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F Gannon

Publications and source records attributed to F Gannon.

At least 55 records · Page 3Linked to original sources
PubMed1998

Identification of novel chicken estrogen receptor-alpha messenger ribonucleic acid isoforms generated by alternative splicing and promoter usage.

Using the rapid amplification of complementary DNA ends (RACE) methodology we have identified three new chicken estrogen receptor-alpha (cER alpha) messenger RNA (mRNA) variants in addition to the previously described form (isoform A). Whereas one of the new variants (isoform B) presents a 5'-extremity contiguous to the 5'-end of isoform A, the two other forms (isoforms C and D) are generated by alternative splicing of upstream exons (C and D) to a common site situated 70 nucleotides upstream of the translation start site in the previously assigned exon 1 (A). The 3'-end of exon 1C has been located at position -1334 upstream of the transcription start site of the A isoform (+1). Whereas the genomic location of exon 1D is unknown, 700 bp 5' to this exon were isolated by genomic walking, and their sequence was determined. The transcription start sites of the cER alpha mRNA isoforms were defined. In transfection experiments, the regions immediately upstream of the A-D cER alpha mRNA isoforms were shown to possess cell-specific promoter activities. Three of these promoters were down-regulated in the presence of estradiol and ER alpha protein. It is concluded, therefore, that the expression of the four different cER alpha mRNA isoforms is under the control of four different promoters. Finally, RT-PCR, S1 nuclease mapping, and primer extension analysis of these different cER alpha mRNA isoforms revealed a differential pattern of expression of the cER alpha gene in chicken tissues. Together, the results suggest that alternative 5'-splicing and promoter usage may be mechanisms used to modulate the levels of expression of the chicken ER alpha gene in a tissue-specific and/or developmental stage-specific manner.

Alternative Splicing↗
PubMed1998

Differentially expressed messenger RNA isoforms of the human estrogen receptor-alpha gene are generated by alternative splicing and promoter usage.

The isolation and characterization of several new human estrogen receptor-alpha (hERalpha) mRNAs are described. Together with those previously identified, they give rise to a total of six hERalpha mRNA isoforms (A-F hERalpha mRNAs). Produced from a single hERalpha gene by multiple promoter usage, all these transcripts encode a common protein but differ in their 5'-untranslated region as a consequence of alternative splicing of five upstream exons (1B-1F). RT-PCR and S1 nuclease mapping analysis of these different hERalpha mRNA isoforms revealed a differential pattern of expression of the hERalpha gene in human tissues and cell types. The A hERalpha mRNA is the main isoform detected in mammary glands or in the tumor cell lines derived from this tissue. In endometrium, the predominant forms are the A and C hERalpha mRNA isoforms, whereas the C and F hERalpha mRNA isoforms are the major forms detected in ovary. Finally, high levels of the E hERalpha mRNA isoform are restricted to the liver with an increased expression in females. Taken together, our results demonstrate that the hERalpha gene is a complex genomic unit exhibiting alternative splicing and promoter usage in a tissue-specific manner.

Adolescent↗
PubMed1998

Urinary basic fibroblast growth factor. A biochemical marker for preosseous fibroproliferative lesions in patients with fibrodysplasia ossificans progressiva.

Angiogenesis is a prominent histopathologic feature of preosseous fibroproliferative lesions in patients who have fibrodysplasia ossificans progressiva. Basic fibroblast growth factor is an extremely potent in vivo stimulator of angiogenesis, and has been implicated in the growth of solid tumors. An enzyme linked immunosorbent assay for basic fibroblast growth factor was performed on urine samples from patients who had active (n = 28) and inactive (n = 39) fibrodysplasia ossificans progressiva, and compared with urine samples from normal age and gender matched control subjects (n = 54). Median basic fibroblast growth factor levels were 2705 pg/g of creatinine in the normal control group, 5058 pg/g of creatinine in patients with inactive fibrodysplasia ossificans progressiva (no significant difference), and 8793 pg/g of creatinine in patients with active fibrodysplasia ossificans progressiva. Female subjects, both normal and with fibrodysplasia ossificans progressiva, had higher levels of urinary basic fibroblast growth factor than did male subjects. There was no correlation of urinary basic fibroblast growth factor levels with age or severity of preexisting disability. These data document an elevation of urinary basic fibroblast growth factor during acute flareups of fibrodysplasia ossificans progressiva and provide a biochemical basis for considering antiangiogenic therapy for inhibiting endochondral osteogenesis in this disorder.

Adolescent↗
PubMed1997

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Academies and Institutes↗
PubMed1997

Sequence analysis and tissue expression of a non-Bohr beta-globin cDNA from Atlantic salmon.

The presence of a haemoglobin protein which does not exhibit a Bohr effect has been found only in fish living in fast flowing waters. We report the cloning of the first non-Bohr effect beta-globin cDNA from an adult Atlantic salmon kidney bank. Nucleotide sequence analysis of this cDNA shows that the predicted beta-globin peptide comprises 147 amino acids with a calculated molecular mass of 15 975 Da and an overall amino acid homology of 40 to 50% to higher vertebrates and 60-90% to fish sequences. This sequence confirms the important amino acid residues which are changed thus causing loss of the Bohr effect [Powers, D.A. and Edmunson, A.B. (1972) Multiple hemoglobins of catostomid fish. J. Biol. Chem. 247, 6686-6693; Brunori, M. (1975) Molecular adaptation to physiological requirements: the hemoglobin system of trout. Curr. Topics Cell. Regul. 9, 1-39]. This loss allows the haemoglobin protein to have a higher oxygen affinity, as it does not release oxygen when the pH of the surrounding environment decreases, which is an important ability for the fish in times of stress.

Amino Acid Sequence↗
PubMed1997

Improved efficiency for primer extension by using a long, highly-labeled primer generated from immobilized single-stranded DNA templates.

Primer extension is one of the most common methods used to measure the amount and size of RNAs. We demonstrate that the sensitivity and the specificity of this method are improved considerably by using a highly-labeled single-stranded DNA generated from a biotinylated single-stranded DNA template, as a long specific primer in the reverse transcription reaction. This new approach allows the detection of transcripts with a low expression level from microgram quantities of total RNA.

Animals↗
PubMed1997

Regional differences of dura osteoinduction: squamous dura induces osteogenesis, sutural dura induces chondrogenesis and osteogenesis.

Dura plays an important role in calvarial morphogenesis. However, precisely what that role is remains unclear. We present here in vivo evidence that dura without other central nervous system components induces both chondrogenesis and osteogenesis. The mechanism is, at least in part, by proximate tissue interaction. The objectives of this experiment were to answer the following: (1) Can dura actually induce osteogenesis without the influence of the underlying brain? (2) What are the requirements of this dura-induced heterotopic osteogenesis? (3) What are the differences between dura underlying sutures and dura underlying the squamous portions of the cranial bones? Dura underlying the metopic, sagittal, and lambdoidal sutures and dura underlying the flat portions of frontal and parietal bones were obtained from neonatal Lewis rats and transplanted into the posterior thoraces of adult Lewis recipients. In group I, dura underlying the metopic, sagittal, and lambdoidal sutures (n = 20) and dura underlying the flat portions of frontal and parietal bones (n = 20) were transplanted individually into separate epitheliomesenchymal pockets. Group II animals had dura underlying the metopic, sagittal, and lambdoidal sutures (n = 10) and dura underlying the flat portions of frontal and parietal bones (n = 10) transplanted individually into surgically created mesenchymal pockets by placing the dura grafts between panniculus carnosus and latissimus dorsi muscles. The animals were sacrificed at 2-week intervals. Light microscopy, special histochemical analysis, immunohistochemistry, and electron microscopy were performed. Bone formation was seen in 15 of the 18 animals (83 percent) in group I. No bone or cartilage formation was seen in group II. Chondrogenesis was seen in 4 animals receiving dura underlying the metopic, sagittal, and lambdoidal sutures in group I. Cellular hyperproliferation was seen at 2 weeks when dura was transplanted close to the hair follicles. These cells had a high nucleus-to-cytoplasm ratio and were positive for transforming growth factor beta. This hyperproliferation was followed by production and accumulation of Alcian blue-positive extracellular matrix that resisted digestion by hyaluronidase. Cellularly active cartilage was seen at 6 weeks. There was no chondrogenesis in animals receiving dura underlying the flat portions of frontal and parietal bones in group I. Electron microscopy demonstrated the presence of proteoglycan-like ground substance and type II collagen in the inner layer of sutural dura and the predominance of dense type I collagen in the squamous dura and the external layer of the sutural dura. The important findings of this experiment are that (1) heterotopically transplanted neonatal dura can induce osteogenesis, (2) this heterotopic osteoinduction by dura requires epitheliomesenchymal interaction, and (3) separating dura into sutural dura and squamous dura, chondrogenesis occasionally occurred in addition to osteogenesis with the former, while only membranous ossification occurred with the latter, indicating intrinsic differences within the dura mater. This dural heterogeneity is supported by direct ultrastructural data.

Animals↗
PubMed1996

Hepatocyte nuclear factor 4 (HNF4) binding sites in the salmon HNF1 promoter.

We report the isolation and sequencing of a 1100-bp DNA fragment containing the salmon Hepatocyte Nuclear Factor 1 gene (sHNF1) promoter. The sHNF1 promoter cloned upstream of the chloramphenicol acetyl transferase (CAT) encoding gene is shown to be active in two cell lines of hepatic origin. DNasel footprint analysis of the proximal 400 bp reveals several protein-binding sites, including a CCAAT box, a potential site for Sp1, and three potential HNF4 binding sites. The sequence does not contain any canonical TATA box or initiator and sHNF1 transcription is initiated at four different sites spanning a region of 56 bp. Sequence comparison with the Xenopus laevis HNF1 promoter sequence did not show any significant similarity except in the region overlapping two of the potential HNF4 binding sites.

Animals↗
PubMed1996

A Mycobacterium malmoense-specific DNA probe from the 16S/23S rRNA intergenic spacer region.

Mycobacterium malmoense was first described in 1977. It is now recognized as an opportunistic human pathogen which can be difficult to identify using standard methods. M. malmoense may be underestimated as the causative agent of clinical disease because of the recognized difficulties in its primary cultivation and identification. In this study, the nucleotide sequence of the 16S/23S rRNA intergenic spacer region from five clinical isolates of M. malmoense has been determined, in order to develop a PCR-based DNA probe assay to facilitate the early identification of this organism. The DNA sequence generated was utilized to design an oligunucleotide probe that specifically hybridizes with M. malmoense. The ability of this DNA probe to detect geographically distinct M. malmoense isolates was investigated. The value of this DNA probe was realized by its ability to differentiate three isolates of the Mycobacterium avium complex, which had been misidentified as M. malmoense using conventional biochemical methods.

Base Sequence↗
PubMed1996

Chondromalacia patellae: an in vitro study. Comparison of MR criteria with histologic and macroscopic findings.

OBJECTIVE: To develop MR criteria for grades of chondromalacia patellae and to assess the accuracy of these grades. DESIGN: Fat-suppressed T2-weighted double-echo, fat-suppressed T2-weighted fast spin echo, fat-suppressed T1-weighted, and gradient echo sequences were performed at 1.5 T for the evaluation of chondromalacia. A total of 1000 MR, 200 histologic, and 200 surface locations were graded for chondromalacia and statistically compared. RESULTS: Compared with gross inspection as well as with histology the most accurate sequences were fat-suppressed T2-weighted conventional spin echo and fat suppressed T2-weighted fast spin echo, although the T1-weighted and proton density images also correlated well. The most accurate MR criteria applied to the severe grades of chondromalacia, with less accurate results for lesser grades. CONCLUSIONS: This study demonstrates that fat-suppressed routine T2-weighted and fast spin echo T2-weighted sequences seem to be more accurate than proton density, T1-weighted, and gradient echo sequences in grading chondromalacia. Good histologic and macroscopic correlation was seen in more severe grades of chondromalacia, but problems remain for the early grades in all sequences studied.

Cadaver↗
PubMed1996

Structural organization and sequence analysis of the globin locus in Atlantic salmon.

Preliminary analysis of Atlantic salmon alpha- and beta-globin genes indicated that these genes are linked in a 3' to 3' orientation, with the RNA-coding sequences located on opposite strands. In this report, we show that two different alpha-globin genes have the same orientation and are encoded on the same strand whereas two different beta-globin genes are encoded on the opposite strand and also have the same orientation. This cluster of globin genes is divided into two subclusters: one for the Bohr globin genes and one for the non-Bohr globin genes. This is the first evidence for this type of arrangement found for globin genes. DNase I footprint analysis of two of the globin promoters show erythroid-specific transcription factor binding sites that have also been found in human and other mammalian globin genes.

Animals↗
PubMed1996

An experimental study of the effects of craniofacial growth on the long-term positional stability of microfixation.

Recent clinical reports have documented a potential problem associated with the use of microfixation in the growing craniofacial skeleton, i.e., the finding of microplates and microscrews within the endocranial cavity during repeat surgery. This animal experiment using Yorkshire pigs was designed to determine if this type of passive internal displacement could be demonstrated in an animal model, the frequency of internal displacement, and the potential consequence of such events from a neurohistological standpoint. Ten female Yorkshire piglets underwent left unilateral fronto-orbital advancement at age 3 weeks, fixation being achieved with microplates and screws. The position of the hardware was determined by direct inspection, after they were killed at 6 months of age (skeletal maturity). Ten microplates (28%) showed complete intracranial translocation, whereas nine (27%) remained on the ectocranial surface. The majority, 16 (44%), were between the outer and inner cortices of the calvaria. Gross examination of the brain tissue underlying the translocated hardware showed indentation "pits" forming as a result of such translocation. Histological examination of the cerebrum and meninges of these pits revealed loss of the most superficial connecting cell layer, with focal neuronal distortion. The leptomeninges were intact and there was no cerebritis, gliosis, or hypoxic change. These findings confirm that in this pig model microfixation hardware will undergo passive intracranial ranial translocation and that there are demonstrable histological alterations in the underlying brain and meninges. The long-term effects of these observed anatomical changes are unknown.

Animals↗
PubMed1995

Severe osteoporosis in men.

OBJECTIVE: To evaluate men with severe osteoporosis for pathogenetic factors and to review the reported features of primary osteoporosis in men. DESIGN: Case series and clinical review. PATIENTS: 47 men consecutively referred to a metabolic bone center because of atraumatic (or minimally traumatic) fractures (91%) or radiographic osteopenia (9%). MEASUREMENTS: Clinical assessment, radiographs, chemical analyses of serum and urine, hormone assays, skeletal densitometry, and histomorphometry of iliac crest biopsy specimens. RESULTS: 27 of the 47 men (57%) had vertebral fractures, and 16 (34%) had appendicular fractures. Causal factors identified in 30 men (64%) included glucocorticosteroid treatment (8 men); hypogonadism (7 men); excessive alcohol consumption (7 men); and anticonvulsant use, osteomalacia, severe hyperthyroidism, or bone marrow neoplasia (8 men). Seventeen men (36%) had no medical conditions or known risk factors associated with bone disease. Spinal mineral density was well below the mean value for healthy young men in 94% of the patients with primary osteoporosis tested. Examination of biopsy specimens from 13 of 17 men with primary osteoporosis showed reduced trabecular bone volumes, normal bone formation rates, and slightly increased resorption surfaces. Fasting hypercalciuria was seen in some men (41%). In the primary osteoporosis group, eight men were followed serially (range of follow-up, 6 months to 9 years) while they were receiving a nonpharmacologic regimen (diet and activity); the mean axial bone mineral density of these men increased slightly. CONCLUSIONS: A thorough evaluation for identifiable causes of severe osteoporosis in men is warranted because definable pathogenetic factors are seen in many cases. A few men with severe osteoporosis have primary or idiopathic osteoporosis. Primary osteoporosis in men is probably caused by many factors because heterogeneous clinical, laboratory, and histologic features were seen in our series and in those of others. Further studies of primary osteoporosis are needed to define the course of the disease, to identify pathogenetic mechanisms, and to develop therapeutic interventions.

Adult↗
PubMed1995

Presence of an extended duplication in the putative low-density-lipoprotein receptor-binding domain of apolipoprotein B. Cloning and characterization of the domain in salmon.

The sequence of the C-terminal 1058 amino acids of atlantic salmon (Salmo salar) apolipoprotein (apo) B was deduced from the nucleotide sequence of cloned cDNA. In comparison with chicken or mammals apoB-100, salmon apoB is C-terminally truncated and extended gaps are found. The two clusters of positively charged residues, previously identified as part of the putative low-density-lipoprotein (LDL) receptor-binding domain of apoB, are brought into close proximity in salmon apoB. This is achieved by the absence between the two clusters of the proline-rich area with the potential to form an amphipathic beta sheet, present in higher vertebrates. In addition, analysis of apoB amino acid sequences currently available in vertebrates revealed the presence of an extended internal duplication in the putative LDL receptor-binding domain. Thus, the two basic clusters would have been duplicated resulting in the presence, except for salmon apoB, of two homologous sites in the C-terminal part of the molecule. The results described here together with earlier biochemical and genetic evidence support the view that Arg3500, a residue mutated in familial defective apoB-100, could be included in a folded critical region of the putative LDL receptor-binding domain of human apoB-100. This region possibly brings the two sub-domains that arise from the duplication close to each other.

Amino Acid Sequence↗