The use of acridine orange for testing blood platelet integrity

Two processes are involved in the accumulation of acridine orange in human blood platelets. One follows a diffusion like kinetics and is independent of the ATP level whereas the second one can be completely abolished by ATP depletion. The acridine orange incorporation rate seems to be a suitable parameter for testing platelet integrity. It reflects very sensitively the influence of the preparation method as well as of anticoagulating substances used on the stability of platelet suspensions. The rates of acridine orange incorporation and of aggregation were measured in platelet-rich plasma and in saline suspended platelets after gel filtration, respectively, over a period of 120 min storage. Both rates are influenced to a different degree by anticoagulating agents such as citrate, heparin and EDTA. When contact with anticoagulating agents during platelet preparation is avoided, platelets show a constant acridine orange incorporation and aggregation during storage and the smallest morphological alteration.     

A method to identify individual proteins in four different two-dimensional gel electrophoresis systems: application to Escherichia coli ribosomal proteins.

Agarose gel electrophoresis offers a versatile means to fractionate nucleic acids varying in size over considerably different molecular weight ranges. Surface cohesion properties of agarose gels and sample loading problems have hampered the use of such gels in largediameter, preparative-scale tube gel electrophoresis. We report here a procedure that makes routine and reproducible the construction, sample loading, and running of preparative agarose electrophoretic gels. Data are presented on the fractionation of yeast nucleic acids.     

Copulatory vocalizations of female macaques (Macaca fascicularis): Variability factors analysis

In 234 copulations, male and female behavioural patterns were collected, especially the vocal pattern for the female. Moreover quantitative structural analysis was performed on 38 of these female copulatory vocalizations. A multifactorial analysis, ANAFAC, was performed to seek relations between (1) utterance of female copulatory call and male and female copulatory behaviour and (2) these patterns and the great variability of female calls. Utterance of female copulatory vocalizations is essentially due to the female’s own copulatory behaviour in periods of intense sexual activity. Long female calls are related to male ejaculation, and those “rich” in harmonics are related to visual and tactile communicatory signals exchanged by the male and the female during a mount. This analysis strongly confirms the tied bond between quality of a call and internal state of the emitter. Long calls, rich in harmonics, may serve as signals maintaining or strengthening the possibly temporary preferential relation between male and sexually receptive female.     

Correction of thermal gradient errors in stem thermocouple hygrometers

Stem thermocouple hygrometers were subjected to transient and stable thermal gradients while in contact with reference solutions of NaCl. Both dew point and psychrometric voltages were directly related to zero offset voltages, the latter reflecting the size of the thermal gradient. Although slopes were affected by absolute temperature, they were not affected by water potential. One hygrometer required a correction of 1.75 bars water potential per microvolt of zero offset, a value that was constant from 20 to 30 C.     

Ionomycin-mediated calcium transport in rigid and fluid liposomes

The antibiotic ionophore ionomycin translocates Ca from an aqueous medium into or across an organic immiscible phase. At pH 8.0, ionomycin translocates less Ca than A23187, the effects of these ionophores being additive to one another. The capacity of ionomycin to translocate Ca across the organic phase is dramatically decreased when the pH of the aqueous media is reduced from 8.0 to 7.5 or lower values. Ionomycin also mediates Ca exchange-diffusion in liposomes, the magnitude of such a process being greater in fluid than in rigid liposomes. At a physiological pH (7.4), ionomycin is unexpectedly as potent as A23187 in mediating Ca transport in fluid liposomes. These findings suggest that the capacity of ionophores to translocate Ca across model membranes depends on both the transverse and lateral mobility of the ionophoretic molecules. The relative importance of the latter phenomenon itself largely depends on the stoichiometry of the Ca-ionophore complex.     

Metabolisme lipidique chez l'Aspergillus versicolor (Vuill.) tiraboschi. Relations avec la biogenese de la sterig matocystine

Résumé L' Aspergillus versicolor est cultivé sur un milieu synthétique pendant 22 jours. Les productions de lipides et de stérigmatocystine sont comparées. Les acides gras des fractions neutres et polaires sont essentiellement: C 160, C 180, C 181 C 182, et C 183. Les quantités maximales de mycélium sec, de lipides neutres et de stérigmatocystine apparaissent respectivement au 4e, 7e et 20e jours. Une diminution de la teneur en lipides précède la phase de concentration maximale en métabolites secondaires du type polycéto-acide. Il semble que les lipides, et tout particulièrement l'acide palmitique, participent à la biogenèse de ces dérivés.

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Summary Aspergillus versicolor is cultivated in a synthetic medium for 22 days. Bioproduction of lipids and sterigmatocystin are compared. The fatty acids of the neutral lipid and polar lipids fractions are mainly: C 160, C 180, C 181, C 182, C 183. Maximal yields of dry weight, neutral lipids and sterigmatocystin occur, respectively, on the 4th, the 7th and the 20th days. These results and their comparison with other works emphasize that a fall of concentration in lipids precedes the phase of highest concentration in secondary metabolites of polyketide type; it appears that fats and particularly palmitic acid are present in biogenesis of these derivatives.

     

Purification and properties of mannitol dehydrogenase from Agaricus bisporus sporocarps

Mannitol dehydrogenase (mannitol: NADP⁺ 2-oxidoreductase: EC 1.1.1.138) was isolated from Agaricus bisporus by fractionation with protamine sulphate and (NH₄)₂SO₄, followed by chromatography on DEAE-Sephadex, then by affinity and gel chromatography. The products of enzyme reaction were identified by GLC and TLC. K_m, optimum pH, MW and pI of the enzyme as well as the influence of temperature, ions and inhibitors on enzymic activity were determined. In the sugar reducing reaction, the enzyme was specific for fructose but, in the reverse direction, some structurally related polyols could substitute for mannitol. The enzyme was very sensitive to alterations in the NADP⁺/NADPH ratio. The results are discussed in relation to the possible role of mannitol dehydrogenase in fungal metabolism.     

Dependence of the early steps of chlorophyll(ide) synthesis on respiration in dark-grown Euglena gracilis

Protochlorophyll(ide) disappearance and chlorophyll(ide) accumulation, in dark-grown Euglena, promoted by series of actinic light flashes, have been followed by in vivo fluorescence measurements. The data show that chlorophyll(ide) accumulation is biphasic, i.e., there is an initial rapid phase followed by a slower linear phase. The linear phase is highly dependent on flash frequency and on cell respiration whereas the initial phase is much less affected by these factors. It is concluded that dark-grown cells contain a limited pool of phototransformable protochlorophyll(ide); once this pool is exhausted, its reformation and/or the synthesis of some unknown metabolite necessary for the photoreduction appears to be dependent on respiration.     

Genetic control of igG2a production in the response to sheep erythrocytes

A low IgG2a response in B10 mice during the primary response to sheep red blood cells (SRBC) is described. Analysis of the response in B10 × BALB/c hybrid progenies and in congenic strains indicates that this low response is a dominant phenotype placed under the control of a single Mendelian gene or a group of closely linked genes. This gene(s) is neither linked to CH allotypes orH2 haplotypes, nor is it sex-linked. It can be considered as an isotype- and antigenspecific regulatory gene of the immune response.     

High resolution nuclear magnetic resonance studies of nigeran

Polymer motion in solution can be studied by ¹³CNMR relaxation methods, which provide information about the correlation time for C-H vectors. ¹³C-Relaxation and Nuclear Overhauser Enhancement (NOE) data may frequently be combined to determine the dipole-dipole relaxation contribution. An alternative method is proposed based on a comparison of the proton spin-lattice relaxation rates of the centre proton resonances of an unlabelled molecule with the relaxation rates of the ¹³C satellites (from ¹³C labelled molecules).Selectively labelled nigeran which is an alternating $\text{1 → 3 and 1 → 4 α-}\text{d}\text{-glucan}$ has been investigated. The discussion in terms of the occurrence of different motions for each of the two units of the polymer requires an unambiguous assignment of the two anomeric carbons. For this reason a detailed assignment of the ¹H and ¹³C Nuclear Magnetic Resonance (NMR) spectra of nigeran in dimethylsulphoxide-d₆ is described, based on T₁ and NOE measurements in addition to selective homonuclear and heteronuclear spin decoupling experiments. These values are correlated with a conformation estimated by HSEA hard-spheres calculation. The measurements of the relaxation parameters for labelled and unlabelled compounds which provide an alternative determination of the ¹³C-¹H dipole-dipole relaxation contribution in a macromolecule agree well with ¹³C-{¹H} NOE experiments.