פרסומים

Bitter taste is one of the basic taste modalities, warning against consuming potential poisons. Bitter compounds activate members of the bitter taste receptor (Tas2r) subfamily of G protein-coupled receptors (GPCRs). The number of functional Tas2rs is species-dependent. Chickens represent an intriguing minimalistic model, because they detect the bitter taste of structurally different molecules with merely three bitter taste receptor subtypes. We investigated the binding modes of several known agonists of a representative chicken bitter taste receptor, ggTas2r1. Because of low sequence similarity between ggTas2r1 and crystallized GPCRs (~10% identity, ~30% similarity at most), the combination of computational approaches with site-directed mutagenesis was used to characterize the agonist-bound conformation of ggTas2r1 binding site between TMs 3, 5, 6 and 7. We found that the ligand interactions with N93 in TM3 and/or N247 in TM5, combined with hydrophobic contacts, are typically involved in agonist recognition. Next, the ggTas2r1 structural model was successfully used to identify three quinine analogues (epiquinidine, ethylhydrocupreine, quinidine) as new ggTas2r1 agonists. The integrated approach validated here may be applicable to additional cases where the sequence identity of the GPCR of interest and the existing experimental structures is low.

Mammary epithelial cells (MEC) secrete fat in the form of milk fat globules (MFG) which are found in milk in diverse sizes. MFG originate from intracellular lipid droplets, and the mechanism underlying their size regulation is still elusive. Two main mechanisms have been suggested to control lipid droplet size. The first is a well-documented pathway, which involves regulation of cellular triglyceride content. The second is the fusion pathway, which is less-documented, especially in mammalian cells, and its importance in the regulation of droplet size is still unclear. Using biochemical and molecular inhibitors, we provide evidence that in MEC, lipid droplet size is determined by fusion, independent of cellular triglyceride content. The extent of fusion is determined by the cell membrane's phospholipid composition. In particular, increasing phosphatidylethanolamine (PE) content enhances fusion between lipid droplets and hence increases lipid droplet size. We further identified the underlying biochemical mechanism that controls this content as the mitochondrial enzyme phosphatidylserine decarboxylase; siRNA knockdown of this enzyme reduced the number of large lipid droplets threefold. Further, inhibition of phosphatidylserine transfer to the mitochondria, where its conversion to PE occurs, diminished the large lipid droplet phenotype in these cells. These results reveal, for the first time to our knowledge in mammalian cells and specifically in mammary epithelium, the missing biochemical link between the metabolism of cellular complex lipids and lipid-droplet fusion, which ultimately defines lipid droplet size.

Even though some progress in diagnosis and treatment has been made over the years, there is still no definitive treatment available for Glioblastoma multiforme (GBM). Convection-enhanced delivery (CED), a continuous infusion-mediated pressure gradient via intracranial catheters, studied in clinical trials, enables in situ drug concentrations several orders of magnitude greater than those achieved by systemic administration. We hypothesized that the currently limited efficacy of CED could be enhanced by a liposomal formulation, thus achieving enhanced drug localization to the tumor site with minimal toxicity. We hereby describe a novel approach for treating GBM by CED of liposomes containing the known chemotherapeutic agent, temozolomide (TMZ). A new technique for encapsulating TMZ in hydrophilic (PEGylated) liposomes, characterized by nano-size (121nm), low polydispersity index (<0.13) and with near-neutral charge (-ʒ,0.2mV), has been developed. Co-infusion of PEGylated Gd-DTPA liposomes and TMZ-liposomes by CED in GBM bearing rats, resulted in enhanced tumor detection with longer residence time than free Gd-DTPA. Treatment of GBM-bearing rats with either TMZ solution or TMZ-liposomes resulted in greater tumor inhibition and significantly higher survival. However, the longer survival and smaller tumor volumes exhibited by TMZ liposomal treatment in comparison to TMZ in solution were insignificant (p<0.053); and only significantly lower edema volumes were observed. Thus, there are no clear-cut advantages to use a liposomal delivery system of TMZ via CED over a drug solution.

Membrane fusion is a central biophysical and biochemical reaction in numerous biological processes. Exocytosis involves the fusion of the secretory vesicle membrane with the plasma membrane in diverse biological systems, including neurotransmitter release at the neuromuscular junction, histamine release from mast cells, chromaffin granule extrusion from adrenal medullary cells, trichocyst discharge in Paramecium, endotoxin-induced degranulation in Limulus amebocytes, and the cortical reaction in sea urchin eggs.1-4 The first stage in the formation of an endocytotic vesicle is the fusion of apposed regions of the invaginated plasma membrane. Later in the endocytotic pathway, endosomes and phagosomes fuse with lysosomes. Receptor recycling back to the plasma membrane proceeds through pinching off of receptor-containing vesicles from the compartment of uncoupling of receptor and ligand. Transport of newly synthesized membrane or secretory proteins from the endoplasmic reticulum to the Golgi apparatus, within the cis, medial and trans regions of the Golgi is thought to be mediated by transport vesicles that bud off from one compartment and fuse with another.5-7. © 1995 by Taylor & Francis.

The Vocabulary The components of a language are எழுத்து ‘sound / letter, சொல் ‘word’ and பொருள் ‘meaning, content’. They are components of the ordinary language and the poetic language; ‘content’ is only of the latter. The poetic language has a symbolic language (செய்யுள்) that is overlaid on the ordinary language (வழக்கு). The above three are not categories of a language, but are its components.

Lipid droplets (LDs) are regulated neutral lipid storage organelles having a central role in numerous cellular processes as well as in various pathologies such as metabolic disorders, immune responses and during pathogen infection. Due to the growing significance of LDs, extensive efforts are made to study the mechanism and the dynamics of their formation and life history and how are these diverted or modified by pathogens. Real-time visualization of lipid droplet biogenesis can assist in clarifying these and other important issues and may have implications towards understanding the pathogenesis of the associated diseases. Typically, LDs are post-experimentally stained using lipophilic dyes and are visualized under a microscope. Alternatively, overexpression of LD-associated proteins or immunofluorescence analyses are used to identify and follow LDs. These experimental approaches only examine a single end point of the experiment and cannot answer questions regarding LD dynamics. Here, we describe a simple and novel experimental setting that allows real-time fluorescence staining and detection of LDs in cultured living as well as infected cells. This method is quick and simple and is not restricted to a specific dye or cell line. Using this system, the biogenesis of LDs and their growth is demonstrated in cells infected with hepatitis C virus (HCV), confirming the strength of this method and the wide range of its applications. © 2017 Elsevier Inc.

Force controlled optical imaging of membranes of living cells is demonstrated. Such imaging has been extended to image membrane potential changes to demonstrate that live cell imaging has been achieved. To accomplish this advance, limitations inherent in atomic force microscopy (AFM) since its inception in 1986 [G. Binnig, C. F. Quate, and C. Gerber, "Atomic Force Microscope," Phys. Rev. Lett. 56, 930-933 (1986).] had to be overcome. The advances allow for live cell imaging of a whole genre of functional biological imaging with stiff (1-10N/m) scanned probe imaging cantilevers. Even topographic imaging of fine cell protrusions, such as microvilli, has been accomplished with such cantilevers. Similar topographic imaging has only recently been demonstrated with the standard soft (0.05N/m) cantilevers that are generally required for live cell imaging. The progress reported here demonstrates both ultrasensitive AFM (~100pN), capable of topographic imaging of even microvilli protruding from cell membranes and new functional applications that should have a significant impact on optical and other approaches in biological imaging of living systems and ultrasoft materials.

 In 1991, the Israeli government introduced emergency legislation canceling the general exit permit that allowed Palestinians to enter Israel. The directive, effective for one year, has been reissued annually ever since, turning the Occupied Territories into a closed military zone. Today, Israel’s permit regime for Palestinians is one of the world’s most extreme and complex apparatuses for population management. Yael Berda worked as a human rights lawyer in Jerusalem and represented more than two hundred Palestinian clients trying to obtain labor permits to enter Israel from the West Bank. With Living Emergency, she brings readers inside the permit regime, offering a first-hand account of how the Israeli secret service, government, and military civil administration control the Palestinian population. Through interviews with Palestinian laborers and their families, conversations with Israeli clerks and officials, and research into the archives and correspondence of governmental organizations, Berda reconstructs the institutional framework of the labyrinthine permit regime, illuminating both its overarching principles and its administrative practices. In an age where terrorism, crime, and immigration are perceived as intertwined security threats, she reveals how the Israeli example informs global homeland security and border control practices, creating a living emergency for targeted populations worldwide. 

 In 1991, the Israeli government introduced emergency legislation canceling the general exit permit that allowed Palestinians to enter Israel. The directive, effective for one year, has been reissued annually ever since, turning the Occupied Territories into a closed military zone. Today, Israel’s permit regime for Palestinians is one of the world’s most extreme and complex apparatuses for population management. Yael Berda worked as a human rights lawyer in Jerusalem and represented more than two hundred Palestinian clients trying to obtain labor permits to enter Israel from the West Bank. With Living Emergency, she brings readers inside the permit regime, offering a first-hand account of how the Israeli secret service, government, and military civil administration control the Palestinian population. Through interviews with Palestinian laborers and their families, conversations with Israeli clerks and officials, and research into the archives and correspondence of governmental organizations, Berda reconstructs the institutional framework of the labyrinthine permit regime, illuminating both its overarching principles and its administrative practices. In an age where terrorism, crime, and immigration are perceived as intertwined security threats, she reveals how the Israeli example informs global homeland security and border control practices, creating a living emergency for targeted populations worldwide. 

Molybdenum trioxide is an interesting inorganic system in which the empty 4d states have potential to hold extra electrons and therefore can change states from insulating opaque (MoO3) to colored semimetallic (HxMoO3). Here, we characterize the local electrogeneration and charge transfer of the synthetic layered two-dimensional 2D MoO3-II (a polymorph of MoO3 and analogous to alpha-MoO3) in response,to two different redox couples; i.e., [Ru(NH3)(6)](3+) and [Fe(CN)(6)](3-) by scanning electrochemical microscopy (SECM). We identify the reduction of [Ru(NH3)(6)](3+) to [Ru(NH3)(6)](2+) at the microelectrode that leads to the reduction of MoO3-11 to conducting blue-colored molybdenum bronze H MoO3. It is recognized that the dominant conduction of the charges occurred preferentially at the edges active sites of the sheets, as edges of the sheets are found to be more conducting. This yields positive feedback current when approaching the microelectrode toward 2D MoO3-II-coated electrode. In contrast, the [Fe(CN)(6)](4-), which is reduced from [Fe(CN)(6)](3-), is found unfavorable to reduce MoO3-II due to its higher redox potential, thus showing a negative feedback current. The charge transfer on MoO3-II is further studied as a function of applied potential. The results shed light on the charge transfer behavior on the surface of MoO3-II coatings and opens the possibility of locally tuning of their oxidation states.