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Semen samples were collected at weekly intervals for six weeks from eight sexually mature beagles previously shown to produce normal ejaculates. Seminal plasma and sperm fractions were separated by centrifugation and the sodium, potassium, alanine and aspartate aminotransferases, acid and alkaline phosphatase concentrations in the two fractions determined. Regression analysis of the mean weekly values obtained from physical and biochemical examination of the ejaculates showed that sodium ion concentration was highest in seminal plasma. The highest levels of aminotransferases were found in sperm fractions. Those enzymes may be indices of abnormal or damaged spermatozoa. Acid and alkaline phosphatase activity was 100 times greater in seminal plasma than in sperm fractions. Phosphatase concentrations are likely to be dependent on prostate activity. Measurement of acid phosphatase in canine semen therefore may be a useful index of prostate function. The motility of the semen samples was independent of the potassium concentration in seminal plasma. However, there was some evidence of a correlation between sperm motility and the enzyme and sodium content of seminal plasma.
Vet Rec 1979 May 26
PMID:Biochemical observations on beagle dog semen. 47 66

The history of particle clearance was studied in lungs of mice serially sacrificed at intervals up to 14 months following single exposures to an aerosol of submicronic, particulate, iron oxide used as a similitude for atmospheric dust. Clearance was followed by light microscopy in unstained and Prussian blue stained frozen and plastic embedded sections, as well as by electron microscopy, where iron oxide can be recognized by its form. Related problems were investigated through histochemical demonstration of acid phosphatase activity in pulmonary lysosomes and Prussian blue staining of various tissues after administration of iron compounds by gastrointestinal and vascular routes. The iron particles settle extensively but not uniformly on pulmonary alveolar surfaces. Clearance is centripetal and involves two mechanisms, an extracellular mechanism fed by fluid currents sweeping across the surfcace, and cellular mechanism principally involving alveolar macrophages. In the early post exposure period both actively remove deposited particles predominantly through the pulmonary airways. By 24 hours uncleared residues have become ingested and clearance thereafter results mainly from cellular action. Macrophages enter bronchial passages where they sometimes continue to pursue normal activities. A chronic phase of clearance begins when deposited particles become sequestered in macrophages of pulmonary connective tissues. These cells are reached by several routes, not least by crossing the bronchial epithelium. Particle clearance from these macrophages is very slow, and residue-containg cells eventually congregate in lymphoid tissues surrounding major bronchi. These findings are discussed as they help to develop an overall picture of clearance from the lungs and as they bear on related topics, such as functional roles of alveolar and pulmonary connective tissue macrophages and the pathogenesis of chronic bronchial disease.
Anat Rec 1975 Mar
PMID:Pathways of clearance in mouse lungs exposed to iron oxide aerosols. 112 38

At the close of a uterine cycle, the remodelling of the endometrial stroma of the North American opossum involves removal of extracellular material by macrophages. This study provides cytochemical and ultrastructural evidence which indicates that the laden macrophages are eliminated from the endometrium through emigration across the glandular and luminal epithelia. During diestrus or the early postpartum period, the abundant uterine glands relinguish their secretory function to acquire a transient function in the transportation of emigrating stromal cells. During the first three postpartum days endometrial regression in the stroma is marked by sudden appearance of monocytes, macrophages, lymphocytes, and plasma cells. Ultrastructural and cytochemical evidence indicates that the macrophages engulf the extracellular macromolecular material which, in the opossum, consists primarily of ground substance. Macrophages filled with ingested extracellular material aggregated beneath the glandular and luminal epithelia, where they acquire an extracellular coat that resembles the material of the basal lamina elsewhere. A fibroblast-like cell closely invests the macrophage at the time the extracellular material appears. Simultaneously, the secretory glandular epithelium is being converted to a highly ciliated one. Macrophages, often accompanied by lymphocytes, acquire intraepithelial positions in the glands. From here these stromal cells gain entrance to the glandular lumens. At this time the luminal contents are rich in acid phosphatase activity which most likely reflects the high lysosomal content of the emigrating macrophages. Evidence suggests that these intraluminal macrophages and lymphocytes are swept, by the recently differentiated ciliary lining, toward the glandular orifices and into the uterine cavity. It is hypothesized that this cyclic appearance and transepithelial elimination of macrophages is a cellular mechanism for removing large amounts of extracellular material without disruption of the endometrium.
Anat Rec 1976 Jan
PMID:Cellular mechanisms involved in cyclic stromal renewal of the uterus. I. The opossum, Didelphis virginiana. 125 15

The cytochemical changes of the Golgi stacks occurring concomitantly with cell differentiation were examined in ameloblasts of developing rat molar tooth germs using osmium impregnation and cytochemistry with nicotinamide adenine dinucleotide phosphatase (NADPase), thiamine pyrophosphatase (TPPase), and acid phosphatase (Acpase). NADPase, TPPase, and Acpase activities were already present in the Golgi stacks of the inner enamel epithelial cells, the undifferentiated form of the ameloblast: NADPase activity existed in the medial Golgi cisternae, TPPase activity in the trans Golgi cisternae, and Acpase activity in almost all cisternae and strongly in the trans-most cisterna of the Golgi stack. At this stage, however, osmium deposits after impregnation were not observed in the cisterna of Golgi stacks but were present in some small vesicles. These vesicles were located throughout the cytoplasm. Osmiophilic cisternae in the Golgi stacks were apparent for the first time at the stage when the Golgi apparatus developed and migrated to the region distal to the nucleus with the progression of cell differentiation. These findings indicate that the cis subcompartment of the Golgi apparatus was incomplete in the inner enamel epithelial cells with regard to appearance of its cytochemical property, as compared with the medial and trans subcompartments. It is suggested that the cis compartment of the Golgi stack may be completed only in the last stage of the compartmentalized Golgi organization during differentiation of the ameloblast.
Anat Rec 1992 Dec
PMID:Changes of cytochemical properties in the Golgi apparatus during in vivo differentiation of the ameloblast in developing rat molar tooth germs. 145 50

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 fate of macrophage precursors residing in 14-day prenatal rat lungs was followed in organ cultures to obtain a detailed, ultrastructurally resolved picture of the sequence and timing of events accompanying their transformation into typical pulmonary macrophages. Cultures were examined at close intervals during the first day (1, 2, 3, 4, 6, 9, 12, 15, 18, and 24 hr) and at wider intervals thereafter (2, 4, 5, 7, 9, and 13 days) to yield a developmental series of cells identified as in the macrophage line based on binding of peroxidase-coupled isolectin B4 of Griffonia simplicifolia (GSA I-B4) to cell membranes and on negligible content of peroxidase-positive granules in the cytoplasm. Organ culturing stimulated virtually all precursors to develop into macrophages. GSA-positive cells in explants occurred outside vessels in pulmonary connective tissue, and at the outset none were typical macrophages: 71% were angular cells, resembling unlabeled mesenchymal cells around them, 16% were undifferentiated leukocytes, and the remainder were irregularly shaped cells with few vacuoles intermediate between the preceding and the macrophages. During the first 12 hr in culture the proportion of angular cells and leukocytes fell to zero, and that of intermediate cells first rose, then receded. In the same interval the proportion of macrophages rose to 87.5%, and by 24 hr all GSA-positive cells were typical macrophages generally engorged with phagocytosed material; about 8 hr appear necessary for converting half the population. Notable ultrastructural changes during this period of transformation involved the centrioles and cytoskeleton, reflecting enhanced cell mobility and phagocytosis. A period of maturation followed, marked by disappearance of cellular debris from phagosomes and an increased prevalence of cells with elaborate lamellipodia. This accords with earlier work showing that macrophage Fc receptor density increases sharply during the first 24 hr, but elevated levels of histochemically demonstrable acid phosphatase appear only later. Mitotic activity was conspicuous in GSA-positive cells throughout both periods. 3H-thymidine labeling indices for precursors and macrophages, determined at six intervals between 1 hr and 24 hr, remained steady at approximately 34%, whereas indices of other categories of lung cells (GSA-negative stromal cells, pleural cells, and airway epithelium) began at this level but rapidly declined, indicating that the GSA-positive cells constitute a single population distinct from others in the lungs. Macrophages found outside the lung cultures after 4-5 days qualify as a mature population, but having migrated away from direct contact with the lung stroma, they survive only a week or two and no longer divide.
Anat Rec 1992 Apr
PMID:Macrophage development: III. Transformation of pulmonary macrophages from precursors in fetal lungs and their later maturation in organ culture. 155 5

Dipeptidyl peptidase II (DPP II), E.C. 3.4.14.2, a serine class endopeptidase, is widely used as a lysosomal marker in cytochemical studies. To date most ultrastructural studies of ameloblasts use the presence of acid phosphatase activity to identify cellular organelles to be lysosomal. Using decalcified rat mandibles, with kidney tissue as a positive control, DPP II activity, was assessed with specific substrate Lysyl-alanine-4-methoxy-2-naphthylamide in ameloblasts at an ultrastructural level. Reaction product (RP) indicative of DPP II activity was observed only within lysosome-like organelles. These RP-labelled organelles were only localized in the supra- or para-nuclear regions of the ameloblasts, which corresponds with previous studies using acid phosphatase cytochemistry. However, in contrast with these studies, RP was not detected in the distal region of the ameloblasts, viz., in the Tomes' processes of the secretory ameloblasts or near the ruffled border in the maturation ameloblasts. The transitional ameloblasts were notable for the intensity of staining of their RP-labelled organelles. We propose that DPP II may have a role in programmed cell death which is thought to occur in this transition zone. Biochemical analysis of rat incisor enamel organ homogenates, indicated tissue fixation resulted in an 82% reduction in DPP II activity, although the specific activity of DPP II was not affected.
Anat Rec 1992 Aug
PMID:Cytochemical localization of dipeptidyl peptidase II activity in rat incisor tooth ameloblasts. 162 9

To investigate the functional stages of osteoclasts, the ultrastructural histochemical distribution of the lysosomal enzymes [acid phosphatase (tartrate-sensitive) and neutral phosphatase], the plasma membrane enzymes [alkaline phosphatase, Ca(++)-ATPase, and alkaline ouabain-insensitive p-nitrophenylphosphatase (alkaline p-NPPase)], and the mitochondrial enzyme (cytochrome C oxidase) was evaluated in the chicken tibial metaphysis. Both active-appearing and detached (resting) osteoclasts were studied. Serial sectioning was used to identify detached osteoclasts which were present in the perivascular space. The ultrastructure of detached osteoclasts was similar to that of active osteoclasts, except for the lack of a ruffled border and clear zone, and an altered distribution pattern of small vesicles. Small vesicles were uniformly distributed in the cytoplasm of resting osteoclasts, whereas they were concentrated beneath the ruffled border of active osteoclasts. Alkaline p-NPPase, a marker enzyme for the basal ruffled border, was also apparent on the membrane of small vesicles. However, the vesicles did not possess Ca(++)-ATPase, a marker enzyme for the apical plasma membrane. These findings support the concept that small vesicles serve as a membrane reservoir for the ruffled border membrane. Pre-osteoclasts contained abundant mitochondria and lysosomes, prominent Golgi complexes, moderately developed endoplasmic reticulum, and lacked small vesicles. Pre-osteoclasts appear to fuse with osteoclasts which are attached to the bone surface, but not with detached osteoclasts. The small vesicles, from which the ruffled border arises, are absent from pre-osteoclasts, suggesting that they develop after fusion with pre-existing osteoclasts or after attachment to the bone surface. Alkaline p-NPPase appears to be a marker for differentiation of pre-osteoclasts to mature osteoclasts.
Anat Rec 1991 Nov
PMID:Characterization of the functional stages of osteoclasts by enzyme histochemistry and electron microscopy. 166 72

An ultrastructural, enzymohistochemical, and immunohistochemical study of the ductus epididymis in normal men was undertaken to investigate the characteristics of the apical mitochondria-rich cells (AMRCs). These cells, which differ morphologically from the principal cells (PCs), appear in isolation in the caput epididymidis (5.8 +/- 1.7 cells per cross-sectional duct) and only occasionally in the corpus epididymidis. The morphologic appearance of AMRCs varies from slender cells extending from the basement membrane to the lumen to apical cells without apparent contact with the basement membrane. The former display a round pale nucleus located in the middle of the epithelium; the apical cells have a dark nucleus, which, surrounded by a narrow cytoplasmic band, protrudes into the lumen. The cytoplasm of AMRCs is electron-dense and contains numerous mitochondria surrounded by rough endoplasmic reticulum cisternae. In the apical portion, there are lysosomes, vesicles with an electron-dense granule, and vacuoles showing a variable size and content. The stereocilia are shorter and less numerous than those of the PCs. The AMRCs are similar to the PCs in the intensely positive reaction for the enzymatic activity acid phosphatase, as well as in the lack of reaction for alkaline phosphatase and phosphorylase activities. AMRCs differ from PCs in: (1) a more intense reaction to the enzymatic activities ATPase, NADP, and succinic dehydrogenease, (2) a more intense immunostaining by AE1/AE3 and Ks4.62 anti-cytokeratin antibodies, and anti-estradiol receptor protein (D5) antibodies, and (3) a lower staining affinity for epithelial membrane antigen (EMA) antibodies. No positive immunostaining for the anti-cytokeratin Ks8.6 antibodies was observed in either AMRCs or PCs.
Anat Rec 1991 Sep
PMID:Apical mitochondria-rich cells in the human epididymis: an ultrastructural, enzymohistochemical, and immunohistochemical study. 172 7

Vitamin A-containing lipid droplets in the hepatocytes of rat liver were found to be exocytotically released from the cells in the form of a "lipid droplet--retinol-binding protein (RBP)--immunoreactive complex" following intraportal injection of retinol (17, 33, 67, or 100 micrograms). Evidence that the lipid droplets contain vitamin A was obtained by fluorescence microscopy of vitamin A. Intraportal injection of retinol produced varied numbers and sizes of vacuoles in the hepatocytes. The substance within the vacuoles exhibited a meshwork-like configuration in sections from slices incubated in a medium for revealing acid phosphatase activity or the corresponding control medium and was RBP-immunoreactive and proteinaceous in nature. The occurrence and number of the vacuoles depended on the dosage of injected retinol, being greatest at a dosage of 100 micrograms of retinol and becoming progressively less at dosages of 67, 33, and 17 micrograms. The vacuoles were formed by vacuolization of cisternae of the rough endoplasmic reticulum. The formation of vacuoles reached a maximum 30 min after intraportal injection of 100 micrograms retinol, and the vacuoles and lipid droplets had almost disappeared from the hepatocytes after 90 min. Little or no esterase activity was found in lipid droplets in the hepatocytes before intraportal injection of retinol, but after the injection, lipid droplets that had fused with the vacuoles become strongly positive for this enzyme activity. This suggests that hydrolysis of retinyl esters may occur in the process of complex formation in rat hepatocytes.
Anat Rec 1989 Sep
PMID:The morphology of release of vitamin A-containing lipid droplets by hepatocytes in rat liver. 277 9


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