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R L Hamilton

Publications and source records attributed to R L Hamilton.

144 records · Page 8Linked to original sources

Characterization of remnants produced during the metabolism of triglyceride-rich lipoproteins of blood plasma and intestinal lymph in the rat.

The metabolism of intravenously injected large and small chylomicrons from intestinal lymph and of very low density lipoproteins from blood plasma was studied in functionally eviscerated "supradiaphragmetic" rats. For studies with lymph lipoproteins, recipient animals were injected with 4-amino-pyrazolopyrimidine 18 h before injection of lipoprotein to prevent secretion of very low density lipoproteins into their blood plasma. In all cases, most of the triglycerides (labeled with 14C) were rapidly metabolized, whereas cholesteryl esters (labeled with 3H) persisted in the blood. Most of the cholesteryl esters remained in smaller "remnant" lipoproteins, less dense that 1.006, which retained an apparently spherical shape, as determined by electron microscopy of negatively stained preparations. Whereas the diameters and chemical compositions of large chylomicrons were substantially different from those of small chylomicrons and very low density lipoproteins, all remnants were similar in these respects. Average remnant diameters were 400-600 A and remnants were enriched in cholesteryl esters and in protein insoluble in tetramethylurea. In addition to triglycerides, remnants were depleted of phospholiarticle size, the composition of remnants, like that of their precursors, was consistent with the "pseudomicellar" model of lipoproteins, in which a core of nonpolar lipids is covered by a monolayer of polar lipids and protein. These results domonstrate the fundamental similarity of the initial step in the metabolism of triglyceride-rich lipoproteins from intestinal mucosa and liver and show that loss of triglycerides from the core of the particles is accompanied by removal of polar components from the surface.

Animals↗

Apoprotein composition of very low density lipoproteins of human serum.

Methods for quantitation of the major apoproteins of human serum very low density lipoprotein have been developed employing tetramethylurea, which delipidates the lipoprotein and selectively precipitates apolipoprotein B. Six soluble apoproteins are separated by electrophoresis in polyacrylamide gel. One of these is a previously unrecognized species of R-alanine (R4-alanine), more anionic than the R3-alanine polypeptide. Conditions of staining have been found which yield reproducibly linear chromogenic response with native lipoprotein and with each purified apoprotein. Recovery of protein in the seven species measured accounts for over 97% of the total in the very low density lipoprotein of normolipidemic individuals and in most samples from individuals with endogenous hyperlipemia. The mean content of apolipoprotein B in 43 samples from normolipidemic subjects was 36.9(+/-1.2 SEM)% of total protein, The distribution of the major soluble apoproteins as mean (+/-SEM) percentage of the soluble fraction was : R-serine, 5.3+/-o.5; arginine-rich, 20.6+/-1.0; R-glutamic, 10.6+/-0.4; R2-alanine, 28.3+/-0.7; R3-alanine, 26.9+/-0.5; and R4-alanine, 8.0+/-0.5. Distribution of the apoproteins was a function of particle diameter of very low density lipoprotein in fractions separated by gel permeation chromatography and by density gradient ultracentrifugation. In fractions below 700-800 A, apolipoprotein B comprised an increasing percentage of the total protein with decreasing particle diameter. Among the soluble proteins the percentage of the arginine-rich and R-serine polypeptides increased and that of the R-glutamic polypeptide declined progressively with decreasing particle size. Apoprotein distribution was similar in fractions of similar particle size from normolipidemic and hyperlipemic subjects with the exception that all fractions from the hyperlipemic subjects contained more R-serine and some, more arginine rich polypeptide. Even in the absence of chylomicrons, the distribution of soluble apoproteins in particles of diameters greater than 700-800 A was usually similar to that of the smallest particles. This suggests that the largest particles may include products of the partial catabolism of chylomicrons.

Adult↗

A simple and inexpensive membrane "lung" for small organ perfusion.

A disposable coil of thin-walled Silastic tubing, permeable to oxygen and carbon dioxide, functioned as a "lung" for perfusion of isolated rat liver. This "lung," which can be easily assembled from laboratory supplies, has a number of advantages over devices that utilize large air-liquid interfaces for gas exchange. Rates of hepatic lipoprotein triglyceride secretion, comparable to those obtained with more complex oxygenation systems, are achieved with this simple device.

Animals↗

Bacteriophage typing of clinically isolated Serratia marcescens.

A bacteriophage-typing scheme for the differentiation and classification of clinically isolated strains of Serratia marcescens was developed. Thirty-four Serratia bacteriophages were isolated from sewage and used to type 185 of 204 isolates (90.6%) of S. marcescens into 23 bacteriophage groups representing 71 types. Different bacteriophage types occurred at different intervals, suggesting that particular strains of S. marcescens are found at certain times. A correlation was found between inositol fermentation and bacteriophage type and between susceptibility to carbenicillin and bacteriophage type. However, there was no relationship between source of isolate and bacteriophage type. Bacteriophage typing of S. marcescens should provide a system which will aid in determining the origin of nosocomial Serratia infections.

Bacteriophage Typing↗

Cholestasis: lamellar structure of the abnormal human serum lipoprotein.

An abnormal lipoprotein was visualized directly in serum by electron microscopy of preparations negatively stained with potassium phosphotungstate. It appears as a unique disk-shaped particle with major axis measuring 400 to 600 angstroms and minor axis measuring about 100 angstroms. Chemical analysis, viscosity measurements, and x-ray diffraction analysis of purified preparations indicate that the particle, consisting of a one-to-one molar mixture of cholesterol and choline phosphatides associated with a small amount of protein, is a flattened vesicle, the wall of which is a continuous lipid bilayer.

Blood Protein Disorders↗

Isolation of a Golgi apparatus-rich fraction from rat liver. I. Method and morphology.

Golgi apparatus were released without fixatives from rat hepatocytes by gentle homogenization, concentrated by differential centrifugation, and purified by sucrose gradient centrifugation. Examination of sections of purified fractions by electron microscopy showed fields of morphologically intact units of Golgi apparatus consisting of stacks of parallel flattened cisternae, secretory vesicles, and small vesicular profiles. Negative staining of unfixed pellets revealed a complex network of anastomotic tubules continuous with platelike structures and secretory vesicles. These structures corresponded, respectively, to the small vesicular profiles and parallel flattened cisternae with attached secretory vesicles of sectioned material. Small fragments of granular endoplasmic reticulum were often closely associated with the peripheral tubules, suggesting sites of continuity in intact hepatocytes.

Animals↗

Characterization of lipoprotein particles isolated from the Golgi apparatus of rat liver.

It has been proposed that particles within tubules and vesicles of the Golgi apparatus of liver cells are precursors of very low density lipoproteins in blood plasma. To characterize these particles we isolated a cell fraction rich in Golgi apparatus and associated particles from rat liver in quantities sufficient for analysis. Particles freed from the membranes of the Golgi apparatus and floated at d = 1.006 were studied by chemical analysis, immunodiffusion, and paper electrophoresis. The lipid composition of the Golgi particles was similar to that of very low density lipoproteins from the same rats. The protein content was about 10% of dry weight for both the Golgi particles and plasma very low density lipoproteins. The Golgi particles formed lines of identity with plasma very low density lipoproteins during immunodiffusion against antiserum to plasma very low density lipoproteins. On paper electrophoresis, however, many Golgi particles remained near the origin, with only a few migrating to the pre-beta position. It was concluded that the lipoproteins in the Golgi apparatus are the precursors of plasma very low density lipoproteins.

Animals↗

Hepatocytic lipoprotein receptors and intracellular lipoprotein catabolism.

Hepatocytes, as the major site of synthesis and terminal catabolism of plasma lipoproteins, exert the major regulatory influence on the concentration of atherogenic lipoproteins in blood plasma and may thereby influence the rate of atherogenesis. The LDL receptor on the microvillous sinusoidal surface of hepatocytes mediates the catabolism of remnants of triglyceride-rich lipoproteins and LDL. Binding of VLDL remnants to the receptor, mediated by apo E, is of very high affinity and presumably multivalent, whereas binding of LDL, mediated by apo B-100, is monovalent and of lower affinity, accounting for the much longer residence time of the latter in the blood. The magnitude of the influx of lipoprotein particles into hepatocytic endosomal compartments dwarfs that of other macromolecules undergoing receptor-mediated endocytosis and terminal catabolism in lysosomes of these cells. The intracellular compartments and processing steps in hepatocytic lipoprotein uptake and degradation are essentially the same as those described for other ligands in the liver and other cells. Receptors with bound lipoproteins migrate into coated pits which become coated vesicles. These vesicles uncoat and fuse to form CURL vesicles and tubules near the cell surface where most receptors are recycled, presumably via receptor-rich appendages that become separated from the vesicles. CURL vesicles become mature MVBs as they migrate to the Golgi/bile canalicular pole of hepatocytes, where they fuse with putative Golgi-derived primary lysosomes and are transformed into heterophagic secondary lysosomes. MVBs also contain a receptor-rich appendage that may recycle some receptors directly to the cell surface or through adjacent Golgi compartments. Dilated ends of trans-Golgi cisternae contain nascent VLDL undergoing packaging for secretion following their synthesis and assembly in the endoplasmic reticulum. Because these "forming secretory vesicles" resemble remnant-filled MVBs, occur in a similar location in the Golgi area of hepatocytes and coisolate in centrifugal fractions of liver homogenates, there has been considerable confusion about the identity of these compartments. With the aid of specific endocytic and exocytic markers, highly purified and morphologically intact endosomal and Golgi compartments can now be obtained from rat liver homogenates. The availability of these and similar fractions of defined purity should facilitate investigation of the hepatocytic processing of endocytosed and secreted macromolecules. Although chylomicron remnants are also taken up by receptor-mediated endocytosis, the nature of the hepatocytic remnant receptor remains elusive.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Concentration and composition of lipoproteins in blood plasma of the WHHL rabbit. An animal model of human familial hypercholesterolemia.

Lipoproteins in blood plasma have been quantified and characterized in homozygous Watanabe-heritable hyperlipidemic (WHHL) rabbits, an animal model of human familial hypercholesterolemia. Like homozygous human hypercholesterolemics, WHHL rabbits have a severe deficiency of low density lipoprotein (LDL) receptors, a prolonged residence time for LDL, and an increased absolute rate of LDL catabolism. Although lipoproteins containing apolipoprotein B in WHHL rabbits are enriched in cholesteryl esters, their LDL as well as intermediate density lipoproteins (IDL) and very low density lipoproteins (VLDL) also contain a substantial amount of triglycerides and they consistently exhibit hypertriglyceridemia as well as hypercholesterolemia. The cholesteryl esters accumulating in lipoproteins of WHHL rabbits are rich in cholesteryl linoleate and appear to be produced almost exclusively by lecithin-cholesterol acyltransferase. Levels of apolipoprotein B-100 are elevated in VLDL and IDL as well as in LDL of WHHL rabbits and only trace amounts of apolipoprotein B-48 are present. Plasma levels of apolipoprotein E are also substantially increased, and VLDL and IDL are enriched in this protein. The accumulation of lipoproteins with the expected characteristics of remnants of hepatogenous triglyceride-rich lipoproteins contrasts with the efficient hepatic clearance of chylomicron remnants in WHHL rabbits.

Animals↗

Remnants of lipoproteins of intestinal and hepatic origin in familial dysbetalipoproteinemia.

We used the low molecular weight form of apolipoprotein B (B-48) as a marker for the identification of remnant particles formed from chylomicrons in the plasma of patients with familial dysbetalipoproteinemia. In the serum of patients fasted 14 hours, the d less than 1.006 g/cm3 lipoproteins of prebeta mobility, separated by starch block electrophoresis, contained only the primary hepatogenous species of apolipoprotein B (B-100), and their lipid composition resembled that of normal prebeta very low density lipoproteins. In contrast, the fraction with beta mobility contained both the B-48 and B-100 proteins; the B-48 protein was found primarily among the largest particles. All fractions of beta mobility were greatly enriched with cholesteryl esters. The beta fraction thus contains remnant particles which appear to originate both from chylomicrons and hepatogenous very low density lipoproteins. It appears that these remnant particles share a common removal mechanism which is impaired in familial dysbetalipoproteinemia.

Adult↗

[Recent advances in the neuropathological evaluation of Alzheimer's disease: the importance of alpha-synuclein].

INTRODUCTION AND METHOD: The uncertainty regarding the nosology and significance of Lewy bodies (LB) in dementia is still very present in current research. It is an acknowledged fact that an important number of cases of Alzheimer s disease (AD) also present LB. These can be detected by a synuclein immunohistochemistry in most of the sporadic cases of AD and is most frequently detected in the amygdala. CONCLUSIONS: Although the role of LB and the filaments of Lewy in the pathogenesis of dementia has still to be determined, it is clear that alphya-synuclein immunohistochemistry represents an important advance in the evaluation of the neuropathological changes in AD and in associated dementias.

Alzheimer Disease↗

[The other dementias: the neuropathology of the non-Alzheimer's disease dementias].

Alzheimer's disease (AD) is one of the most common causes of dementia, but requires neuropathological verification for a definitive diagnosis because there are a number of other neurodegenerative diseases that may present with dementia. Some of these disorders have considerable overlap both clinically and pathologically with AD, while others have distinct clinical and pathological profiles. Vascular dementia and dementia with Lewy bodies (DLB) have the greatest overlap with AD and considerable controversy still surrounds the exact contribution of the non AD pathology to the dementia syndrome. The frontotemporal dementias are loosely united by clinical presentation, but are pathologically heterogeneous and include Pick's disease, dementia lacking distinctive histopathology, motor neuron disease inclusion dementia and corticobasal degeneration (CBD). CBD can be difficult to distinguish from progressive supranuclear palsy, especially when the latter lacks the distinctive gaze palsy. Finally, Creutzfeldt Jakob disease (CJD) may be difficult to distinguish from AD when the symptoms progress at an atypically slow pace. Recently, a new variant of CJD (vCJD) that has been linked to bovine spongiform encephalopathy ('mad cow' disease) has heightened awareness of these prion protein disorders. The neuropathological criteria for the diagnosis of these non AD dementia disorders will be reviewed.

Aged↗

Herpes simplex virus brainstem encephalitis in an AIDS patient.

We report a case of a virulent, atypical herpes simplex infection in the brainstem of a patient with Acquired Immune Deficiency Syndrome (AIDS) which was characterized by demyelination and oligodendroglial tropism. At autopsy the brainstem showed demyelination. Immunocytochemistry, in situ hybridization, and electron microscopy confirmed the presence of herpes simplex virus (HSV). Viral cultures demonstrated HSV type 1. Neuroinvasiveness and neurovirulence were studied by intraperitoneal inoculation of susceptible mouse strains (A/J and Balb/cByJ) with different viral titers. The LD50 of the clinical isolate was 5 orders of magnitude greater than the LD50 of a laboratory HSV strain (HSV type 1 KOS). The brains of the mice inoculated with the clinical isolate showed brainstem and cerebellar demyelination.

AIDS-Related Opportunistic Infections↗