Biomedical subjects

J E Morgan

Publications and source records attributed to J E Morgan.

At least 145 records · Page 8Linked to original sources

PubMed1988

Phosphorylase kinase activities in damaged mouse skeletal muscles.

In the course of work in which the phosphorylase kinase (PhK)-deficient mouse was used as a model of a defined inherited myopathy, we measured the PhK activity in regenerated autografts of normal whole extensor digitorum longus (EDL) muscles. Initially, no PhK activity was found for up to 71 days after grafting. A more sensitive assay technique revealed PhK activity in regenerated normal grafts from 43 days after grafting, but the levels never reached those found in ungrafted normal muscle. PhK activity was also reduced in normal EDL muscles following either: denervation, or tenotomy, or denervation and devascularisation, or denervation, devascularisation and tenotomy, but the reduction was never as great as that observed in grafted muscle of equivalent age. PhK activity was also reduced in the tibialis anterior (TA) muscles of the myopathic C57B1/10 mdx strain of mouse, in which the skeletal muscles undergo persistent bouts of degeneration and regeneration, whilst retaining their vascular and nervous connections. It was concluded that the loss of PhK activity in grafted muscle is due to a combination of the effects of denervation, tenotomy and regeneration which occur on grafting.

Animals↗

PubMed1988

Myogenicity in vitro and in vivo of mouse muscle cells separated on discontinuous Percoll gradients.

Mouse muscle cells, obtained by enzymatically disaggregating newborn mouse muscle, were separated on a discontinuous Percoll gradient. The myogenicity in vitro of the resultant cell fractions was examined by counting the percentage of nuclei in myotubes. Myogenicity in vivo was assessed by implanting a cell suspension of one of the allotypes of glucose-6-phosphate isomerase (GPI) into a regenerating skeletal muscle graft of a second GPI allotype: the finding of hybrid GPI indicated that the implanted cells were myogenic. Separation of mouse muscle cells on a discontinuous Percoll gradient gave rise to two myogenic fractions, one of which was more myogenic in vitro than were the unseparated cells and one of which was less myogenic. Both of these fractions were myogenic in vivo. A cell fraction was also produced which was non-myogenic in vitro as well as in vivo. In vitro and in vivo measurements of myogenicity were therefore in broad agreement.

Animals↗

PubMed1988

Provocation-challenge studies in shrimp-sensitive individuals.

Thirty individuals with history of immediate, objective, adverse reactions after shrimp ingestion underwent double-blind, placebo-controlled shrimp-food challenges. All individuals who did not exhibit a positive response (reproduction of objective symptoms) were administered an open challenge of 16 whole cooked shrimp. Positive challenge responses occurred in 9/30 subjects (30%); six of these subjects experienced a positive response during the double-blind phase. Of the 21 remaining subjects, 12 experienced generalized pruritus as their only symptom, whereas nine subjects had completely negative challenge responses. All placebo challenges were negative. Although a positive skin test was strongly associated with challenge symptoms (p less than 0.001), the shrimp prick skin test titration end points were not different among the challenge groups. The serum shrimp RAST percent was significantly higher in the positive challenge group (p less than 0.02). Mean levels of shrimp-specific serum IgG, IgA, and IgM levels were not different among the challenge groups. Although no single immunologic variable could be consistently used to identify subjects more likely to exhibit a positive challenge response, the composite of a positive shrimp prick skin test and elevated serum shrimp-specific IgE (RAST percent label bound greater than 11%) demonstrated a correct predictive value of 87% in this group of shrimp-sensitive subjects.

Allergens↗

PubMed1988

Earthworms as biological monitors of cadmium, copper, lead and zinc in metalliferous soils.

Earthworms (Lumbricus rebellus and Dendrodrilus rubidus) were sampled from one uncontaminated and fifteen metal-contaminated sites. Significant positive correlations were found between the earthworm and 'total' (conc. nitric acid-extractable) soil Cd, Cu, Pb and Zn concentrations (data log1) transformed). The relationships were linear, and the accumulation patterns for both species were similar when a single metal was considered, even though there were species difference in mean metal concentrations. Generally, the earthworm Cd concentration exceeded that of the soil; by contrast, the worm Pb concentration was lower than the soil Pb concentration in all but one (acidic, low soil Ca) site. Our observations suggest that Cu and Zn accumulation may be physiologically regulated by both species. Total-soil Cd explained 82-86% of the variability (V2) in earthworm Cd concentration; 52-58% of worm Pb and worm Zn concentrations were explained by the total-soil concentrations of the respective metals. Total-soil Cu explained only 11-32% of the worm Cu concentration. The effect of soil pH, total Ca concentration, cation-exchange capacity (CEC) and organic carbon on metal accumulation by L. rubellus and D. rubidus was investigated by multiple regression analysis. Soil pH (coupled with CEC) and soil Ca had a major influence on Pb accumulation (V2 of worm Pb increased to 77-83%), and there was some evidence that Cd accumulation may be suppressed in extremely organic soils. The edaphic factors investigated had no effect on Cu or Zn accumulation by earthworms. In the context of biomonitoring, it is proposed that earthworms have a potential in a dual role: (1) as 'quantitative' monitors of total-soil metal concentrations (as shown for Cd); and (2) as estimators of 'ecologically significant' soil metal, integrating the effects of edaphic factors (as shown for Pb).

Journal Article↗

PubMed1988

Calcium-lead interactions involving earthworms. Part 2: the effect of accumulated lead on endogenous calcium in Lumbricus rubellus.

The calcium and lead burdens of tissue fractions of Lumbricus rubellus were quantified in 'native' animals from acidic and calcareous disused lead mines, and from control ('naive') animals exposed to lead-polluted soils under laboratory conditions. Most of the body burden of lead was accumulated within the posterior alimentary canal, and significant positive correlations were generally found between the calcium and lead burdens in this tissue fraction, which were evident in both the naturally and laboratory lead-exposed animals. The calcium:lead correlation is probably due to a proliferation of the calcium-rich, lead-sequestering chloragosome granules, and may thus be regarded as a specific tissue response to cellular lead incursion. No calcium-lead relationship was recorded in the rest (largely composed of the body wall) fraction of earthworms inhabiting the lead-polluted sites. However, a concomitant increase in calcium and lead in this tissue fraction of the laboratory lead-exposed control animals was noted. It is concluded that in naturally lead-exposed earthworms, the cells of this tissue fraction may be relatively resistant to the toxic effects of the metal. By contrast, it is apparent that a non-specific cytotoxic response by the cells of the rest of 'naive' animals occurs, as demonstrated by the concurrent increase in its calcium and lead burdens. These results suggest that a tolerance mechanism to lead, perhaps with a genetic basis, may exist in earthworms naturally exposed to lead.

Journal Article↗

PubMed1988

Calcium-lead interactions involving earthworms. Part 1: The effect of exogenous calcium on lead accumulation by earthworms under field and laboratory conditions.

Earthworms (Lumbricus rubellus and Dendrodrilus rubidus) were collected from several acidic and calcareous abandoned ferrous metalliferous mine sites. Tissue lead concentrations were substantially lower than the total soil lead concentrations, except at one site (Cwmystwyth) where the tissue lead concentrations of both species were approximately 5 to 10 times higher than that of the soil. Soil lead was the major factor in determining the tissue lead concentration, although it was demonstrated that both soil pH and soil calcium concentration could markedly increase the % variance in tissue lead concentration. These findings help explain the apparent anomaly in tissue lead concentrations of earthworms from Cwmystwyth, where the soil is acidic and has exceptionally low calcium concentrations. Soil-liming experiments provided supportive evidence that soil pH, coupled with soil calcium, influences lead accumulation by earthworms, but a filter paper feeding experiment provided unequivocal evidence that soil calcium concentration alone can influence lead accumulation by earthworms. It is concluded that, although lead accumulation by earthworms is influenced by both physico-chemical and biochemical mechanisms, the latter over-rides the former, i.e. soil calcium is more important factor in determining the accumulation of lead earthworms than is soil pH.

Journal Article↗

PubMed1988

Molecular and genetic mapping of the mouse mdx locus.

mdx is an X-linked muscular dystrophy mutant of the mouse and a putative homolog of the human X-linked muscular dystrophy locus--Duchenne muscular dystrophy (DMD). Utilizing a C57BL/10/Mus Spretus interspecific cross in which the mdx mutation was segregating, we have constructed a detailed genetic map around the mdx locus on the mouse X chromosome. We were unable to detect recombinants between mdx and exonic probes derived from the human DMD gene. These genetic data support the contention from biochemical studies (E.P. Hoffman, R. H. Brown, and L. M. Kunkel, 1987, Cell 51: 919-928) that DMD and mdx are homologous genes.

Animals↗

PubMed1987

Polyamines and acetylpolyamines increase the stability and alter the conformation of nucleosome core particles.

The interactions of spermine (4+ charge at physiological pH), N1-acetylspermine(3+), spermidine(3+), N1- and N8-acetylsperimidine(2+), putrescine(2+), hexaamminecobalt(3+), and magnesium(2+) with nucleosome core particles have been examined by using thermal denaturation and circular dichroism. Tetra- and triamines were 2-3 times more effective than diamines at stabilizing core particles against thermal denaturation. Secondary effects were also observed, with acetylpolyamines slightly less effective than unmodified polyamines of equivalent charge. Hexaamminecobalt(3+) was less effective than the triamines, while magnesium had essentially no effect. This is surprising since magnesium is more effective than diamines at stabilizing naked DNA. All the cations tested altered the circular dichroism spectra of the core particles in the DNA region (284 nm). The peak at 284 nm was suppressed by tetra- and trivalent compounds to approximately twice the extent of divalent compounds. Magnesium appears to suppress the peak by a lesser extent than the diamines. This indicates that the DNA twist and/or folding is changed by these cations. A plateau of both thermal denaturation and circular dichroism effects was observed at cation concentrations where 30-40% of the total DNA negative charges could be neutralized by the added cations. We suggest that polyamine and histone acetylation function in concert to lower the stability and change the conformation of the nucleosome core, thus facilitating replication and transcription in vivo.

Animals↗

PubMed1987

Spatial properties of neurones in the lateral geniculate nucleus of the pigmented ferret.

We used quantitative electrophysiological techniques to study the spatial properties of single units recorded extracellularly in the lateral geniculate and perigeniculate nuclei of the adult pigmented ferret. All neurones examined had approximately circular receptive fields, whose central regions gave responses antagonistic to those elicited from the surrounds. We presented sinusoidally modulated grating patterns, either drifting or counterphased, to obtain spatial frequency tuning curves, contrast sensitivity functions and to assess linearity or non-linearity of each neurone's response. In the ferret, as in other species, two types of lateral geniculate neurone could be distinguished, and we have termed these X-cells and Y-cells; both groups responded briskly to visual stimulation but X-cells gave sustained and linear responses whereas Y-cells responded transiently and non-linearly. Perigeniculate cells gave linear responses. For neurones in the lateral geniculate and perigeniculate nuclei, both the limit of spatial resolution ('acuity') and optimum spatial frequency were inversely related to receptive field eccentricity and the diameter of the receptive field centre. We recorded geniculate neurones in the ferret with acuities up to 3 cycles deg-1 and contrast sensitivities up to 114, values that are lower than those found previously for many geniculate cells in the cat.

Animals↗

PubMed1987

The movement of muscle precursor cells between adjacent regenerating muscles in the mouse.

Regeneration of mature skeletal muscle fibres involves the formation of new multinucleate muscle fibres by the fusion together of mononucleate muscle precursor cells. Such precursor cells appear to be largely or entirely derived from satellite cells, located between the basement membrane and the sarcolemma of the muscle fibre. We have previously presented evidence that precursor cells which contribute to regenerating muscle in a region of muscle damage are not all locally derived but that some migrate in from exogenous sources. The present study examines the possibility that a regenerating muscle might receive muscle precursor cells from neighbouring muscles. To do this we have made whole muscle allografts in the mouse and used the two murine isoenzyme allotypes of the dimeric enzyme Glucose-6-Phosphate Isomerase (GPI) as markers to demonstrate whether there is movement of muscle precursor cells between these allografts and adjacent host muscles. In host muscles adjacent to some allografts, a "hybrid" form of GPI was detected, each molecule consisting of one donor and one host GPI subunit. Such heterodimers can form only where host and donor nuclei share a common cytoplasm: in muscles this means that mosaic host/donor muscle fibres are present. The presence of such fibres implies that muscle precursor cells must have migrated into the host muscle from the neighbouring allograft.

Animals↗

PubMed1987

Thermal denaturation of mononucleosomes in the presence of spermine, spermidine, N1-acetylspermidine, N8-acetylspermidine or putrescine: implications for chromosome structure.

Putrescine (a diamine) raises the thermal denaturation temperature of mononucleosomes but produces only minor changes in the overall shape of the thermal denaturation curve. This is similar to the effect of sodium ions and is consistent with nonspecific binding to the DNA of the nucleosome. At very low levels of spermidine or spermine the same simple rise in thermal denaturation temperature is seen but at higher levels (above 1 microM for total spermidine concentration) the thermal denaturation curve becomes substantially sharper and the premelt region of the curve diminishes in area. The acetylspermidines display intermediate effects. The change in shape of the thermal denaturation curve was resolved into components (R1 and R2) due to mononucleosomes in their original conformation plus a component (T) induced by the presence of spermidine or spermine. The proportion of component T was substantially reduced with acetylspermidine, compared to equivalent concentrations of spermidine. Hence, we suggest that spermidine acetylation in vivo has the potential to partially destabilise the nucleosome structure, possibly in coordination with histone acetylation.

Acetylation↗

PubMed1987

Muscle precursor cells invade and repopulate freeze-killed muscles.

A problem with the use of muscle grafting as a therapeutic procedure is to produce a graft functionally adequate to replace a muscle of complex architecture, such as a sphincter muscle. We thought it might be possible to use dead cadaver muscles, repopulated by the patient's own muscle precursor cells (mpc), to reconstruct muscles whose anatomy would be imposed by the framework of dead muscle and whose genetic constitution would be determined by the mpc. Here we show, in the mouse, that an extensor digitorum longus (EDL) muscle, killed by repeated freezing and thawing, repopulated with mpc and grafted into a nu/nu or tolerant AKR host mouse, is capable of supporting muscle formation. By using the allotypic isoenzyme forms of glucose-6-phosphate isomerase as markers, we have shown that the newly regenerated muscle in such grafts is derived mainly from the implanted mpc, but also to some extent from the host mouse's own mpc. By 50-70 days after grafting, new muscle fibres were found to constitute up to 70% of the graft. Many fibres had assumed diameters in the normal range for mouse muscle, often having peripherally placed nuclei. These findings raise the possibility of the therapeutic use of such grafts. To our surprise, dead EDL muscle grafts into which no mpc had been implanted were also the site of good muscle regeneration. New-formed muscle in these grafts was shown to be derived entirely from mpc which must have migrated into the graft from the host. Investigation of the mechanisms underlying this phenomenon should further our knowledge of factors which regulate the proliferation and movement of dormant mpc in adult animals.

Animals↗

PubMed1987

Immunologic evaluation of shrimp-allergic individuals.

Thirty-three individuals with a history of immediate hypersensitivity reactions after shrimp ingestion and 29 nonshrimp-sensitive control subjects were evaluated for evidence of crustacea-specific immunity by skin prick test titration end point, RAST, and ELISA, with extracts of shrimp, crab, crayfish, and lobster. Individuals were categorized as either atopic or nonatopic on the basis of history and skin test reactivity to common inhalant allergens. Most (28/33) shrimp-sensitive subjects had positive skin prick tests to shrimp extract, whereas skin tests were negative in 27/29 control subjects. Eighty-one percent of atopic and 41% of nonatopic shrimp-sensitive subjects had elevated shrimp-RAST ratios. The RAST ratios of atopic individuals were significantly higher than ratios of nonatopic individuals, and there was a significant correlation between shrimp-RAST ratios and historical clinical symptom scores. RAST determinations of all control subjects were negative. Shrimp-sensitive subjects also had significantly elevated serum levels of shrimp-specific IgG and IgA as compared to control individuals. Both IgG and IgA shrimp-specific reactivity demonstrated a significant positive correlation with shrimp-RAST ratios. These studies indicate that IgE-mediated, type I mechanisms, detected by positive shrimp skin tests and RASTs, appear to be operative in crustacea-sensitive individuals, particularly those with concurrent respiratory allergy. Although the role of shrimp-specific IgG and IgA antibodies in the immunopathogenesis of crustacea allergy remains unclear, such antibodies appear to represent increased immunologic recognition of shrimp allergens/antigens in shrimp-sensitive subjects.

Adult↗

PubMed1987

Retinal decussation patterns in pigmented and albino ferrets.

The decussation patterns of retinal ganglion cells in adult pigmented and albino ferrets were determined from the distribution of cells labelled after large unilateral injections of horseradish peroxidase into the visual pathway, involving the lateral geniculate nucleus and fibres of passage to the superior colliculus. About 6000 retinal ganglion cells project ipsilaterally in pigmented ferrets compared with only about 1500 in albino ferrets. In both strains, the vast majority of these cells (99 and 87% in pigmented and albino animals, respectively) are located in the temporal crescent, although we describe one albino ferret in which an aberrant uncrossed projection arises from nasal retina. In pigmented ferrets, there is a sharp nasotemporal division that runs through the area centralis; a small proportion of the ganglion cells in temporal crescent (less than 10%) does project contralaterally. In albinos, however, the majority of cells in temporal retina project contralaterally. There is no clear nasotemporal division in the albino retina; the density of uncrossed ganglion cells is reduced throughout temporal crescent and at no location exceeds the comparable density of the crossed projection. The peak density within the reduced uncrossed projection is also displaced away from the area centralis into temporal retina. Analysis of cell type on the basis of soma size indicates that whereas large horseradish peroxidase injections into the visual pathway of pigmented ferrets label all types of ganglion cell in the crossed projection, injections restricted to the superior colliculus label only those ganglion cells with large or small somata. The distribution of cell sizes in the crossed projection from temporal retina is biased towards small cells in the pigmented ferret but in albinos resembles that seen in the crossed projection from nasal retina. Thus the adult pigmented ferret has both a well developed nasotemporal division in which decussation lines are obvious in the crossed and uncrossed pathways and also, unlike rodents but like cats, a class of ganglion cell that does not project to the superior colliculus. The albino mutation both reduces the uncrossed projection throughout temporal retina, although the reduction is greatest close to the area centralis, and also commensurately increases the crossed projection from temporal retina.

Albinism↗

PubMed1987

Use of the cutaneous anaphylactoid test to detect differences in mast cell mediator-releasing activity among LHRH peptides.

The cutaneous anaphylactoid test was evaluated as a screen for assessing mast cell mediator-releasing (MCMR) activity of luteinizing hormone-releasing hormone (LHRH) peptides. As expected, LHRH and D-Trp6-LHRH, an agonist of LHRH, were nonreactive at concentrations of antagonist which induced pronounced skin lesions. Differences between LHRH and antagonists were obvious, but it was difficult to detect differences among antagonists using arithmetic means. However, when the geometric means of the highest positive dilution were calculated, the MCMR activities of the antagonists differed and these differences appeared to be related to the hydrophobicities of their N-termini. Thus, it appears that the cutaneous anaphylactoid test is an appropriate screen to assess the MCMR activity of LHRH peptides and should assist in the development of clinically useful inhibitory analogs of LHRH.

Anaphylaxis↗

PubMed1987

Pulmonary immunologic features of alveolar septal amyloidosis associated with multiple myeloma.

A 74-year-old man presented with interstitial pulmonary disease which was proven to be alveolar septal amyloidosis by transbronchial biopsy. Multiple myeloma was diagnosed on the basis of monoclonal IgG-lambda protein in serum, monoclonal lambda light chains in urine, a bone marrow plasmacytosis of 22 percent, and serum IgA and IgM levels less than 100 mg/dl and 50 mg/dl, respectively. Appropriate investigations failed to show additional sites of deposition of amyloid. Analysis of fluid from bronchoalveolar lavage showed an increase in total cells recovered, a lymphocytosis with a ratio of T helper over T suppressor cells greater than that in peripheral blood, the presence of an IgG-lambda paraprotein, and an IgG/albumin ratio greater than that in serum. While plasma cells could not be identified in the recovered cell population, cultured cells from bronchoalveolar lavage fluid showed increased production of IgG. These findings provide evidence of an ongoing pulmonary immune response resulting in excess IgG-lambda protein in the pulmonary compartment, a factor which may contribute to the development of amyloidosis.

Aged↗

PubMed1986

Relationship between the displacement of phosphatidate phosphohydrolase from the membrane-associated compartment by chlorpromazine and the inhibition of the synthesis of triacylglycerol and phosphatidylcholine in rat hepatocytes.

Glycerolipid synthesis was studied in isolated hepatocytes by using 177 microM [14C]oleate and 1 mM [3H]glycerol. Chlorpromazine (25-400 microM) inhibited the synthesis of phosphatidylcholine and triacylglycerol. This was accompanied by an average increase of 12-fold in the accumulation of the labelled precursors in phosphatidate at 200 microM chlorpromazine and a decrease in the conversion of phosphatidate to diacylglycerol of 76%. These results indicate that part of the inhibition of the synthesis of phosphatidylcholine and triacylglycerol occurs at the level of phosphatidate phosphohydrolase. The relative rate of triacylglycerol synthesis at different concentrations of chlorpromazine was approximately proportional to the rate of conversion of phosphatidate to diacylglycerol. Phosphatidylcholine synthesis increased at higher rates of conversion of phosphatidate to diacylglycerol, but it was relatively independent of the latter rate when this was inhibited by more than about 30% with chlorpromazine. The addition of oleate to the hepatocytes caused a translocation of phosphatidate phosphohydrolase from the cytosol to the membrane-associated compartment. Chlorpromazine had the opposite effect and displaced the phosphohydrolase from the membranes in the presence or absence of oleate. There was a highly significant correlation between the activity of phosphatidate phosphohydrolase that was associated with the membranes of the hepatocytes and the calculated conversion of [3H]phosphatidate to diacylglycerol. Chlorpromazine also antagonized the association of the phosphohydrolase with microsomal membranes when cell-free preparations were incubated with combinations of oleate and spermine. Furthermore, it inhibited the transfer of the soluble phosphohydrolase to microsomal membranes that were labelled with [14C]phosphatidate and thereby decreased diacylglycerol production. It is concluded that part of the action of chlorpromazine in inhibiting the synthesis of triacylglycerol and phosphatidylcholine occurs because it prevents the interaction of the soluble phosphatidate phosphohydrolase with the membranes on which glycerolipid synthesis occurs. This in turn prevents the conversion of phosphatidate to diacylglycerol.

Animals↗

PubMed1986

CB 1954 revisited. II. Toxicity and antitumour activity.

We have assessed the antitumour activity of the nitrophenylaziridine CB 1954 in vitro and in vivo. For EMT6 mouse mammary tumour multicellular spheroids under hypoxic conditions in vitro, a 6-h exposure to 40 micrograms/ml reduced the surviving fraction to as low as 10(-3) and the growth delay was 5.4 days. Oxic cells were twofold less sensitive. Phenyl AIC protected oxic and hypoxic cells equally. Under oxic conditions minimal cell killing was seen with HT29 cells, either in multicellular spheroids or in monolayer; a 6-h exposure to 40 micrograms/ml gave a spheroid growth delay of 1.5-1.7 days. No growth delay was seen with single maximum tolerated doses of CB 1954 against HT29 grown as a xenograft in immunosuppressed mice. Only minimal growth delays of 1-2 days were seen with similar doses against the EMT6 tumour and the RIF-1 and KHT sarcomas in mice. Little activity was seen with maximum tolerated doses given once a day for 5 days against EMT6 and RIF-1. No chemosensitization was measurable with CCNU, cyclophosphamide or melphalan in the KHT tumour.

Animals↗