Biomedical subjects

R D Jolly

Publications and source records attributed to R D Jolly.

At least 73 records · Page 4Linked to original sources

PubMed1989

Lipopigments in veterinary pathology: pathogenesis and terminology.

The lipopigments are a heterogenous group of pigments whose pathogenesis and terminology is confused. Whereas there is epidemiological and observational evidence that ceroid is derived from degeneration and peroxidation of unsaturated lipid, the assumption that all so-called lipopigments are similarly formed, is questioned. In particular, recent studies have distanced the pathogenesis of the pigment found in the ceroid-lipofuscinoses from that perceived for ceroid. The importance of protein rather than lipid in the pathogenesis of the pigment of ceroid-lipofuscinosis and of age pigment from the equine thyroid is noted. In the former the essential feature is storage of the DCCD binding protein subunit c of mitochondrial ATP synthase. There is a need for more analytical studies on isolated pigments which are generally more soluble than anticipated by the literature. It is proposed that the term ceroid be limited to a family of pathological pigments where lipid degeneration and peroxidation is implied from observational and/or epidemiological factors. The term age pigment is unequivocal and preferred for age related pigment not obviously complicated by other factors. The terms lipofuscin and lipopigment retain a usefulness as generic terms, particularly where the nature of the pigment is uncertain. The term ceroid-lipofuscinosis for the inherited storage diseases of children and animals is misleading. The term "proteolipid proteinosis" has been suggested to define this group of diseases but this is perhaps premature until their full pathogenesis is known.

Aging↗

PubMed1989

Lysosomal storage of the DCCD reactive proteolipid subunit of mitochondrial ATP synthase in human and ovine ceroid lipofuscinoses.

The ceroid lipofuscinoses (Batten's disease) are a group of neuro-degenerative lysosomal storage diseases of children and animals that are recessively inherited. In the diseased individuals fluorescent storage bodies accumulate in a wide variety of cells, including neurons. The material stored in the cells of sheep affected with ceroid lipofuscinosis is two-thirds protein. The stored material does not arise from lipid peroxidation or a defect in lipid metabolism, and the lipid content is consistent with a lysosomal origin for the storage bodies. The major protein stains poorly with Coomassie blue dye and is soluble in organic solvents. It has an apparent molecular weight of 3,500 and its amino acids sequence is identical to that of the dicyclohexylcarbodiimide (DCCD) reactive proteolipid, subunit c, of mammalian mitochondrial ATP synthases. Apart from removal of mitochondrial import sequences, it has not been modified post-translationally. At least 50% of the mass of the storage bodies is composed of this protein. A minor protein sequence related to the 17-kDa subunit of vacuolar H(+)-ATPase is also found in storage bodies isolated from pancreas. As in humans and cattle, the ovine protein is the product of two expressed genes named P1 and P2. In normal and diseased animals there are no differences in sequences between P1 cDNAs or P2 cDNAs, nor do levels of mRNAs in liver for P1 or P2 differ substantially between normal and diseased animals. Both normal and diseased sheep also express a spliced pseudogene encoding amino acids 1 to 31 of the mitochondrial import presequence. The peptides they encode differ by one amino acid; arginine-23 is changed to glutamine in the diseased sheep. Storage bodies isolated from brains and pancreas of children affected with the juvenile and late infantile forms of ceroid lipofuscinosis also contain large amounts of material that is identical to subunit c of ATP synthase. However, the protein is not present in storage bodies isolated from brains of patients affected with the infantile form of the disease, and these storage bodies contain other unidentified proteins. It is possible that the cause of ovine, juvenile and late infantile ceroid lipofuscinoses is related to a defect in degradation of the subunit c of mitochondrial ATP synthase.

Amino Acid Sequence↗

PubMed1989

Glycoconjugates in storage cytosomes from ceroid-lipofuscinosis (Batten's disease) and in lipofuscin from old-age brain.

The ceroid-lipofuscinoses (CL) are a group of inherited diseases characterised by the accumulation, in brain, of autofluorescent storage cytosomes which have similar histochemical staining properties to lipofuscin, the neuronal wear and tear pigment of old-age brain. The storage cytosomes stain strongly with periodic acid-Schiff reagent (PAS), indicating the presence of carbohydrate. In brain from each childhood form of CL, concentrations of phosphorylated dolichol (Dol-P) are 10- to 20- fold higher than in age-matched controls. Brain Dol-P concentrations are also increased between 2 and 5- fold in several different lipidoses and in elderly subjects. Much of the Dol-P which accumulates is located within the storage cytosomes. Dol-P constitutes 2-3% of the dry weight of storage cytosomes from juvenile and late-infantile CL, and 0.3-0.7% of storage cytosomes from infantile CL, ovine CL and of lipofuscin isolated from old age brain. The bulk of the Dol-P in CL brain and in isolated storage cytosomes is present as dolichyl pyrophosphoryl oligosaccharides (Dol-PP-OS). The constitutions of the oligosaccharide moieties differ in the various forms of the disease. Histochemical analysis of frozen sections of unfixed brain after extraction by various lipid solvents indicates that the major part of the PAS positive intraneuronal material in CL brain and in old-age brain has the extraction properties of Dol-PP-OS. Carbohydrate represents 4-7% of the dry weight of CL storage cytosomes and of lipofuscin. The major monosaccharide components are mannose, N-acetyl glucosamine, glucose and galactose. Depending on the form of the disease studied, up to 40% of this material can be accounted for by Dol-PP-OS. Polyacrylamide gel electrophoresis of storage cytosomes followed by lectin blotting demonstrates several low molecular weight components which bind concanavalin A. These do not coelute with the major protein components and may well be Dol-PP-OS. We conclude that Dol-PP-OS are concentrated in storage cytosomes in CL and are one of their major glycoconjugate components.

Aging↗

PubMed1989

A method of cerebro-cortical biopsy in lambs.

A suction needle brain biopsy technique is a safe and effective method of obtaining cerebral cortex for experimental purposes in lambs. The biopsies are performed under general anaesthesia using halothane and oxygen following barbiturate induction. The method may have a number of related used in this of other species.

Journal Article↗

PubMed1988

Ovine ceroid-lipofuscinosis. I: Lipopigment composition is indicative of a lysosomal proteinosis.

The ceroid-lipofuscinoses are inherited lysosomal storage diseases of children and animals characterised by a fluorescent lipopigment stored in a variety of tissues. Defects in lipid metabolism or the control of lipid peroxidation have been postulated to explain their pathogenesis but the underlying biochemical defect is still unknown. In the present study lipopigment was isolated from liver, kidney, pancreas and brain of sheep affected with ceroid-lipofuscinosis. Approximately two-thirds of the lipopigment mass was protein. Sodium dodecyl sulphate polyacrylamide gel electrophoresis showed a major polypeptide band of Mr 14,800, heterogeneous polypeptides between 5,000-9,000 Mr and a major band of Mr 3,500. These were not normal lysosomal proteins. I125 radiolabeling studies indicated that they were 47% of the pancreatic lipopigment mass, the 3,500 Mr polypeptides alone accounting for 26%. Lipopigment polypeptides were not subunits of a larger protein held together by disulphide bonds. The presence of the 3,500 Mr proteins in whole affected tissue homogenates distinguished them from homogenates of normal tissues. Lipopigment phospholipids were the same species as normal lysosomal phospholipids, including bis (monoacylglycero) phosphate, a lysosomal marker. Similarly the neutral lipids, notably dolichol, ubiquinone and dolichyl esters were typical of those in lysosomal membranes. Lipopigments contained 1-1.7% metals. Analyses of them indicated a functional lysosomal origin for the lipopigment. It was concluded that low Mr proteins are specifically stored in ovine ceroid-lipofuscinosis and that this disease is a lysosomal proteinosis.

Animals↗

PubMed1988

Ovine ceroid-lipofuscinosis II: Pathologic changes interpreted in light of biochemical observations.

Ceroid-lipofuscinosis is studied in a breed of sheep and the histochemical and ultrastructural changes are interpreted in light of the known chemical composition of cytosomes. It is concluded that the ovine disease is a lysosomal proteinosis. It is postulated that the multilamellar arrays reflect a complex three-dimensional structure of alternating lipid and protein with the proteinaceous component being specific polypeptides. This is supported by powder X-ray diffractions at 10A implying a repeating proteinaceous unit in the structures. The coherent size of diffracting units was remarkably similar to the periodicity of lamellae observed in electron micrographs of lipopigment cytosomes. The fluorescent and staining characteristics of lipopigment can be explained by the above conclusion and postulate.

Animals↗

PubMed1987

Mannosidosis: ocular lesions in the bovine model.

The ocular pathology of mannosidosis was studied in the bovine model. There was wide-spread vacuolation of many cell types including corneal epithelium, Descemet's endothelium, corneal fibroblasts, pigmented cells, lens epithelium, lens fibres, pigment epithelium and all cell types of the neuroretina. On electron-microscopy most vacuoles were seen to be membrane bound vesicles compatible with being secondary lysosomes and similar to those previously described elsewhere in the body. Additional vacuoles were seen due to dilatations between lens epithelial cells and between lens fibres. The cause of lens and corneal opacities seen in human patients is unclear from the present study but are presumably a consequence of the lesions noted.

Animals↗

PubMed1986

Screening for carriers of genetic diseases by biochemical means.

For some genetic diseases the underlying biochemical anomaly is known. Through the gene dosage phenomenon it may therefore be possible to detect the more numerous clinically normal heterozygotes and so initiate a control programme. Such programmes need to be carefully and individually planned according to certain general principles derived in part from experience with prototype programmes. Laboratory data may be interpreted in relation to the prior probability of an individual being heterozygous or normal which may be known from the status of parents or other close relatives. Instigation of a control programme based on heterozygote testing is best achieved by working through a breed society (or its equivalent) which can control pedigree breeding through the control of registrations.

Animal Husbandry↗

PubMed1986

The objective interpretation of histopathological data: an application to the ageing of ovine bruises.

Muscle and adipose tissue from a total of 178 experimental bruises inflicted on sheep and aged from 1 to 72 h old were processed for light microscopic examination. Five observed histopathological features of inflammation and repair were scored semiquantitatively on a scale of 1-4 according to their degrees of change from the normal state. These data were evaluated mathematically using a Bayesian probability model designed for the purpose. The model was able to age bruises with an acceptable degree of accuracy only as either 1-20 h or 24-72 h old but within these constraints a degree of confidence in excess of 90% was achieved. The exact performance of the model depended on the nature and number of tissue samples examined but mathematical ageing was superior to interpretative ageing based on personal experience.

Adipose Tissue↗

PubMed1986

Ceroid lipofuscinosis in sheep. I. Bis(monoacylglycero)phosphate, dolichol, ubiquinone, phospholipids, fatty acids, and fluorescence in liver lipopigment lipids.

The ceroid lipofuscinoses are inherited lysosomal diseases of children characterized by a fluorescent lipopigment stored in a variety of tissues. Defects in lipid metabolism or the control of lipid peroxidation have been postulated to explain their pathogenesis. In the present study, lipopigment was isolated from the liver of sheep affected with ceroid lipofuscinosis. It was 70% protein, the rest being mainly lipids. These were only one-sixth as fluorescent as total liver lipids, but contained a number of fluorophors. None were major components of the lipopigment or the postulated fluorescent product of lipid peroxidation. Lipopigment lipids included the lysosomal marker bis(monoacylglycero)phosphate that contained 42.9% linoleate and 16.5% linolenate. Lipopigment neutral lipids were dolichol, dolichyl esters, ubiquinone, free fatty acids, and cholesterol, indicative of a lysosomal origin of the lipopigment. Phosphatidylcholine, phosphatidylinositol, phosphatidylserine, and phosphatidylethanolamine were present in proportions and with fatty acid profiles typical of lysosomes. No differences were found between the lipids of total control and affected livers, nor the fatty acid profiles of their phosphatidylcholine, phosphatidylethanolamine, or triglycerides. It is concluded that ovine ceroid lipofuscinosis is not a lipidosis, nor does the lipopigment arise from the abnormal peroxidation of lipids. Strong similarities between the lipopigment and the age pigment lipofuscin were noted.

Animals↗

PubMed1986

Ceroid lipofuscinosis in sheep. II. The major component of the lipopigment in liver, kidney, pancreas, and brain is low molecular weight protein.

Previous studies on the lipopigment from the livers of sheep affected with ceroid lipofuscinosis showed that the disease does not involve a defect in lipid metabolism or abnormal lipid peroxidation and that most of the lipopigment was proteinaceous. In this study, lipopigment was isolated from liver, kidney, pancreas, and brain of affected sheep without the use of proteolytic enzymes. Lipopigment from all tissues was two-thirds protein. Modified silver staining after sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed a major band of Mr = 14,800, heterogeneous material between Mr = 5,000 and 9,000, and a major band of Mr = 3,500. These compounds did not stain for RNA or carbohydrate and were digested by a nuclease-free protease as expected for protein. They are not normal lysosomal proteins. Lipopigment levels of dolichol, ubiquinone, and cholesterol were consistent with the lipopigment being protein-enriched lysosome-derived cytosomes. The presence of the Mr = 3,500 proteins in whole affected tissue homogenates distinguished them from homogenates of normal tissues. It was concluded that low Mr proteins are specifically stored in ovine ceroid lipofuscinosis and that the ceroid lipofuscinoses may result from inherited defects in lysosomal protein catabolism.

Animals↗

PubMed1986

A defect in thyroglobulin synthesis in an inherited ovine goitre: possible neonatal respiratory distress syndrome.

A ram lamb with congenital goitre showed signs of respiratory distress at birth. All previously affected lambs from this flock bad been born dead or bad died soon after birth. The serum free-thyroxine level at 1 birth was low (9.8 pmol/l) but treatment with oral L-thyroxine resulted in an increase in thyroxine blood levels to above normal and regression of the goitre. Four weeks after withdrawal of thyroid supplementation and when the lamb was eight weeks old, the radio iodide uptake mechanism of the thyroid glands was shown to be intact but the iodide was not organified. Low normal peroxidase activity was demonstrated in fresh thyroid tissue but gel electrophoresis demonstrated an absence of normal thyroglobulin from extracts of the goitrous thyroid. This lamb and the other affected lambs of this flock have an inherited defect in thyroglobulin synthesis and it is likely that the respiratory distress was associated with the low levels of thyroid hormone affecting foetal lung development.

Journal Article↗

PubMed1986

Membrane interlocking domains in the lens.

"Ball and socket"-like membrane processes interlock fiber cells in the sheep lens cortex, but appear reduced deeper in the lens. Wheat germ agglutinin (WGA) binds preferentially to these ball and socket structures, and more weakly to other membrane regions. On protein blots, 125I WGA binds to glycoproteins with 140,000 and 32,000 apparent molecular weight, the smaller protein also binding 125I fibronectin. In two animal cataract models, the intense WGA labeling of globular bodies replaces the spotty WGA staining pattern associated with the ball and sockets in the normal lens.

Animals↗

PubMed1985

Phospholipid fatty acids in brains of normal sheep and sheep with ceroid-lipofuscinosis.

The ceroid-lipofuscinoses are a group of inherited diseases of humans and animals characterised by brain atrophy and the storage of a fluorescent lipopigment. Brain grey matter phospholipid fatty acids of diseased sheep are compared with those of normal sheep. Phosphatidylethanolamine of diseased sheep contains more 18:1(n-9) and less 22:6(n-3) than normal and their phosphatidylcholine less 16:0. Other differences are minor. All differences are in the same direction as those reported for the infantile form of human ceroid-lipofuscinosis, but are smaller. Normal sheep grey matter phosphatidylinositol contains 8.5% 20:4(n-6) and 24.6% 22:6(n-3), in contrast to 28.5 and 4.6%, respectively, in humans. The other sheep phospholipids have similar fatty acid profiles to those from humans. Apart from low levels of 20:3(n-9) and 22:3(n-9) they contain no additional non-essential fatty acid derived species. No sign of essential fatty acid deficiency occurs in either diseased or normal sheep. It is concluded that sheep must conserve their restricted essential fatty acid supply for structural functions, and that an abnormality in fatty acid metabolism is not primarily involved in the pathogenesis of ceroid-lipofuscinosis. The results also call into question the primary role of peroxidation of polyunsaturated fatty acids in lipopigment formation in this disease.

Animals↗

PubMed1984

Separation of some neutral lipids by normal-phase high-performance liquid chromatography on a cyanopropyl column: ubiquinone, dolichol, and cholesterol levels in sheep liver.

The normal-phase high-performance liquid chromatographic separation of neutral lipids into molecular classes was carried out on a cyanopropyl (CN) column eluted with isopropanol in hexane. Cholesteryl, retinyl, and dolichyl esters, triglycerides and vitamin E, ubiquinone, dolichol, phytol, and cholesterol eluted as separate peaks with 0.05% isopropanol in hexane. Cholesterol, retinol, diglyceride, and monoglyceride eluted as separate peaks with 0.75% isopropanol in hexane. These separations could not be achieved on a silica gel column. The method was used to assay sheep liver ubiquinone, dolichol, and cholesterol levels, that were determined as 77, 108 and 1864 micrograms/g wet wt, respectively.

Animals↗

PubMed1984

Ceroid-lipofuscinosis (Batten's disease). Sequential electrophysiologic and pathologic changes in the retina of the ovine model.

The sequential electrophysiologic and pathologic changes in the retina in ceroid-lipofuscinosis were recorded in a time-course study in the ovine model. Over a relatively short period in the course of the disease, a severe reduction in both rod and cone b-wave amplitudes developed with rod b-wave changes preceeding those of cones. These changes paralleled a similar loss of rod and cone photoreceptor cells. In affected retinas, outer segments appeared shorter than normal. By 84 weeks of age, the outer nuclear layer was reduced to the width of a single nucleus. In addition to these changes, electronmicroscopy showed the formation of abnormal dystrophic rod and cone outer segments in photoreceptor cells. Most cells in the retina showed the accumulation of a fluorescent lipopigment, this being most prominent in ganglion cells. Ultrastructural studies showed them to be made of electron dense granular material and a variety of membranous and tubular arrays.

Age Factors↗

PubMed1983

The catabolism of mammalian glycoproteins. Comparison of the storage products in bovine, feline and human mannosidosis.

Analysis of the neutral urinary oligosaccharides in bovine, feline and human mannosidosis by thin-layer and gel-permeation chromatography has shown that the patterns of stored oligosaccharides in the three species are different. In bovine and feline mannosidosis the most abundant urinary oligosaccharide is also the most abundant in the tissues of each species. The predominant oligosaccharides were purified by a combination of gel-filtration, ion-exchange and thin-layer chromatography and shown to contain only mannose and N-acetylglucosamine by g.l.c. and g.l.c.--mass spectrometry. The probable composition and size of each oligosaccharide were predicted from its chromatographic properties, sugar composition and the known structure of asparagine-linked oligosaccharides. The bovine and feline oligosaccharides belonged to a homologous series of general composition Mann (GlcNAc)2, whereas the human oligosaccharides belong to a different series, MannGlcNAc. These structures suggest that lysosomal endohexosaminidase is not present in bovine and feline tissues. The predominant feline storage product, Man3(GlcNAc)2, was the expected storage product from the catabolism of complex asparagine-linked glycans. In contrast, the predominant bovine oligosaccharide, Man2(GlcNAc)2, probably lacks one of the alpha-linked mannose residues in the core region. A similar situation occurs in human mannosidosis. It is predicted that in these species either that the residual mutant alpha-D-mannosidase retains activity towards one of the core alpha-linked mannose residues or that another form of lysosomal alpha-D-mannosidase that is unaffected in these disorders occurs. It is concluded that the differences in storage products are due to differences in the catabolic pathways of glycoproteins among the species.

Animals↗

PubMed1983

Small intestinal carcinoma in cattle.

The pathology of carcinoma of the small intestine in six adult cattle is described. The lesions were similar to those of carcinoma of the small intestine of sheep although a slightly less fibrous stroma and a greater tendency to metastasise was observed. Initial findings suggest the disease resembles small-intestinal carcinoma of sheep and cola-rectal carcinomas of man in having a higher prevalence in New Zealand than most other countries. This highlights the need to investigate common environmental factors which may be responsible for the induction and promotion of these tumours in both grazing ruminants and man.

Journal Article↗