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Biomedical subjects

J S Rodman

Publications and source records attributed to J S Rodman.

At least 19 recordsLinked to original sources
PubMed1991

Distribution and structure of the vacuolar H+ ATPase in endosomes and lysosomes from LLC-PK1 cells.

Vacuolar proton pumps acidify several intracellular membrane compartments in the endocytic pathway. We have examined the distribution of the vacuolar H+ ATPase in LLC-PK1 cells and the structure of the biosynthetically labeled enzyme in membrane fractions enriched for endosomes or lysosomes. LLC-PK1 cells were allowed to internalize cytochrome c-coated colloidal gold as a marker for endocytic compartments. Proton pumps were identified in these cells by staining the cells with a monoclonal antibody against the vacuolar pump detected with either immunogold or immunoperoxidase techniques. H+ ATPase labeling was seen on structures resembling endosomes and lysosomes, but not on Golgi or plasma membrane. To examine the structure of the H+ ATPase in these compartments, we labeled LLC-PK1 cells for 24 h with [35S]methionine and used a Percoll gradient to obtain fractions enriched for endosomes or lysosomes. H+ ATPase immunoprecipitated from both fractions with monoclonal anti-H+ ATPase antibodies had labeled polypeptides of 70, 56, and 31 kDa. On two-dimensional gels, a comparison of the H+ ATPase from the endosomal and lysosomal fractions revealed that the 70-, 56-, and 31-kDa subunits were similar in both fractions. The results show that the vacuolar H+ ATPase in these cells is distributed primarily in endosomes and lysosomes and that the structure of the enzyme is similar in both compartments.

Animals↗
PubMed1991

Prophylaxis of uric acid stones with alternate day doses of alkaline potassium salts.

Uric acid stone formation ordinarily is prevented by increases in the urinary pH after meals. This postprandial alkaline tide is lost in patients who make such calculi. Single dose, alternate day administration of an alkaline potassium salt will increase urinary pH and simulate this normal physiological mechanism. An important part of the regimen is patient self-monitoring to verify that the urinary pH increases to greater than 6.8, 1 1/2 to 2 hours after the medication is taken. In contrast to multiple dose daily regimens, this mode of base administration is tolerated better and easier to follow. In 17 patients, 7 with the recurrent gravel/colic syndrome and 10 with prior stones, this regimen abolished calculus formation during an average followup of 2 1/2 years. However, further studies are needed before this regimen can be recommended as standard therapy for uric acid stone prophylaxis.

Adult↗
PubMed1990

Immunolocalization of endosomal cathepsin D in rabbit alveolar macrophages.

Intravacuolar proteolysis appears to be an important component of antigen presentation, the activation of peptide hormones, and the conversion of biologically important mediators from inactive precursors. Cathepsin D has been identified in the endosomes of rabbit alveolar macrophages by biochemical analyses [Diment and Stahl, J. Biol. Chem. 260,15311, 1985; Diment et al., J. Biol. Chem. 263,6901, 1988]. Using affinity-purified goat antirabbit cathepsin D IgG, we have localized cathepsin D to the endosomes of rabbit alveolar macrophages. Immunofluorescent staining of frozen sections showed labeling in lysosomes and small vesicles in the periphery of the cell. Label was not seen on the plasma membrane. With immunoperoxidase labeling at the electron microscopic level on cells containing endocytosed mannose-BSA gold, we saw labeling in endosomes and classical lysosomes. When the results were quantitated using immunogold labeling of thin cryosections, we found that the majority of cathepsin D (62.2%) was present in lysosomes, 4.0% in large clear vacuoles, a surprisingly high percentage (29.3%) in small vesicles, 4.9% in endosomes, and none on the plasma membrane. We conclude from this study that, in addition to being present in lysosomes, cathepsin D is present in endosomes and in small peripheral vesicles.

Animals↗
PubMed1989

Urothelial injury from ethylenediaminetetraacetic acid used as an irrigant in the urinary tract.

Although solutions containing disodium ethylenediaminetetraacetic acid (EDTA) will dissolve calcium oxalate stones in vitro, the safety of such solutions as urinary tract irrigants is questionable. These studies were designed to assess the degree of urothelial damage produced by the mildest EDTA formulation which has been reported to be effective. Rabbit bladders were irrigated antegrade via a ureterotomy for 20 hours and then removed for histological examination. A 0.03 M solution of disodium EDTA at pH 7.5 produced considerably more urothelial injury than did an otherwise identical solution of calcium EDTA (p = 0.006). The bladders from the latter group were undistinguishable from those irrigated with saline. As prior saturation of EDTA with calcium completely eliminated the tissue injury, these studies indicate that the same calcium chelating property which makes this chemical effective also makes it toxic. There was enough tissue damage from the relatively mild formulation used to suggest no EDTA solution yet formulated is safe for clinical use.

Animals↗
PubMed1989

The effect of ultraviolet B radiation treatments on calcium excretion and vitamin D metabolites in kidney stone formers.

Several factors could explain the effect of warm, sunny weather on kidney stone formation. To determine the role of sun exposure, we used a light box to administer artificial ultraviolet B radiation during the winter months in New York City. Eleven male stone formers and 7 age- and sex-matched controls received 10 UVB light exposures over a two-week period while maintaining a 400 mg calcium diet. 25-OH vitamin D levels increased significantly (p less than 0.001) in both patients (25.9 +/- 9.8 to 51.6 +/- 14.1 ng/ml) and controls (21.3 +/- 7.1 to 49.6 +/- 3.1 ng/ml). However, 1,25 dihydroxyvitamin D levels rose in the patients (50.8 +/- 14.8 to 55.9 +/- 13.1 pg/ml) but fell (60.1 +/- 6.5 to 49.4 +/- 3.1 pg/ml) in the controls. Only 2 of the 11 patients but all of the controls demonstrated this down regulation of 1,25 dihydroxyvitamin D levels (p less than 0.002). 24,25 dihydroxyvitamin D levels tended to rise in both groups but parathyroid hormone levels were unchanged. There was a trend, which did not reach statistical significance, for parameters of calcium excretion to increase after UVB radiation. 24-hour urinary calcium excretion rose 24% from 140 to 173 mg in the patients and 31% from 113 to 148 mg in the controls. The ratio of calcium/creatinine following a one gram calcium load showed a small increase after UVB radiation from 0.17 to 0.20 in patients and 0.118 to 0.124 in the controls. However, no correlation could be discerned between changes in vitamin D metabolite concentrations and changes in urinary calcium.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1987

Urothelial injury to the rabbit bladder from alkaline irrigants useful in the treatment of uric acid stones.

Irrigation of the urinary tract with alkaline solutions may be used appropriately in some clinical situations to dissolve uric acid stones. Since in vitro studies have suggested that tham-E (pH 10.6) is somewhat more effective than tham (pH 8.6) in promoting dissolution of uric acid, these studies were undertaken to assess the relative toxicities of these two irrigating solutions to the urothelium. Rabbit bladders were irrigated antegrade via a ureterotomy for twenty hours. Tham E produced considerably more mucosal injury than did tham (p = 0.0001). The minimal advantage in rapidity of stone dissolution offered by tham E over tham is more than offset by the considerably increased potential for toxic side effects. The results suggest tham and not tham E should be used for irrigation to dissolve uric acid stones.

Animals↗
PubMed1987

Hypercoagulability produced by treatment with acetohydroxamic acid.

Thrombosis has been observed with unusual frequency in patients receiving acetohydroxamic acid (AHA) for treatment of struvite nephrolithiasis. Because this clinical observation suggests a low-grade state of diffuse intravascular coagulation, standard clinical coagulation assays, platelet counts, and plasma fibrinopeptide A (FPA) levels were measured in patients before and during treatment with AHA. Coagulation assays, fibrinogen, and fibrin-split products were not significantly different when pretreatment and treatment values were compared. FPA levels were significantly elevated and platelet counts were significantly reduced in the treatment period, indicating augmented thrombin activity and consumption of platelets into thrombi or platelet microaggregates. FPA levels were elevated within 24 hours of the initiation of AHA therapy whereas platelet counts did not change significantly within the first 4 days of treatment. The data are consistent with low-grade intravascular coagulation with AHA treatment and suggest that patients treated with this agent should be observed cautiously for signs of thrombosis.

Blood Coagulation↗
PubMed1986

Urothelial injury to the rabbit bladder from various alkaline and acidic solutions used to dissolve kidney stones.

Different irrigating solutions are used clinically to dissolve uric acid, cystine and struvite stones. These studies were undertaken to assess the toxicity to the rabbit bladder epithelium of several commonly used formulations. Test solutions were infused antegrade through a left ureterotomy overnight. Bladders were removed and routine histological sections made. A pH 7.6 solution of NaHCO3 appeared harmless. The same solution with two per cent acetylcysteine produced slight injury. All pH 4 solutions caused significant damage to the urothelium. Hemiacidrin, which contains magnesium, produced less damage than did other pH 4 solutions without that cation. Our data tend to support Suby's conclusions that addition of magnesium reduces urothelial injury even though the presence of magnesium will slow dissolution of struvite.

Acetylcysteine↗
PubMed1986

The membrane composition of coated pits, microvilli, endosomes, and lysosomes is distinctive in the rat kidney proximal tubule cell.

The distribution of a number of membrane proteins on plasmalemmal microdomains (microvilli, coated pits) and in endosomes and lysosomes of the proximal tubule epithelial cell was determined in normal rat kidneys by immunofluorescence and immunoelectron microscopy. Two major brush border proteins, 130 and 94 kD, and gamma-glutamyl transpeptidase were detected on the membranes of the microvilli but were not found on membranes of coated pits. Gp330, the Heymann nephritis antigen, and clathrin were localized in coated pits. The lysosomal membrane glycoprotein, lgp120 (Lewis, V., S. A. Green, M. Marsh, P. Vihko, A. Helenius, and I. Mellman, 1985, J. Cell Biol., 100: 1839-1847) was restricted to lysosomes where it co-localized with beta-glucuronidase. Endosomes, identified by preloading with HRP injected 5-15 min before rats were killed, did not contain detectable amounts of any antigen tested. The distribution of the same proteins was also determined in rats given sodium maleate, which is known to slow or reduce protein absorption by the proximal tubule and to cause vacuolation of the endocytic apparatus. After maleate treatment the distribution of microvillar and lysosomal markers was unchanged, but the coated pit markers were redistributed--gp330 was concentrated in newly formed apical vacuoles, and clathrin was diffusely distributed in the apical cytoplasm or on apical coated vesicles. These findings indicate that the membrane composition of microvilli, coated pits, endosomes, and lysosomes is distinctive in the proximal tubule cell; and that gp330, unlike other known coated pit membrane components, is not transferred to endosomes during endocytosis. After maleate treatment, the coated pits lose their clathrin coats, and the corresponding membrane is internalized.

Animals↗
PubMed1986

Cytoskeletal proteins of the rat kidney proximal tubule brush border.

Cytoskeletal components backing the brush border of the rat kidney proximal tubule cell were identified and compared with those of the well characterized intestinal brush border by immuneoverlay and immunocytochemistry. Antibodies reactive against the intestinal microvillus core components, villin and fimbrin, as well as against the terminal web components, spectrin (fodrin) and myosin, were used. Proteins of similar molecular weight to these intestinal brush border cytoskeletal components were identified in isolated kidney brush borders by immuneoverlay. Spectrin, a major component of the terminal web region of both cell types, was more concentrated in the kidney brush border relative to both actin and myosin. By immunofluorescence, villin and fimbrin were localized in the microvilli, and spectrin and myosin were localized to the terminal web region of the brush border. In addition, spectrin was found along the basolateral membranes of the proximal tubule cell, and myosin was detected in a punctate staining pattern throughout its cytoplasm. By immunoelectron microscopy using immunogold labeling procedures, fimbrin and villin were localized in the terminal web as well as in microvilli, and spectrin and myosin were localized to fibrils in the terminal web. A key difference between the epithelia of the two organs is the extensive network of clathrin coated pits found in the terminal web region of the kidney but not the intestinal brush border. The clathrin-rich terminal web region of the kidney, like the intestinal brush border, proved to be quite stable and resistant to disruption by non-ionic detergents and harsh mechanical treatment.

Animals↗
PubMed1978

Plasma clearance of glycoproteins with terminal mannose and N-acetylglucosamine by liver non-parenchymal cells. Studies with beta-glucuronidase, N-acetyl-beta-D-glucosaminidase, ribonuclease B and agalacto-orosomucoid.

Glycoproteins having mannose and/or N-acetylglucosamine in the terminal non-reducing position [Stockert, Morell & Scheinberg (1976) Biochem. Biophys. Res. Commun. 68, 988--993], and various lysosomal enzymes [Stahl, Schlesinger, Rodman & Doebber (1976) Nature (London) 264, 86--8] are rapidly cleared from plasma by the liver after intravenous administration. A liver cell-separation technique was used to determine the cellular localization of 125I-labelled beta-glucuronidase, ribonuclease B, agalacto-orosomucoid and asialo-orosomucoid. On a specific readioactivity basis, all ligands except 125I-labelled asialo-orosomucoid were enriched in the non-parenchymal cell fraction. Isolated cells, fixed and stained for beta-glucuronidase or N-acetyl-beta-D-glucosaminidase activity after intravenous injection of the enzymes, showed enrichment in the non-parenchymal cell fraction (probably Kupffer cells). After uptake by the non-parenchymal cells, liver lysosomal beta-glucuronidase and N-acetyl-beta-D-glucosaminidase showed degradation half-times of 2.2 and 0.4 days respectively.

Acetylglucosamine↗
PubMed1978

Changes in the kinetics of muscle contraction in vitamin D-depleted rats.

Using an in situ rat soleus neuromuscular preparation, changes in the muscle contraction kinetics in response to vitamine D depletion were studied. For a single isometric contraction, the time-to-peak tension (Tp) and the time-for-recovery-half-way-to-resting tension (T1/2r) were recorded. For a 150 Hz, 300 msec tetanus, the T1/2r was determined. Animals raised on high-calcium, high-phosphate, vitamin D-depleted diets showed prolongation of all parameters. Repletion of vitamin D returned Tp and T1/2r values to normal. Neither dietary calcium deficiency nor thyroparathyroidectomy produced an prolongation of Tp or T1/2r values. Therefore, based upon the experimental data, it appears that vitamin D or one of its metabolites, independent of any effect on the serum calcium or serum phosphate concentration, is necessary for normal muscle relaxation.

Animals↗
PubMed1978

Evidence for receptor-mediated binding of glycoproteins, glycoconjugates, and lysosomal glycosidases by alveolar macrophages.

Alveolar macrophages have been shown to bind glycoproteins and synthetic glycoconjugates (neoglycorpoteins) that have mannose, N-acetylglucosamine, or glucose in the exposed, nonreducing position. Galactose-terminal glycoproteins are not bound. Binding of radiolabeled ligands to cells is nearly completely impaired by the presence of an excess of yeast mannan. Binding is temperature sensitive and proceeds optimally at pH 7.0. Prior treatment of the cells with trypsin severely decreases their capacity to bind ligands. An inhibition assay has been developed, using radioiodinated glucose-albumin conjugate, agalacto-orosomucoid, beta-glucuronidase, and RNase B as ligands. Various glycoproteins have been shown to be effective inhibitors of ligand binding including horseradish peroxidase, agalacto-orosomucoid, beta-glucuronidase, ovalbumin, agalacto-fetuin, and RNase B. RNase A and asialo-fetuin are ineffective as antagonists. The results suggest the presence of a cell surface receptor on alveolar macrophages that binds glycoproteins having terminal sugars with the mannose or glucose configuration.

Animals↗