scholarly journals SCM, a novel M-like protein from Streptococcus canis, binds (mini)-plasminogen with high affinity and facilitates bacterial transmigration

2011 ◽  
Vol 434 (3) ◽  
pp. 523-535 ◽  
Author(s):  
Marcus Fulde ◽  
Manfred Rohde ◽  
Angela Hitzmann ◽  
Klaus T. Preissner ◽  
D. Patric Nitsche-Schmitz ◽  
...  
Keyword(s):  
Catalytic Domain ◽  
Time Lapse ◽  
Dot Blot ◽  
Electron Microscopic ◽  
Bacterial Surface ◽  
Amino Acid Analogues ◽  
Human Plasminogen ◽  
Streptococcus Canis ◽  
Time Lapse Imaging ◽  
First Time

Streptococcus canis is an important zoonotic pathogen capable of causing serious invasive diseases in domestic animals and humans. In the present paper we report the binding of human plasminogen to S. canis and the recruitment of proteolytically active plasmin on its surface. The binding receptor for plasminogen was identified as a novel M-like protein designated SCM (S. canis M-like protein). SPR (surface plasmon resonance) analyses, radioactive dot-blot analyses and heterologous expression on the surface of Streptococcus gordonii confirmed the plasminogen-binding capability of SCM. The binding domain was located within the N-terminus of SCM, which specifically bound to the C-terminal part of plasminogen (mini-plasminogen) comprising kringle domain 5 and the catalytic domain. In the presence of urokinase, SCM mediated plasminogen activation on the bacterial surface that was inhibited by serine protease inhibitors and lysine amino acid analogues. Surface-bound plasmin effectively degraded purified fibrinogen as well as fibrin clots, resulting in the dissolution of fibrin thrombi. Electron microscopic illustration and time-lapse imaging demonstrated bacterial transmigration through fibrinous thrombi. The present study has led, for the first time, to the identification of SCM as a novel receptor for (mini)-plasminogen mediating the fibrinolytic activity of S. canis.

scholarly journals Fluctuation in radioresponse of HeLa cells during the cell cycle evaluated based on micronucleus frequency

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Hiroaki Shimono ◽  
Atsushi Kaida ◽  
Hisao Homma ◽  
Hitomi Nojima ◽  
Yusuke Onozato ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Hela Cells ◽  
Time Lapse ◽  
S Phase ◽  
G1 Phase ◽  
M Phase ◽  
G2 Phase ◽  
The Difference ◽  
Time Lapse Imaging ◽  
First Time

AbstractIn this study, we examined the fluctuation in radioresponse of HeLa cells during the cell cycle. For this purpose, we used HeLa cells expressing two types of fluorescent ubiquitination-based cell cycle indicators (Fucci), HeLa-Fucci (CA)2 and HeLa-Fucci (SA), and combined this approach with the micronucleus (MN) assay to assess radioresponse. The Fucci system distinguishes cell cycle phases based on the colour of fluorescence and cell morphology under live conditions. Time-lapse imaging allowed us to further identify sub-positions within the G1 and S phases at the time of irradiation by two independent means, and to quantitate the number of MNs by following each cell through M phase until the next G1 phase. Notably, we found that radioresponse was low in late G1 phase, but rapidly increased in early S phase. It then decreased until late S phase and increased in G2 phase. For the first time, we demonstrated the unique fluctuation of radioresponse by the MN assay during the cell cycle in HeLa cells. We discuss the difference between previous clonogenic experiments using M phase-synchronised cell populations and ours, as well as the clinical implications of the present findings.

The preparatory phase of the Mw 6.1 2009 L’Aquila (Italy) normal faulting earthquake traced by foreshock time-lapse tomography

Geology ◽  
2019 ◽  
Vol 48 (1) ◽  
pp. 49-55 ◽  
Author(s):  
Paola Baccheschi ◽  
Pasquale De Gori ◽  
Fabio Villani ◽  
Fabio Trippetta ◽  
Claudio Chiarabba
Keyword(s):  
Central Italy ◽  
Fluid Pressure ◽  
Time Lapse ◽  
Normal Faulting ◽  
S Wave ◽  
Preparatory Phase ◽  
Major Fault ◽  
Time Lapse Imaging ◽  
First Time ◽  
Locked Fault

Abstract The Mw 6.1 (6 April 2009) L’Aquila (Italy) earthquake occurred in one of the most seismically active areas of central Italy and was preceded by a three-month-long foreshock period. Thanks to recordings by a regional permanent network, we derive for the first time P- and S-wave velocity tomographic models of a major fault prone to an imminent main shock. Close to the Mw 6.1 hypocenter, we observe high Vp (>6.8 km/s) and high Vp/Vs (>1.9) consistent with thick dolomitic volumes filled with fluids sealed by impermeable anhydritic layers. Significant changes in velocities defined by time-lapse imaging during the foreshock period suggest rapid fluid migration through the locked fault zone. The complex positive feedback between fluid pressure buildup and hydrofracturing of the dolomitic reservoir, testified by foreshock production, eventually provoked the catastrophic coseismic breaching of the fault seal. Our results show that foreshock time-lapse tomography provides clues on the preparatory phase of a large normal-faulting earthquake.

scholarly journals Cooperative Plasminogen Recruitment to the Surface of Streptococcus canis via M Protein and Enolase Enhances Bacterial Survival

mBio ◽  
2013 ◽  
Vol 4 (2) ◽  
Author(s):  
Marcus Fulde ◽  
Manfred Rohde ◽  
Andy Polok ◽  
Klaus T. Preissner ◽  
Gursharan Singh Chhatwal ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Human Neutrophils ◽  
Synergistic Activity ◽  
Bacterial Survival ◽  
Content Type ◽  
Bacterial Surface ◽  
M Proteins ◽  
Human Plasminogen ◽  
Streptococcus Canis ◽  
Phagocytic Killing

ABSTRACTStreptococcus canisis a zoonotic pathogen capable of causing serious invasive diseases in domestic animals and humans. Surface-exposed M proteins and metabolic enzymes have been characterized as major virulence determinants in various streptococcal species. Recently, we have identified SCM, the M-like protein ofS. canis, as the major receptor for miniplasminogen localized on the bacterial surface. The present study now characterizes the glycolytic enzyme enolase as an additional surface-exposed plasminogen-binding protein. According to its zoonotic properties, purifiedS. canisenolase binds to both human and canine plasminogen and facilitates degradation of aggregated fibrin matrices after activation with host-derived urokinase-type plasminogen activator (uPA). Unlike SCM, which binds to the C terminus of human plasminogen, theS. canisenolase interacts N terminally with the first four kringle domains of plasminogen, representing angiostatin. Radioactive binding analyses confirmed cooperative plasminogen recruitment to both surface-exposed enolase and SCM. Furthermore, despite the lack of surface protease activity via SpeB inS. canis, SCM is released and reassociated homophilically to surface-anchored SCM and heterophilically to surface-bound plasminogen. In addition to plasminogen-mediated antiphagocytic activity, reassociation of SCM to the bacterial surface significantly enhanced bacterial survival in phagocytosis analyses using human neutrophils.IMPORTANCEStreptococcal infections are a major issue in medical microbiology due to the increasing spread of antibiotic resistances and the limited availability of efficient vaccines. Surface-exposed glycolytic enzymes and M proteins have been characterized as major virulence factors mediating pathogen-host interaction. Since streptococcal infection mechanisms exert a subset of multicombinatorial processes, the investigation of synergistic activities mediated via different virulence factors has become a high priority. Our data clearly demonstrate that plasminogen recruitment to theStreptococcus canissurface via SCM and enolase in combination with SCM reassociation enhances bacterial survival by protecting against phagocytic killing. These data propose a new cooperative mechanism for prevention of phagocytic killing based on the synergistic activity of homophilic and heterophilic SCM binding in the presence of human plasminogen.

Morphokinetics of early equine embryo development in vitro using time-lapse imaging, and use in selecting blastocysts for transfer

2019 ◽  
Vol 31 (12) ◽  
pp. 1851 ◽  
Author(s):  
Niamh Lewis ◽  
Karen Schnauffer ◽  
Katrin Hinrichs ◽  
Monica Morganti ◽  
Stephen Troup ◽  
...  
Keyword(s):  
Embryo Development ◽  
Time Lapse ◽  
Developmental Competence ◽  
Early Cleavage ◽  
Blastocyst Development ◽  
Use Of Time ◽  
Development In Vitro ◽  
Time Lapse Imaging ◽  
First Time

The use of time-lapse imaging (TLI) in the evaluation of morphokinetics associated with invitro developmental competence is well described for human, cattle and pig embryos. It is generally accepted that embryos that complete early cleavage sooner are more likely to form blastocysts and that timing of later events, such as blastocyst formation and expansion, are predictive of implantation potential and euploid status. In the horse, morphokinetics as a predictor of developmental competence has received little attention. In this study we evaluated the morphokinetics of early equine embryo development invitro for 144 oocytes after intracytoplasmic sperm injection and report the timings of blastocyst development associated with ongoing pregnancy for the first time. There was a tendency for time of cytoplasmic extrusion and first cleavage to occur earlier in the embryos that went on to form blastocysts (n=19) compared with those that arrested, and for first cleavage to occur earlier in blastocysts that established pregnancies that were ongoing (n=4) compared with pregnancies that were lost (n=2). TLI was clinically useful in identifying blastocysts when evaluation of morphology on static imaging was equivocal.

Annulate lamellae in the Sertoli cells of guinea pig testis after unilateral torsion of the spermatic cord

1984 ◽  
Vol 42 ◽  
pp. 196-197
Author(s):  
J. Chakraborty ◽  
A. P. Sinha Hikim ◽  
J. S. Jhunjhunwala
Keyword(s):  
Sertoli Cells ◽  
Guinea Pigs ◽  
Spermatic Cord ◽  
Present Report ◽  
Cell Types ◽  
Annulate Lamellae ◽  
Spermatogenic Cells ◽  
Electron Microscopic ◽  
Structural Details ◽  
First Time

Although the presence of annulate lamellae was noted in many cell types, including the rat spermatogenic cells, this structure was never reported in the Sertoli cells of any rodent species. The present report is based on a part of our project on the effect of torsion of the spermatic cord to the contralateral testis. This paper describes for the first time, the fine structural details of the annulate lamellae in the Sertoli cells of damaged testis from guinea pigs.One side of the spermatic cord of each of six Hartly strain adult guinea pigs was surgically twisted (540°) under pentobarbital anesthesia (1). Four months after induction of torsion, animals were sacrificed, testes were excised and processed for the light and electron microscopic investigations. In the damaged testis, the majority of seminiferous tubule contained a layer of Sertoli cells with occasional spermatogonia (Fig. 1). Nuclei of these Sertoli cells were highly pleomorphic and contained small chromatinic clumps adjacent to the inner aspect of the nuclear envelope (Fig. 2).

A microscopic marker experiment study on high temperature oxidation of Ni-Al alloy

1986 ◽  
Vol 44 ◽  
pp. 514-515
Author(s):  
Shou-kong Fan
Keyword(s):  
Transport Mechanism ◽  
High Temperature Oxidation ◽  
Transverse Section ◽  
Al Alloy ◽  
Metal Interface ◽  
Electron Microscopic ◽  
Temperature Oxidation ◽  
Analytical Electron ◽  
Microscopic Studies ◽  
First Time

Transmission and analytical electron microscopic studies of scale microstructures and microscopic marker experiments have been carried out in order to determine the transport mechanism in the oxidation of Ni-Al alloy. According to the classical theory, the oxidation of nickel takes place by transport of Ni cations across the scale forming new oxide at the scale/gas interface. Any markers deposited on the Ni surface are expected to remain at the scale/metal interface after oxidation. This investigation using TEM transverse section techniques and deposited microscopic markers shows a different result,which indicates that a considerable amount of oxygen was transported inward. This is the first time that such fine-scale markers have been coupled with high resolution characterization instruments such as TEM/STEM to provide detailed information about evolution of oxide scale microstructure.

Cleavage of Human Embryos: Options and Diversity

Acta Naturae ◽  
2016 ◽  
Vol 8 (3) ◽  
pp. 88-96
Author(s):  
Yu. K. Doronin ◽  
I. V. Senechkin ◽  
L. V. Hilkevich ◽  
M. A. Kurcer
Keyword(s):  
Time Lapse ◽  
Reproductive Technologies ◽  
Human Embryos ◽  
Imaging Data ◽  
Noninvasive Methods ◽  
Topographic Features ◽  
Time Parameters ◽  
Embryo Cleavage ◽  
Time Lapse Imaging ◽  
Developmental Dynamics

In order to estimate the diversity of embryo cleavage relatives to embryo progress (blastocyst formation), time-lapse imaging data of preimplantation human embryo development were used. This retrospective study is focused on the topographic features and time parameters of the cleavages, with particular emphasis on the lengths of cleavage cycles and the genealogy of blastomeres in 2- to 8-cell human embryos. We have found that all 4-cell human embryos have four developmental variants that are based on the sequence of appearance and orientation of cleavage planes during embryo cleavage from 2 to 4 blastomeres. Each variant of cleavage shows a strong correlation with further developmental dynamics of the embryos (different cleavage cycle characteristics as well as lengths of blastomere cycles). An analysis of the sequence of human blastomere divisions allowed us to postulate that the effects of zygotic determinants are eliminated as a result of cleavage, and that, thereafter, blastomeres acquire the ability of own syntheses, regulation, polarization, formation of functional contacts, and, finally, of specific differentiation. This data on the early development of human embryos obtained using noninvasive methods complements and extend our understanding of the embryogenesis of eutherian mammals and may be applied in the practice of reproductive technologies.

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