Low temperature conversion of sludge and shavings from leather industry.

Abstract Brazil has one of the largest herds of cattle in the world, with more than 170 million heads. Over 400 farms have exported more than 2,875 ton (in 1997) of leather to Europe. The wet blue tanning process uses chemicals such as chromium compounds and produces liquid wastes that must be treated by physicochemical and biological systems. About 15,000 ton per month of dewatering sludge with 24% solids content is disposed of into landfills. During the process, pre-tanned skins (wet blue leather) are shaved to the desired thickness and the shavings, like sludge, are among the wastes that must have special attention. The organic content and chromium concentration are high. About 12% of the leather production from cattle hides are shavings, and its chromium concentration ranges from 3.5 to 5.5% of dry matter. The Environmentally friendly leather project, a co-operation between Brazilian and German tanneries, universities and technical schools, is looking for process optimisation, waste minimisation and adequate treatment for solid and liquid wastes from the leather industry. This work presents results of Low Temperature Conversion of chrome-containing sludge and shavings in a laboratory batch reactor, offering a solution for these hazardous wastes, recovering the energy content and transforming metals in insoluble sulphides.

Unsaponifiable lipid constituents of some underutilized tropical seed oils.

Sterols, triterpene alcohols, and hydrocarbons present in the unsaponifiable fraction of some underutilized tropical seed oils have been examined. The seeds include Telfairia occidentalis (TLO), Andenopus breviflorus (ADB), Cucumeropsis edulis (CME), Antiaris africana (ATF), and Monodora tenuifolia (MNT). The oil content of the seeds was high (34.7-68.8%), whereas triacylglycerols comprised the dominant lipid group in the oils (65.4-73.9%). The percentage of unsaponifiables ranged from 1.1 to 7.9%. Ten sterols were identified in the fractions. In the Cucurbitaceae oils (TLO, CME, and ADB), Delta(7)-sterols constituted the dominant sterols. These include 24-ethylcholesta-7,22E,25-trienol (7), 24-ethylcholesta-7,25-dienol (9), 24Z-ethylidenecholes-7-enol (10), and 24-ethylcholesta-7, 24-dienol (11). However Delta(5)-sterols (1-5) occurred at the highest concentration in the other two samples (ATF and MNT). Fifteeen triterpene alcohols were detected in the fractions. Olean-12-enol (16), isomultiflorenol (8), and lupeol (23) were the dominant alcohols in the Cucurbitaceae family, whereas alpha-amyrin (urs-12-enol) (20) was the dominant triterpene alcohol in ATF and MNT. A mixture of C(18)-C(34) n-alkanes, squalene, and some monoterpenes was detected in the hydrocarbon fraction.

Eradication of pre-established lymphoma using herpes simplex virus amplicon vectors.

Herpes simplex virus amplicon vectors expressing RANTES (HSVrantes) and the T-cell costimulatory ligand B7.1 (HSVB7.1) were studied for their ability to elicit a tumor-specific T-cell response in a murine lymphoma model. HSVB7.1- and HSVrantes-transduced EL4 cells expressed high levels of B7.1 and RANTES as analyzed by flow cytometry and enzyme-linked immunosorbent assay, respectively. Inoculation of ex vivo HSVB7.1 transduced cells in syngeneic mice resulted in regression of both transduced cells and nontransduced cells inoculated contralaterally. Direct intratumoral injection of HSVB7.1 and/or HSVrantes alone or in combination into established EL4 tumors led to complete tumor regression in injected tumors as well as in nontransduced contralaterally implanted tumor, whereas control tumors or tumors injected with HSVlac expressing beta-galactosidase did not regress. Maximal protection was achieved with combined injection of HSVB7.1 and HSVrantes; mice showing tumor regression were resistant to rechallenge with parental EL4 cells, and tumor cell-specific cytolytic T-cell activity was observed in mice demonstrating regression. HSV amplicon-mediated delivery of immune effector molecules may represent a useful strategy for immunotherapy in the setting of pre-existing tumor.

Analysis of computer-predicted antibody inducing epitope on Japanese encephalitis virus.

Theoretical methods to delineate antibody inducing epitopes have been employed to predict antigenic determinants on envelope glycoprotein (gpE) of Japanese encephalitis (JE), West Nile (WN) and Dengue (DEN) I-IV viruses. A predicted region on JE virus gpE 74CPTTGEAHNEKRAD87 was synthesized, conjugated to KLH (KLH-peptide) and used in immunization of mice. A mouse monoclonal antibody (MoAb IVB4) reactive to the peptide was also found to react with native JE virus gpE. Characterization of the idiotypic (ID) determinants with the help of polyclonal domain-specific anti-ID antibodies revealed that polyclonal anti-KLH-peptide antibodies and MoAb IVB4 are flavivirus-cross-reactive to Hx and NHx domains, respectively. The region 74-87 in JE virus gpE has been mapped as a linking area between Hx and NHx domains. Reactivity of the peptide with sera from JE patients and vaccinees also indicated the feasibility of using predicted peptides for diagnostic and prophylastic purposes.

Characterization of poliovirus-specific T lymphocytes in the peripheral blood of Sabin-vaccinated humans.

Poliovirus-specific cellular immune responses were identified in the peripheral blood mononucleocytes of Sabin-immunized human donors by using a proliferation assay. Complement depletion and monoclonal antibody inhibition studies suggest that the effector population is the major histocompatibility complex (MHC) class II-restricted CD4+ T-helper cell. Immune lymphocytes proliferated to polyacrylamide gel purified-capsid proteins VP1, VP2, and VP3 and, in some individuals, to synthetic VP4, indicating the presence of T-cell epitopes in each of these proteins. Using synthetic peptides, T-cell epitopes have been mapped to specific regions in VP1 which lie near previously identified neutralizing antibody recognition sites. Human leukocyte antigen (HLA) typing of the donor individuals indicated that no MHC class II molecule was held in common between all four donor individuals. Thus, the positive responses observed with peptides p182-201 and p244-261 in three of four and four of four donors suggest that these peptides contain epitopes presented by at least two different MHC molecules. Antibody-blocking experiments suggest that an epitope within VP1 residues 244 to 264 is presented by HLA DQ3.

Poliovirus-specific major histocompatibility complex class I-restricted cytolytic T-cell epitopes in mice localize to neutralizing antigenic regions.

A major histocompatibility complex (MHC) class I-restricted cytotoxic T-lymphocyte (CTL) response is induced in BALB/c mice upon immunization with poliovirus serotype 1 (Mahoney strain). A similar class I-restricted response is also induced upon immunization with purified VP1 capsid proteins. Thus, poliovirus-specific MHC class I CTL responses can be induced independently of viral infection in murine hosts. In experiments using recombinant vaccinia virus vectors expressing different segments of the poliovirus capsid proteins and synthetic peptides, two regions of the VP1 capsid protein appear to contain epitopes recognized by this bulk CTL population. These epitope regions contain a Kd-restricted peptide-binding motif. Interestingly, each of these CTL epitopes is located near previously defined neutralizing antigenic sites.

Monoclonal antibody to Japanese encephalitis virus cross-reacting with histones present in the cell nuclei.

An immunoglobulin G (IgG2b) class of monoclonal antibody (MoAb, NHA-1) raised against Japanese encephalitis virus (JEV) E glycoprotein, reacted with the viral antigen expressed in cytoplasm of the infected cells and also with the cell nuclei, by an indirect fluorescent antibody technique (FA). The NHA-1 reactivity to nuclei was found to be due to its recognizing a JEV cross-reactive epitope present on the nuclear histones. Adsorption with calf thymus histones (type II-AS) showed a drop in NHA-1 reactivity to both JEV and histones by an enzyme-linked immunosorbent assay (ELISA) and indirect FA; the drop was higher against the histones. The MoAb recognized specifically the viral antigens expressed on the infected porcine kidney cell surface by a modified indirect FA. ELISA carried out with glutaraldehyde-fixed antigens showed an almost 2-fold increase in the reactivity over unfixed JEV antigen but none for the histones. Thus, the results indicate that histones share a sequential homology with E glycoprotein of JEV, which might lead to an autoimmune disorder induced due to the molecular mimicry between these two antigens.

Identification of T-helper epitopes in the VP1 capsid protein of poliovirus.

Poliovirus-specific T lymphocytes were isolated from virus-immunized mice of different H-2 haplotypes. Immunological characterization of this population indicates that the effector population involved in the observed poliovirus-specific proliferative response was that of CD4-positive T-helper cells. Proliferative responses also were induced within these T-lymphocyte populations upon stimulation with either purified VP1 capsid protein or VP1 synthetic peptides. By using these synthetic peptides, several T-helper epitopes were identified. Generally, proliferative responses were observed in three regions of VP1. Two regions spanning VP1 residues 86 to 120 and 201 to 241 were recognized by T lymphocytes from BALB/c (H-2d), C57BL/6 (H-2b), and C3H/HeJ (H-2k) backgrounds. Analyses using synthetic peptides of nonoverlapping sequences indicated that the region spanning residues 201 to 241 may contain several T epitopes and may account for the strong proliferative response observed. In addition, for two of the three haplotypes examined, T epitopes were observed within residues 7 to 24 of VP1. Additional epitopes which appeared to be restricted to specific H-2 backgrounds were identified. T epitopes within VP1 that are common between different strains of mice appeared to lie within previously identified neutralizing antigenic sites in poliovirus.

Characterization of & induction of immune response to anti-idiotypic antibodies for JE virus.

Anti-idiotypic antibodies (anti-Ids, Ab2s) were prepared by immunizing rabbits with two murine monoclonal antibodies (Ab1) having specificities for two independent haemagglutinin (HA) epitopes on JE virus [viz., Hs-1, monoclonal antibody (MAb) specific for Japanese encephalitis virus (JEV) and Hx-1, MAb common to flaviviruses]. Anti-Hs-1 (S-Ab2) and Anti-Hx-1 (X-Ab2) reacted specifically with the immunizing Ab1. In addition, they could react with other MAbs whose reactivity was similar to their immunizing homologous Ab1. The paratope inhibition assay indicated that both anti-idiotypes recognized paratope related idiotopes on their respective Ab1 and could therefore be designated as Ab2 beta. Experimental animals (Swiss mice, Balb/c mice and guineapigs) immunized with S-Ab2 or X-Ab2 produced anti-JE virus antibodies (Ab3) which could be detected by enzyme linked immunosorbent assay, immunofluorescence, haemagglutination inhibition and neutralization tests. The anti-idiotypes were also found to stimulate a cellular immune response in vitro as assessed by 3H thymidine incorporation by lymphocytes from JE vaccinated individuals and experimentally immunized Balb/c mice. The findings of the present study suggest that both the anti-Id antibodies are homobodies which may act as surrogate antigens to manipulate the immune response against JEV.

Recognition of helper T cell epitopes in envelope (E) glycoprotein of Japanese encephalitis, west Nile and Dengue viruses.

Helper T (Th) cell antigenic sites were predicted from the primary amino acid sequence (approximately 500 in length) of the envelope (E) glycoprotein (gp) of Japanese encephalitis (JE), West Nile (WN) and Dengue (DEN) I-IV flaviviruses. Prediction of Th epitopes was done by analyzing the occurrence of amphipathic segments, Rothbard-Taylor tetra/pentamer motifs and presence of alpha helix-preferring amino acids. The simultaneous occurrence of all these parameters in segments of E gp were used as criteria for prediction as Th epitopes. Only one cross reactive epitope was predicted in the C-terminal region of the E gp predicted segments of all flaviviruses analyzed. This region is one of the longest amphipathic stretch (approximately from 420 to 455) and also has a fairly large amphipathic score. Based on the predicted findings three selected peptides were synthesized and analyzed for their ability to induce in vitro T cell proliferative response in different inbred strains of mice (Balb/c, C57BL6, C3H/HeJ). Synthetic peptide I and II prepared from C-terminal region gave a cross reactive response to JE, WN and Den-II in Balb/c and C3H/HeJ mice. Synthetic peptide III prepared from N-terminal region gave a proliferative response to DEN-II in Balb/c strain only, indicating differential antigen presentation.