Quantitation of fibrinolytic agents released in tissue culture

Zusammenfassung Es wird eine neue Methode zur Bestimmung der freigesetzten Menge fibrinolytisch wirksamer Stoffe in Gewebekulturen beschrieben. Das Gewebe wird in Leightonröhrchen mit einem standardisierten Fibringerinnsel gezüchtet. Die abgebaute Fibrinmenge — als Mass der fibrinolytischen Aktivität — wird durch immunologische Bestimmung der Spaltprodukte gemessen.     

Chronic lymphocytic leukaemia is driven by antigen-independent cell-autonomous signalling

B-cell antigen receptor (BCR) expression is an important feature of chronic lymphocytic leukaemia (CLL), one of the most prevalent B-cell neoplasias in Western countries. The presence of stereotyped and quasi-identical BCRs in different CLL patients suggests that recognition of specific antigens might drive CLL pathogenesis. Here we show that, in contrast to other B-cell neoplasias, CLL-derived BCRs induce antigen-independent cell-autonomous signalling, which is dependent on the heavy-chain complementarity-determining region (HCDR3) and an internal epitope of the BCR. Indeed, transferring the HCDR3 of a CLL-derived BCR provides autonomous signalling capacity to a non-autonomously active BCR, whereas mutations in the internal epitope abolish this capacity. Because BCR expression was required for the binding of secreted CLL-derived BCRs to target cells, and mutations in the internal epitope reduced this binding, our results indicate a new model for CLL pathogenesis, with cell-autonomous antigen-independent signalling as a crucial pathogenic mechanism.     

Structural basis for broad DNA-specificity in integron recombination

Lateral DNA transfer--the movement of genetic traits between bacteria--has a profound impact on genomic evolution and speciation. The efficiency with which bacteria incorporate genetic information reflects their capacity to adapt to changing environmental conditions. Integron integrases are proteins that mediate site-specific DNA recombination between a proximal primary site (attI) and a secondary target site (attC) found within mobile gene cassettes encoding resistance or virulence factors. The lack of sequence conservation among attC sites has led to the hypothesis that a sequence-independent structural recognition determinant must exist within attC. Here we report the crystal structure of an integron integrase bound to an attC substrate. The structure shows that DNA target site recognition and high-order synaptic assembly are not dependent on canonical DNA but on the position of two flipped-out bases that interact in cis and in trans with the integrase. These extrahelical bases, one of which is required for recombination in vivo, originate from folding of the bottom strand of attC owing to its imperfect internal dyad symmetry. The mechanism reported here supports a new paradigm for how sequence-degenerate single-stranded genetic material is recognized and exchanged between bacteria.     

RNA translocation and unwinding mechanism of HCV NS3 helicase and its coordination by ATP

Helicases are a ubiquitous class of enzymes involved in nearly all aspects of DNA and RNA metabolism. Despite recent progress in understanding their mechanism of action, limited resolution has left inaccessible the detailed mechanisms by which these enzymes couple the rearrangement of nucleic acid structures to the binding and hydrolysis of ATP. Observing individual mechanistic cycles of these motor proteins is central to understanding their cellular functions. Here we follow in real time, at a resolution of two base pairs and 20 ms, the RNA translocation and unwinding cycles of a hepatitis C virus helicase (NS3) monomer. NS3 is a representative superfamily-2 helicase essential for viral replication, and therefore a potentially important drug target. We show that the cyclic movement of NS3 is coordinated by ATP in discrete steps of 11 +/- 3 base pairs, and that actual unwinding occurs in rapid smaller substeps of 3.6 +/- 1.3 base pairs, also triggered by ATP binding, indicating that NS3 might move like an inchworm. This ATP-coupling mechanism is likely to be applicable to other non-hexameric helicases involved in many essential cellular functions. The assay developed here should be useful in investigating a broad range of nucleic acid translocation motors.     

Conjugated action of two species-specific invasion proteins for fetoplacental listeriosis

The ability to cross host barriers is an essential virulence determinant of invasive microbial pathogens. Listeria monocytogenes is a model microorganism that crosses human intestinal and placental barriers, and causes severe maternofetal infections by an unknown mechanism. Several studies have helped to characterize the bacterial invasion proteins InlA and InlB. However, their respective species specificity has complicated investigations on their in vivo role. Here we describe two novel and complementary animal models for human listeriosis: the gerbil, a natural host for L. monocytogenes, and a knock-in mouse line ubiquitously expressing humanized E-cadherin. Using these two models, we uncover the essential and interdependent roles of InlA and InlB in fetoplacental listeriosis, and thereby decipher the molecular mechanism underlying the ability of a microbe to target and cross the placental barrier.     

全面画像而非简单指标

<正>本报告强调:当有关科研人员及其机构的数据被压缩为简单的指标和排名时,有些信息将会遗失。本报告阐述了四类常见分析,如若误用将掩盖真实的科研表现;我们提出了四种可视化选项,用于解读每个度量指标下蕴含的更丰富的信息,以支持开展全面的、负责任的科研管理。我们身边依然存在着声称可通过简单分析来评估论文、科研人员和机构表现的现象。尽管资深分析专家提出反对意见,诸多研究人员对此深表忧虑,但是大学管     

Ecology of Celtis reticulata in Idaho

The small deciduous tree Celtis reticulata (neatleaf hackberry) reaches its northern limit in Idaho, where, contrary to most of its western range, it often occurs as an overstory dominantly. Two hundred fifty stands of this tree were sampled throughout Idaho. Celtis is slow-growing, averaging 4 m tall at 50 yr, and long-lived (to 300-400yr). It occurs in a variety of habitats, from riparian to rocky uplands. Trees grow best where topographically sheltered, such as in draws and narrow canyons, and were soils are loamy. Although plants grow more slowly as surface rock cover increases, stands are often associated with rock, with a mean surface cover of 39% rock. Differences in growth rates were unrelated to parent material and aspect. Most stands are reproducing, in spite of habitat degradation caused by overgrazing, alien plant invasion, and increasing fire frequencies. Stands are typically represented by one dominant cohort; however, young, even-aged stands are rare and are generally found along waterways on stream terraces or at the high-water line. Although slow growing, C. reticulata shows promise for land mangers interested in site enhancement. This native species is long-lived, produces fruit used by wildlife, and provides structural diversity in a semiarid landscape (with a maximum height o 12 m) in areas that are becoming increasingly dominated by exotic plant species.     

Environmental temperature and the growth ofTaenia crassiceps cysticerci in mice

Summary Mice kept at low (5±1 °C) and high (35±1 °C) temperature harboured significantly lessTaenia crassiceps cysticerci than controls kept at 21±1 °C. This effect was more pronounced in heat-stressed than in cold-stressed animals and more in males than in females.