Morphology and molecular phylogeny of Pseudotrichonympha hertwigi and Pseudotrichonympha paulistana (Trichonymphea, parabasalia) from neotropical rhinotermitids

Pseudotrichonympha is a large hypermastigote parabasalian found in the hindgut of several species of rhinotermitid termites. The genus was discovered more than 100 years ago, and although over a dozen species have since been described, this represents only a small fraction of its likely diversity: the termite genera from which Pseudotrichonympha is known are all species rich, and in most cases their hindgut symbionts have not been examined. Even formally described species are mostly lacking in detailed microscopic data and/or sequence data. Using small subunit ribosomal RNA gene sequences and light and scanning electron microscopy we describe here the morphology and molecular phylogenetic position of two Pseudotrichonympha species: the type species for the genus, Pseudotrichonympha hertwigi from Coptotermes testaceus (described previously in line drawing only), and Pseudotrichonympha paulistana from Heterotermes tenuis (described previously based on light microscopy only).     

Wound-induced vascular occlusions in tissues of the reed Phragmites australis: their development and chemical nature

This work focuses on the development of vascular occlusions, which are gels resealing the wounded vascular systems of injured organs, in the common reed Phragmites australis. Their formation seems to be crucial in keeping the internal environment of the plant stable. Histochemical tests, combined with an extraction series, were used to follow changes in the chemical nature of gels during their development. It was found that the first gel material was secreted by living cells in the vicinity of the incision within 1 or 2 d after wounding. Early gels were colourless and mainly composed of acidic polysaccharides interlinked by Ca2+ bridges. The properties of the gel material gradually changed during maturation. The matrix of polysaccharides in the early gels was later modified and interlinked by other components, resulting in a highly resistant material. Structural proteins were identified as the principal interlocking components of the material, and were responsible for its high resistance.     

Chromatographic detection of residual cellular DNA on short monolithic columns

Analysis of crude samples from biotechnological processes is often required to demonstrate that residual host cell impurities are reduced or eliminated during purification. Current knowledge suggests that a continuous-cell-line DNA can be considered a cellular contaminant rather than a significant risk factor requiring removal to extremely low levels. Anion-exchange chromatography is one of the most important methods used in the downstream processing and analysis of different biomolecules. In this article, an application using Convective Interaction Media monolithic columns to improve the detection of residual cellular DNA is described.     

Molecular cytogenetic analysis of recently evolved Tragopogon (Asteraceae) allopolyploids reveal a karyotype that is additive of the diploid progenitors

Tragopogon mirus and T. miscellus (both 2n = 4x = 24) are recent allotetraploids derived from T. dubius × T. porrifolius and T. dubius × T. pratensis (each 2n = 2x = 12), respectively. The genome sizes of T. mirus are additive of those of its diploid parents, but at least some populations of T. miscellus have undergone genome downsizing. To survey for genomic rearrangements in the allopolyploids, four repetitive sequences were physically mapped. TPRMBO (unit size 160 base pairs [bp]) and TGP7 (532 bp) are tandemly organized satellite sequences isolated from T. pratensis and T. porrifolius, respectively. Fluorescent in situ hybridization to the diploids showed that TPRMBO is a predominantly centromeric repeat on all 12 chromosomes, while TGP7 is a subtelomeric sequence on most chromosome arms. The distribution of tandem repetitive DNA loci (TPRMBO, TGP7, 18S-5.8S-26S rDNA, and 5S rDNA) gave unique molecular karyotypes for the three diploid species, permitting the identification of the parental chromosomes in the polyploids. The location and number of these loci were inherited without apparent changes in the allotetraploids. There was no evidence for major genomic rearrangements in Tragopogon allopolyploids that have arisen multiple times in North America within the last 80 yr.     

Gender-related differences in maximum mechanical power output in short-term activities in children and adolescents

The study was designed to examine the gender-related differences in maximum mechanical power output in various short-burst activities during growth. The subject sample consisted of four subgroups: 9 boys (14.11 +/- 0.6 yr), 9 boys (10.67 +/- 0.71 yr), 7 girls (14.29 +/- 0.49 yr), 7 girls (10.57 +/- 0.54 yr). We measured peak power (PP), mean power (MP), fatigue index (FI) during 30-s WAnT, squat jump height (SJH) and power (SJP), and counter movement jump height (CMJH) and power (CMJP), maximum speed over 20-metre distance (S20). Lactation concentration was measured in the 3rd and 5th minutes after the WAnT Ratio normalisation and ANCOVA were used to remove the influence of the differences in muscle (MM) and body mass (BM). Male adolescents had higher absolute values of PP (P < 0.05), MP (P < 0.05) than female. Ratio normalisation showed that boys had higher PP/BM (P < 0.05), PP/MM (P < 0.05), MP/BM (P < 0.05), MP/MM (P < 0.06) than girls. The ANCOVA adjustment for MM showed differences between genders in PP (P < 0.001), MP (P < 0.001), SJH (P < 0.05), SJP (P < 0.05) and CMJP (P < 0.001), whereas the ANCOVA adjustment for BM showed differences only in PP (P < 0.001), MP (P < 0.001). Prepubertal boys had higher absolute values only in SJP (P < 0.05). We concluded that variations in body composition could not be the only key to gender-related differences in power output in short-burst activities.     

Invasiveness of transformed human breast epithelial cell lines is related to cathepsin B and inhibited by cysteine proteinase inhibitors

The activities'of the lysosomal cysteine proteinases cathepsin B and L are regulated by their endogenous inhibitors, stefins A and B, and cystatin C, and their imbalance may be associated with increased invasiveness and development of the malignant cell phenotype. The aim of this study was to investigate mRNA, protein and activity levels of the above proteins in relation to in vitro invasiveness and to the reported in vivo tumorigenicity of four human breast tumor cell lines: the spontaneously immortalized cell line MCF10A, its c-Ha-ras transfectant MCF10AT, and two tumorigenic derivative cell lines, MCF10AT-Ca1a and MCF10AT-Ca1d. Invasiveness did not correlate with tumorigenicity, since the MCF10AT cell was the most invasive and the remaining three were at about half of its level. Cathepsin B expression paralleled the in vitro invasiveness through matrigel at all levels of expression, but cathepsin L did not. Stefin levels were elevated several-fold in the tumorigenic cell lines, but not in MCF10AT. The hypothesis that cathepsin B plays an active role in the invasion of breast cancer cell lines was confirmed by the fact that synthetic cysteine proteinase inhibitors, particularly those selective for cathepsin B, significantly reduced the invasion of the MCF10AT cells.     

Okadaic acid induces sustained activation of NFkappaB and degradation of the nuclear IkappaBalpha in human neutrophils

Human neutrophils differ from other cells by containing high amount of IkappaBalpha in the nucleus, and this increased nuclear IkappaBalpha accumulation is associated with the inhibition of NFkappaB activity and increased apoptosis. However, the mechanisms regulating NFkappaB activation and IkappaBalpha degradation in human neutrophils are little understood. The objective of this study was to provide a further insight into the mechanisms regulating NFkappaB activity and IkappaBalpha degradation in human neutrophils. We show that okadaic acid (OA), an inhibitor of protein phosphatases PP1 and PP2A, induces sustained activation of NFkappaB and degradation of the nuclear IkappaBalpha, and increases interleukin-8 expression in the neutrophils. Furthermore, inhibitors of protein kinase C-delta (PKCdelta) and IkappaB kinase (IKK) inhibit the OA-induced activation of NFkappaB. Collectively, our results indicate that in human neutrophils, the sustained activation of NFkappaB is regulated by a continuous phosphorylation and degradation of the nuclear IkappaBalpha.