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Biomedical subjects

C Y Yu

Publications and source records attributed to C Y Yu.

At least 37 records · Page 2Linked to original sources

The CACC box upstream of human embryonic epsilon globin gene binds Sp1 and is a functional promoter element in vitro and in vivo.

DNA sequences 179 base pairs upstream and 23 base pairs downstream of the cap site of human embryonic epsilon globin gene exhibit promoter activity in transfected cell cultures. The nuclear factor binding in vitro of this epsilon promoter region was studied by DNase I foot-printing, methylation interference, and gel mobility shift assay. Four major nuclear factor-binding sites are detected in complexes formed with unfractionated nuclear extracts: NF-E1 at -163, epsilon F1 at -143, CACC box at -111, and CBF at -81, respectively. Of these, NF-E1 is an erythroid-specific factor. epsilon F1 is probably a ubiquitous factor because it is present in both erythroid K562 cells and nonerythroid HeLa cells. The epsilon F1-binding site exhibits sequence similarity to that of the cAMP response element-binding protein family of transcription factors. Finally, the CCAAT box-binding protein (CBF)-binding site centers around the CCAAT promoter box. Comparative binding studies with unfractionated nuclear extracts and affinity purified HeLa Sp1 demonstrated that the epsilon-globin CACC box at -111 is a binding site of Sp1. The spatial arrangements of the NF-E1-binding site, the CACC box, and CCAAT box, with respect to their mutual separations by approximately integral numbers of helical turns, is well conserved in all mammalian embryonic epsilon globin promoters. Transient expression assay, using human growth hormone gene as the receptor, demonstrated that similar to the other two human beta-like globin genes (beta and gamma), the CACC box of epsilon globin gene is an essential promoter element for epsilon globin gene expression in vivo in K562 cells. This CACC promoter box also functions in vitro in nuclear extracts prepared from K562 cells. These data, together with Sp1-binding studies of the human beta and gamma globin CACC boxes, suggest that the general transcription factor Sp1, through its differential interactions with different forms of CACC promoter boxes, is an essential component of the machinery that controls the developmental program of mammalian globin gene regulation.

Base Sequence

The complete exon-intron structure of a human complement component C4A gene. DNA sequences, polymorphism, and linkage to the 21-hydroxylase gene.

The human complement component C4A and C4B genes are located within the class III region of the MHC. The polymorphic C4 genes are highly complex including variations in class (isotype), size, and number of genes. The DNA sequence for a C4A gene has been determined, except for a large intron of 6 to 7 kb long. The C4A gene consists of 41 exons encoding a transcript for a precursor protein of 1744 amino acid residues. Several structural and functional aspects of C4 have been located to individual exons. The active site of the anaphylatoxin C4a matches to a splice junction. Some unique properties of C4, such as, the alpha-gamma-chain junction, the tyrosine sulfation sites, and the post-secretory metalloprotease cleavage site, are encoded by a single exon. Comparison of human C4 with published data for mouse C4, human C3 and rat alpha 2 macroglobulin genes revealed that these evolutionary-related genes share very similar exon-intron structures. Altogether 20 polymorphic sites in human C4 have been detected by various techniques. Presumably, these polymorphic residues account for the functional, structural, and serologic variations observed among the various allotypes. A PvuII restriction length polymorphism has been detected within the region of DNA coding for C4a. The intergenic region between C4 and the neighboring 21-hydroxylase gene, CYP21, is approximately 3028 bp in size.

Base Sequence

Postirradiation angiosarcoma of the terminal ileum.

Sarcomas which develop after radiotherapy mainly involve the soft tissue and the bone. Postirradiation angiosarcomas involving the internal organs are exceedingly rare and so far only three cases have been reported. Here, a case of angiosarcoma of the terminal ileum in a 48-year-old female is reported. The tumor developed 39 months after radiotherapy for recurrent squamous cell carcinoma of the uterine cervix. Angiosarcoma recurred locally and metastasized to the liver eight months after initial resection of the primary lesion of the terminal ileum. The patient died from sepsis 23 days after the resection of the recurrent ileal and metastatic hepatic neoplasms. Hence, any patients complaining of new discomfort that occurs in an irradiated field two years or more after radiotherapy should be promptly searched for the presence of the postirradiation neoplasms.

Female

[Quantification of acetabular coverage in normal adult].

Quantification of acetabular coverage is important and can be expressed by superimposition of cartilage tracings on the maximum cross-sectional area of the femoral head. A practical Autolisp program on PC AutoCAD has been developed by us to quantify the acetabular coverage through numerical expression of the images of computed tomography. Thirty adults (60 hips) with normal center-edge angle and acetabular index in plain X ray were randomly selected for serial drops. These slices were prepared with a fixed coordination and in continuous sections of 5 mm in thickness. The contours of the cartilage of each section were digitized into a PC computer and processed by AutoCAD programs to quantify and characterize the acetabular coverage of normal and dysplastic adult hips. We found that a total coverage ratio of greater than 80%, an anterior coverage ratio of greater than 75% and a posterior coverage ratio of greater than 80% can be categorized in a normal group. Polar edge distance is a good indicator for the evaluation of preoperative and postoperative coverage conditions. For standardization and evaluation of acetabular coverage, the most suitable parameters are the total coverage ratio, anterior coverage ratio, posterior coverage ratio and polar edge distance. However, medial coverage and lateral coverage ratios are indispensable in cases of dysplastic hip because variations between them are so great that acetabuloplasty may be impossible. This program can also be used to classify precisely the type of dysplastic hip.

Acetabulum

Mosaic variegated aneuploidy with microcephaly: a new human mitotic mutant?

We report the case of a 17 year old girl with severe microcephaly and mental retardation, in whom karyotype analysis of PHA-stimulated lymphocytes, cultured skin fibroblasts, direct and cultured bone marrow and EBV-transformed lymphoblasts all showed at least 10% of cells with trisomy, which could be for many different chromosomes. All trisomies except 5, 10, 13, 14 and 17 were observed. Tissue-specific differences in the predominant trisomy occurred. The existence of this mosaic trisomy in four different tissues and in repeated cultures over a three year period suggests that it is due to a genetic abnormality resulting in mitotic instability. This case is compared with six previously reported human cases with a similar phenomenon, including two pairs of siblings. It is unclear whether all cases represent the same condition, since clinical and cytogenetic differences exist among them. The term "mosaic variegated aneuploidy with microcephaly" is suggested as a descriptive term for this syndrome.

Adolescent

Characterization of two hybrid C4 allotypes (C4A*12 and C4B*3) by electrophoretic, serological and restriction fragment length polymorphism analyses.

Informative pedigree analysis of two rare C4 allotypes is reported. One proband was C4A deficient as a consequence of having one haplotype with a deleted C4A gene, and the second haplotype with two C4B genes--one encoding the common C4B*1 and one encoding a unique hybrid gene product C4B*3. C4B*3 had approximately normal C4B hemolytic activity, a single alpha-chain of MR 94,000 by SDS-PAGE but was positive for Rg:1,2 by hemagglutination inhibition (HAI) and for Rg:1 by Western blotting. The hybrid nature was confirmed by RFLP analysis with a Rg:1-associated fragment by Eco0109 digestion but no C4A-associated fragments by N1aIV digestion were identified. A gene conversion at Locus I which included just the C4 isotype region could explain the structure of C4B*3. The second pedigree had a Rodgers negative C4A*12 allotype. This C4A gene, which segregated with a single 7.0 kb TaqI fragment, encoded a C4A alpha-chain, which was negative for Rg:1 epitope. The affected haplotype lacked the Rg:1-associated fragment by Eco0109 digestion yet had the C4A specific N1aIV digestion fragment. These studies successfully employed RFLP analyses to confirm serologic and electrophoretic observations.

Adolescent

Cell type-specific protein-DNA interactions in the human zeta-globin upstream promoter region: displacement of Sp1 by the erythroid cell-specific factor NF-E1.

The protein-DNA interactions of the upstream promoter region of the human embryonic zeta-globin gene in nuclear extracts of erythroid K562 cells and nonerythroid HeLa cells were analyzed by DNase I footprinting, gel mobility shift assay, methylation interference, and oligonucleotide competition experiments. There are mainly two clusters of nuclear factor-binding sites in the zeta promoter. The proximal cluster spans the DNA sequence from -110 to -60 and consists of binding sites for CP2, Sp1, and NF-E1. NF-E1 binding is K562 specific, whereas CP2 binding is common to both types of cells. Overlapping the NF-E1- and CP2-binding sites is a hidden Sp1-binding site or CAC box, as demonstrated by binding studies of affinity-purified Sp1. In the distal promoter region at -250 to -220, another NF-E1-binding site overlaps a CAC box or Sp1-binding site. Extract-mixing experiments demonstrated that the higher affinity of NF-E1 binding excluded the binding of Sp1 in the K562 extract. NF-E1 factors could also displace prebound Sp1 molecules. Between the two clusters of multiple-factor-binding sites are sequences recognized by other factors, including zeta-globin factors 1 and 2, that are present in both HeLa and K562 extracts. We discuss the cell type-specific, competitive binding of multiple nuclear factors in terms of functional implications in transcriptional regulation of the zeta-globin gene.

Base Sequence

Inflammatory pseudotumor of the liver with occlusive endophlebitis: report of a case.

Inflammatory pseudotumor (IPT) of the liver is an extremely rare benign tumor. Only 12 cases have so far been reported in the English literature. The etiology and pathogenesis of IPT remain obscure. We herein present an additional case of IPT of the liver with occlusive endophlebitis in a 36-year-old female. Immunohistochemical studies demonstrated the polyclonal nature of the plasma cells in the tumor, including IgA, IgM, IgG, kappa and lambda. The inflammatory pseudotumor should be kept in mind in the differential diagnosis of hepatic space-occupying malignant lesions.

Adult

[Effect of electroshock on learning-dependent long-term potentiation in rat hippocampal CA3 area].

Population spikes were recorded from hippocampal CA3 of freely moving rats following stimulation of the perforant path. Six rats were trained to perform conditioned drinking behavior. (1) Animals were treated with electroshock in less than 15 minutes after daily training session for a period of 6 consecutive days. Synaptic efficacy did not increase and the conditioned response could not be established during the behavioral training in four rats, synaptic efficacy was increased, the long-term potentiation (LTP) appeared and the conditioned behavior was established in other two rats. (2) After consolidation of conditioned behavior, electroshock temporarily caused suppression of the learning-dependent LTP and the correlative conditioned response. Both LTP and the correlative conditioned response recovered in one to four hours after electroshock. In these cases, the maximal development of LTP preceded the development of conditioned behavior. The results indicated that the learning-dependent LTP might be one of the neural substrates underlying learning and memory.

Animals

A physical map linking the five CD1 human thymocyte differentiation antigen genes.

Human CD1 is a family of thymocyte differentiation antigens which consist of heavy chains with mol. wts between 43 and 49 kd binding to beta 2 microglobulin. They are distant relatives of the major histocompatibility complex (MHC) class I and II products. Five human CD1 genes have been described. Three (CD1A, -B and -C) code for the serologically defined CD1a, -b and -c antigens. The protein products of the other two genes, CD1D and CD1E, remain unknown. All CD1 genes are located on chromosome 1 and hence are independent of the MHC locus. In this paper, the tight linkage of the CD1 genes has been established by pulse field gel electrophoresis, cosmid cloning and walking techniques. The 190 kb of DNA linking all five CD1 genes has been spanned by 14 overlapping cosmids. The order of the genes in the CD1 complex is CD1D-CD1A-CD1C-CD1B-CD1E, and, with the exception of CD1B, they are arranged in the same transcriptional orientation. The genes are evenly spaced in the complex except for the distance between CD1D and CD1A, which is two to three times greater than the average.

Antigens, CD

Unique sequence organization and erythroid cell-specific nuclear factor-binding of mammalian theta 1 globin promoters.

The theta 1 globin gene is an alpha globin-like gene, and started to diverge from the other members of the alpha globin family 260 million years ago. DNA sequencing and transcriptional analysis indicated that it is functional in erythroid cells of the higher primates, but not in prosimians and rabbit. The theta 1 promoter region of higher primates including man consists of GC-rich sequences characteristic of housekeeping gene promoters, and CCAAT and TATA boxes located further upstream. It is shown here that the housekeeping gene promoter-like region of human theta 1 contains two tandemly arranged, GC-rich motifs (GC-I and GC-II). Of these, GC-II interacts with nuclear factor(s) present in the globin-expressing, erythroleukemia cell line K562, before and after hemin induction. GC-I, however, interacts with nuclear factor(s) only present in hemin-induced K562 cells. These factors are different from previously reported erythroid cell-specific factors, and are not detectable in non-erythroid Hela cells. Furthermore, the sequence of the motif GC-I and its location relative to ATG codon have been conserved among all known mammalian theta 1 globin genes. Finally, and most interestingly, the CCAAT box of theta 1 is contained within a 38 bp internal segment of Alu repeat sequence. Immediately upstream from this CCAAT box-containing Alu repeat segment is a 241 bp Alu repeat pointing in the opposite direction. The conservation of this novel arrangement among the higher primates suggests that an inserted Alu family repeat and its flanking genomic sequence have co-evolved, for at least 30 million years, to provide the canonical CCAAT and TATA promoter elements of the theta 1 globin genes in higher primates.

Animals

The rabbit CD1 and the evolutionary conservation of the CD1 gene family.

A comparison of the genes encoding the CD1 leucocyte differentiation antigens in man and mouse shows important differences which prompted us to analyze the CD1 genes of the rabbit. We have found that the rabbit genome contains multiple CD1 loci. Upon cloning and sequencing, one of these loci was found to encode the known rabbit CD1-like antigen (R-Ta) and to be closely related to the human CD1b gene, which is absent in the mouse, while a second rabbit gene is closely related to both the human R3 and the mouse CD1 genes. The data reinforce the notion of the existence of two classes of CD1 genes, one of which is conserved in all species, while the other, albeit also evolutionarily old, has been deleted in mice as well as in other rodents.

Amino Acid Sequence

MR imaging of cavernous sinus involvement by pituitary adenomas.

The ability of high-resolution MR imaging (1.5 T) to detect invasion of the cavernous sinuses by pituitary adenoma was determined through a retrospective review of 74 patients. These patients were divided into three groups: 25 normal subjects, 24 subjects with invasive pituitary adenomas, and 25 subjects with noninvasive pituitary adenomas. A fourth group of 30 patients, who subsequently underwent surgery for pituitary adenoma, was evaluated prospectively by MR for the presence or absence of cavernous sinus invasion. Several features were analyzed: (1) the detectability of the medial and lateral dural margins of the cavernous sinus (2) the size and variation in intensity of compartments within the cavernous sinus (3) the relationship of endocrine function to the surgical and MR appearance of the cavernous sinus and (4) carotid artery displacement or encasement by tumor. The normal cavernous sinuses were usually symmetric, but their sizes varied. The lateral dural margin of the cavernous sinus was always recognized on MR as a linear, discrete, low-intensity area. The medial dural margin (pituitary capsule) was seen on MR in only two of the 25 normal patients. In all 24 patients with cavernous sinus invasion involvement was unilateral and was most common with laterally positioned prolactin or adrenocorticotropic hormone secretory adenomas. Invasion of the cavernous sinus was suspected by MR in only two of the 13 invasive microadenomas and was questionable in three. In 10 of the 11 macroadenomas with surgically proved dural invasion, MR demonstrated an asymmetric increase in size and intensity of the superior and inferior cavernous sinus compartments. Noninvasive macroadenomas compressed and displaced the cavernous sinus bilaterally. The prospective MR evaluation of 30 patients undergoing surgery for pituitary tumor revealed a sensitivity for predicting cavernous sinus invasion of 55%, a specificity of 85.7%, a positive predictive value of 62.5%, and a negative predictive value of 81.8%. No feature permitted certain distinction between invasive and noninvasive microadenomas, as the medial dural wall of the cavernous sinus could not be reliably identified. The most specific sign of cavernous sinus invasion was carotid artery encasement.

Adenoma

Reduction of common carotid resistance upon stimulation of an area dorsal to the facial nucleus of cats.

In chloralose-urethane-anesthetized cats, electrical stimulation and glutamate injection on a small reticular area just dorsal to the facial nucleus (DFA) elicited an ipsilateral reduction in the common carotid resistance (CCR-reduction) with no or minimal change in other cardiovascular parameters. CCR-reduction was mediated via facial and glossopharyngeal nerves, involving partially muscarinic and partially non-muscarinic mechanisms.

Animals

Molecular characterization of the HLA-linked steroid 21-hydroxylase B gene from an individual with congenital adrenal hyperplasia.

21-Hydroxylase deficiency which causes congenital adrenal hyperplasia is one of the most common defects of adrenal steroidogenesis. There are two 21-hydroxylase genes in man, A and B, and these have been mapped to the HLA class III region. Only the 21-hydroxylase B gene is thought to be active. To understand the molecular basis of congenital adrenal hyperplasia in a patient with the salt-wasting form of the disease, we cloned and characterized his single 21-hydroxylase B gene. The nucleotide sequence of this gene and a 21-hydroxylase B gene from a normal individual have been determined. Comparison of the two sequences has revealed 11 nucleotide alterations, of which two are in the 5' flanking region, four are in introns, one is in the 3' untranslated region and four are in exons. Two of the differences in exons cause codon changes, with Ser-269 and Asn-494 in the normal 21-hydroxylase B gene being converted to Thr and Ser, respectively. These amino acid substitutions may give an insight into those residues necessary for 21-hydroxylase enzymatic activity. We have also confirmed that the 21-hydroxylase A gene is a pseudogene due to three deleterious mutations in the exons. In addition, comparison of the 21-hydroxylase B gene sequence with other published sequences indicates that this microsomal cytochrome P-450 may be polymorphic.

Adrenal Hyperplasia, Congenital

Definitive RFLPs to distinguish between the human complement C4A/C4B isotypes and the major Rodgers/Chido determinants: application to the study of C4 null alleles.

Definitive restriction fragment length polymorphisms (RFLPs) representing the exact locations responsible for isotypicity between the human complement components C4A and C4B, and their generally associated major Rodgers (Rg1) and Chido (Ch1) antigenic determinants, have been designed. By means of C4d-specific genomic probe for Southern blot analysis, a C4A gene can be defined by the presence of the 276 bp and 191 bp N1a IV fragments, while a C4B gene can be defined by a single 467 bp N1aIV fragment. In addition, an Rg1-expressing C4 gene can be represented by a 565 bp EcoO 109 fragment, and a Ch1-expressing C4 gene by a 458 bp EcoO 109 fragment, under the same conditions. All these polymorphic restriction fragments can be unambiguously and conveniently detected. In combination with the Taq I polymorphic patterns specific for the C4 loci and for the neighboring 21-hydroxylase genes, the nature and structure of the tandem C4,21-hydroxylase gene complex can be elucidated. In this study, it is inferred that the null allele of the HLA haplotype B44 DR6 C4A3 C4BQO is not a C4B allele, but probably encodes another C4A 3 allotype at the second C4 locus.

Alleles