Anti-myosin heavy chain monoclonal antibodies reveal two IIB (fast) fiber subtypes

Indirect immunofluorescence analysis of different rat skeletal muscles using anti-myosin heavy chain (MHC) monoclonal antibodies (MAb) revealed the presence of two immunologically distinct kinds of fibers within the IIB fibers, histochemically identified by myosin ATPase staining. Some IIB fibers (designated here as IIB1) were unreactive with one anti-fast MHC MAb, whereas they did react with another anti-fast MHC MAb; other IIB fibers (designated here as IIB2) reacted with both anti-fast MAbs. Neither of the two IIB fiber subtypes was significantly reactive with a neonatal MHC MAb. The number of each IIB fiber subtype was age-dependent, at least in the plantaris muscle. IIB1 fibers were observed only in the superficial portion of the plantaris and gastrocnemius muscle. The ratio of IIB1:IIB2 fibers was about the same throughout the extensor digitorum longus and extraocular muscles. Therefore, the two kinds of IIB fibers here observed have a different myosin heavy chain content. On the basis of their specific immunoreactivities, we suggest that IIB1 fibers contain the previously described MHCB. IIB2 fibers contain either a unique new MHC isoform or a mixture of at least two MHC, possibly composed of the MHCB and either the previously described MHCA or a new MHC isoform.     

Growth of Phytomonas serpens in a Defined Medium; Nutritional Requirements

ABSTRACT. Three strains of Phytomonas serpens two from tomatoes, Lycopersicon esculentum one from the insect Phtia picta (Hemiptera, Coreidae), were cultivated in a chemically defined medium developed from a defined medium for cultivating insect flagellates. Besides organic growth factors required by other insect trypanosomatids this flagellate requires, serine and inositol. Glutamine stimulates growth, and, surprisingly, does not require heme.     

Effect of oxygen transfer and carbon source composition on the growth of Corynebacterium melassecola

Corynebacterium melassecola, a glutamate producing bacteria, has been cultivated in a 4-l batch aerated fermentor with glucose and fructose mixtures of various proportions as carbon sources and mineral ammonium as nitrogen source. Experimental results are analysed in order to obtain the specific rates and the conversion yields. A data reconciliation technique is used to manage with the experiments performed in similar conditions. The redundant experimental informations are used to test the carbon, nitrogen, available electrons, phosphorus and protons balances. The measurement of dissolved oxygen tension and volumetric oxygen transfer coefficient indicates that oxygen was never a limiting substrate in the experimental device though the oxygen demand was up to 70 mmoles/l.h and the heterogeneity of the liquid phase. The highest specific growth rate (0.59 h^?1) and the highest biomass production yield (0.48 gX/gS) are obtained with glucose as the sole carbon source. These two parameters progressively decrease with the increase of the proportion of fructose in the medium down to 0.43 h^?1 and 0.38 gX/gS on fructose so that the specific carbon uptake rate remains constant and equal to 0.04 C-mole/gX h. From a metabolic point of view, this decrease of the carbon conversion efficiency into cellular material is correlated to a lower carbon flux through the pentose pathway as the fructose proportion is increased. This is correlated with an accumulation of fructose 1,6-biphosphate which further activates the lactate deshydrogenase resulting to lactate production though the metabolism remains strictly aerobic.     

Sequence variation in the outer-surface-protein genes of Borrelia burgdorferi

Borrelia burgdorferi is a spirochete pathogen transmitted among warm- blooded hosts by ixodid ticks. Frequency-dependent selection for variant outer-surface proteins might be expected to arise in this species, since rare variants are more likely to avoid immune surveillance in previously infected hosts. We sequenced the OspA and OspB genes of nine North American strains and compared them with nine strains previously described. For each gene, the mean number of synonymous substitutions per synonymous site and the mean number of nonsynonymous substitutions per nonsynonymous site show only a twofold excess of silent mutations. Synonymous rates vary widely along the OspB protein. Some regions show a significant excess of silent substitutions, while divergence in other regions is constrained by biased base composition or selection. The presence, in antigenically important regions of the protein, of significant variation among strains, as well as evidence for recombination among strains, should be considered in attempts to develop vaccines against this disease.      

Characterisation of a 34 kD protein from potato cyst nematodes, using monoclonal antibodies with potential for species diagnosis

Two monoclonal antibodies, which differentially recognise the two species of potato cyst nematodes (PCN), Globodera pallida and G. rostochiensis, are described. They have been shown to have potential for quantification of these two species, recognising proteins of the same molecular weight (34 kD) in both species. Further investigation showed these proteins to have isoelectric points at pH values of 5.7 in G. pallida and 5.9 in G. rostochiensis, in common with the proteins used by Fleming & Marks (1983) to differentiate the species of PCN. They are likely to be structurally very similar, with the same physiological function (and therefore similar concentrations) in the two species. In cross-reactivity tests with a wide range of soil nematode species, the antibodies reacted strongly only with species of the genus Globodera, and thereby confirmed their potential as the basis of a quantitative immunoassay likely to be useful in management of PCN populations.     

The Role of Maize in the Epidemiology of Barley Yellow Dwarf Virus in Northeast Spain

Barley yellow dwarf virus has been detected in maize by indirect enzyme-linked immunosorbent assay (ELISA) and by immunospecific electron microscopy (ISEM). Samples of maize collected in September 1988, 1989 and 1990 showed that this crop is an important reservoir of BYDV; MAV-like isolates were the most common although PAV-like and RPV-like isolates were also present. Earlier research in Spain had shown that PAV-like isolates were predominant. Thus the evidence from this work that MAV was the main isolate, and very widely spread, is important for future research on BYDV epidemiology in Spain.     

The structure and function of p55PIK reveal a new regulatory subunit for phosphatidylinositol 3-kinase.

Phosphatidylinositol 3-kinase (PI-3 kinase) is implicated in the regulation of diverse cellular processes, including insulin-stimulated glucose transport. PI-3 kinase is composed of a 110-kDa catalytic subunit and an 85-kDa regulatory subunit. Here, we describe p55PIK, a new regulatory subunit that was isolated by screening expression libraries with tyrosine-phosphorylated insulin receptor substrate 1 (IRS-1). p55PIK is composed of a unique 30-residue NH2 terminus followed by a proline-rich motif and two Src homology 2 (SH2) domains with significant sequence identify to those in p85. p55PIK mRNA is expressed early during development, remains abundant in adult mouse brain and testis tissue, and is detectable in adult adipocytes and heart and kidney tissues. p55PIK forms a stable complex with p110, and it associates with IRS-1 during insulin stimulation. Moreover, the activated insulin receptor phosphorylates p55PIK in Sf9 cells, and insulin stimulates p55PIK phosphorylation in CHOIR/p55PIK cells. The unique features of p55PIK suggest that it is important in receptor signaling.