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Biomedical subjects

C Jin

Publications and source records attributed to C Jin.

At least 109 records · Page 6Linked to original sources

Sequence specific collective motions in a winged helix DNA binding domain detected by 15N relaxation NMR.

The recognition between transcription factors and their DNA binding sites is a highly dynamic process. During transcriptional regulation, transcription factors must bind to or dissociate from their cognate DNA binding sites. The winged helix DNA binding motif is one of many highly conserved DNA binding motifs identified in transcription factors. Backbone dynamics has been studied on the 15N- and 2H-enriched winged helix family member Genesis. Our data show that the overall motions of the single domain Genesis are better described by more than two autocorrelation times (taum). Our data also demonstrate that Genesis shows structure specific conformation exchange characterized by Rex. Therefore, our results indicate that the structure of Genesis is highly dynamic and that secondary structure elements in Genesis have collective motions in the nanosecond to millisecond time scale. Since the winged helix DNA binding motif is highly conserved, this unique dynamic property observed in Genesis is also likely to be conserved in other winged helix family members and important in DNA binding.

Animals↗

Characterization of a malignant eccrine poroma by cytogenetic and fluorescence in situ hybridization techniques.

We have cytogenetically and by fluorescence in situ hybridization (FISH) analyzed short-term cultures from a malignant eccrine poroma, a type of tumor in which chromosomal abnormalities have previously not been reported. Combining the results from G-banding and FISH, the karyotype was interpreted as 50,XX,t(3;11;12)(q21;p15;q15),+5,+7,+9, +9,t(12;22)(q13;q13),+del(12)(q15),der(15)t(3;15) (q23;p13),der(17)t(8;17)(q22;p13),-20,add(22)(q13).

Acrospiroma↗

Unbalanced chromosomal rearrangements in a metastasizing salivary gland tumor with benign histology.

Benign metastasizing pleomorphic adenoma (BMPA) is a rare tumor of the salivary glands. Despite benign histopathologic features, it can metastasize and is sometimes lethal. No chromosomal data have been reported for this tumor type. We have by chromosome banding and fluorescence in situ hybridization analysis examined the short-term cultures of three skeletal metastases from a BMPA and identified two related hypodiploid clones: 44,XX,dic(3;22)(p11;q13) or der(3)t(3;22)(p11;q?) add(22)(q?),der(9;21)(q10;q10),der(13)t(1;13)(q11;p13)/45,XX,-3,der(9;21 ) (q10;q10),der(13)t(1;13)(q11; p13),?der(22)t(3;22)(q22;q13), +mar. The karyotypic features of this BMPA thus differ from the characteristic cytogenetic findings in pleomorphic adenomas and carcinomas ex pleomorphic adenoma.

Adenoma, Pleomorphic↗

Radioresistant cell strain of human fibrosarcoma cells obtained after long-term exposure to x-rays.

A radioresistant cell strain from human fibrosarcoma HT 1080 has been obtained after prolonged exposure to x-rays for 7 months (2 Gy per day, 5 days per week). This new strain, HT1080R, differs from HT1080 in a significantly increased ability of clonogenical survival, with coefficient alpha decreasing from 0.161 to 0.123 Gy(-1) and coefficient beta decreasing from 0.0950 to 0.0565 Gy(-2). Furthermore, the radioresistance of HT1080R proved to be stable in long-term passaged cultures as well as in frozen samples. Differences between the two cell lines are also observed in the G-banded karyotype; the new cell line shows monosomy of chromosome 17 and loss of 5p+ and 11q+ present in the parental cells. These data suggest that the radioresistance may have been caused by radiation-induced cell mutation and that the resistant cells may have been selected by repeated irradiations. In order to characterize this new strain, the ability of the cells to rejoin DNA double-strand breaks, the cell cycle distribution and the amount of apoptosis after irradiation have been estimated; however, no differences are observed between these two cell strains. Although the mechanism of the elevated radioresistance remains unknown, this pair of cell strains can provide a new model system for further investigations with regard to the mechanisms of cellular radioresistance. The results also show that any type of irradiation similar to the schedules used in radiotherapy can lead to the formation and selection of more radioresistant cell clones in vitro, a phenomenon with possible implications for radiotherapy.

Apoptosis↗

Nonrandom numerical chromosome abnormalities in basal cell carcinomas.

Clonal chromosome abnormalities were found in 22 of 23 short-term cultured basal cell carcinomas (BCC) of the skin. The karyotypic abnormalities were nonrandom and in several cases included evidence of clonal evolution. Especially in cultures showing an epithelial growth pattern, simple numerical changes, most commonly +18, +9, +20, +7, and +5, predominated and presumably constitute pathogenetically important aberrations present in the neoplastic parenchyma. Also, several structural rearrangements of chromosome arm 9q were seen, which may be of particular interest against the background that a gene for familial BCC (Gorlin syndrome), the PTCH gene, maps to this region. Finally, most of the clonal aberrations detected in predominantly fibroblast-like cultures are likely to reflect changes acquired by cells of the tumor stroma, which raises the question whether mutations also of this tumor component may play a pathogenetic role in BCC development.

Adult↗

Cytogenetic and molecular genetic demonstration of polyclonality in an acinic cell carcinoma.

The paradigm that human malignancies are monoclonal has been questioned during recent years by the finding of unrelated, cytogenetically aberrant clones in short-term cultures from certain tumour types, notably carcinomas of the breast, skin and upper aerodigestive tract. In order to analyse whether cytogenetically unrelated clones are also unrelated at the molecular level, we analysed the X-chromosome inactivation status in cell cultures from a cytogenetically highly polyclonal acinic cell carcinoma of the parotid gland. By using cell cultures dominated by a single abnormal clone, obtained through in vitro culturing for 3-5 passages, we showed that the different clones must indeed have originated from different cells.

Aged↗

Screening for diabetic retinopathy using a non-mydriatic retinal camera in rural Victoria.

PURPOSE: To develop a screening programme for the early detection of diabetic retinopathy using non-mydriatic retinal photography. METHODS: A community based screening service was offered to all people with known diabetes mellitus in selected townships in the LaTrobe and Goulburn Valleys in Victoria. At the local examination centre, basic sociodemographic information was collected as well as details of previous use of eye care services for the early detection of diabetic retinopathy. The examination included visual acuity (VA), glycosylated haemoglobin level and Polaroid photographs of each fundus using a Canon CR5-45NM non-mydriatic retinal camera (Canon, Tochigiken, Japan). Dilating drops were not used. Photographs were subsequently reviewed and letters were sent to all participants (with copies to their general practitioners) with recommendations for appropriate follow up. RESULTS: A total of 1177 people with diabetes attended the screening service, which is estimated to be 40% of the total population with known diabetes in the study area. The mean age was 65 years (range 20-94 years); 559 (48%) people reported not having a dilated fundus examination within the past 2 years; 345 (29%) people had never had a dilated fundus examination. Of the 2354 eyes, 2126 (90%) of the photographs were gradable. A total of 704 people (60%) had normal VA and no evidence of diabetic retinopathy, 209 people (18%) had diabetic retinopathy, 101 people (9%) had evidence of other fundus pathology, 42 people (3%) had reduced acuity (< 6/18) in one or both eyes (with no fundus pathology evident) and 121 people (10%) had ungradable photographs in one or both eyes. CONCLUSIONS: The present study demonstrates the usefulness of a screening programme with non-mydriatic retinal photography as an adjunct to current eye care services for the early detection of diabetic retinopathy.

Adult↗

Dissociative identity disorder in psychiatric inpatients.

OBJECTIVE: The aim of this study was to replicate reports of a high rate of dissociative identity disorder in psychiatric inpatients. METHOD: Subjects were 100 randomly selected women, 16-50 years old, who had recently been admitted to an acute psychiatric hospital. Diagnoses were made by two interviewers through use of the Structured Clinical Interview for DSM-IV Dissociative Disorders. RESULTS: One percent (N = 1) of the interviewed subjects had dissociative identity disorder. CONCLUSIONS: Contrary to previous studies, the authors found a low rate of dissociative identity disorder, perhaps because of the different methodology used.

Adolescent↗

[Clinical observation on artificial shexiang baoxin pill in treating 112 patients of coronary heart disease with angina pectoris].

OBJECTIVE: To test and verify the effect of artificial Shexiang Baoxin Pill (SXBXP, a heart protecting musk pill). METHODS: Two hundred and twenty-five patients of coronary heart disease (CHD) with angina pectoris were randomly and blindly assigned to the study group (112 cases) and the control group (113 cases). The two groups were well matched in age, sex, angina type, complication and grade of cardiac function. Two pills of artificial or natural SXBXP were given to each cases of the two groups for 3 times every day orally, for 2 weeks, respectively. RESULTS: Both the artificial and natural SXBXP could not only reduce significantly the frequency of angina episode and the daily consumption of nitroglycerin, they also could result in remarkable improvement of NST, sigma ST and exercise tolerance, as well as significant reduction of rate-pressure product. The total symptomatic effective rate of the two groups was 86.6% and 89.4% respectively, and the ECG effective rate, 52.7% and 57.5% respectively (P > 0.05), and the incidence of headache of the two groups was one for eoch. In addition, the onset of action of both agents was rapid, 88.5% of the study group and 78.6% of the control group were within five minutes. The sustained time of action of the two groups was also similar, i.e. 4.9 +/- 4.4 h vs 5.4 +/- 3.1 h. CONCLUSION: The clinical efficacy and side effect of artificial SXBXP were very similar to those of natural SXBXP.

Adult↗

FISH characterization of head and neck carcinomas reveals that amplification of band 11q13 is associated with deletion of distal 11q.

In order to characterize homogeneously staining regions (HSR) and other 11q13 rearrangements identified cytogenetically, we performed fluorescence in situ hybridization (FISH) using a CCND1 cosmid and five YAC clones spanning chromosomal bands 11q13-14 on metaphase cells from 14 primary and one metastatic head and neck carcinomas. At the cytogenetic level, a total of 17 HSR were detected in ten cases: five were in derivative chromosomes 11 in band 11q13, and 12 were located in other derivative chromosomes. Other forms of 11q13 rearrangements were observed in five cases, whereas two cases had normal chromosomes 11. FISH analysis demonstrated that all HSR but two were derived from the 11q13 band. The size of the amplicon varied from case to case, but the amplification always included the region covered by YAC 55G7, which contains the CCND1 locus. The amplification of CCND1 was confirmed by use of a CCND1 cosmid. We also showed that most of the cases (9 of 11) with 11q13 amplification had lost material from distal 11q. The breakpoints were mapped by FISH and were shown to cluster to the region between YACs 55G7 and 749G2. We conclude that loss of gene(s) in distal 11q may be as important as amplification of genes in 11q13 for the biological aggressiveness of head and neck carcinomas.

Adult↗

Evidence that the DNA binding specificity of winged helix proteins is mediated by a structural change in the amino acid sequence adjacent to the principal DNA binding helix.

We present the first structural evidence supporting the hypothesis that the binding specificity of the winged helix DNA binding motif is mediated by residues adjacent to the alpha-helix (H3), the moiety which is primarily involved in the interaction with DNA. Using NMR to determine secondary structural elements of a winged helix family member, Genesis (formerly HFH-2), and comparing these with those found in the X-ray crystal structure of the HNF-3gamma/DNA complex [Clark, K. L., Halay, E. D., Lai, E., & Burley, S. K. (1993) Nature 364, 412-420], we show that the major differences observed occur for H3 and the region immediately prior to this. H3 in Genesis is slightly shorter than in HNF-3gamma and, in addition, we observe an extra small helix (H4) in the region between H2 and H3 which is not found in the HNF-3gamma/DNA complex. This is significant as it has been shown previously [Overdier, D. G., Porcella, A., & Costa R. H. (1994) Mol. Cell. Biol. 14, 2755-2766] that the DNA-binding specificity is influenced by amino acid residues in this region.

Amino Acid Sequence↗

COX15 codes for a mitochondrial protein essential for the assembly of yeast cytochrome oxidase.

The respiratory defect of Saccharomyces cerevisiae mutants assigned to complementation group G4 of a pet strain collection stems from their failure to synthesize cytochrome oxidase. The mutations do not affect expression of either the mitochondrially or nuclearly encoded subunits of the enzyme. The cytochrome oxidase deficiency also does not appear to be related to mitochondrial copper metabolism or heme a biosynthesis. These data suggest that the mutants are likely to be impaired in assembly of the enzyme. A gene designated COX15 has been cloned by transformation of mutants from complementation group G4. This gene is identical to reading frame YER141w on chromosome 5. To facilitate further studies, Cox15p has been expressed as a biotinylated protein. Biotinylated Cox15p fully restores cytochrome oxidase in cox15 mutants, indicating that the carboxyl-terminal sequence with biotin does not affect its function. Cox15p is a constituent of the mitochondrial inner membrane and, because of its resistance to proteolysis, probably is largely embedded in the phospholipid bilayer of the membrane. The present studies further emphasize the complexity of cytochrome oxidase assembly and report a new constituent of mitochondria involved in this process. The existence of COX15 homologs in Schizosaccharomyces pombe and Caenorhabditis elegans suggests that it may be widely distributed in eucaryotic organisms.

Amino Acid Sequence↗

SHY1, the yeast homolog of the mammalian SURF-1 gene, encodes a mitochondrial protein required for respiration.

C173 and W125 are pet mutants of Saccharomyces cerevisiae, partially deficient in cytochrome oxidase but with elevated concentrations of cytochrome c. Assays of electron transport chain enzymes indicate that the mutations exert different effects on the terminal respiratory pathway, including an inefficient transfer of electrons between the bc1 and the cytochrome oxidase complexes. A cloned gene capable of restoring respiration in C173/U1 and W125 is identical to reading frame YGR112w of yeast chromosome VII (GenBank Z72897Z72897). The encoded protein is homologous to the product of the mammalian SURF-1 gene. In view of the homology, the yeast gene has been designated SHY1 (Surf Homolog of Yeast). An antibody against the carboxyl-terminal half of Shy1p has been used to localize the protein in the inner mitochondrial membrane. Deletion of part of SHY1 produces a phenotype similar to that of G91 mutants. Disruption of SHY1 at a BamHI site, located approximately 2/3 of the way into the gene, has no obvious phenotypic consequence. This evidence, together with the ability of a carboxyl-terminal coding sequence starting from the BamHI site to complement a shy1 mutant, suggests that the Shy1p contains two domains that can be separately expressed to form a functional protein.

Cloning, Molecular↗

Cytogenetic analysis of inverted nasal papillomas and demonstration of genetic convergence during in vitro passaging.

Three inverted nasal papillomas were cytogenetically investigated after short-term culture. Two of the cases were characterized by a single abnormal clone with t(1;8)(p36;q11) and trisomy 7, respectively, whereas the third papilloma showed extreme cytogenetic heterogeneity: of 852 analyzed cells, 329 belonged to 36 unrelated clones, 344 had non-clonal changes, and 179 had a normal chromosome constitution. The polyclonal papilloma was further analyzed during in vitro passage of 3 lines (L1-L3) cultured independently since initiation of the primary cultures and found to have 6, 16 and 6 unrelated clones at analysis of primary cultures. At passage 1, each line was further subdivided into 2 sub-lines (L1A and B, L2A and B, and L3A and B), which were cultured separately until the cells spontaneously stopped dividing. After 4 to 7 passages, each sub-line was dominated (83-98% of the cells) by a single clone. The cell populations that took over the cultures were the same within each set of sub-lines (A and B lines), demonstrating that clonal overgrowth in vitro is not random. The difference in clonal selection among the L1-L3 lines further shows that genetic convergence during in vitro growth in stable conditions is dependent not only on the clones' ability to adapt to the culture conditions, but also on the nature of the neighboring cells with which they collaborate and compete.

Aged↗

Clonal chromosome aberrations accumulate with age in upper aerodigestive tract mucosa.

Short-term cultured non-neoplastic upper aerodigestive tract (UAT) mucosa samples from 36 patients with squamous cell carcinoma of the head and neck (SCC) and 53 patients with benign UAT disorders were cytogenetically analyzed. The cell cultures were divided into two series: in series A, cells were cultured in a medium stimulating outgrowth of mesenchymal cells; whereas the cultured cells in series B were of epithelial morphology. Series A was further subdivided into three different age groups (< or = 15 years, 16-59 years, and > or = 60 years) of non-SCC patients and one SCC group. Series B was composed of two groups; one with and one without SCC. Among the non-SCC patients in series A, there was an increase with age in the frequency of cells/sample with numerical and structural chromosomal changes as well as in the incidence of clonal chromosomal aberrations. No differences could, however, be detected between cancer patients and age-matched controls. In series B, the frequency of cells/sample with numerical changes and the incidence of clonal numerical aberrations were significantly higher among SCC patients. Three main conclusions could be drawn. First, the frequencies of clonal and non-clonal chromosome aberrations in UAT mucosa were age dependent. Second, the cytogenetic support for the validity of the field cancerization hypothesis was restricted to increased levels of numerical chromosome changes in epithelial cell cultures from cancer patients. Third, clonal chromosome aberrations, including autosomal and sex chromosome aneuploidies as well as structural rearrangements, are not restricted to neoplastic mucosal cells.

Adolescent↗

The reciprocal translocation t(9;16)(q22;p13) is a primary chromosome abnormality in basal cell carcinomas.

The reciprocal translocation t(9;16)(q22;p13) was identified in three short-term cultured basal cell carcinomas (BCCs). The t(9;16) was the sole anomaly in one clone in two tumors and was accompanied by a second change that also affected the long arm of chromosome 9 in the third. In addition, other cytogenetically unrelated abnormal clones were also found in all three BCCs. The identification of t(9;16)(q22;p13) as a primary chromosomal abnormality in a subset of BCCs (we found it in 3 of 22 tumors) is especially intriguing against the background that the PTCH gene, which when mutated in the germ line presumably gives rise to the autosomal dominant basal cell nevus or Gorlin's syndrome, maps to chromosome band 9q22. None of the genes rearranged in the BCC-specific t(9;16)(q22;p13) translocation have been identified, but we hypothesize that the translocation represents the cytogenetic corollary of a tumorigenic recombination of PTCH with an as yet unknown gene in 16p13. If so, this would be the first time that a tumor suppressor gene causally involved in a hereditary cancer is shown to be frequently rearranged through a specific translocation in sporadic carcinomas of the same type.

Aged↗

Cloning and characterization of MRP10, a yeast gene coding for a mitochondrial ribosomal protein.

The nuclear gene MRP10 of Saccharomyces cerevisiae was cloned by complementation of a respiratory deficient mutant N518/L1. This mutant is defective in mitochondrial translation and shows a tendency to accumulate deletions in mitochondrial DNA (rho-). Analysis revealed Mrp10p to be a component of the 37 S subunit of the mitochondrial ribosomes. Disruption of MRP10 in a haploid strain of yeast elicits a phenotype identical to that of the original mutant. The respiratory defect of the null mutant is rescued by re-introducing the MRP10 gene in a wild-type mitochondrial DNA background. These results indicate that Mrp10p belongs to the class of yeast mitochondrial ribosomal proteins that are essential for translation. Searches of current databases failed to reveal any homologs among known bacterial or eucaryotic cytoplasmic ribosomal proteins. Some sequence similarity, however, was detected between Mrp10p and Yml37p, previously identified as a component of the yeast mitochondrial 50 S ribosomal subunit.

Amino Acid Sequence↗