Gene/Protein Disease Symptom Drug Enzyme Compound
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We previously reported that the pulmonary intravascular macrophages (PIMs) of sheep, goat, and calf lung contained a heparin and a lipolytic lipase sensitive surface coat by using tannic acid as a component of paraformaldehyde-glutaraldehyde-based fixative. The implication of this sensitivity was that the surface coat was predominantly comprised of lipoprotein-like substance. In this study we report that monastral blue (MB) used as a vascular tracer interacted with the coat globules and lost its original particulate appearance. Its precise localization in the PIMs was in combination with altered macromolecules of the surface coat in the form of lipid droplets, which conformed to the conventional view of neutral lipids. In contrast, pigment particles examined in their native state resembled metallic particles as electron-dense elliptical rods. The lipid droplets were subsequently internalized through endocytic route and found their access into the lysosomal compartments of PIMs at the electron microscopic level. Lamellar bodies (LLBs) arose from the lysosomal matrix after the entry of lipid droplets in the secondary lysosomes. Acid phosphatase activity was located in secondary lysosomes as well as in endosomes. These observations suggest that coat granules of the PIMs acted as a carrier of exogenous MB particles to deliver the complex to the lysosomal compartment where partial digestion lead to the formation of lamellar bodies. The implications of MB (cationic dye) as a vascular tracer for studying phagocytic index of PIMs in the light of their coat and the rapid development of LLBs are discussed. It is proposed that MB by initially combining with the surface coat provokes mobilization of intracellular lipid pools. In this way metabolism of vasoactive lipid in the PIMs is stimulated to influence the dynamics of pulmonary circulation in the calves.
Anat Rec 1992 Oct
PMID:In vivo monastral blue-induced lamellar-bodies in lysosomes of pulmonary intravascular macrophages (PIMs) of bovine lung: implications of the surface coat. 141 8

The pulmonary intravascular macrophages (PIMs) have been described in several species of animals. This study demonstrates for the first time that the equine lung has PIMs as resident phagocytes in its microvasculature. Their salient features such as globular surface coat, structures of the endocytic pathway, and related cell organelles closely resemble those of the calf, goat, and sheep. The exquisite organization of the coat globules in the form of a linear chain was structurally similar to the lipolytic lipase and the heparin-sensitive globular coat from PIMs of calf, goat, and sheep. Monastral blue (MB) when employed as a tracer to assess the phagocytic properties of equine PIMs induced similar modification of the globules of the coat into lipid droplets, reminiscent of neutral lipids. Lipids droplets (modified coat globules) were delivered into acid phosphatase-positive endosomes and lysosomes. Concurrently, the unaltered globules of the coat, probably internalized via fluid-phase constitutive pinocytoses, followed a different endocytic pathway. Large-scale platelet uptake by the PIMs was observed with thrombocytopenia in MB-treated ponies. The possible significance of hypothetical LDL-coat and the endocytic organelles as equivalents of synthetic apparatus of vasoactive lipids in the PIMs of horse needs to be assessed in future studies.
Anat Rec 1992 Dec
PMID:Presence of pulmonary intravascular macrophages in the equine lung: some structuro-functional properties. 145 55

The effect of progressive, diabetes-associated adiposity on reproductive tract structure and function was examined in 4- to 16-week-old C57BL/KsJ, control (+/?) and diabetic (db/db) mice. Uterine and ovarian tissues were analyzed by transmission electron microscopy for ultrastructural changes associated with increased intracellular lipid accumulation. In addition, the same tissues were analyzed for changes in activity of tissue lipoprotein lipase, an enzyme that hydrolyzes lipoprotein-associated triacylglycerols and supports the cellular uptake and storage of free fatty acids. Between 8 and 16 weeks of age, intracellular lipid deposits increased dramatically in the ovarian granulosa, thecal and stromal cell populations, as well as in the uterine epithelium, of diabetic mice compared to controls. By 16 weeks of age, the lipid deposits essentially occupied the entire cytoplasmic area of both the ovarian and uterine cell types in diabetics. The basal lamina underlying the uterine epithelium was expanded in the diabetics relative to controls, and the hyperglycemic condition induced an observable increase in endometrial intercellular space that was occupied by a hyaline type of ground substance of unknown composition and origin. In association with these structural changes, both ovarian and uterine lipase activities were greatly increased in the db/db mice compared with controls. These data suggest that the structural adiposity and functional decline in reproductive tract condition of the db/db mutants are related to the enhanced cellular lipid deposition observed in this species. These changes in structural and metabolic parameters are related to the reproductive incompetence characteristic of this murine model.
Anat Rec 1986 Nov
PMID:Ultrastructural and metabolic changes associated with reproductive tract atrophy and adiposity in diabetic female mice. 378 18

Magnesium, calcium, phosphate, total protein and free fatty acid concentrations, and a lipase activity using p-nitrophenylpalmitate NP-C16) as a substrate, were determined in the serum of 15 pregnant Aberdeen Angus cows grazing a Phalaris tuberosa pasture with a tendency to induce tetany. A gradual decrease in magnesium concentrations, but no definite change in the other parameters, was observed when the results were plotted directly against the time of the experiment. When the results were plotted as a function of the stage of pregnancy or lactation of the animals, magnesium concentrations showed a more steady decrease which was associated with increases in calcium concentrations and, at the lowest magnesium concentrations, with increases in phosphate concentrations. Total proteins and free fatty acids remained fairly constant and the highest lipase activity was associated with parturition. The lowest magnesium concentrations in serum did not correspond with high levels of either lipase or free fatty acids. The hypomagnesaemia observed in this experiment was attributed to the high potassium:calcium and magnesium ratio in the pasture and to the physiological condition (pregnant and lactating) of the animals. None of the parameters evaluated in this paper would be more sensitive than serum magnesium levels for the early detection of the conditions that predispose grazing cattle to grass tetany.
Vet Rec 1984 Dec 01
PMID:Changes in blood composition of pregnant cows during the onset of hypomagnesaemia. 652 81

A trial was conducted to establish whether there would be any benefit in feeding mammalian pancreatic enzymes to farmed salmon clinically affected by pancreas disease. Pancreatic enzymes were incorporated at a rate of 4 kg/tonne of feed and fed at 0.6 to 0.9 per cent bodyweight/day for 41 days to one cage of salmon. The weights, lengths and condition factors were established for the fish before and after the feeding trial and compared with those for an adjacent cage of untreated fish. Histopathology and blood amylase and lipase activities were monitored weekly. No significant increases in weight or condition factor, and no reduction in mortalities were recorded. Histopathology confirmed the presence of pancreas disease throughout the trial and there were concurrent skeletal and cardiac myopathies. Blood amylase activities showed no significant change and were considered to be within normal ranges; blood lipase activity remained undetectable.
Vet Rec 1995 May 13
PMID:Apparent lack of response of salmon affected by pancreas disease to pancreatic enzyme replacement therapy. 767 89

Plasma lipase concentrations were determined in Atlantic salmon post smolts at weekly intervals on two farms from late June. On one farm there was a significant increase (P < 0.001) in lipase concentration which coincided with a suspicion of pancreas disease on clinical grounds. A definitive diagnosis was subsequently confirmed by histopathology. The exercise was repeated on 10 farms in the following year and the results provided additional evidence of the value of monitoring lipase concentration as an indication of pacreas disease at an earlier stage than it can be detected by clinical signs and histopathology alone.
Vet Rec 1994 Jul 30
PMID:Plasma lipase concentration as an aid to the early detection of pancreas disease in farmed Atlantic salmon (Salmo salar). 873 80

Pulmonary intravascular macrophages (PIMs) contain a unique electron-dense globular surface-coat which is sensitive to heparin treatment, halothane anesthesia, and the digestive effect of lipolytic lipase (LPL), suggesting that the coat is predominantly composed of lipoproteins. In the present study, evidence is presented that heparin, when administered intravenously in goats, potentiated both the translocation of the surface-coat into the vacuolar system and the expansion of the Golgi apparatus. Sequentially, these changes were followed by proliferation of peroxisomes in combination with peroxisomal reticulum (PR), a transient precursor of this organelle. The peroxisomes, as well as PR, reacted positively for catalase after aldehyde fixation and 3,3'-diaminobenzidine (DAB) staining. In addition to their role as phagocytes, the ultrastructural and cytochemical detection of peroxisomes suggests a functional capacity of the PIMs, which may be adaptable to the circulating level of free fatty acids (FAAs).
Anat Rec 2002 01 01
PMID:In situ heparin-induced peroxisomal reticulum and biogenesis of peroxisomes in pulmonary intravascular macrophages (PIMs) of caprine lung: an ultrastructural and cytochemical study. 1174 73

Among the various kinds of acyl donors, the 1-alkoxyvinyl esters have characteristic features, such as a high reactivity under nearly neutral conditions and the generation of neutral and volatile esters as single coproducts. Although their use in organic syntheses began in the middle of the 1950s, no significant progress has been seen. This is probably because the existing method of preparing alkoxyvinyl esters used toxic mercuric salts and was not totally applicable for those esters having functionalized acyl moieties. We have discovered that the use of a catalytic amount of the less toxic [RuCl2(p-cymene)]2 effectively accelerates the addition of carboxylic acids to ethoxyacetylene to give ethoxyvinyl esters bearing a variety of functionalized acyl groups in high yields. This discovery has opened a new avenue for developing new reactions and new synthetic methodologies based on the design and use of these acyl donors with suitable functional groups. Such examples include (i) the installation of hydrophilic acyl moieties on biologically active compounds, (ii) asymmetric Pummerer-type reactions, (iii) aromatic Pummerer-type reactions, (iv) the lipase-catalyzed desymmetrization of symmetrical 1,3-diols, and (v) lipase-catalyzed domino reactions. Future possibilities for these acyl donors are also discussed.
Chem Rec 2004
PMID:1-alkoxyvinyl esters: renaissance of half-century-old acyl donors with potential applicability. 1573 98

Carefully designed molecules that are intimately related to the reaction mechanism of enzymes are often highly selective and potent inhibitors that serve as extremely useful chemical probes for understanding the reaction mechanism and structure of enzymes. This article describes the design, synthesis, and applications of specific inhibitors of two mechanistically distinct groups of enzymes, ATP-dependent amide ligases and Ser- and Thr-hydrolases. Our strategy is based on the premise that stable analogues of the transition state (transition-state analogues) are highly potent inhibitors that serve as good mechanistic probes, and that a key structure of a good inhibitor of one enzyme is also utilized for the inhibitors of other enzymes that share the same chemistry in their catalyzed reactions, irrespective of the degree of structural similarity and evolutionary link between the enzymes. According to these principles, we designed and synthesized a series of phosphinate- and sulfoximine-based transition-state analogue inhibitors of glutathione synthetase, gamma-glutamylcysteine synthetase and asparagine synthetase. For the second group of enzymes, we synthesized a gamma-monofluorophosphono glutamate analogue for mechanism-based affinity labeling of gamma-glutamyltranspeptidase and fluorescent phosphonic acid esters for the active-site titration of lipase. These inhibitors were used successfully as ligands for detailed kinetic analyses, X-ray crystallography, and mass analysis of the enzymes to identify the key amino acid residues responsible for catalysis and substrate recognition in the transition state.
Chem Rec 2005
PMID:Enzyme inhibitors as chemical tools to study enzyme catalysis: rational design, synthesis, and applications. 1604 44

In situ adsorption, known as an in situ-roduct removal (ISPR) technique for low molecular mass bioproducts, was in this study applied to a bacterial exoenzyme proving that this method is also suitable for the separation of macromolecules like proteins. For this, adsorbent particles were added to growing cultures of Staphylococcus carnosus rec., therefore both production and adsorption occurred simultaneously in shaking flasks, stirred tank, or airlift bioreactor as the chosen types of fermenters. The exoenzyme lipase adsorbed rapidly and, after separating cells and adsorbents, desorbed in a packed bed column. Up to 85% of the produced lipase were recovered, fractions of these had been concentrated up to the factor 20 and purified up to a factor of 40 by the procedure. By using the airlift bioreactor an enhancement of biomass production was observed, but the necessity of the addition of an anti-foam reagent resulted in higher product losses in adsorption as well as in desorption. Production and adsorption kinetics have been modeled and applied to in situ-adsorption. The model was used to perform a parameter study in which the influence of biological and physical parameters as well as process parameters on discontinuous and continuous in situ-adsorption was investigated.
 ...
PMID:Process modeling of in situ-adsorption of a bacterial lipase. 1626 49


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