Leucine aminopeptidase activity is diminished in aged hog, beef, and human lens.
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Biomedical subjects
Publications and source records attributed to A Taylor.
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The crystallization of leucine aminopeptidase from hog kidney is reported for the first time. The crystals which diffract to 4-A resolution have the space group P2(1)2(1)2(1) (a = 186.3 A, b = 223.2 A, and c = 80.5 A) and contain four hexamers per unit cell, or one per asymmetric unit. Electron micrographic images of hog kidney leucine aminopeptidase are indistinguishable from micrographs of beef leucine aminopeptidase taken under the same conditions (10). These reveal an equilateral triangle of about 85 A per side, seemingly made of three 40-A diameter spheres. This triangle is circumscribed by another concentric, less-dense triangle of 120 A per side which is rotated 60 degrees with respect to the inner triangle. Immunodiffusion and microcomplement fixation assays indicate that the two enzymes share greater than 90% amino acid sequence homology. This similarity is corroborated by peptide maps of tryptic fragments of the radioiodinated enzymes. The model of the quaternary structure proposed to explain the appearance of electron micrographs of single molecule and crystalline bovine lens enzyme also describes the hog kidney enzyme equally well. That the model of leucine aminopeptidase originally proposed for the beef enzyme also can be used to describe hog kidney leucine aminopeptidase crystal packing in the highly anisometric unit cell provides further corroboration that leucine aminopeptidase in these two species is a hexamer based on two trimers each made of three bilobal promoters.
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The objective of this study was to examine anatomically the distribution of afferent neurones in the mesencephalic nucleus of the fifth nerve (Mes V). HRP was applied, in separate experiments, to the inferior alveolar, infraorbital, and masseter nerves, and injected into the masseter muscle and periodontal ligament. Following application of HRP to the masseter muscle and masseter nerve, labelled cells were found in the ipsilateral motor nucleus of the fifth nerve and in the ipsilateral Mes V. Labelled cells in Mes V, identified as belonging to proprioceptor afferents from jaw-closing muscles, were distributed throughout the full extent of the nucleus. Following application of HRP to the inferior alveolar nerve, infraorbital nerve, and periodontal ligament, labelled cells were found in the ipsilateral trigeminal ganglion and Mes V, and the latter identified as belonging to periodontal receptor afferents. In contrast to the distribution of spindle afferent somata, they were restricted to the caudal region of Mes V. The differential distribution of afferent neurones within Mes V demonstrated in this study confirms previous electrophysiological findings, and its significance is considered.
Gelatin coupled to Sepharose has been used to isolate [35S]methionine-labeled polypeptides of Mr = 47,000, 56,000, 62,000, and 65,000 from the 12,000 X g supernatant of detergent extracts of mouse embryo parietal endoderm cells. The polypeptides can also be recovered from various established cell lines which synthesize type IV procollagen, and in these cells the Mr = 47,000 polypeptide is the major gelatin-binding component. Several lines of evidence, including the results of continuous labeling and pulse-chase experiments, show that the polypeptides are not derived by proteolytic cleavage of larger precursors and are distinct from fragments of fibronectin. The Mr = 47,000, 62,000, and 65,000 polypeptides all contain N-linked oligosaccharide side chains, as judged by their labeling with [3H]mannose and their sensitivity to tunicamycin. The Mr = 62,000 and 65,000 polypeptides can be resolved by two-dimensional gel electrophoresis into species with different isoelectric points, and the Mr = 47,000 has a pI of 7.5-8.0. None of the gelatin-binding polypeptides appear to accumulate in the culture medium, and the Mr = 47,000, 62,000, and 65,000 species are labeled in a lactoperoxidase-catalyzed iodination of intact cells, suggesting that they are associated with the cell surface. Only the Mr = 47,000 glycoprotein binds to native type IV collagen. Possible functions for these surface components in vivo are discussed.
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This study was designed to assess the relative influence of meal weight and caloric content on gastric emptying of liquid and solid meals in man. A dual radioisotopic method which permits noninvasive and simultaneous measurement of liquid- and solid-phase emptying by external gamma camera techniques was employed. Nine healthy volunteer subjects ingested 50-, 300-, and 900-g lettuce and water meals adjusted to either 68, 208, or 633 kcal with added salad oil. The following observations were made: (1) absolute emptying rates (grams of solid food emptied from the stomach per minute) increased directly and significantly with meal weight; (2) increasing meal total caloric content significantly slowed solid food gastric emptying but did not overcome the enhancing effect of meal weight; and (3) liquid emptying rates were uninfluenced by meal total kcal amount.
Antisera specific for leucine aminopeptidase (LAP) were prepared. Using these antisera in immunodiffusion tests, the identity of LAP isolated from beef lens and kidney is demonstrated. The same pertains to hog lens and kidney LAP. As indicated by only partial fusion of the immunoprecipitates in immunodiffusion plates, leucine aminopeptidases isolated from hog and beef are antigenically similar but not identical. These tests also indicate substantial similarity between a component in human lens homogenates and bovine or hog LAP. Microcomplement fixation tests corroborate these observations and indicate that, under these conditions, LAP from beef lens and kidney, or hog lens and kidney, are indistinguishable. However, there is an approximate 8.5% amino acid sequence difference between beef and hog LAP. Microcomplement fixation tests with human lens homogenate also corroborate immunodiffusion results and indicate an approximate 19% amino acid sequence difference between beef and human LAP. These data establish that LAP is a species-specific enzyme and they indicate that it is not organ specific. Maximal complement fixation occurs at approximately 0.1 microgram antigen per tube in those assays in which pure aminopeptidases were tested. This permits standardization of the microcomplement fixation assay for LAP under these conditions. Maximal complement fixation occurs at 160-200 micrograms human lens homogenate per tube. Assuming that in this quantity of homogenate there is 0.1 microgram LAP, then it can be calculated that LAP comprises about 0.05% of the lens protein. This agrees closely with the percentage of LAP in hog and beef lenses. Thus, the reduced LAP activity reported for human lens tissue appears not to result from an absence of the enzyme but rather, may be due to diminished catalytic competency of the enzyme in aged human lens tissue (see Taylor and Juhngen , 1984). The unit evolutionary period, 4.7-5.8 X 10(6) years, indicates that LAP has been highly conserved during evolution.
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We determined that mouse lymphoid cell lines can be transfected at high efficiencies (10-70%) by a polyoma virus shuttle vector. With this vector, we obtained expression of a cloned mouse alpha heavy chain gene transfected into cell lines representative of all stages in B-lymphocyte development, a T-cell lymphoma line, and 3T3 fibroblasts. Heavy chain gene expression in transfected light chain-producing myeloma cells occurred at levels comparable to those in IgA-secreting myeloma cells. Heavy chains produced in transfected myeloma cells were associated with light chains in membrane-bound IgA. While T-lymphoma cells and fibroblasts were transfected at similar efficiencies to B cells, significantly lower levels of alpha heavy chains were produced. This immunoglobulin gene transfection system provides a powerful approach for defining important regulatory regions in immunoglobulin genes and for identifying lymphoid cell factors involved in immunoglobulin gene expression in B-lymphocyte development.
A 29-year-old gardener developed acute renal failure following the ingestion of 'Mersil', a combination of mercurous and mercuric chloride, achieving a plasma mercury concentration of 22,000 nmol/litre (400 micrograms/litre). Haemodialysis and charcoal haemoperfusion were ineffective in removing mercury despite prior treatment with the chelating agent dimercaprol. The acute renal failure resolved after 10 days and there are no residual sequelae.
Polyporic acid, atromentin, bovinone, and oosporein are common metabolic products of a number of species of fungi. The related compound cochliodinol and its congeners are produced by several Chaetomium spp. These quinonoid metabolites have been shown to inhibit the growth and metabolism of a range of bacterial genera. The antibiotic activity of the quinones depends on the substituents at the 3 and 6 positions of the 2,5-dihydroxy-1,4-benzoquinone ring; in aerobic systems the activity appears to be inversely proportional to the polarity of the metabolite. It has been shown that reduction of the quinone to the hydroquinone changes the antibiotic activity of these metabolites but does not abolish it. Contrary to previous reports, the activity of these hydroquinones is not reversed by cysteine.
Diuretic radionuclide renography is accurate in the assessment of nephroureteral dilatation. We have extended this technique to include evaluation of nephroureteral dilatation noted on the bone scans of two patients.
Electrical stimulation of low threshold, fast afferents in the inferior alveolar nerve produce very short latency synaptic field potentials in the cerebellar cortex anterior lobe typical of mossy fibre inputs. The afferents concerned are from periodontal receptors, some of which project directly without relay. Muscle afferents do not show this direct projection. The functional significance of this arrangement is discussed, bearing in mind the parallel with the direct projection of some first order vestibular afferents. It is suggested that the specialisation seen may be the basis for sensory calibration of spindle inputs by means of periodontal afferents.
During sustained coronary occlusion in canine preparations, the extent of regions that fail to show contrast enhancement when imaged by supra-aortic hydrogen peroxide contrast echocardiography (SHPCE) has been shown to correlate well for single cross sections with the extent of malperfused myocardium "at risk" of infarction. In the present study, SHPCE was investigated as a means of determining the fraction of total left ventricular mass at risk during occlusion. Since necrotic tissue has low blood flow even when reperfused, we also investigated the potential of quantitating the extent of infarcted myocardium by measuring the extent of contrast defects seen with SHPCE performed during reperfusion. In 20 dogs the fraction of the left ventricle showing a contrast defect during coronary occlusion correlated well with the fraction of the left ventricular mass "at risk" by an autoradiographic technique (autoradiography = 0.83 echocardiography + 8.6%; r = .89, SEE = 4.5%). SHPCE was also performed after 3 hr of reperfusion following occlusions varying in duration from 60 to 150 min. The fraction of the ventricle showing a contrast defect during reperfusion predicted the infarcted portion of the left ventricle as shown by triphenyl tetrazolium chloride staining (% left ventricle infarcted = 0.81 echocardiography + 3.3%; r = .84, SEE = 5.3%). Observer variability for the fraction of the ventricle showing a contrast defect was excellent during both occlusion and reperfusion. The ratio of the left ventricular extent of contrast-negative regions during reperfusion and occlusion was used to calculate a necrosis-to-risk index in vivo that correlated relatively well with the myocardial necrosis-to-risk ratio determined morphologically (r = .77, SEE = 16%).(ABSTRACT TRUNCATED AT 250 WORDS)
There is increasing interest in initial therapy of hypertension with sympatholytic agents and the influence of antihypertensive therapy on cardiac and renal function. We treated 26 men with essential hypertension with either guanabenz alone (n = 14) or propranolol alone (n = 12) and assessed blood pressure and renal perfusion before and after 5-7 weeks of treatment. Cardiac performance was evaluated for the guanabenz-treated patients. Both drugs substantially reduced blood pressure without weight gain. During guanabenz therapy, glomerular filtration rate and renal blood flow were preserved, with a fall in renal vascular resistance (from 12,100 +/- 1,500 to 9,300 +/- 1,190 dyne X s X cm-5, p less than 0.01). Propranolol decreased glomerular filtration rate (from 95 +/- 11 to 70 +/- 6 ml/min, p less than 0.05) without significant change in renal blood flow or renal vascular resistance. In guanabenz-treated patients, there was a decline in left ventricular mass (from 290 +/- 23 to 257 +/- 14 g, p = 0.067). Thus, both agents are effective initial therapy in hypertension. Guanabenz treatment also was associated with reduced renal vascular resistance and left ventricular mass.