Search PubMed⌕ Search

Biomedical subjects

D M Miller

Publications and source records attributed to D M Miller.

At least 91 records · Page 5Linked to original sources
PubMed1995

Expression of beta-galactosidase under the control of the human c-myc promoter in transgenic mice is inhibited by mithramycin.

In order to assess the functional contribution of the human c-myc promoter region in the expression of the c-myc gene, transgenic mouse lines containing a bacterial lac Z gene encoding beta-galactosidase under the control of the human c-myc protooncogene promoter were generated. Transgenic mouse embryos heterozygous for the human c-myc Z transgene demonstrate high amounts of beta-galactosidase activity as early as day 11 of embryogenesis by histochemical staining of whole embryos using 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-Gal) as substrate, localizing specifically to early spinal cord tissue. beta-galactosidase activity can be demonstrated by histochemical staining in brain tissue of day 14 embryos, localizing mainly to the prefrontal cortex region, while relative amounts of beta-galactosidase in spinal cord tissue are reduced. Determination of specific activity of beta-galactosidase using resorufin-beta-galactopyranoside as substrate in homogenates of whole embryos heterozygous for the human c-myc/lac Z transgene demonstrates significantly elevated beta-galactosidase activity over control embryos in day 11 and day 14 embryos. Surprisingly, cell homogenates of brain tissue from adult G1 generation mice heterozygous for the human c-myc/lac Z transgene demonstrate greater than 10-fold higher specific activity of beta-galactosidase over normal control brain tissue. Specific inhibition of the c-myc/lac Z transgene was also demonstrated in developing embryos using mithramycin given at a dose of 150 micrograms kg-1 d-1 intraperitoneal to pregnant females on days 7-13 of gestation. Both histochemical staining of beta-galactosidase and specific activity assays of day 14 embryos demonstrated significantly lower levels of beta-galactosidase than untreated controls. These results are unique since we are able to detect expression of beta-galactosidase in developing embryonic central nervous system tissue along with adult brain tissue of animals carrying the human c-myc Z transgene and we are able to specifically inhibit expression of the transgene using mithramycin administered in utero.

Animals↗
PubMed1995

Inhibition of nuclear protein binding to two sites in the murine c-myc promoter by intermolecular triplex formation.

The c-myc gene is overexpressed in a variety of tumor types and appears to play an important role in the abnormal growth of a number of cell types. In an effort to determine the ability of sequence- and species-specific triplex-forming oligonucleotides to inhibit expression of a targeted gene in animals, we have identified two novel triplex-forming sites in the murine c-myc promoter. One is homologous to the triplex-forming human PuF binding element located upstream of the P1 transcription start site. The other triplex-forming site is found in a region between P1 and P2 that encompasses the ME1a1 binding site and part of the E2F binding site and is highly homologous to the human sequence. Synthetic oligodeoxyribonucleotides designed to target these essential regulatory elements form sequence-specific triple helices as demonstrated by gel mobility shift analysis and DNase I footprinting. Polypurine: polypyrimidine regions in the P1 and P2 promoters form specific protein-DNA complexes upon incubation with a murine YC8 nuclear extract. Preincubation of each of the promoter fragments with its respective triplex-forming oligonucleotide results in the inhibition of nuclear protein binding. Non-triplex-forming oligonucleotides do not significantly affect protein binding. The data presented are a preliminary step toward generating an animal model for the phenotypic effects of triplex formation within the c-myc promoter.

Animals↗
PubMed1995

The c-myc promoter binding protein (MBP-1) and TBP bind simultaneously in the minor groove of the c-myc P2 promoter.

The c-myc promoter binding protein (MBP-1) is a DNA binding protein which negatively regulates the expression of the human c-myc gene. MBP-1 binds to a sequence which overlaps the binding site for the general transcription factor TBP, within the c-myc P2 promoter region. Since TBP binds in the minor groove, MBP-1 might inhibit c-myc transcription by preventing the formation of a functional preinitiation complex. In support of this hypothesis, we have demonstrated that MPB-1 is a minor groove binding protein. In order to characterize MBP-1 binding, we substituted A-T base pairs in the MBP-1 binding site with I-C base pairs, which changes the major groove surface without altering the minor groove surface. This substitution did not inhibit the sequence-specific binding of MBP-1 and TBP. On the other hand, G-C to I-C substitution within the MBP-1 binding site alters the minor groove and prevents MBP-1 binding. Competitive electrophoretic mobility shift assays were used to show that berenil, distamycin, and mithramycin, all of which bind in the minor groove, compete with MBP-1 for binding to the MPB-1 binding site. These minor groove binding ligands also effectively inhibit the simultaneous DNA binding activity of both MBP-1 and TBP. We conclude that both MBP-1 and TBP can bind simultaneously in the minor groove of the TATA motif on the c-myc P2 promoter. This suggests that MBP-1 may negatively regulate c-myc gene expression by preventing efficient transcription initiation.

Base Sequence↗
PubMed1995

Epidermal growth factor secreted from the salivary gland is necessary for liver regeneration.

Partial hepatectomy (PH) in rats induces a synchronized burst of DNA replication in the remnant liver that peaks at 24 h post-PH. We report here that removal of the major salivary glands before one-third and two-thirds PH prevents the proliferative response in the remaining liver. Twelve days after one-third PH, the remnant liver is 89% of the normal liver weight in nonsalivectomized rats but only 55% in salivectomized animals. This indicates that salivectomy does not merely delay the first round of cell division but that it prevents actual regeneration. Salivectomy alters the early protooncogene response to partial hepatectomy. In salivectomized rats, the characteristic peak of c-myc mRNA synthesis at 2-4 h after PH is significantly decreased compared with nonsalivectomized rats. The peak of DNA synthesis at 24 h after PH in salivectomized rats is also dramatically decreased. DNA synthesis as measured by [3H]thymidine incorporation into DNA of hepatic cells is decreased approximately 90% in salivectomized rats vs. nonsalivectomized rats 22-26 h after PH. Ligation of the venous drainage of the salivary gland results in the same inhibitory effect on DNA synthesis, indicating 1) that the salivary gland must release circulating factor(s), and 2) that the early increase in c-myc expression and the subsequent DNA synthesis, both of which reflect the stimulation of cellular proliferation in the regenerating liver, are induced by humoral factor(s) released from the salivary glands. Injection of exogenous epidermal growth factor (EGF) in salivectomized rats results in restoration of both the DNA synthetic and c-myc responses at levels characteristic of those of liver regeneration.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Breathing systems reclassified.

The purpose of reclassifying breathing systems is to enhance understanding by relating structure to function. A previous classification which appears to fulfill this objective is updated to include recent developments in a newly named group of systems called "displacement afferent reservoir" (DAR) breathing systems. A classification table allows for quick comparison and shows that, of the non-absorber systems, the DAR group has the best characteristics of fresh gas utilization.

Absorption↗
PubMed1995

An enclosed efferent afferent reservoir system: the Maxima.

A single, lightweight, valveless, non-absorber breathing system, which functions with near maximal efficiency (hence the name Maxima) in spontaneous and controlled ventilation is described. It may be classified as an enclosed efferent afferent reservoir (EEAR) breathing system and is characterised by the selective elimination of alveolar gas in all modes of ventilation. A functional description explains how this is achieved in the system in spontaneous and controlled ventilation. The potential for error which arises in combination systems for different switch positions with different modes of ventilation is avoided, as it is a single universal system. With no moving parts, mechanical reliability is guaranteed. The apparatus deadspace is negligible making the system usable even in neonates. Finally, the characteristics of the theoretical ideal non-absorber circuit are discussed as this illustrates the objective of the circuit design and important physical principles surrounding its function.

Anesthesia, Closed-Circuit↗
PubMed1995

Maxima and Bain breathing systems compared in controlled ventilation.

The Maxima is a new universal breathing system, which, despite its being valveless, functions with near maximal efficiency in spontaneous and controlled ventilation. It was compared to the Bain system in controlled ventilation by adjusting fresh gas flows (VF) to achieve an end-tidal CO2 of 5% in 40 patients aged from one to seventy-six years. They received a combined regional and general anaesthetic technique. The overall VF requirement of the Maxima system was: weight in kg x 35 ml.min-1.kg-1 + 1160 ml.min-1; the Bain system required VF of: weight in kg x 42 ml.min-1.kg-1 + 2070 ml.min-1. The mean VF requirement for the Bain system was greater by 45%. Of these forty patients, the results from the twenty-four adult patients weighing 50 kg or more gave the respective VF requirements for the Maxima and Bain systems of 52 and 71 ml.min-1.kg-1. In a separate study using twenty-nine patients, the fractional utilization of fresh gas (FU) (or VAe/VF where VAe = effective alveolar ventilation) in the Maxima and Bain systems was shown to be 0.94 and 0.71 respectively.

Adolescent↗
PubMed1995

Comparison in spontaneous ventilation of the Maxima with the Humphrey ADE breathing system and between four methods for detecting rebreathing.

An enclosed efferent, afferent reservoir breathing system (Maxima, Life Air Pty Ltd), being valveless, was compared to a simple afferent reservoir system (Humphrey ADE, A mode), having a valve, by assessing fresh gas flow (VF) requirements, with respect to ventilation (VE), that prevents rebreathing in volunteers, awake and breathing spontaneously. The results are recorded in terms of the quotient VF/VE associated with the onset of rebreathing. At the same time four clinical methods for assessing rebreathing were evaluated from the perspective of suitability for practical application during anaesthesia. No significant difference was found in the VF requirements between the ADE and Maxima breathing systems with respective values obtained for the quotient VF/VE of 0.80 and 0.77. Our findings showed that the method of sampling carbon dioxide (CO2) at the outflow of the efferent limb of the patient connector (eliminated CO2 method) in both afferent reservoir systems provided the highest flow rate and most reliable indication for detecting potential or actual rebreathing, when attempting to minimize VF. It provides no additional deadspace or resistance to gas flow, and has the advantage of being the only qualitative method that is reliable for the purpose of detecting the onset of rebreathing.

Airway Resistance↗
PubMed1995

The occurrence of mast cell tumors in young Shar-Peis.

During 1991 and 1992, 802 case submissions diagnosed as canine mast cell tumors were submitted to the Athens Veterinary Diagnostic Laboratory. Eighteen of these submissions were from dogs of the Shar-Pei breed. Of these 18 cases, 5 occurred in Shar-Peis less than 2 years of age, 4 of these were poorly differentiated, and 4 out of 5 occurred in the inguinal or preputial area. Three mast cell tumors (grade I) occurred in Boxer/Boxer cross dogs, and 2 tumors (grade II) occurred in Cocker Spaniels younger than 2 years of age. Of the 23,315 histopathology submissions, only 164 were from Shar-Peis and 57 of these dogs were under 2 years of age.

Animals↗
PubMed1995

Expression of the unc-4 homeoprotein in Caenorhabditis elegans motor neurons specifies presynaptic input.

In the nematode, Caenorhabditis elegans, VA and VB motor neurons arise from a common precursor cell but adopt different morphologies and synapse with separate sets of interneurons in the ventral nerve cord. A mutation that inactivates the unc-4 homeodomain gene causes VA motor neurons to assume the VB pattern of synaptic input while retaining normal axonal polarity and output; the disconnection of VA motor neurons from their usual presynaptic partners blocks backward locomotion. We show that expression of a functional unc-4-beta-galactosidase chimeric protein in VA motor neurons restores wild-type movement to an unc-4 mutant. We propose that unc-4 controls a differentiated characteristic of the VA motor neurons that distinguishes them from their VB sisters, thus dictating recognition by the appropriate interneurons. Our results show that synaptic choice can be controlled at the level of transcription in the post-synaptic neuron and identify a homeoprotein that defines a subset of cell-specific traits required for this choice.

Animals↗
PubMed1994

A practical approach to antibiotic treatment in women taking oral contraceptives.

Clinical situations that require the use of systemic antibiotic therapy are common. Because millions of women choose oral contraceptives for birth control, the potential for interaction between these drugs frequently has to be considered. We review the available information and present a practical approach for dealing with this situation based on sharing responsibility with an educated patient.

Adult↗
PubMed1994

The G-C specific DNA binding drug, mithramycin, selectively inhibits transcription of the C-MYC and C-HA-RAS genes in regenerating liver.

Expression of the c-myc and c-Ha-ras protooncogenes is dramatically increased in regenerating rat liver as an early response to partial hepatectomy. Nuclear runon transcription studies confirm that the increased c-myc and c-Ha-ras mRNA levels in regenerating livers reflect transcriptional activation of these genes. Mithramycin, a G-C specific DNA binding drug, prevents the increased transcriptional activity of c-myc and c-Ha-ras genes after hepatectomy but does not alter the transcriptional activity of the beta-actin gene. Continuous exposure of rats to mithramycin after hepatectomy prevents the increase in both c-myc and c-Ha-ras expression and blocks the increased cellular proliferation characteristic of regeneration. The delayed increase in c-myc and c-Ha-ras gene expression is associated with a delay in cellular proliferation. The inhibition of c-myc and c-Ha-ras transcription by mithramycin, the delay in cellular proliferation, and the ability of mithramycin to prevent protein binding to the c-myc promoter, suggest that the increased expression of these genes is a necessary component of liver regeneration.

Animals↗
PubMed1994

Mithramycin inhibits myointimal proliferation after balloon injury of the rat carotid artery in vivo.

BACKGROUND: Smooth muscle proliferation and extracellular matrix formation in the subintimal region of blood vessels that have been subjected to intimal injury are responsible for restenosis following balloon angioplasty of the coronary arteries and for accelerated atherosclerosis in a variety of other pathophysiological states. The immediate early-response gene c-myc is overexpressed in proliferating vascular smooth muscle cells in vitro, and c-myc antisense oligomers have been shown to reduce c-myc expression and to inhibit proliferation of vascular smooth muscle cells in culture. Mithramycin is a commercially available G-C-specific DNA binding drug that selectively inhibits transcription of genes, such as c-myc, that have G-C-rich promoter sequences. This study tested the hypothesis that mithramycin inhibits transcription of the c-myc proto-oncogene and prevents myointimal proliferation after balloon injury of the rat carotid artery in vivo. METHODS AND RESULTS: Ten-week-old male Sprague-Dawley rats received mithramycin (150 micrograms/kg IP) or distilled H2O 1 hour before and 1 hour after balloon injury of the right common carotid artery. After 2 weeks, the rats were killed by overdose of pentobarbital, and the injured right and uninjured control left arteries were pressure-fixed and subjected to morphological analysis for evaluation of the degree of myointimal thickening. Separate groups of rats were killed at 2 and 6 hours after vascular injury, and total RNA from injured and control vessels of mithramycin- and vehicle-treated rats was subjected to Northern analysis for assessment of steady-state c-myc mRNA levels. The areas of neointima and the ratios of neointimal to medial area were significantly less in mithramycin-treated than in control rats (0.6 +/- 0.1 versus 1.2 +/- 0.1 mm2, P < .01 and 95 +/- 16% versus 190 +/- 14%, P < .01). Lumen size was significantly greater in mithramycin-treated than in control rats (1.5 +/- 0.1 versus 0.8 +/- 0.1 mm2, P < .01). Steady-state c-myc mRNA levels were increased 10-fold and 2-fold (compared with undamaged carotid arteries) at 2 and 6 hours after balloon injury, respectively; mithramycin treatment reduced c-myc mRNA levels at 2 and 6 hours by 66% and 53%, respectively. CONCLUSIONS: These results support the hypothesis that systemic administration of mithramycin immediately (1 hour before and after intervention effectively inhibits transcription of the c-myc proto-oncogene and prevents myointimal proliferation after balloon injury of the rat carotid artery in vivo. Because mithramycin has been shown to be well tolerated by humans and to effectively inhibit transcription of c-myc in proliferating human cells, this agent may be useful in the prevention of coronary restenosis.

Angioplasty, Balloon↗
PubMed1994

The conjunctival wart: report of a case and review of treatment options.

Human papillomavirus, a papovavirus, is the causative agent of all warts. There are at least 63 subtypes of this virus, certain ones of which are more likely to cause warts in particular locations or to be associated with malignancies. Over the last 6 years, HPV-6, HPV-11, and HPV-16 have been detected in both benign and malignant lesions of the conjunctiva. We report a patient with a large viral papilloma of the palpebral conjunctiva of the right lower eyelid. This case demonstrates that even very large viral papillomas can be treated successfully.

Adult↗