Evidence for different requirements in physical state for the interaction of lipopolysaccharides with the classical and alternative pathways of complement

The influence of the state of aggregation of lipopolysaccharides upon their ability to interact with serum complement via either the classical or alternative pathway was studied. The anticomplement properties of two chromatographically distinct fractions of a phenol-extracted lipopolysaccharide isolated from Serratia marcescens were assessed by means of the standard sheep erythrocyte hemolytic assay and an alternative pathway-selective kinetic assay using rabbit erythrocytes. Both the high molecular weight PI fraction and the lower molecular weight PII fraction exerted anti-complement activity as determined in the sheep erythrocyte assay. Conversion of fractions PI and PII to their more soluble triethylamine salt forms resulted in a decrease in sedimentation coefficients and a corresponding loss of anticomplement activity. Further, the anticomplement activity of fractions PI and PII in the sheep erythrocyte assay was inhibited by polymyxin B, indicating a role for the lipid A region. Unlike the PII fraction, only the PI fraction can activate serum complement via the alternative pathway. This activity is not inhibited by polymyxin B, indicating that the response is not lipid A-mediated. Significantly, solubilization of the PI fraction with triethylamine had no effect on its ability to activate the alternative pathway. These studies clearly demonstrate that the interaction between lipopolysaccharides and serum complement is influenced by the state of lipopolysaccharide aggregation. However, this appears to be the case for lipopolysaccharide activation of the classical pathway but not of the alternative pathway.     

Erythrocytes sedimentation profiles under gravitational field as determined by He-Ne laser. VII. Influence of dextrans, albumin and saline on cellular aggregation and sedimentation rate

The erythrocytes sedimentation profiles (ESP) of normal blood and of blood mixed with saline, albumin (7%), and various molecular weight dextrans of different concentrations, at various height and widths of the sample holder are determined. These observations show that the sedimentation characteristics of the erythrocytes depend on the influence of these substitutes on the plasma and cellular constituents. The normalised aggregation and the sedimentation rate, as determined from these profiles, show that the dextran 40 and dextran 70 retard the erythrocytes sedimentation, for high molecular weight it is similar to that of normal blood and is the maximum for saline. This change for high molecular weight dextrans could be attributed to the enhanced aggregation tendency of erythrocytes and for saline to the enhanced sedimentation due to decrease in the viscosity and density of suspending medium. The influence of the various concentrations of dextrans on these parameters has been determined.     

Osmolality dependence of erythrocyte sedimentation and aggregation in a strong magnetic field

In order to specify the major determinant of the magnetic enhancement of erythrocyte sedimentation observed previously, the dependence of erythrocyte sedimentation rate (ESR) on osmolality was measured under a strong magnetic field. Even at hypotonic osmolality, an increase in ESR due to aggregation was observed in plasma solution as compared with that without aggregation in saline solution. However, the magnetic field did not enhance ESR at hypotonic osmolality, when the cell shape was an isotropic sphere (spherocyte). Thus, we narrowed our search to a mechanism that would explain the enhanced ESR found specifically in anisotropic erythrocytes. It was concluded that the major determinant can only work for anisotropic erythrocytes and is a magnetic field-induced increase in an intermembrane adhesive area due to magnetic orientation of anisotropic erythrocytes.     

Cation specificity of the order—disorder transition in iota carrageenan: effects of kappa carrageenan impurities

The temperature dependence of the optical rotation of carrageenan segments (iota, kappa and iota/kappa mixtures) in different salt solutions (tetramethylammonium or potassium chloride) has been studied. In solutions of pure iota carrageenan (90% iota structure) no evidence of any pronounced cation specificity nor of any thermal hysteresis in the iota carrageenan order-disorder transition was found. However, mixtures of iota and kappa carrageenan displayed a cation specificity similar to that previously observed in iota samples of lesser chemical purity, indicating that kappa impurities in iota samples may be located in regular sequences, rather than randomly in the carrageenan primary structure. Our findings strongly suggest that the order-disorder transitions of iota and kappa sequences in mixed samples occur independently.     

The effect of cholesterol level in erythrocyte membranes on the sedimentation rate, electrophoretic motility, and the rate of potassium ferricyanide reduction

It has been found that with an increase of the molar ratio cholesterol/phospholipids in human erythrocyte membranes in vitro their sedimentation rate in diluted suspensions increases and the reduction rate of potassium ferricyanide decreases, while the electrophoretic mobility of erythrocytes is not changed. The effect of cholesterols discussed in terms of the mechanism of transmembrane transfer of reduction equivalents and non-equilibrium in the double electric layer of erythrocytes surface.     

Effect of potassium leakage on reversible aggregation of human erythrocytes

Changes in the aggregation of human erythrocytes caused by polydextrane were studied under conditions influencing the rate of potassium leakage from cells to medium. It was shown that aggregation decreases as the leakage of potassium ions increases and is completely abolished at leakage rates higher than 2.5-3.0 mmol/l of erythrocytes per hour. The involvement of nonequilibrial electrokinetic phenomena in the inhibition of erythrocyte aggregation by ionic fluxes across erythrocyte surface is discussed. It is proposed that potassium leakage affects the erythrocyte sedimentation rate in clinical investigations.     

Studies on blood components of an air-breathing siluroid fish, Heteropneustes fossilis (Bloch) in relation to body weight.

Different haematological parameters have been studied in relation to the body weight of Heteropneustes fossil (Bloch). The erythrocytes and leucocytes number and haemoglobin concentration increases from lower to higher weight groups. The heart weight also increases along with other blood components. With the unit increase in the body weight of this fish, the heart weight, erythrocytes, leucocytes and haemoglobin increase by a fractional power of 0.85700, 0.13480, 0.13215 and 0.22876, respectively. This shows that haemoglobin increases at a higher rate than erythrocyte number. The coefficient of correlation between body weight and erythrocyte (r = 0.70015), leucocytes (r = 0.95861), haemoglobin (r = 0.96615) and heart weight (0.97577) indicate high degree of correlation. The erythrocytes and leucocytes count and haemoglobin concentration per gram body weight is higher in younger fishes and decrease as the animal grows in size. The haematocrit values and mean corpucsular volumes decrease from lower to higher weight groups, whereas mean corpuscular haemoglobin concentration increases with body weight. The erythrocyte cell surface has a decreasing trend from lower to higher weight group. Due to difference in the rate of decrease of greatest and least diameter of erythrocytes, the elliptical shape of R.B.C. which is common in younger animals, becomes circular in higher weight group. The non-granulocytes increase constantly while the percentage of granulocytes decreases from lower to higher weight groups. The lymphocytes constitute the main bulk of all the leucocytes. The total lymphocytes also increase with the body weight. Spindle cells and monocytes are relatively less in numbers. The percentage of eosinophils, basophils and neutrophils also decrease from lower to higher weight groups.     

Inhibition of carrageenan edema by carrageenan itself]

The influence of two kinin forming agents: iota carrageenan and ellagic acid, on the paw oedema induced by 48/80, an amino-liberator, or by carrageenan iota, has been studied, in the Rat. Ellagic acid and carrageenan, by intraperitoneal injection, reduce the paw oedema induced respectively by 48/80 and carrageenan itself. This inhibition depends on a non-specific "counter-irritation" and not on kininogen stores depletion. Ellagic acid, by intravenous injection, diminishes the oedema induced by carrageenan; swelling due to 48/80, is not affected. So kininogen activation plays some role in the inflammatory processes induced by iota carrageenan. Carrageenan by intravenous injection, suppresses his own inflammatory action but does not influence at all the similar action of 48/80. The anti-inflammatory effect of carrageenan does not exclusively depend on kininogen stores depletion.     

RNA synthesis in maturing avian erythrocyte nuclei

The rate of RNA synthesis and its inhibition by α-amanitin in the nuclei of mature and immature avian erythrocytes are increased with the increase in ionic strength of incubation medium. Polyacrylamide gel electrophoresis indicates that heterogeneous species of RNAs are synthesised in the mature and immature erythrocyte nuclei. However, a large number of high molecular weight RNAs are synthesised in the nuclei of immature erythrocytes. Elution profiles on poly(U)-sepharose chromatography indicate that the RNAs synthesised in the nuclei of two types of cell contain poly(A) segments. Sixteen per cent of mature erythrocyte nuclear RNA syntbesised are polyadenylated, while it is 13% in immature erythrocyte nuclei. However, the total RNA synthesised is 2–3 fold higher in immature erythrocyte nuclei than that in mature erythrocyte nuclei.     

The Sedimentation Pattern of D-Glucose-6-Phosphate Dehydrogenase from Bovine Fetal and Adult Erythrocytes

Erythrocytes from bovine fetuses contain about 2.4 times higher D-glucose-6-phosphate dehydrogenase activities than erythrocytes from adult cows and bulls. Studying whether this is due to the existence of a special fetal type of enzyme or an increased amount of enzyme in fetal erythrocytes, the sedimentation coefficients of the enzymes have been estimated by s-zonal ultracentrifugation, and compared to normal and deficient human erythrocyte D-glucose-6-phosphate dehydrogenase, s-zonal ultracentrifugations have been performed with a computer optimized isokinetic sucrose gradient. The mainlines in the program used for calculation of sedimentation coefficients are described. Bovine fetal and adult erythrocyte D-glucose-6-phosphate dehydrogenase was found to have the same sedimentation coefficient of 7.4 S which is different from the sedimentation coefficient of 6.4 S of both human types of the enzyme. The sedimentation coefficients of 6-phospho-D-gluconate dehydrogenase from bovine fetal, bovine adult and human erythrocytes were 6 S for all three types of this enzyme. By cellulose acetate electrophoresis bovine fetal and adult D-glucose-6-phosphate dehydrogenase show the same mobility, again differing from the normal and deficient human type. The results of these experiments show that bovine fetal and adult erythrocytic D-glucose-6-phosphate dehydrogenase with respects to molecular parameters are closely related and perhaps identical enzymes.     

Effects of Aggregation on Blood Sedimentation and Conductivity

The erythrocyte sedimentation rate (ESR) test has been used for over a century. The Westergren method is routinely used in a variety of clinics. However, the mechanism of erythrocyte sedimentation remains unclear, and the 60 min required for the test seems excessive. We investigated the effects of cell aggregation during blood sedimentation and electrical conductivity at different hematocrits. A sample of blood was drop cast into a small chamber with two planar electrodes placed on the bottom. The measured blood conductivity increased slightly during the first minute and decreased thereafter. We explored various methods of enhancing or retarding the erythrocyte aggregation. Using experimental measurements and theoretical calculations, we show that the initial increase in blood conductivity was indeed caused by aggregation, while the subsequent decrease in conductivity resulted from the deposition of erythrocytes. We present a method for calculating blood conductivity based on effective medium theory. Erythrocytes are modeled as conducting spheroids surrounded by a thin insulating membrane. A digital camera was used to investigate the erythrocyte sedimentation behavior and the distribution of the cell volume fraction in a capillary tube. Experimental observations and theoretical estimations of the settling velocity are provided. We experimentally demonstrate that the disaggregated cells settle much slower than the aggregated cells. We show that our method of measuring the electrical conductivity credibly reflected the ESR. The method was very sensitive to the initial stage of aggregation and sedimentation, while the sedimentation curve for the Westergren ESR test has a very mild slope in the initial time. We tested our method for rapid estimation of the Westergren ESR. We show a correlation between our method of measuring changes in blood conductivity and standard Westergren ESR method. In the future, our method could be examined as a potential means of accelerating ESR tests in clinical practice.     

Study of erythrocyte sedimentation behavior by piezoelectric crystal impedance sensor

Based on the impedance characteristic of erythrocytes at high frequency, the response of piezoelectric crystal impedance (PCI) sensor in the erythrocyte suspension was derived and verified experimentally. A method of using PCI sensor to investigate erythrocyte aggregation-sedimentation phenomenon was proposed. From the frequency response of the PCI sensor, the erythrocyte aggregation time and sedimentation rate could be obtained during erythrocyte aggregation and sedimentation. With the present method, the effects of the erythrocyte deformability, the osmotic pressure and the coexisting macromolecules on the erythrocyte sedimentation rate were studied. The results show that the PCI sensor possesses some advantages, such as good sensitivity, simplicity of use and no thermal effect for the impedance study of erythrocyte aggregation and sedimentation.     

Physicochemical and functional properties of hemoglobin of mice infected with Ehrlich's carcinoma]

Injection of white mice with Ehrlich's carcinoma triggers an increase in the mice blood erythrocyte fraction suspended in the 14% sucrose concentration zone. As established by acid erythrograms, a quantitative increase of this red blood cell population is due to an increased rate of erythroblast maturation and occurrence of immature cell forms in the blood stream. As shown by alkali denaturation of hemoglobin, tumour development causes an increase in alkali-resistant hemoglobin fraction in the erythrocytes. On the basis of the data on alkali denaturation of hemoglobin, it is suggested that in the infected mice increased rate of erythrocyte maturation go in line with selective binding of alkali resistant hemoglobin fraction to red cell membrane.     

Blood dyscrasia in a teleost, Colisa fasdatus after acute exposure to sublethal concentrations of lead

Colisa fasciatus , a freshwater teleost, were exposed for 90-h to 15 mg l⁻¹ (0.79 of the96-h LC₅₀ value) lead nitrate under static test conditions. The treatment resulted in decreased ( P≤ 0.001) erythrocyte counts, haematocrits and haemoglobin contents of the moribund fish. Toxicity of the metal was also characterized by significant ( P <0.001) acceleration in erythrocyte sedimentation rate, numerical increase in the number of immature erythrocytes in circulation, lysis and degeneration of erythrocytes, and an increase in the hepatosomatic index. Leucocyte and thrombocyte counts, the number of lymphocytes, and blood clotting times were not significantly different between experimental and control fish. Haemolytic anaemia and an overt increase of circulating immature erythrocytes can be used in monitoring lead poisoning in fish.     

A new multimeric hemagglutinin from the coelomic fluid of the sea urchin Anthocidaris crassispina

A hemagglutinin was purified from the coelomic fluid of the sea urchin Anthocidaris crassispina by ion-exchange chromatography on DEAE-cellulose and affinity adsorption to glutaraldehyde-fixed ghosts of human erythrocytes, followed by elution with 10 mM EDTA. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, it showed a single protein band with a molecular weight of 13 000 and 26 000 in the presence and absence of 2-mercaptoethanol, respectively. The molecular weight of the native protein with a hemagglutinating activity was determined to be 300 000 by sedimentation equilibrium analysis. Its sedimentation coefficient, S0(20),w, and Stokes radius were 13.7 S and 5.5 nm, respectively. The hemagglutinating activity of this protein required calcium ions. When calcium ions were depleted, no activity was observed and its sedimentation coefficient, S0(20),w, decreased to 11.4 S while its Stokes radius increased to 6.7 nm without a change in its molecular weight. The purified hemagglutinin agglutinated human erythrocytes regardless of their ABO and MN blood types. The hemagglutination reaction was not affected appreciably by various simple sugars but was inhibited by tryptic fragments released from human erythrocyte membranes. The results of alkaline borohydride treatment of the inhibitory tryptic fragments showed that the receptor sites for this hemagglutinin were mainly composed of alkali-labile carbohydrate chains with the structure AcNeu alpha 2----3Gal beta 1----3(AcNeu alpha 2----6)GalNAc----serine (or threonine).(ABSTRACT TRUNCATED AT 250 WORDS)     

Hypotensive and edematogenic effects of carrageenan in the irradiated rat]

Iota carrageenan induces the same hypotensive effect in leucopenic and thrombopenic rats as in normal animals. This hypotensive activity depends on kininogen-kinins activation. In the irradiated rats, the inflammatory response to iota carrageenan is reduced. The blood cells are thus responsible of the major part of the late inflammatory effect. There is non parallelism between this hypotensive and the oedematogen activities of iota carrageenan in the rat.     

The mechanism of blood cell sedimentation. XVIII. Sedimentation effect of fibrinogen plasminolysis products]

Various high molecular weight plasmin degradation products of fibrinogen were isolated by chromatographic procedures and investigated to what extent they influence the sedimentation rate of erythrocytes in heparinized serum. Among the early plasminolysis products Fragment X accelerated the sedimentation rate considerably although it was less effective than fibrinogen. Fragment Y showed a weaker but clearly demonstrable agglomerine activity. The late plasminolysis products D and E were ineffective, if applied alone and in concentrations equivalent to the fibrinogen content of plasma. However, they promoted the sedimentation effect of fibrinogen. If present in considerably higher concentrations are Fragments D and E, in absence of fibrinogen, also accelerated the sedimentation rate of erythrocytes significantly. A mixture of D and E was not more effective than the single compounds.     

Alteration of Cl- transport in erythrocytes from patients with Huntington's disease

Rate constants for Cl- transport across erythrocyte membranes were measured in experiments performed simultaneously on normal erythrocytes (control) and erythrocytes from patients with Huntington's disease (HD). The rate constants were observed to decrease in HD cells during aging in vitro from 12% higher to lower than normal values. Young HD cells, separated from blood, quickly decreased their permeability for anions by about 30%, while old cells seemed to increase it. Rate constants in control erythrocytes remained steady and equal in both fractions. A hypothesis has been proposed that there might be two populations of erythrocytes in HD blood, one abnormal, characterized by an initially high anion permeability, which decreases significantly during the cell life. A theoretical analysis of the results has suggested that the abnormal fraction of erythrocytes in HD blood may be associated with an increased population of stomatocytes observed by electron microscopy.     

Chemical and physical characteristics of carrageenan extracted from Eucheuma spinosum harvested from three different Indonesian coastal sea regions

Carrageenan extracted from Eucheuma spinosum harvested from three different coastal sea regions, where this alga has been mainly cultivated, were determined for their chemical and physical characteristics. The carrageenan was extracted from the seaweed using hot alkali followed by precipitation, drying, and milling. The carrageenan properties were determined in terms of yield, ash, mineral, sulfate content, functional group, molecular weight, and viscosity profile. Physical characteristics of carrageenan were evaluated by a texture analyzer for gel strength and a rapid visco analyzer for viscosity. The yield of carrageenan from Sumenep (34.81 ± 5.83%) and Takalar (37.16 ± 3.26%) was found to be relatively higher than that of Nusa Penida (25.81 ± 1.93%). The calcium content was higher than magnesium, potassium and sodium content, and no cadmium, lead, mercury, and arsenic detected in all carrageenan. The ash content was around 29%; while, the sulfate content was in the range of 30–32%, and those were not different in all carrageenan. The presence of sulfate content was identified by FTIR at absorption band of 1373 cm^?1. It was found that the molecular weight of carrageenan from Takalar were relatively higher and the gel strength of carrageenan from Takalar were significantly higher than that of carrageenan from Nusa Penida and Sumenep. Likewise, upon cooling from 80 to 20°C, the viscosity profile of carrageenan from Takalar characterized by higher viscosity compared to that of carrageenan from Sumenep and Nusa Penida. These results indicated that carrageenan from Nusa Penida, Sumenep, and Takalar were identified as iota‐carrageenan with similar physico‐chemical characteristics except for the gel strength, viscosity profile upon cooling from 80 to 20°C and the yield.