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Density gradient centrifugation using a performed self generated gradient of colloidal silica enabled the isolation of microscopic sheep sarcocystis cystozoites, free from heart muscle contamination. The efficiency of separation of cystozoites from residual heart muscle after digestion in pepsin and hydrochloric acid was 63 to 92 per cent. Antigens from cystozoites were used on enzyme-linked immunosorbent assays (ELISA) of plasma from six coccidia-free lambs infected once orally with 70,000 microcystic sheep sarcocystis sporocysts and for raising antisera in rabbits. Use of an anti-sheep IgM conjugate in the ELISA showed that anti-sarcocystis IgM production was transitory, appearing five to 10 days after infection, peaking in concentration at 42 days and following the peak of the acute phase of infection (32 and 33 days) in the lambs. In contrast, total anti-sarcocystis immunoglobulins, detected by ELISA, increased from five to 21 days after infection and continued to increase until the lambs were killed (the last at 81 days) and was more useful in diagnosing chronic infection. No cross reactions between microcystic sheep sarcocystis and Toxoplasma gondii or Eimeria species of sheep were observed. A peroxidase anti-peroxidase test, using rabbit anti-sarcocystis sera, detected second generation meronts and sarcocysts in fixed tissues from infected lambs making it useful for the diagnosis of acute or chronic disease post mortem.
Vet Rec 1986 Nov 29
PMID:Experimental microcyst sarcocystis infection in lambs: serology and immunohistochemistry. 309 51

Renal tubular cells and segments isolated by a trypsin, pepsin, pronase E digestion procedure were studied with scanning electron microscopy. The basal and lateral surfaces of S1, S2, S3 proximal tubular (PT) segments, descending and ascending thin limbs of Henle (TL), distal ascending thick limb of Henle, or distal straight tubule (DST) and distal convoluted tubule (DCT) segments, connecting tubules (CNT), and collecting ducts (CD) were identified and characterized. The basal processes of the S1 and S2 PT cells were fan shaped, were oriented in a circumferential direction, and terminated in microvilli at the basement membrane. S3 PT cells had microvillous basal processes mainly on the lateral edges of the cells. The basal processes of DST and DCT were similar to PT in orientation but terminated on the basement membrane with flattened, thin attachments. The long-loop descending TL and the ascending TL exhibited distinctive interdigitating cell processes. TL segments with simple contours were present in smaller numbers and were characteristic of short-loop descending limbs. CNT showed some cells with basal surfaces resembling DCT cells and others resembling CD cells. Both cortical and medullary CD segments exhibited intercalated cells with round basal contours and a sparse pattern of basal infolding clefts. The cortical CD principal cells revealed a much more elaborate mosaic of plicae, clefts, and microvilli than those of the medullary CD. These observations extend the previous knowledge gained from transmission electron microscopy and assist in the interpretation of that knowledge.
Anat Rec 1985 Oct
PMID:Scanning electron microscopy of basolateral surfaces of rat renal tubules isolated by sequential digestion. 390 17

The ultrastructure of serous cells from porcine tracheal submucosal glands was studied by conventional transmission electron microscopy (TEM), and by cytochemical methods to stain for complex carbohydrates. In tissue fixed and processed for TEM, and stained with uranyl acetate and lead citrate, the condensing granules of serous cells occasionally possessed a hexagonal and sometimes a lamellar substructure. Tissue fixed in paraformaldehyde-glutaraldehyde and stained with periodic acid-thiocarbohydrazide-silver proteinate (PTS) or with phosphotungstic acid (PTA) showed secretory granules stained for complex carbohydrates and revealed a substructure similar to that noted in the condensing granules. The dark staining substructure revealed by either the PTS or the PTA technique appeared to correspond to electron-lucent areas observed in the condensing granules by conventional TEM. The PTS staining probably demonstrated the presence of neutral glycoprotein, since the serous-cell granules did not react with a dialyzed iron stain for acidic glycoproteins. Treatment of periodic acid oxidized thin sections with pronase or pepsin prior to thiocarbohydrazide and silver proteinate treatment decreased the intensity of the PTS staining, but did not digest away any components of the granules. The substructure revealed by the carbohydrate stains may be a reflection of the mechanism of packaging or the macromolecular structure of the glycoproteins in the serous-cell granules.
Anat Rec 1982 Jul
PMID:Substructure of granules from serous cells of porcine tracheal submucosal glands. 618 18

The bronchiolar Clara cells of rats contain characteristic rod-shaped granules always surrounded by a unit membrane. These granules contain thin filaments about 9 to 10 nm in diameter lying in a pale matrix. Our morphological results suggest that the filamentary rod-shaped granules originate from the common, round-to-oval, electron-dense Clara cell granules, as we found different intermediate structures between these two kinds of granules. The electron-dense granules are digested by pepsin, whereas the filamentary rod-shaped granules are apparently not affected. The biochemical nature and the possible function of the filamentary rod-shaped granules are also discussed in relation to the secretory activity of the Clara cells.
Anat Rec 1981 Oct
PMID:Filamentary rod-shaped granules in Clara cells of rats. 679 22

The expression of the complete human gastric lipase (HGL) gene in Saceharomyces cerevisiae grown in defined medium resulted in the secretion of active recombinant HGL (rec.HGL) to levels of up to approximately 11 mg/liter. Of the total measurable HGL activity, 90% was detected by assaying intact cells, suggesting that the majority of rec.HGL produced was secreted but stayed attached to the cell wall. The remaining 10% was present in the growth medium and from this source active rec.HGL was purified 300-fold by a combination of hydrophobic interaction and ion-exchange chromatography. Rec.HGL migrated on reduced SDS-PAGE as three bands with estimated molecular masses of 47,45, and 43 kDa. All three forms cross-reacted with an antibody raised to natural HGL and their treatment with Endo H showed them to be N-linked glycosylation variants of a single polypeptide. The 47-kDa species was isolated using lentil lectin Sepharose 4B and shown to possess a specific activity comparable to that of the natural enzyme. Rec.HGL had an acid pH activity optimum using either tributyrin or olive oil as substrate and did not lose activity if incubated in the presence of pepsin at pH 2.0. These results demonstrate that HGL secreted by Saccharomyces cerevisiae retained those properties of the natural enzyme required for its use in the treatment of pancreatic insufficiency.
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PMID:The secretion of active recombinant human gastric lipase by Saccharomyces cerevisiae. 886 Jun 47

The prevalence of Sarcocystis species in muscle samples from gazelles kept as breeding groups at the King Khalid Wildlife Research Centre, Saudi Arabia, was determined by fibreoptic examination, pepsin digestion and histological techniques. No macroscopic sarcocysts were detected by fibreoptic examination, and the overall prevalence of Sarcocystis was 66 x 7 per cent by pepsin digestion, and 39 x 9 per cent by histological examination. By digestion, the tongue contained the highest density of bradyzoites in Gazella dorcas, and Gazella gazella erlangeri, the oesophagus in Gazella subgutturosa marica and skeletal muscle in Gazella gazella and Gazella thomsoni. Skeletal muscle was least affected in G dorcas, the oesophagus in G gazella, and the diaphragm in G g erlangeri, G s marica and G thomsoni. By histology, the heart contained most microcysts, except in G g erlangeri, in which the tongue was most affected. No single tissue type was therefore suitable for the diagnosis of sarcocystosis in this multispecies collection, although digestion was more sensitive in detecting infection than histology. The level of Sarcocystis infection was significantly higher in free-ranging gazelles kept in a main enclosure than in gazellas kept in breeding pens, and higher in adult gazelles than in juveniles.
Vet Rec 2000 Feb 19
PMID:Sarcocystis infections in gazelles at the King Khalid Wildlife Research Centre, Saudi Arabia. 1073 Oct 71