Virtually all the known physico-chemical and biological techniques have been explored for treatment of extremely recalcitrant dye wastewater; none, however, has emerged as a panacea. A single universally applicable end-of-pipe solution appears to be unrealistic, and combination of appropriate techniques is deemed imperative to devise technically and economically feasible options. An in-depth evaluation of wide range of potential hybrid technologies delineated in literature along with plausible analyses of available cost information has been furnished. In addition to underscoring the indispensability of hybrid technologies, this paper also endorses the inclusion of energy and water reuse plan within the treatment scheme, and accordingly proposes a conceptual hybrid dye wastewater treatment system. Keywords GeoQUEST

We describe here the involvement of calciumactivated neutral protease (CANP or calpain, EC 3.4.22.17) in calcium-dependent proteolytic processing of the precursor of human interleukin la (IL-la) into mature IL-la. Calcium ionophore ionomycin enhanced proteolytic processing of pre-IL-la and the release of mature IL-la either from lipopolysaccharide (LPS)-activated human adherent mononuclear cells or from a human bladder carcinoma cell line (HTB9 5637) that constitutively produces human IL-la and -P. The proteolytic processing of pre-IL-la was completely inhibited by EGTA. Similar calcium-dependent proteolytic processing of pre-IL-la was also observed with lysates of either LPS-activated human adherent mononuclear cells or HTB9 5637 cells. Since the optimal pH for processing was between 7 and 8, and E-64 (a cysteine protease inhibitor) and leupeptin (a serine and cysteine protease inhibitor) both inhibited this processing by cell lysates, we hypothesized that a calcium-activated neutral protease, CANP, might be responsible for this processing. This hypothesis was supported by data showing that the specific CANP inhibitor peptide inhibited this proteolysis in cell lysates in a dose-dependent fashion (IC50 = 0.05 IM) and that treatment of pre-IL-la with purified CANP yielded the 17-kDa mature form of IL-la, which has an amino terminus identical with that reported for mature human IL-la. Taken together, these findings indicate that calcium-dependent proteolytic processing of pre-IL-la is selectively mediated by CANP.Interleukin 1 (IL-1), produced by a variety of cells, is widely known to manifest multiple biological activities on various types of target cells (1). Two biochemically distinct forms of IL-1, IL-la and -/3, have been genetically cloned (2-4). Although both types of IL-1 are lacking in a signal peptide, they have been reported to be present not only in the cytosol (5) but also in the cell membrane (6) and extracellularly. Since mature IL-1 has a molecular mass of 17 kDa, precursor IL-1 (pre-IL-1; molecular mass of 33-35 kDa) presumably is processed by an as-yet-unidentified proteolytic enzyme(s) to generate mature IL-1.A calcium ionophore has been reported to augment production of extracellular IL-1 by lipopolysaccharide (LPS)-stimulated human adherent mononuclear cells through the entry of exogenous calcium into cells (7). LPS also has been shown to increase intracellular calcium in macrophages (8).On the other hand, there are some cell types, such as HTB9 5637, which constitutively produce pre-IL-1 but do not generate and release mature IL-1 effectively (ref. 9; unpublished results). We, therefore, decided to test the possibility that calcium plays some roles in release and/or processing of IL-1 by comparative studies of this cell line and human adherent mononuclear cells. We examined the effects of calcium ionophore on the release and processing of IL-1 in detail, and we identified a major calcium-dependent proteolytic processing enzyme of pre-IL-la as calcium-activated neutral protease (CANP ...

Killer cell lectin-like receptor G1 (KLRG1) is an inhibitory receptor expressed on subsets of natural killer (NK) cells and T cells, for which no endogenous ligands are known. Here, we show that KLRG1 binds three of the classical cadherins (E-, N-, and R-), which are ubiquitously expressed in vertebrates and mediate cell–cell adhesion by homotypic or heterotypic interactions. By expression cloning using the mouse KLRG1 tetramer as a probe, we identified human E-cadherin as a xenogeneic ligand. We also identified a syngeneic interaction between mouse KLRG1 and mouse E-cadherin. Furthermore, we show that KLRG1 binds N- and R-cadherins. Finally, we demonstrate that E-cadherin binding of KLRG1 prevents the lysis of E-cadherin–expressing targets by KLRG1+ NK cells. These results suggest that KLRG1 ligation by E-, N-, or R-cadherins may regulate the cytotoxicity of killer cells to prevent damage to tissues expressing the cadherins.

Methylglyoxal (MG) is one of the side-products in glycolysis, and it reacts with proteins under physiological conditions. Here, we identified heat-shock protein 27 (Hsp27) as a major MG-modified protein in cells. MG modification of Hsp27 selectively occurs at Arg-188 to form argpyrimidine, and mutation in the residue represses the formation of a large oligomer. This modification process is essential to its repressing activity for cytochrome c-mediated caspase activation. Inhibition of MG modification of Hsp27 causes sensitization of the cells to anti-tumor drug-induced apoptosis. Thus, MG is a novel modulator of cell survival by directly incorporating with the specific protein residue.

Lectins on antigen presenting cells are potentially involved in the antigen uptake and the cellular recognition and trafficking. Serial analysis of gene expression in monocyte-derived dendritic cells (DCs), monocytes, and macrophages revealed that 7 of the 19 C-type lectin mRNA were present in immature DCs. Two of these, the macrophage mannose receptor and the macrophage lectin specific for galactose/N-acetylgalactosamine (MGL), were found only in immature DCs, as confirmed by reverse transcriptase-PCR and flow cytometric analysis. By subcloning and sequencing the amplified mRNA, we obtained nucleotide sequences encoding seven different human MGL (hMGL) subtypes, which were apparently derived from alternatively spliced mRNA. In addition, the hMGL gene locus on human chromosome 17p13 contains one gene. A single nucleotide polymorphism was identified at a position in exon 3 that corresponds to the cytoplasmic region proximal to the transmembrane domain. Of all the splicing variants, the hMGL variant 6C was expressed at the highest levels on immature DCs from all donors tested. Immature DCs could incorporate ␣-GalNAc-modified soluble acrylamide polymers, and this was significantly inhibited by pretreatment of the cells with an anti-hMGL monoclonal antibody that blocks the lectin-carbohydrate interaction. We propose that hMGL is a marker of imDCs and that it functions as an endocytic receptor for glycosylated antigens. Dendritic cells (DCs)1 play a pivotal role in the immune system by processing and presenting a variety of antigens to T cells (1). The uptake of exogenous antigens is the first step in this process and is therefore a critical event that influences DC function. The uptake of glycoconjugates by DCs is potentially mediated by lectins, which are carbohydrate-binding proteins. There are at least four distinct lectin families in animal cells known as the C-type, S-type, P-type, and I-type lectins (2). Some lectins are known to participate in molecular and cellular trafficking in a manner that is dependent on the lectin type, its molecular architecture, and its subcellular localization. DCs are known to express a variety of lectins, particularly C-type lectins, but as yet their biological roles in DC function are unclear.How glycosylated antigen presentation is regulated and how this affects the subsequent immune responses has not yet been clarified. This is an important issue to investigate as it may improve our understanding of, for example, anti-tumor immunity to MUC1. MUC1 is a glycosylated membrane protein that frequently expresses truncated O-glycans such as the T (Gal␤1-3GalNAc-Thr/Ser) and Tn (GalNAc-Thr/Ser) antigens. MUC1 is an important candidate vaccine antigen as it is an antigen that is often overexpressed in solid tumors, including carcinoma of the breast, lung, pancreas, colon, and ovaries. MUC1-specific cytotoxic T lymphocytes have been isolated from draining lymph nodes of pancreatic and breast cancer patients, ascitic fluids of ovarian cancer patients, and peripheral blood mononuclear cell...

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Growing evidence indicates that N-glycans function as tags recognized by a variety of intracellular lectins that govern the fates of glycoproteins in eukaryotes (1-4). These lectins are postulated to interact specifically with partially trimmed intermediates of high-mannose-type oligosaccharides displayed on the target polypeptide and thereby to control protein translocation, folding, degradation, and trafficking.The lectins calreticulin and calnexin assist in the folding process of nascent proteins in the endoplasmic reticulum (ER) 5 by capturing their monoglucosylated high-mannosetype oligosaccharides after the action of glucosidase I (1-6). Glucosidase II then removes the remaining glucose residue at the non-reducing terminal of the D1 branch of the substrate glycoproteins, giving rise to the M9 glycoform (the oligosaccharides are designated as shown in Fig.

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