F. A. Van Nieuwenhoven

Fatty-acid-binding protein as a plasma marker for the estimation of myocardial infarct size in humans.

BACKGROUND--There are substantial amounts of cytoplasmic heart-type fatty-acid-binding protein (FABP) (15 kDa) in myocardial tissue. The rapid release of FABP into plasma during ischaemia indicates the possibility of using this protein as a biochemical marker for ischaemic myocardial injury. OBJECTIVE--To study the completeness of the release of FABP from damaged tissue in patients with acute myocardial infarction (AMI) and the suitability of serial plasma FABP concentrations for estimation of myocardial infarct size. METHODS--Immunochemically assayed FABP and enzymatically assayed creatine kinase isoenzyme MB (CK-MB) and alpha-hydroxybutyrate dehydrogenase (HBDH) were determined serially in plasma samples from 49 patients with AMI who had been treated with thrombolytic agents within six hours after the onset of AMI. Previously validated circulatory models and a value of 2.6 h-1 for the fractional clearance rate of FABP from plasma were used to calculate cumulative protein release into plasma. RESULTS--Release of FABP was completed earlier (24-36 h) after AMI than that of CK-MB (50-70 h) and that of HBDH (> 70 h). However, infarct size estimated from the cumulative release of the proteins and expressed as gram equivalents of healthy myocardium per litre of plasma yielded a comparable value of 4-6 for both FABP and the two enzymes. CONCLUSION--The data indicate that FABP released from the heart after AMI is quantitatively recovered in plasma and that FABP is a useful biochemical plasma marker for the estimation of myocardial infarct size in humans.

Cellular fatty acid transport in heart and skeletal muscle as facilitated by proteins.

Despite the importance of long-chain fatty acids (FA) as fuels for heart and skeletal muscles, the mechanism of their cellular uptake has not yet been clarified. There is dispute as to whether FA are taken up by the muscle cellsvia passive diffusion and/or carrier-mediated transport. Kinetic studies of FA uptake by cardiac myocytes and the use of membrane protein-modifying agents have suggested the bulk of FA uptake is due to a protein component. Three membrane-associated FA-binding proteins were proposed to play a role in FA uptake, a 40-kDa plasma membrane FA-binding protein (FABPpm), an 88-kDa FA translocase (FAT/CD36), and a 60-kDa FA transport protein (FATP). In cardiac and skeletal myocytes the intracellular carrier for FA is cytoplasmic heart-type FA-binding protein (H-FABP), which likely transports FA from the sarcolemma to their intracellular sites of metabolism. A scenario is discussed in which FABPpm, FAT/CD36, and H-FABP, probably assisted by an albumin-binding protein, cooperate in the translocation of FA across the sarcolemma.

Connective tissue growth factor and cardiac fibrosis

Cardiac fibrosis is a major pathogenic factor in a variety of cardiovascular diseases and refers to an excessive deposition of extracellular matrix components in the heart, which leads to cardiac dysfunction and eventually overt heart failure. Evidence is accumulating for a crucial role of connective tissue growth factor (CTGF) in fibrotic processes in several tissues including the heart. CTGF orchestrates the actions of important local factors evoking cardiac fibrosis. The central role of CTGF as a matricellular protein modulating the fibrotic process in cardiac remodelling makes it a possible biomarker for cardiac fibrosis and a potential candidate for therapeutic intervention to mitigate fibrosis in the heart.

Role of membrane-associated and cytoplasmic fatty acid-binding proteins in cellular fatty acid metabolism

A number of membrane-associated and cytoplasmic fatty acid-binding proteins (FABPs) are now being implicated in the cellular uptake and intracellular transport of long-chain fatty acids (FA). These proteins each have the capacity of non-covalent binding of FA, are present in tissues actively involved in FA metabolism, and are upregulated in conditions of increased cellular FA metabolism. To date, five distinct membrane FABPs have been described, ranging in mass from 22 to 88 kDa and each showing a characteristic tissue distribution. Evidence for involvement in cellular fatty acid uptake has been provided for several of them, because it was recently found that isolated cell lines transfected with 88-kDa putative fatty acid translocase (FAT; homologous to CD36) or with 63-kDa fatty acid-transport protein show an increased rate of FA uptake. The (at least nine) FABPs of cytoplasmic origin belong to a family of small (14-15 kDa) lipid binding proteins, all having a similar tertiairy structure but differing in binding properties and in tissue occurrence. The biological functions of the various FABPs, possibly exerted in a concerted action among them, comprise solubilization and compartmentalization of FA, facilitation of the cellular uptake and intracellular trafficking of FA, and modulation of mitosis, cell growth, and cell differentiation. In addition, the FABPs have been suggested to participate in and/or modulate FA-mediated signal transduction pathways and FA regulation of gene expression, and to prevent local high FA concentrations thereby contributing to the protection of cells against the toxic effects of FA. In conclusion, long-chain fatty acids are subject to continuous interaction with multiple proteins, which interplay influences their cellular metabolism.

Molecular mechanism of cellular uptake and intracellular translocation of fatty acids

The molecular mechanism of the transport of long-chain fatty acids across cellular membranes and the necessity and precise functioning of specific proteins in this process are still unclear. Various alternative mechanisms have been proposed. Studies with artificial phospholipid bilayers support the concept that fatty acids may enter and traverse the plasma membrane without the involvement of proteins. On the other hand, a number of membrane-associated fatty acid-binding proteins (FABPs) have been described which putatively function as acceptors for fatty acids released from albumin or from lipoproteins. Albumin binding proteins located at the outer cell surface could play an additional role in the delivery of fatty acids. The subsequent transmembrane translocation of fatty acids could take place by a membrane protein acting as a translocase, or by simple diffusion of fatty acids through either the phospholipid bilayer or a pore or channel formed by one or more membrane fatty acid transporters. At the inner side of the plasma membrane, the fatty acid is bound to a cytoplasmic FABP, which serves to buffer the intracellular aqueous fatty acid concentration. The direction of fatty acid migration through the plasma membrane most likely is governed by the transmembrane gradient of fatty acid concentration, assisted to some extent and in selected tissues by co-transport of sodium ions. The intracellular transport of fatty acids from the plasma membrane to the sites of metabolic conversion (oxidation, esterification) or subcellular target (signal transduction) is greatly facilitated by cytoplasmic FABPs. In conclusion, cellular uptake and intracellular translocation of long-chain fatty acids is a multi-step process that is facilitated by various membrane-associated and soluble proteins. The mechanism of cellular uptake of fatty acids probably involves both a passive and carrier-mediated transmembrane translocation.

Membrane-Associated and Cytoplasmic Fatty Acid-Binding Proteins

A number of cellular fatty acid-binding proteins are being implicated in the uptake and intracellular transport of long-chain fatty acids by parenchymal cells. Having been a topic of research for more than 20 years, cytoplasmic fatty acid-binding proteins now are assigned various pivotal functions in intracellular fatty acid transport and metabolism. More recently several membrane-associated fatty acid-binding proteins have been identified and these proteins are thought to function in the transmembrane transport of fatty acids. In this review, a short summary is provided of the latest developments in this research area.

Impaired cardiac functional reserve in type 2 diabetic db/db mice is associated with metabolic, but not structural, remodelling.

Aim:  To identify the initial alterations in myocardial tissue associated with the early signs of diabetic cardiac haemodynamic dysfunction, we monitored changes in cardiac function, structural remodelling and gene expression in hearts of type 2 diabetic db/db mice.

Schwann cell migration and neurite outgrowth are influenced by media conditioned by epineurial fibroblasts.

The regenerative capacity of the peripheral nervous system is largely related to Schwann cells undergoing proliferation and migration after injury and forming growth-supporting substrates for severed axons. Novel data show that fibroblasts to a certain extent regulate the pro-regenerative behavior of Schwann cells. In the setting of peripheral nerve injury, the fibroblasts that form the epineurium come into close contact with both Schwann cells and peripheral axons, but the potential influence on these latter two cell types has not been studied yet. In the present study we explored whether culture media, conditioned by epineurial fibroblasts can influence Schwann cells and/or neurite outgrowth from dorsal root ganglia neurons in vitro. Our data indicate that epineurial fibroblast-conditioned culture media substantially increase Schwann cell migration and the outgrowth of neurites. Schwann cell proliferation remained largely unaffected. These same read-out parameters were assayed in a condition where epineurial fibroblasts were subjected to stretch-cell-stress, a mechanical stressor that plays an important role in traumatic peripheral nerve injuries. Stretch-cell-stress of epineurial fibroblasts did not further change the positive effects of conditioned media on Schwann cell migration and neurite outgrowth. From these data we conclude that an as yet unknown pro-regenerative role can be attributed to epineurial fibroblasts, implying that such cells may affect the outcome of severe peripheral nerve injury.