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1.
Clin Radiol ; 77(8): e628-e635, 2022 08.
Article in English | MEDLINE | ID: mdl-35688771

ABSTRACT

AIM: To assess the performance of a "triple-low" free-breathing protocol for computed tomography pulmonary angiography (CTPA) evaluated on patients with dyspnoea and suspected pulmonary embolism and discuss its application in routine clinical practice for the study of the pulmonary parenchyma and vasculature. MATERIAL AND METHODS: This study was conducted on a selected group of dyspnoeic patients referred for CTPA. The protocol was designed using fast free-breathing acquisition and a small, fixed volume (35 ml) of contrast agent in order to achieve a low-exposure dose. For each examination, radiodensity of the pulmonary trunk and ascending aorta, and the dose-length product (DLP) were recorded. A qualitative analysis was performed of pulmonary arterial enhancement and the pulmonary parenchyma. RESULTS: This study included 134 patients. Contrast enhancement of the pulmonary arteries (409 ± 159 HU) was systematically >250 HU. The duration of acquisition ranged from 0.9 to 1.3 seconds for free-breathing imaging. The mean DLP was in the range of low-dose chest CT acquisitions (145 ± 73 mGy·cm). The analysis was deemed optimal in 90% (120/134) of cases for the pulmonary parenchyma. Sixty-nine per cent (92/134) of cases demonstrated homogeneous enhancement of the pulmonary arteries to the subsegmental level. Only 6% (8/134) of examinations were considered uninterpretable. CONCLUSION: The present "triple-low" CTPA protocol allows convenient analysis of the pulmonary parenchyma and arteries without hindrance by respiratory motion artefacts in dyspnoeic patients.


Subject(s)
Pulmonary Embolism , Humans , Angiography/methods , Contrast Media , Dyspnea/diagnostic imaging , Pulmonary Artery/diagnostic imaging , Pulmonary Embolism/diagnostic imaging , Tomography, X-Ray Computed/methods
2.
Mutat Res ; 458(3-4): 55-70, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11691637

ABSTRACT

In many haematological diseases, and more particularly in B-cell chronic lymphocytic leukaemia (B-CLL), the existence of a tumour suppressor gene located within the frequently deleted region 13q14.3, has been put forward. A wide candidate region spanning from marker D13S273 to D13S25 has been proposed and an extensive physical map has been constructed by several teams. In this study, we sequenced a minimal core deleted region that we have previously defined and annotated it with flanking available public sequences. Our analysis shows that this region is gene-poor. Furthermore, our work allowed us to identify new alternative transcripts, spanning core regions, of the previously defined candidate genes DLEU1 and DLEU2. Since their putative involvement in B-CLL was controversial, our present study provide support for reconsidering the DLEU1 and DLEU2 genes as B-CLL candidate genes, with a new definition of their organisation and context.


Subject(s)
B-Lymphocytes/metabolism , Genes, Tumor Suppressor , Leukemia, Lymphocytic, Chronic, B-Cell/genetics , Sequence Deletion/genetics , Alternative Splicing/genetics , Base Sequence , Chromosome Mapping , Databases, Nucleic Acid , Exons/genetics , Expressed Sequence Tags , Humans , Molecular Sequence Data , Polymerase Chain Reaction , Repetitive Sequences, Nucleic Acid/genetics , Sequence Analysis, DNA
3.
Hum Genet ; 109(2): 178-85, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11511923

ABSTRACT

Slc30a4 is the fourth and last identified member of a mammalian proteins family presumably involved in the cellular transport of zinc, solute carrier family 30. The murine homologue of the human SLC30A4 gene has previously been investigated and found responsible for the lm, a phenotype due to zinc deficiency. According to the strong homology between mouse and human SLC30A4 coding sequences, and to the very similar clinical features encountered in the murine lm and in human acrodermatitis enteropathica, SLC30A4 has appeared to us to be a good candidate for acrodermatitis enteropathica. Here we detail the genomic structure of human SLC30A4 together with its localization on chromosome 15q15-q21. We also report the mutational analysis of human SLC30A4 in ten families with acrodermatitis enteropathica, which enabled us to exclude this gene from any involvement in the disorder of the patients examined.


Subject(s)
Acrodermatitis/genetics , Carrier Proteins/genetics , Chromosomes, Human, Pair 15/genetics , Acrodermatitis/metabolism , Amino Acid Sequence , Base Sequence , Blotting, Northern , Cation Transport Proteins , Chromosome Mapping , Chromosomes, Artificial/genetics , DNA Mutational Analysis , DNA Primers/chemistry , Genetic Linkage , Humans , In Situ Hybridization, Fluorescence , Molecular Sequence Data , Promoter Regions, Genetic , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction
4.
Hum Mutat ; 18(3): 212-24, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11524732

ABSTRACT

Using allele-specific amplification method (ARMS), a highly sensitive one-stage allele-specific PCR, we have evaluated the incidence of NRAS and KRAS2 activating mutations (codons 12, 13, and 61) in 62 patients with either monoclonal gammopathy of undetermined significance (MGUS) or multiple myeloma (MM), primary plasma-cell leukemia (P-PCL), and also in human myeloma cell lines (HMCL). NRAS and/or KRAS2 mutations were found in 54.5% of MM at diagnosis (but in 81% at the time of relapse), in 50% of P-PCL, and in 50% of 16 HMCL. In contrast, the occurrence of such mutations was very low in MGUS and indolent MM (12.50%). Of note, KRAS2 mutations were always more frequent than NRAS. The validity of the technique was assessed by direct sequencing of cell lines and of some patients. Multiple mutations found in two patients were confirmed by subcloning exon PCR amplification products, testing clones with our method, and sequencing them. Thus, these early mutations could play a major role in the oncogenesis of MM and P-PCL.


Subject(s)
Genes, ras/genetics , Leukemia, Plasma Cell/genetics , Multiple Myeloma/genetics , Alleles , Base Sequence , DNA Mutational Analysis , DNA, Neoplasm/chemistry , DNA, Neoplasm/genetics , Gene Frequency , Humans , Leukemia, Plasma Cell/diagnosis , Multiple Myeloma/diagnosis , Mutation , Polymerase Chain Reaction/methods , Tumor Cells, Cultured
5.
Leukemia ; 13(10): 1630-2, 1999 Oct.
Article in English | MEDLINE | ID: mdl-10516767

ABSTRACT

The chromosomal region 13q14.3 is frequently deleted in B cell chronic lymphocytic leukemia (B-CLL) and it is supposed that a tumor suppressor gene, involved in this leukemogenesis, is located in this area. The first exons of two genes, Leu1 and Leu2, mapped in a minimally deleted 13q14.3 region, are systematically lost in B-CLL sharing a 13q14.3 deletion. These two genes have been proposed as strong tumor suppressor gene candidates. However, in a study on 15 13q14.3 deleted B-CLL, we found three patients in which this critical region was not involved. Because of these results and that no mutations were detected on the two genes in a previous study, we think that Leu1 and Leu2 can be excluded as tumor suppressor genes.


Subject(s)
CD5 Antigens/genetics , CD8 Antigens/genetics , Chromosome Deletion , Chromosomes, Human, Pair 13 , Genes, Tumor Suppressor , Leukemia, Lymphocytic, Chronic, B-Cell/genetics , Humans , Loss of Heterozygosity , Polymerase Chain Reaction
6.
Genomics ; 54(1): 99-106, 1998 Nov 15.
Article in English | MEDLINE | ID: mdl-9806834

ABSTRACT

We report the characterization of a new gene mapped at chromosome band 13q14.3 telomeric to the retinoblastoma gene. This gene, designated CHC1L (for chromosome condensation 1-like), is composed of 14 exons spanning 30 kb of genomic DNA and encodes a ubiquitously expressed 3-kb mRNA. The N-terminal half of the deduced amino acid sequence shows strong homology with the seven tandem repeat structure of the regulator of chromosome condensation RCC1, which acts as a guanine nucleotide exchange factor (GEF) protein for the Ras-related GTPase Ran. CHC1L appears to be a new member of the RCC1-related GEF family.


Subject(s)
Cloning, Molecular , Proteins/genetics , Amino Acid Sequence , Base Sequence , Chromosome Mapping , Chromosomes, Human, Pair 13/genetics , DNA, Complementary , Guanine Nucleotide Exchange Factors , Humans , Molecular Sequence Data , Proteins/chemistry , Proteins/metabolism , RNA, Messenger/analysis , Reverse Transcriptase Polymerase Chain Reaction , Sequence Alignment , Tumor Cells, Cultured , ras Guanine Nucleotide Exchange Factors
7.
Genomics ; 52(3): 369-73, 1998 Sep 15.
Article in English | MEDLINE | ID: mdl-9790756

ABSTRACT

An expression map containing 48 ESTs was constructed to identify a tumor-suppressor gene involved in B-cell chronic lymphocytic leukemia (B-CLL), which was previously assigned to chromosome band 13q14.3 close to genetic markers D13S25 and D13S319. Thirty-nine of these 48 ESTs, together with 11 additional ones listed in databases, were initially assigned to chromosome 13q14 between markers D13S168 and D13S176. Nine others have recently been located in the D13S319 region. Our results indicate that 48 of the 59 ESTs analyzed belong to a YAC contig of chromosome 13 band q14, and 22 are contained on YAC 933e9, which encompasses the B-CLL critical region. Ten of these 22 ESTs were accurately assigned on a PAC, BAC, and cosmid contig encompassing the smallest minimal deletion area described so far in B-CLL, and 20 were tested for their expression on 27 normal or tumor tissues. One EST appears to be a likely candidate for the tumor-suppressor gene involved in B-CLL.


Subject(s)
Chromosomes, Human, Pair 13/genetics , Gene Expression Regulation, Neoplastic/genetics , Genes, Tumor Suppressor/genetics , Leukemia, Lymphocytic, Chronic, B-Cell/genetics , Chromosome Mapping , Chromosomes, Artificial, Yeast/genetics , Databases, Factual , Expressed Sequence Tags , Humans , RNA, Messenger/genetics , Sequence Deletion/genetics
8.
Genomics ; 46(2): 183-90, 1997 Dec 01.
Article in English | MEDLINE | ID: mdl-9417905

ABSTRACT

A putative tumor suppressor gene involved in B cell chronic lymphocytic leukemia (B-CLL) was mapped to human chromosome 13q14.3 close to the genetic markers D13S25 and D13S319. We constructed a 780-kb-long contig composed of cosmids, bacterial artificial chromosomes, and bacteriophage P1-derived artificial chromosomes that provides essential information and tools for the positional cloning of this gene. The conting contains both flanking markers as well as several additional genetic markers, three ESTs, and one potential CpG island. In addition, using one B-CLL patient, we characterized a small internal deleted region of 550 kb. Comparing this deletion with other recently published deletions narrows the minimally deleted area to less than 100 kb in our physical map. This deletion core region should contain all or part of the disrupted in B cell malignancies tumor suppressor gene.


Subject(s)
Chromosomes, Human, Pair 13 , Cosmids/genetics , Leukemia, Lymphocytic, Chronic, B-Cell/genetics , Sequence Deletion , Bacteriophages/genetics , Chromosomes, Artificial, Yeast , Chromosomes, Bacterial , Genetic Markers , Heterozygote , Humans , In Situ Hybridization, Fluorescence , Sequence Tagged Sites
9.
Clin Cancer Res ; 2(10): 1673-7, 1996 Oct.
Article in English | MEDLINE | ID: mdl-9816115

ABSTRACT

Fluorescence in situ hybridization with a chromosome 12-specific alpha-centromeric probe and a 13q14 yeast artificial chromosome probe was performed on interphase cells from 100 patients with B-cell chronic lymphocytic leukemia. Thirty-one patients exhibited a 13q14 deletion. No correlation was found between 13q14 deletions and clinical stage, sex, or morphology. Sixteen patients had trisomy 12, including 6 (of 12) with an atypical morphology. Trisomy 12 and 13q14 abnormalities were detected concomitantly in three patients only. The analysis of patients with deletions clearly showed that in five cases a significant number of cells retained two signals with the yeast artificial chromosome probe, indicating a genetic heterogeneity among the leukemic population. Our data confirm that the 13q14 deletion is a frequent event, indicate that the concomitant occurrence of 13q14 deletion and trisomy 12 is rare but possible, and show that both abnormalities are secondary events in B-cell chronic lymphocytic leukemia.


Subject(s)
Chromosome Deletion , Chromosomes, Human, Pair 13/genetics , Leukemia, Lymphocytic, Chronic, B-Cell/genetics , Chromosomes, Artificial, Yeast , Chromosomes, Human, Pair 12/genetics , DNA Probes , Genetic Markers , Humans , In Situ Hybridization, Fluorescence , Leukemia, Lymphocytic, Chronic, B-Cell/pathology , Trisomy
10.
Cancer Res ; 55(6): 1355-7, 1995 Mar 15.
Article in English | MEDLINE | ID: mdl-7882336

ABSTRACT

The presence of an unidentified tumor suppressor gene on the long arm of chromosome 13 which could be involved in the development of B cell chronic lymphocytic leukemia has been suspected because of frequent deletions of the locus D13S25 which lies 1.6 cM telomeric to the retinoblastoma gene. In order to accurately map this gene, cells from 25 B cell chronic lymphocytic leukemia tumors have been analyzed for allelic loss using a panel of microsatellite markers located in this region. These markers, which stretch from the retinoblastoma gene to the Wilson disease gene, have been ordered for their rank from centromere to telomere. In addition to the data obtained from deletion pattern of these markers, results from preliminary pulse-field electrophoresis studies enable us to redefine the minimal deleted area from more than 1 cM to 280 kilobase around D13S25.


Subject(s)
Chromosome Deletion , Chromosomes, Human, Pair 13 , Genes, Tumor Suppressor , Leukemia, Lymphocytic, Chronic, B-Cell/genetics , Chromosome Mapping , Humans
11.
Hum Genet ; 94(4): 450-1, 1994 Oct.
Article in English | MEDLINE | ID: mdl-7927349

ABSTRACT

We describe seven restriction fragment length polymorphisms (RFLPs) in the Leukemia Inhibitory Factor (LIF) gene region. These new markers, found using a cosmid contig, have been used to map precisely the chromosome 22 long arm.


Subject(s)
Genetic Markers , Growth Inhibitors/genetics , Interleukin-6 , Lymphokines/genetics , Polymorphism, Genetic , Alleles , Chromosome Mapping , Chromosomes, Human, Pair 22 , Cloning, Molecular , DNA/analysis , Genetic Linkage , Humans , Leukemia Inhibitory Factor , Polymorphism, Restriction Fragment Length , Repetitive Sequences, Nucleic Acid
13.
J Clin Invest ; 91(1): 12-20, 1993 Jan.
Article in English | MEDLINE | ID: mdl-8423212

ABSTRACT

The mechanisms responsible for skin lesions during acute graft-vs.-host disease (aGVHD) after allogeneic bone marrow transplantation (BMT) are poorly understood. The exact role of various effector cell populations and "major" (particularly HLA-DP) or "minor" antigens as target molecules is not known. To investigate the nature of cells responsible for tissue injury, we cultured T cells from skin biopsy first with interleukin 2 (IL-2) alone and then in polyclonal activation conditions to avoid in vitro antigenic sensitization before specificity testing. We applied this method to two biopsies performed during aGVHD after semiallogeneic BMT and obtained cytotoxic T cells against four graft mismatches: CD8+ T cells against HLA-A2.2 and HLA-B27 and CD4+ T cells against HLA-DP101 and HLA-DP401. This demonstrates that T cells with documented specificity can be obtained from an aGVHD lesion without antigenic selection. Moreover, these data directly implicate DP as a potential target antigen for aGVHD.


Subject(s)
Bone Marrow Transplantation/immunology , Graft vs Host Disease/immunology , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/surgery , Leukemia, Myeloid, Acute/surgery , Skin/immunology , T-Lymphocytes/immunology , Adult , Antigens, CD/analysis , Blotting, Southern , Female , Gene Rearrangement, T-Lymphocyte , HLA-A Antigens/analysis , HLA-B Antigens/analysis , HLA-DP Antigens/analysis , HLA-DQ Antigens/analysis , HLA-DR Antigens/analysis , Histocompatibility Testing , Humans , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/immunology , Leukemia, Myeloid, Acute/immunology , Lymphocyte Activation , Male , Skin/pathology , T-Lymphocyte Subsets/immunology , Transplantation, Homologous
14.
Hum Immunol ; 29(2): 110-6, 1990 Oct.
Article in English | MEDLINE | ID: mdl-2249950

ABSTRACT

A panel of 34 clones was established from a cell line derived from the skin biopsy of a patient (genotype: A1, A2, B7, B8, DR3, DR6) undergoing acute graft-versus-host disease after semiallogeneic bone marrow transplantation with his mother's bone marrow (genotype: A1, A1, B7, B8, DR3, DR6). The T-cell line obtained presented the following phenotype: CD3+, CD4+, CD8-, CD16-, WT31+, T-cell receptor delta 1-, 4B4+, 2H4-, CD25+, DR+. This CD4+ T-cell line was poorly cytotoxic against the target cells tested, including the mother's phytohemagglutinin blasts as a negative control (autologous T cells), the father's phytohemagglutinin blasts bearing the mismatch haplotype, K562, U937, SVK14 (a keratinocyte cell line), and a panel of B-lymphoblastoid cell lines bearing HLA-A2, the known mismatch antigen. All but 1 of the 34 clones obtained were of CD4+ phenotype, and none was CD16+. Only the sole CD8+ clone showed significant cytotoxicity against the father's phytohemagglutinin blast; however, this cytotoxic activity was associated with the highest score for nonspecific killing against both K562 and U937. This work demonstrates the feasibility of obtaining a large panel of clones from a graft-versus-host disease target organ to constitute the basic cellular material for in vitro study of the graft-versus-host process.


Subject(s)
Graft vs Host Disease/pathology , Skin/pathology , T-Lymphocyte Subsets , T-Lymphocytes/pathology , Acute Disease , Bone Marrow Transplantation , Cells, Cultured , Child , Clone Cells/pathology , Cytotoxicity Tests, Immunologic , Humans , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/pathology , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/surgery , Male
15.
Transpl Int ; 3(3): 176-80, 1990 Oct.
Article in English | MEDLINE | ID: mdl-2271090

ABSTRACT

The immune repertoire of T lymphocytes invading human allografts is of fundamental importance both at the operational level, in order to achieve relevant matching, and at the functional level, since the unique capacity of T and B cells to specifically recognize allogeneic components restricts the origin of the signals leading to rejection by these cells. In this paper, the authors review their own work, as well as other contributions in this domain, with special reference to the frequency and function of donor-committed cells among the infiltrate and the relationship between T-cell receptor gene rearrangements and repertoire.


Subject(s)
T-Lymphocytes/immunology , Transplantation Immunology , Clone Cells/immunology , Humans , Lymphokines/biosynthesis , Phenotype
16.
Blood ; 74(1): 285-90, 1989 Jul.
Article in English | MEDLINE | ID: mdl-2546620

ABSTRACT

Repeated analysis of peripheral blood lymphocytes (PBLs) from a patient with large granular lymphocytosis, neutropenia, and rheumatoid arthritis revealed that approximately 45% of PBLs displayed the following phenotype: CD3+, CD4-, CD8-, CD16+, HNK-1-, WT31-. This population was purified for further analysis by depletion with anti-CD4 and anti-CD8 monoclonal antibodies (MoAbs). Southern blot analysis showed preferential rearrangements of the V gamma 9 genes. Northern blot demonstrated the presence of V gamma 9 mRNA transcripts. With MoAbs directed against either the V gamma 9 peptide (Ti gamma A) or the delta chain of the gamma delta T-cell receptor (TCR delta 1), we further demonstrated that those cell surfaces expressed both V gamma 9 and a delta gene product. In addition, analysis of the gamma gene rearrangements on six clones derived from this population demonstrated a unique rearrangement on a single chromosome, strongly suggesting the monoclonality of this T-cell population. Significant cytotoxic activity against K562, U937 was observed only after an in vitro culture period with interleukin-2 (IL-2), whereas no specific inhibitory effect on autologous bone marrow (BM) CFU-G was noted.


Subject(s)
Agranulocytosis/physiopathology , Arthritis, Rheumatoid/physiopathology , Lymphocytes/physiology , Neutropenia/physiopathology , Receptors, Antigen, T-Cell/physiology , Antigens, Differentiation/analysis , Bone Marrow Cells , Cytotoxicity, Immunologic , Female , Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor , Hematopoietic Stem Cells/physiology , Humans , Immunity, Cellular , Lymphocytes/classification , Middle Aged , Receptors, Antigen, T-Cell, gamma-delta
17.
Virology ; 151(2): 390-3, 1986 Jun.
Article in English | MEDLINE | ID: mdl-3010560

ABSTRACT

The replicative form (RF) of spiroplasma virus 4 (SpV4) has been purified from infected cells of Spiroplasma melliferum strain G1 by alkaline lysis followed by low melting point agarose gel electrophoresis. A partial restriction map has been established. The circular RF was linearized by cutting at the unique ClaI restriction site and has been cloned in Escherichia coli HB101 using the plasmid pBR328 as a vector. The recombinant plasmid was purified by equilibrium centrifugation in ethidium bromide-cesium chloride gradient. After ClaI endonuclease digestion, the inserted SpV4 RF DNA was recovered by low melting point agarose gel electrophoresis and was recircularized by ligation. The cloned SpV4 RF DNA was demonstrated to be infectious by transfection.


Subject(s)
Bacteriophages/genetics , Cloning, Molecular , Spiroplasma/genetics , Transfection , DNA Restriction Enzymes , Escherichia coli/genetics
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