Effects of cultivation techniques and media on yields and morphology of the basidiomycete Armillaria mellea

Summary The yield of cell mass and the morphology of Armillaria mellea, strain ATCC 11114, was studied using a variety of cultivation methods: solid media, standing liquid culture, shake flasks, tower reactors and impeller-stirred reactors. Two different media, malt extract broth and a glucose/asparagine/peptone-medium, and the corresponding agar media, were used. Yields were higher in the malt extract media than in the glucose media. Generally the highest yields were obtained on solid media while agitated cultures gave the lowest yields. Morphological characteristics such as pellet formation, adhesion to surfaces and pigment production were significantly affected by culture conditions.     

Sequence of the mglB gene from Escherichia coli K12: comparison of wild-type and mutant galactose chemoreceptors

Summary The mglB gene of Escherichia coli codes for a galactose-binding protein (GBP) that serves both as the galactose chemoreceptor and as the recognition component of the -methylgalactoside transport system. The mglB551 mutation eliminates the chemotactic function of GBP without altering its transport or substrate-binding properties. To investigate the interaction between GBP and Trg, the chemotactic signal transducer for galactose, we sequenced the mglB genes from wild-type and mglB551 mutant strains. The mutation causes the replacement of Gly₇₄ of GBP by Asp. This residue is located in alpha-Helix III at the tip of the P domain in the GBP tertiary structure farthest removed from the substrate-binding cleft between the P and Q domains. We conclude that Helix III must be part of, or at least adjacent to, the recognition site for Trg. Our sequence also included part of the mglA gene, which is immediately distal to mglB. The amino acid sequence deduced for the beginning of the MglA protein showed homology with a family of polypeptides that contain an ATP-binding site and are components of binding-protein-dependent transport systems.     

Effects of lactation and removal of pups on the rate of triacyglycerol/fatty acid substrate cycling in white adipose tissue of the rat.

The rate of the triacylglycerol/fatty acid substrate cycle was measured in vivo in adipose tissue of virgin and lactating rats with pups removed. The rate decreased by 70% in adipose tissue of lactating rats and increased 9-fold on removal of the pups. Similar differences in cycling rate were seen in adipose tissue incubated in vitro in the presence of isoprenaline.     

Molecular cloning, cDNA structure and deduced amino acid sequence for a type I regulatory subunit of cAMP-dependent protein kinase from human testis

A 1.5 kilobase (kb) cDNA clone containing the entire coding region for a regulatory subunit of type I cAMP-dependent protein kinase (RI) was isolated from a human testis cDNA library. The cDNA clone encodes a protein of 381 amino acids that shows 98% and 97% homology to the bovine skeletal muscle RI and rat brain RI, respectively. Northern blot analysis demonstrates two major mRNA-species (1.5 and 3.0 kb) in human testis and one mRNA-species (3.0 kb) in human T-lymphocytes.     

Cellular localization and age-dependent changes in mRNA for cyclic adenosine 3',5'-monophosphate-dependent protein kinases in rat testis

Gonadotropin activation of cyclic adenosine 3',5'-monophosphate (cAMP)-dependent protein kinases plays an important role in the regulation of testicular function. This study was undertaken to establish the expression of various subunits of cAMP-dependent protein kinases in different testicular cell types as well as during sexual maturation. RNA was extracted from cultured Sertoli cells, cultured peritubular cells, germ cells (pachytene spermatocytes, round spermatids), tumor Leydig cells, as well as whole testis from rats of various ages. Messenger RNA levels were studied by Northern analysis using available cDNA probes. The regulatory subunit (R) designated RII51 was found to be predominantly expressed in cAMP-stimulated Sertoli cells and tumor Leydig cells. Much lower levels were found in cultured peritubular cells and germ cells. A 2.9- and 3.2-kb mRNA for the RI subunit were found at about similar levels in all cell types, whereas the smaller 1.7-kb mRNA was expressed in high levels in germ cells. Also, the catalytic subunit (C) of cAMP-dependent protein kinase, designated C alpha, was expressed in all cell types; the highest mRNA levels for this subunit were found in germ cells and in tumor Leydig cells. The 1.7-kb mRNA for androgen-binding protein (ABP) was abundant in cAMP-stimulated Sertoli cells and was not present in other cell types of the testis. Furthermore, the cellular localization of the cAMP-dependent protein kinase subunits was also supported by developmental studies. The mRNA level of the RII51 3.2-kb species was relatively constant until Day 30, after which there was a tendency to decrease. A 1.6-kb message first appeared at greater ages. The mRNA for the smaller 1.7-kb species of RI, as well as the C alpha, showed a significant increase during development, supporting an enrichment of these mRNAs in germ cells. Messenger RNA levels for ABP were not detected in testis from 5- to 10-day-old rats but increased up to Day 30. After this age, mRNA for ABP revealed an age-dependent decrease, which parallels the relative increase of germ cells in the testis. In summary, these results demonstrate a clear pattern of cellular localization of the various mRNA species for subunits of the cAMP-dependent protein kinase in the rat testis.     

Aspergillus myositis in a patient with a myelodysplastic syndrome

We present the case of an elderly man who, while being treated with corticosteroids for a myelodysplastic syndrome, developed myositis of the calf due to Aspergillus fumigatus. Despite therapy with amphotericin B the myositis failed to resolve and he died. At autopsy, a localized necrotizing myositis of the right calf was found with no evidence of disseminated Aspergillus infection. Myositis in the setting of disseminated candidiasis or cryptococcosis has been previously reported. This case is unique in that it is the first reported case of localized fungal myositis and of myositis caused by Aspergillus.     

Growth of Nitrobacter by dissimilatoric nitrate reduction

Abstract Eight strains of the genus Nitrobacter grew under anaerobic conditions in the presence of nitrate. The growth was inhibited by nitrate concentrations above 0.5 mM. By a special culture technique inhibition caused by nitrite was abolished. Nitrate oxidizing cells grew in gas tight culture flasks as a biofilm on a gas-permeable silicone tubing. The biofilm allowed nitrate-reducing cells to grow at a low nitrite concentration. These cells grew either actively motile in the anaerobic medium, or in anaerobic zones of the biofilm. They produced nitrite and ammonia. Nitrogen balance calculations established a loss of inorganic nitrogen for 5 of 8 strains. This implies that nitrate-reducing cells produced furthermore volatile nitrogen compounds. N₂O was detected by gas chromatography.     

Z-linked inheritance of male olfactory response to sex pheromone components in two species of tortricid moths, Ctenopseustis obliquana and Ctenopseustis sp.

The olfactory response from male pheromone sensitive sensilla was investigated in the endemic New Zealand brownheaded leafrollers Ctenopseustis obliquana (Walker) and C. sp. ropeana (Lepidoptera, Tortricidae). The responses from 281 sensilla from the parental strains and from both the reciprocal crosses, including F₁, F₂ and maternal and paternal backcrosses were recorded, and statistically analysed using a multivariate analysis.In males of both the parental strains, a large amplitude cell responded to the main pheromone component of the conspecific female, in C. obliquana (Z)-8-tetradecenyl acetate (Z8-14:OAc) and in C. sp ropeana (Z)-5-tetradecenyl acetate (Z5-14:OAc). Both male types also possessed a small amplitude cell, which in C. obliquana responded weakly to Z5-14:OAc and tetradecyl acetate (14:OAc), and in C. sp ropeana responded to Z8-14:OAc. The responses from the different types of hybrid males were more variable than the responses from parental males. A main pattern could, however be seen, corresponding with the expected pattern in a sex-linked inheritance on the Z-chromosome of a C. sp ropeana type dominant genetic factor. The more pronounced variation in the hybrids could not be explained by this model, and might be due to the involvement of additional genes.

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Résumé Les réactions olfactives des sensilles mâles sensibles aux phéromones ont été examinées par enregistrement de l'extrémité de la sensille chez les tordeuses C. obliquana Walker et C. sp. ropeana. Les enregistrements ont porté sur 281 sensilles des lignées parentales et des croisements réciproques de F₁, F₂ et de croisements en retour maternel et paternel. Les résultats des enregistrements d'une sensille ont été soumis à une analyse en composantes principales.Chez les mâles de chaque lignée parentale un seul type physiologique de sensille a été découvert; une cellule répond par un pic grand au principal constituant de la phéromone femelle conspécifique. (Z)-8-acétate tétradécényl (Z8-14:OAc) pour C. obliquana, et (Z)-5-acétate tétradécényl (Z5-14:OAc) pour C. sp. ropeana. Une seconde type de cellule dans les sensilles des deux espèces de mâles présente un pic petit pour Z5-14:OAc et pour l'acétate tétradécyl (14:OAc) chez C. obliquana, et pour C. sp. ropeana au Z8-14:OAc. Les réponses des sensilles des différents types de mâles hybrides sont plus hétérogènes que celles des sensilles de leurs pères. Un schéma général pourrait cependant être décelé, correspondant au schéma prévu avec une hérédité d'un facteur dominant liée au sexe sur le chromosome Z de C. sp. ropeana. La variation plus accentuée chez les hybrides ne peut être expliquée par ce modèle, et pourrait impliquer des gènes additionnels.

     

Human testis cDNA for the regulatory subunit RII alpha of cAMP-dependent protein kinase encodes an alternate amino-terminal region

Phosphorylations catalyzed by cAMP-dependent protein kinase are essential for sperm motility, and type II cAMP-dependent protein kinase in mature sperm has been shown to be firmly bound to the flagellum via the regulatory subunit, RII. The present study documents high-levelled expression of a human, testis-specific RII alpha mRNA (2.0 kb) analogous to the rat mRNA which is induced in haploid germ cells [(1988) FEBS Lett. 229, 391-394]. We report the molecular cloning of a full-length human cDNA corresponding to this unique testis mRNA, and the presence of an alternative amino-terminal region (amino acids 45-75) of the predicted RII alpha protein (404 amino acids) compared with the previously published mouse and rat sequences. However, this alternate region is also shown to be present in RII alpha mRNA (7.0 kb) of human somatic cells. Our data indicate the divergent amino-terminal sequence to be due to species differences, suggesting an active evolutionary pressure on this particular region, which could be involved in subcellular attachment of RII alpha and thereby localization of kinase activity to certain targets within the cell.     

Fractionation of immunoglobulins by liquid-liquid partition chromatography in aqueous two-phase systems

In this paper we show that although immunoglobulins are easily precipitated in solutions containing polyethylene glycol (PEG), especially at pH's where the conformation of the proteins should be close to native, human and rabbit IgG can be solubilized in aqueous dextran/PEG two-phase systems containing glycine and sodium chloride at pH 7.0 and that human IgA and IgM can be solubilized in such systems if the pH is increased to 9.0. Liquid-liquid partition chromatography (LLPC) on Li-ParGel was used to separate immunoglobulins into subfractions. Human IgG, IgM, and IgA all gave three peaks in the system used. These results indicate the possibility of separating different classes of immunoglobulins with this method. Specific IgG antibodies isolated from a rabbit antiserum against human serum proteins gave only two peaks in the LLPC system while the total IgG population gave three, as did human IgG. Thus, partitioning of immunoglobulins seems to be related to antibody activity.