Use of triple stain technique for simultaneous assessment of vitality and acrosomal status in boar spermatozoa

The object of this study was to adapt the triple stain technique to diluted and incubated boar spermatozoa. Freshly ejaculated semen was resuspended in MR-A diluent to contain 3x10(7) cells/ml (diluted spermatozoa) and was subsequently capacitated (incubated spermatozoa). Experiments were conducted to show the conditions required for optimal staining quality and validation of triple stain technique. The most satisfactory staining solutions for diluted spermatozoa were 2% Trypan blue at 37 degrees C for 15 minutes, 0.8% Bismarck brown in 30% ethyl alcohol (pH 2.8) at 40 degrees C for 10 minutes and 0.8% rose Bengal in 0.1 M of Tris (pH 4.3) at 21 degrees C for 20 minutes. Satisfactory results were obtained for incubated spermatozoa with rose Bengal when the staining time was 10 minutes. Triple stain technique seemed to be a useful method for the simultaneous assessment of sperm vitality and acrosomal status; consequently, it should be valuable tool, for use in porcine in vitro fertilization systems.     

Construction and properties of K1 type killer wine yeasts

Summary With the use of a protoplast fusion technique the killer character of K1 type was transferred into four industrial Saccharomyces wine yeasts. The prototrophic yeast strains active against standard sensitive and K2 killer Saccharomyces strains, resistant to K1 killer toxin were constructed with no changes in technological properties.     

The structural stability of an expression plasmid bearing a heterologous cloned gene depends on whether this gene is expressed or not

Summary During subculturing of recombinant Escherichia coli under conditions blocking cloned gene expression the plasmid was structurally stable for 200 generations. However, under conditions permitting foreign gene expression only 10% of plasmid containing cells had this gene functional after 100 generations.     

Mechanisms towards compensation of long-term haemopoietic injury in mice after 5 Gy irradiation:In vivo andin vitro enhancement of superoxide anion production by granulocytes

This paper analyzes the long-term (6 and 12 months) function of mouse granulocytes after total body irradiation with a single dose (5 Gy) of X-rays. Superoxide anion production has been investigated in granulocytes from peripheral blood, and also in those harvested from long term bone marrow cultures, with the aim of correlating the environmental damage induced by radiation with the functional properties of granulocytes. Anin vivo andin vitro enhancement of superoxide anion production and protein levels in granulocytes from irradiated mice is described. The presence of some colony stimulating factor in the supernatant of cultures from irradiated mice could play an important role in the priming of granulocytes.     

Modulation of the affinity of the vasopressin receptor by magnesium ions.

The binding of [3H]vasopressin (AVP) and the 125I-labelled vasopressin antagonist (VP-AT) d(CH2)5[Tyr2(Me),Tyr9(NH2)]AVP to rat liver membranes was examined with or without the addition of milimolar concentrations of divalent cations. The binding of vasopressin was enhanced by Mg2+ and Co2+ and markedly decreased by EGTA. The addition of EGTA and Mg2+ together restored the binding to a value similar to that of Mg2+ alone. On the contrary, the addition of Mg2+, Co2+, EGTA, and the combination of EGTA and Mg2+ decreased the binding of VP-AT to rat liver membranes. Kinetic analyses showed that Mg2+ increased the Kd twofold for VP-AT; that is from 0.13 nM to 0.28 nM. Moreover, it showed that the receptor with or without the addition of Mg2+ consists of a single population of binding sites, indicating that the receptor is switched from a high affinity to a low affinity state for VP-AT in the presence of 10 mM Mg2+. GTP gamma S was unable to block the effect of Mg2+ on the binding of VP-AT. These results suggest that this divalent cation interacts with receptor itself producing a conformational changes which thus modulates the affinity of the receptor.     

Histological characterization of in vitro regenerated structures of Panax ginseng

Callus was induced from mature leaf lamina, petiole, stem, and main and branch root of Panax ginseng and maintained under different nutrient and light conditions. Heterogenous callus cultures differentiated organoids and/or embryoids. Embryoids that arose from callus cultures of leaf origin germinated into shoots. The occurrence of the early stages of different regenerated structures was proved on histological level.     

Sex, light and carotenes: the development of Phycomyces.

Phycomyces blakesleeanus is known for the elaborate behaviour of its sporangiophores (fruiting bodies). Sporangiophore development is exquisitely sensitive to blue light, easy to describe quantitatively, pliable to genetic and biochemical research, and reminiscent in many details of other photoresponses in the same and in other organisms. The developmental and behavioural processes of Phycomyces share a number of genes. A combinatorial use of gene expression appears to be the basis for the complexities observed in this fungus.     

Conditions for selective degradation of lignin by the fungus Ganoderma australis

Summary The white-rot fungus Ganoderma australis selectively degrades lignin in the ecosystem palo podrido. Using conditions that simulate those of palo podrido in the laboratory, it was found that low nitrogen content and low O₂ tension stimulate the productio of manganese peroxidase and lignin degradation, and depress cellulose degradation and cellulase production. The inverse is found at high nitrogen concentration and high O₂ tension. This agrees with previous results indicating that low O₂ tension and low nitrogen stimulate selective lignin degradation by this fungus. Correspondence to: J. Eyzaguirre