Mechanism of human erythrocyte hemolysis induced by short-chain phosphatidylcholines and lysophosphatidylcholine

The incorporation and accumulation of a certain amount of short-chain phosphatidylcholine or lysophosphatidylcholine into lipid bilayers of erythrocyte membranes is the first step causing membrane perturbation in the process of hemolysis. Accumulation of dilauroylglycerophosphocholine into membranes makes human erythrocytes "permeable cells"; Ions such as Na+ or K+ can permeate through the membrane, though large molecules such as hemoglobin can not. The "pore" formation was partially reproduced in liposomes prepared from lipids extracted from human erythrocyte membranes; C12:0PC induced the release of glucose from liposomes but did not significantly induce the release of dextran. It was suggested that the phase boundary between dilauroylglycerophosphocholine and the host membrane bilayer or dilauroylglycerophosphocholine rich domain itself behaves as "pores." Erythrocytes could expand to 1.5 times the original cell volume without any appreciable hemolysis when incubated with C12:0PC at 37 degrees C. The capacity of the erythrocytes to expand was temperature dependent. The capacity may play an important role in the resistance of the cells against lysis. The "permeable cell" stage could be hardly observed when erythrocytes were treated with didecanoylglycerophosphocholine and lysophosphatidylcholine. Perturbation induced by accumulation of didecanoylglycerophosphocholine or lysophosphatidylcholine may cause non specific destruction of membranes rather than formation of a kind of "pore."     

Effect of dietary fiber, glucomannan, on absorption of sulfonylurea in man

In order to clarify whether a dietary fiber has any effect upon the intestinal absorption of sulfonylurea, changes in plasma concentration of glibenclamide were determined during a six-hour period in nine healthy volunteers who took 2.5 mg of glibenclamide together with a breakfast and 3.9 g of glucomannan in a form of konjac powder and were compared with those of the control experiment in which the same amount of the hypoglycemic agent was given without the dietary fiber. In the control, mean plasma glibenclamide level increased rapidly, reaching a peak at 60 min and decreased gradually thereafter, whereas an increase in plasma glibenclamide level was blunted in the test experiment, thus plasma concentration of glibenclamide being lower at 30, 60, 90 and 150 min compared with the corresponding value of the control (31.7 +/- 24.5 ng/ml vs 76.4 +/- 25.0 ng/ml at 30 min; 51.3 +/- 35.5 ng/ml vs 120.9 +/- 56.0 ng/ml at 60 min; 60.0 +/- 38.8 ng/ml vs 117.4 +/- 53.1 ng/ml at 90 min; 54.0 +/- 31.5 ng/ml vs 100.7 +/- 46.5 ng/ml at 150 min). Mean plasma glucose concentration was significantly lower at 30 min in the test experiment than in the control despite the lower level of plasma glibenclamide in the former. The results suggest that glucomannan may influence the intestinal absorption of glibenclamide. A dietary fiber must be prescribed in due consideration of these facts.     

13C- and 1H-nmr evidence for all-cis peptide bonds in cyclic tetrapeptide cyclo(L-Pro-Sar)2

Conformations of the cyclic tetrapeptide cyclo(L -Pro-Sar)₂ in solution were studied by ¹H- and ¹³C-nmr spectrometry and model building. The nmr data provide definite evidence that this cyclic peptide exists chiefly in two conformations, namely, a C₂-symmetric conformation and an asymmetric structure. The former was demonstrated to be predominant in polar solvents (100% in Me₂SO-d₆). This structure contains all cis-peptide bond linkages and all trans′ Pro C^α?CO bonds. It represents the first cyclic tetrapeptide in which all four peptide bonds have been found in the cis-conformation. As the polarity of the solvent decreases, the population of C₂-symmetric conformers decreases (88% in CD₃CN and 65% in CDCl₃). At the same time, a minor asymmetric conformer, characterized by cis-cis-cis-trans peptide bond sequences (two cis Sar-Pro bonds, one cis Pro-Sar bond, and one trans Pro-Sar bond), is seen to increase (9% in CD₃CN and 30% in CDCl₃). A proposed predominant conformation in solution for cyclo(L -Pro-Sar)₂ was compared with a crystal structure, as reported in an accompanying paper. Both structures show striking overall similarities.     

Modeling of experiments on biofilm penetration effects in a fluidized bed nitrification reactor

A four-component, diffusion-reaction model with double Michaelis-Menten kinetics was used to describe the experimental data obtained from a laboratory biofilm, fluidized-bed nitrification reactor. Theory and experiment demonstrated that the stoichiometric ratio (3.5 mg O(2)/mg NH(4) (+)-N) can be employed as a criterion to determine whether the limiting substrate is oxygen or ammonia. For the present work, in the range of concentrations where limitation occurred, 4 mg/L NH(4) (+)-N and 14 mg/L O(2), the ratio of oxygen to ammonia in the bulk liquid determined which substrate was penetration-limiting-O(2) if <3.5 and NH(4) (+) if > 3.5. Halforder kinetics with respect to the limiting substrate described the apparent overall rates. Simulations provided biofilm concentration profiles which demonstrated the role of the oxygen-ammonia ratio. Experiments indicated that, generally, high NO(2) (-) concentrations can be expected. These depend on the residence time, biofilm area, and oxygen concentration. This dependency was investigated with the model, as was the parametric sensitivity with respect to the saturation constants. Particularly important for the NO(2) (-) levels were the ratios of the saturation constants for oxygen.     

Studies on the site of 1,25-dihydroxyvitamin D3 synthesis in vivo

Anephric, vitamin D-deficient male rats were injected with a physiologic dose of 25-hydroxy[26,27-3H]vitamin D3 (specific activity of 160 Ci/mmol), and 18-20 h later, intestine, bone, and serum were analyzed by high performance liquid chromatography for 1,25-dihydroxy-[26,27-3H]vitamin D3. Identical studies were carried out using sham-operated rats and rats with ligated ureters. No 1,25-dihydroxy[26,27-3H]vitamin D3 was detected in the tissues from anephric rats, while large amounts were detected in sham-operated and ureteric ligated controls. This result demonstrates that in the nonpregnant rat, 1,25-dihydroxyvitamin D3 is either not synthesized or is synthesized in vanishingly small amounts in bone and intestine in vivo, casting considerable doubt of the physiological importance of reports of in vitro synthesis of 1,25-dihydroxyvitamin D3 by cells in culture derived from bone and elsewhere.     

Transformation-defective Rous sarcoma virus mutants with altered p19 of the gag gene and their inhibitory effect on host cell growth.

Mutants (PH2010, PH2011, PH2012) of Rous sarcoma virus which have a growth-inhibitory effect on chicken embryo fibroblasts were isolated from a temperature-sensitive mutant of the Schmidt-Ruppin strain of Rous sarcoma virus (tsNY68). The growth rate of fibroblasts infected with these viruses was about 50 to 60% of that of uninfected fibroblasts. A morphological difference between mutant-infected and uninfected fibroblasts was observed at logarithmic phase but not at stationary phase. Neither the protein p60src nor its associated protein kinase activity was significantly detected by an immunoprecipitation assay in the cells infected with these mutants. Analysis of the unintegrated DNA of the mutant PH2010 showed that a sequence of about 1.4 kilobase pairs at the src gene region is deleted. Further examination of the viral structural proteins in infected cells as well as in virions by immunoprecipitation and peptide mapping revealed that the molecular size of the Pr76 gag protein of the mutant RSV is smaller than that of the mutant tsNY68 because of partial deletion at the p19 gag gene. The peptide maps suggest that the deleted region of the altered p19 of the mutant is near the carboxy terminal of p19. The amount of Prgp92env synthesized in the mutant-infected cells was about fivefold more than that in tsNY68-infected cells.     

Immunoreactive glucagon in the vascular walls of the rat

We detected immunohistochemically immunoreactive glucagon (IRG) in the smooth muscle of blood vessels and the myoepithelial cell of sweat glands of rats using two antisera against pancreatic glucagon; OAL-123 and 30K. The content of IRG in the blood vessels was found to be 320-1, 270 pg per g wet tissue weight. Filtration of the extracted IRG through a Bio Gel P-30 column yielded a single peak of IRG at 3,500 daltons, the same elution volume of pancreatic glucagon. These findings suggest that the blood vessels of the rats is one of the extrapancreatic sources of IRG in plasma, although physiological role of the IRG is not known.     

Formation of Chlorophyll-Protein Complexes in Greening Cucumber Cotyledons in Light and then in Darkness

The changes in chlorophyll-protein complexes (CPs) in cucumbercotyledons during illumination and subsequent dark incubationwere studied by SDS-polyacrylamide gel electrophoresis. Whenetiolated cucumber seedlings were illuminated, chlorophyll wassynthesized and CPs were formed. In the early phase of greening(6 h of illumination), light-harvesting chlorophyll a/b-proteincomplex (LHCP) was the main GP. As the greening proceeded, P700chlorophyll a-protein complex (CP1) accumulated. When 6-h illuminatedseedlings were transferred to darkness, CP1 accumulated concomitantlywith a decrease in LHCP without new chlorophyll synthesis. Thechanges in the amounts of CPs in the dark became smaller withthe progress of greening and were not observed after 72 h ofillumination. These changes were confirmed by examining thechlorophyll/P700 ratio and the low temperature absorption spectrumof cotyledons. These results suggest that in the early phaseof greening, CPs were unstable and their chlorophyll moleculeseasily exchanged with those of other kinds of CPs. (Received October 14, 1982; Accepted December 1, 1982)