Karen Dell: iBiology:Meet the world's best biologists through the Internet
Karen Dell来自美国细胞生物学学会，她将简述通过iBiology来获取生物学学习和交流的资源。
李于：SIRT1 Regulation of Energy Metabolism: Attenuation of hepatic Steatosis and Obesity
Fibroblast growth factor 21 (FGF21) is the hepatocyte-derived hormone that regulates fatty acid metabolism and has potential to treat obesity and diabetes. We recently indicate that hepatic overexpression of SIRT1 in diabetic mice attenuates hepatic steatosis and insulin resistance. However, the in vivo long-term consequence of hepatic SIRT1 ablation in liver physiology remains unknown.
We showed that hepatocyte-specific SIRT1 knockout (SIRT1 LKO) mice with the albumin Cre-loxP system exhibited a striking phenotype with greater propensity for obesity on a chow diet, characterized by increased whole body mass and fat mass, reduced energy expenditure, and unaltered food intake and physical activity. The obese phenotypes of SIRT1 LKO mice were associated with reduced hepatic and circulating levels of fasting FGF21.
hepatic impairment of FGF21 repressed expression of key enzymes involving fatty acid oxidation such as CPT1α and MCAD, and inhibited expression of ketogenic enzymes including ACAT1, HMGCS2, HMGCL, and BDH1, thereby reducing plasma β–hydroxybutyrate levels in SIRT1 LKO mice. Moreover, transcriptional activity of a FGF21 promoter-driven luciferase reporter was stimulated by SIRT1 activators, resveratrol and SRT1720, in SIRT1+/+ MEFs, but not in SIRT1-/- MEFs.
The ability of resveratrol and SRT1720 to stimulate FGF21 protein was abolished by SIRT1 H335A inactive mutant or by nicotinamide and splitomicin in hepG2 cells. Induction of FGF21 by SIRT1 activators enhanced expression of key enzymes for fatty acid oxidation and ketogenesis.
These in vivo and in vitro findings characterize 1) hepatic SIRT1 as a master regulator of FGF21; 2) SIRT1-dependent activation of FGF21 in liver as a component for adaptive fasting response; and 3) defective hepatic SIRT1 and FGF21 signaling as a key pathological determinant of energy metabolic abnormality and obesity susceptibility.
Western Blot Using The invitrogen NuPAGE Novex Bis-Tris MiniGel System(Aubin Penna.Ph.D)
Western Blot Using The invitrogen NuPAGE Novex Bis-Tris MiniGel System(Aubin Penna.Ph.D)
Immunoblot Analysis Sean Gallagher(UVP,LLC)and Deb Chakravart(Proteomic Center)
Immunoblot Analysis Sean Gallagher(UVP,LLC)and Deb Chakravart(Proteomic Center)
病毒结构的一般原则 - Stephen Harrison P1
本视频由科普中国和生物医学大讲堂出品
Stephen Harrison (Harvard) Part 1: Virus structures: General principles
Harrison begins his talk by asking why most non-enveloped viruses and some enveloped viruses are symmetrical in shape. he proceeds to show us lovely images of the structures obtained by x-ray crystallography of numerous viral coat proteins. Deciphering these structures allowed scientists to understand that viral coat proteins form multimers, such as dimers and pentamers, which in turn interact with a scaffold that ensures that the coat proteins are correctly placed. This arrangement results in symmetrically shaped viruses.
In Part 1, Harrison also explains that enveloped viruses infect cells by inducing the fusion of the viral and host cell membranes. he delves deeper into the molecular mechanism of membrane fusion driven by the hemagglutinin or HA protein of the influenza virus in Part 2 of his talk.
Non-enveloped viruses, on the other hand, must enter cells by a mechanism other than membrane fusion. This is the focus of Part 3. Using rotavirus as a model, Harrison and his colleagues have used a combination of Xray crystallography and electron cryomicroscopy to decipher how the spike protein on the viral surface changes its conformation and perforates the cell membrane allowing the virus to enter the cell.
病毒的膜融合 - Stephen Harrison P2
本视频由科普中国和生物医学大讲堂出品
Stephen Harrison (Harvard) Part 2: Viral membrane fusion
Harrison begins his talk by asking why most non-enveloped viruses and some enveloped viruses are symmetrical in shape. he proceeds to show us lovely images of the structures obtained by x-ray crystallography of numerous viral coat proteins. Deciphering these structures allowed scientists to understand that viral coat proteins form multimers, such as dimers and pentamers, which in turn interact with a scaffold that ensures that the coat proteins are correctly placed. This arrangement results in symmetrically shaped viruses.
In Part 1, Harrison also explains that enveloped viruses infect cells by inducing the fusion of the viral and host cell membranes. he delves deeper into the molecular mechanism of membrane fusion driven by the hemagglutinin or HA protein of the influenza virus in Part 2 of his talk.
Non-enveloped viruses, on the other hand, must enter cells by a mechanism other than membrane fusion. This is the focus of Part 3. Using rotavirus as a model, Harrison and his colleagues have used a combination of Xray crystallography and electron cryomicroscopy to decipher how the spike protein on the viral surface changes its conformation and perforates the cell membrane allowing the virus to enter the cell.
非包膜病毒如何侵入细胞 - Stephen Harrison P3
本视频由科普中国和生物医学大讲堂出品
Stephen Harrison (Harvard) Part 3: Non-enveloped virus entry
Harrison begins his talk by asking why most non-enveloped viruses and some enveloped viruses are symmetrical in shape. he proceeds to show us lovely images of the structures obtained by x-ray crystallography of numerous viral coat proteins. Deciphering these structures allowed scientists to understand that viral coat proteins form multimers, such as dimers and pentamers, which in turn interact with a scaffold that ensures that the coat proteins are correctly placed. This arrangement results in symmetrically shaped viruses.
In Part 1, Harrison also explains that enveloped viruses infect cells by inducing the fusion of the viral and host cell membranes. he delves deeper into the molecular mechanism of membrane fusion driven by the hemagglutinin or HA protein of the influenza virus in Part 2 of his talk.
Non-enveloped viruses, on the other hand, must enter cells by a mechanism other than membrane fusion. This is the focus of Part 3. Using rotavirus as a model, Harrison and his colleagues have used a combination of Xray crystallography and electron cryomicroscopy to decipher how the spike protein on the viral surface changes its conformation and perforates the cell membrane allowing the virus to enter the cell.
病毒与宿主细胞表面结合的方式 - Ari helenius P1
本视频由科普中国和生物医学大讲堂出品
Ari helenius (ETH Zurich) Part 1: Virus entry
Viruses are extremely simple and small yet they are responsible for many of the worlds diseases. A virus particle consists of only a genome, a protein coat or capsid, and sometimes a surrounding lipid envelope. To replicate, a virus must successfully enter a host cell, uncoat its genome, and appropriate the host cell machinery to replicate its genome and produce viral proteins. Part 1 of this lecture will discuss ways in which viruses bind to the surface of host cells. Simian Virus 40 which binds to specific cell surface glycolipids, and Human Papilloma Virus-16 which binds to sites on filoipodia, are examples of different binding mechanisms. Attachment of viruses to the plasma membrane activates cell signaling resulting in endocytosis of the viral particles. This lecture is appropriate for upper level undergraduate and graduate classes studying virology or endocytosis.
牛痘病毒如何进入细胞 - Ari helenius P3
本视频由科普中国和生物医学大讲堂出品
Ari helenius (ETH Zurich) Part 3: Open Sesame: Cell Entry and Vaccinia Virus
Part 3 focuses on a single virus, the Vaccinia virus, as a model for cell binding, signaling and endocytosis. Fluorescently labeled Vaccinia viruses bind to and surf along host cell filopodia. helenius lab members noticed that when Vaccinia, unlike other viruses, reached the surface of the cell body it caused the plasma membrane to form blebs. Further experiments showed that the virus tricks the cell into thinking it is apoptotic debris. This induces blebbing and subsequent uptake of the virus by macropinocytosis. Additionally, automated high throughput siRNA screening was used to screen a large number of infected cells for host genes required for Vaccinia virus uptake. Analysis of the genes identified allowed host factors and processes critical to viral infection to be identified. Expansion of this technique may provide a new source of information on pathogen-host interactions.