Oligomeric properties and signal peptide binding by Escherichia coli Tat protein transport complexes

The Escherichia coli Tat apparatus is a protein translocation system that serves to export folded proteins across the inner membrane. The integral membrane proteins TatA, TatB and TatC are essential components of this pathway. Substrate proteins are directed to the Tat apparatus by specialized N-terminal signal peptides bearing a consensus twin-arginine sequence motif. Here we have systematically examined the Tat complexes that can be purified from overproducing strains. Our data suggest that the TatA, TatB and TatC proteins are found in at least two major types of high molecular mass complex in detergent solution, one consisting predominantly of TatA but with a small quantity of TatB, and the other based on a TatBC unit but also containing some TatA protein. The latter complex is shown to be capable of binding a Tat signal peptide. Using an alternative purification strategy we show that it is possible to isolate a TatABC complex containing a high molar excess of the TatA component.     

DNA analyses reveal abundant homoplasy in taxonomically important morphological characters of Eusiroidea (Crustacea,Amphipoda)

Eusiroidea is one of the 20 amphipod superfamilies that were erected to subdivide the very large and controversial suborder Gammaridea. Yet, the definition of the superfamily is not based on synapomorphies, but on a combination of diagnostic phenetic similarities that hold more or less consistently across families. Moreover, many of the characters used to define eusiroid families are suspected to show convergent evolution. The current classification of the Eusiroidea may therefore not reflect evolutionary relationships accurately. Here, we present a molecular phylogenetic re‐analysis of the Eusiroidea based on a comparison of 18S and 28S rDNA sequences of 73 species, representing 47 genera and 16 families that potentially belong to the superfamily. The results suggest that at least species belonging to 14 of these traditional families would be part of a eusiroid clade, increasing by more than twofold the species and generic richness of the group. However, most of the eusiroid families surveyed do not appear monophyletic. Finally, the analyses show that several important morphological characteristics, traditionally used in eusiroid taxonomy, are homoplastic.     

Diurnal variations in ventilatory and cardiorespiratory responses to submaximal treadmill exercise in females

Diurnal variations in ventilatory and cardiorespiratory responses to submaximal treadmill exercise were analysed in 11 eumenorrhoeic women and in 10 women using monophasic oral contraceptives. Subjects performed submaximal treadmill exercise at three intensities averaging 7, 8, and 9 km x h(-1), each for 4 min at 0800, 1300 and 1700 hours, assigned randomly on 3 separate days. Rectal temperature was measured before (T(rec(b))) and after (T(rec(a))) exercise. Cardiac frequency (f(c)), ventilation (V(E)), oxygen uptake (VO(2)), carbon dioxide output (VCO(2)), and respiratory exchange ratio (R) were assessed in the last minute of each stage of the exercise. Both T(rec(b)) and T(rec(a)) increased from 0800 to 1700 hours (P < 0.001). For a given submaximal work rate, VO(2) and VCO(2) were higher in the afternoon compared to the morning. Similarly, R was increased at 1700 hours compared to 0800 hours during the recovery period following exercise (P < 0.05). However, V(E) did not vary significantly during the day at any of the running intensities. No significant interactions (group x time of day) were observed in any of the studied parameters. In contrast to ventilation, the VO(2) and VCO(2) of the females during submaximal exercise were both affected by the time of day, without any differences between eumenorrhoeic women and users of oral contraceptives.     

Exosomes in human semen carry a distinctive repertoire of small non-coding RNAs with potential regulatory functions

Semen contains relatively ill-defined regulatory components that likely aid fertilization, but which could also interfere with defense against infection. Each ejaculate contains trillions of exosomes, membrane-enclosed subcellular microvesicles, which have immunosuppressive effects on cells important in the genital mucosa. Exosomes in general are believed to mediate inter-cellular communication, possibly by transferring small RNA molecules. We found that seminal exosome (SE) preparations contain a substantial amount of RNA from 20 to 100 nucleotides (nts) in length. We sequenced 20–40 and 40–100 nt fractions of SE RNA separately from six semen donors. We found various classes of small non-coding RNA, including microRNA (21.7% of the RNA in the 20–40 nt fraction) as well as abundant Y RNAs and tRNAs present in both fractions. Specific RNAs were consistently present in all donors. For example, 10 (of ∼2600 known) microRNAs constituted over 40% of mature microRNA in SE. Additionally, tRNA fragments were strongly enriched for 5’-ends of 18–19 or 30–34 nts in length; such tRNA fragments repress translation. Thus, SE could potentially deliver regulatory signals to the recipient mucosa via transfer of small RNA molecules.     

An information-theoretic analysis of genetics, gender and age in cancer patients

Germline genetics, gender and hormonal-signaling pathways are all well described modifiers of cancer risk and progression. Although an improved understanding of how germline genetic variants interact with other cancer risk factors may allow better prevention and treatment of human cancer, measuring and quantifying these interactions is challenging. In other areas of research, Information Theory has been used to quantitatively describe similar multivariate interactions. We implemented a novel information-theoretic analysis to measure the joint effect of a high frequency germline genetic variant of the p53 tumor suppressor pathway (MDM2 SNP309 T/G) and gender on clinical cancer phenotypes. This analysis quantitatively describes synergistic interactions among gender, the MDM2 SNP309 locus, and the age of onset of tumorigenesis in p53 mutation carriers. These results offer a molecular and genetic basis for the observed sexual dimorphism of cancer risk in p53 mutation carriers and a model is proposed that suggests a novel cancer prevention strategy for p53 mutation carriers.     

Comparative post-hatching ontogeny of Pseudoplatystoma reticulatum (Eigenmann & Eigenmann 1889) and Leiarius marmoratus (Gill, 1870) and their hybrid

Ontogenetic studies of the eggs and larvae of fish can provide information on the initial life history and biology of a species, are important for taxonomic and evolutionary studies, and for cultivation in captivity. The aim of this study was to analyze and describe the main morphological differences in the larval ontogeny of Pseudoplatystoma reticulatum, Leiarius marmoratus, and its hybrid (♀ P. reticulatum × ♂ L. marmoratus), as well as to identify characteristics that can identify the species and their hybrid at the larvae and juvenile stages. 205 L. marmoratus, 210 P. reticulatum, and 205 hybrid specimens were analyzed, all of which were obtained through induced reproduction. Analyses were performed from hatching to 30 days post-hatching. 19 morphometric and 5 meristic characteristics were evaluated, in addition to chromatophore shape and distribution. The specimens were classified into two life periods: Larval (stages: yolk sac, pre-flexion, flexion, and post-flexion) and Juvenile. Newly hatched larvae were transparent, poorly developed, and had a scarcity of chromatophores. During the early stages of larval development, the three groups showed similarities in appearance and proportional dimensions. However, at both the end of the post-flexion stage and at the juvenile period when individuals were approximately 2 cm long, it was possible to differentiate between hybrids and their parental species by their morphometric, meristic, and pigment characteristics. The hybrid, despite occupying an intermediate position in relationship to its parents, exhibited a shape and size more similar to P. reticulatum, its maternal parent.     

Irreversible inhibition of aldolase by a phosphorylated alpha-dicarbonyl compound

The preparation of a phosphorylated alpha-dicarbonyl compound designed to specifically react with arginine residues of enzymes accepting phosphorylated compounds as effectors is reported, and shown to inhibit rabbit muscle aldolase in a time-dependent and irreversible manner. This irreversible inhibition occured in a buffer devoid of borate ions, suggesting that the presence of the phosphate moiety contributes in the stabilization of the adduct formed with arginine residues. Under the same conditions, the metalloenzyme iron superoxide dismutase, in which an arginine is known to be critical for the catalytic function, is not significantly inhibited.     

Biological aspects of chronic lymphoid leukemia

The present study is concerned with clinical prognostic classifications of chronic lymphocytic leukemia derived from numerous statistics and multivariate survival analysis. We analyse the prognostic significance of some biological patterns: bone marrow histological studies, lymph node biopsies, size, structure and doubling time of blood lymphocytes, tumoral markers and karyotype analysis. The problem of the monoclonal clone origin is discussed: differentiation abnormality or proliferation of a physiological clone.