High-density lipoproteins (HDLs) protect against atherosclerosis by removing excess cholesterol from macrophages through the ATP-binding cassette transporter A1 (ABCA1) and ATP-binding cassette transporter G1 (ABCG1) pathways involved in reverse cholesterol transport. Factors that impair the availability of functional apolipoproteins or the activities of ABCA1 and ABCG1 could, therefore, strongly influence atherogenesis. HDL also inhibits lipid oxidation, restores endothelial function, exerts anti-inflammatory and antiapoptotic actions, and exerts anti-inflammatory actions in animal models. Such properties could contribute considerably to the capacity of HDL to inhibit atherosclerosis. Systemic and vascular inflammation has been proposed to convert HDL to a dysfunctional form that has impaired antiatherogenic effects. A loss of anti-inflammatory and antioxidative proteins, perhaps in combination with a gain of proinflammatory proteins, might be another important component in rendering HDL dysfunctional. The proinflammatory enzyme myeloperoxidase induces both oxidative modification and nitrosylation of specific residues on plasma and arterial apolipoprotein A-I to render HDL dysfunctional, which results in impaired ABCA1 macrophage transport, the activation of inflammatory pathways, and an increased risk of coronary artery disease. Understanding the features of dysfunctional HDL or apolipoprotein A-I in clinical practice might lead to new diagnostic and therapeutic approaches to atherosclerosis.

The scavenger receptor class B, type I (SR-BI), binds high density lipoprotein (HDL) and mediates selective uptake of cholesteryl ester from HDL and HDL-dependent cholesterol efflux from cells. We recently identified a new mRNA variant that differs from the previously characterized form in that the encoded C-terminal cytoplasmic domain is almost completely different. In the present study, we demonstrate that the mRNAs for mouse SR-BI and SR-BII (previously termed SR-BI.2) are the alternatively spliced products of a single gene. The translation products predicted from human, bovine, mouse, hamster, and rat cDNAs exhibit a high degree of sequence similarity within the SR-BII C-terminal domain (62-67% identity when compared with the human sequence), suggesting that this variant is biologically important. SR-BII protein represents approximately 12% of the total immunodetectable SR-BI/II protein in mouse liver. Subcellular fractionation of transfected Chinese hamster ovary cells showed that SR-BII, like SR-BI, is enriched in caveolae, indicating that the altered cytoplasmic tail does not affect targeting of the receptor. SR-BII mediated both selective cellular uptake of cholesteryl ether from HDL as well as HDL-dependent cholesterol efflux from cells, although with approximately 4-fold lower efficiency than SR-BI. In vivo studies using adenoviral vectors showed that SR-BII was relatively less efficient than SR-BI in reducing plasma HDL cholesterol. These studies show that SR-BII, an HDL receptor isoform containing a distinctly different cytoplasmic tail, mediates selective lipid transfer between HDL and cells, but with a lower efficiency than the previously characterized variant.

The liver is the most common site of metastatic disease1. While this metastatic tropism may reflect mechanical trapping of circulating tumor cells, liver metastasis is also dependent, at least in part, on formation of a “pro-metastatic” niche that supports tumor cell spread to the liver2,3. Mechanisms that direct formation of this niche, though, are poorly understood. Here, we show that hepatocytes coordinate myeloid cell accumulation and fibrosis within the liver, and in doing so, increase the susceptibility of the liver to metastatic seeding and outgrowth. Early during pancreatic tumorigenesis, hepatocytes demonstrate activation of Signal Transducer and Activator of Transcription 3 (STAT3) signaling and increased production of serum amyloid A1 and A2 (SAA). Overexpression of SAA by hepatocytes also occurs in pancreatic and colorectal cancer patients with liver metastases, and many patients with locally advanced and metastatic disease display elevated levels of circulating SAA. STAT3 activation in hepatocytes and the subsequent production of SAA are dependent on interleukin 6 (IL-6) that is released into the circulation by non-malignant cells. Genetic ablation or blockade of components of IL-6/STAT3/SAA signaling prevents establishment of a pro-metastatic niche and inhibits liver metastasis. Our data reveal an intercellular network underpinned by hepatocytes that forms the basis for a pro-metastatic niche in the liver and identify new therapeutic targets.

Transforming growth factors (TGFs) are mitogenic polypeptides produced most conspicuously by transformed cells and conferring on normal cells several phenotypic alterations associated with transformation. TGFs comprise two distinct sets of molecules: TGF-alpha s are structurally similar to epidermal growth factor (EGF), binding to and inducing the tyrosine phosphorylation of the EGF receptor in a manner indistinguishable from that of EGF. In addition, the 50-amino acid rat TGF-alpha has 33 and 44% homologies with mouse and human EGFs, respectively, and shares with EGFs a conserved pattern of three disulphide bridges. Thus, it has been proposed that TGF-alpha s belong to a family of EGF-like polypeptides. TGF-beta s, on the other hand, display no measurable binding to EGF receptors, but potentiate the growth-stimulating activities of TGF-alpha. Here we report the isolation of a complementary DNA clone encoding rat TGF-alpha. This cDNA hybridizes to a 4.5-kilobase (kb) messenger RNA that is 30 times larger than necessary to code for a 50-amino acid polypeptide and is present not only in retrovirus-transformed rat cells but also at lower levels in normal rat tissues. The nucleotide sequence of the cDNA predicts that TGF-alpha is synthesized as a larger product and that the larger form may exist as a transmembrane protein. However, unlike many polypeptide hormones (including EGF), cleavage of the 50-amino acid TGF-alpha from the larger form does not occur at paired basic residues, but rather between alanine and valine residues, suggesting the role of a novel protease.

Thomas Campbell and colleagues report findings of a randomized trial conducted in multiple countries regarding the efficacy of antiretroviral regimens with simplified dosing.

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Recombinant type 1 transforming growth factor beta (TGF-,) was expressed to high levels in CHO cells by using dihydrofolate reductase (dhfr) gene amplification. The expression plasmid was derived from the pSV2 vectors and contained, in tandem, the simian TGF-I and mouse dhfr cDNAs. Transcription of both cDNAs was controlled by the simian virus 40 early promoter. Stepwise selection of transfected CHO cells in increasing concentrations of methotrexate yielded cell lines that expressed amplified TGF-, nucleic acid sequences. The expression plasmid DNA was amplified greater than 35-fold in one of the methotrexate-selected transfectants.The major proteins secreted by these cells consisted of latent as judged by immunoblots by using site-specific anti-peptide antibodies derived from various regions of the TGF-, precursor. Levels of recombinant TGF-0 protein secreted by these cells approached 30 ,ug/24 h per 107 cells and required prior acidification for optimal activity; nonacidified supernatants were approximately 1% as active as acidified material. Antibodies directed toward sequences present in the mature growth factor readily identified a proteolytically processed recombinant TGF-, which, on sodium dodecyl sulfate-polyacrylamide gels, comigrated with highly purified natural TGF-4. In addition to mature recombinant TGF-jA, site-specific antibodies demonstrated the existence of larger TGF-3 precursor polypeptides. The availability of biologically active recombinant type 1 TGF-I8 and precursor forms should provide a means to examine the structure, function, and potential in vivo therapeutic use of this growth factor.Results of recent studies have suggested that transforming growth factor beta (TGF-P) consists of a family of closely related biologically active polypeptides with potent cellularmodulating activities. Three molecular forms of TGF-P (types 1, 2 and 1.2) have been identified and characterized.Type 1 TGF-P was the first growth factor isolated and, as a result, the best characterized (for reviews, see references 23 and 47). sources have been obtained. The predicted protein sequences derived from the cDNAs reveal a remarkable homology. The mature forms of the human and simian growth factor differ from the murine factor by one amino acid. Analysis of the various TGF-,B cDNA clones suggests that the mature 112 amino acid chain of type 1 TGF-P is derived from a much larger precursor polypeptide by proteolytic cleavage. The full-length precursor of TGF-,1 shows a high degree of structural conservation and constitutes 391 amino acids for human and 390 amino acids for both murine and simian precursors. A typical leader sequence is observed in the precursor, as well as three potential N-linked glycosylation sites. A dibasic sequence immediately precedes the amino-terminal alanine residue of the mature growth factor, indicating the involvement of a dibasic protease in processing of the growth factor.To understand in more detail the structure and processing of type 1 TGF-P, as well as to provide a source for large...

Objective-The purpose of this study was to examine the interactive action of serum amyloid A (SAA), group IIA secretory phospholipase A 2 (sPLA 2 -IIA), and cholesteryl ester transfer protein (CETP) on HDL remodeling and cholesterol efflux during the acute phase (AP) response elicited in humans after cardiac surgery. Methods and Results-Plasma was collected from patients before (pre-AP), 24 hours after (AP-1 d), and 5 days after cardiac surgery (AP-5 d). SAA levels were increased 16-fold in AP-1 d samples. Key Words: SAA Ⅲ HDL Ⅲ CETP Ⅲ apoA-I Ⅲ inflammation I nflammation induces major changes in HDL levels and composition. Mediators of inflammation such as tumor necrosis factor (TNF)-␣ and interleukin (IL)-6 induce expression of serum amyloid A 1 and group IIA secretory phospholipase A 2 (sPLA 2 -IIA), 2 which dramatically alter HDL apolipoprotein content and levels, respectively. Acute phase SAA in the plasma is associated with HDL, where it can comprise the major apolipoprotein. 3 The increase in sPLA 2 -IIA activity results in hydrolysis of HDL surface phospholipids and a decrease in HDL particle size. 4 The plasma cholesteryl ester transfer protein (CETP) is an integral component of reverse cholesterol transport and regulates HDL cholesterol concentrations. By promoting the transfer of cholesteryl esters (CE) from HDL to apoB-containing lipoprotein particles, HDL-derived CE is taken up via the LDL receptor and cleared by the liver. 5 An additional result of CETP action is the generation of lipid-poor apoA-I, 6 a key acceptor in ATP-binding cassette transporter AI (ABCA1)-mediated lipid efflux. 7 The presence of SAA on HDL holds the potential to impact both the CE transfer and the apoA-I liberating ability of CETP. sPLA 2 -IIA could also impact the latter action of CETP as apoA-I was shown to dissociate more readily from CETP-remodeled reconstituted HDL after hydrolysis by bee venom phospholipase A 2 . 8 Given the centrality of inflammation in atherogenesis, there is a paucity of information regarding CETP function when acute phase HDL is the "substrate." In the present study, we used plasma from patients undergoing cardiac surgery with cardiopulmonary bypass as a "standardized" insult where the oxygenator membrane activates macrophages to produce cytokines. 9 We characterized the SAAcontaining acute phase (AP) HDL during the acute phase to define the polydisperse HDL "substrate" that CETP would encounter. We further investigated CETP function in the acute phase, particularly as it relates to the presence of SAA and sPLA 2 on AP HDL, with respect to its CE transfer and apoA-I liberating functions.Teleologically, the dramatic changes in HDL composition and metabolism during inflammation must serve a short-term purpose to allow the organism to survive a noxious assault. Acute tissue injury results in cell death with large quantities of cell membranes rich in phospholipids and cholesterol generated. Macrophages are mobilized to such sites, ingest these fragments, and acquire considerable lipid load. 10 We thus e...