Cloning,sequence analysis and yeast expression of the egl1 gene from Trichoderma longibrachiatum

A gene (egl1) encoding an endoglucanase (EGL1) from Trichoderma longibrachiatum has been cloned and sequenced. This gene, homologous to the T. reesei egl1 gene, differs from it in the length of the introns (particularly the first one) and encoded protein. A cDNA fragment obtained by the rapid amplification of cDNA ends method, which takes advantage of the polymerase chain reaction, has been expressed in yeast under control of the cyc-gal inducible promoter and yeast clones able to secrete active enzyme have been obtained. Correspondence to: J. A. Pérez-González     

An energy-dependent efflux system for potassium ions in yeast

An efflux of potassium ions was demonstrated in mutants of yeast cells lacking a functional high affinity carrier system for monovalent cations. This efflux showed the following characteristics: (a) It was stimulated by the presence of a substrate, either glucose or ethanol. (b) It was stimulated by several cationic organic molecules, such as ethidium bromide, dihydrostreptomycin, diethylaminoethyldextran, and also by trivalent cations, such as Al3+ and lanthanides; this stimulation also depended on the presence of a substrate. (c) K+ efflux was decreased in yeast mutants with decreased ATPase activity, which generated a lower membrane potential. (d) Although the efflux appeared to be of an electrogenic nature, producing hyperpolarization of cells, it was accompanied by the efflux of phosphate, probably as an anion partially compensating for the large amount of cations leaving the cell. (e) K+ efflux was also accompanied by an uptake of protons. (f) The efflux appeared more clearly in cells grown in YPD medium, and not in more complex media nor in the same YPD medium if supplemented with Ca2+ or Mg2+. Efflux of monovalent cations produced by Tb3+ and organic cationic agents was also demonstrated in wild type strains. This efflux system appears to be, at least partially, electrogenic, but seems to be also an exchange system for protons and to function as a symport with phosphate; it may be involved in the regulation of the internal pH of the cell, and appears to be regulated by its link to the energetic status of the cell, probably through the membrane potential.     

Bovine colostrum or milk as a serum substitute for the cultivation of a mouse hybridoma

A mouse-mouse hybridoma was grown in serum-free medium supplemented with bovine milk or colostrum. Bovine colostrum supported growth of the hybridoma whereas bovine milk alone did not support cellular proliferation. For growth in medium supplemented with colostrum, the maximum cell concentration achieved was 1.4 x 10(6) cells/mL in 2.2% colostrum, which is 44% of that obtained in 9% serum. When cells were grown in media containing milk and low amounts of serum (<1%) the maximum cell concentration in 2.2% milk with 0.4% serum was 2 x 10(6) cells/ml, whereas it was only 0.2 x 10(6) cells/ml and 1.3 x 10(6) cells/ml in 2.2% milk alone and 0.4% serum alone, respectively. Similar behavior was observed for growth in media containing colostrum and low amounts of serum. The monoclonal antibody production in media containing combinations of serum and milk or colostrum was comparable to that obtained in media with higher serum concentrations. Experiments performed with conditioned media suggest that the rapid decrease in viability, after the maximum cell concentration has been reached, is partially due to the presence of some inhibitory components generated during the cell culture rather than due to depletion of some serum components.     

Biochemical changes in the germinating seeds ofTrigonella foenum graecum L. in relation tos-triazine herbicides

Thes-triazine herbicides studied (atrazine and simazine) inhibited the growth of the seedlings ofTrigonella foenum graecum L.(Viciaceae). These herbicides caused a decrease in the contents of total water soluble carbohydrates and reducing sugars. Pentose sugars increased in the cotyledons. The total protein and free amino acid contents were enhanced. The activity of peroxidase was inhibited in the cotyledons, but was enhanced in the seedling axes.     

Interaction of ethidium bromide with the transport system for monovalent cations in yeast

Summary Ethidium was found to be taken up by yeast cells in a process that, at certain concentrations has the main following characteristics: a) a substrate is required; b) it presents cooperative kinetics, withn, according to the Hill equation 3; c) ethidium can be concentrated more than 100-fold; d) the uptake is inhibited by Ca²⁺; e) the uptake of the dye is inhibited by monovalent cations with a selectivity pattern similar to that observed in their transport by yeast; f) ethidium inhibits the uptake of K⁺, and, at concentrations up to about 250 m produces a competitive inhibition on the uptake of Rb⁺; and g) ethidium produces the same effects as K⁺ on respiration and the extrusion of H⁺. It is concluded that ethidium is taken up by yeast cells in a selective way by the same transport system normally employed for monovalent cation uptake.     

New germacranolides from Liatris species

Three closely related cis-Δ⁴-diepoxygermacranolides were isolated from two Liatris species. Structures of eleganin from L. elegans (Walt.) Michx, and liscundin and liscunditrin from L. secunda (Ell.) Small. were established by NMR spectrometry, chemical transformations and correlation with punctaliatrin previously isolated from L. punctata Hook. Reinvestigation of the latter species yielded another germacranolide liatripunctin whose structure is reported.     

A possible cause of non-disjunction of additional chromosome 21 in Down syndrome

Summary A possible cause of non-disjunction of chromosome 21 in Down Syndromes has been cytogenetically evaluated by examining the parents by Ag-staining technique. In all the cases studied so far, the contributing parents have active ribosomal cistrons on both chromosomes 21 i.e. both chromosomes are stained positively by silver staining. These results show that the active NORs might play an essential role in meiotic non-disjunction. Furthermore, the preliminary results demonstrate that the acrocentric associations of homologous and non-homologous nature involving chromosome 21 are the most frequent in the contributing parent which may further indicate the role of multiple cellular factors affecting the associations in promoting the non-disjunction in addition to active NORs. The possible mechanisms regarding the non-disjunction of chromosome 21 have been described.Presented at the 34th Annual Meeting of the American Society of Human Genetics, Norfolk, VA, USA