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Biomedical subjects

K McEntee

Publications and source records attributed to K McEntee.

At least 19 recordsLinked to original sources
PubMed2003

Expression of the serotonin 1b receptor in experimental pulmonary hypertension.

The pathogenesis of pulmonary arterial hypertension (PAH) remains uncertain. Both the serotonin and endothelin (ET) systems are believed to be involved. Recent studies pointed to the importance of the serotonin 2B receptor as a limiting step. The current authors investigated the lung tissue expression of serotonin receptors and of the serotonin transporter (5-HTT) by real-time-quantitative polymerase chain reaction in chronic overcirculation-induced PAH in growing piglets, with and without treatment with the dual ET receptor blocker bosentan. Pulmonary haemodynamic changes were described by pulmonary arterial impedance spectra. Three months after the surgical anastomosis of the left subclavian artery to the pulmonary arterial trunk, there was a shift of the impedance spectra to higher ratios of pressure and flow moduli, with increases in both 0 Hz impedance and characteristic impedance, and these changes were completely prevented by bosentan therapy. There was an increase in the expression of the serotonin 1B receptor. There was no change in the expression of the 5-HTT, and of the serotonin 2B, 1D, and 4 receptors. The overexpression of the serotonin 1B receptor was partially prevented by bosentan therapy. The present authors conclude that this early pulmonary arterial hypertension model is characterised by an endothelin receptor-dependent increased expression of the serotonin 1B receptor.

Animals↗
PubMed2000

Use of ciliogenesis in the diagnosis of primary ciliary dyskinesia in a dog.

Primary ciliary dyskinesia is a congenital condition that may cause chronic rhinitis and bronchopneumonia. Primary ciliary dyskinesia may be diagnosed by induction of ciliogenesis by use of in vitro cell culture. Induction of ciliogenesis allows for differentiation between primary and secondary ciliary dyskinesia.

Animals↗
PubMed2000

Genetic analysis of the influence of pertussis toxin on experimental allergic encephalomyelitis susceptibility: an environmental agent can override genetic checkpoints.

Pertussis toxin (PTX) is a potent ancillary adjuvant used to elicit several different autoimmune diseases, including experimental allergic encephalomyelitis (EAE). To delineate the genetics of PTX effect in EAE, we mapped EAE-modifying (eae-m) loci in cohorts of backcross mice immunized with and without PTX. In this study, we analyzed the genetic basis of EAE susceptibility and severity and the intermediate phenotypes of mononuclear cell infiltration, suppuration, and demyelination. In animals immunized with PTX, one major locus, eae9, controls disease susceptibility and severity. Eae9 also regulates the extent of mononuclear cell infiltration of the spinal cord in male mice. Without PTX, five eae-m loci were noted, including three new loci in intervals on chromosomes 8 (eae14), 10 (eae17), and 18 (eae18). Taken together, these results suggest that eae9 controls the effects of PTX in EAE susceptibility, and is capable of overriding the other genetic checkpoints in the pathogenesis of this disease.

Animals↗
PubMed1999

Multiple independent regulatory pathways control UBI4 expression after heat shock in Saccharomyces cerevisiae.

Transcription of the polyubiquitin gene UBI4 of Saccharomyces cerevisiae is strongly induced by a variety of environmental stresses, such as heat shock, nutrient depletion and exposure to DNA-damaging agents. This transcriptional response of UBI4 is likely to be the primary mechanism for increasing the pool of ubiquitin for degradation of stress-damaged proteins. Deletion and promoter fusion studies of the 5' regulatory sequences indicated that two different elements, heat shock elements (HSEs) and stress response element (STREs), contributed independently to heat shock regulation of the UBI4 gene. In the absence of HSEs, STRE sequences localized to the intervals -264 to -238 and -215 to -183 were needed for stress control of transcription after heat shock. Site-directed mutagenesis of the STRE (AG4) at -252 to -248 abolished heat shock induction of UBI4 transcription. Northern analysis demonstrated that cells containing either a temperature-sensitive HSF or non-functional Msn2p/Msn4p transcription factors induced high levels of UBI4 transcripts after heat shock. In cells deficient in both heat stress pathways, heat-induced UBI4 transcript levels were considerably lower but not abolished, suggesting a role for another factor(s) in stress control of its expression.

Actins↗
PubMed1999

Doppler echocardiographic study of left and right ventricular function during dobutamine stress testing in conscious healthy dogs.

OBJECTIVE: To evaluate left and right ventricular filling and ejection performances by use of Doppler echocardiography in healthy, conscious dogs submitted to dobutamine stress testing. ANIMALS: 10 unsedated, healthy adult Beagles. PROCEDURE: Doppler echocardiography was performed during cardiac stress testing on each dog twice at 24-hour intervals. Dobutamine was infused in 10 micrograms/kg of body weight/min incremental dosages, from 12.5 to 42.5 micrograms/kg/min. Duration of each step was 15 minutes. Doppler measurements were recorded at baseline and at each stage of dobutamine infusion, whereas aortic diameter was measured at baseline and at peak dosage by use of two-dimensional echocardiography. RESULTS: Dobutamine infusion induced a significant increase in velocity time integrals and in peak flow velocities at the aortic, pulmonic, mitral, and tricuspid valves. Acceleration time-to-deceleration time ratio at the aortic wave also was increased significantly. On the other hand, ejection time, acceleration time, and deceleration time at the aortic and pulmonic valves and peak flow velocity of the E wave-to-peak flow velocity of the A wave ratio at the mitral and tricuspid valves decreased significantly during the test. The acceleration time-to-deceleration time ratio at the pulmonic wave was unchanged. A significant, progressive increase in cardiac index also was observed during dobutamine infusion, with a maximal increase of 104% from baseline. This was mediated initially by an increase in stroke index and, at higher dosages, by an increase in heart rate. CONCLUSIONS AND CLINICAL RELEVANCE: Doppler echocardiography performed during dobutamine stress testing may be a reliable method of assessing myocardial function in dogs with cardiovascular disease.

Animals↗
PubMed1998

Transcriptional factor mutations reveal regulatory complexities of heat shock and newly identified stress genes in Saccharomyces cerevisiae.

A computer-aided pattern search of the entire yeast genome was designed and used to identify 186 putative stress response element-regulated genes in Saccharomyces cerevisiae. Transcript levels of eight of these candidate genes were examined, and three (37%) were shown to be heat shock- and DNA damage-inducible and to require the Msn2p and Msn4p transcriptional activators for stress regulation. Significantly, several heat shock protein (HSP) genes were identified in this computer search. Using a series of single and multiple regulatory mutants, we demonstrate unexpected regulatory complexities among the HSP genes from S. cerevisiae following heat shock.

Databases, Factual↗
PubMed1998

Laryngeal rhabdomyoma in a golden retriever.

A three-year-old male golden retriever had had progressive dyspnoea, exercise intolerance, stridor, and a modified bark for five months. A mass 2 cm in diameter was present dorsal to the right side of the larynx. Histological examination revealed cross-striations in some elongated cells, consistent with a diagnosis of rhabdomyoma, a diagnosis which was confirmed by positive immunohistochemical staining for myoglobin and desmin. The mass could not be removed without total laryngectomy and a permanent tracheostomy and the dog was euthanased.

Animals↗
PubMed1998

Functional analysis of the stress response element and its role in the multistress response of Saccharomyces cerevisiae.

The DDR2 gene of Saccharomyces cerevisiae is a multistress response gene whose transcription is rapidly and strongly induced by a diverse array of xenobiotic agents, and environmental and physiological conditions. The multistress response of this gene requires the pentanucleotide, 5' CCCCT, (C4T;STRE (STress Response Element)) and the zinc-finger transcription factors, Msn2p and Msn4p. A 51bp oligonucleotide (oligo 31/32) containing two STREs from the DDR2 promoter region was previously shown to direct heat shock activation of a lacZ reporter gene. In this work we demonstrate that the same element conferred a complete multistress response to an E. coli galK reporter gene introduced into yeast cells. A variant oligonucleotide in which both the STRE spacing and neighboring sequences were altered responded to the same spectrum of stresses, while substitution of nucleotides within the pentanucleotide completely abolished the multistress response. These results directly demonstrate that STREs are not only necessary but are sufficient for mediating a transcriptional response to a surprisingly diverse set of environmental and physiological conditions.

Base Sequence↗
PubMed1998

Physiologic response to dobutamine infusion during cardiac stress testing of dogs.

OBJECTIVE: To evaluate response of various cardiovascular variables after administration of incremental doses of dobutamine in healthy conscious dogs, using standardized dobutamine stress echocardiography (DSE). ANIMALS: 8 healthy dogs. PROCEDURE: A DSE was performed twice on each dog within 24 hours. Dobutamine was infused at a rate of 12.5 to 42.5 microg/kg/min, using incremental increases of 10 microg/kg/min. Doppler sphygmomanometry, electrocardiography, and echocardiography were performed. Left ventricular size, global ventricular performance, and left ventricular systolic myocardial function were measured by means of echocardiography. RESULTS: At the highest dosage, dobutamine induced an increase of 20+/-3% and 109+/-12% in systolic blood pressure and cardiac index, respectively. The latter was associated with a significant increase in heart rate and stroke index. Fractional shortening of the left ventricle, fractional thickening of the left ventricular free wall and interventricular septum, ejection fraction, and mean velocity of fiber shortening had a progressive and significant increase during dobutamine infusion. Preejection period and left ventricular ejection time had a progressive and significative decrease during the stress test. CONCLUSIONS: The technique used was feasable, safe, and repeatable in healthy conscious dogs. Control values were determined. CLINICAL RELEVANCE: Data for these healthy dogs might be useful for comparison with results obtained from dogs with known or suspected cardiovascular disease.

Animals↗
PubMed1996

Structure and functional analysis of the multistress response gene DDR2 from Saccharomyces cerevisiae.

The DDR2 gene is a multistress response gene in Saccharomyces cerevisiae that is transcriptionally activated by more than thirteen xenobiotic agents and environmental or physiological stresses. The DDR2 gene encodes a small hydrophobic 61 amino acid polypeptide located on chromosome XV adjacent to the SPE2 locus. Disruption alleles of the DDR2 gene have been constructed and these ddr2 delta mutants show no defect in heat shock recovery or thermotolerance and appear normal for DNA damage sensitivity and mutagenesis.

Amino Acid Sequence↗
PubMed1996

Msn2p, a zinc finger DNA-binding protein, is the transcriptional activator of the multistress response in Saccharomyces cerevisiae.

The stress response promoter element (STRE) confers increased transcription to a set of genes following environmental or metabolic stress in Saccharomyces cerevisiae. A lambda gt11 library was screened to isolate clones encoding STRE-binding proteins, and one such gene was identified as MSN2, which encoded a zinc-finger transcriptional activator. Disruption of the MSN2 gene abolished an STRE-binding activity in crude extracts as judged by both gel mobility-shift and Southwestern blot experiments, and overexpression of MSN2 intensified this binding activity. Northern blot analysis demonstrated that for the known or suspected STRE-regulated genes DDR2, CTT1, HSP12, and TPS2, transcript induction was impaired following heat shock or DNA damage treatment in the msn2-disrupted strain and was constitutively activated in a strain overexpressing MSN2. Furthermore, heat shock induction of a STRE-driven reporter gene was reduced more than 6-fold in the msn2 strain relative to wild-type cells. Taken together, these data indicate that Msn2p is the transcription factor that activates STRE-regulated genes in response to stress. Whereas nearly 85% of STRE-mediated heat shock induction was MSN2 dependent, there was significant MSN2-independent expression. We present evidence that the MSN2 homolog, MSN4, can partially replace MSN2 for transcriptional activation following stress. Moreover, our data provides evidence for the involvement of additional transcription factors in the yeast multistress response.

Base Sequence↗
PubMed1996

Purification of a soluble UmuD'C complex from Escherichia coli. Cooperative binding of UmuD'C to single-stranded DNA.

The Escherichia coli UmuD' and UmuC proteins play essential roles in SOS-induced mutagenesis. Previous studies investigating the molecular mechanisms of mutagenesis have been hindered by the lack of availability of a soluble UmuC protein. We report the extensive purification of a soluble UmuD'C complex and its interactions with DNA. The molecular mass of the complex is estimated to be 70 kDa, suggesting that the complex consists of one UmuC (46 kDa) and two UmuD' (12 kDa) molecules. In contrast to its inability to bind to double-stranded DNA, UmuD'C binds cooperatively to single-stranded DNA as measured by agarose gel electrophoresis and confirmed by steady-state fluorescence depolarization. A Hill coefficient, n = 3, characterizes the binding of UmuD'C to M13 DNA and to a 600 nucleotide DNA oligomer, suggesting that at least three protein complexes may interact cooperatively when binding to DNA. The apparent equilibrium binding constant of UmuD'C to single-stranded DNA is approximately 300 nM. Binding of the complex to a short, 80 nucleotide, DNA oligonucleotide was detectable by fluorescence depolarization, but it did not appear to be cooperative. Binding of UmuD'C to single-stranded M13 DNA causes an acceleration of the protein-DNA complex, suggesting that the longer DNA may undergo compaction. The UmuD'C complex associates with RecA-coated DNA, and the UmuD'C complex remains bound to DNA in the presence of RecA.

Amino Acid Sequence↗
PubMed1996

Cardiac performance in conscious healthy dogs during dobutamine infusion.

The aim of this study was to determine the cardiac performance of conscious healthy dogs during stimulation with dobutamine. Eight healthy unsedated beagle dogs were used. Cardiac output was measured by the thermodilution technique and blood pressures by extravascular pressure transducers. Dobutamine challenge at a dosage ranging from 27.5 to 50 micrograms kg-1 min-1 induced a significant rise in cardiac power index (CPI), cardiac index (CI), stroke index (SI) and heart rate (HR) and a significant decrease in pulmonary vascular resistance (PVR) and systemic vascular resistance (SVR). The highest CPI was 2.05 times greater than its basal resting value. The CI was primarily responsible for this increase in CPI. The SI and HR contributed approximately 55 per cent and 45 per cent respectively of the maximal increase in CI.

Analysis of Variance↗
PubMed1995

Purification and properties of wild-type and exonuclease-deficient DNA polymerase II from Escherichia coli.

Wild-type DNA polymerase II (pol II) and an exonuclease-deficient pol II mutant (D155A/E157A) have been overexpressed and purified in high yield from Escherichia coli. Wild-type pol II exhibits a high proofreading 3'-exonuclease to polymerase ratio, similar in magnitude to that observed for bacteriophage T4 DNA polymerase. While copying a 250-nucleotide region of the lacZ alpha gene, the fidelity of wild-type pol II is high, with error rates for single-base substitution and frameshift errors being < or = 10(-6). In contrast, the pol II exonuclease-deficient mutant generated a variety of base substitution and single base frameshift errors, as well as deletions between both perfect and imperfect directly repeated sequences separated by a few to hundreds of nucleotides. Error rates for the pol II exonuclease-deficient mutant were from > or = 13- to > or = 240-fold higher than for wild-type pol II, depending on the type of error considered. These data suggest that from 90 to > 99% of base substitutions, frameshifts, and large deletions are efficiently proofread by the enzyme. The results of these experiments together with recent in vivo studies suggest an important role for pol II in the fidelity of DNA synthesis in cells.

Base Sequence↗
PubMed1995

Aod1, the immunoregulatory locus controlling abrogation of tolerance in neonatal thymectomy-induced autoimmune ovarian dysgenesis, maps to mouse chromosome 16.

Mice thymectomized at three days of age (D3Tx) develop during adulthood a variety of organ-specific autoimmune diseases, including autoimmune ovarian dysgenesis (AOD). The phenotypic spectrum of AOD is characterized by the development of anti-ovarian autoantibodies, oophoritis, and atrophy. The D3Tx model of AOD is unique in that disease induction depends exclusively on perturbation of the normal developing immune system, is T-cell-mediated, and is strain specific. For example, D3Tx A/J mice are highly susceptible to AOD, whereas C57BL/6J mice are resistant. After D3Tx, self ovarian antigens, expressed at physiological levels, trigger an autoimmune response capable of eliciting disease. The D3Tx model provides, therefore, the opportunity to focus on the mechanisms of self-tolerance that are relevant to disease pathogenesis. Previous studies indicate that the principal mechanisms involved in AOD susceptibility are genetically controlled and govern developmental processes associated with the induction and maintenance of peripheral tolerance. We report here the mapping of the Aod1 locus to mouse chromosome 16 within a region encoding several loci of immunologic relevance, including scid, Igl1, VpreB, Igll, Igl1r, Mtv6 (Mls-3), Ly-7, Ifnar, and Ifgt.

Animals↗
PubMed1995

Closantel intoxication in a dog.

A case of overdosage with closantel, a salicynalide derivative, in a dog is described. The dog received 6 times the recommended dosage. Closantel induced optic neuritis, retinal degeneration, partial deafness, hepatotoxicosis and myopathy. Only the blindness was irreversible. The therapy included albumin administration to reduce the acute toxicity of closantel.

Albumins↗
PubMed1995

Comparative assessment of right ventricular performance from the pressure-volume relationship in double-muscled and conventional calves.

Forty-one and 55 records of right-sided and systemic arterial pressures, cardiac output, and end-diastolic and end-systolic right ventricular volumes were collected from a group of 6 conventional and 6 double-muscled calves, respectively. In each group, the mean right ventricular pressure-volume loop was constructed. Global cardiac performance was significantly lower in the double-muscled than in the conventional calves. The right ventricular end-diastolic and end-systolic volumes, as well as the diastolic portion of the mean pressure-volume loop, were similar in the 2 groups. Those results suggest that the reduced cardiac performance of double-muscled calves is not due to a lowered ventricular preload and that diastolic properties of their myocardium are similar to those of conventional calves. When expressed on a body weight basis, however, the right ventricular end-diastolic and end-systolic volumes were lower in the double-muscled than in conventional calves. When expressed as a function of probable metabolic demand, therefore, the volumetric capacity of the cardiac pump appears to be reduced in double-muscled calves. The significantly lower right ventricular ejection fraction, maximal rate of ventricular pressure rise and right ventricular peak-systolic pressure to end-systolic volume ratio measured in double-muscled as compared with conventional calves suggest that reduced myocardial contractility may also be partly responsible for the significantly lower stroke index of the former calves. The cardiac pump of double-muscled cattle thus seems to be less effective than that of conventional cattle because of reduced volumetric capacity and lowered strength of contraction.

Animals↗