Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:2.7.7.48 (transcriptase)
 9,479 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Peptides of melanosomal proteins have recently been shown to be recognized in an HLA-restricted mode by specific cytolytic T lymphocytes in melanoma patients. Dendritic antigen-presenting cells (DC) are considered to be the most effective stimulators of T cell responses, and the use of these cells has therefore been proposed to generate therapeutic responses to tumor antigens in cancer patients. We, therefore, generated DC from peripheral blood of normal donors in the presence of granulocyte/macrophage colony-stimulating factor and interleukin-4. Flow cytometric analysis of the cells during a 2-week culture revealed a loss of CD14 and CD34 expression, a concomittent increase of CD1a, CD11a,b and c, CD44, CD45, CD54, HLA-class I and II, and intermediate levels of CD26, CD80 and CD86. Cultured DC stimulated proliferation of allogeneic T cells and induced a marked, up to 20-fold, stimulation of T cell proliferation after pulsing with tetanus toxoid. To achieve independence of already-identified antigenic peptides presented in HLA class I-restricted fashion, which limits the general applicability of such peptides for vaccination of melanoma patients, we tested whether DC are transfectable with eukaryotic expression plasmids. DC transfected with two reporter genes (CAT, beta-galactosidase) using a liposome-based transfection technique, exhibited only low levels of enzymatically active proteins, but were able to degrade rapidly intracellular proteins and to process peptides efficiently. Chloramphenicol acetyltransferase as well as tyrosinase mRNA were detectable after transfection by reverse-transcriptase-polymerase chain reaction, and enzyme activities became measurable. Furthermore, DC transfected with the tyrosinase gene were able to induce specific T cell activation in vitro, indicating appropriate peptide processing and presentation in DC after transfection. These data suggest new approaches to future tumor vaccination strategies.
 ...
PMID:Dendritic cells generated from peripheral blood transfected with human tyrosinase induce specific T cell activation. 748 49

Expression of isoforms of estrogen receptor (ER) was examined in the bone tissues. Reverse transcriptase-polymerase chain reaction ((RT-PCR) using specific primers for rat ER cDNA was performed with total RNA from rat bone tissues. Then, we sequenced the amplified products after cloning and identified two isoforms of the ER and the wild-type ER. One of the ER mRNA isoforms did not have the region corresponding to exon 4 and the other isoform did not have the region corresponding to both exon 3 and exon 4. These isoforms were designated as ER delta 4 isoform and ER delta 3/4 isoform, respectively. The existence of these isoforms was also confirmed by ROS-17/2.8 osteoblastic osteosarcoma cells. Chloramphenicol acetyltransferase assay showed that these isoforms lost estrogen dependent transactivation activities. We suggest that the ER isoforms may play some roles in the bone metabolism in which estrogen is essential to maintain bone density.
 ...
PMID:Demonstration of isoforms of the estrogen receptor in the bone tissues and in osteoblastic cells. 858 81

Clinical and experimental studies showed that estrogen has antiatherogenic effects. We previously demonstrated that the estrogen receptor (ER) mRNA and protein are expressed in vascular smooth muscle cells (VSMC) derived from rat aorta. Here, the expression of isoforms of the ER was examined in VSMC. Reverse transcriptase-polymerase chain reaction using specific primers for rat ER cDNA was performed from RNA of rat VSMC. This revealed the existence of ER cDNA that is shorter than the wild-type ER cDNA. Sequencing of the amplified products identified three isoforms of the ER and the wild-type ER. These ER mRNA isoforms lacked the region corresponding to exon 4, exon 4 and 5, and exon 3 and 4. Therefore, they were designated as ERdelta4 isoform, ERdelta4/5 isoform and ERdelta3/4 isoform, respectively. Chloramphenicol acetyltransferase assay was performed with these ER isoforms constructed into the expression vector and the reporter plasmid containing the estrogen responsive element. The assay showed that these ER isoforms lost estrogen-dependent transactivation activities and that ERdelta4/5 isoform has a inhibitory effect on normal estrogen action when it was cotransfected with the wild-type ER. These ER isoforms might be involved in the regulation of VSMC by estrogen.
 ...
PMID:Identification of a novel isoform of estrogen receptor, a potential inhibitor of estrogen action, in vascular smooth muscle cells. 864 55

Desmoplastic small round cell tumor (DSRCT) is a unique, highly aggressive neoplasm that chiefly affects male adolescents and young adults. This tumor is characterized by nests of small undifferentiated cells that show immunohistochemical evidence of epithelial, mesenchymal, and neural differentiation. We report two cases of DSRCT that lacked immunohistochemical evidence of epithelial differentiation, but were found to have the fusion transcripts characteristic of this tumor. Both patients (a 41-year-old male and a 31-year-old female) presented with large intra-abdominal masses. After diagnostic biopsy, both were treated with multi-agent chemotherapy. One patient expired 18 days after diagnosis, and the other is currently alive 28 months later. Histologically, both tumors had the characteristic features of DSRCT and were composed of small round cells with hyperchromatic nuclei and scanty cytoplasm. In one of the cases, perinuclear intracytoplasmic hyaline inclusions were seen. Immunohistochemically, neither case expressed any of the epithelial markers tested, including AE1/AE3, CAM 5.2 and EMA. Both tumors were diffusely immunoreactive for desmin with a prominent globoid "dot-like" pattern of staining in one case. Both tumors stained for vimentin, neuron specific enolase, and synaptophysin, but were negative for CD99, muscle-specific actin, and myogenin. Reverse transcriptase-polymerase chain reaction revealed EWS-WT1 fusion transcripts characteristic of this neoplasm. In conclusion, we describe two cases of DSRCT that lacked immunohistochemical evidence of epithelial differentiation but had histologic and other immunohistochemical features which suggested this diagnosis. The ability to confirm the diagnosis of this rare tumor using molecular genetic techniques is particularly useful in those cases with unusual histologic or immunophenotypic features.
 ...
PMID:Cytokeratin-negative desmoplastic small round cell tumor: a report of two cases emphasizing the utility of reverse transcriptase-polymerase chain reaction. 1049 92