Platelet density and size: the interpretation of heterogeneity

Platelets have been separated according to buoyant density using a colloidal silica-polyvinylpyrrolidone system and subjected to electronic sizing. All density populations were found to be heterogeneous in size, the most dense platelets ranging from less than 3 fl to greater than 21 fl in both man and rat. Light platelet fractions contained no platelets greater than 13 fl in either species. Cohort labeling with [75Se]selenomethionine showed no indication of significant change in platelet buoyant density with ageing; greater specific activity found in young, dense platelets appears to be related to increased protein synthetic activity shown in vitro and likely to occur also in their precursor megakaryocytes. It is postulated that dense, intermediate and light platelets are released synchronously by the three different ploidy classes of megakaryocyte, that varying density indicates differing structural characteristics and presumably differences in function. The present findings do not deny the possibility that platelets decrease in size with ageing but if such occurs, it is not associated with a significant change in platelet buoyant density.     

Enhanced toxicity for mice of combinations of bacterial lipopolysaccharide and vincristine

Sublethal doses of vincristine (VNC) and bacterial lipopolysaccharide (LPS) administered simultaneously to adult male mice resulted in markedly enhanced mortality. All of 10 strains of Pseudomonas aeruginosa tested, 4 of 7 strains of Bacteroides, and 6 of 10 strains of Listeria monocytogenes were able to substitute for purified LPS in enhancing mortality in VNC-treated mice. Inoculation of mice with each of 10 strains of Pseudomonas, each of 7 strains of Bacteroides, and about half of the 10 strains of Listeria tested elicited increased resistance to the lethal action of purified LPS. The patterns of responses of mice receiving a lethal combination of 2 mg of LPS/kg and 1 mg of VNC/kg resembled those of mice receiving a lethal dose of 10 mg of VNC/kg alone or 15 mg of LPS/kg alone with respect to (i) serum glutamic pyruvate transaminase activity, (ii) hematocrit values, and (iii) thrombocytopenia. The patterns of responses of mice receiving a lethal combination of LPS and VNC resembled those of mice receiving a lethal dose of LPS alone with respect to (i) hypothermia, (ii) retention of sulfobromophthalein, (iii) fibrinogen level, (iv) prothrombin activity, (v) blood urea nitrogen levels, and (vi) time of death. These data are consistent with the proposition that the combination of VNC and LPS produces a fatal renal failure. Histological studies confirmed that there was extensive renal damage in mice treated with lethal doses of LPS alone or a lethal combination of LPS and VNC.     

Influence of lung volume and electrode position on electromyography of the diaphragm

In cats anesthetized with Nembutal, electromyograms of the diaphragm (Edi) were recorded from an anchored esophageal electrode, a pair of silver hooks inserted in the paratendinous region, and a pair of silver hooks and a pair of clips of small surface inserted in the costal region of the diaphragm facing the rib cage at FRC but covered with lung tissue at FRC + 80 ml. When single supramaximal electrical stimuli were applied to an isolated phrenic nerve, changes in lung volume from RV to near TLC had a negligible effect on muscle potentials from esophageal or paratendinous hooks, but increased the amplitude of potentials recorded from peripheral hooks and clips. In addition, it was found that small displacements of the esophageal electrode caused substantial changes in the amplitude of the recorded muscle potentials. The integration of the Edi spontaneously generated during occluded inspirations, recorded from paratendinous hooks and the esophageal electrode was linearly related to transdiaphragmatic pressure up to 50 cmH2O at all lung volumes. Above that level, esophageal electrode recordings showed a curvilinear Edi/Pdi relationship, while hook recordings showed a rectilinear relationship.     

Changes in bioactive compounds and antioxidant activity during convective drying of murta (Ugni molinae T.) berries

Murta (Ugni molinae T.) berries were air‐dried at five temperatures (40, 50, 60, 70 and 80 °C), and the changes in β‐carotene, phenolic acids, total phenolic and flavonoid contents and antioxidant capacities (DPPH and ORAC) were investigated. The berries showed a high content of β‐carotene, which decreased during drying temperature between 40 °C and 80 °C. Free and bound phenolic acids were also determined, showing gallic acid to be the prevalent phenolic acid. Total phenolic and flavonoid contents in the dried berries showed a higher decrease at lower temperature due to longer drying time. The radical‐scavenging activity also showed higher antioxidant activity at higher drying temperatures (70–80 °C) than at lower drying temperatures (40–50 °C). Total phenolic content (TPC) and flavonoids showed good correlation with antioxidant capacity. Murta berries proved to be an excellent source of antioxidants and bioactive compounds and are therefore a potential ingredient for new functional food products.     

Hot-air drying characteristics of Aloe vera (Aloe barbadensis Miller) and influence of temperature on kinetic parameters

The aim of this research was to study and model the kinetics of the hot-air drying of Aloe vera (Aloe barbadensis Miller) and to evaluate the influence of temperature on the kinetic parameters for the proposed models. A convective dryer was used at 50, 60, 70, 80 and 90 °C with an air flow of 2.0±0.2 m/s. The sorption isotherm of the fresh product was mathematically described by the Guggenheim, Anderson and de Boer (GAB) model, giving as a result monolayer moisture of 0.20 g water/g d.b. The mathematical models evaluated in the kinetic research included five empirical equations (Newton, Henderson-Pabis, Page, modified Page and Fick's diffusional model). The fit quality of the proposed models was evaluated by using the linear regression coefficient (r²), sum square error (SSE), root mean square error (RMSE) and Chi-square statistic (χ²). The diffusivity coefficient increased with the temperature from 5.30 to 17.73×10⁻¹⁰ m²/s, for a range of temperatures between 50 and 90 °C, with an estimated activation energy of 30.37 kJ/mol. When comparing the experimental moisture values with those estimated by the proposed models, the modified Page model provided the best to fit of the data, showing that this equation correctly simulates the Aloe vera drying process and represents an excellent tool for estimating its drying time.     

O6-alkylguanine-DNA alkyltransferase inactivation by ester prodrugs of O6-benzylguanine derivatives and their rate of hydrolysis by cellular esterases

To modulate the bioavailability and perhaps improve the tumor cell selectivity of O6-alkylguanine-DNA alkyltransferase (AGT) inactivators, pivaloyloxymethyl ester derivatives of O6-benzylguanine (BG) were synthesized and tested as AGT inactivators and as substrates for cellular esterases. The potential prodrugs examined were the 7- and 9-pivaloyloxymethyl derivatives of O6-benzylguanine (7- and 9-esterBG), and of 8-aza-O6-benzylguanine (8-aza-7-esterBG and 8-aza-9-esterBG) and the 9-pivaloyloxymethyl derivative of 8-bromo-O6-benzylguanine (8-bromo-9-esterBG). The benzylated purines were all potent inactivators of the pure AGT and of the AGT activity in HT29 cells and cell extracts. Each ester was at least 75 times less potent than the corresponding benzylated purine against the pure human AGT. In contrast, the activities of esters and their respective benzylated purine were similar in crude cell extracts and in intact cells. The increase in potency of esters in cellular extracts could be explained by a conversion of the respective prodrug to the more potent benzylated purine in the presence of cellular esterases. The apparent catalytic activity (Vmax/Km) of liver microsomal esterase for 8-azaBG ester prodrugs was 70-130 times greater than for BG prodrugs and 10-20 times greater than for 8-bromo-9-esterBG. Tumor cell hydrolysis of the esters varied considerably as a function of cell type and prodrug structure. These data suggest that these or related prodrugs may be advantageous for selective AGT inactivation in certain tumor types.