Biomedical subjects

J Burnside

Publications and source records attributed to J Burnside.

At least 37 records · Page 2Linked to original sources

PubMed1997

Molecular cloning and sequence analysis of chicken type I deiodinase cDNA: expression in normal and dwarf broiler chickens.

A cDNA encoding the chicken type I iodothyronine deiodinase (cDI-1) was isolated and sequenced from a cDNA library prepared from ConA-activated chicken splenic T-lymphocytes. The coding region of cDI-1 cDNA is composed of 738 basepairs (bp) which encodes a 246 amino acid protein. The predicted amino acid sequence of cDI-1 indicates only 60% identity to several mammalian type I deiodinases. The cDI-1 cDNA contains a codon for a highly conserved selenocysteine residue (Cys124). Northern blot analysis of total RNA prepared from different tissues of a 3-week-old broiler chicken shows a single transcript (2 kb) in liver and kidney. The abundance of hepatic cDI-1 transcripts in growth hormone receptor (GHR)-deficient dwarf chicken was similar to normal chickens despite lower levels of plasma T3 (37% lower) and elevated levels of T4 (21% higher) in dwarf chickens. This finding suggests that regulation of hepatic cDI-1 mRNA is GH-independent in the post-hatch chicken.

Amino Acid Sequence↗

PubMed1997

Growth hormone down-regulates growth hormone receptor mRNA in chickens but developmental increases in growth hormone receptor mRNA occur independently of growth hormone action.

The purpose of this study was to determine the role of growth hormone (GH) in regulating expression of the chicken GH receptor (cGHR) gene by comparing the levels of cGHR mRNA in livers of normal chickens with that of GHR-deficient dwarf chickens. Since the sex-linked dwarf chicken lacks a functional cGHR, there are no genes activated as a result of GH action. Examination of the early developmental profile of hepatic cGHR mRNA in normal and dwarf chickens should yield information on the relative contribution of developmental and hormonal factors to the regulation of cGHR gene expression. Using a sensitive RNase protection assay, we found that the abundance of the major cGHR transcripts (4.3, 3.2 and 0.8 kb) in normal chickens increases about 2-fold between 1 and 7 weeks of age. Due to a splice site mutation in the dwarf chicken, the two larger transcripts encoding the full-length cGHR are not expressed. However, the expression of the truncated cGHR transcript (0.8 kb) in dwarf chickens increases about 5-fold between 1 and 7 weeks of age which suggests that the cGHR gene is overexpressed when not down-regulated by GH. Furthermore, a single promoter, appears to control expression of cGHR transcripts in liver since primer extension analysis revealed the same 5'-end in both full-length and 0.8 kb transcripts. These observations suggest that even though developmental increases in cGHR gene expression occur independently of GH action, GH, either directly or indirectly, down-regulates expression of the cGHR gene in normal chickens.

Animals↗

PubMed1997

Chronic administration of growth hormone (GH) to adult chickens exerts marked effects on circulating concentrations of insulin-like growth factor-I (IGF-I), IGF binding proteins, hepatic GH regulated gene I, and hepatic GH receptor mRNA.

In young birds, growth hormone (GH) administration has been found to have only a small or even no effect on circulating concentrations of insulin-like growth factor-I (IGF-I). This is in obvious contrast to the situation in mammals. The present study examines the effect of continuous administration of GH in adult male chickens. Plasma concentrations of IGF-I were markedly elevated (2.5-3.0-fold, p < 0.001) in GH-treated chickens. There were also some transient increases in the circulating levels of IGF binding proteins. Adult chickens showed other manifestations of increased responsiveness to GH, including elevated hepatic expression of GH-regulated gene-I (mRNA) with GH treatment (p < 0.05), and a tendency (p < 0.08) for decreased GH-receptor mRNA. In contrast to the changes in circulating concentrations of GH and IGF-I with GH treatment, no changes in plasma concentrations of thyroid hormones, reproductive hormones, glucose, or nonesterified fatty acids were evident.

Animals↗

PubMed1996

Interferon activity of mitogen-induced chicken splenic lymphocytes which do not express interferon mRNA.

Interferon activity was measured in media from virally infected chicken embryo fibroblasts and Concanavalin A-stimulated splenic lymphocytes using a viral inhibition assay. Both cell types produce interferon activity. A cDNA probe corresponding to a chicken interferon mRNA was used to probe Northern blots of RNA prepared from both cells. A single hybridizing species of 900 bases was detected in virally infected fibroblast RNA, but no hybridizing species was detected in the splenic lymphocytes. These results suggest that the interferon activity produced by lymphocytes is of different molecular origin than the corresponding activity produced by virally infected fibroblasts.

Animals↗

PubMed1995

Comparison of gene expression in normal and growth hormone receptor-deficient dwarf chickens reveals a novel growth hormone regulated gene.

Because of a dysfunctional growth hormone (GH) receptor there is an absence of GH-dependent gene expression in the sex-linked dwarf chicken. Therefore, a comparison of mRNAs expressed in normal and dwarf chickens should lead to the identification of mRNAs that are regulated by GH action. We have compared gene expression in normal and dwarf chickens using the mRNA differential display technique. A combination of three anchored oligo dT primers and 15 random decamers were used to detect at least 75 differentially expressed mRNAs. One of these, designated GHRG-1, hybridizes to a 0.9 kb transcript found only in liver and in normal chickens shows a pattern of developmental expression which parallels the plasma GH profile. A GHRG-1 cDNA clone was isolated that encodes a 120 amino acid peptide with no homology to any known gene. Sequence of the promoter from a genomic clone shows a region with strong similarity to the GH response element identified in the serine protease inhibitor gene, Spi 2.1. These results suggest that GHRG-1 is a novel GH regulated gene.

Aging↗

PubMed1995

The Australian Capital Territory Injury Surveillance and Prevention Project.

OBJECTIVE: To identify the representativeness and sensitivity of the data collected by the Australian Capital Territory Injury Surveillance and Prevention Project (ACTISPP) for ED injury cases seen in the Australian Capital Territory (ACT). METHODS: A retrospective descriptive study was conducted using a systematic sample of people injured in the ACT between January 20 and September 19, 1992, and who received their first treatment for this injury in an ACT ED. The ACTISPP surveillance system is dependent on ED patient or family completion and return of a comprehensive injury survey. The following population variables in the ACTISPP database were compared with those of the injured ED population, as determined by ED and hospital record review: age, gender, postal code of residence, nature of injury, body part injured, and whether the patient had been admitted to hospital. RESULTS: The sensitivity of the surveillance system for injuries seen in the ED was 31% (95% CI = 30-32%). The distribution of the ACTISPP sample did not differ from the expected distribution for sex or socioeconomic status. The distributions differed by age only in the 5-14-year age group (p < 0.001). With respect to the nature of injury and body part injured, the difference was significant (p < 0.001) but was no greater than 7% for any of the categories. Two percent of the surveillance sample were admitted to hospital, compared with an estimated 4% of the target population (p < 0.001). CONCLUSIONS: Approximately one-third of all incident injuries managed in the EDs of the ACT are included in the ACTISPP database. Although some sampling bias is introduced in the surveillance process, it does not appear to be of sufficient magnitude to compromise the project's goals.

Adolescent↗

PubMed1995

Accuracy of in-line venous saturation and hematocrit monitors in pediatric perfusion.

Cardiopulmonary bypass (CPB) in neonates and infants routinely employs lower blood flow rates (BFR), temperatures, and hematocrits (Hct) than those typically observed in adult CPB. The purpose of this study was to evaluate the accuracy of three devices available for continuous in-line measurement of venous oxygen saturation (SvO2) and Hct during pediatric CPB. Venous blood samples were obtained over a range of BFR, temperatures, and Hct and analyzed on a Corning 2500 Co-Oximeter and HematoStat C-70 centrifuge. These values were then compared to those measured by the Bentley OxySat SM-0200, the Gish StatSat, and the 3M CDI 100 in-line monitors. Seventy samples were obtained and analyzed using linear regression, paired t-test and residual analysis to establish the reliability and accuracy of each device. The results demonstrate the CDI to be more statistically accurate (p > 0.05) than the Gish and Bentley devices for measuring SvO2 in nearly all circumstances, though all correlated well with the control (r > 0.70). When comparing spun Hct to the CDI and Gish values, the CDI in-line monitor demonstrated a greater reliability to predict actual patient Hct (r > 0.90) than the Gish StatSat (r > 0.60). Residual analysis revealed that even though the Gish StatSat had higher calculated p values (p > 0.05) than the CDI 100 for interpreting Hct, it was shown to display more inconsistent and sporadic values over the ranges of BFR and temperature studied. It is concluded that the CDI 100 proved to be more accurate, reliable, and consistent than the Gish StatSat and the Bentley Oxysat devices in determining SvO2 and Hct over all evaluated parameters in this study.

Adult↗

PubMed1994

Forearm compartment syndrome following thrombolytic therapy for acute myocardial infarction.

Thrombolytic therapy for myocardial infarction may contribute to bleeding complications when central venous or arterial access is required, but peripheral venous access is usually uncomplicated. We report a patient in whom tissue plasminogen activator and subsequent intravenous heparin exacerbated bleeding from a disrupted intravenous access site, leading to acute compartment syndrome requiring surgical decompression. This case emphasizes the risks associated with iatrogenic trauma during thrombolytic therapy.

Compartment Syndromes↗

PubMed1994

Dysfunctional growth hormone receptor in a strain of sex-linked dwarf chicken: evidence for a mutation in the intracellular domain.

The sex-linked dwarf (dwdw) chicken represents a valuable animal model for studying GH insensitivity and the consequence of mutations in the GH receptor (GHR) gene. We have recently reported undetectable hepatic GH-binding activity and an aberrantly sized transcript in a strain of dwdw chickens obtained from Arbor Acre Farms, Inc. (Glastonbury, CT, USA). Southern blot analysis of the chicken GHR (cGHR) gene revealed a restriction-fragment length polymorphism in HindIII and EcoRI digests of genomic DNA in this strain of dwdw chicken. In order to localize the molecular mutation, we analysed the gene structure and determined the complete sequence of the 3' untranslated region (3' UTR) of the normal cGHR. With the use of this information, we located a large deletion in the 3' end of the cGHR gene of the Connecticut (CT) strain of dwdw chicken. This deletion (1773 bp) contained 27 highly conserved amino acids of the 3' end of the coding region, the in-frame stop codon, a less frequently used poly(A) signal that is normally found 445 bp downstream of the stop codon, and a large portion of the 3' UTR. Because of this deletion, 27 novel amino acids were substituted and the open reading frame was extended for an additional 26 amino acids before reaching the transcriptional termination site. The predicted amino acid sequence of the novel carboxyl-terminus of the dwdw cGHR is largely hydrophobic with a polylysine tail, whereas the carboxyl-terminus of the wild-type (DwDw) cGHR is composed of hydrophilic amino acids.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

PubMed1993

Thyroid hormone regulation of thyrotropin gene expression.

Thyroid hormones suppress the synthesis and release of thyrotropin from thyrotropes in the anterior pituitary gland, a feature that is critical in the classic negative-feedback loop of the pituitary-thyroid endocrine axis. The major effect of thyroid hormones in this system is exerted at the transcriptional level. The molecular mechanisms by which there is negative regulation of TSH subunit gene expression by thyroid hormone have been elucidated. The TSH subunit genes have isolated and characterized. Structure-function analyses using fusion genes and DNA transfection approaches have defined the putative negative TREs among the promoters of the rat, mouse, and human alpha and TSH beta genes. These sequences are either largely overlapping direct TRE half-sites, TRE half-sites as direct repeats gapped by two nucleotides, or single TRE half-sites. These arrangements are distinct from those seen in positive TREs. Recent knowledge regarding the molecular mechanisms of thyroid action in general forces consideration of multiple TR isoforms, TR heterodimer partners (TRAPs), and thyroid hormones in the ultimate mechanisms of negative action. Several models have been proposed, but none has yet been proved. In addition, the role of thyroid hormone in the regulation of gene expression at the posttranscriptional level is beginning to be addressed. Future work should continue to illuminate these important facets of gene regulation.

Animals↗

PubMed1993

Overexpression of a truncated growth hormone receptor in the sex-linked dwarf chicken: evidence for a splice mutation.

Sex-linked dwarfism in chickens is a form of GH resistance that resembles the Laron syndrome in humans. The dwarfism found in chickens is due to a mutant gene (dw) carried on the sex chromosome. The homozygous dwarf (dwdw) chicken is characterized by reductions in stature and plasma insulin-like growth factor-I (IGF-I) levels. Despite the absence of hepatic GH-binding activity, Southern blot analysis shows that there is no gross structural change in the gene for the GH receptor (GHR) in this strain of dwdw chicken. GH-dependent IGF-I production can be restored in cultured dwdw hepatocytes after transfection and transient expression of a chicken GHR (cGHR) cDNA, indicating that other factors that participate in GH-mediated IGF-I synthesis are intact. Northern blot analysis of liver, muscle, fat, and pituitary RNA from normal (DwDw) chickens shows a major transcript of 4.3 kilobases (kb) and three minor transcripts (0.8, 1.7, and 3.2 kb), which correspond to the cGHR. In contrast, the 0.8-kb transcript is the major cGHR transcript expressed in these tissues from dwdw chickens. Northern blot analysis with domain-specific probes shows that the 0.8-kb transcript in DwDw and dwdw liver contains only a small portion of the extracellular domain of the cGHR. A cDNA clone encoding this transcript has been isolated from a liver library prepared from a normal chicken.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

PubMed1993

Clinical comparisons of continuous venous oxygen saturation and hematocrit monitors in pediatric surgery.

Continuous venous oxygen saturation and hematocrit values are important parameters in assessing patient status while on cardiopulmonary bypass. Two devices used to measure continuous venous oxygen saturation while on cardiopulmonary bypass were compared to a control. The Bentley Oxysat meter and the Medtronic MX2 Oxygen Saturation and Hematocrit System were compared to the ABL500 blood gas monitor. The continuous hematocrit readings from the MX2 system were compared to spun hematocrits. Twenty-nine pediatric patients ranging from 2.3kg-43.3kg were randomly selected. In-line optical transmission cells used were 1/4" or 3/8" depending upon the patient's blood flow requirements. A total of 163 data points were analyzed at different blood flow rates, temperatures, and hematocrits. The venous oxygen saturation values obtained from the Oxysat and the MX2 devices correlated well with the ABL500 over the entire range of blood flows, temperatures, and hematocrits. All correlation coefficients were greater than 0.89. The correlation between the MX2 device and the spun hematocrit varied with temperature ranges. The correlation coefficient tended to decrease with decreasing temperature. We conclude that these devices are important adjuncts to bypass as long as their limitations are understood.

Blood Gas Analysis↗

PubMed1992

Abnormal growth hormone receptor gene expression in the sex-linked dwarf chicken.

Sex-linked dwarfism is a recessive mutation that causes a reduction in body weight gain and long bone growth of chickens. We examined the effect of the dwarfing gene on body weight, hepatic GH-binding activity, and the structure and expression of the growth hormone receptor (GHR) gene in two different lines of sex-linked dwarf (SLD) broiler chickens. Liver samples from one line of dwarf chicken were obtained from Arbor Acres Farm, Inc. (Glastonbury, CT) and fertile eggs from the second line of SLD were obtained from the University of Georgia. In the GA line, the average body weight of homozygous (dwdw) males at 11 weeks of age was 43% lower than that of normal (DwDw) males, while heterozygous (Dwdw) males were only 9% below normal. In the CT line, hepatic GH-binding activity of 35-week-old chickens was high (20% specific binding) in normal (DwDw) males and undetectable in liver membranes prepared from dwdw males. At 11 weeks of age, hepatic GH-binding activity of Dwdw males (3.9% specific binding) in the GA line was reduced by 44% and that of dwdw males was almost undetectable (0.34% specific binding) when compared to the average of normal GA males (7.1% specific binding). Southern and Northern blot analyses revealed different abnormalities in the GHR gene from the two separate lines of SLD. A restriction fragment length polymorphism in DNA and an aberrantly sized transcript (mRNA) were detected in the CT line of SLD chickens.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

Developmental expression of hepatic growth hormone receptor and insulin-like growth factor-I mRNA in the chicken.

We have examined the ontogeny of expression of growth hormone (GH) receptor (GHR) and insulin-like growth factor-I (IGF-I) mRNA in chicken liver from day 13 of incubation until 31 weeks of age. The profiles of GHR and IGF-I mRNA levels were compared to developmental changes in body weight and plasma levels of GH and IGF-I. In the embryo, hepatic GHR mRNA was not detectable until day 15, highest on days 17 and 19, and then declined at hatching (day 21). Following an initial 2-week delay after hatching, there was a progressive increase in hepatic GHR mRNA which continued after the birds reached mature body weight. Plasma GH reached peak levels at 3-4 weeks of age and then fell sharply until maintenance of a low basal level after 10 weeks of age. Thus, there appears to be a strong inverse relationship between expression of the GHR and basal plasma GH levels in the prepubertal chicken. Although IGF-I mRNA was undetectable in embryonic liver by Northern blot analysis, there is a good correlation between expression of hepatic IGF-I mRNA and the plasma IGF-I profile during post-hatching development in the chicken. The highest levels of IGF-I mRNA were reached at 4 weeks of age which was followed by a slow decline to the basal levels maintained after 10 weeks of age. It appears that the decline in plasma IGF-I lags considerably behind the sharp fall in plasma GH levels and expression of hepatic IGF-I mRNA.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1991

Molecular cloning of the chicken growth hormone receptor complementary deoxyribonucleic acid: mutation of the gene in sex-linked dwarf chickens.

A novel complementary DNA (cDNA) encoding the chicken GH receptor was isolated from a chicken liver cDNA library, using polymerase chain reaction with primers derived from highly conserved sequences of the mammalian GH receptor. The nucleotide sequence predicts a mature protein of 592 amino acids and a 16 amino acid signal peptide that are partially homologous to the sequence reported for the rabbit (53%), rat (58%), and human (50%) GH receptors. Despite this low level of homology, a number of structural features of the GH receptor are conserved, including 7 cysteine residues in the extracellular domain and 5 in the intracellular region. Three transcripts of approximately 4.7, 4.0, and 1.0 kilobases are present on Northern blots of total RNA prepared from the livers of 35-week-old male chickens. Expression of the GH receptor was also detected in a wide range of tissues. The chicken GH receptor cDNA was then used as a probe in Southern and Northern blot analyses of DNA and RNA prepared from livers of sex-linked dwarf chickens, which have undetectable levels of hepatic GH-binding activity, in addition to other endocrine abnormalities. A restriction fragment length polymorphism was found in DNA, and an aberrantly-sized transcript was found in hepatic RNA of the dwarf chicken. These results indicate that a mutation in the GH receptor gene is responsible for the phenotype of the sex-linked dwarf chicken. This type of dwarfism resembles Laron-type dwarfism in humans, where a defect in the GH receptor gene has recently been identified. These receptor-deficient chickens should serve as a unique model system for studying the role of the GH receptor in growth and development.

Amino Acid Sequence↗

PubMed1991

3,5,3'-triiodothyronine (T3) receptor-auxiliary protein (TRAP) binds DNA and forms heterodimers with the T3 receptor.

Nuclear factors enhance binding of T3 receptors (TR) to DNA, suggesting that T3 action may require a multicomponent complex bound to thyroid hormone response elements (TREs). We refer to the 65,000 Da nuclear protein in GH3 cells that enhances TR binding to DNA as the TR-auxiliary protein (TRAP) and have characterized its interaction with TR. Using a TRE-DNA affinity matrix we show that TRAP is able to bind to DNA, even in the absence of functional TR. We then used carboxyl-terminal truncations of rat TR alpha-1 and human TR beta in the avidin-biotin complex DNA-binding assay to identify regions that are important for interaction with TRAP. Removal of 34 residues of hTR beta abolishes T3-binding activity, but the ability to bind TRAP is retained. Further truncations and point mutations suggest that TRAP interacts with the ligand-binding domain of TR and with an independent region which overlaps a conserved sequence adjacent to the second Zn2+ finger (amino acids 120-149 in rTR alpha-1). A fragment of rTR alpha-1 (alpha C291) which encompasses these two regions inhibits the ability of TRAP to enhance TR binding to DNA. This is due to binding of alpha C291 to TR, demonstrating the ability of TR to form homodimers. The inability of TRAP to interact with TR dimers and the similarity of the locations of the estradiol receptor dimerization domains with the TRAP interaction regions lead us to conclude that TRAP stabilizes TR binding to DNA by formation of TRAP-TR heterodimers with both proteins bound to the DNA. TR bound to the estrogen response element is unable to respond to TRAP and unable to stimulate transcription, possibly due to the absence of TRAP in the TR-estrogen response element complex. In addition, TRAP may interact with a certain subset of the nuclear receptor superfamily, since human retinoic acid receptor-beta and vitamin D receptor show increased binding to TREs in the presence of nuclear extract, but c-erbA alpha-2, a variant TR, does not respond to TRAP.

Animals↗

PubMed1990

A nuclear factor that enhances binding of thyroid hormone receptors to thyroid hormone response elements.

Recent studies from this laboratory have demonstrated the presence of thyroid hormone response elements (TREs) in the 5'-flanking region of the rat alpha and TSH beta subunit genes. Using an avidin-biotin complex DNA binding assay, we have shown that these TREs bind the thyroid hormone (T3) receptor present in nuclear extracts of GH3 cells, as well as the in vitro synthesized Hc-erbA beta, which has been identified as a member of the family of T3 receptors. The binding of Hc-erbA beta to the alpha subunit TRE can be enhanced 3-4-fold by including GH3 nuclear extract in the binding assay. Binding to the TRE present in the TSH beta gene or the rat growth hormone gene was similarly enhanced, although to a lesser degree. The enhanced binding activity is trypsin-sensitive and heat labile, and is not reproduced by the addition of histones, bovine serum albumin, or cytosol instead of nuclear extract. Gel exclusion chromatography suggests a molecular size of approximately 65,000 Da. This protein, which is present in several different cell types, is also able to complement binding of the rat erbA alpha-1 and the pituitary-specific erbA beta-2 forms of the receptor. These data suggest that the binding of the T3 receptor to a TRE is augmented by another nuclear protein, which may be involved in the mechanism of action of thyroid hormone.

Animals↗

PubMed1989

Thyroid hormone regulation of the rat glycoprotein hormone alpha-subunit gene promoter activity.

An important physiological control of the glycoprotein hormone alpha-subunit is the negative feedback by thyroid hormones in the thyrotrope. A region of the rat glycoprotein hormone alpha-subunit gene that is involved in transcriptional regulation by thyroid hormone has been identified by transient transfection studies, and sequence-specific binding of the thyroid hormone receptor to a site within this region has been demonstrated. Deletion-mutation studies using plasmid expression vectors containing either 246, 170, or 80 base pairs of the 5'-flanking region of the rat alpha-subunit gene fused to the coding region of the bacterial chloramphenicol acetyltransferase gene demonstrate 3,5,3'-triiodo-L-thyronine (T3)-regulated expression in GH3 cells, a T3-responsive somatotrophic cell line. In order to investigate the possibility of thyroid hormone receptor interaction with this segment of the rat alpha-subunit gene, the binding of the thyroid hormone receptor to synthetic oligodeoxynucleotides was analyzed using an avidin-biotin complex DNA binding assay. An oligodeoxyribonucleotide representing a fragment of the alpha-subunit gene from -74 to -38, relative to the transcriptional start site, shows significant binding to [125I]T3-receptor complex present in nuclear extracts of GH3 cells. This fragment binds receptor to a degree similar to that seen with a fragment of the rat growth hormone gene which contains a putative thyroid hormone-responsive element. In addition, this fragment of the rat alpha-subunit gene binds to the in vitro synthesized human c-erbA beta protein, which has been identified as a member of the family of putative T3 receptors. These data demonstrate that a cis-active thyroid hormone-responsive element resides in the 5'-flanking region of the rat alpha-subunit gene and that the mechanism involved in the suppression of expression of this gene by T3 could involve specific binding of the thyroid hormone receptor to this region of the gene.

Animals↗