Effect of temperature on the metabolism of proteoglycans in explants of bovine articular cartilage

The turnover of proteoglycans in bovine articular cartilage was determined in explant cultures, maintained at 32 degrees C or 37 degrees C. Both the rate of proteoglycan synthesis and the release of newly synthesized proteoglycans were decreased in cultures incubated at 32 degrees C compared to 37 degrees C. At both temperatures the newly synthesized proteoglycans were similar in hydrodynamic size and chain length of the glycosaminoglycans. However, the ratio of 6-sulfated disaccharides over 4-sulfated disaccharides of the newly synthesized glycosaminoglycans, differed less from the endogenous ratio at 32 degrees C than at 37 degrees C. At both temperatures, the incorporated 35S-sulfate is released from explants in two pools. Twenty-three percent of the 35S-radiolabel was released into the culture medium during an initial short phase (t1/2 = 1.1 day at 32 degrees C, 1.3 day at 37 degrees C), 77% had a much longer half-life. The lowered temperature markedly decreased the release of 35S-sulfate with a slow turnover (t1/2 = 60 days at 32 degrees C, 38 days at 37 degrees C).     

Thermal Regulation of Spontaneous Spike Frequency and Concomitant Changes in the Amplitude of Spikes from Cortical Neurons

Studies using living slices of guinea pig sensorimotor cortex showed that changes in temperature from 24°C to 37°C produced stepwise increases in neuron spike frequency at two temperature zones: 27–29°C and 34–36°C. Changes in spontaneous activity were accompanied by decreases in spike amplitude at t < 27°C and t > 34°C. After cooling to 24°C, spike amplitude generally recovered completely when the temperature was increased to 32–34°C. The decrease in spike amplitude at t > 35°C could not be restored by decreasing the temperature. It is suggested that these effects are associated with the K⁺ permeability of neuron membranes.     

Hyperglycemia Reduces Proteoglycan Levels in Tendons

Rationale: Diabetic tendinopathy is characterized by increased stiffness, thickness, and excess calcification of affected tendons. We investigated the hypothesis that proteoglycans (PGs), as key regulators of tendon structure and calcification, are altered in diabetic tendons.

Methods: Adult porcine patellar tendons were incubated in iso-osmolar media with high or normal glucose levels for 2 weeks. The PG fraction was isolated and analyzed. Protein and mRNA levels of five PGs were measured. PG production was assessed in primary tenocyte monolayers by ³⁵S-sulfate labeling in high and normal glucose conditions with and without exposure to advanced glycation end-products (AGEs). Levels of transforming growth factor β, which commonly mediates some effects of hyperglycemia, were also measured and the effects of free radical scavengers on ³⁵S-sulfate incorporation were determined.

Results: PG levels were significantly decreased in tendons exposed to high glucose media compared with tendons in iso-osmolar control media. Relative quantities of individual PGs were unchanged by exposure to hyperglycemia and mRNAs for PGs were variably affected. High glucose media decreased PG production by tenocytes as measured by ³⁵S-sulfate labeling, whereas AGE-modified type I collagen and free radical scavengers had no effects. Hyperglycemic conditions increased levels of transforming growth factor β1 in an AGE-independent manner.

Conclusions: Hyperglycemia produces a reduction in PG levels related to decreased synthesis or sulfation of glycosaminoglycans, which may contribute to the tendon pathology observed clinically in diabetes.     

Thermokinetic profile of the yeast Rhodotorula ferulica and its modification by tributyltin-oxide (TBTO)

The temperature range of growth of Rhodotorula ferulica on vanillic acid shrank from 5.3–31.6 °C to 11.1–26.0 °C in the presence of TBTO (tributyltin-oxide) at 23.4 μM (the maximum tolerated by the organism). However, it took just 5 μM of the drug to transform the thermokinetic profile from associative (cardinal temperatures: t_op = 27–28°C, t_{min d} = 29.5°C, t_{max f} = 32.5 °C, t_{max i} = 35 °C) into dissociative (t_{min d} = 34 °C, t_max = 30 °C), the specific growth rates decreasing by one order of magnitude. In the supraoptimal temperature range (when exponential growth concurred with thermal death) the yield coefficients decreased by 70% on vanillic acid, rather than on glucose. The results are consistent with the hypothesis that the target for TBTO in this yeast is different from the thermal death target.     

Sulfation and transport of basement membrane proteoglycans,as visualized by 35S-sulfate radioautography in the endodermal cells of the rat parietal yolk sac

In the hope of clarifying the biogenesis of basement membrane proteoglycans, an injection of ³⁵S-sulfate was given to concepti of 12-day gestant Sherman rats. The parietal wall of the yolk sac (including endodermal cells and the associated basement membrane known as Reichert's membrane) was removed at times varying from 7 min to 24 hr after injection and processed for electron microscopic radioautography. Silver grains were counted over the organelles of endodermal cells as well as over Reichert's membrane. Between 7 min and 2 hr after ³⁵S-sulfate injection, radioactivity was observed in the endodermal cells, while from 4 to 24 hr it was mostly present in Reichert's membrane. Detailed distribution of the cellular radioactivity at 7 and 15 min showed about 20% in the rough endoplasmic reticulum (rER), 60% in the Golgi apparatus, and 8% in secretory granules. The radiactivity present in rER and Golgi apparatus decreased to low levels by 2--4 hr after injection. In secretory granules, radioactivity increased to reach a peak at 2 hr and then declined; moreover, only the granules associated with the trans Golgi face were radioactive at early time intervals, while those scattered through the cytoplasm and along the cell surface became radioactive at later times. Between 4 and 24 hr, radioactivity became negligible over all cell organelles, while it was collected in Reichert's membrane. Biochemical reports indicate that when ³⁵S-sulfate is added to organ cultures of Reichert's membrane and endodermal cells, about 90% of the incorporated which these proteoglycans acquire sulfate are likely to be those labeled at 7 min after ³⁵S-sulfate injection, that is, the Golgi apparatus and to a lesser extent the rER, whereas some labeling of the secretory granules located at the trans Golgi face is explained by rapid acquisition of sulfated proteoglycans from the Golgi apparatus. Label later appears in the secretory granules along the cell surface and, eventually, in Reichert's membrane. It is concluded that secretory granules transport newly formed proteoglycans from the Golgi apparatus to the outside for deposition into Reichert's membrane.     

A new application of Gompertz function in photohemolysis: the effect of temperature on red blood cell hemolysis photosensitized by protoporphyrin IX

Photosensitization by protoporphyrin IX (PpIX) is accelerated at different irradiation temperatures, different dark incubation temperatures (T _inc) and different irradiation times. The applicability of Gompertz function to the fractional photohemolysis ratio, a and the rate of fractional photohemolysis, b is found to be the most appropriate model to fit the experimental data with minimum parameters and minimum errors. The reduction in Gompertz parameters, the fractional ratio values of a, and increase in the fractional rate values b, for 20 μM PpIX irradiated with black light at low irradiation temperature 5°C and higher T _inc 37°C was noticed. The parameter a has higher values at lower irradiation time and lower irradiation temperatures which indicates a longer photohemolysis process and longer t ₅₀. Values of the parameter b were found to be strongly temperature-dependent, and always increase with increasing irradiation time and T _inc with lower values at lower irradiation time and lower T _inc. There are no significant changes in the lysis of RBCs process at irradiation temperatures equal to or higher than 35°C. Similarly, no significant change on t ₅₀ at higher irradiation time at T _inc 24 and 37°C. In conclusion, Gompertz analysis technique adapts to study the photohemolysis process at different conditions as a best-fit model.     

Polymerization of 1,3‐Pentadiene Initiated by Aluminium Trichloride in Nonpolar Solvent: Pseudo‐Control is Explained by Continuous Grafting

Summary: The cationic polymerization of 1,3‐pentadiene initiated by AlCl₃ was studied in nonpolar solvent. It was previously shown that at room temperature the active species were long‐lived and that the number‐average molar mass of the polymer chains was increasing with the polymerization yield. In order to explain this apparent control, the macromolecules were labeled with a transfer agent, triphenylamine (NPh₃). The latter binds to active species by electrophilic aromatic substitution. The labeling of the polymer chains indicated that at 20 °C the polymer chains mainly contained one NPh₃ molecule per macromolecule while the NPh₃ content was higher for the high molar mass chains due to a “grafting from” polymer transfer mechanism. Thus, the pseudo‐control was assigned to the branching reactions. The labeling process by NPh₃ also succeeded at ?10 °C. Whereas at ?10 °C a dialkylation of NPh₃ was observed, a trialkylation at 20 °C was obtained. The analysis of the polymer microstructure at both temperatures highlighted an interaction between the active centers and NPh₃. This paper also describes a process to synthesize tri‐arm stars polymers by cationic polymerization.

RI SEC chromatograms of soluble polymers synthesized at 20 °C in the presence of NPh₃ with increasing reaction times (r = [NPh₃]/[AlCl₃] = 1); (a) t = 0.25 h, (b) t = 0.5 h, (c) t = 1 h, (d) t = 2 h, (e) t = 18 h, (f) t = 48 h; [AlCl₃] = 2.3 × 10^?2 mol · L^?1, [1,3‐pentadiene] = 1.6 mol · L^?1, pentane.     

Comparative NMR studies of diffusional water permeability of red blood cells from different species. X. Camel ( Camelus dromedarius ) and alpaca ( Lama pacos )

The diffusional water permeability (P _d) of camel and alpaca red blood cells (RBCs) was measured by a doping nuclear magnetic resonance (NMR) technique on control cells and following inhibition withp-chloromercuribenzene sulphonate (PCMBS). The values ofP _d were, in the case of alpaca RBC≈4.6×10⁻³ cm/s at 25°C, 5.4×10⁻³ cm/s at 30°C, 6.6×10⁻³ cm/s at 37°C and 7.7×10⁻³ cm/s at 42°C. In case of camel RBC the values ofP _d where ≈4.2×10⁻³ cm/s and 9.0×10⁻³ cm/s at 42°C. Systematic studies on the effects of PCMBS on water diffusion in camel RBC indicated that the maximal inhibition was reached in 45 min with 1–2 mm PCMBS. The values of maximal inhibition were around 47% at 25°C and 68% at 30°C for alpaca RBC and around 62% at 25°C and 56% at 37°C for camel RBC. The basal permeability to water of alpaca RBC was estimated at around 2.6×10⁻³ cm/s at 25°C, 1.7×10⁻³ cm/s at 30°C and of camel RBC as 1.8×10⁻³ cm/s at 25°C and 3.0×10⁻³ cm/s at 37°C. The values of the activation energy of water diffusion (E _{a, d}) were around 23 kJ/mol for camel and 34 kJ/mol for alpaca RBC. This suggests that in addition to the number of transport channels other features of the pathways might be important for defining the temperature dependence of the water permeability.     

ESR determination of rate constants for the radical polymerization of methyl phenethyl itaconate

Phenethyl 2-(methoxycarbonylmethyl)acrylate (methyl penethyl itaconate) ( 1 ) was prepared and its polymerization with dimethyl 2,2′-azoisobutyrate ( 2 ) was investigated kinetically in benzene. The polymerization rate (R_p) was found to be expressed by R_p = k·[ 2 ]^0,5·[ 1 ]^1,8 (at 50°C). Further, a higher dependence of R_p (2nd order) on the concentration of 1 was observed at 61°C. The overall activation energy of the polymerization was calculated to be a low value of 50,3 kJ/mol. The initiator efficiency (f) of 2 was determined to be 0,48 to 0,22 at 50°Cand 0,50 to 0,28 at 61°C. f decreases with increasing monomer concentration due to the high viscosity of 1 . The poly( 1 ) radical is stable enough to be observable by ESR at high temperatures (50–60°C). Rate constants of propagation (k_p) and termination (k_t) were estimated using the poly( 1 ) radical concentration determined by ESR. k_p [6,0 to 121/(mol·s) at 50°Cand 7,1 to 15 1/(mol·s) at 61°C] shows a trend to increase with the concentration of 1 . On the other hand, k_t [2,9·10⁴ to 17·10⁴1/(mol·s) at 50°Cand 6,9·10⁴ to 45·10⁴1/(mol. s) at 61°C] decreases with increasing MPI concentration. This behavior is responsible for the high order with respect to monomer concentration. Copolymerization of 1 (M₁) with styrene (M₂) at 50°Cgave the following results: r₁ = 0,36, r₂ = 0,40, Q₁ = 0,82 and e₁ = + 0,59. Using the above results, the rate constants of the cross-propagations were estimated to be k₁₂ = 22 1/(mol·s) and k₂₁ = 308 1/(mol·s) at 50°C.     

Macrophage handling of soluble immune complexes ingestion and digestion of surface-bound complexes at 4, 20 and 37 °C

Radioassays have been developed to measure, as separate events, the ingestion and degradation of macrophage-bound guinea pig IgG anti-DNP (2,4-dinitrophenyl)- DNP-BSA (bovine serum albumin) complexes of defined size and subclass. Complex ingestion was observed to be temperature-dependent and was effectively blocked only by a combination of inhibitors of respiration and glycolysis indicating that the process is under the same metabolic control as fluid phase pinocytosis. On the other hand, the half-life of membrane-bound complexes at 37 °C (t_1/2 = 5.6 ± 1.7 min) was considerably shorter than the value expected from membrane turnover studies (t_1/2 = 22.4 min) suggesting that complexes are selectively cleared from the cell surface. The rate of ingestion at 20°C was independent of complex size and of the IgG subclass used in complex formation, but was affected by in vivo stimulation of the macrophages before assay. Complex digestion was shown to be highly temperature-dependent and, at 37 °C, proceeded at a rate (t_1/2 = 1.5-5 h) which was 20-60-fold slower than the rate of ingestion indicating that the latter is unlikely to influence digestion kinetics. On the other hand, the selective action of 2-deoxyglucose in blocking digestion, but not ingestion, suggests that pinosome-lysosome fusion may play a part in determining the overall catabolic rate. A 2 to 3-fold difference in digestion rates was observed between the proteins employed as antibody (guinea pig IgG₁ or IgG₂) and as antigen (DNP-BSA). This finding suggests that the intrinsic susceptibility of ingested proteins to enzymatic hydrolysis may be the prime determinant of digestion rate. As with ingestion, no discrimination was observed in the degradation of complexes of different size or IgG antibody subclass. The observations in this and the preceding study (Eur. J. Immunol. 1980. 10: 317) indicate that complex size is important in determining the level of uptake by phagocytes but not subsequent events associated with catabolism.     

Kinetic and ESR studies of the radical-initiated polymerization of N-(2,6-dimethylphenyl)itaconimide

The polymerization of N-(2,6-dimethylphenyl)itaconimide (1) with azoisobutyronitrile (2) was studied in tetrahydrofuran (THF) kinetically and spectroscopically with the electron spin resonance (ESR) method. The polymerization rate (R_p) at 50°C is given by the equation: R_p = K [2] ^0,5 · [1] ^2,1. The overall activation energy of the polymerization was calculated to be 91 kJ/mol. The number-average molecular weight of poly (1) was in the range of 3500–6500. From an ESR study, the polymerization system was found to involve ESR-observable propagating polymer radicals of 1 under the actual polymerization conditions. Using the polymer radical concentration, the rate constants of propagation (k_p) and termination (k_t) were determined at 50°C. k_p (24–27 L · mol^?1 · s^?1) is almost independent of monomer concentration. On the other hand, k_t (3,8 · 10⁴–2,0 · 10⁵ L · mol^?1 · s^?1) increases with decreasing monomer concentration, which seems mainly responsible for the high dependence of R_p on monomer concentration. Thermogravimetric results showed that thermal degradation of poly (1) occurs rapidly at temperatures higher than 360°C and the residue at 500°C was 12% of the initial polymer. For the copolymerization of 1 (M₁) with styrene (M₂) at 50°C in THF the following copolymerization parameters were found; r₁ = 0,29, r₂ = 0,08, Q₁ = 2,6, and e₁ = +1,1.     

Comparative Nuclear Magnetic Resonance Studies of Diffusional Water Permeability of Red Blood Cells from Different Species. XII. Dog (<Emphasis Type="BoldItalic">Canis familiaris</Emphasis>) and Cat (<Emphasis Type="BoldItalic">Felis domestica</Emphasis>)

The diffusional water permeability (P _d ) of dog and cat red blood cell (RBC) membrane has been monitored by a doping nuclear magnetic resonance (NMR) technique on control cells and following inhibition with p-chloromercuribenzene sulphonate (PCMBS). The values of P _d were in the case of cat RBC ∼3.0 × 10⁻³ cm/s at 15 °C, 3.5 × 10⁻³ cm/s at 20 °C, 4.2 × 10⁻³ cm/s at 25 °C, 4.4 × 10⁻³ cm/s at 30 °C and 5.9 × 10⁻³ cm/s at 38 °C. In case of dog RBC the values of P _d were higher ∼3.8 × 10⁻³ cm/s at 15 °C, 4.6 × 10⁻³ cm/s at 20 °C, 5.0 × 10⁻³ cm/s at 25 °C, 5.9 × 10⁻³ cm/s at 30 °C and 7.9 × 10⁻³ cm/s at 37 °C. Systematic studies of the effect of PCMBS on water diffusion indicated that in the case of dog RBCs the maximal inhibition was reached in 15–30 min with 1 mm PCMBS, whereas in the case of cat RBCs in 60 min with 1 mm PCMBS or in 30 min with 2 mm PCMBS. The values of maximal inhibition in the case of cat RBC were in the range of 55–60% at 15 °C, 60–68% at 20 °C and 25 °C, 50–60% at 30 °C and 50–55% at 37 °C. In the case of dog RBC the corresponding values were higher, 75–80% at 15 °C, 70–80% at 20 °C and 25 °C, 65–70% at 30 °C and 55–60% at 37 °C. The basal permeability to water was estimated to be ∼1 × 10⁻³ cm/s −2 × 10⁻³ cm/s in the range of temperatures of 25–37 °C. The activation energy of water diffusion E _a,d was ∼19 kJ/mol for the dog RBC and ∼23 kJ/mol for the cat RBC. After incubation with PCMBS the values of E _a,d increased, reaching 40 kJ/mol in conditions of maximal inhibition of water exchange. The membrane polypeptide electrophoretic pattern of dog and cat RBCs has been compared with its human counterpart. Dog and cat RBCs contained higher amounts of spectrin (band 1 and 2) and lower amounts of bands 4.4, 4.2, band 5 and band 7 compared to human RBCs. Band 4.9 was decreased only in the cat RBCs, whereas band 6 was decreased only in the dog RBCs. Correspondence and offprint requests to: Gheorghe Benga, Department of Cell and Molecular Biology, ‘Iuliu Haţieganu’ University of Medicine and Pharmacy, 6 Pasteur St, 3400 Cluj-Napoca, Romania. Tel:/Fax: 40–64–194373; e-mail: GBenga@personal.ro; gbenga@umfcluj.ro     

Effect of tert‐Butyl Chloride on the Isoprene Polymerization with Neodymium Isopropoxide/Diisobutylaluminum Hydride and Neodymium Isopropoxide/Methylaluminoxane Catalysts

The effect of tert‐butyl chloride (tBuCl) as a third catalyst component on the isoprene polymerization with neodymium isopropoxide (Nd(OiPr)₃)/diisobutylaluminum hydride (Al(iBu)₂H) and Nd(OiPr)₃/methylaluminoxane(MAO) catalysts was examined in heptane at a fixed [Al]/[Nd] ratio of 30 and reaction temperatures of 30 and 60 °C. The Nd(OiPr)₃/Al(iBu)₂H catalyst was inactive without tBuCl, i.e., at [Cl]/[Nd] = 0, while it showed the highest activity under the conditions of [Cl]/[Nd] = 3.0 and an addition order of {Al(iBu)₂H + tBuCl} + Nd(OiPr)₃ to yield polyisoprene with relatively low molecular weight (M _n ca. 5 × 10⁴), broad MWD (M _w/M _n ca. 5.4), and high cis‐1,4 content (ca. 96%) at 30 °C. On the other hand, the Nd(OiPr)₃/MAO catalyst was effective at [Cl]/[Nd] = 0, and the most active at [Cl]/[Nd] = 1.0–1.5 with an addition order of {Nd(OiPr)₃ + MAO} + tBuCl to afford polymer with high molecular weight (M _n ca. 8 × 10⁵), fairly narrow MWD (M _w/M _n ca. 1.6) and high cis‐1,4 content (ca. 94%) at 30 °C. Thus, the novel Nd(OiPr)₃/MAO/tBuCl system features high activity, high molecular weight, narrow MWD, and high cis‐1,4 stereospecificity, as compared with the conventional Nd(OiPr)₃/Al(iBu)₂H/tBuCl system.

Effect of the [Cl]/[Nd] mole ratio on the cis‐1,4 contents with Nd(OiPr)₃/Al(iBu)₂H/tBuCl catalyst. Curve a: at 30 °C, 180 min; curve b: at 60 °C, 90 min.     

Kinetic studies of the anionic polymerization of methyl methacrylate in tetrahydrofuran with Na+ as counter ion,using monofunctional initiators

A newly designed automatically controlled stirred reactor suitable for kinetic measurements of reactions with half lives ≥2s has been applied to follow the anionic polymerization of methyl methacrylate in THF with Na⁺ as a counter ion in the presence of an excess of NaB(C₆H₅)₄. As initiators were used: benzylsodium reacted with α-methylstyrene (I), fluorenylsodium (II), and 9-methylfluorenylsodium (III). With I the initiation is fast as compared with the polymerization reaction which is first order in monomer concentration. Within the range of ?50°C to ?100°C an almost unperturbed “living” polymerization is observed. The Arrhenius plot of the rate constants results in a straight line with activation energy E_a = 4,4kcal·mol^?1 (= 18kJ·mol^?1) and frequency exponent A = 7,0.II and III are slow initiators, II giving rise to side reactions because of the “acidic” proton in 9-position after initiation, III exhibiting a rate constant of initiation k_i = 1l·mol^?1·s^?1 at ?72°C. The termination reaction is becoming increasingly important with increasing temperature and seems to be a unimolecular reaction with E_a,t = 11,5kcal·mol^?1 (= 48 kJ·mol^?1) and A_t = 10. Since the basic feature of the reactor is the possibility of drawing samples, polymers from each state of the reaction were available to be investigated also with respect to their tacticity. The monomer addition was shown to follow Bernoullian statistics. A structure of the “living” end being in harmony with the results observed is discussed.     

Kinetic and electron paramagnetic resonance studies on radical polymerization. Radical-initiated polymerization of methyl N-phenylitaconamate in N,N-dimethylformamide

The polymerization of methyl N-phenylitaconamate(methyl 2-methylenesuccinanilate ( 1 )) with dimethyl 2,2′-azodiisobutyrate ( 2 ) was studied in N,N-dimethylformamide (DMF) kinetically and by means of electron paramagnetic resonance (EPR) spectroscopy. The polymerization rate (R_p) at 55°C is given by the equation: R_p = k[ 2 ]^0,58 · [ 1 ]^1,6. The overall activation energy of the polymerization was calculated to be 54,2 kJ/mol. The number-average molecular weight of poly( 1 ) was in the range between 5000 and 17000. From an EPR study, the polymerization system was found to involve the EPR-detectable propagating polymer radical of 1 at practical polymerization conditions. Using the concentration of polymer radicals, the rate constants of propagation (k_p) and termination (k_t) were determined for 55°C. The rate constant of propagation k_p (between 8,4 and 12 L · mol^?1 · s^?1) tends to somehow increase with increasing monomer concentration. On the other hand, k_t (between 1,9. 10^?5 L · mol^?1 · s^?1) increases with decreasing monomer concentration, which results from a considerable dependence of k_t on the polymer-chain length. Such monomer-concentration-dependent k_p and k_t values are responsible for the high dependence of R_p on the monomer concentration. Thermogravimetric results showed that thermal degradation of poly( 1 ) occurs rapidly at temperatures higher than 200°C and the residue at 500°C amounts to 26% of the initial polymer. For the copolymerization of 1 (M₁) with styrene (M₂) at 55°C in DMF the following copolymerization parameters were found: r₁ = 0,52, r₂ = 0,31, and Q, e values Q₁ = 1,09 and e₁ = +0,55.     

Inflammation-induced synthesis of proteoheparan sulfate: A novel acute-phase reactant in rat hepatocytes

The synthesis of proteoheparan sulfate in hepatocytes is positively regulated under acute-phase conditions produced either by turpentine or deep back incision. In both cases the incorporation of [³⁵S]sulfate and [¹⁴C]glucosamine is doubled during a 4-h incubation period if compared with control rat hepatocytes. Neither the fractional secretion rate of heparan sulfate into the medium (< 0.1 of cell-associated glycosaminoglycans) nor the composition of newly formed proteoglycans in hepatocytes are affected during acute phase reaction.     

Study of photopolymer, 10. Cationic polymerization of vinyloxyethyl acrylate

Cationic polymerizations of vinyloxyethyl acrylate (VEA) catalyzed by boron trifluorideether complex were carried out in toluene at 10, ?10, and ?30°C. In this system, a critical catalyst concentration (C₀) exists; when the catalyst concentration is below this concentration, the polymerization of VEA cannot be initiated. The rate of polymerization is expressed as –d[M]/dt=k_p(C^*)·[M]^a, where a was found to be 0,44 (at 10°C), 0.51 (at ?10°C), and 0,63 (at ?30°C), respectively. From these results, it is concluded that the rate of polymerization of VEA is proportional to the first power of catalyst concentration and to 0,44–0,63 power of monomer concentration between 10 and ?30°C, respectively. Furthermore, it is recognized that an increasing monomer concentration tends to decrease the degree of polymerization.     

Comparative Cell Shape and Diffusional Water Permeability of Red Blood Cells from Indian Elephant ( Elephas maximus ) and Man ( Homo sapiens )

Red blood cells (RBC) from an Indian elephant (Elephas maximus) were studied by light microscopy (LM), scanning electron microscopy (SEM) and a new nuclear magnetic resonance (NMR) ‘imaging’ method based on the translational diffusion of water, NMR q-space analysis. Also, the transmembrane diffusional permeability, P _d of water in RBC was measured by using a Mn²⁺-doping NMR technique, taking human RBC as a reference. The main diameter of the elephant RBC was measured as 9.3 ± 0.7 μm by LM, 9.3 ± 0.7 μm by ‘shrinkage-corrected’ SEM, and 9.3 ± 0.4 μm by q-space anlaysis. The value is ∼1.4 μm larger than that for the human RBC. The values of P _d were, in the case of elephant RBC, 3.2 × 10⁻³ cm/s at 25 °C, 3.9 × 10⁻³ cm/s at 30 °C, 5.2 × 10⁻³ cm/s at 37 °C and 6.5 × 10⁻³ cm/s at 42 °C; all values were significantly lower than the corresponding values of P _d for human RBC, namely 4.3 × 10⁻³ cm/s at 25 °C, 5.2 × 10⁻³ cm/s at 30 °C, 6.1 × 10⁻³ cm/s at 37 °C, 7.8 × 10⁻³ cm/s at 42°C. The maximal inhibition of P _d (56%) was reached in 30 min at 37 °C with 2 mm p-chloromercuribenzene sulphonate (PCMBS) for both species of RBC. The basal permeability to water at 37 °C was estimated to be 2.3 × 10⁻³ cm/s for elephant and 2.6 × 10⁻³ cm/s for human RBC. The values of the activation energy for water permeability (E _a,d ) was significantly higher for elephant RBC (31.9 kJ/mol) than for human RBC (25.9 kJ/mol). This indicated that features other than the number of transporters per cell are likely to be important in defining the differences in water permeability in the RBC from the two species.     

Comparative Cell Shape and Diffusional Water Permeability of Red Blood Cells from Indian Elephant (Elephas maximus) and Man (Homo sapiens)

Red blood cells (RBC) from an Indian elephant (Elephas maximus) were studied by light microscopy (LM), scanning electron microscopy (SEM) and a new nuclear magnetic resonance (NMR) ‘imaging’ method based on the translational diffusion of water, NMR q-space analysis. Also, the transmembrane diffusional permeability, P _d of water in RBC was measured by using a Mn²⁺-doping NMR technique, taking human RBC as a reference. The main diameter of the elephant RBC was measured as 9.3 ± 0.7 μm by LM, 9.3 ± 0.7 μm by ‘shrinkage-corrected’ SEM, and 9.3 ± 0.4 μm by q-space anlaysis. The value is ∼1.4 μm larger than that for the human RBC. The values of P _d were, in the case of elephant RBC, 3.2 × 10⁻³ cm/s at 25 °C, 3.9 × 10⁻³ cm/s at 30 °C, 5.2 × 10⁻³ cm/s at 37 °C and 6.5 × 10⁻³ cm/s at 42 °C; all values were significantly lower than the corresponding values of P _d for human RBC, namely 4.3 × 10⁻³ cm/s at 25 °C, 5.2 × 10⁻³ cm/s at 30 °C, 6.1 × 10⁻³ cm/s at 37 °C, 7.8 × 10⁻³ cm/s at 42°C. The maximal inhibition of P _d (56%) was reached in 30 min at 37 °C with 2 mm p-chloromercuribenzene sulphonate (PCMBS) for both species of RBC. The basal permeability to water at 37 °C was estimated to be 2.3 × 10⁻³ cm/s for elephant and 2.6 × 10⁻³ cm/s for human RBC. The values of the activation energy for water permeability (E _a,d ) was significantly higher for elephant RBC (31.9 kJ/mol) than for human RBC (25.9 kJ/mol). This indicated that features other than the number of transporters per cell are likely to be important in defining the differences in water permeability in the RBC from the two species.     

Kinetic and ESR studies on radical polymerization. Radical polymerization behavior of N-octadecylmaleimide in benzene

The polymerization of N-octadecylmaleimide ( 1 ) initiated with azodiisobutyronitrile ( 2 ) was investigated kinetically in benzene. The overall activation energy of the polymerization was calculated to be 94,2 kJ·mol^?1. The polymerization rate (R_p) at 50°C is expressed by the equation, R_p = k[ 2 ]^0,6[ 1 ]^1,7. The homogeneous polymerization system involves ESR-detectable propagating polymer radicals. Using R_p and the polymer radical concentration determined by ESR, the rate constants of propagation (k_p) and termination (k_t) were evaluated at 50°C. k_p (33 L · mol^?1 · s^?1 on the average) is substantially independent of the monomer concentration. On the other hand, k_t (0,3 · 10⁴ – 1,0 · 10⁴ L · mol^?1 · s^?1) is fairly dependent on the monomer concentration, which is ascribable to a high dependence of k_t on the chain length of rigid poly( 1 ). This is the predominant factor for the high order with respect to the monomer concentration in the rate equation. In the copolymerization of 1 (M₁) and St (M₂) with 2 in benzene at 50°C, the following copolymerization parameters were obtained: r₁ = 0,11, r₂ = 0,09, Q₁ = 2,1, and e₁ = +1,4.