

BACKGROUND: Hematopoietic progenitor cells (HPCs) can be cryopreserved and stored below -120 degrees C in liquid nitrogen or at -80 degrees C in mechanical freezers. STUDY DESIGN AND METHODS: The feasibility of long-term storage of HPCs at -80 degrees C was investigated. The studies included a comparison of 5- and 10-percent dimethyl sulfoxide (DMSO) as cryoprotectant at various lengths of storage time. Mononuclear cell (MNC) recovery and viability and colony-forming unit-granulocyte-macrophage (CFU-GM) and burst-forming unit-erythroid (BFU-E) recovery assays were performed. The peripheral blood HPCs of 24 consecutive patients included in the program of autologous transplantation were studied. RESULTS: The MNC viability decreased progressively with the length of time from cryopreservation, reaching 32 percent after 31 months of storage. The recovery rates of CFU-GM and BFU-E also decreased progressively with the duration of frozen storage, to 50 and 43.5 percent, respectively, after 12 months and to 0 percent (both) after 24 months. At 6 months of storage, MNC viability was 80 percent, and CFU-GM and BFU-E recovery was 63.5 and 80.5 percent, respectively. There were no differences between MNCs cryopreserved with 5- or 10-percent DMSO in terms of cell viability. There were no differences between CFU-GM recovery or BFU-E recovery from HPCs cryopreserved in 5- or 10-percent DMSO. Patients given HPCs stored in these conditions for periods ranging between 123 and 202 days showed a complete and rapid hematologic recovery. CONCLUSION: HPCs can be cryopreserved at -80 degrees C with 5-percent DMSO and stored at -80 degrees C no longer than 6 months. A 5-percent DMSO concentration is comparable to a with 10-percent concentration in terms of recovery and MNC viability.

Imaging features of type-B Niemann-Pick disease.

We report two cases of a mild form of type-B Niemann-Pick disease manifesting as an adult-onset chronic non-neuropathic clinical picture. Femoral T1- and T2-weighted low-intensity non-enhancing coarse bone marrow pattern was evident on femoral MR associated with splenic hypodense nodule(s) on abdominal CT. The role of imaging is discussed in relation to current techniques of confirmation of this entity based on demonstration of lipid-laden cells within marrow aspirates (which are often sea-blue histiocytes) and sphingomyelinase assay of cultured skin fibroblasts.

Cryopreservation of hematopoietic progenitor cells with 5-percent dimethyl sulfoxide at -80 degrees C without rate-controlled freezing.

BACKGROUND: Cryopreservation of hematopoietic cells with the rate-controlled method is used in the majority of centers. In recent years, there has been a trend toward the simplification of the process. STUDY DESIGN AND METHODS: A simplified method for cryopreservation was developed with 5-percent dimethyl sulfoxide (DMSO) as the sole cryoprotectant without rate-controlled freezing. Experiments were done with progressive concentrations of DMSO, ranging from 0 to 10 percent. With DMSO concentrations from 5- to 10-percent, the best recovery and viability for hematopoietic progenitor cells were observed. Hematopoietic progenitor cells with plasma and 5-percent DMSO were frozen and stored in a -80 degrees C mechanical freezer. Ten patients with solid and hematologic malignancies underwent transplantation with autologous hematopoietic progenitor cells. RESULTS: The median number of transfused mononuclear cells and CD34+ cells was 3.70 (3.1-8.2) x 10(8) per kg and 1.70 (0.8-6.5) x 10(6) per kg, respectively. The median number of transfused colony-forming units-granulocyte-macrophage was 12.45 (3.4-55.3) x 10(4) per kg. All patients showed rapid and sustained engraftment. The mean times to reach a neutrophil count of 0.5 x 10(9) per L and a platelet count of 50 x 10(9) per L were 11.50 +/- 1.70 and 13.90 +/- 3.98 days, respectively. All patients are alive and without transfusion requirements in complete remission 2 to 8 months after transplantation. CONCLUSION: This simplified cryopreservation technique will be useful for institutions without rate-controlled freezing facilities. Moreover, this method diminishes the amount of DMSO infused to patients, as well as its toxicity.

A simplified method for cryopreservation of hematopoietic stem cells with -80 degrees C mechanical freezer with dimethyl sulfoxide as the sole cryoprotectant.

A simplified method for cryopreservation was developed with 10% dimethylsulfoxide (DMSO) as the sole cryoprotectant without rate-controlled freezing. This method produced high recovery rate for mononucleated cells (87%) and elevated trypan blue viability (90%). Autologous peripheral blood stem cells (PBSCs) and bone marrow cells with plasma and 10% DMSO were frozen and stored in a -80 degrees C mechanical freezer. Eleven patients with solid and hematological malignancies were transplanted with autologous bone marrow or PBSCs. The median number of infused mononuclear cells (MNC) and CD34+ cells were 3.63 x 10(8)/Kg and 4.80 x 10(6)/Kg, respectively. The median number of infused post-thawing CFU-GM was 20 x 10(4)/Kg. All patients showed a rapid and sustained engraftment. The mean times to reach a neutrophil count of 0.5 x 10(9)/L and a platelet count of 50 x 10(9)/L were 11 and 13 days, respectively. All patients are alive and 10 in unmaintained complete remission for 3-9 months after transplantation. These results show the efficacy of this simplified cryopreservation technique that will be useful for institutions without rate-controlled freezing facilities.

Neonatal mixed lineage acute leukaemia.

We report here an uncommon case of neonatal acute leukaemia that presented concomitant with serological evidence of rubella infection. The clinical course was aggressive and the patient died 5 days after diagnosis from septicaemia. Leukaemic blasts had a mixed lineage immunophenotype co-expressing a constellation of B-lymphoid (CD19, cytCD22, TdT) and myeloid (CD13, CD33, CD14, anti-MPO) markers, as well as multiple adhesion molecules and markers associated with early lympho-myeloid progenitor cells (CD34, CD7, HLA-DR). A previously unrecorded discordant expression of different CD10 and CD34 epitopes was identified using different monoclonal antibodies. The karyotype was 46,XX t(4;11)(q21;q23) and molecular analysis confirmed rearrangement of the trithorax-related oncogene HRX at 11q23. There was a clonal biallelic rearrangement of the immunoglobulin heavy-chain gene. The features of this rare case have implications for possible aetiological events leading to leukaemia.

[Infective morbidity of replacement therapy in congenital coagulation deficiencies and its effects on demand of coagulation factors]. / La morbilidad infecciosa del tratamiento sustitutivo en los déficit congénitos de coagulación y su repercusión en la demanda de los factores de coagulación.

BACKGROUND: Analysis of the infective morbidity and mortality secondary to replacement hemotherapy in the population with congenital coagulation deficiencies (CCD) and their consequences on the demand for coagulation factors. METHODS: The 46 patients with CCD diagnosed in the autonomous community (AC) of the Balearic Islands (32 with hemophilia A, 6 with hemophilia B, 4 with von Willebrand's disease 2 with factor VII, 1 with factor X, and 1 with factor XII deficiencies) were investigated for infective morbidity and use of blood products from 1982 to 1987. RESULTS: 97% of the patients had some hepatitis marker, 77% had antibodies against human immunodeficiency virus (HIV) and 17% fulfilled the criteria for the acquired immunodeficiency syndrome (AIDS). There were 7 deaths (15%). The morbidity and mortality increased with age and use of blood products. There was a 46% reduction in factor VIII use between 1982 and 1986, from a mean yearly consumption per hemophiliac patient of 33444 international units (IU) to 18080 IU. CONCLUSIONS: The study results show a high prevalence of hepatitis and HIV, an important reduction in the demand of manufactured coagulation factors and a 15% reduction in the population with CCD during the study years.

Visceral leishmaniasis: another HIV-associated opportunistic infection? Report of eight cases and review of the literature.

Visceral leishmaniasis (VL) is considered an opportunistic infection in immunocompromised patients. We review the clinical, laboratory, and therapeutic data in 63 patients (eight new cases and 55 cases reported in the literature) with Mediterranean VL (kala azar) and HIV-1 infection to determine whether VL should be considered an opportunistic infection in HIV-infected adults. We conclude that: (1) in areas where both leishmaniasis and HIV-1 infection are endemic, VL may be more frequent among HIV-infected adults; (2) in HIV-infected patients, the clinical picture did not differ significantly from classical kala azar, although it often ran a recurrent course, with resistance to antimonial therapy. We propose the inclusion of VL in the IVC-2 subgroup of the Centers for Disease Control (CDC) clinical classification of HIV-1 infection while prospective and larger studies further define whether there are clinical presentations that could justify adding VL to the list of opportunistic infections indicative of AIDS.

Pre-B Acute Lymphoblastic Leukemia During a Complete Remission in T-Acute Lymphoblastic Leukemia.

This case provides evidence for a phenotypic switch from a characteristic T-ALL into a typical pre-B ALL. A 17 years old boy presented with massive hepatosplenomegaly and a mediastinal mass. The blasts were characterized as L2 type according to the French, American and British (FAB) classification and the staining with acid phosphatase was positive. The immunophenotype was CD2 +, CD5 +, CD7 + and CD10 -. The patient achieved a complete remission but relapsed 4 years later. At relapse, a striking immunological shift was apparent. The blast cells were now morphologically LI subtype and displayed the CD10 +. C{mew} + phenotype but were CD2 -, CD5 -, CD7 -. These findings are consistent with a transformation of the initial clone although the development of a second leukemia in the same patient could not be excluded with complete certainty.

Serum ferritin in haemodialysis.

A rapid assay for serum ferritin was performed in old and new patients undergoing regular dialysis treatment. The mean values were higher in the patients, especially the older patients, than in the controls. The normal differences in serum ferritin with age and sex were absent in the patients. We attribute these changes to iron loss and multiple transfusions received. Tissue iron was well reflected by serum ferritin concentration even during iron treatment, unless the intravenous route was used. Although an acceptable rise of serum ferritin was obtained, results of the iron administration have been poor.