Glycolipid transfer protein and intracellular traffic of glucosylceramide

Summary Glycolipid transfer protein (GL-TP), a nonglycosylated protein with a molecular weight of 22,000 K, has been purified from pig brain. The protein transfers, by a carrier mechanism, glycolipids with a -glucosyl or -galactosyl residue directly linked to either ceramide or diacylglycerol. GL-TP appears to be present in most animal cells, and evidence has been obtained which indicates that it is a cytoplasmic protein. Little is known about the function of GL-TP. Current evidence indicates that glycosphingolipid glycosylation occurs at the luminal side of the Golgi apparatus, except for the glucosylation of ceramide, which has been shown to occur at the cytoplasmic side of the Golgi or endoplasmic membrane. It appears most likely that GL-TP participates in the intracellular traffic of glucosylceramide.     

Cholesteryl ester transfer protein and its inhibition

Cholesteryl ester transfer protein (CETP) is a plasma glycoprotein that facilitates the transfer of cholesteryl esters from the atheroprotective high density lipoprotein (HDL) to the proatherogenic low density lipoprotein cholesterol (LDL) and very low density lipoprotein cholesterol (VLDL) leading to lower levels of HDL but raising the levels of proatherogenic LDL and VLDL. Inhibition of CETP is considered a potential approach to treat dyslipidemia. However, discussions regarding the role of CETP-mediated lipid transfer in the development of atherosclerosis and CETP inhibition as a potential strategy for prevention of atherosclerosis have been controversial. Although many animal studies support the hypothesis that inhibition of CETP activity may be beneficial, negative phase III studies on clinical endpoints with the CETP inhibitor torcetrapib challenged the future perspectives of CETP inhibitors as potential therapeutic agents. The review provides an update on current understanding of the molecular mechanisms involved in CETP activity and its inhibition.     

Autonomous translocation and intracellular trafficking of the cell-penetrating and immune-suppressive effector protein YopM

Extracellular Gram-negative pathogenic bacteria target essential cytoplasmic processes of eukaryotic cells by using effector protein delivery systems such as the type III secretion system (T3SS). These secretion systems directly inject effector proteins into the host cell cytoplasm. Among the T3SS-dependent Yop proteins of pathogenic Yersinia, the function of the effector protein YopM remains enigmatic. In a recent study, we demonstrated that recombinant YopM from Yersinia enterocolitica enters host cells autonomously without the presence of bacteria and thus identified YopM as a novel bacterial cell-penetrating protein. Following entry YopM down-regulates expression of pro-inflammatory cytokines such as tumor necrosis factor α. These properties earmark YopM for further development as a novel anti-inflammatory therapeutic. To elucidate the uptake and intracellular targeting mechanisms of this bacterial cell-penetrating protein, we analyzed possible routes of internalization employing ultra-cryo electron microscopy. Our results reveal that under physiological conditions, YopM enters cells predominantly by exploiting endocytic pathways. Interestingly, YopM was detected free in the cytosol and inside the nucleus. We could not observe any colocalization of YopM with secretory membranes, which excludes retrograde transport as the mechanism for cytosolic release. However, our findings indicate that direct membrane penetration and/or an endosomal escape of YopM contribute to the cytosolic and nuclear localization of the protein. Surprisingly, even when endocytosis is blocked, YopM was found to be associated with endosomes. This suggests an intracellular endosome-associated transport of YopM.     

An attempt to transfer tumor protein specificity to a foreign protein

Résumé L'essai de transférer la spécificité des protéines d'une tumeur à une protéine étrangère par un procédé sérologique, a donné un résultat négatif.     

Interaction of lipid transfer protein with plasma lipoproteins and cell membranes

Summary The hydrophobic lipid components of lipoproteins, cholesteryl ester and triglyceride, are transferred between all lipoproteins by a specific plasma glycoprotein, termed lipid transfer protein (LTP). LTP facilitates lipid transfer by an exchange process in which cholesteryl ester and triglyceride compete for transfer. Thus, LTP promotes remodeling of the lipoprotein structure, and plays an important role in the intravascular metabolism of these particles and in the lipoprotein-dependent pathways of cholesterol clearance from cells. The properties of LTP, its mechanisms of action, its roles in lipoprotein metabolism, and its modes of regulation are reviewed along with recent data that suggest a possible role for this protein in directly modifying cellular lipid composition.     

Role of phospholipid transfer protein in rabbit lung development

A 36-kDa phospholipid transfer protein (PLT-P_R), which preferentially transfers phosphatidyl choline (PC) compared to phosphatidyl inositol (PI), was purified 827-fold from rabbit lung homogenate. Incorporation of cholesterol in unilamellar vesicles reduced the PC transfer activity of PLTP_R. Dipalmitoyl phosphatidyl choline uptake by alveolar type II cells was increased in the presence of the protein, and further enhanced in the presence of surfactant liposomes. However, a decrease in uptake was noted with cholesterol in host membranes. Incorporation of PI into host membranes had a low stimulatory effect on the process. All these effects were more pronounced in adult type II cells compared to premature, term and 3-day-old pups. Received 12 September 2001; accepted 11 October 2001     

Placental transfer of a human IgG2 monoclonal protein

Résumé Étude du passage transplacentaire à son enfant des sousclasses de l'IgG chez une femme atteinte d'une forme benigne de gammapathie monoclonale. Le transfer de la paraprotéine IgG2 L a été établi avec certitude. Il est très probable que les autres sous-classes sont également transferées. Comparée aux résultats publiés sur catabolisme de l'IgG chez l'adulte, la durée de vie de l'IgG total et de l'IgG2 est très longue chez l'enfant.     

Operational filtering of traffic data

An operational filter of traffic state variables is presented for use in designing computer-aided traffic surveillance and control systems. A total of 166 data sets from three traffic surveillance systems were used in the filter development. All the data sets were best represented by an ARIMA (0,1,3) filter. This filter has the following advantages: (1) it yields minimum mean-square-error forecasts if stationarity of the observations can be obtained; (2) it provides much better results than the existing ad hoc filters; (3) it is computationally tractable; and (4) it requires modest computer storage of data. Suggestions and implications for the use of this filter are given.     

Requirement for protein synthesis for the transfer of meiotic induction in a multicellular complex ofBlepharisma

Summary In the ciliateBlepharisma japonicum it is possible to induce meiosis in multicellular homotypic chains. In this work we demonstrate that protein synthesis is required to transfer meiotic activation from one cell to another in a chain.This work was supported by CNR, Programma finalizzato Biologia della Riproduzione.     

Caveolin regulation of neuronal intracellular signaling

Caveolin proteins physically interact with and compartmentalize membrane-localized signaling proteins to facilitate high-fidelity intracellular signaling. Though primarily studied outside the nervous system, recent investigations have revealed that caveolin proteins are key modulators of a variety of neuronal intracellular signaling pathways. Through both protein aggregation and segregation, caveolin proteins can exert positive and negative influences on intracellular signaling. This review will detail recent findings regarding caveolin function in the brain.     

Methods of metal incorporation into intracellular granules

Summary The hepatopancreas of the garden snail (Helix aspersa) contains basophil cells which produce intracellular granules of CaMgP₂O₇. A variety of metals are incorporated into these granules either by direct substitution or by the synthesis of new pyrophosphate material.Supported by NERC grant GR3/3063.     

Control of glass microelectrodes for intracellular recordings

Résumé Une méthode pour vider et examiner des microélectrodes de verre au microscope électronique est décrite. On a trouvé une corrélation entre le diamètre du bout et la résistance électrique. Le diamètre de l'extrémité des électrodes dépend du procédé de remplissement et les meilleurs résultats sont obtenus par la méthode deTasaki modifiée.     

Mechanisms of cellular invasion by intracellular parasites

Numerous disease-causing parasites must invade host cells in order to prosper. Collectively, such pathogens are responsible for a staggering amount of human sickness and death throughout the world. Leishmaniasis, Chagas disease, toxoplasmosis, and malaria are neglected diseases and therefore are linked to socio-economical and geographical factors, affecting well-over half the world’s population. Such obligate intracellular parasites have co-evolved with humans to establish a complexity of specific molecular parasite–host cell interactions, forming the basis of the parasite’s cellular tropism. They make use of such interactions to invade host cells as a means to migrate through various tissues, to evade the host immune system, and to undergo intracellular replication. These cellular migration and invasion events are absolutely essential for the completion of the lifecycles of these parasites and lead to their for disease pathogenesis. This review is an overview of the molecular mechanisms of protozoan parasite invasion of host cells and discussion of therapeutic strategies, which could be developed by targeting these invasion pathways. Specifically, we focus on four species of protozoan parasites Leishmania, Trypanosoma cruzi, Plasmodium, and Toxoplasma, which are responsible for significant morbidity and mortality.     

Effect of body temperature on intracellular pH

Zusammenfassung Zwischen dem extrazellulären pH und dem pH des gesamten Intrazellulärraumes besteht eine annähernd lineare Beziehung. Senkung der Körpertemperatur führt zu einer Erhöhung des pH _i , hat aber keinen Einfluss auf den Regressionskoeffizienten und damit auf die intrazelluläre Pufferkapazität.     

Cell density-induced changes in lipid composition and intracellular trafficking

Cell density is one of the extrinsic factors to which cells adapt their physiology when grown in culture. However, little is known about the molecular changes which occur during cell growth and how cellular responses are then modulated. In many cases, inhibitors, drugs or growth factors used for in vitro studies change the rate of cell proliferation, resulting in different cell densities in control and treated samples. Therefore, for a comprehensive data analysis, it is essential to understand the implications of cell density on the molecular level. In this study, we have investigated how lipid composition changes during cell growth, and the consequences it has for transport of Shiga toxin. By quantifying 308 individual lipid species from 17 different lipid classes, we have found that the levels and species distribution of several lipids change during cell growth, with the major changes observed for diacylglycerols, phosphatidic acids, cholesterol esters, and lysophosphatidylethanolamines. In addition, there is a reduced binding and retrograde transport of Shiga toxin in high density cells which lead to reduced intoxication by the toxin. In conclusion, our data provide novel information on how lipid composition changes during cell growth in culture, and how these changes can modulate intracellular trafficking.     

Knowledge transfer and its contexts

Knowledge transfer across different contexts has become an increasingly prevalent feature of current science. As such, it is a relevant topic also for historians and philosophers of science. This special issue presents a set of papers that study knowledge transfer in various disciplines. The contributions approach the topic from 1) an integrated history and philosophy of science perspective, 2) a systematic philosophical perspective, or 3) a historical perspective. Taken together, they give a broad introduction into the topic and offer a set of conceptual resources for the study of knowledge transfer in multiple contexts.     

31P-NMR study on nucleotides and intracellular pH of hereditary spherocytes

Summary As determined by³¹p-NMR spectroscopy, intracellarar pH of hereditary spherocytes was lower (pH 6.7–6.9) than that of normal red cells. The level of adenosine diphosphate in hereditary spherocytes was found to be persistently high. The metabolism of nucleotides and other phosphoryl compounds in human red blood cells have been studied in detail by³¹p-NMR spectroscopy^1–3. However, to our knowledge, there seems to be no report describing the result of³¹p-NMR spectroscopy on red blood cells from hereditary spherocytosis.