Recurrent membranous nephropathy and acute cellular rejection in a patient treated with direct anti-HCV therapy (ledipasvir/sofosbuvir).

Direct-acting antiviral agents (DAAs) are very effective therapy for chronic hepatitis C infection, and have revolutionized the treatment of hepatitis C in kidney allograft recipients. Although well tolerated in general, rare renal complications have been reported. We describe a case of recurrent membranous nephropathy and acute cellular rejection in a kidney allograft recipient after DAA (ledipasvir/sofosbuvir) therapy, whose allograft function had been stable for more than 30 years. The patient was presented with nephrotic range proteinuria with stable creatinine. The kidney allograft biopsy revealed recurrent membranous nephropathy with fine granular deposits of IgG1/IgG4 codominance and positive phospholipase A2 receptor (PLA2R) staining. The patient was treated with pulse steroid and rituximab, leading to a decrease in proteinuria. As DAAs are more frequently used, physicians should be aware of immune-related renal complications.

Determination of fluoranthene by antigen-coated indirect competitive real-time immuno-PCR assay.

A reliable and sensitive antigen-coated indirect competitive real-time immuno-PCR (RT-IPCR) assay was developed for the determination of fluoranthene (FL). 9-Fluoranthenebutanoic acid, gamma-oxo-(FLA) was synthesized as the hapten for FL. Mixed anhydride reaction (MAR) was used to couple the FLA to ovalbumin (OVA) to form artificial coating antigen. Active ester method (AEM) was used to couple the FLA to bovine serum albumin (BSA) to form artificial immune antigen. Male New Zealand white rabbits were immunized with immune antigen to obtain polyclonal antibodies (pAbs), with which, a novel RT-IPCR assay for determination of FL was described. Under best conditions, FL can be determined in a concentration range between 10 fg mL(-1) and 100 pg mL(-1) with a detection limit of 5 fg mL(-1). The cross-reactivities of the anti-FL antibody to seven structurally related compounds were below 11%. Some environmental water samples were analyzed with satisfactory results, which shows a good accuracy and suitability to analyze FL in environmental water. Recovery was lower or higher with agitation but would still be acceptable for use in an on-site field test to provide rapid, semi-quantitative, and reliable test results for making environmental decisions.

Isolation of priority polycyclic aromatic hydrocarbons from natural sediments and sludge reference materials by an anti-fluorene immunosorbent followed by liquid chromatography and diode array detection.

The selective isolation of PAHs from complex environmental mixtures was accomplished by means of a new methodology based on antigen--antibody interactions. This method consists on the extraction of PAHs from water samples onto an anti-fluorene immunosorbent (IS) followed by liquid chromatography with diode array detection. Environmental sediments and a sludge reference material containing PAHs were analyzed using this methodology in order to validate the performance of the IS for the cleanup procedure of such materials. Sediments were extracted by sonication with dichloromethane/methanol (2:1), and the extracts were brought to a volume of 100 mL of water in order to perform the extraction with the anti-fluorene IS. The reliability of the cleanup achieved by the IS was well demonstrated in the analysis of sediment and sludge complex samples containing the priority PAHs established by the U.S. EPA at concentrations varying from 56 micrograms/kg to 26 mg/kg. Results were compared to those obtained with conventional cleanup procedures showing a better selectivity for PAHs. The chromatograms presented a clear baseline allowing the determination and quantification of PAHs at the ppb level. Using immunosorbents, extraction, trace enrichment, and cleanup were accomplished in only one step.

A rapid and sensitive screening method for the detection of anti-2-acetylaminofluorene immunoglobulins.

A method is described in which anti-2-acetylaminofluorene immunoglobulins may be detected using a simple and sensitive screening procedure. The method is based on immunoglobulin binding of an 125I derivatized 2-aminofluorene radiotracer. Tracer binding is not isotype specific, and thus the method is useful for the detection of either IgG or IgA. Competitive binding experiments with the radiotracer were used to determine the specificity of immunoglobulin response by measurement of cross-reactivity with related ligands. This method allows quantitation of the immune response to the carcinogen in serum and other biological fluids (i.e., intestinal secretions).

Biocompatibility studies with a novel perfluorochemical emulsion.

Biocompatibility studies with a novel perfluorochemical (PFC) emulsion have been performed in rats. Injection of the emulsion was followed by changes in lymphoid tissue weights, comparable but not identical to those produced by the commercial formulation, Fluosol-DA 20% (F-DA). Peritoneal injection of either the novel emulsion or F-DA enhanced the humoral immune response to intraperitoneally-injected sheep red blood cells (SRBC) suggesting immunopotentiating properties of emulsion components.

Antibodies to N-(guanosin-8-yl)-2-aminofluorene.

Antibodies to Guo-AF were elicited in rabbits by immunization with bovine serum albumin -Guo-AF conjugate. The antibodies were purified by affinity chromatography. The affinity of the antibodies towards several ligands was studied by radioimmunoassays. The antibodies recognize Gua-AAF and Guo-AF (K(Guo-AF)/K(Guo-AAF) congruent to 25). The circular dichroism of Guo-AF as a function of pH is reported.

Antibodies to N-hydroxy-2-aminofluorene modified DNA as probes in the study of DNA reacted with derivatives of 2-acetylaminofluorene.

Antibodies were elicited in rabbits immunized with a mixture of methylated bovine serum albumin and N-hydroxy-2-aminofluorene reacted DNA (DNA-AF). These antibodies were used in competition radioimmunoassays (RIA) with [3H]DNA-AF as radioactive antigen and different N-acetoxy-2-acetylaminofluorene (N-AcO-AAF) and N-hydroxy-2aminofluorene (N-OH-AF) reacted DNAs, deoxyguanosine, dGMP, dGpA and dApG as inhibitors. Based on the results of RIA it is concluded that the binding sites of the two residues, -AF and -AAF, to guanosine in DNA, are essentially the same. Moreover, the same sites seem to be reactive also in the direct reaction of N-AcO-AAF with dGMP. Circular dichroism measurements of modified DNA confirm the RIA results.