External factors involved in the regulation of an extracellular proteinase synthesis in Bacillus megaterium

Summary A mathematical model was formulated to describe the kinetics and stoichiometry of growth and proteinase production in Bacillus megaterium. Synthesis of the extracellular proteinase in a batch culture is repressed by amino acids. The specific rate of formation of the enzyme (r _E) can be described by the formula {ie373-1}, where k ₂ and k ₃ stand for the non-repressible and repressible part of enzyme synthesis respectively, k _{S 2} is a repression coefficient and S ₂ indicates the concentration of amono acids; the values of k ₂ and k _{S 2} depend on the composition of the mixture of amino acids. Even in a high concentration, a single amino acid is less effective than a mixture of amino acids. The dependence of the proteinase repression on the concentration of an external amino acid (leucine) follows the same course as its rate of incorporation into proteins, approaching saturation at concentrations higher than 50 M (half saturation approximately 10 M). However, the total uptake of leucine did not exhibit any saturation even at 500 M external concentration.Symbols X biomass concentration, g/l - E proteinase concentration, unit/l - t time, h - S ₁ concentration of glucose, g/l - S ₂ concentration of amino acids, g/l - specific growth rate, l/h - rE specific rate of enzyme production, unit/g/h - k ₁ growth kinetic constant, l/h - k ₂ product formation kinetic constant (for non-repressible part of enzyme synthesis), unit/g - k ₃ product formation kinetic constant (for repressible portion of enzyme synthesis), unit/g - k _{S 1} saturation constant, g/l - k _{S 2} repression coefficient for a certain amino acid or amino acids mixture, g/l     

Characterization of specific HLA-DQ alpha allospecificities by genomic, biochemical, and serologic analysis

Bgl II restriction endonuclease digestion of genomic DNA from lymphoblastoid cell lines homozygous for HLA DR and DQ serological specificities, followed by hybridization with a DQ alpha cDNA probe, identified a genomic polymorphism characterized by two reciprocal patterns, one associated with DR 3, 5 and 8 and the other with DR 1, 2, 4, 7, and 9. The former pattern corresponded precisely to the reactivity of monoclonal antibody SFR20-DQ alpha 5, shown by Western blotting to react with isolated alpha-chains, but not with beta-chains. Additional variants of the DQ alpha genes were identified by using a locus-specific oligonucleotide probe for the DQ alpha gene, indicating differences among the DQ alpha 5-negative set of alleles. This analysis defines a set of DQ alpha allelic markers that are distinct from the well-established DQ serologic specificities DQw1, 2, 3 or "blank." Although most DQ alpha 5+ cells carry the DRw52 specificity associated with the DR beta 2 gene, analysis of DQ alpha polymorphisms on DR5, DQw1; DR8, DQw1; and DRw13, DQw1 cells verified that this DQ alpha family of alleles was not invariably linked to the DR beta 2 locus.     

Autologous B lymphoblastoid cell lines and long-term cultured T cells as stimulators in the mixed lymphocyte reaction: analysis of the role of HLA class II antigens as stimulatory molecules

Human activated T cells, long-term cultured in the presence of interleukin 2 (IL 2), were compared with autologous Epstein Barr virus-transformed B lymphoblastoid cell lines for expression of human leukocyte (HLA)-HLA-DR and -DQ antigens and for ability to induce proliferative responses in autologous and allogeneic lymphocytes. Immunofluorescence analysis performed with a panel of monoclonal antibodies (mAb) specific for HLA-DR or -DQ antigens did not reveal any significant difference in the expression of HLA-DR antigens but revealed reduced expression of HLA-DQ antigens on two out of four T cell lines tested. No obvious difference could be detected in the two-dimensional gel electrophoretic profile of HLA-DR and -DQ beta-chains synthesized by the autologous pairs of B and T cell lines. In contrast with previous reports, the IL 2-dependent cell lines consistently induced alloproliferative responses in standard 6-day mixed lymphocyte cultures; however, these responses were severalfold lower than those elicited by the autologous B lymphoid lines. Both anti-HLA-DR and anti-HLA-DQ mAb blocked the proliferative responses induced by the B cell lines but did not affect those generated by the T cell lines, suggesting that the latter cells induce T lymphocyte activation via a mechanism independent of HLA-DR or -DQ antigen expression on their surface. Addition of IL 2 to the mixed cultures with B cell lines as stimulators did not affect the outcome of the proliferative responses but partially or completely reversed the blocking activity of the mAb. In contrast, IL 2 significantly enhanced the alloproliferation induced by the T lymphoblastoid cell lines, and the anti-HLA class II mAb partially antagonized this effect. Taken together, these data suggest that unlike the HLA-DR and -DQ gene products on B cells, those on IL 2-dependent long-term cultured T cells do not play a direct or primary stimulatory role in the mixed lymphocyte reaction; the reduced levels of alloproliferation induced by the T cell lines are, at least in part, due to a defective production of endogenous IL 2 by the responder lymphocytes rather than to a defective expression of IL 2 receptors by the alloproliferative T cell subset; and the anti-HLA class II mAb in these cultures act only at the responder cell level, since they can efficiently block the enhancement of T cell proliferation triggered by exogenous IL 2, but not the proliferative responses induced by T cell lines in standard conditions.     

Significance of various nitrogen sources for calcium deficient seedlings ofCucurbita pepo L.

The aim of the present investigation was to examine whether some of the growth defects of calcium-deficient pumpkin (Cucurbita pepo L.) seedlings are due to an incapability of the plants to assimilate nitrate nitrogen, such being the mechanism by which according to some of the previous authors growth defects in calcium deficient plants may be produced. The results of the present study, however, demonstrate that the accessibility of various possible products of nitrate reduction is not the limiting factor responsible for growth symptoms of calcium deficiency.     

Occurrence of lysosomes and the differentiation of rat liver cells

Summary The occurrence of lysosomes has been investigated electron microscopically and cytochemically in cells of rat liver in the course of ontogenesis.It has been found that primary lysosomes occur during the whole period under investigation and that they originate from the Golgi complex. Some of them assume the appearance of multivesicular bodies. Acid phosphatase activity is lower at the prenatal stage than after the birth. The occurrence of secondary lysosomes proceeds in two stages. Secondary lysosomes appear in a high number at the beginning of differentiation of the liver diverticulum (10–12 day of embryonic life). On the subsequent days they are, with few exceptions, no more present. At the end of the embryonic period (starting with the 20th day) and especially after the birth, they progressively grow in number and move from the region of central cytoplasm peripherally towards the bile capillary.Differences in occurrence of secondary lysosomes are in connexion with reconstruction of the liver primordium at the beginning of liver development and with the change in metabolism of the liver cell after the birth.     

Postnatal differentiation of the Golgi apparatus and the dendrites of Purkinje cells of the rat cerebellum

Summary The morphology of postnatal differentiation of the Golgi apparatus, the nucleus, the perikaryon, and the dendrites was studied in Purkinje cells of the rat cerebellum for 30 days after birth using histochemical, histological, and electron microscopic methods.The Golgi apparatus during differentiation undergoes morphological and positional changes. From the 1st to 7th postnatal day, the Golgi apparatus is found in a supranuclear position, and is connected with the axes of differentiating primary dendrites by beam-like processes. From days 8 to 11 this connection disappears, and most of the Golgi apparatus assumes a lateronuclear and infranuclear position. After the 11th or 12th day, the Golgi apparatus is found in perinuclear and peripheral cytoplasmic positions. The formation of granular endoplasmic reticulum occurs in the vicinity of the perinuclear Golgi apparatus. The differentiation of cell and nuclear forms requires approximately 20 days. The morphological changes of differentiation are discussed in relation to the participation of the Golgi apparatus in the differentiation of dendrites and in the formation of the granular endoplasmic reticulum.     

Peroxidases of different parts of the pumpkin plant (Cucurbita pepo L.)

We compared the occurrence of peroxidase isozymes in protein extract from roots, hypocotyls and cotyledons of 10 dayCucurbita pepo plants and of adult leaves of older plants by means of starch gel and polyacryl amide gel electrophoresis. We reached maximum discrimination by means of starch gel electrophoresis: 11 zones were ascertained on the cathode side and about 2 on the anode side at pH 3.1. Two zones occurred regularly:A and (the latter having a more complicated structure). ZoneD is characteristic for roots, but is it suppressed and seldom found with leaves. On the other hand zonesC ₁ andC ₂ are clearly discernible with leaves but are substantially less evident with roots. The character of anodic zoneZ is discussed later in this paper.     

The effect of physical conditions of cultivation on the respiratory metabolism of algae

V práci jsem zji?tovala, zda podmínky kultívace ?as ovlivńují ?espira?ní metabolismus. Rasy Chlorella pyrenoidosa (82), Scenedesmus obliquus (125) a Euglena gracilis (259) byly pěstovány ve t?epané a stojaté kultu?e. T?epáním kultur je podstatně ovlivněn respira?ní metabolismus ?as. T?épané kultury mají na rozdíl od stojatých sní?enou spot?ebu O₂ a vět?inou odli?né RQ. Je mo?né, ?e zji?těné rozdíly jsou podmíněny zrychleným vývojem a stárnutím t?epaných kultur. Je tedy t?epání jako zp?sob kultivace významným faktorem, který je nutno respektovat p?i pěstování experimentálního materiálu. Namě?ené hodnoty respira?ního kvocientu okolo 1,3 svěd?í pro to, ?e anaerobní glykolytické pochody mohou probíhat i za dokonalého p?ístupu vzduchu do media. Kultura Scenedesmus obliquus (125) má pravděpodobně málo p?izp?sobivý metabolismus a na změny prost?edí nereaguje tak citlivě jako Euglena gracilis nebo Chlorella pyrenoidosa.     

Production of taxanes by Taxus baccata plant cells

In callus cultures of Taxus baccata grown on agar media according to Murashige and Skoog supplemented with different growth hormones 8 taxol analogues were identified.     

Über den anaeroben Stoffwechsel der Vegetationskegel des Weizens

Na vegeta?ních vrcholech ozimé p?eniceTriticum vulgare var.milturum Al., odr?dy Chlumecká 12 jsme sledovali papírovou chromatografií změny v obsahu glycid?, aminokyselin a organických kyselin po inkubaci za aerobních a anaerobních (N₂) podmínek. Práce byly prováděny jak s isolovanými vegeta?ními vrcholy, tak s vrcholy in situ ve 2., 3. a 4. etapě organogenese. Také byl histochemicky stanoven obsah acetaldehydu, lokalisace a aktivita alkoholdehydrogenásy. Získané údaje svěd?í o tom, ?e v mladých vegeta?ních vrcholech (2. etapa organogenese) p?eva?uje aerobní kva?ení, kde?to u star?ích se ji? uplatňuje p?evá?ně aerobní oxydasové systémy. Na vegeta?ních vrcholech nedochází k syntese vět?iny sledovaných aminokyselin ve vět?ím mě?ítku, i kdy? jím byl poskytnut zdroj aminodusíku. Hlavním zdrojem pro proteosyntesu jsou aminokyseliny p?iváděné z list?, je v?ak mo?no uva?ovat o jejich p?eměnách ve vegeta?ním vrcholu. Tyto p?eměny probíhají jen za aerobních podmínek. Pro kontrolu, zda se aminokyseliny neuvolňují p?i proteolyse, byl během inkubace m ?en kapkovou metodou obsah bílkovin. P?edlo?ená práce bude dále doplněna p?ímým mě?ením aktivity některých enzym?.