Pérez Sánchez C, Ayensa F, Lloveras E, Zamora L, Cirigliano V, Pérez E, Plaja A
Prenatal diagnosis of an interstitial 12q chromosome deletion.
Ann. Genet. , 47(2)177-9.
Rearrangements involving long arm of chromosome 12 are rare events. To our knowledge, we present the first case of an interstitial deletion of the long arm of chromosome 12 in a prenatal diagnosis. A review of the literature is included in our report.
Complete characterization of the 3' region of the human and mouse hereditary hemochromatosis HFE gene and detection of novel splicing forms.
Blood Cells Mol. Dis. , 27(1)35-43.
The human HFE gene was identified in 1996 as the gene whose mutations are responsible for hereditary hemochromatosis in most patients. Expression analysis by Northern blot indicated that the gene was approximately 4.1 kb in length. However, the cDNA reported was only 2716 bp. These results implied that at least 1.4 kb of the mRNA remained to be identified. In the present study, we detected several 3' EST clones while screening the genomic region of the gene in search of potential additional HFE mRNA sequences. Subsequent sequencing of these EST clones and RT-PCR experiments revealed that exon 7 of the HFE gene has, in fact, a length of 1944 bp and it presents two polyadenylation signals. The new human HFE exon 7 region has been screened in non-C282Y HH patients in search for new putative mutations. Mouse 3' RACE experiments also further extend the previously reported mouse HFE exon 6 sequence. Additionally, we report two novel end forms of the human HFE gene detected by 3' RACE experiments and several novel splicing forms identified in the HepG2 cell line.
Kudo A, Yamamoto F, Furusawa M, Kuroiwa A, Natori S, Obinata M
Structure of thymidine kinase gene introduced into mouse Ltk- cells by a new injection method.
Gene , 19(1)11-9.
Pricking, a new injection method developed by Yamamoto et al. (1981), can be used to introduce DNA into cultured cells with high efficiency. Closed circular plasmid DNA containing the cloned HSV-TK gene (pTK-1) was introduced by this method and the structure of DNA in stable transformants was examined. In most clones, the introduced DNA was integrated into the mouse genome in a tandemly repeated form. The possibility of multiple integration via mouse middle repetitive sequences was also examined using the chimeric plasmid with TK genes and middle repetitive sequences (pMRTK-1). Digestion with restriction enzymes showed that the middle repetitive sequence used in this experiment had no effect on the efficiency of transformation, suggesting that this sequence is unable to mediate homologous recombination with mouse genomes.
Kahn S, Yamamoto F, Almoguera C, Winter E, Forrester K, Jordano J, Perucho M
The c-K-ras gene and human cancer (review).
Anticancer Res. , 7(4A)639-52.
A significant number of human tumors from diverse histological origins contain c-K-ras oncogenes which have been activated by somatic point mutations resulting in single amino acids substitutions in the encoded p21 ras protein. In addition to these qualitative changes, other genetic alterations leading to increased expression of the c-K-ras gene, especially its mutated form, appear to be important in the activation of its oncogenic potential. These findings support the hypothesis that c-K-ras oncogenes are contributing in a dominant but dose dependent manner to the multistage process of human tumorigenesis. Activated c-K-ras oncogenes have been detected in human tumors at different stages of progression, including premalignant neoplasms. These studies provide evidence for the involvement of somatic mutational activation of this ras gene in the early stages of tumor development in some types of human cancer. We discuss here the structural and functional features of the human c-K-ras gene and the involvement of its activated form in human cancer.
Human Embryonic and Induced Pluripotent Stem Cells
Chapter in the book: Human Embryonic and Induced Pluripotent Stem Cells
Chapter title: Induced pluripotent stem cells from cord blood CD133+ cells using Oct4 and Sox2
Published by Springer Protocol Handbook Series in 2011
Díaz-Ricart M, Carretero M, Castillo R, Ordinas A, Escolar G
Digital image analysis of platelet-extracellular matrix interactions: studies in von Willebrand disease patients and aspirin-treated donors.
Haemostasis , 24(4)219-29.
We have developed an automated image analysis method which provides en face morphometric evaluation of platelet interactions with coverslips coated with extracellular matrices. This procedure gives also additional information on the intensity of platelet-surface interactions in the same specimens. To standardize the method, anticoagulated human blood was flowed at 800 s-1 during 10 min at 37 degrees C through a parallel perfusion chamber in which extracellular matrix (ECM)-coated coverslips were placed. The sensitivity of the new method was tested in similar studies performed in patients with von Willebrand disease or treated with aspirin (500 mg/day). The extent of platelet deposition on the ECMs was evaluated by both standard and new morphometric procedures. Coverslips were successively embedded in resin and processed for cross-sectional analysis and results were compared with those provided by the new automated procedure. A good correlation was found between results of covered surface obtained by both methods (r = 0.86; p < 0.005). Results from perfusions performed with blood from patients with platelet defects confirmed the correspondence of grey levels in the digitized images with the heights of groups of interacting platelets calculated from cross sections. The new automated method, in only one stage, allows a reliable evaluation of the global interaction of platelets with the ECM and effectively detected qualitative modifications in platelet-platelet interactions in patients with known platelet defects. The method described could be applied to the evaluation of the interactions of platelets with purified components of the subendothelium.
Sierra J, Grañena A, Bosch F, Carreras E, Martí JM, Urbano-Ispizua A, Rovira M, Rozman C
Mitoxantrone and intermediate-dose cytosine arabinoside for poor-risk acute leukemias: response to treatment and factors influencing outcome.
Hematol Oncol , 10(6)301-9.
Mitoxantrone (MIT, 12 mg/m2, i.v. 5 days) and intermediate-dose cytosine arabinoside (IDAC 1 g/m2/12 h, i.v. 3 days) was given to 43 patients with poor-risk acute leukemias (AL). Moderate or severe toxicity was infrequent. The proportion of complete remissions (CR) in the main patient categories was as follows: 15/18 (85 per cent) in acute myeloid leukemia (AML) in the first relapse, 2/6 in ALL in the first relapse, 0/2 in AML in relapse after bone marrow transplantation (BMT), 2/7 in AML refractory to first-line treatment (REF-AL), and 1/6 in postmyelodysplastic (PMD-AL) plus secondary AL (S-AL). The mortality rate during induction was 23 per cent. Median duration of CR was 24 weeks. The multivariate prognostic factor analysis on CR obtention showed that data concerning treatment for the first relapse and platelet count higher than the median of the series were favourable. On the contrary, PMD-AL, S-AL and REF-AL were unfavourable situations. A percentage of marrow erythroblasts superior to the median was a favourable prognostic factor for survival. Finally, the duration of CR after MIT-IDAC was directly related to the duration of previous CR. In conclusion, MIT-IDAC was highly effective to attain CR in AML in the first relapse. However, due to the poor long-term results in these patients, additional measures are recommended after CR.
Blanco Y, Saiz A, Costa M, Torres-Peraza JF, Carreras E, Alberch J, Jaraquemada D, Graus F
Evolution of brain-derived neurotrophic factor levels after autologous hematopietic stem cell transplantation in multiple sclerosis.
Neurosci. Lett. , 380(1-2)122-6. Epub 1 Feb 2005
A neuroprotective role of inflammation has been suggested based on that immune cells are the main source of brain-derived neurotrophic factor (BDNF). We investigated the 3-year evolution of BDNF levels in serum, CSF and culture supernatant of peripheral blood mononuclear cells (PBMC), unstimulated and stimulated with anti-CD3 and soluble anti-CD28 antibodies, in 14 multiple sclerosis patients who underwent an autologous hematopoietic stem cell transplantation (AHSCT). BDNF levels were correlated with previously reported MRI measures that showed a reduction of T2 lesion load and increased brain atrophy, mainly at first year post-transplant. A significant decrease of serum BDNF levels was seen at 12 months post-transplant. BDNF values were found significantly lower in stimulated but not in unstimulated PBMC supernatants during the follow-up, supporting that AHSCT may induce a down-regulation of BDNF production. The only significant correlation was found between CSF BDNF levels and T2 lesion load before and 1 year after AHSCT, suggesting that BDNF reflects the past and ongoing inflammatory activity and demyelination of these highly active patients. Our study suggests that AHSCT can reduce BDNF levels to values associated with lower activity. This decrease does not seem to correlate with the brain atrophy measures observed in the MRI.
Urbano-Ispizua A, García-Conde J, Brunet S, Hernández F, Sanz G, Petit J, Bargay J, Figuera A, Rovira M, Solano C, Ojeda E, de la Rubia J, Rozman C
High incidence of chronic graft versus host disease after allogeneic peripheral blood progenitor cell transplantation. The Spanish Group of Allo-PBPCT.
Haematologica , 82(6)683-9.
The incidence of acute GVHD (aGVHD) in allogeneic peripheral blood progenitor cell transplantation (allo-PBPCT) seems to be similar to that seen in allogeneic bone marrow transplantation (allo-BMT). In contrast, some preliminary results suggest that the incidence of chronic GVHD (cGVHD) might be higher. The aim of the present study was to analyze the actuarial probability of developing cGVHD in allo-PBPCT, its clinical manifestations and response to treatment.