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Biomedical subjects

S Spence

Publications and source records attributed to S Spence.

At least 37 records · Page 2Linked to original sources
PubMed1995

Effects of porcine growth hormone on pregnancy and fetal/neonatal development in the rat.

Porcine growth hormone was given subcutaneously twice daily to two groups of 20 females at dose levels of 0.5 and 2.5 IU/kg (1 and 5 IU/kg/day). A control group of 20 females was similarly treated with vehicle. The females were given either vehicle or porcine growth hormone from gestation day (GD) 6 through GD 21, for 10 females that were cesarean sectioned, or through lactation day (LD) 21, for 10 females scheduled for natural delivery. There were no deaths, abortions, or drug-related physical signs in any treatment group. Drug-related effects during gestation were limited to significant (p < or = 0.05) pharmacologically mediated increases in F0 maternal body weight gain during GD 6-20 in the 2.5-IU/kg group, approximately 24% above controls. During LD 0-21, there were significant (p < or = 0.05) dose-related increases in average maternal body weight gain in the 0.5- and 2.5IU/kg groups (72 and 200% above controls, respectively). Consistent with these findings, there were dose-dependent increases in maternal serum growth hormone and IGF-1 levels noted on GD 21 and LD 21 in both drug-treated groups. There were no drug-related effects on embryonal/fetal survival, GD 21 fetal body weight, placental weight, fetal femur length and width, or fetal morphology as determined by external, visceral, and skeletal examinations. There were no drug-related effects on F1 pup mortality, physical signs, body weight, biparietal diameter, liver weight, and femur length or width. These data suggest that subcutaneous administration of growth hormone to pregnant and lactating rats, at a dose that produces significant (p < or = 0.05) increases in maternal body weight gain and serum IGF-1 levels, has no apparent effect on embryonal/fetal development or preweaning growth.

Animals↗
PubMed1994

Ganciclovir treatment of herpes simplex thymidine kinase-transduced primary T lymphocytes: an approach for specific in vivo donor T-cell depletion after bone marrow transplantation?

Allogeneic bone marrow transplantation (BMT) is associated with a severe complication--graft-versus-host disease (GVHD). Although effectively preventing GVHD, ex vivo T-lymphocyte marrow depletion unfortunately increases graft rejection and reduces the graft-versus-leukemia (GVL) effect. The ex vivo transfer of the herpes simplex thymidine kinase (HS-tk) suicide gene into T cells before their infusion with hematopoietic stem cells could allow for selective in vivo depletion of these T cells with ganciclovir (GCV) if subsequent GVHD was to occur. Thus, one could preserve the beneficial effects of the T cells on engraftment and tumor control in patients not experiencing severe GVHD. To obtain T cells specifically depleted by GCV, we transduced primary T cells with a retroviral vector containing the HS-tk and neomycin resistance (NeoR) genes. Gene transfer was performed by coculturing PHA +/- CD3- or alloantigen-stimulated purified T cells on an irradiated retroviral vector producer cell line or by incubating the T cells in supernatant from the producer. Subsequent culture in G418 for 1 week allowed for the selection of transduced cells. GCV treatment of interleukin-2-responding transduced and selected cells resulted in greater than 80% growth inhibition, whereas GCV treatment of control cells had no effect. Similarly, the allogeneic reactivity of HS-tk-transduced cells was specifically inhibited by GCV. Combining transduced and nontransduced T cells did not show a bystander effect, thus implying that all of the cells inhibited by GCV were indeed transduced. Lastly, studies involving the transduction of the HUT-78 (T-lymphoma) cell line suggest that stable expression of HS-tk can be maintained over 3 months in vitro in the absence of G418. In summary, we have established the feasibility of generating HS-tk-transduced T cells for subsequent in vivo transfer with hematopoietic stem cells and, if GVHD occurs, specific in vivo GCV-induced T-cell depletion in allogeneic BMT recipients.

Bone Marrow↗
PubMed1994

Dietary linoleic acid-induced changes in respiratory beta-adrenergic receptor function and the form of arrhenius plots of isoprenaline- and prostaglandin E2-stimulated adenylate cyclase activity in a model for atopy.

Varying dietary linoleic acid altered lung membrane fatty acid composition with linoleic acid content increasing from approximately 6% total in those on 3 en% diet to approximately 14% total fatty acid in those on a 12 en% diet. Accompanying this were two- to three-fold increases in the levels of the elongation products of linoleic acid, namely 20:2 (n-6) and 22:5 (n-6) and a decrease in 18:1 oleic acid from approximately 26% to approximately 19% total. Administration of Haemophilus influenzae, to animals on 6 en% linoleic acid, serving as a model for atopy, effected a small increase in the levels of 22:5 (n-3) and doubled those of 22:6 (n-3). beta-Adrenergic-induced tracheal relaxation and stimulation of lung adenylate cyclase were elevated by increasing dietary linoleic acid from 3 to 6 en%, although such differences were abolished in the atopic model and when dietary linoleic acid was increased to 12 en%. Arrhenius plots of NaF-stimulated lung adenylate cyclase activities exhibited a break (t1) at approximately 26 degrees C in all dietary groups with unchanged activation energies and activity. In contrast, whilst both isoprenaline and PGE2-stimulated adenylate cyclase activities showed similar break-points in their Arrhenius plots, dietary linoleic acid manipulation markedly altered their form. As with NaF-stimulated activities then, irrespective of dietary manipulation and induction of atopy, these plots showed an invariant break occurring at approximately 26 degrees C. But, for animals on 3 and 6 en% diets, a second break was apparent at approximately 15 degrees C, which was slightly decreased to approximately 12 degrees C upon induction of atopy and completely abolished on increasing dietary linoleic acid to 12 en%. Accompanying such changes were marked alterations in activation energies. We suggest that profound changes in lung plasma membrane bilayer properties occur upon both altering dietary linoleic acid levels and in atopy. These selectively perturb adenylate cyclase activity when it is receptor-stimulated but not when it is activated by direct G-protein stimulation with NaF. We suggest that atopy and dietary challenge elicit an asymmetric perturbation of the plasma membrane that predominantly affects the outer half of the lipid bilayer.

Adenylyl Cyclases↗
PubMed1993

The role of Gi and the membrane-fluidizing agent benzyl alcohol in modulating the hysteretic activation of human platelet adenylate cyclase by guanylyl 5'-imidodiphosphate.

The non-hydrolysable GTP analogue guanylyl 5'-imidodiphosphate (p[NH]ppG) elicited a profound increase in the adenylate cyclase activity of human platelets. This occurred after a well-defined lag period of around 6 min, whereupon an enhanced steady-state rate was evident. The duration of the lag period was unchanged over a range of concentrations of p[NH]ppG which gave very different steady-state rates of adenylate cyclase activity. Prior activation of the stimulatory G-protein Gs by cholera-toxin pre-treatment abolished the lag period and elicited a small increase in the steady-state rate. Manipulating function of the inhibitory G-protein Gi also led to profound changes in the lag periods. Thus marked decreases in the lag were seen (approximately 70-81%) when Gi function was ablated through pre-treatment of platelet membranes with pertussis toxin, or by using elevated (25 mM) Mg2+ levels in the assay, or when Mg2+ was replaced by 5 mM Mn2+ in the assay. In contrast with this, potentiation of Gi function led to an increase in the lag period, as seen under conditions of agonist occupancy of inhibitory alpha 2-adrenoceptors (increase approximately 74%) or with the addition of 100 mM NaCl to the assays (increase approximately 44%). The local anaesthetic and membrane-fluidizing agent benzyl alcohol elicited both a profound decrease (around 70% at 80 mM) in the p[NH]ppG-induced lag period and a marked augmentation (around 5-fold) in the steady-state adenylate cyclase activity. When adenylate cyclase assays were done at 35 degrees C instead of 25 degrees C, then the lag period for activation by p[NH]ppG was decreased by around 33% and the steady-state rate increased by around 3-fold. At 35 degrees C, the addition of benzyl alcohol led to the apparent abolition of the lag period for p[NH]ppG activation of adenylate cyclase and amplified the steady-state rate by only around 2.2-fold. It is shown that Gi plays a fundamental role in determining the rate of activation of Gs. The proposal is formulated that such an action may be mediated through the release of beta gamma-subunits. Thus beta gamma-subunit dissociation is proposed as providing the rate-limiting step in Gi activation.

Adenosine Diphosphate Ribose↗
PubMed1990

Comparison of haematological values in restricted- and ad libitum-fed domestic fowls: white blood cells and thrombocytes.

1. White blood cells and thrombocyte values were examined in normal domestic fowls of layer and broiler strains fed restricted or ad libitum diets. 2. Basophils were the only white blood cells to increase significantly in number after the period of food restriction. It is suggested that the increase may have been associated with stress. 3. There were more basophils in broilers than layers, in males than females and in younger than older birds. 4. In contrast, a decrease in the total number of white blood cells was seen in birds maintained on a restricted diet compared with those fed ad libitum. 5. Stress may also have been responsible for an increased thrombocyte count in the restricted birds as well as a slightly lowered eosinophil count. 6. Heterophil/lymphocyte ratios were not significantly altered by the restriction of food.

Age Factors↗
PubMed1990

Comparison of haematological values in restricted- and ad libitum-fed domestic fowls: red blood cell characteristics.

1. A haematological study of the red blood cell characteristics in domestic fowls fed restricted and ad libitum diets was examined. The characteristics included Hb (haemoglobin), PCV (Packed Cell Volume), Rbc (Red blood cell count), MCH (Mean Cell Haemoglobin), MCV (Mean Cell Volume), MCHC (Mean Cell Haemoglobin Concentration) and reticulocytes. 2. The investigation showed that Hb, PCV, MCH and MCV were all reduced when broilers were fed a restricted diet. The number of Rbc's in these birds also increased significantly compared with control ad libitum-fed broilers. MCHC and reticulocyte numbers did not alter significantly. 3. It is proposed that some of the red cell characteristics may be restored to within "normal" ranges by food restriction. However, the results did show that there was a tendency to induce a microcytic normochromic anaemia in these restricted-fed birds.

Animal Feed↗
PubMed1989

The local anaesthetic benzyl alcohol attenuates the alpha 2-adrenoceptor-mediated inhibition of human platelet adenylate cyclase activity when stimulated by prostaglandin E1, but not that stimulated by forskolin.

Treatment of human platelets with concentrations of benzyl alcohol up to 50 mM augmented adenylate cyclase activity when it was assayed in the basal state and when stimulated by prostaglandin E1 (PGE1), isoprenaline or NaF. Benzyl alcohol antagonized the stimulatory effect exerted on the catalytic unit of adenylate cyclase by the diterpene forskolin. Benzyl alcohol did not modify the magnitude of the inhibitory response when the catalytic unit of adenylate cyclase was inhibited by using either low concentrations of guanosine 5'-[beta gamma-imido]triphosphate, which acts selectively on the inhibitory guanine nucleotide-regulatory protein Gi, or during alpha 2-adrenoceptor occupancy, by using adrenaline (+ propranolol). Some 34% of the potent inhibitory action of adrenaline on PGE1-stimulated adenylate cyclase was obliterated in a dose-dependent fashion (concn. giving 50% inhibition = 12.5 mM) by benzyl alcohol, with the residual inhibitory action being apparently resistant to the action of benzyl alcohol at concentrations up to 50 mM. Treatment of membranes with benzyl alcohol did not lead to the release of either the alpha-subunit of Gi or G-protein subunits. The alpha 2-adrenoceptor-mediated inhibition of adenylate cyclase was abolished when assays were performed in the presence of Mn2+ rather than Mg2+ and, under such conditions, dose-effect curves for the action of benzyl alcohol on PGE1-stimulated adenylate cyclase activity were similar whether or not adrenaline (+propranolol) was present. We suggest that (i) alpha 2-adrenoceptor- and Gi-mediated inhibition of PGE1-stimulated adenylate cyclase may have two components, one of which is sensitive to inhibition by benzyl alcohol, and (ii) the Gi-mediated inhibition of forskolin-stimulated adenylate cyclase exhibits predominantly the benzyl alcohol-insensitive component.

Adenylyl Cyclases↗
PubMed1989

Interaction of oxindanac and frusemide in man.

Oxindanac, a moderately active cyclo-oxygenase inhibitor in vitro, is a new antiinflammatory agent under clinical investigation. Its effects on frusemide-induced natriuresis have now been studied. Eight male volunteers receiving frusemide 40 mg b.d. were also given either oxindanac 300 mg b.d. or placebo in two consecutive periods separated by a treatment-free period, according to a randomized cross-over study design. Urinary prostaglandin excretion (PGF2 alpha) fell by 75% after 3 days on oxindanac. Frusemide-induced renin activity reached 66% of the control value in the presence of oxindanac. However, the natriuresis induced by frusemide did not differ significantly whether oxindanac or placebo was administered, despite the inhibitory action of the former on prostaglandin synthesis in vivo.

Adult↗
PubMed1989

Development of a realistic method to assess wheelchair propulsion by disabled people.

Most wheelchair users cannot achieve the steady state of cardio-respiratory performance necessary for standard physiological testing of wheelchair propulsion on ergometers or treadmills. Furthermore "real life" wheelchair utilisation involves short bursts of energy expenditure around furniture and other obstacles. We have developed a method of measuring wheelchair mobility on a test circuit of varying tortuosity and report here our preliminary experience. The parameters of mobility measured included distance; time taken; resting, maximum and final pulses (using a portable monitor); time to recover to stable resting pulse; and perceived exertion using the Borg Scale. Average speed and physiological cost of wheelchair propulsion (PCWP) (the difference between maximum and resting pulse divided by average speed) were calculated. The reproducibility of the components of the method was demonstrated in studies on normal volunteers. The test was also shown to be suitable for use by a sample of disabled people and was used in comparative studies of conventional and a novel arm crank wheelchair in normal volunteers. The method described appears to be an accurate and objective method of assessing wheelchair propulsion suitable for use by disabled people. It can also form the basis of comparative studies of different methods of propulsion of wheelchairs.

Adult↗
PubMed1981

Two cloned beta thalassemia genes are associated with amber mutations at codon 39.

Two beta globin genes from patients with the beta(+) thalassemia phenotype have been cloned and sequenced. A single nucleotide change from CAG to TAG (an amber mutation) at codon 39 is the only difference from normal in both genes analyzed. The results are consistent with the assumption that both patients are doubly heterozygous for beta(+) and beta degrees thalassemia, and that we have isolated and analyzed the beta degrees thalassemia gene.

Base Sequence↗
PubMed1981

Isolation and characterization of cloned DNA: the delta and beta globin genes in homozygous beta + thalassemia.

We have isolated and characterized a clone of human DNA from a patient with beta+ thalassemia containing the entire delta and beta structural genes and their flanking sequences. Partial Eco RI digestion of spleen DNA was used to obtain 15 to 20 kilobase (kb) pieces of human DNA that were then ligated to charon 4A lambda phage DNA. The 8 x 10(5) recombinants obtained were grown and screened for their content of beta gene sequences. Four positive clones were found, and one (beta T1-1) has been extensively analyzed. Subclones containing the entire beta gene and the large beta intervening sequence (IVS 2) have been isolated in the plasmid pBR 322. The fragments generated by restriction enzyme digestion in these subclones have been compared to those in similar subclones from normal beta genes. No differences have been found indication no significant rearrangements of deletions of the delta and beta genes. With the enzymes used, 11.2% of IVS 2 have been compared, and thus far no differences between the thalassemic and normal genes have been detected. The 24 enzymes used include Hph I, which recognizes the 5' end of IVS 2, and AIu I that cleaves at the 3' end. Thus, there appears to be conservation of nucleotide sequences at the ends of IVS 2 in this beta + thalassemia patient, although RNA metabolism studies suggest a possible defect in RNA processing.

Adult↗
PubMed1980

The organization of the gamma-delta-beta gene complex in normal and thalassemia cells.

Restriction enzyme digestion analysis and direct human globin gene cloning have permitted analysis of the physical arrangement of nucleotide sequences within and surrounding the human globin genes. With these methods it has been shown that the linear arrangement 5' to 3' of the globin genes is G gamma-A gamma-delta-beta. The G gamma and A gamma genes are separated by about 3.5 kilobases (kb), while the A gamma and delta genes are 15 kb apart, and the delta and beta 6.5 kb apart. Each of these genes contains a large intervening sequence (IVS) of approximately 1 kb in precisely the same position between condons 104 and 105. In addition, each of these genes has a small IVS between codons 30 and 31. In homozygous delta beta thalassemia DNA, there is deletion of all of the normal delta and beta gene fragments. However, a new fragment 4.2 kb in size containing the 5' end of the delta globin gene is retained. Retention of this fragment in delta beta thalassemia, but not in HPFH is consistent with a role for sequences in this region for limiting gamma globin gene expression. Studies to date suggest that the beta + and beta 0 thalassemias will be due to a heterogeneous group of DNA defects affecting either beta globin gene transcription or beta mRNA processing. In most cases of beta + and beta 0 thalassemia DNA analyzed, there is no detectable deletion of beta or delta genes. In three India beta 0 patients, deletion of the 3' end of the beta gene has been found. Analysis of cloned beta globin genes from a patient with beta + thalasseia shows differences from normal in the fragments generated by restriction enzymes which cut frequently. Whether these differences are responsible for the defect in thalassemia or are polymorphisms unrelated to thalassemia remains to be determined.

Base Sequence↗
PubMed1979

Isolation and characterization of cloned human fetal globin genes.

Three clones containing both the human G gamma and A gamma globlin genes have been isolated and characterized from a library of DNA fragments generated by partial Eco RI digestion of cellular DNA using charon 4A phage as vector. Two of the clones (NY 2 and 3) are identical and have an insert of 14.0 kb. The third clone (NY 1) has a 15.4 kb insert by virtue of an extra 1.4 kb Eco RI fragment at its 5' most end. This clone also has a Kpn I site not present in the other two suggesting it is the product of the gamma gene on the opposite chromosome. Restriction analysis of the three clones indicates that the G gamma and A gamma genes are linked on a single continuous piece of DNA and are separated by 3.5 kb and each contains at least one large intervening sequence of 0.85 kg between the Bam HI and Eco RI sites. These findings in cloned DNA provide direct evidence for linkage and organization of the gamma genes in man.

Base Sequence↗
PubMedDate not supplied

Teratogenic effects of the endothelin receptor antagonist L-753,037 in the rat.

The studies presented in this paper were undertaken to investigate the effects of L-753,037, a balanced endothelin receptor antagonist with similar affinity for the ET(A) and ET(B) receptors (Ki = 0.03 to 0.12 nM and 0.1 to 3.33 nM, respectively), when administered to pregnant female rats by oral gavage, on the development, growth, maturation, and reproductive performance of the F1 generation. Following embryonic exposure to L-753,037, profound craniofacial, cardiovascular, and viscerocranial malformations were noted in the F1 generation. All of the affected organs are derived, in part, from cranial neural crest cells predominantly originating from the posterior midbrain through the hindbrain and destined for the pharyngeal arches. In contrast, cranial structures derived from the paraxial mesoderm (i.e., basisphenoid) were of normal shape and size. There were no apparent effects on enteric neural crest cell derivatives. Based on the phenotype of affected fetuses and their similarity to fetuses with gene knockouts of ET-1 or the ET(A) receptor, the observed alterations are considered to be a pharmacologically mediated class effect on cranial neural crest cells. The phenotype observed suggests that ET(A) receptor antagonism may have specific effects on cranial neural crest cell migration and/or proliferation.

Abnormalities, Drug-Induced↗