The expanding spectrum of nuclear gene mutations in mitochondrial disorders

Our understanding of the molecular basis of mitochondrial disorders has come primarily from the discovery of an expanding number of mutations of mtDNA. However, a variety of recent observations indicate that many syndromes are due to abnormalities in nuclear genes related to oxidative-phosphorylation (OXPHOS). Nuclear genes encode hundreds of proteins involved in mitochondrial OXPHOS. Nevertheless, the identification of these genes has proceeded at a much slower pace, compared with the discovery and characterization of mtDNA mutations. This scenario is rapidly changing, thanks to the discovery of several OXPHOS-related human genes, and to the identification of mutations responsible for different clinical syndromes.     

Cloning of dynein intermediate chain cDNA of the newt Cynops pyrrhogaster.

To isolate genes whose expression is up-regulated after initiation of meiosis, we employed an mRNA differential display method using RNA extracted from newt testis fragments in the spermatogonial and spermatocyte stages. We report here isolation of a spermatocyte stage-specific cDNA clone encoding a newt homologue of dynein intermediate chain (IC). The newt dynein IC cDNA was found to encode a polypeptide consisting of 694 amino acid residues with 66.8% and 45.8% amino acid sequence similarity to sea urchin dynein IC3 and Chlamydomonas IC69, respectively. The predicted protein contains five WD repeats and a novel repeated motif in the C-terminal region. Northern blot analysis revealed that newt dynein IC mRNA was expressed in the spermatocyte and round spermatid stages, suggesting that dynein IC plays a role in formation of flagella as well as in meiotic events.     

Different respiratory syncytial virus and Quillaja saponin formulations induce murine peritoneal cells to express different proinflammatory cytokine profiles

The recognition of a pathogen or a vaccine antigen formulation by cells in the innate immune system leads to production of proinflammatory cytokines, which will determine the ensuing acquired immune response quantitatively and qualitatively. Tumour necrosis factor (TNF)-alpha, interleukin (IL)-1 and IL-6 are the first set of cytokines produced upon such an encounter, which have roles both in protective immunity and immunopathogenesis evident with respiratory syncytial virus (RSV). RSV antigens in different physical adjuvant-vaccine formulations were analysed for their capacity to provoke cultured murine peritoneal cells to produce these three proinflammatory cytokines. RSV immunostimulating complex (ISCOM), i.e. both antigen and adjuvant are incorporated in the same particle, induced high levels of IL-1alpha being of the same magnitude or higher than those of live RSV and lipopolysaccharide (LPS). Live virus and LPS induced higher levels of IL-6 and TNF-alpha than ISCOM and so did non-adjuvanted UV-inactivated RSV but only at high doses. ISCOM-Matrix, i.e. ISCOM without antigens, admixed as a separate entity to inactivated RSV, downregulated or blocked the cytokine response to the inactivated RSV in contrast to ISCOM. Kinetic studies showed that ISCOM induced cytokine production first detected at hours 1, 2, 4 for TNF-alpha, IL-6 and IL-1alpha respectively, which was earlier than for the other antigen formulations containing corresponding doses of antigen and/or Quillaja adjuvant. Peak values for production of TNF-alpha and IL-6 were at 8 h and for IL-1alpha at 72 h following stimulation with ISCOM. The delayed appearance of IL-1alpha may reflect the cell-bound nature of this cytokine.     

A sampling algorithm for segregation analysis

Methods for detecting Quantitative Trait Loci (QTL) without markers have generally used iterative peeling algorithms for determining genotype probabilities. These algorithms have considerable shortcomings in complex pedigrees. A Monte Carlo Markov chain (MCMC) method which samples the pedigree of the whole population jointly is described. Simultaneous sampling of the pedigree was achieved by sampling descent graphs using the Metropolis-Hastings algorithm. A descent graph describes the inheritance state of each allele and provides pedigrees guaranteed to be consistent with Mendelian sampling. Sampling descent graphs overcomes most, if not all, of the limitations incurred by iterative peeling algorithms. The algorithm was able to find the QTL in most of the simulated populations. However, when the QTL was not modeled or found then its effect was ascribed to the polygenic component. No QTL were detected when they were not simulated.     

微生物合成中链聚羟基烷酸酯研究进展

某些微生物细胞在特定营养限制的条件下会产生聚羟基烷酸酯作为碳源储备。和短链聚羟基烷酸酯(PHB)一样 ,中链聚羟基烷酸酯由于具有更优良的性能、更高的附加值和更广泛的用途而受到人们的关注 ;此外 ,中链聚羟基烷酸酯还可以被人工合成为具有功能性侧链的半合成高聚物 ,并因此能够具有更好的弹性和更理想的结晶性能等优点 ,从而成为近年来对环境友好的生物可降解材料的研究重点。在能够合成中链聚羟基烷酸酯的微生物中 ,食油假单胞菌是最典型 ,也是研究得最多的一种。本文对由食油假单胞菌合成中链聚羟基烷酸酯的特点、代谢机制、发挥过程等内容进行了综述 ,并提出了这一研究领域未来可能的研究方向     

Non‐muscle myosin IIB helps mediate TNF cell death signaling independent of actomyosin contractility (AMC)

Non‐muscle myosin II (NM II) helps mediate survival and apoptosis in response to TNF‐alpha (TNF), however, NM II's mechanism of action in these processes is not fully understood. NM II isoforms are involved in a variety of cellular processes and differences in their enzyme kinetics, localization, and activation allow NM II isoforms to have distinct functions within the same cell. The present study focused on isoform specific functions of NM IIA and IIB in mediating TNF induced apoptosis. Results show that siRNA knockdown of NM IIB, but not NM IIA, impaired caspase cleavage and nuclear condensation in response to TNF. NM II's function in promoting cell death signaling appears to be independent of actomyosin contractility (AMC) since treatment of cells with blebbistatin or cytochalasin D failed to inhibit TNF induced caspase cleavage. Immunoprecipitation studies revealed associations of NM IIB with clathrin, FADD, and caspase 8 in response to TNF suggesting a role for NM IIB in TNFR1 endocytosis and the formation of the death inducing signaling complex (DISC). These findings suggest that NM IIB promotes TNF cell death signaling in a manner independent of its force generating property. J. Cell. Biochem. 9999: 1365–1375, 2010. © 2010 Wiley‐Liss, Inc.     

Eigenschaften des Elektronentransportsystem von Mitochondrien des Protisten Acanthamoeba castellanii Neff. I. Zum Einfluß von Hemmstoffen auf verschiedene Akzeptorsysteme, getested mit einer modifizierten Thunbergtechnik

ZUSAMMENFASSUNG. Die Reduktionsgeschwindigkeit künstlicher Elektronenakzeptoren wurde mittels einer modifizierten Thunbergtechnik in Gegenwart isolierter Mitochondrien des Protisten Acanthamoeba castellanii Neff photometrisch gemessen. Die mit verschiedenen Elektronenakzeptoren und Atmungsketteninhibitoren gewonnenen Meßergebnisse erlauben uns folgendes Bild von der Konstitution der Atmungskette zu entwerfen: a) Der Elektronentransport läuft mindestens bis zum Cytochrom b /Coenzym Q-Komplex auf zwei verschiedenen Wegen ab. b) Eine Stimulierung sowohl des Succinat-Jodnitrotetrazolium-chlorid als auch des NADH-Ferricyanid Reduktasekomplexes unter dem Einfluß von Antimycin A läßt vermuten, daß in der Atmungskette dieses Protisten gewisse Nebengleise des Elektronentransports besonders gangbar sind.
SYNOPSIS. The reduction of artificial electron acceptors by isolated mitochondria of Acanthamoeba castellanii was measured by a modified Thunberg technic. The results with different electron acceptors and respiratory chain inhibitors suggest the following scheme for the constitution of the respiratory chain: a) the chain is divided into 2 different sequences, at least up to the cytochrome b /coenzyme Q complex. b) As seen from the stimulation of the succinate-iodonitrotetrazolium chloride and NADH-ferricyanide reductase complexes by antimycin A, certain alternate pathways of electron transport become more important than the normal one.