Introduction and expression of human beta globin genes into primitive murine hematopoietic progenitor cells by retrovirus mediated gene transfer.
Explore the source record for details and available documents.
Animals↗
Biomedical subjects
S Karlsson
Publications and source records attributed to S Karlsson.
Failures and length of service in fixed prosthodontics after long-term function. A longitudinal clinical study.
In this long-term clinical study, the outcome of prosthetic restorative treatment was evaluated by examining fixed partial dentures 14 years after insertion. All prosthetic treatments had been performed by private general practitioners. The investigation comprised a total of 97 patients rehabilitated with 140 fixed reconstructions with and without cantilever extension. At the time of the examination 17% (24) of the original fixed restorations had been removed completely owing to loss of retention in most cases. Other technical failures were also recorded, including framework fractures and loose retainers. The total incidence of failure, biological ones excluded, was 26%. The incidence of failure for restorations with cantilever extension, was higher than that for fixed partial dentures with a distal abutment tooth. No significant difference in the rate of technical failure or loss of serviceability was found between the different age classes. After 14 years in service, 83% of the extensive fixed partial dentures were still functioning and fully serviceable, an overall result considered to be highly satisfactory.
Dental Abutments↗
Expression of human beta globin genes introduced into primitive murine hematopoietic progenitor cells by retrovirus mediated gene transfer.
Explore the source record for details and available documents.
Animals↗
Detection by high-performance liquid chromatography of polyamines formed by clostridial putrefaction of caseins.
Casein incorporated in building materials is degraded by species of alkali-tolerant Clostridia. A whole range of compounds have previously been detected in degraded building materials containing casein as an additive by gas chromatography (GC), including volatile and non-volatile organic acids, alcohols, ketones, aldehydes and monoamines. Using high-performance liquid chromatography (HPLC) it was possible, however, also to detect polyamines formed in degraded caseins. Histamine, agmatine, serotonine, tyramine, tryptamine, putrescine and cadaverine were detected in solutions containing casein in which the alkali-tolerant Clostriadia had been grown. Uninoculated, sterile incubated caseins contained no detectable amounts of polyamines. This gives clear evidence of the role of the biotic environment in the degradation of caseins. A combination of GC and HPLC therefore, provides a convenient set of techniques for studying the degradation products of casein.
Caseins↗
Expression of the human beta-globin gene following retroviral-mediated transfer into multipotential hematopoietic progenitors of mice.
Efficient transfer of the beta-globin gene into primitive hematopoietic progenitors was achieved with consistent and significant expression in the progeny of those cells. Retroviral vectors containing the intact genomic human beta-globin gene and the neomycin (G418)-resistance (neoR) gene were constructed. These gave titers of 10(6) or more neoR colony-forming units/ml when packaged in psi 2 cells. Mouse bone marrow cells were infected by coculture with producer cells and injected into lethally irradiated animals. Several parameters were varied to enhance infection frequency of colony-forming units, spleen (CFU-S); overall 41% of 116 foci studied contained an intact proviral genome. The human beta-globin gene was expressed in 31 of 35 CFU-S-derived spleen colonies that contained the intact vector genome at levels ranging from 1% to 5% of that of the mouse beta-globin genes. Infected bone marrow cells were also injected into genetically anemic W/Wv recipients without prior irradiation. Human beta-globin chains were detected in circulating erythrocytes by immunofluorescent staining with a specific monoclonal antibody. All animals injected with donor cells that had been cultured in G418 (1 mg/ml) for 48 hr after retroviral infection had circulating erythrocytes containing human beta-globin chains between 3 and 8 weeks after transplantation.
Animals↗
Follow-up study of patients with bridge constructions performed by private dental surgeons and at a university clinic, 8 years following insertion.
Patients treated in private practice and patients treated at the Department of Prosthetic Dentistry of the Faculty of Odontology in Gothenburg with fixed bridgework were questioned about their experience of their bridges. The interview was carried out with questionnaires and in some cases by telephone. The results were similar in both groups, and the patients were satisfied with the treatment.
Attitude↗
Effects of corticotropin-releasing hormone on insulin and glucagon secretion in mice.
Besides in the brain, corticotropin-releasing hormone occurs in the pancreas. Therefore, its effects on plasma levels of insulin and glucagon were investigated in vivo in the mouse. At 2 min after CRH injection (0.5-8.0 nmol/kg), plasma insulin was lowered: by 4.0 nmol/kg from 38 +/- 4 to 28 +/- 2 mU/l (P less than 0.05). Plasma insulin was lowered also at 6 min, whereas at 10 min, plasma insulin levels were elevated (P less than 0.05). Plasma glucagon levels were slightly lowered (P less than 0.05) at 10 min after CRH injection, whereas plasma glucose was slightly elevated (P less than 0.05) at 6 min after injection but not at 2 or 10 min. The effects of CRH on the plasma insulin and glucagon response to iv injections of half-maximal dose levels of glucose (2.8 mmol/kg) or the cholinergic agonist carbachol (0.16 mumol/kg) were also investigated. CRH, 4.0 nmol/kg, however, could not influence the plasma insulin or glucagon levels after the iv injection of either glucose or carbachol. Thus, CRH slightly affects basal plasma levels of insulin and glucagon in mice. In contrast, stimulated insulin and glucagon secretions are not affected by CRH. Peripheral CRH may therefore be of slight importance for the regulation of basal plasma levels of insulin and glucagon in the mouse.
Animals↗
Transfer of genes in hematopoietic cells with retroviral vectors.
Explore the source record for details and available documents.
Animals↗
A new moderately repetitive DNA sequence family of novel organization.
In cloning adenovirus homologous sequences, from a human cosmid library, we identified a moderately repetitive DNA sequence family consisting of tandem arrays of 2.5 kb members. A member was sequenced and several non-adjacent, 15-20 bp G-C rich segments with homology to the left side of adenovirus were discovered. The copy number of 400 members is highly conserved among humans. Southern blots of partial digests of human DNA have verified the tandem array of the sequence family. The chromosomal location was defined by somatic cell genetics and in situ hybridization. Tandem arrays are found only on chromosomes 4 (4q31) and 19 (q13.1-q13.5). Homologous repetitive sequences are found in DNA of other primates but not in cat or mouse. Thus we have identified a new family of moderately repetitive DNA sequences, unique because of its organization in clustered tandem arrays, its length, its chromosomal location, and its lack of homology to other moderately repetitive sequence families.
Adenoviruses, Human↗
Production of glycosylated physiologically "normal" human alpha 1-antitrypsin by mouse fibroblasts modified by insertion of a human alpha 1-antitrypsin cDNA using a retroviral vector.
Alpha 1-Antitrypsin (alpha 1AT) deficiency is a hereditary disorder characterized by reduced serum levels of alpha 1AT, resulting in destruction of the lower respiratory tract by neutrophil elastase. As an approach to augment alpha 1AT levels in this disorder with physiologically normal human alpha 1AT, we have integrated a full-length normal human alpha 1AT cDNA into the genome of mouse fibroblasts. To accomplish this, the retroviral vector N2 was modified by inserting the simian virus 40 early promoter followed by the alpha 1AT cDNA. Southern analysis demonstrated that the intact cDNA was present in the genome of selected clones of the transfected murine fibroblasts psi 2 and infected NIH 3T3. The clones produced three mRNA transcripts (5.8, 4.8, and 2.4 kilobases) containing human alpha 1AT sequences, secreted an alpha 1AT molecule recognized by an anti-human alpha 1AT antibody, with the same molecular mass (52 kDa) as normal human alpha 1AT and that complexed with and inhibited human neutrophil elastase. The psi 2 produced alpha 1AT was glycosylated, and when infused intravenously into mice, it had a serum half-life similar to normal alpha 1AT purified from human plasma and markedly longer than that of nonglycosylated human alpha 1AT cDNA-directed yeast-produced alpha 1AT. These studies demonstrate the feasibility of using a retroviral vector to insert the normal human alpha 1AT cDNA into non-alpha 1AT-producing cells, resulting in the synthesis and secretion of physiologically "normal" human alpha 1AT.
Animals↗
Retroviral-mediated transfer of genomic globin genes leads to regulated production of RNA and protein.
A high-titer amphotropic retroviral vector containing the neomycin resistance gene and a hybrid gamma-beta genomic human globin gene has been constructed. Mouse erythroleukemia cells infected with this virus were found to contain the full transcriptional unit of the transferred human globin gene by Southern blot analysis. These cells contain normally initiated, spliced, and terminated human globin mRNA. The human globin mRNA level increased 5- to 10-fold upon induction of the mouse erythroleukemia cells. Human globin chains were produced but only in a fraction of the cells as detected by immunofluorescent staining. A similar retrovirus containing a human beta-globin gene was used to transduce mouse erythroleukemia cells resulting in much higher levels of human globin synthesis than detected in mouse erythroleukemia cells transduced with the gamma-beta globin virus.
Animals↗
The mechanism of 2-deoxy-glucose-induced insulin secretion in the mouse.
1 Intravenous injection of 2-deoxy-glucose (3 mmol/kg) increases plasma levels of insulin and glucose in the mouse. This study investigated the contribution of various autonomic receptors to the plasma insulin response. 2 Pretreatment with the cholinergic antagonist methylatropine abolished the response completely. 3 Pretreatment with the alpha-adrenoceptor antagonist phentolamine caused a potentiated response, whereas the beta-adrenoceptor antagonist L-propranolol had no effect. 4 Pretreatment with methylatropine and phentolamine combined caused a small insulin response, which was inhibited by propranolol. 5 Pretreatment with the nicotonic antagonist hexamethonium abolished the response. 6 It is concluded that 2-deoxy-glucose induces a stimulation of insulin secretion in vivo in the mouse predominantly by a cholinergic mechanism. 7 2-Deoxy-glucose also activates alpha-adrenergic mechanisms which affect the size of the insulin secretory response. beta-Adrenoceptor and non-adrenergic, non-cholinergic mechanisms seem to be less important.
Animals↗
Recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) shortens the period of neutropenia after autologous bone marrow transplantation in a primate model.
The effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) on hematopoietic reconstitution after autologous bone marrow transplantation was evaluated in a primate model. Animals were given a continuous intravenous infusion of recombinant human GM-CSF for several days both before and after transplantation or only after the transplant procedure. Marrow ablation was accomplished by total body irradiation. In both groups of animals, the neutrophil count reached 1,000/mm3 by 8-9 d posttransplant compared with an interval of 17 and 24 d for two concurrent controls. After withdrawal of GM-CSF, neutrophil counts fell to values comparable to those observed in untreated controls. Accelerated recovery of platelet production was also observed in four of the five animals. Two additional animals were initially given GM-CSF several weeks posttransplantation because of inadequate engraftment. Prompt and sustained increases in neutrophil and platelet counts were observed. We conclude that GM-CSF may be useful in accelerating bone marrow reconstitution.
Agranulocytosis↗
Complications after percutaneous transluminal angioplasty in the iliac, femoral, and popliteal arteries.
During a 5-year period percutaneous transluminal angioplasty (PTA) was attempted in 134 iliac, femoral, and popliteal arteries in 127 patients. Complications were systematically registered and classified as major, minor, and radiologic. Seventeen major complications (12.7%), including three deaths, occurred, as well as five minor (3.7%) and 13 radiologic complications (9.7%). The last group showed no clinical symptoms. The frequency of complications in our series is high compared with that found in a survey of 13 studies with 2043 patients. The most marked discrepancy was our high frequency of septic symptoms. A mortality rate of 2.2% was also higher in our series, but we have consequently registered the 30-day mortality rate to make comparisons with surgical series relevant. Radiologic alterations are usually not reported in other series probably because they are regarded as a part of the method, but we consider these alterations as potentially dangerous and important to report because they can result in clinically relevant complications. Because of the complications PTA should only be performed in centers in which complications can be optimally treated.
Adult↗
Retroviral-mediated gene transfer into mammalian cells.
Retroviruses may be used as genetic vectors to transfer genes into mammalian cells with high efficiency. We have shown that the N2 vector will transfer a functional bacterial gene for neomycin resistance (NeoR) into more than 80% of mouse spleen foci. A derivative of the N2 vector was constructed to study transfer and expression of the human gene for adenosine deaminase (ADA) in mammalian lymphoid and hematopoietic stem cells. This vector, termed SAX, contains the human ADA cDNA with an SV40 promoter in addition to the NeoR gene. The SAX vector was found to efficiently transfer and express the ADA gene in an ADA-deficient human T-cell line. Gene transfer by SAX using an autologous nonhuman primate bone marrow transplant model resulted in expression of the human ADA gene in peripheral blood cells of treated animals. Human bone marrow treated with SAX produced 1%-2% of colonies in vitro that were expressing the vector genes. Transfer of genes into circulating hematopoietic stem cells of fetal sheep in utero was most efficient; vector gene expression was evident in 20%-40% of hematopoietic colonies. Therefore, retroviral vectors are capable of transferring functional genes into a wide variety of mammalian lymphoid and hematopoietic cells. Such vectors may be useful for clinical trials of gene therapy, that is, the correction of genetic diseases by insertion of a normal gene into a patient's defective cells.
Adenosine Deaminase↗
Regulated expression of human globin genes following transfer with retroviral vectors.
We have constructed retroviral vectors containing human globin genes (gamma-beta hybrid gene or beta globin gene) and have transferred these into mouse erythroleukemia cells and obtained regulated expression of human globin mRNA and protein. The level of human globin mRNA production is high (7-40% of endogenous mouse beta major mRNA) and human beta-globin chain production is also relatively high in these cells (see Karlsson et al, 1987, for details). Production of gamma-beta hybrid chains in the virally transduced cells is low despite high mRNA levels. This is due to instability of the gamma-beta chain and possibly also because the viral transcript can act as antisense globin RNA and reduce protein production. Some of our globin viruses have a high titer (up to 2 X 10(6) c.f.u./ml) and may be useful in transferring globin genes into bone marrow stem cells.
Animals↗
Effect of drug intake prior to or after meals on serum theophylline concentrations: single-dose studies with Euphylong.
Three separate single-dose, randomized, cross-over studies in normal, healthy volunteers were performed to investigate the possible influence of drug intake in relation to concomitant food intake and of different dietary habits such as cold and hot evening meals on the bioavailability of theophylline from Euphylong. Euphylong is a new, pH- and rpm-independent theophylline formulation designed for once-daily administration in the evening. The results obtained show that the timing of drug intake, in relation to concomitant food intake, has no influence on the bioavailability of Euphylong. Under equicaloric conditions, a 10% decrease in AUC and a 20% decrease in Cmax is observed when Euphylong is administered after a hot evening meal compared with a cold evening meal. Viewed against the dramatic changes in bioavailability of up to 100%, which have been observed for certain other sustained-release theophylline formulations, these minor changes for Euphylong are negligible and without clinical relevance.
Adult↗
Comparison of steady state serum theophylline concentrations in healthy volunteers after dosing with Euphyllin retard and PulmiDur.
In a randomized cross-over study, theophylline serum levels were compared over a 12-h dosing interval after repeated administration of Euphyllin retard and PulmiDur to 18 healthy male volunteers. Although both preparations are sustained-release formulations they differ in the systems used and in the state of hydration of theophylline. Two tablets each of Euphyllin retard or one tablet PulmiDur plus one tablet PulmiDur forte were administered in an identical dosing scheme always before food intake. The mean value curves of theophylline serum levels obtained at steady state clearly demonstrate the marked sustained-release properties of both preparations. The relative bioavailability of Euphyllin retard vs. PulmiDur calculated from the individual ratios of the dose adjusted AUCs was 96(83-111)%, results being given as geometric mean and 95% confidence limits. Hence Euphyllin retard is equivalent to PulmiDur with regard to the extent of absorption.
Adult↗