Search PubMed⌕ Search

Biomedical subjects

D C Lynch

Publications and source records attributed to D C Lynch.

At least 37 records · Page 2Linked to original sources
PubMed1987

Molecular cloning of the human gene for von Willebrand factor and identification of the transcription initiation site.

A series of overlapping cosmid genomic clones have been isolated that contain the entire coding unit of the human gene for von Willebrand factor (vWf), a major component of the hemostatic system. The cloned segments span approximately 175 kilobases of human DNA sequence, and hybridization analysis suggests that the vWf coding unit is approximately 150 kilobases in length. Within one of these clones, the vWf transcription initiation site has been mapped and a portion of the vWf promoter region has been sequenced, revealing a typical "TATA box," a downstream "CCAAT box," and a perfect downstream repeat of the 8 base pairs containing the transcription start site. Sequencing of a segment of another genomic clone has revealed the vWf translation termination codon. Where tested, comparative restriction analysis of cloned and chromosomal DNA segments strongly suggests that no major alterations occurred during cloning and that there is only one complete copy of the vWf gene in the human haploid genome. Similar analyses of DNA from vWf-producing endothelial cells and nonexpressing leukocytes suggest that vWf gene expression is not accompanied by gross genomic rearrangements. In addition, there is significant homology of C-terminal coding sequences among the vWf genes of several vertebrate species.

Amino Acid Sequence↗
PubMed1987

Expression of abnormal von Willebrand factor by endothelial cells from a patient with type IIA von Willebrand disease.

Studies were conducted to characterize the biosynthesis of von Willebrand factor (vWf) by cultured endothelial cells (EC) derived from the umbilical vein of a patient with type IIA von Willebrand disease. The patient's EC, compared with those from normal individuals, produced vWf that had decreased amounts of large multimers and an increase in rapidly migrating satellite species, features characteristic of plasma vWf from patients with type IIA von Willebrand disease. The type IIA EC did produce a full spectrum of vWf multimers in both cell lysates and postculture medium, although the relative amounts of the largest species were decreased. The large multimers were degraded in conjunction with the appearance of rapidly migrating satellites that contained approximately equal to 170-kDa proteolytic fragments, suggesting that this patient's functional defect is due to abnormal proteolysis and not to a primary failure of vWf subunit oligomerization. Moreover, the observed degradation appears to result from an abnormal vWf molecule and not elevated protease levels. These results suggest that this patient's von Willebrand disease phenotype is caused by increased proteolytic sensitivity of his vWf protein.

Cell Division↗
PubMed1986

An explanation for minor multimer species in endothelial cell-synthesized von Willebrand factor.

Initial synthesis of von Willebrand factor (vWf) by cultured human endothelial cells proceeds by formation of a dimer of pro-vWf subunits. These subunits are found only within the cell and have an apparent molecular weight of 240,000-260,000, as measured by electrophoresis in sodium dodecyl sulfate-polyacrylamide gels. Posttranslational modifications, including proteolytic cleavage, glycosylation, and sulfation, result in the appearance of two additional vWf subunits. The major one migrates with the subunit of plasma vWf at an apparent molecular weight of 220,000-225,000 and the other migrates more slowly than pro-vWf at an apparent molecular weight of 260,000-275,000. These subunits oligomerize to form a set of vWf multimers, which are subsequently secreted into the culture medium. We isolated individual vWf oligomer species from the agarose gel bands and show that vWf minor, or satellite, species differ from major species in subunit composition.

Carbohydrate Metabolism↗
PubMed1985

Molecular cloning of cDNA for human von Willebrand factor: authentication by a new method.

We have identified a 2.4 kb partial cDNA clone (pDL34) for human von Willebrand factor (vWf) mRNA. pDL34 was selected by screening an endothelial cell cDNA library with a radiolabeled reverse transcript of mRNA obtained by specific immunoisolation of vWf polysomes from endothelial cells. pDL34 selectively hybridized to an endothelial cell-associated 9.5 kb mRNA. To confirm its identity, SP6 RNA polymerase was used to generate in vitro transcripts of the cDNA. This synthetic RNA, truncated at its 5' end, directed the synthesis of several unique polypeptides in rabbit reticulocyte lysates. These polypeptides were immunoprecipitated by polyclonal and monoclonal anti-vWf antibodies. These results indicate that pDL34 contains an authentic partial copy of vWf mRNA. In vitro transcription of partial cDNA clones and translation of the resulting RNAs may be a useful general method of verifying the identity of various cDNA clones in circumstances where antibodies are available and protein sequence is not.

Antibodies, Monoclonal↗
PubMed1983

Subunit composition of oligomeric human von Willebrand factor.

The oligomerization of human endothelial cell-synthesized von Willebrand factor (vWf) has been studied by gel chromatography in columns of Sephacryl S-500 and by discontinuous agarose gel electrophoresis. A quantitative recovery of high Mr vWf oligomers has been obtained after binding to a monoclonal anti-vWf-Sepharose adduct. This reagent has been used to analyze gel filtration chromatographic elution profiles of [35S]methionine-labeled culture medium and cell lysate. It was determined that high Mr oligomers are present in endothelial cell lysates as well as in the medium overlying these cells and are composed of Mr 225,000 subunits. When vWf oligomers were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the presence of a reducing agent, the Mr 240,000 subunit (provWf) was not observed to oligomerize beyond the dimer stage to a significant degree. Therefore, vWf oligomerization appears to be facilitated by conversion of provWf subunits to mature vWf subunits, most likely by proteolytic removal of sequences unique to the intracellular precursor.

Blood Coagulation Factors↗
PubMed1983

Fetal echocardiography. A tool for evaluation of in utero cardiac arrhythmias and monitoring of in utero therapy: analysis of 71 patients.

Fetal echocardiographic studies were performed in 71 patients referred for evaluation of cardiac rhythm disturbances at 24 to 40 weeks' gestation. After 2-dimensional echocardiographic study of cardiac structure was performed, M-mode echocardiograms were analyzed for measurement of cardiac rate, atrioventricular contraction sequence, atrioventricular valve motion, and duration of postectopic pauses. Arrhythmias were diagnosed in 59 patients. In 34 patients with isolated ectopic beats, the arrhythmia resolved during later pregnancy in 26 or within the first 5 days of life in 8. Six patients had mild sinus bradycardia and 8 had frequent sinus pauses; all 14 had resolution of the arrhythmia during pregnancy. Sustained arrhythmias occurred in 11 patients. Deaths occurred when there was associated fetal congestive heart failure (hydrops fetalis), structural heart disease, or both. M-mode echocardiography diagnosed supraventricular tachycardia in 3 fetuses. The echocardiogram was used thereafter for monitoring transplacental digoxin therapy.

Anti-Arrhythmia Agents↗
PubMed1983

Biosynthesis of the subunits of factor VIIIR by bovine aortic endothelial cells.

The biosynthesis of the subunit of factor VIIIR was studied in bovine aortic endothelial cells by the techniques of immunoprecipitation and NaDodSO4/polyacrylamide gel electrophoresis. It was determined that the subunit is first produced as a Mr 240,000 glycoprotein precursor, which appears to undergo proteolytic cleavage at or about the time of secretion into the medium with a resultant change in apparent Mr to 225,000, the size of the mature subunit found in plasma. The Mr 240,000 species was detected within 10 min of the start of labeling of cells, but factor VIIIR was not detected in the culture medium until approximately equal to 50 min. Treatment of the cells with tunicamycin resulted in a decrease in the apparent Mr of both species but did not grossly inhibit processing of precursor to product or the secretion of the latter. Thus, much or all N-linked glycosylation of factor VIIIR is not essential for these steps. Accumulation of factor VIIIR in the medium continued over a 24-hr period of cell labeling with [35S]-methionine, without significant net intracellular accumulation of the precursor, suggesting that a large storage pool of factor VIIIR is not present in endothelial cells under these conditions.

Animals↗
PubMed1982

Fetal echocardiography for evaluation of in utero congestive heart failure.

Thirteen fetuses with nonimmune hydrops (22 to 39 weeks of gestation) were evaluated with two-dimensional and M-mode echocardiography. Ten fetuses had cardiovascular abnormalities resulting in heart failure, and three had noncardiac causes of hydrops. In three cases, hydrops was caused by supraventricular tachycardia. One of these fetuses responded to cardioversion at birth, another responded to transplacental digoxin therapy, and the third died with atrial flutter and high-grade atrioventricular block before delivery. There were no cases of "idiopathic" hydrops. Our results show that fetal echocardiography is useful in determining cardiac causes of in utero heart failure resulting in hydrops fetalis. The fetal echocardiogram may also be used in monitoring transplacental therapy of heart failure.

Adolescent↗
PubMed1980

Prenatal echocardiography.

Congenital cardiac defects are not common, but their serious consequences make early detection and treatment essential. In developing a set of workable criteria for selective screening of serious abnormalities, the authors used commercially available ultrasound equipment in more than 500 high-risk pregnancies. Possible future applications include elucidation of causes of fetal cardiac defects.

Echocardiography↗
PubMed1980

Echocardiographic studies of the human fetus: prenatal diagnosis of congenital heart disease and cardiac dysrhythmias.

During obstetrical ultrasound examinations, 200 M-mode and 35 real-time two-dimensional echocardiographic studies were performed on 180 fetuses of high-risk pregnancies. Fetal gestational ages ranged from 18 to 41 weeks. M-mode "sweeps" demonstrating mitral- and septal-aortic fibrous continuity were obtained in 115 studies. Paradoxic septal motion in 50 fetuses suggested relarive right ventricular volume loading. Congenital cardiac malformations were accurately diagnosed in a 34-week fetus with pulmonary atresia and hypoplastic right ventricle and in a 28-week fetus with a univentricular heart. Congenital complete atrioventricular block was diagnosed in a 28-week fetus and atrial flutter with variable atrioventricular block was diagnosed in a 38-week fetus. The use of echocardiographic studies to evaluate cardiac structure and rhythm in utero assists in counseling prospective parents and in planning postnatal management for their offspring.

Arrhythmias, Cardiac↗
PubMedDate not supplied

Variables that may enhance medical students' perceived preparedness for computer-based testing.

OBJECTIVE: To identify variables that may enhance medical student's preparedness for computer-based administration of the United States Medical Licensing Examination (USMLE). DESIGN: A cross-sectional survey of 301 medical students who completed a self-administered questionnaire. MEASUREMENTS: The questionnaire was designed to obtain information about students' computer resources, personal experience with computers, computer expertise, opinions about computers, experience with computer-based testing, perceived preparedness for the computer-based USMLE, and demographic variables. Variables related to students' perceived preparedness for the computer-based USMLE were identified by ordinal logistic regression. RESULTS: A significant regression model yielded four significant predictors: perceived preparedness for USMLE content (P: < 0.0001), opinions about computers (P: < 0.0012), gender (P: < 0.0001), and a gender by computer-based testing experience interaction (P: < 0. 0004). Computer resources, personal experience with computers, computer expertise, age, race, and year of medical school were not significant predictors. CONCLUSION: Students' perceived preparedness for computer-based administration of high-stakes examinations may be facilitated by preparing them for examination content, by enhancing their opinions about computers, and by increasing their computer-based testing experiences.

Attitude to Computers↗
PubMedDate not supplied

Can a 3-day preceptorship change first-year medical students' opinions about living and working in small towns?

BACKGROUND: Research of medical school initiatives that attempt to orient medical students toward rural medicine may facilitate development of initiatives to alleviate physician maldistribution. This study investigated the effect of a 3-day family medicine preceptorship in a small town on first-year medical students' opinions about a) living and working in small towns and b) plans to live in and practice medicine in small towns. Student feedback about the preceptorship was also examined. METHODS: Questionnaires were administered to 137 first-year medical students using a separate sample pretest-posttest design. RESULTS: Student feedback indicated that the preceptorship was a valuable learning experience, but the preceptorship did not appear to influence students' opinions about or interest in living in and working in small towns or rural areas. CONCLUSION: Brief exposure to rural medicine early in the curriculum appears to have little effect on variables that might precede practice location decisions.

Adult↗