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Biomedical subjects

A B Murray

Publications and source records attributed to A B Murray.

At least 19 recordsLinked to original sources

Retrovirus-like particles from the human T47D cell line are related to mouse mammary tumour virus and are of human endogenous origin.

Retrovirus-like particles were secreted in a steroid-dependent manner by the human mammary carcinoma cell line T47D. The particles exhibited typical retroviral properties such as their electron microscopic appearance (95 nm in diameter) and occasional budding, sedimentation at 1.14 g/ml, reverse transcriptase activity and genomic RNA. The T47D particles were related to mouse mammary tumour virus (MMTV) as shown by their ultrastructural appearance (B type-like eccentric dense cores and budding), Mg2+ dependence of the reverse transcriptase activity; immunological reactivity with MMTV-directed antibodies (revealing proteins of 63K, 52K, 26K and 18K), and hybridization of particle RNA with MMTV DNA under stringent conditions. Purified particles were able to incorporate deoxynucleoside triphosphates in the absence of an exogenous primer and template, thus indicating the existence of a complete and biochemically functional reverse transcription apparatus (reverse transcriptase, RNA and primer) and the ability to direct endogenous cDNA synthesis. Labelled particle cDNA hybridized strongly to human genomic DNA but not to mouse and cat DNA, thus indicating the human origin of the T47D particles. Furthermore all human DNAs, hybridized with the labelled particle cDNA, showed a uniform hybridization pattern of restriction fragments, indicating the endogenous origin and distribution of the proviral particle DNA in the human genome.

Adenocarcinoma

Sexual dimorphism of the kidney in the NMRI mouse as shown by Dolichos biflorus agglutinin labelling.

The histological affinity pattern of Dolichos biflorus agglutinin (DBA) in kidneys from mice (NMRI, Balb/c, CBA) and rats (Wistar) fixed by perfusion with formalin, Bouin, or HgCl2 was investigated with a horseradish peroxidase conjugate. The animals were examined from fetal stage to adulthood. Adult female NMRI mice exhibited constant DBA labelling, with DBA binding to cells of the proximal and collecting tubules. Moreover the vascular endothelium of the renal papilla was found to be DBA-positive in 50% of adult female animals. In contrast, there was only very little DBA binding in the kidneys of male adult NMRI mice. There was no sexual dimorphism in lectin labelling in kidneys from other strains of mice or from rats.

Animals

Effect of passive smoking on asthmatic children who have and who have not had atopic dermatitis.

We studied 240 children with asthma who were themselves nonsmokers and had been referred consecutively to our clinic. They were aged 6 to 17 years. The severity of asthma was assessed by symptom score, by spirometry, and, in those who could perform the test reliably, by histamine bronchial challenge test. Those who reported having had a chronic or chronically relapsing itchy rash in characteristic locations were recorded as having had atopic dermatitis. Multiple analysis of variance revealed that children whose mothers smoked had significantly more severe asthma (p less than 0.001) but that atopic dermatitis had no apparent effect on the severity of asthma, either in its main effect (p = 0.71) or in its interaction with maternal smoking (p = 0.66). Although our previous study indicates that smoking mothers' children are more likely to develop asthma if they have had atopic dermatitis than if they have not, the severity of asthma does not appear to be associated with a history of atopic dermatitis. In smoking mothers' children the asthma was just as severe in those who had not had atopic dermatitis as in those who had.

Adolescent

Intradermal vaccination against hepatitis B in a group of medical students.

A prospective study of a low-dose (one-tenth) intradermal regimen using recombinant hepatitis B vaccine was undertaken during two consecutive years in 4th-year medical students. Eighty-one per cent of the vaccinees (123/152) sero-converted with anti-HBs levels of greater than 10 IU/l. The lower titre of hepatitis B surface antibodies compared with published studies on intramuscular immunisation, together with a sero-conversion rate of only 81%, makes the intradermal method, in our opinion, a suboptimal form of hepatitis B immunisation.

Hepatitis B Antibodies

Bone tumor induction after incorporation of short-lived radionuclides.

Temporally-limited internal irradiation after incorporation of short-lived bone-seeking radionuclides is a useful experimental tool for the investigation of extrinsic and intrinsic factors which modify the dose dependence of bone tumor risk. Here we describe some of the results obtained in experiments with female mice (mainly NMRI). The future aim of such experiments should be the prediction of risk of late effects using early molecular-biological changes. Molecular-biological descriptions in our model are at present very limited.

Animals

In situ embedding of cell monolayers cultured on plastic surfaces for electron microscopy.

Various procedures suitable for routine in situ embedding of cell monolayers were tested including: (1) the use of different Epon substitutes, (2) the use of different types of plasticware obtained from different sources, and (3) different methods of preparing capsules for sectioning. Different resins reacted differently with different plastics and type of preparation. Merck Epon substitute bound to most of the plastics tested. Ladd Epon substitute released cleanly from all plastics tested when a suitable method of preparation was used. The results show that for routine embedding of cell monolayers it is necessary to select an appropriate Epon substitute and method of preparation of capsules for the type of plasticware used. A routine method is described, with various alternative steps which can be applied when particular difficulties are encountered.

Animals

Effects of Nd: YAG laser coagulation of myocardium on coronary vessels.

Laser coagulation of arrhythmogenic myocardium proved to be an effective surgical therapy for refractory arrhythmias. To determine the influence of Nd: YAG laser coagulation of myocardium on coronary vessels, a total of 48 transcatheter laser impacts (10 W, 10 sec, 7 W mm2) were directed to the left ventricular free wall via endocardial (24 lesions in 2 dogs) or epicardial (24 lesions in 2 dogs) approach. In 8 of 24 endomyocardial lesions (2 and 6 months old) coronary vessels with a diameter of greater than 50 microns were found within the coagulation zones. The volume of these lesions was significantly (P less than 0.01) smaller (139 +/- 43 mm3) than those (n = 16) with coronary vessels measuring less than 50 microns (311 +/- 87 mm3). Volumes of epimyocardial lesions (1-2 hours old) produced by transcoronary irradiation with normal coronary blood flow were significantly (P less than 0.01) smaller (31 +/- 17 mm3) than with reduced (73 +/- 22) or interrupted (119 +/- 34 mm3) blood flow (n = 8, each). Both directly irradiated coronary vessels and those found within the coagulation zones and scars appeared histologically normal through all layers with an intact intima without fibrosis or thrombosis. The ultrastructure of irradiated arteries, even with total interruption of blood flow, was no different from that of non-irradiated controls. Thus, coronary artery blood flow significantly reduces the volume of coagulated myocardium, whereas the vessels themselves appear to remain undamaged by laser irradiation as used for myocardial coagulation.

Animals

Bone formation by osteoblast-like cells in a three-dimensional cell culture.

Cells of the clonal osteogenic cell line MC3T3-E1 were seeded onto a three-dimensional matrix of denatured collagen type 1 and cultured for a period of up to 8 weeks. Specimens were analyzed by histological, enzyme histochemical, immunocytochemical, and ultrastructural methods and by in situ hybridization between day 7 and day 56 after seeding. In 56-day cultures, the MC3T3-E1 cells were arranged in a three-dimensional network and formation of bone-like tissue was indicated by calcification of a newly synthesized collagen type I matrix resembling osteoid and surrounding osteocyte-like cells. The differentiating culture showed high expression of osteocalcin and alkaline phosphatase activity. NIH3T3 fibroblasts used as control cells passed through the network of the substrate forming a confluent monolayer underneath. This culture system offers a potentially powerful model for bone formation in vitro and for investigating the osteogenic potential of bone-derived cells.

Alkaline Phosphatase

It is children with atopic dermatitis who develop asthma more frequently if the mother smokes.

We elicited symptoms of atopic dermatitis (AD) and of asthma from 620 children who were themselves nonsmokers, were aged 1 to 17 years, and had been consecutively referred to an allergy clinic. A histamine bronchial-challenge test revealed bronchial hyperresponsiveness in 95% of the children who had a history of wheezing or asthma and who could perform the test reliably, indicating that most of them did indeed have asthma. Children with a history of AD were much more likely to have asthma if the mother was a smoker than if she was a nonsmoker (79% versus 52%; p = 0.001). Similarly, if AD was found on examination, the percentages with asthma were 74% and 44%, respectively. By contrast, the children with no history of AD had asthma as frequently if the mother was a nonsmoker (42%) as when she was a smoker (40%). In children with AD, the prevalence of asthma was greater in both boys and girls when the mother was a smoker, but only in boys when the father was a smoker. Multiple logistic regression confirmed that the risk of asthma was greatly increased when the child had both AD and a mother who smoked.

Adolescent

c-fos expression precedes osteogenic differentiation of cartilage cells in vitro.

We have investigated the temporal pattern of expression of c-fos in cartilage cells in mouse mandibular condyles. During in vitro cultivation, the progenitor cells in this organ differentiate to osteoblasts, and hypertrophic chondrocytes start to show features indicative of osteogenic differentiation. Prior to these processes we observed two distinct patterns of c-fos expression. High, transient c-fos expression was found in the entire tissue within 30 min of culture. This type of c-fos expression appeared to result from mechanical forces applied during dissection. The second type of c-fos expression appeared in individual cells in the zone of hypertrophic chondrocytes. A varying number of formerly quiescent chondrocytes expressed high levels of c-fos mRNA after between 30 min and 10 d in culture, with a peak in the number of cells between days 1 and 3. c-fos expression in these cartilage cells was followed by DNA replication and expression of genes typifying osteoblastic differentiation. After 7 d in culture, groups of cells with the typical ultrastructural features of osteoblasts, and surrounded by an osteoid-like matrix, were observed in single chondrocyte-type lacunae, suggesting division of chondrocytes and differentiation to osteoblasts. The data suggest that c-fos may play a crucial role in the perturbation of determined pathways of skeletoblast differentiation and in the regulation of endochondral bone formation.

Animals

Induction of lymphoma and osteosarcoma in mice by single and protracted low alpha doses.

Internal doses from the short-lived alpha-emitter 22Ra were given to 4-wk-old female NMRI mice. One group of about 300 animals received a single injection of 18.5 kBq 22Ra kg-1 body weight, corresponding to a mean skeletal alpha dose of 0.15 Gy. A second group of about 300 animals received the same total amount of 224Ra in the form of 72 fractions of 257 Bq kg-1 each, applied twice weekly during 36 wk. The fractionated group received the same final mean total skeletal dose of 0.15 Gy as the single injected group, but with a mean skeletal dose rate of 1 mGy d-1. A rather high incidence, 13.5% (40/296), of early malignant lymphomas was observed in the fractionated group during and shortly after the injection period, followed by a 7% incidence (21/296) of osteosarcomas during the second half of the animals' lifetime. The group with a single injection did not develop early lymphomas but did develop osteosarcomas later with an incidence of 5.8% (17/295). The occurrence of osteosarcomas was similar up to day 800 in the two experimental groups. Surprisingly, however, after this period no additional case of osteosarcoma was observed in the single-injected group, whereas one-third of all osteosarcomas occurred after day 800 in the protracted group. The additional later occurrence of osteosarcomas occurred after indicates a longer lasting induction effect on osteosarcomas, or a promoting effect in older age, for this kind of treatment.

Alpha Particles

Acidophilic macrophage pneumonia in laboratory mice.

Acidophilic macrophage pneumonia, characterized by an accumulation of characteristic crystalloid-laden alveolar macrophages, was seen in 30/7,500 NMRI, 7/600 T x HT, 2/100 C57BL and in no cases of 1,500 CBA and 1,100 BALB/c mice. Histologically, there was a focal accumulation of large numbers of eosinophilic macrophages, generally associated with granulocytes. Macrophages could be mononucleate or multinucleate and had a crystalline cytoplasm. Free-lying crystals were sometimes observed. Ultrastructurally, macrophages had a cytoplasmic accumulation of needle-shaped and rhomboidal crystals, often showing a clear lattice structure with a repeat of 3-5 nm. The crystalloid inclusions may be derived from the breakdown products of granulocytes and appear similar to inclusions in macrophages in other parts of the hematopoietic system. That these inclusions are probably derived from eosinophils is based on the appearance within macrophages of structures resembling eosinophil granules at various stages of degradation and the similarity between the lattice repeat of the crystalloids and that of the crystalline core of the eosinophil granule. The crystalloid inclusions may be related to the Charcot-Leyden crystals found in human beings.

Age Factors

Osteosarcoma risk after simultaneous incorporation of the long-lived radionuclide 227Ac and the short-lived radionuclide 227Th.

The effect of injection of 1.85 kBq/kg of the long-lived radionuclide 227Ac on the induction of osteosarcomas in female NMRI mice by different dose levels (18.5, 74, and 185 kBq/kg) of the short-lived radionuclide 227Th was investigated. The highest absolute osteosarcoma incidence was observed with the highest doses of 227Th. Addition of 227Ac resulted in an additional osteosarcoma incidence only at the lowest dose of 227Th and did not affect the osteosarcoma incidence resulting from higher doses of 227Th. The longest times to tumor appearance were observed with 227Ac alone. The latent period in two different age groups (4 weeks and 10-12 weeks) appeared to be similar following injection with combined doses of 227Th and 227Ac but different after injection of each radionuclide alone.

Actinium

Passive smoking by asthmatics: its greater effect on boys than on girls and on older than on younger children.

In 415 nonsmoking asthmatic children who were seen consecutively, asthma symptoms were more severe if the mother was a smoker than if she was a nonsmoker. This applied to both sexes but was more marked in boys than in girls. There were also other indications that sons were the more severely affected: the forced expiratory volume at 1 second, the forced expiratory flow rate during the middle half of the forced vital capacity, and the provocation concentration of histamine needed to result in a 20% decrease in the forced expiratory volume at 1 second were significantly decreased only in the sons, and lung function test results were significantly less in sons than in daughters of mothers who smoked. When the 415 children were stratified according to age, lung function improved significantly with increasing age in the children of nonsmokers; in children of smokers, by contrast, symptoms and lung function test results became progressively worse. As well, there was a correlation between these indications of asthma severity and the number of years the child had been exposed to the mother's smoke. It appeared that, compared with girls, boys were more sensitive to passive smoking, and that its adverse effect increased with age and with duration of exposure.

Adolescent

Sudden outbreak of a leukemia-like lesion in female CBA mice after repeated injections of 5-azacytidine.

A total of 11 i.p. injections of 1 mg/kg 5-azacytidine given within 21 weeks induced a sudden outbreak of a leukemia-like disease in female CBA mice about 2 weeks after the last injection. The outbreak of disease was highly synchronous; 68% of animals who had survived the treatment period succumbed within a period of 5 days, 2/3 of them on a single day. The lesion had a characteristic form commonly presenting as a result of hemorrhagic effusion in the thoracic and/or abdominal cavities, associated with diffuse infiltration of lymphatic and fatty tissue by lymphoblasts. It appears that a critical combination of factors was responsible for the unexpected appearance of the lesion. Female BALB/c mice treated identically were not affected. The factors indicating that the lesion was induced by an epigenetic mechanism are discussed.

Animals

Establishment and characterization of osteogenic cell lines from a spontaneous murine osteosarcoma.

Five clonal cell lines were established from a spontaneous BALB/c mouse osteosarcoma, and characterized. Four of these lines showed some similarities in morphology, in vitro growth properties, production of collagenous and noncollagenous extracellular matrix proteins and osteogenic differentiation. The cells formed colonies with characteristic differences in size and morphology in soft agar, and osteogenic sarcomas and metastases in syngeneic mice after transplantation. Ultrastructurally, cells in the transplant tumours showed marked osteogenic features. There were no osteoclast-like cells. The fifth cell line had somewhat different characteristics. All five lines expressed infectious endogenous murine leukemia viruses. Increased c-myc protoon-cogene expression was found in one cell line and c-fos expression at different levels in all lines. There was only very low expression of c-Ha-ras and no expression of c-Ki-ras and c-sis. DNA analysis showed the presence of newly acquired proviral genomes integrated at different sites in the cellular DNA. The results show that distinct osteogenic neoplastic subclones can be obtained from a primary mouse osteosarcoma. Although the clones exhibited an appreciable morphological, functional, and molecular diversity they retained the basic pathogenic properties of the tumour from which they were derived.

Alkaline Phosphatase