The toad fly Lucilia bufonivora: its evolutionary status and molecular identification

The blow fly genus Lucilia is composed largely of saprophages and facultative myasis agents, including the economically important species Lucilia cuprina (Wiedemann) (Diptera: Calliphoridae) and Lucilia sericata (Meigen). Only one species is generally recognized as an obligate agent of myiasis, Lucilia bufonivora Moniez, and this is an obligate parasite of toads. Lucilia silvarum (Meigen), a sister species, behaves mainly as a carrion breeder; however, it has also been reported as a facultative parasite of amphibians. Morphologically, these species are almost identical, and historically this has led to misidentification, taxonomic ambiguity and a paucity of studies of L. bufonivora. In this study, dipterous larvae were analysed from toad myiasis cases from the U.K., The Netherlands and Switzerland, together with adult specimens of fly species implicated in amphibian parasitism: L. bufonivora, L. silvarum and Lucilia elongata Shannon (from North America). Partial sequences of two genes, cox1 and ef1α, were amplified. Seven additional blow fly species were analysed as outgroups. Bayesian inference trees of cox1, ef1α and a combined‐gene dataset were constructed. All larvae isolated from toads were identified as L. bufonivora and no specimens of L. silvarum were implicated in amphibian myiasis. This study confirms L. silvarum and L. bufonivora as distinct sister species and provides unambiguous molecular identification of L. bufonivora.     

PAK-family kinases regulate cell and actin polarization throughout the cell cycle of Saccharomyces cerevisiae

During the cell cycle of the yeast Saccharomyces cerevisiae, the actin cytoskeleton and cell surface growth are polarized, mediating bud emergence, bud growth, and cytokinesis. We have determined whether p21-activated kinase (PAK)-family kinases regulate cell and actin polarization at one or several points during the yeast cell cycle. Inactivation of the PAK homologues Ste20 and Cla4 at various points in the cell cycle resulted in loss of cell and actin cytoskeletal polarity, but not in depolymerization of F-actin. Loss of PAK function in G1 depolarized the cortical actin cytoskeleton and blocked bud emergence, but allowed isotropic growth and led to defects in septin assembly, indicating that PAKs are effectors of the Rho-guanosine triphosphatase Cdc42. PAK inactivation in S/G2 resulted in depolarized growth of the mother and bud and a loss of actin polarity. Loss of PAK function in mitosis caused a defect in cytokinesis and a failure to polarize the cortical actin cytoskeleton to the mother-bud neck. Cla4-green fluorescent protein localized to sites where the cortical actin cytoskeleton and cell surface growth are polarized, independently of an intact actin cytoskeleton. Thus, PAK family kinases are primary regulators of cell and actin cytoskeletal polarity throughout most or all of the yeast cell cycle. PAK-family kinases in higher organisms may have similar functions.     

Chemical modification of the actin binding site of rabbit muscle aldolase by diethylpyrocarbonate

Summary To extend the available information on the significance of the interactions between glycolytic enzymes and the actin component of the cellular ultrastructure, investigations into the compositional characteristics of the actin binding site on one of the major glycolytic enzymes, aldolase, have been undertaken. As the electrostatic nature of the association has been previously reported indicative of a cationic region on the enzyme involved in the binding, these studies have investigated the possibility of the involvement of histidine residues in this binding region. By the use of the histidine specific reagent, diethylpyrocarbonate, we have been able to establish a difference in nature of an actin binding domain and the active site domain which does contain an essential histidine. The results have been discussed in relation to the significance of this finding with respect to the binding of aldolase to subcellular structure.     

Prenatal onset of Axonopathy in Dystonia musculorum mice

Dystonia musculorum (dt) is a recessive hereditary neuropathy of the mouse. Affected animals display loss of limb coordination and twisting of the trunk. Sensory nerve fibers of these mice are severely reduced in number, and the remaining fibers present numerous axonal swellings. The gene defective in dt, dystonin (Dst), encodes a cytoskeletal linker protein that forms the bridge between F-actin and intermediate filaments. Dst is expressed during embryogenesis, whereas overt phenotype in dt mice only appears during the second week after birth. Here we show that axonal swellings are present in sensory nerve fibers of dt embryos as early as E15.5, before myelination and radial axonal growth have begun. Thus disease progression is gradual in dt mice, having begun during embryogenesis. In dt embryos, microtubule network disorganization and cytoplasmic organelle accumulation within axonal swellings were consistently observed. In addition, a few of the axonal swellings presented intermediate filament accumulation. These results demonstrate that dystonin is required for cytoskeleton organization during axonogenesis. They also suggest that axonal transport defects, through microtubule network perturbation, may be the primary mechanism of neurodegeneration in dt mice. Dev. Genet. 22:160–168, 1998. © 1998 Wiley-Liss, Inc.     

Mammalian ER stress sensor IRE1β specifically down-regulates the synthesis of secretory pathway proteins

Accumulation of unfolded proteins in the endoplasmic reticulum (ER) causes ER stress. The ER stress sensor inositol requiring enzyme-1beta (IRE1β), which is specifically expressed in intestinal epithelial cells, is thought to be involved in translational repression. However, its mechanism of action is not fully understood. Using a reporter that can evaluate and distinguish between translation efficiency in the cytosol and on the ER membrane, we show here that IRE1β represses translation on the ER membrane but not in the cytosol, and that this selective repression depends on the RNase activity of IRE1β.     

Class VI myosin moves processively along actin filaments backward with large steps.

Among a superfamily of myosin, class VI myosin moves actin filaments backwards. Here we show that myosin VI moves processively on actin filaments backwards with large ( approximately 36 nm) steps, nevertheless it has an extremely short neck domain. Myosin V also moves processively with large ( approximately 36 nm) steps and it is believed that myosin V strides along the actin helical repeat with its elongated neck domain that is critical for its processive movement with large steps. Myosin VI having a short neck cannot take this scenario. We found by electron microscopy that myosin VI cooperatively binds to an actin filament at approximately 36 nm intervals in the presence of ATP, raising a hypothesis that the binding of myosin VI evokes "hot spots" on actin filaments that attract myosin heads. Myosin VI may step on these "hot spots" on actin filaments in every helical pitch, thus producing processive movement with 36 nm steps.     

盐酸氨基葡萄糖胶囊对膝骨关节炎患者血清IL-1β和TNF-α水平的影响及临床疗效

目的:观察盐酸氨基葡萄糖对膝骨关节炎(Knee osteoarthritis,KOA)患者血清IL-1β和TNF-α水平的影响及临床治疗效果。方法:将128例KOA患者随机分为治疗组与对照组,治疗组口服盐酸氨基葡萄糖胶囊,对照组口服双氯芬酸钠缓释胶囊,各组均以8周为一个疗程。用流式细胞仪微球阵列法检测两组患者治疗前后血清IL-1β和TNF-α水平,比较两组治疗前后膝关节功能WOMAC评分、疼痛VAS评分的变化及疼痛缓解时间。结果:治疗后,两组患者血清IL-1β和TNF-α的水平均较治疗前明显降低(P0.05),WOMAC评分较治疗前显著升高(P0.05),且治疗组血清IL-1β和TNF-α的水平均显著低于对照组(P0.05),而WOMAC评分明显高于对照组(P0.05)。两组患者疼痛VAS评分和疼痛缓解时间比较差异无统计学意义(P0.05)。结论:盐酸氨基葡萄糖胶囊能明显改善KOA患者膝关节功能,缓解膝关节局部的疼痛,可能与其显著降低患者血清IL-1β和TNF-α的水平有关。