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J Coll

Publications and source records attributed to J Coll.

At least 37 records · Page 2Linked to original sources

Isolation and characterization of a chicken homologue of the Spi-1/PU.1 transcription factor.

Spi-1/PU.1 is a member of the Ets family of transcription factors important in regulation of hematopoiesis. We have isolated a chicken cDNA homologuous to the mammalian Spi-1/PU.1 gene with an open reading frame of 250 amino acids (aa). The chicken Spi-1/PU.1 protein is 14 aa and 16 aa shorter than its human and mouse counterparts but is extremely well conserved with 78.8% and 75.2% identity respectively. The carboxy terminal DNA binding region, or ETS binding domain, is 100% identical to that of human and mouse. Some differences with the mammalian homologues are seen in the N-terminal part of the protein and in the PEST connecting domain. However, the differences are mainly conservative and all the features underlying functional aspects seem preserved. The major discrepancy lies in a 12 aa deletion in an already poorly conserved part of the PEST sequence. Spi-1/PU.1 transcripts were detected at high levels in spleen and Fabricius bursa of chick embryos by Northern blot and in situ hybridization. Our results show that the chicken Spi-1/PU.1 protein behaves like a bonafide Spi-1/PU.1 transcription factor in its DNA binding and transactivating properties.

Amino Acid Sequence↗

H19 overexpression in breast adenocarcinoma stromal cells is associated with tumor values and steroid receptor status but independent of p53 and Ki-67 expression.

In a previous study we described the expression of the H19 gene by in situ hybridization (ISH) in normal breast and in benign or malignant breast tumors (Dugimont T, Curgy JJ, Wernert N, Delobelle A, Raes MB, Joubel A, Stehelin D, Coll J: Biol Cell 1995, 85:117-124). In the present work, 1) we extend the previous one to a statistically useful number of adenocarcinomas, including 10 subclasses, 2) we provide information on the precise ISH localization of the H19 RNA by using, on serial tissue sections, antibodies delineating specifically the stromal or the epithelial component of the breast, and 3) we consider relationships between the H19 gene expression and various clinicopathological information as tumor values (T0 to T4), grades, steroid receptors, lymph node status, and molecular features as the p53 gene product and the Ki-67/MIB1 protein, which is specific to proliferating cells. Data indicate that 1) in 72.5% of studied breast adenocarcinomas an overall H19 gene expression is increased when compared with healthy tissues, 2) the H19 gene is generally overexpressed in stromal cells (92.2%) and rarely in epithelial cells (2.9% only), 3) an up-regulation of the H19 gene is significantly correlated with the tumor values and the presence of both estrogen and progesterone receptors, and 4) at the cellular level, the H19 gene demonstrates an independent expression versus accumulation of both the p53 protein and the Ki-67/MIB-1 cell-cycle marker.

Adenocarcinoma↗

A new indole derivative from the red alga Chondria atropurpurea. Isolation, structure determination, and anthelmintic activity.

Chondriamide C (3), a new bis(indole) amide, was isolated from the red alga Chondria atropurpurea, and its structure was established from spectroscopic data and chemical transformations. A new natural product, 3-indoleacrylamide (4), and the previously described chondriamides A and B (1, 2) and 3-indoleacrylic acid (5) were also isolated. The anthelmintic activities of compounds 1, 3, 4, and 6 (the O,N1,N1'-trimethyl derivative of compound 2) against Nippostrongylus brasiliensis in vitro were evaluated.

Albendazole↗

Lacrimal immunoglobulins in rheumatoid arthritis patients with or without Sjögren's syndrome.

The aim of the present study was to analyse immunoglobulin G, A, and M levels in tears of patients with rheumatoid arthritis with or without keratoconjunctivitis sicca (KS) which define Sjögren's syndrome (SS). Tears were collected from the lower cul-de-sac by capillarity (100-300 microl). Tear IgG, IgA and IgM levels were determined by radial immunodiffusion. Samples diluted 1:10 were used for IgA and IgG and non-diluted ones for IgM. Fifty-three patients with rheumatoid arthritis and 30 healthy control subjects were studied. In all individuals IgA predominated in tears, IgG levels were low but with a very wide range and IgM was present in very low concentrations. The IgG concentration in tears showed statistically significant differences between the control group and that of rheumatoid arthritis and KS. IgG in tears correlated positively with the rose bengal test (r=0.2848, p<0.05) and negatively with the Schirmer test (r=0.3042, p<0.05). Tear IgG measurement might provide a marker for eye involvement in patients with rheumatoid arthritis and KS which define SS.

Adult↗

Isolation and characterization of transformed human T-cell lines infected by Epstein-Barr virus.

Epstein-Barr virus (EBV) is a human lymphotropic virus whose main targets have traditionally been described as B lymphocytes and epithelial cells. Here we report the isolation and characterization of largely monoclonal transformed human T-cell lines infected by EBV. The transformed T cells expressed CD2, CD3, and either CD4 or CD8 surface molecules and more generally displayed the phenotype of naive T cells with a complete and clonal rearrangement of the T-cell receptor. None of the cell lines expressed B cells, natural killer, or myeloid antigens or had immunoglobulins genes rearrangement. They grew in the absence of growth factor; however, they all secreted interleukin-2 after mitogenic activation. Polymerase chain reaction (PCR) analysis showed the presence of EBV DNA in all these cell lines. Moreover, Southern blot analysis of one of these cell lines shows the presence of circular episomic EBV DNA, and by Northern blot or reverse transcriptase-PCR analysis, only the expression of Epstein-Barr nuclear antigen-1 (EBNA-1) and latent membrane protein-1 (LMP-1) genes was detected. Finally, the complete transformed phenotype of this T-cell line was shown by its injection into nude or recombination activating gene 2 (RAG2)-deficient mice that led to the formation of solid tumors.

Animals↗

[Aphthae].

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Acquired Immunodeficiency Syndrome↗

Neutralization susceptibility of African swine fever virus is dependent on the phospholipid composition of viral particles.

In this study we have investigated the generation of African swine fever (ASF) virus variants resistant to neutralizing antibodies after cell culture propagation. All highly passaged ASF viruses analyzed were resistant to neutralization by antisera from convalescent pigs or antibodies generated against individual viral proteins which neutralized low-passage viruses. A molecular analysis of neutralizable and nonneutralizable virus isolates by sequencing of the genes encoding for neutralizing proteins revealed that the absence of neutralization of high-passage viruses is not due to antigenic variability of critical epitopes. A comparative analysis of phospholipid composition of viral membranes between low- and high-passage viruses revealed differences in the relative amount of phosphatidylinositol in these two groups of viruses, independent of the cells in which the viruses were grown. Further purification of low- and high-passage viruses by Percoll sedimentation showed differences in the phospholipid composition identical to those found with the partially purified viruses and confirmed the susceptibility of these viruses to neutralization. The incorporation of phosphatidylinositol into membranes of high-passage viruses rendered a similar neutralization susceptibility to low-passage viruses, in which this is a major phospholipid. In contrast, other phospholipids did not interfere with high-passage virus neutralization, suggesting that phosphatidylinositol is essential for a correct epitope presentation to neutralizing antibodies. Additionally, the removal of phosphatidylinositol form a low-passage virus by a specific lipase transformed this virus from neutralizable to nonneutralizable. These data constitute clear evidence of the importance of the lipid composition of the viral membranes for the protein recognition by antibodies and may account in part for the past difficulties in reproducibly demonstrating ASF virus-neutralizing antibodies by using high-passage viruses.

African Swine Fever Virus↗

Soluble interleukin-2 receptor in primary and secondary Sjögren's syndrome.

Serum levels of soluble interleukin-2 receptor (sIL2-R) were studied in patients with or without Sjögren's syndrome (SS). The mean sIL2-R level was significantly higher (P < 0.0001) in those with autoimmune diseases, whether with or without SS, than in healthy people. The serum level of sIL2-R did not help to distinguish primary from secondary SS, or the absence of this syndrome in patients with autoimmune diseases. We found a correlation between the sIL2-R level and the presence of SS-A and SS-B antibodies or antinuclear antibody levels, and no correlation with other serological and clinical markers. Salivary gland anomalies were always accompanied by high sIL2-R levels, but there was no correlation between sIL2-R levels and degree of glandular lesion. Routine determination of sIL2-R in patients with dry syndrome is likely to prove useful for ruling out SS without resorting to invasive testing.

Antibodies, Antinuclear↗

Immunohistochemistry of minor salivary gland biopsy specimens from patients with Sjögren's syndrome with and without hepatitis C virus infection.

OBJECTIVES: To characterise phenotypically the minor salivary glands of patients with clinical and histological features of Sjögren's syndrome (SS) infected with hepatitis C virus (HCV). PATIENTS AND METHODS: 75 consecutive patients with SS (31 primary SS, 44 secondary SS) diagnosed by preliminary European classification criteria. The presence of anti-HCV antibodies was detected by commercial third generation ELISA and by a second generation immunoblot assay. Presence of HCV genome in serum was determined by polymerase chain reaction analysis. Expression of CD3, CD4, CD8, CD20, HLA-DR, and CD25 molecules in lymphocytic and epithelial cells on minor salivary glands was detected by immunohistochemical assays. Expression of interferon gamma and interleukin 4 cytokines was determined by in situ hybridisation. RESULTS: Six of 31 primary SS (19%) and one of 44 secondary SS (2%) serum samples were positive for anti-HCV by ELISA. Three samples were positive, three indeterminate, and one sample corresponding to a secondary SS patient was negative by immunoblot. The three immunoblot positive serum samples were also HCV-RNA positive by PCR assay. The study of lymphocytic cells in the diffuse infiltrate of minor salivary glands showed a predominance of the CD3 lymphocytic population. A predominance of CD4 over CD8 T cells (ratio 2:1) was observed in HCV and non-HCV infected patients. The analysis of the lymphocytic focus showed that the HCV infected patients had a predominance of CD20 positive cells. Activation molecules (CD-25 and HLA-DR) were expressed in HCV and non-HCV infected patients in lymphocytic and epithelial cells, however epithelial cell expression of CD25 was low in HCV infected patients. As expected, a pronounced Th1 response was observed in the lymphocytic foci of HCV patients. CONCLUSIONS: HCV infected patients may develop an autoimmune sialadenitis, similar to that described in primary SS.

Adult↗

High prevalence of subclinical Sjögren's syndrome features in patients with autoimmune thyroid disease.

OBJECTIVE: To determine the prevalence of keratoconjunctivitis sicca and xerostomia related to Sjögren's syndrome (SS) in asymptomatic patients with diagnosed autoimmune thyroid diseases (AITD); and to investigate whether the immunopathologies of sialadenitis observed in AITD associated SS and primary SS are similar. METHODS: One hundred seventy-six patients diagnosed with AITD (88 with Graves' disease, 40 Hashimoto's thyroiditis, 48 primary myxedema) were tested for keratoconjunctivitis sicca (Schirmer's test and rose bengal staining) and for xerostomia (salivary scintigraphy and labial salivary gland biopsy). Immunohistopathological studies were performed on cryostat sections of bucal mucosa biopsies using antibodies to CD3, CD4, CD8, CD20, CD14, CD25, LFA-1, ICAM-3 HLA class II, tumor necrosis factor-alpha, interleukin 1, and interferon-gamma. RESULTS: Nineteen of 52 (37%) patients with AITD fulfilled the criteria for xerostomia and 39/170 (23%) for keratoconjunctivitis sicca. Features of SS were diagnosed in 43 of 176 (24%) patients with AITD, with similar prevalence in Graves' (20%). Hashimoto's thyroiditis (27%), and primary myxedema (29%). In AITD associated SS, infiltrating lymphocytes were mainly CD3+ T lymphocytes, with a CD4/CD8 ratio of 2:1. In most patients infiltrating lymphocytes expressed activation markers, HLA class II molecules, and interleukin 2 receptor (CD25). In some patients HLA class II was inappropriately expressed in the epithelial gland cells. CONCLUSION: The finding that a third of patients with AITD have SS features confirms that AITD and SS are mutually associated. Together with the similarity of immunopathology of sialadenitis in AITD associated SS in primary SS, this supports the theory that SS and AITD are 2 autoimmune diseases closely related pathogenetically.

Adolescent↗

High incidence of loss of heterozygosity and abnormal imprinting of H19 and IGF2 genes in invasive cervical carcinomas. Uncoupling of H19 and IGF2 expression and biallelic hypomethylation of H19.

The few imprinted genes characterized so far include the insulin-like growth factor-2 gene (IGF2) coding for a foetal growth factor and the H19 gene whose normal function is unknown but which is likely to act as an RNA with an antitumour effect. IGF2 is expressed by the paternal allele and H19 by the maternal allele. This reciprocal expression is quite interesting because both H19 and IGF2 genes are located close to each other on chromosome 11p15.5 in a region subject to loss of heterozygosity (LOH). Moreover, loss of imprinting (LOI) or biallelic expression has been proposed as an epigenetic mechanism for tumorigenesis in a variety of human cancers including Wilms' tumour. In this study we report the LOH, LOI and methylation status of H19 and IGF2 genes in 29 invasive cervical carcinomas of different clinical stages. Fourteen (48%) and 13 (45%) tumours were heterozygous for H19 and IGF2 respectively. LOH for H19 and IGF2 genes were found in 2 of 14 (14%) and 3 of 13 (23%) informative tumours, respectively. LOI of H19 and IGF2 was detected in 2 of 12 (17%) and 5 of 10 (50%) tumours with no LOH, respectively. More interestingly, monoallelic expression of the otherwise silent H19 allele (allele switch) was observed in 2 of 12 (17%) tumours and biallelic expression of IGF2 was detected in one specimen of normal cervix adjacent to the tumour. The expressing H19 allele, and to a lower degree also the silent allele, were hypomethylated in tumours suggesting that demethylation of both H19 alleles may be associated with an early step of imprinting alteration. In cervical cancer H19 and IGF2 expressions could be independently regulated. In conclusion, our data suggest that H19 and IGF2 genes, via deletions and/or abnormal imprinting, could play a crucial role in a large proportion (58%) of cervical cancers where they may be associated with disease progression.

Alleles↗

Increased radionuclide uptake on bone scintiscans: a common but not clinically significant finding for human immunodeficiency virus type 1-infected patients free of osteoarticular symptoms.

The aim of this study was to describe findings of bone scintiscans and their clinical significance for patients infected with human immunodeficiency virus type 1 (HIV-1); 33 HIV-1-infected patients (22 men and 11 women) free of osteoarticular symptoms were included in the study. Plain bone roentgenograms, bone mineral contents (measured by dual-photon absorptiometry), and scintiscans (determined with 99mTc diphosphonate) were obtained for all subjects. Plain bone roentgenograms showed no abnormalities, and bone mineral contents were within the normal range for all patients. Radionuclide bone scans were unremarkable for eight patients (24%) and showed symmetrical abnormally increased uptake in the epiphyseal region of the appendicular skeleton in 25 (76%). Follow-up of the patients for > or = 1 year ruled out subsequent development of osteoarticular disorders. Generalized, symmetrical increased radionuclide uptake on bone scans is a common finding for HIV-1 infected patients free of osteoarticular symptoms. This finding is probably related to bone marrow hypercellularity and is of no clinical significance; therefore, other diagnostic procedures are not required in the workup of these patients.

Adolescent↗

Assessment of the European classification criteria for Sjögren's syndrome in a series of clinically defined cases: results of a prospective multicentre study. The European Study Group on Diagnostic Criteria for Sjögren's Syndrome.

OBJECTIVE: To assess the recently proposed preliminary criteria for the classification of Sjögren's syndrome (SS) in a multicentre European study of a new series of clinically defined cases. METHODS: The criteria included six items: I = ocular symptoms; II = oral symptoms; III = evidence of keratoconjunctivitis sicca; IV = focal sialoadenitis by minor salivary gland biopsy; V = instrumental evidence of salivary gland involvement; VI = presence of autoantibodies. Each centre was asked to provide five patients with primary SS, five with secondary SS, five with connective tissue diseases (CTD) but without SS, and five controls (patients with ocular or oral features that may simulate SS). The preliminary six item classification criteria set was applied to both the SS patients and the non-SS controls, and the performance of the criteria in terms of sensitivity and specificity was tested. RESULTS: The criteria set was tested on a total of 278 cases (157 SS patients and 121 non-SS controls) collected from 16 centres in 10 countries. At least four of the six items in the criteria set (limiting item VI to the presence of Ro(SS-A) or La(SS-B) antibodies) were present in 79 of 81 patients initially classified as having primary SS (sensitivity 97.5%), but in only seven of 121 non-SS controls (specificity 94.2%). When the presence of item I or II plus any two of items III-V of the criteria set was considered as indicative of secondary SS, 97.3% (71 of 73) of the patients initially defined as having this disorder and 91.8% (45 of 49) of the control patients with CTD without SS were correctly classified. CONCLUSION: This prospective study confirmed the high validity and reliability of the classification criteria for SS recently proposed by the European Community Study Group.

Adult↗

The 5' part of the human H19 RNA contains cis-acting elements hampering its translatability.

H19 is an imprinted gene developmentally regulated in man and mouse and implicated in various neoplasms. No corresponding protein product has yet been detected, although several open reading frames (ORFs) could be identified along its RNA. The largest ORF found in the human gene could encode a putative 26 kDa protein. We have isolated two H19 cDNAs (AP and ES) that contain this ORF4 and correspond to incomplete copies of the unique 2.3 kb H19 RNA. In transient expression assays, AP was able to synthesize a 26 kDa protein whereas ES was not. With respect to ORF4, ES exhibits a 536 bp long GC-rich 5' untranslated region, whereas AP contains the last 22 nucleotides of this 5'UTR. Using deletions and point mutations, we have found that the length and probably the secondary structure of the 5'UTR strongly hampers the translatability of the RNA. In addition, a potential role of upstream ORFs (uORFs) was detected as stressed by the enhances translation of a construct mutated in uORF3 overlapping ORF4. Interactions between H19 and proteins are indicated by a specific binding between 5'UTR derived RNA segments and two nuclear proteins of about 27 kDa. Our results favor a potential role of these particular structures and binding properties in general trans-regulation of RNA post-transcriptional processes rather than in normal control of H19 mRNA translation.

Animals↗