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Biomedical subjects

J Porter

Publications and source records attributed to J Porter.

At least 163 records · Page 9Linked to original sources
PubMed1990

Dominant-negative mutants of a platelet-derived growth factor gene.

Using site-directed mutagenesis of a PDGF-A cDNA clone, we identify two domains that are required to generate stable, mitogenically active PDGF-AA homodimers. Alteration of the tetra-basic amino acid sequence (Arg84-Arg-Lys-Arg to Arg-Ser-Asn-Gly) results in the formation of stable pro-PDGF-A homodimers that lack mitogenic activity. Substitution of serine for Cys129 destabilizes PDGF-A subunits within the cell. Genes incorporating either the processing lesion or the cysteine substitution suppress wild-type PDGF-A gene expression in a trans-dominant fashion. Suppression occurs because the mutant PDGF subunits dimerize with wild-type subunits to form inactive or unstable heterodimers. Suppression is exerted across phylogenetic boundaries; thus, the mouse PDGF-A chain mutants inhibit the activity of the wild-type Xenopus PDGF-A. The cysteine mutant gene suppresses expression of PDGF-B (c-sis), as well as PDGF-A. The processing mutant gene, however, suppresses only PDGF-A. Dominant-negative mutations of PDGF and other growth factors which, like PDGF, function as dimers may prove useful for creating animals models of growth factor deficiency disease states and for revealing the function of growth factors during early embryonic development.

Amino Acid Sequence↗
PubMed1990

Randomized comparison of routine vs highly selective use of Doppler ultrasound and biophysical scoring to investigate high risk pregnancies.

OBJECTIVE: To compare routine versus highly selective use of Doppler ultrasound and biophysical scoring in higher risk pregnancy. DESIGN: A pragmatic randomized trial. SETTING: St James's University Hospital, Leeds. SUBJECTS: 500 pregnant women at high risk of intrauterine growth retardation or still birth. INTERVENTIONS: Regular monitoring with biophysical profile assessment and Doppler velocity waveform recording in umbilical and uteroplacental arteries. Results immediately available to clinicians. MAIN OUTCOME MEASURES: Gestational age at delivery, obstetric intervention rates and short-term neonatal morbidity. RESULTS: Risk factors were distributed very evenly between the 250 patients in the study and control groups respectively. A total of 902 biophysical profile and Doppler assessments were done in the 250 study group patients and only in 12 patients in the control group. In the study group, absent end-diastolic flow was found in only 2.7% of all 902 measurements. A persistently abnormal biophysical score was always associated with absence of end-diastolic flow. The mean gestational age at induction of labour was statistically and clinically similar in the two groups and there was no overall statistically significant difference in intervention rates between the two groups. There was a statistically significant lower frequency of depressed 5-min Apgar scores in the study group. Serious neonatal morbidity was also statistically significantly more common in the control group than in the study group. CONCLUSIONS: The use of Doppler ultrasound in higher risk pregnancies does not lead to an increase in iatrogenic preterm delivery. The total rate of positive tests on Doppler ultrasound is very low and persistently abnormal biophysical scores are unlikely to be found in patients where umbilical end-diastolic blood flow is present. Surrogate measures for fetal damage seem to be improved when clinicians have access to Doppler ultrasound assessments.

Adult↗
PubMed1990

Crucial role of pancreatic ductal collagenase injection for isolation of pancreatic islets.

We have been stressing the advantage of stationary digestion because of its simplicity and reproducible high yields of viable islets. In the present study optimal conditions of stationary in vitro collagenase digestion in mice and rats were examined. We also compared two possible routes for collagenase injection; ductal (PD) and portal venous (PV) based on subsequent islet yield and ability to reverse diabetes in rats. Three parameters which affect the quality of digestion are 1) collagenase concentration, 2) incubation time and 3) digestion temperature. Suitable conditions were easily determined and reproducible high yield of islets could be consistently obtained. The islets from one mouse pancreas (approximately 200 islets) could consistently restore normoglycemia in one STZ-induced diabetic mouse and the islets from one rat pancreas (500-600 islets) can restore normoglycemia in 5-6 STZ-induced diabetic mice within a couple of days. Islet yield in the PD method was greater than that in the PV method, and insulin release from PD islets in response to high glucose was well preserved after 24 hours of culture when compared to PV islets. The ability to restore normoglycemia in STZ-induced diabetic mice was well preserved when transplanting 100 PD islets as compared with the same number of PV islets. The PD islets revealed a well preserved structure with healthy endocrine cells, while the PV islets showed a dilated capillary network and distorted endocrine cell continuity. Histological examination of digested tissue following PD injection showed the complete destruction of pancreatic exocrine tissue, as well as mechanical separation and digestion of interstitial tissue between the islets and exocrine tissue, with the islet being preserved selectively intact.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

Gamma-irradiation as a tool to reduce immunogenicity of islet allo- and xenografts.

Sensitivity of pancreatic endocrine cells to gamma-irradiation and alteration of islet immunogenicity by irradiation were examined. Syngeneic, allogeneic (DBA/2) and xenogeneic (rat) islets were irradiated and transplanted at varying doses (8, 24, 40, 80 and 160 Gy) into streptozotocin-induced diabetic B6AF1 mice. In an isograft model, late loss of graft function was observed in some animals given 200 24-Gy-irradiated islets. However, larger numbers (400-500) of islets given 40 Gy irradiation did not show reversal of normoglycemia. With higher doses (80 or 160 Gy) significant early as well as late graft loss was observed. In an allograft model, the irradiated islet allografts survived beyond controls. Marked prolongation was achieved with a broad range of irradiation doses between 8 Gy and 120 Gy with 30-90% of recipients maintaining normoglycemia over 50 days. Late loss of graft function was observed between 78 and 180 days with a dose over 24 Gy. Increasing dose resulted in a better allograft survival rate in the early postoperative period, but tended to curtail longterm survival. In an xenograft model, irradiation of islets with 8 to 24 Gy led to prolongation of graft survival. Maximum graft prolongation was achieved with 24 Gy. Higher doses were much less effective and, in some recipients, caused shortening of graft survival.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Preparative-scale synthesis and reversed-phase purification of a gonadotropin-releasing hormone antagonist.

The preparation of "Nal-Glu" antagonist (Ac-D-Nal-D-Cpa-D-Pal-Ser-Arg- D-2-amino-5-oxo-5-(4-methoxyphenyl)pentanoic acid-Leu-Arg-Pro-D-Ala-NH2) was accomplished in two steps: (i) preparation of [Ac-D-Nal1, D-Cpa2, D-Pal3, Arg5, D-Glu6, D-Ala10]-GnRH via standard solid-phase synthetic techniques and (ii) acylation of anisole (Friedel-Crafts) by the glutamic acid residue in position 6. The preparative-scale, reversed-phase high-performance liquid chromatographic (HPLC) purification of the crude "Nal-Glu" antagonist employs first a triethylammonium phosphate (TEAP) (pH 2.25)-acetonitrile solvent system, followed by an HPLC-based desalting procedure, yielding the acetate salt of the peptide. This repetitive process of purification in TEAP-acetonitrile, followed by counter-ion exchange with 0.5% acetic acid-acetonitrile is highly reproducible and allows large amounts of a given peptide to be purified efficiently in a batchwise fashion. The procedure described for the synthesis, purification and characterization of the "Nal-Glu" antagonist is presented as a model for the multi-gram synthesis and purification of peptides to be used in clinical investigations.

Amino Acid Sequence↗
PubMed1989

Aerobic fitness in patients with fibrositis. A controlled study of respiratory gas exchange and 133xenon clearance from exercising muscle.

Aerobic fitness was evaluated in 25 women with fibrositis, by having them exercise to volitional exhaustion on an electronically braked cycle ergometer. Compared with published standards, greater than 80% of the fibrositis patients were not physically fit, as assessed by maximal oxygen uptake. Compared with matched sedentary controls, fibrositis patients accurately perceived their level of exertion in relation to oxygen consumption and attained a similar level of lactic acidosis, as assessed by their respiratory quotient and ventilatory threshold. Exercising muscle blood flow was estimated by 133xenon clearance in a subgroup of 16 fibrositis patients and compared with that in 16 matched sedentary controls; the fibrositis patients exhibited reduced 133xenon clearance. These results indicate a need to include aerobic fitness as a matched variable in future controlled studies of fibrositis and suggest that the "detraining phenomenon" may be of relevance to the etiopathogenesis of the disease.

Adult↗
PubMed1989

Successful treatment of experimental diabetes by sequential transplantations of multiple-donor pancreatic islet allografts.

Fifty hand-picked islets were freshly prepared from DBA/2 mice and transplanted four times into a streptozotocin-induced diabetic B6AF1 mouse without immunosuppression. Blood sugar levels decreased progressively and all recipients became normoglycemic after the 4th grafting. Four repeated transplantations at 2-4-day or 14-day intervals restored normoglycemia for more than 200 days in virtually all recipients. However, a majority of recipients given four transplantations of 50 DBA/2 islets at 7-day intervals or four transplantations of 100 DBA/2 islets at 4-day or 14-day intervals acutely rejected their grafts. When handpicked islets were freshly prepared from each of four histoincompatible donors (DBA/2, DBA/1, A.SW, and C3H) and sequentially transplanted four times (islets from one donor for each transplantation) into diabetic B6AF1 recipients, virtually all recipients maintained normoglycemia for more than 200 days regardless of the number of islets per grafting or intervals between transplantations. Since the purity of human islet preparations to date has been at the level of crude-digested islets, crude islets were used in sequential transplantation. Four transplantations of approximately 50 crude islets prepared from each of four donors achieved marked prolongation of graft survival. In sharp contrast, transplantation of 250-500 single-donor crude islets or four sequential transplantations of DBA/2 50 crude islets resulted in acute graft rejection.

Animals↗
PubMed1989

The gonadotropin-releasing hormone pituitary receptor interacts with a guanosine triphosphate-binding protein: differential effects of guanyl nucleotides on agonist and antagonist binding.

Binding of the GnRH agonist [DAla6,NMe-Leu7,Pro9Net]GnRH to bovine anterior pituitary membranes is inhibited by guanyl nucleotides. The effect of guanyl nucleotides is temperature dependent, in that significant binding inhibition is observed when the receptor-hormone interaction is measured at 37 C, and no inhibition is seen at 4 C. Micromolar concentrations of the nonhydrolyzable GTP analog 5'-guanylylimidodiphosphate [Gpp(NH)p] displace the bound agonist in a dose-dependent manner, with half-maximal displacement occurring in a concentration range of 0.1-0.5 microM, and maximum displacement occurring at a concentration of 50 microM Gpp(NH)p. At a concentration of 50 microM, the other nucleotides GTP and GDP inhibit binding to a lesser extent, while GMP, cGMP, 5'-adenylylimidodiphosphate [App(NH)p], ATP, and cAMP have no effect on the binding. At 37 C, Gpp(NH)p reduces the affinity of the agonist by a factor of 6 and increases its dissociation rate. In the presence of Gpp(NH)p at 37 C, there is also a 2-fold increase in the total number of binding sites. Under the same conditions as those used for the agonist, there is no displacement of the bound antagonist [Ac-D2Nal1,4ClDPhe2,D3Pal3,DLys6,Lys8,D Ala10]-GnRH by doses up to 50 microM Gpp(NH)p. The modulation of the binding of the agonist, but not that of the antagonist, by guanyl nucleotides is characteristic of receptors that are coupled to GTP-binding proteins. Thus, the GnRH receptor appears to be coupled to a GTP-binding protein that may play a role in the mechanism of action of GnRH at the pituitary.

Animals↗
PubMed1989

American perceptions of the British National Health Service: five myths.

This article explores five strong beliefs, or myths, held by Americans about the British National Health Service: (1) the NHS is socialized medicine; (2) widespread rationing occurs; (3) NHS patients have to face long waiting times; (4) the NHS does not offer free choice of provider; and (5) private medicine is taking over. The authors explore how ethnocentricity and American values have shaped these five myths, and argue that these cultural biases limit the ability of Americans to objectively evaluate the NHS and prevent them from learning from the British system.

Attitude to Health↗
PubMed1989

Effect of gamma-irradiation on mouse pancreatic islet-allograft survival.

Elimination or inactivation of lymphoid tissue in the pancreatic islet preparation achieves prolongation of islet-allograft survival. In this study we examined the effect of gamma-irradiation on mouse islet-allograft survival. In a B6AF1 isograft model, irradiation up to 2400 rad did not induce deterioration of islet function over 200 days, but greater doses caused cessation of graft function between 83 and 186 days. When DBA/2 crude islets were transplanted into B6AF1 recipients, all nonirradiated allografts were acutely rejected. Marked prolongation of allograft survival was achieved by islet irradiation with doses between 800 and 12,000 rad. With higher doses, significant numbers of allografts survived beyond the controls, but many lost function between 78 and 180 days, with none surviving greater than 200 days. Irradiation with 16,000 rad caused acute radiation damage. Because most secondary islet allografts in recipient mice that lost primary islet-graft function between 84 and 195 days survived greater than 100 days, late functional loss was probably due to the radiation injury. Combined use of recipient treatment with cyclosporin A and graft irradiation (2400 rad) achieved prolongation of DBA/2 islets in B6AF1 mice.

Animals↗
PubMed1989

Lower extremity peripheral neuropathy and ischemic ulcers associated with giant cell arteritis.

A patient with previously treated, temporal artery biopsy proven, giant cell arteritis (GCA) developed lower extremity ulcers and sensory neuropathy. Sural nerve biopsy showed epineural vessels with focal mild chronic inflammation. The lower extremity skin and nerve abnormalities improved after retreatment with prednisone. The rare manifestations of lower extremity skin and peripheral nervous system involvement associated with GCA are discussed. GCA must be considered in lower extremity ischemic processes of the skin and peripheral nervous system.

Foot Diseases↗
PubMed1989

Epidemiology of congenital syphilis.

Between 1984 and 1987, the number of reported cases of congenital syphilis in New Jersey tripled. Findings indicate an increase in early syphilis among females of childbearing age living in areas of high syphilis morbidity, reflecting, possibly, lifestyle changes within populations already at risk for the disease. Future studies and interventions are needed.

Adult↗
PubMed1988

Design of potent cyclic gonadotropin releasing hormone antagonists.

In order to improve the biological potency of cyclic gonadotropin releasing hormone (GnRH) antagonists, we have synthesized analogues, the conformations of which were restrained through internal side chain/side chain amide bridges linking aspartic acid or glutamic acid and L-2,3-diaminopropionic acid or L-ornithine. A disulfide bridge linking L-cysteine residues was also introduced. Residues belonging to the bridge spanned from position 4 to positions 9 or 10. Two series of analogues were synthesized and are characterized by residues at positions 1 [Ac-D-3-(2'-naphthyl)alanine], 2 [D-(4-chlorophenyl)alanine or D-(4-fluorophenyl)alanine], 3 [D-3-(3'-pyridyl)alanine or D-tryptophan], 5 (arginine or tyrosine), and 6 [D-3-(3'-pyridyl)alanine or D-arginine], respectively. These substitutions were selected in an effort to optimize high biopotency for inhibition of luteinizing hormone secretion, minimization of histamine release activity, and high (relative) hydrophilicity. The most potent analogues in the antiovulatory assay were cyclo(4-10) [Ac-DNal1,DCpa2,DPal3,(Asp4 or Glu4),Arg5,DPal]6,Dpr10]GnRH (compounds 5 and 7), which were fully active at ca. 12.5 micrograms/rat in the first series, and cyclo(4-10)[Ac-DNal1,DFpa2,DTrp3,Asp4,DArg6++ +,Dpr10]GnRH (compound 12), which was fully active at 2.5 micrograms/rat in the second.

Amino Acid Sequence↗
PubMed1988

Induction of antigen-specific unresponsiveness to pancreatic islet allografts by antilymphocyte serum.

The mechanism (or mechanisms) underlying the indefinite survival of DBA/2 islet allografts in strongly histoincompatible (C57BL/6xA)F1 (B6AF1) mice, induced either by the combined use of Ficoll-prepared crude islets and treatment of recipients with antilymphocyte serum (ALS), or by the use of handpicked, purified islets in nonimmunosuppressed recipients, was examined. B6AF1 mice bearing DBA/2 crude islet allografts for more than 100 days following ALS treatment accepted secondary DBA/2 crude islet allografts, but acutely rejected third-party A.SW crude islet allografts. This antigen-specific unresponsiveness to islet allografts can be successfully transferred into syngeneic B6AF1 mice by spleen cells. However, the state of unresponsiveness to donor antigen observed in these animals appears to be relatively weak, since transplantation of DBA/2 skin allografts or injection of DBA/2 spleen cells (5 x 10(7] caused acute rejection of long-term-accepted islet allografts. In contrast, B6AF1 mice bearing DBA/2 purified islet allografts over 100 days without immunosuppression rejected the secondary DBA/2 crude islet allografts acutely. Transfer of spleen cells obtained from these animals to syngeneic B6AF1 mice failed to induce prolongation of DBA/2 crude islet allografts. Thus, the mechanism (or mechanisms) involved in the long-term acceptance of islet allografts induced by the use of ALS and crude islets appears to be different from that involved in the long-term acceptance of purified islet allografts. The possible roles played by ALS in the induction of specific unresponsiveness to islet allografts are discussed.

Animals↗