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Biomedical subjects

D A Miller

Publications and source records attributed to D A Miller.

At least 109 records · Page 6Linked to original sources

Inheritance of ribosomal gene activity and level of DNA methylation of individual gene clusters in a three generation family.

Ribosomal gene activity and levels of DNA methylation were investigated by cytochemical and immunological methods in the nucleolar organizer regions (NORs) of individually recognised acrocentric chromosomes. Mendelian inheritance of ribosomal gene activity in a three generation family was demonstrated, together with consistent behaviour of individual gene clusters in different carriers, even when environmental conditions were changed. For most chromosomes, an inverse relationship between gene activity and the level of DNA methylation was observed. Exceptions were the two chromosomes 15 and chromosomes 13cp and 22p, all being strongly chromomycin-A3-positive in their short arms. These chromosomes bound to anti-5-MeC antibodies with differential frequencies in the different carriers. The possibility of involvement of repetitive GC-rich DNA in this behaviour is discussed.

Azacitidine↗

A chimpanzee-derived chromosome-specific alpha satellite DNA sequence conserved between chimpanzee and human.

We describe a cloned 2.7 kb alpha satellite sequence, Pan-3, from the pygmy chimpanzee (Pan paniscus) that specifically hybridizes in situ to chromosome 19 in the pygmy chimpanzee and to the homeologous human chromosome, no. 17. Using high stringency conditions of hybridization on Southern blots, this sequence hybridized to DNA from both species of chimpanzee (P. paniscus and P. troglodytes) and from human but not to DNA from gorilla (Gorilla gorilla) or orangutan (Pongo pygmaeus). Partial sequence analysis showed that Pan-3 and a previously described human chromosome 17-specific clone have up to 91% sequence identity. To our knowledge this is the highest sequence similarity reported between alphoid subsets from human and any other primate.

Animals↗

Comparative mapping of a gorilla-derived alpha satellite DNA clone on great ape and human chromosomes.

We have isolated an alpha satellite DNA clone, pG3.9, from gorilla DNA. Fluorescence in situ hybridization on banded chromosomes under high stringency conditions revealed that pG3.9 identifies homologous sequences at the centromeric region of ten gorilla chromosomes, and, with few exceptions, also recognizes the homologous chromosomes in human. A pG3.9-like alphoid DNA is present on a larger number of orangutan chromosomes, but, in contrast, is present on only two chromosomes in the chimpanzee. These results show that the chromosomal subsets of related alpha satellite DNA sequences may undergo different patterns of evolution.

Animals↗

Isolation of a variant family of mouse minor satellite DNA that hybridizes preferentially to chromosome 4.

Two cosmids (HRS-1 and HRS-2) containing mouse minor satellite DNA sequences have been isolated from a mouse genomic library. In situ hybridization under moderate stringency conditions to metaphase chromosomes from RCS-5, a tumor cell line derived from the SJL strain, mapped both HRS-1 and HRS-2 to the centromeric region of chromosome 4. Sequence data indicate that these cloned minor satellite DNA sequences have a basic higher order repeat of 180 bp, composed of three diverged 60-bp monomers. Digestion of mouse genomic DNA with several restriction enzymes produces a ladder of minor satellite fragments based on a 120-bp repeat. The restriction enzyme NlaIII (CATG) digests all the minor satellite DNA into three prominent bands of 120, 240, and 360 bp and a weak band of 180 bp. Thus, the majority of minor satellite sequences in the genome are arranged in repeats based on a 120-bp dimer, while the family of minor satellite sequences described here represents a rare variant of these sequences. Our results raise the possibility that there may be other variant families of minor satellites analogous to those of alphoid DNA present in humans.

Animals↗

Urokinase treatment of neonatal aortoiliac thrombosis caused by umbilical artery catheterization: a case report.

Umbilical artery catheterization is known to be associated with aortoiliac thrombosis in approximately 1% of newborns in whom catheters are placed. We describe a case of catheter-associated aortoiliac thrombosis in a newborn who was successfully treated by urokinase infusion. The infant was monitored by imaging studies followed by ultrasonography. The use of real-time ultrasonography enabled us to image this infant's aortoiliac thrombosis in an accurate, noninvasive manner and monitor the effectiveness of thrombolytic therapy. The fibrinolytic state was achieved with a combination of intravenous and intraarterial infusions of urokinase. Frequent measurement of prothrombin time, activated partial thromboplastin time, fibrinogen, and thrombin time was used to guide fibrinolytic therapy. We reviewed the literature for similar cases of aortoiliac thrombosis in newborns; some were managed by medical means, others by operation. The success rate of the two approaches appeared to be approximately equal. Therefore we recommend the less invasive approach first--urokinase therapy--to be followed by surgical intervention if thrombolysis is unsuccessful.

Aorta, Thoracic↗

Establishing a family practice: a model to determine economic feasibility.

The availability of adequate health care is a major concern of urban and rural citizens. Community leaders and hospital administrators attempting to recruit family physicians and health care providers considering a specific community need a method by which they can evaluate a community's potential for supporting a first or additional primary care physician. To develop this method, a detailed survey of family practices geographically dispersed throughout Oklahoma was conducted in 1989. Data collected from family physicians and their administrative staff reflected the volume of ambulatory and hospital visits and the direct and indirect costs of the practice over the previous 12 months. Using the fixed and operating cost data, as well as number of patient visits and patient care revenue, we designed a model to project the economic feasibility of establishing a family practice in a specific community. This model can be used to project the number of visits a community can generate for a prospective family physician, as well as the direct and indirect costs, gross revenue, and net income of the practice.

Adolescent↗

The cognitive context of examinations in psychiatry using Bloom's taxonomy.

Psychiatric practice involves complex thinking patterns. In addition to commanding a huge number of facts, the student must learn to manipulate factual knowledge to solve diagnostic problems, develop treatment plans, and critically evaluate those plans. This study demonstrates an empirical method for evaluating the level of cognitive processes tested in multiple choice examinations. Use of Bloom's taxonomy in evaluating test items demonstrated the majority of test items on a psychiatry clerkship examination and a resident in-training examination fell into the most basic cognitive level, that of simple recall. The utility of Bloom's taxonomy is discussed along with implications for medical education.

Cognition↗

Characteristics of elderly persons seen in an urban psychiatric emergency room.

With the aging of the U.S. population, an increase in the use of the psychiatric emergency room by elderly persons is expected. In this study of 118 elderly persons seen in a general hospital psychiatric emergency room, half had a history of psychiatric illness. A third appeared to have chronic organic brain disease, and a third had affective disorders. Misprescription or misuse of medications was implicated in the problems of 27 patients. The problems of most patients were not emergencies; most had experienced symptoms of their presenting complaint for more than six months, finally overtaxing the coping capacities of their caregivers. The authors discuss factors that prevent elderly persons from seeking help on their own and emphasize the need for earlier interventions with elderly patients and for education of the public and professionals about issues, such as overmedication, that are important in the care of elderly persons.

Aged↗

Differential binding of transforming growth factor-beta 1, -beta 2, and -beta 3 by fibroblasts and epithelial cells measured by affinity cross-linking of cell surface receptors.

A murine fibroblast cell line (AKR-2B clone 84A) and an epithelial cell line (BALB/MK) were compared for their ability to bind different transforming growth factor-beta (TGF beta) species. The results of competitive binding assays indicated that the epithelial cells had a higher affinity for TGF beta than the fibroblasts. This difference may be the basis for the sensitivity of epithelial cells to much lower concentrations of TGF beta than fibroblasts. Affinity cross-linking studies showed that both cell types express the three cell surface TGF beta-binding molecules that have been previously described for a variety of cell types. The complexity of these cell surface binding proteins was further evaluated using all possible combinations of radiolabeled ligands in competition with each of the three unlabeled TGF beta species. Differences in the ability of specific TGF beta types to compete with radiolabeled TGF beta 2 for binding to the type I and II receptors were observed, with TGF beta 1 being more potent for epithelial cells, and TGF beta 2 being more potent for fibroblasts. In addition, a difference in the ability of different TGF beta species to compete the [125I]TGF beta 3 from epithelial cell surface receptors was apparent. TGF beta 2 was not able to compete with [125I]TGF beta 3 for binding to the type II receptor at any concentration tested, while TGF beta 1 and TGF beta 3 were about equally potent in competition for this receptor type. These differences in cell surface receptor binding of structurally and biologically similar molecules may reflect different functions for these molecules.

Animals↗

Survey to estimate prevalence of Leptospira interrogans infection in mature cattle in the United States.

A total of 5,142 kidney tissue samples and 5,111 serum samples from mature cattle in 49 states and Puerto Rico were collected at slaughter. Age of cattle ranged from 1 to 16 years (mean, 6.6 years). Leptospires were isolated from 88 (1.7%) kidney tissues, and 2,493 (49%) sera contained antibodies against 1 or more of 12 Leptospira interrogans serovars. Leptospires were observed by immunofluorescence in 41 (0.8%) kidney tissues. Using agglutinin-absorption tests, 73 (83%) isolates were identified as serovar hardjo, 11 (12.5%) as serovar pomona, and 4 (4.5%) as serovar grippotyphosa. By use of restriction endonuclease analysis studies of chromosomal DNA, all isolates differed from reference serovars but were identical to strains previously isolated from cattle or swine in the United States. Of the serovar hardjo isolates, 85% were identical to restriction endonuclease analysis type (genotype) hardjo-bovis A and 11 (15%) were identical to genotype hardjo-bovis B. Serovar pomona isolates were identical to genotypes kennewicki A (64%) or kennewicki B (36%), and serovar grippotyphosa isolates were identical to the RM 52 strain. Isolation rates were significantly (P less than 0.001) higher for beef cattle than for dairy cattle and were higher (P less than 0.001) for bulls than for cows. Combined culture and immunofluorescence results indicated that 2% of mature cattle were renal carriers of leptospires.

Agglutination Tests↗

Relationships between prevalence of Leptospira interrogans in cattle, and regional, climatic, and seasonal factors.

On the basis of serologic test results and isolation of leptospires from mature cattle, distribution and prevalence of Leptospira interrogans serovars and genotypes were compared by state and region of the United States. Relationships between isolation rate and month of sample collection, mean regional temperature, and mean regional precipitation were examined. Isolation rate and seroprevalence were significantly (P less than 0.001) higher for southeastern, south central, and Pacific coastal regions than for other regions of the United States. Isolates of genotypes hardjo-bovis A and kennewicki A and B, and of serovar grippotyphosa appeared to be randomly distributed. Genotype hardjo-bovis B isolates came from a southern area of the country that extends from Georgia to New Mexico. To the authors' knowledge, this is the first recorded isolation of serovar hardjo from Hawaii. Although significant relationship was not documented between isolation rate and month or season of the year, seroprevalence for summer, fall, and winter was significantly (P less than 0.001) higher than that for spring. Regional isolation rate was related more to mean temperature (r = 0.83; P less than 0.05) than to mean precipitation amount (r = 0.34; P greater than 0.50).

Animals↗

Regulation of the expression of the serine dehydratase gene in the kidney and liver of the rat.

Serine dehydratase was induced in the kidneys of normal rats by the administration of either glucagon or dexamethasone. The increase in enzyme activity was associated with an increase in both enzyme protein and its mRNA, which were determined respectively by Western blot and RNA blot analysis. No apparent differences were observed between kidney and liver in the molecular weights of serine dehydratase proteins and the sizes of their mRNAs. Although kidney serine dehydratase was dramatically induced by either glucagon or dexamethasone, the liver enzyme was induced by glucagon but not by dexamethasone alone in the intact rat. On the other hand, liver serine dehydratase was induced in starvation, diabetes mellitus, and a high-protein diet. The kidney enzyme could not be induced under any of these conditions.

Animals↗

Differential expression of genes encoding TGFs beta 1, beta 2, and beta 3 during murine palate formation.

Transforming growth factor beta 1 (TGF beta 1) has been shown to have multiple effects on primary cultures of palate-derived cell types. We report the analysis, by in situ hybridization, of RNA expression for three different TGF beta isoforms (TGF beta 1, beta 2, and beta 3) during murine embryonic palate development. Differential expression of the three TGF beta genes is seen in the palatal shelves in mesenchymal and epithelial cells known to be involved in the morphogenesis of this organ. Taken together, these results suggest that the TGF beta s act as endogenous factors involved in the formation of the mammalian palate.

Animals↗

Chromosome-specific subsets of human alphoid DNA identified by a chromosome 2-derived clone.

We have cloned an alphoid DNA fragment, pBS4D, from the DNA of a human-hamster hybrid cell line containing chromosome 2 as its only cytologically detectable human component. Under high stringency conditions, pBS4D hybridized in situ mostly to chromosome 2 and to a lesser extent to chromosomes 18 and 20. Restriction analysis using the DNA from selected somatic hybrid cell lines revealed that the genomic organization of this alphoid DNA differs on each of these three chromosomes.

Animals↗

Chromosomal localization of seven members of the murine TGF-beta superfamily suggests close linkage to several morphogenetic mutant loci.

Chromosomal locations have been assigned to seven members of the TGF-beta superfamily using an interspecific mouse backcross. Probes for the Tgfb-1, -2, and -3, Bmp-2a and -3, and Vgr-1 genes recognized only single loci, whereas the Bmp-2b probe recognized two independently segregating loci (designated Bmp-2b1 and Bmp-2b2). The results show that the seven members of the TGF-beta superfamily map to eight different chromosomes, indicating that the TGF-beta family has become widely dispersed during evolution. Five of the eight loci (Tgfb-1, Bmp-2a, Bmp-2b1, Bmp-2b2, Vgr-1) mapped near mutant loci associated with connective tissue and skeletal disorders, raising the possibility that at least some of these mutations result from defects in TGF-beta-related genes.

Amino Acid Sequence↗

HIV-1 reverse transcriptase purified from a recombinant strain of Escherichia coli.

A better understanding of the structure and biochemical properties of the replicative machinery of human immunodeficiency virus type 1 (HIV-1) may be useful in the screening and design of drugs that could be used to treat AIDS. We have previously described a recombinant strain of Escherichia coli that produces HIV-1 reverse transcriptase (RT). Fermentation conditions for the large-scale growth of the bacterial strain and a protocol for the purification of an enzymatically active 66-Kd form of the RT have been developed. The purified RT has all of the appropriate enzymatic functions and properties. The recombinant protein can be substituted for the viral enzyme in structural and biochemical studies and used in screens for drugs that could inhibit HIV replication.

Biotechnology↗

The effect of clerkship site on medical student examination grades.

Fairness and equity in grading medical students is a subject of continuing interest and concern. It appeared that the students on one clerkship site, the emergency room, were scoring less well on the written final examination than those on the other clerkship sites. Correlating the scores confirmed this impression. A simple intervention was made, and follow-up examination of the scores revealed that it was effective; in fact, the students on the emergency clerkship are now scoring higher than those on the other sites.

Achievement↗