scholarly journals Synthesis of lipoprotein lipase in the liver of newborn rats and localization of the enzyme by immunofluorescence

1988 ◽  
Vol 249 (2) ◽  
pp. 549-556 ◽  
Author(s):  
S Vilaró ◽  
M Llobera ◽  
G Bengtsson-Olivecrona ◽  
T Olivecrona
Keyword(s):  
Lipoprotein Lipase ◽  
Gel Electrophoresis ◽  
Time Course ◽  
Polyacrylamide Gel Electrophoresis ◽  
Lipoprotein Metabolism ◽  
The Body ◽  
Newborn Rats ◽  
Adult Rats ◽  
Similar Time ◽  
Extrahepatic Tissues

In newborn rats, lipoprotein lipase (LPL) activity was higher in the liver than in several other tissues, such as heart, diaphragm or lungs, and accounted for about 3% of total LPL activity in the body. There was no significant correlation between LPL activity in liver and in plasma. Thus transport of the enzyme from extrahepatic tissues was probably not the major source of LPL in liver. To study LPL biosynthesis directly, newborn rats were injected intraperitoneally with [35S]methionine, and LPL was isolated by immunoprecipitation and separation by SDS/polyacrylamide-gel electrophoresis. Radioactivity in LPL increased with a similar time course in all tissues studied, including the liver. Substantial synthesis of LPL was also demonstrated in isolated perfused livers from newborn rats, whereas synthesis was low in livers from adult rats. There was strong LPL immunofluorescence in livers from newborn rats, mainly within sinusoids and along the walls of larger vessels. This labelling disappeared after perfusion with heparin, which indicates that much of the enzyme is in contact with blood and can take part in lipoprotein metabolism.

scholarly journals Body-Surface Compounds in Buckfast and Caucasian Honey Bee Workers (Apis Mellifera)

2014 ◽  
Vol 58 (1) ◽  
pp. 5-15 ◽  
Author(s):  
Aneta Strachecka ◽  
Grzegorz Borsuk ◽  
Jerzy Paleolog ◽  
Krzysztof Olszewski ◽  
Milena Bajda ◽  
...  
Keyword(s):  
Protease Inhibitor ◽  
Body Surface ◽  
Gel Electrophoresis ◽  
Protein Concentration ◽  
Polyacrylamide Gel Electrophoresis ◽  
Chemical Compounds ◽  
The Body ◽  
Surface Chemical ◽  
Surface Compounds ◽  
Honey Bee Workers

Abstract Body-surface chemical compounds were studied in 1-day-old nest workers and foragers both in Buckfast and Caucasian bees. The workers of these two age-castes were sampled twice in each of two consecutive years. Body-surface lipids were determined by means of gas chromatography, with a GCQ mass spectrometer. Protein concentrations and activities on the body surface were examined in bee cuticle rinsings obtained from worker bees according to the methods of Lowry, of Anson, and of Lee and Lin. Protease and protease inhibitor activities were determined. Polyacrylamide gel electrophoresis was performed. Caucasian bees, particularly foragers, had more lipids, but Buckfast bees (two age-castes) had more proteins on their body surfaces. A total of 17 alkane types (C17 - C33), 13 alkene types (C21 - C33), 21 esters (C12 - C32), and a phenol (C14) were detected in both races. Alkene C33 was detected only in Caucasian bees. More alkanes, esters, and phenols were found in Caucasian 1-day-old nest workers and foragers than in these age-castes of Buckfast bees. The protein concentration and protease inhibitor activities were lower in Caucasian bees that had higher protease activities. These values corresponded with specific numbers and widths of the electrophoretic bands.

scholarly journals INTERACTIONS BETWEEN SEX-TRANSFORMATION MUTANTS OF DROSOPHILA MELANOGASTER. I. HEMOLYMPH VITELLOGENINS AND GONAD MORPHOLOGY

Genetics ◽  
1981 ◽  
Vol 99 (3-4) ◽  
pp. 429-441
Author(s):  
T Ota ◽  
A Fukunaga ◽  
M Kawabe ◽  
K Oishi
Keyword(s):  
Gel Electrophoresis ◽  
Fat Body ◽  
Biochemical Marker ◽  
Polyacrylamide Gel Electrophoresis ◽  
Body Cavity ◽  
The Body ◽  
The Other ◽  
Sexually Dimorphic ◽  
Gonad Morphology ◽  
Sex Transformation

ABSTRACT In Drosophila, vitellogenins (yolk protein precursors) are synthesized by the female fat body, secreted into the hemolymph and subsequently taken up by the developing oocytes. The male fat body, on the other hand, does not do this even when immature ovaries are transplanted into the body cavity and grow. Thus, the hemolymph vitellogenins serve as an easily detectable sexually dimorphic biochemical marker.——We have examined hemolymph vitellcgenins by SDS polyacrylamide gel electrophoresis in flies carrying various sex-transformation mutants (dsx, tra, tra-2 and tra-20TF) singly and in all possible combinations. Chromosomal females homozygous for tra or tra-2 have no detectable hemolymph vitellogenins, while those homozygous for tra-20TR exhibit appreciable levels of these proteins. Flies homozygous for dsz, bothX / X and X / Y, have hemolymph vitellogenins, although the amount is consistently smaller in the latter. Indeed, X / Y; dsx/dsx is the only genotype in which hemolymph vitellogenins are detected in the X/Y flies. A clear hierarchy of epistasis exists among these sex-transformation mutants when they are examined in various combinations: dsx > tra, tra-2 > tra-20TF. Moreover, an interaction between tra-20TF and tra was seen in these experiments: X / X; tra-20TF/tra-20TF flies show the presence of only a trace of hemolymph vitellogenins when they are made heterozygous for tra. These results, combined with observations on gonad morphology, are discussed with respect to the BAKERa nd RIDGE (1980) hypothesis of sex determination.

scholarly journals Use of immuno-blot techniques to discriminate between the glutathione S-transferase Yf, Yk, Ya, Yn/Yb and Yc subunits and to study their distribution in extrahepatic tissues. Evidence for three immunochemically distinct groups of transferase in the rat

1986 ◽  
Vol 233 (3) ◽  
pp. 779-788 ◽  
Author(s):  
J D Hayes ◽  
T J Mantle
Keyword(s):  
Affinity Chromatography ◽  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Glutathione S Transferase ◽  
Specific Expression ◽  
Glutathione S Transferases ◽  
Extrahepatic Tissues ◽  
High Concentrations

The glutathione S-transferases are dimeric enzymes whose subunits can be defined by their mobility during sodium dodecyl sulphate/polyacrylamide-gel electrophoresis as Yf (Mr 24,500), Yk (Mr 25,000), Ya (Mr 25,500), Yn (Mr 26,500), Yb1 (Mr 27,000), Yb2 (Mr 27,000) and Yc (Mr 28,500) [Hayes (1986) Biochem. J. 233, 789-798]. Antisera were raised against each of these subunits and their specificities assessed by immuno-blotting. The transferases in extrahepatic tissues were purified by using, sequentially, S-hexylglutathione and glutathione affinity chromatography. Immune-blotting was employed to identify individual transferase polypeptides in the enzyme pools from various organs. The immuno-blots showed marked tissue-specific expression of transferase subunits. In contrast with other subunits, the Yk subunit showed poor affinity for S-hexylglutathione-Sepharose 6B in all tissues examined, and subsequent use of glutathione and glutathione affinity chromatography. Immuno-blotting was employed to identify a new cytosolic polypeptide, or polypeptides, immunochemically related to the Yk subunit but with an electrophoretic mobility similar to that of the Yc subunit; high concentrations of the new polypeptide(s) are present in colon, an organ that lacks Yc.

Determination of the proportion of κ-casein hydrolysed by rennet on coagulation of skim-milk

1980 ◽  
Vol 47 (3) ◽  
pp. 351-358 ◽  
Author(s):  
Brian Chaplin ◽  
Margaret L. Green
Keyword(s):  
Quantitative Determination ◽  
Gel Electrophoresis ◽  
Time Course ◽  
Polyacrylamide Gel Electrophoresis ◽  
Skim Milk ◽  
Casein Micelle ◽  
Clotting Time ◽  
Rate Of Hydrolysis ◽  
Hydrolysis Of

SummaryA method has been developed for quantitative determination of para-κ-casein, involving spectrophotometric scanning of stained protein bands following polyacrylamide gel electrophoresis. The rate of hydrolysis of κ-casein in skim-milk at pH 6·6 and 30 °C was compared with that in EDTA-treated skim-milk under the same conditions. This showed that at the visually observed clotting time, at least 90% of the total κ-casein in milk had been hydrolysed. The time course of the reaction was consistent with all the κ-casein molecules being hydrolysed with the same efficiency. The results strongly suggest that essentially all of the κ-casein in milk is equally accessible to rennet action. This is consistent with the casein micelle being porous, or having all the κ-casein on the surface.

Proteinases secreted by Fasciola hepatica: time course of the inhibitory effect of serum from experimentally infected rabbits demonstrated by gelatin-substrate polyacrylamide gel electrophoresis

1997 ◽  
Vol 71 (4) ◽  
pp. 333-338 ◽  
Author(s):  
L. Piacenza ◽  
D. Acosta ◽  
A. Dowd ◽  
S. McGonicle ◽  
J. Dalton ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Neutralizing Antibodies ◽  
Time Course ◽  
Fasciola Hepatica ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Group 4 ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

AbstractFasciola hepatica secretes proteolytic enzymes to aid it to penetrate and migrate through the host tissues. Two of these proteinases have been purified and shown to be cathepsin L-like, and are termed, CL1 (27.5 kD) and CL2 (29 kD). The immunogenicity of these proteinases was investigated over the course of an experimental infection and following drug treatment. Four groups of rabbits were studied: group 1: orally infected with 50 metacercariae; group 2: infected and treated 8 weeks after infection; group 3: infected, treated at week 8 and reinfected at week 13 and group 4: non-infected control group. Sera were collected weekly from each group until week 20 postinfection. CL1 and CL2 were incubated with the different sera and then analysed by gelatin substrate polyacrylamide gel electrophoresis (GS-PAGE). Analysis of groups 1, 2 and 3 showed that CL1 and CL2 neutralizing antibodies appear at week 5 post-infection. In group 1, these remained throughout the 20 weeks of infection. In group 2, neutralizing antibodies disappeared at week 13, that is, 5 weeks after anti-Fasciola treatment. In group 3, CL1 and CL2 neutralizing antibodies disappeared at week 13 but reappeared by week 15, that is 2 weeks after reinfection. Pooled sera from group 4, showed no inhibitory capacity. ELISA results using CL1 and CL2 as antigen, correlate very well with the inhibitiory time course observed by GS-PAGE. These results suggest that purified cathepsin Ls are antigenic molecules recognized early in the infective process and capable of inducing a specific humoral response, strong enough to neutralize, at least partially, their enzymatic activity.

scholarly journals Purification and characterization of rat adipose tissue lipoprotein lipase

1982 ◽  
Vol 207 (3) ◽  
pp. 485-495 ◽  
Author(s):  
S M Parkin ◽  
B K Speake ◽  
D S Robinson
Keyword(s):  
Adipose Tissue ◽  
Affinity Chromatography ◽  
Lipoprotein Lipase ◽  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Antithrombin Iii ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Sepharose 4B

Lipoprotein lipase (EC 3.1.1.34) extracted from adipose tissue of glucose-fed rats with 5 mM-sodium barbital, pH 7.5, containing 20% (v/v) glycerol and 0.1% (v/v) Triton X-100, was partially purified by affinity chromatography on heparin linked to Sepharose 4B. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of the partially purified enzyme preparation revealed the presence of two major Coomassie-staining bands (mol.wts. 62 000 and 56 000) as well as a number of minor bands. Treatment of partially purified enzyme with [1,3-3H]di-isopropyl fluorophosphate resulted in the incorporation of radiolabel into the band of mol.wt. 56 000, but not into the band of mol.wt. 62 000. Both the amount of the 56 000-mol.wt. polypeptide and the incorporation of [1,3-3H]di-isopropyl fluorophosphate into this band were greatly reduced in the enzyme preparations isolated from adipose tissue of 48 h-starved rats. whereas the amount of the 62 000-mol.wt. polypeptide was unaffected by starvation. Purification of lipoprotein lipase from adipose tissue of glucose-fed rats was also carried out using affinity chromatography on Sepharose 4B linked to heparin with low affinity for antithrombin-III. This procedure resulted in the presence of a single band of mol.wt. 56 000 on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. These results suggest that the polypeptide of mol.wt. 56 000 corresponds to the subunit of lipoprotein lipase, whereas the 62 000-mol.wt. polypeptide probably represents antithrombin-III.

scholarly journals The characterization of the non-histone chromosomal proteins of the main classes of nuclei from rat brain fractionated by zonal centrifugation

1979 ◽  
Vol 177 (1) ◽  
pp. 331-346 ◽  
Author(s):  
Sonia G. Tsitilou ◽  
David Cox ◽  
Anthony P. Mathias ◽  
David Ridge
Keyword(s):  
Rat Brain ◽  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Brain Cells ◽  
Adult Rats ◽  
Chromosomal Proteins ◽  
Neuronal Nuclei ◽  
Infant Rats

1. Non-histone chromosomal proteins were isolated from the cell nuclei of whole rat brain and nuclei from different types of brain cells. 2. Brain nuclei were fractionated by zonal centrifugation into five zones deriving from five main categories of brain cells. These are the neuronals, astrocytes I, astrocytes II, oligodendrocytes I and oligodendrocytes II. 3. The non-histone chromosomal proteins were analysed by (a) sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, (b) electrofocusing electrophoresis and (c) two-dimensional electrophoresis. The results of this analysis showed a limited specific pattern of non-histone chromosomal proteins from the different classes of nuclei. Differences were found to exist between the proteins from neuronal and glial nuclei. In particular one polypeptide band with mol.wt. 10000 and pI8.5 was found to be present in the non-histone protein fractions of neuronal nuclei, and absent from the corresponding fractions of nearly all the other classes of nuclei. 4. Two other classes of nuclear proteins, buffered-saline-soluble and 0.35m-NaCl-soluble, were analysed by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis along with the non-histone chromosomal. The similarities and differences among these groups of proteins are discussed. 5. The patterns of non-histone chromosomal proteins during development were investigated by studying them in two age groups of animals: in infant rats (10 days old) and adult rats. The polypeptide that was found to be specific for the proteins of neuronal nuclei of adult rats is present in all the classes of nuclei of infant rats.

scholarly journals Properties of salt-resistant lipase and lipoprotein lipase purified from human post-heparin plasma

1979 ◽  
Vol 179 (3) ◽  
pp. 555-559 ◽  
Author(s):  
A M Ostlund-Lindqvist
Keyword(s):  
Amino Acid ◽  
Amino Acid Composition ◽  
Lipoprotein Lipase ◽  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Bovine Milk ◽  
Molecular Weights ◽  
Heparin Plasma

Lipoprotein lipase and salt-resistant lipase were isolated from human post-heparin plasma. The proteins of human post-plasma lipoprotein lipase and salt-resistant lipase were identified and demonstrated to be immunologically different. Significant differences between the two enzymes in their relative amino acid composition were demonstrated, which indicates that the two enzymes are different proteins. When analysed by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, the enzymes seemed to have monomer molecular weights similar to that of lipoprotein lipase purified from bovine milk.

Time course of adaptation to hypoxia in newborn rats

1988 ◽  
Vol 66 (1) ◽  
pp. 152-158 ◽  
Author(s):  
Tiziana Piazza ◽  
Anne Marie Lauzon ◽  
Jacopo P. Mortola
Keyword(s):  
Body Weight ◽  
Time Course ◽  
Breathing Pattern ◽  
The Body ◽  
Adaptation To Hypoxia ◽  
Newborn Rats ◽  
Time Intervals ◽  
Lung Weight ◽  
Heart Mass ◽  
C Value

In newborn rats after a few minutes of hypoxia, ventilation is similar to the normoxic value. Nevertheless, after a few days in hypoxia, newborn rats have a sustained hyperventilation. In this study we examined the time course of the newborn rat's adaptation to hypoxia. Measurements of body size, hematocrit, lung and heart mass, and breathing pattern have been performed on newborn rats exposed to hypoxia (10% O2) for different time intervals from 4 to 60 h (hypoxic, H), and on same-age rats growing in air (controls, C). Ventilation measured by flow plethysmography was increased in H rats above the C value from about 8 h; this was due to a higher breathing rate and, from 24 h, also to a larger tidal volume. During the early hours of hypoxia, oxygen consumption measured manometrically was about 50% of C, while after 3 days in hypoxia it was almost like the C value. These observations indicate that the lack of sustained hyperventilation, characteristic of the newborn's acute exposure to hypoxia, is an immediate but transient phenomenon that is resolved after a few hours, and suggest a tight link between metabolic and ventilatory hypoxic responses. Body weight of H rats was less than in C, owing to an immediate decrease below the prehypoxic value. Dry heart and lung weight changed in proportion with the rest of the body during the first 36–48 h of hypoxia, then they increased disproportionately more. Hence, these temporal changes suggest that the large heart and lung weight – body weight ratios of the chronic hypoxic animals result from their smaller body mass and the stimulated growth of cardiac and pulmonary tissues.

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