African-American recipients of kidney transplants with lupus have high allograft failure risk. We studied their risk adjusting for: (1) socio-demographic factors: donor age, gender and race-ethnicity; recipient age, gender, education and insurance; donor–recipient race-ethnicity match; (2) immunologic factors: donor type, panel reactive antibodies, HLA mismatch, ABO blood type compatibility, pre-transplant dialysis, cytomegalovirus risk and delayed graft function (DGF); (3) rejection and recurrent lupus nephritis (RLN). Two thousand four hundred and six African-, 1132 Hispanic-, and 2878 Caucasian-Americans were followed for 12 years after transplantation. African- versus Hispanic- and Caucasian-Americans received more kidneys from deceased donors (71.6%, 57.3% and 55.1%) with higher two HLA loci mismatches for HLA-A (50%, 39.6% and 32.4%), HLA-B (52%, 42.8% and 35.6%) and HLA-DR (30%, 24.5% and 21.1%). They developed more DGF (19.5%, 13.6% and 13.4%). More African- versus Hispanic- and Caucasian-Americans developed rejection (41.7%, 27.6% and 35.9%) and RLN (3.2, 1.8 and 1.8%). 852 African-, 265 Hispanic-, and 747 Caucasian-Americans had allograft failure ( p < 0.0001). After adjusting for transplant era, socio-demographic-immunologic differences, rejection and RLN, the increased hazard ratio for allograft failure of African- compared with Caucasian-Americans became non-significant (1.26 [95% confidence interval 0.78–2.04]). African-Americans with lupus have high prevalence of risk factors for allograft failure that can explain poor outcomes.

Systemic lupus erythematosus may present with renal manifestations that frequently are difficult to categorize and lupus nephritis is an important predictor of poor outcome. The type and spectrum of renal injury may remain undiagnosed until full-blown nephritic and/or nephrotic syndrome appear with increased risk of end-stage renal disease. These abnormalities occur within the first few years after the diagnosis of lupus is made on clinical grounds and with the support of laboratory tests in high risk patients. An early renal biopsy is helpful in patients with an abnormal urinalysis and/or reduced glomerular filtration rate and the results form the basis for therapeutic decisions. The biopsy also provides vital prognostic information based on histological categorization of different types of lupus nephritis, the degree of activity, chronicity and the immunopathogenesis. In the current armamentarium, the use of cyclophosphamide and azathioprine and recently mycophenolate mofetil, reduce morbidity and maintenance therapies reduce the risk of end-stage renal disease. Clinical trials underway promise new, effective and safe immunosuppressive regimens for the treatment of proliferative lupus nephritis.

Procedures for sampling genomic DNA from live billfishes involve manual restraint and tissue excision that can be difficult to carry out and may produce stresses that affect fish survival. We examined the collection of surface mucous as a less invasive alternative method for sourcing genomic DNA by comparing it to autologous muscle tissue samples from Atlantic blue marlin (Makaira nigricans), white marlin (Tetrapturus albidus), sailfish (Istiophorus platypterus), and swordfish (Xiphias gladius). Purified DNA from mucous was comparable to muscle and was suitable for conventional polymerase chain reaction, random amplified polymorphic DNA analysis, and mitochondrial and nuclear locus sequencing. The nondestructive and less invasive characteristics of surface mucous collection may promote increased survival of released specimens and may be advantageous for other marine fish genetic studies, particularly those involving large live specimens destined for release.

Journal of Applied IchthyologyVolume 23, Issue 2 p. 184-188 Tracking gender factors in fish surface mucus: temporal patterns in individual Koi (Cyprinus carpio) D. R. Schultz, D. R. Schultz Department of Medicine, University of Miami School of MedicineSearch for more papers by this authorN. Perez, N. Perez Department of Medicine, University of Miami School of MedicineSearch for more papers by this authorA. J. Mendez, A. J. Mendez Clinical Chemistry Laboratory, Diabetes Research Institute, University of Miami School of MedicineSearch for more papers by this authorD. Snodgrass, D. Snodgrass Southeast Fisheries Science Center, National Marine Fisheries Service, Miami, FLSearch for more papers by this authorJ. E. Serafy, J. E. Serafy Southeast Fisheries Science Center, National Marine Fisheries Service, Miami, FLSearch for more papers by this authorE. D. Prince, E. D. Prince Southeast Fisheries Science Center, National Marine Fisheries Service, Miami, FLSearch for more papers by this authorW. A. Crow Jr, W. A. Crow Jr Department of Medicine, University of Miami School of MedicineSearch for more papers by this authorT. R. Capo, T. R. Capo Rosenstiel School of Marine and Atmospheric Science, University of Miami, Miami, FL, USASearch for more papers by this author D. R. Schultz, D. R. Schultz Department of Medicine, University of Miami School of MedicineSearch for more papers by this authorN. Perez, N. Perez Department of Medicine, University of Miami School of MedicineSearch for more papers by this authorA. J. Mendez, A. J. Mendez Clinical Chemistry Laboratory, Diabetes Research Institute, University of Miami School of MedicineSearch for more papers by this authorD. Snodgrass, D. Snodgrass Southeast Fisheries Science Center, National Marine Fisheries Service, Miami, FLSearch for more papers by this authorJ. E. Serafy, J. E. Serafy Southeast Fisheries Science Center, National Marine Fisheries Service, Miami, FLSearch for more papers by this authorE. D. Prince, E. D. Prince Southeast Fisheries Science Center, National Marine Fisheries Service, Miami, FLSearch for more papers by this authorW. A. Crow Jr, W. A. Crow Jr Department of Medicine, University of Miami School of MedicineSearch for more papers by this authorT. R. Capo, T. R. Capo Rosenstiel School of Marine and Atmospheric Science, University of Miami, Miami, FL, USASearch for more papers by this author First published: 31 January 2007 https://doi.org/10.1111/j.1439-0426.2006.00799.xCitations: 7 Author's address: D. R. Schultz, Department of Medicine, University of Miami School of Medicine, R-102, PO Box 016960, Miami, FL 33101, USA.E-mail: [email protected] Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL References Barbieri, L. R.; Chittenden, M. E. Jr; Lowerre-Barbieri, S. K., 1994: Maturity, spawning, and ovarian cycle of Atlantic croaker, Micropogonias undulatus, in the Chesapeake Bay and adjacent coastal waters. US Fish. Bull. 92, 671– 685. Barry, T. P.; Santos, A. J. G.; Furukawa, K.; Aida, K.; Hanyu, I., 1990: Steroid profiles during spawning in male common carp. Gen. Comp. Endocrinol. 80, 223– 231. Borg, B., 1994: Androgens in teleost fishes. Comp. Biochem. Physiol. C 109, 219– 245. Chang, C.-F.; Chen, M.-R., 1990: Fluctuation in sex steroids and sex steroid-binding protein during the development and annual cycle of the male common carp, Cyprinus carpio. Comp. Biochem. Physiol. 97A, 565– 568. Copeland, P. A.; Thomas, P., 1988: The measurement of plasma vitellogenin levels in a marine teleost, the spotted seatrout (Cynoscion nebulosus) by homologous radioimmunoassay. Comp. Biochem. Physiol. 91B, 17– 23. Cuisset, B.; Pradelles, P.; Kime, D. E.; Kühn, E. R.; Babin, P.; Davail, S.; Le Menn, F., 1994: Enzyme immunoassay for 11-ketotestosterone using acetylcholinesterase as label: application to the measurement of 11-ketotestosterone in plasma of Siberian sturgeon. Comp. Biochem. Physiol. 108C, 229– 241. Cuisset, B.; Fostier, A.; Williot, P.; Bennetau-Pelissero, C.; Le Menn, F., 1995: Occurrence and in vitro biosynthesis of 11-ketotestosterone in Siberian sturgeon, Acipenser baeri Brandt maturing females. Fish Physiol. Biochem. 14, 313– 322. Cyr, D. G.; Idler, D. R.; Audet, J. M.; Mcleese, J. M.; Eales, J. G., 1998: Effects of long-term temperature acclimation on thyroid hormone deiodinase function, plasma thyroid hormone levels, growth, and reproductive status of male Athlantic cod, Gadus morhua. Gen. Comp. Endrocrinol. 109, 24– 36. Ding, J. L.; Hee, P. L.; Lam, T. J., 1989: Two forms of vitellogenin in the plasma and gonads of male tilapia (Oreochromis aureus). Comp. Biochem. Physiol. 93B, 363– 370. Goodbred, S. L.; Gilliom, R. J.; Gross, T. S.; Denslow, N. P.; Bryant, W. L.; Schoeb, T. R., 1996: Reconnaissance of 17 ß-estradiol, 11-ketotestosterone, vitellogenin, and gonad histopathology in common carp of United States Streams: potential for contaminant-induced endocrine disruption. USGS-Pesticide National Synthesis Project Pesticides Report. US Geological Survey Open-File Report 96–627, USGS, Denver, CO. Goodwin, A. E.; Grizzle, J. M.; Bradley, J. T.; Estridge, B. H., 1992: Monoclonal antibody-based immunoassay of vitellogenin in the blood of male channel catfish (Ictalurus punctatus). Comp. Biochem. Physiol. 101B, 441– 446. Gordon, M. P.; Owen, T. G.; Ternan, T. A.; Hildbrand, L. K., 1984: Measurement of a sex-specific protein in skin mucus of premature coho salmon (Oncorynchus kisutch). Aquaculture 43, 333– 339. Grier, H. J., 2002: The germinal epithelium: its dual role in establishing male reproductive classes and understanding the basis for indeterminate egg production in female fishes. Proc. Gulf Carib. Fish. Inst. 53, 537– 552. Kime, D. E.; Nash, J. P.; Scott, A. P., 1999: Vitellogenesis as a biomarker of reproductive disruption by xenobiotics. Aquaculture 177, 345– 352. Kishida, M.; Anderson, T. R.; Specker, J. L., 1992: Induction of vitellogenin in striped bass (Morone saxatilis): characterization and quantification in plasma and mucus. Gen. Comp. Endocrinol. 88, 29– 39. Laemmi, U. K., 1970: Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 227, 680– 685. Leatherland, J. F.; Copeland, P.; Sumpter, J. P.; Sonstegard, R. A., 1982: Hormonal control of gonadal maturation and development of secondary sexual characteristics in coho salmon, Oncorhynchus kisutch, from Lakes Ontario, Erie and Michigan. Gen. Comp. Endocrinol. 48, 196– 204. Lokman, P. M.; Vermeulen, G. J.; Lambert, J. G. D.; Young, G., 1998: Gonad histology and plasma steroid profiles in wild New Zealand freshwater eels (Anguilla dieffenbachia and A. australis) before and after onset of the natural spawning migration. I. Females. Fish Physiol. Biochem. 19, 325– 338. Lowerre-Barbieri, S. K.; Chittenden, M. E., Jr; Barbieri, L. R., 1996: The multiple spawning patterns of weakfish, Cynoscion regalis, in the Chesapeake Bay and Middle Atlantic Bight. J. Fish. Biol. 48, 1139– 1163. Mommsen, P. T.; Walsh, P., 1988: Vitellogenesis and oocyte assembly. In: Fish physiology, Vol. 11A. W. S. Hoar, D. J. Randall and E. M. Donaldson (Eds). Academic Press, New York, pp. 347– 406. Moncaut, N.; Nostro, F.; Maggese, M. C., 2003: Vitellogenin detection in surface mucus of the South American cichlid fish Cichlasoma dimerus (Heckel, 1840) induced by estradiol-17ß. Effects on liver and gonads. Aquat. Toxicol. 63, 127– 137. Schultz, D. R.; Perez, N.; Tan, C.-K.; Mendez, A. J.; Capo, T. R.; Snodgrass, D.; Prince, E. D.; Serafy, J. E., 2005: Concurrent levels of 11-ketotestosterone in fish surface mucus, muscle tissue and blood. J. Appl. Ichthyol. 21, 394– 398. Slater, C. H.; Schreck, C. B.; Swanson, P., 1994: Plasma profiles of the sex steroids and gonadotropins in maturing female spring Chinook salmon (Oncorhynchus tshawytscha). Comp. Biochem. Physiol. 109A, 167– 175. Sumpter, J. P., 1991: The purification, radioimmunoassay and plasma levels of vitellogenin from the rainbow trout, Salmo gairdneri. In: Current trends in comparative endocrinology, Vol. 1. B. Lofts and W. N. Holmes (Eds). Hong Kong University Press, Hong Kong, pp., 355– 357. Tan, C.-K.; Perez, N.; Mendez, A. J.; Snodgrass, D.; Prince, E. D.; Serafy, J. E.; Arocha, F.; Schultz, D. R., 2006: Identification of vitellogenic Atlantic blue marlin (Makaira nigricans) from muscle samples using an ELISA for vitellogenin-derived yolk proteins. Bull. Mar. Sci., 78, 319– 329. Towbin, H.; Staelin, T.; Gordon, J., 1979: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl Acad. Sci. USA 76, 4350– 4354. Wiegand, M. D., 1996: Composition, accumulation and utilization of yolk lipids in teleost fish. Rev. Fish Biol. Fish. 6, 259– 286. Citing Literature Volume23, Issue2April 2007Pages 184-188 ReferencesRelatedInformation

The purpose of this study was to develop a direct sandwich enzyme-linked immunosorbent assay (ELISA) for vitellogenin-derived yolk proteins (VDYP) in muscle tissue of pelagic Atlantic blue marlin. Gravid ovaries were extracted with physiologic buffer, and purification steps by chromatography were monitored with a cross-reactive mouse monoclonal antibody produced to swordfish (Xiphias gladius Linnaeus, 1758) VDYP. A polyclonal goat antiserum was produced to the enriched VDYP from which specific IgG was purified by affinity chromatography, using immobilized VDYP as the immunoadsorbent. A direct sandwich ELISA was developed using the purified IgG for both capture and detection of the VDYP in standard samples up to a concentration of 1000 ng/ml, and in physiologic buffer-extracted muscle tissue of test samples, using biotin-streptavidin-peroxidase technology. The assay distinguished vitellogenic from non-vitellogenic females and males in extracts of muscle tissue from 27 specimens collected from different areas of the western North Atlantic Ocean during May-September, 2002-2003. The assay for VDYP, and a second for the fish androgen 11-ketotestosterone are in use to indicate gender of electronically-tagged blue marlin from biopsy samples taken just before fish release.

A rapid, reproducible method is described for extracting and comparing levels of the ether-soluble fish androgen 11-ketotestosterone (11-KT) in blood serum, muscle tissue and surface mucus. Because widely different volumes of extract were recovered after centrifugation from the three sources, it was important to express androgen levels as pg 11-KT/mg of total soluble protein (TSP). For six male and four female sexually-staged freshwater Koi (Cyprinus carpio), the method yielded similar pg 11-KT/mg TSP ratios in blood serum and extracts of muscle tissue and surface mucus, with the strongest correlation between blood serum and surface mucus. While male Koi were distinguishable from females based on the magnitude of 11-KT levels, reproductive stage and gonadosomatic index levels were not correlated with the 11-KT levels of either sex. Similar pg 11-KT/mg TSP ratios were also found for autologous muscle tissue and surface mucus extracts of 37 captured and sexed wild marine fishes representing seven genera. However, high 11-KT levels were not restricted to mature males. Collectively, results suggest that surface mucus collection (followed by 11-KT assay) is a useful alternative to more invasive methods of determining systemic hormone levels in fish. Without knowledge of seasonal variation in levels of this and other sex hormones, however, reliance on 11-KT levels alone may lead to spurious identification of gender, let alone reproductive stage.

OBJECTIVES:To characterize cell surface receptors, their ligands, and their proteins in the 2 major pathways of apoptosis; the components that promote/suppress these interactions; the noninflammatory removal of apoptotic bodies by dendritic cells; and methods of assay in studies of cell death. To describe: how deregulation of apoptosis may contribute to autoimmunity, cancer, and neurodegenerative disorders and strategies some viruses have evolved that interfere with the host's apoptotic pathways.METHODS:The authors reviewed and compiled literature on the extrinsic (tumor necrosis factor [TNF] receptor superfamily and ligands) and intrinsic (mitochondria-associated) apoptotic pathways, the pro- and antiapoptotic proteins of the B-cell follicular lymphoma (Bcl)-2 family, the nuclear factor (NF)-kappaB family of proteins, commonly used laboratory methods to distinguish apoptosis from necrosis, the recognition and removal by phagocytosis of apoptotic cells by dendritic cells, and viral strategies to avoid a host's apoptotic response.RESULTS:The 2 major pathways of apoptosis are (1). FasL and other TNF superfamily ligands induce trimerization of cell-surface death receptors and (2). perturbated mitochondria release cytochrome c, the flavoprotein apoptosis-inducing factor, and second mitochondria-derived activator of caspases/DIABLO (a protein that directly neutralizes inhibitors of apoptotic proteins and activates proteases). Catalytically inactive cysteine proteases, called caspases, and other proteases are activated, ultimately leading to cell death with characteristic cellular chromatin condensation and DNA cleavage to fragments of approximately 180 bp. The inhibitory/promoting action of Bcl-2 family members is involved in the release of cytochrome c, an essential factor for the mitochondrial-associated pathway. A balance between inhibition/promotion determines a cell's fate. The NF-kappaB family in the cytoplasm of cells activates various genes carrying the NF-kappaB response element, such as members of the inhibitor of apoptotic proteins family. A few of the more common methods to detect apoptotic cell death are described, which use immunochemical, morphologic and flow cytometric methods, and genetic markers. Exposed phosphatidylserine at the outer leaflet of the plasma membrane of the apoptotic cell serves as a possible receptor for phagocytosis by immature dendritic cells. These cells phagocytize both apoptotic and necrotic cells, but only the latter induce maturation to become fully functional antigen-presenting cells. Viral inhibitors of apoptosis allow increased virus replication in cells, possibly resulting in their oncogenicity.CONCLUSIONS:Balanced apoptosis is crucial in development and homeostasis, and all multicellular organisms have a physiologically programmed continuum of pathways to apoptotic cell death. Further studies of the control at the molecular level of key components and promoters/suppressors of apoptosis may provide better approaches to treatment of autoimmune diseases, malignancies, and neurodegenerative disorders. Many important questions remain regarding the advantages of modifying apoptotic programs in clinical situations.

OBJECTIVEThe complement component C1s is present in dog joint fluid in an activated state. Since C1s degrades insulin-like growth factor binding protein 5 (IGFBP-5), we undertook to determine whether inhibiting C1s in joint fluid would result in an increase in the amount of intact IGFBP-5 and IGF-1 in cartilage and joint fluid, and whether C1s inhibition would be associated with a reduction in cartilage destruction during the development of osteoarthritis (OA).METHODSTwenty-two dogs were randomized to 3 treatment groups. All dogs underwent anterior cruciate ligament transection and were exercised. Dogs received 1 of 3 treatments: buffer alone (controls; n = 6); PB-145, a peptide derived from the sequence of antithrombin III (n = 9); and pentosan polysulfate (PPS; n = 7). PB-145 or saline was injected into the joint space 3 times per week for 3 weeks. PPS was injected intramuscularly weekly for 3 weeks.RESULTSJoint histology showed preservation of chondrocytes and a smooth joint surface in the animals treated with PB-145 and PPS. Mankin scoring showed statistically significant reductions in joint destruction with PB-145 and PPS treatments (P < 0.01) compared with buffer control. Mean active collagenase concentrations were decreased by these two treatments. Immunoblotting of joint fluid showed that both treatments increased concentrations of intact IGFBP-5. Direct analysis of IGFBP-3 and IGFBP-5 protease activity showed that IGFBP-5 was degraded more rapidly and that PB-145 and PPS inhibited the degradation of both proteins. Total IGF-1 concentrations in joint fluid were increased 5.6-5.8-fold by these two treatments. Analysis showed that C1s was being activated in joint fluid and that its activation was inhibited by the addition of PB-145 or PPS.CONCLUSIONThe findings suggest that direct inhibition of the serine protease C1s results in increased concentrations of intact IGFBP-5 and that proteolysis of IGFBP-3 is also inhibited, probably by the inhibition of some other protease. This increase in concentrations of intact IGFBP-3 and IGFBP-5 leads to an increase in IGF-1 which is associated with an improvement in joint architecture during the development of OA.

Capping and release of membranous, small (< 1.5 microm) endothelial microparticles were quantified by immunofluorescence microscopy and flow cytometry after treatment of cultures of human renal microvascular endothelial cells with agonists tumor necrosis factor-alpha (TNF-alpha) or mitomycin C. For constitutive marker CD31, both agonist-treated attached, monolayer, and detached, free endothelial cells formed caps and released microparticles. TNF-alpha and mitomycin C induced dissimilar appearing CD31-containing caps after 3 h, followed by endothelial microparticle release after 6 h. The degree of capping correlated with increasing counts of released microparticles. For lymphokine-inducible CD54, TNF-alpha also induced CD54-containing caps and microparticle release, but mitomycin C failed to induce the expression of either entity. Neither capping nor microparticle release caused by TNF-alpha was part of an apoptotic pathway that involved caspase 3. Mitomycin C treatment of endothelial cells caused capping and microparticle release with a time course similar to TNF-alpha induction for 15 to 24 h, but assays for caspase 3 were positive, confirming the apoptotic action of mitomycin C. Membrane capping and microparticle release from endothelial cells are a convenient experimental model for studying protein movement, release of microparticles, and their possible biological significance.

HomeArteriosclerosis, Thrombosis, and Vascular BiologyVol. 22, No. 10Acquired HDL Deficiency Associated With Apolipoprotein A-I Reactive Monoclonal Immunoglobulins Free AccessLetterPDF/EPUBAboutView PDFView EPUBSections ToolsAdd to favoritesDownload citationsTrack citationsPermissions ShareShare onFacebookTwitterLinked InMendeleyReddit Jump toFree AccessLetterPDF/EPUBAcquired HDL Deficiency Associated With Apolipoprotein A-I Reactive Monoclonal Immunoglobulins Armando J. Mendez, Ronald B. Goldberg, Patricia I. Arnold and Duane R. Schultz Armando J. MendezArmando J. Mendez Department of Medicine, University of Miami School of Medicine, Florida , Ronald B. GoldbergRonald B. Goldberg Department of Medicine, University of Miami School of Medicine, Florida , Patricia I. ArnoldPatricia I. Arnold Department of Medicine, University of Miami School of Medicine, Florida and Duane R. SchultzDuane R. Schultz Department of Medicine, University of Miami School of Medicine, Florida Originally published1 Oct 2002https://doi.org/10.1161/01.ATV.0000032153.19645.5CArteriosclerosis, Thrombosis, and Vascular Biology. 2002;22:1740–1741To the Editor:Immunoglobulins (Igs) directed at components of lipoproteins to cause altered lipoprotein metabolism have been described in patients with multiple myeloma,1,2 xanthomatosis,3 benign gammopathies,4 rheumatoid arthritis,5 systemic lupus erythematosus (SLE), and primary antiphospholipid syndrome (APS).6,7 In most cases, hyperlipidemia results from Igs reactive with apolipoprotein (apo) B present on very low–density lipoproteins and LDLs. Less common have been associations of Igs with hypolipidemia or reactive with HDLs. Here we describe two patients who developed low HDL-cholesterol (HDL-C) levels (<5th percentile) associated with benign gammopathy and the presence of serum Igs reactive with apo A-I.Patient 1 (P1) was a morbidly obese (body mass index, 48 kg/m2), 45-year-old woman with mild normocytic, normochromic anemia (hematocrit, 33 g/L; hemoglobin, 11.6 g/L) and low HDL-C (0.22 mmol/L [8 mg/dL]). Serum Ig levels were elevated. Antinuclear antibodies were detected but were nonreactive with dsDNA, Sm, RNP, SSA, SSB, SCL-70, and histone. Anticardiolipin and anti–β2-glycoprotein I antibodies were negative. Immunofixation electrophoresis (IFE) identified monoclonal IgGκ and IgGλ bands. No Igs or light chains were detected in her urine.Serum lipid levels showed that HDL-C was in the low-normal to normal range between 1992 and 1996 (1.0 to 1.4 mmol/L) but at the time of presentation (November 1999), HDL was markedly reduced (0.22 mmol/L). Concomitantly, there was an increase in serum triglyceride (from <1.3 to >2.5 mmol/L) and a decrease in LDL-C levels (from >3.8 to <2.2 mmol/L). Clinical features were fatigue and the development of asthma. In July 2000, a right perinephric mass was identified as a B-cell lymphoma. High-dose prednisone therapy was initiated and completed in January 2001. The anemia and asthma improved, the perinephric mass disappeared, IgA and IgM levels returned to normal, and IgG, IgGκ, and IgGλ were reduced to subnormal levels. Treatment increased HDL-C (1.14 mmol/L [44 mg/dL]), and the patient is currently well.A second patient (P2) was later identified with an apparent acquired HDL deficiency. This healthy 61-year-old woman was receiving estrogen replacement but had a mild normocytic anemia (hematocrit 33 g/L). Serum lipid results dating to 1987 showed a gradual decline in HDL-C from 1.32 to 0.26 mmol/L (51 to 10 mg/dL), concomitant with increasing triglycerides (from 0.9 to 2.0 mmol). Ig levels were within the normal range, and serum was negative for antinuclear and anticardiolipin antibodies. IFE revealed a monoclonal IgGλ band. The patient remains well apart from the mild anemia and the recent development of peripheral neuropathy.Neither patient was taking lipid-modifying medications, had metabolic abnormalities affecting HDL (such as diabetes, liver or renal disease), or had clinically defined SLE or APS.We tested the hypothesis that the patients were producing antibodies reactive with HDL. Patient and normolipidemic control sera were incubated with goat anti–apo A-I to precipitate HDL, and the washed, solubilized immunoprecipitates were tested for the presence of human Igs by Ouchterlony immunodiffusion. For P1, precipitin bands formed with antibodies to IgG, κ, and λ light chains (Figure 1A), the same IgG subtypes identified by IFE. IgM and IgA were not detected in the immunoprecipitates (not shown). With P2, the solubilized immunoprecipitate reacted only with antibodies to IgG and λ chains (Figure 1B), also identifying the same Ig subtype observed by IFE. Control sera were negative. These data demonstrated the association of Ig with HDL in the serum of the two patients. Download figureDownload PowerPointFigure 1. P1 (A) and P2 (B) serum were incubated with goat anti–apo A-I, and the resulting immunoprecipitates were analyzed by using Ouchterlony immunodiffusion. Wells A and B, solubilized immunoprecipitate; wells 1, 2, and 3, anti-human IgG, anti-κ chain, and anti-λ chain, respectively. C, Apo A-I separated by SDS-PAGE and transferred to nitrocellulose was immunoblotted with 1 μg/mL purified IgG from P1 serum or with 0.5 μg/mL goat anti–human apo A-I IgG as indicated. Arrow indicates migration of apo A-I.Next, apo A-I was subjected to electrophoresis, and after transfer to nitrocellulose, individual lanes were incubated with purified P1 IgG or control goat anti–apo A-I (Figure 1C). P1 IgG, similar to goat anti–apo A-I, reacted specifically with apo A-I. Similar results were obtained with serum IgG isolated from P2 although the reaction was less intense (not shown).The presence of apo A-I–reactive antibodies was associated with significant abnormalities of HDL composition in the patients' serum reflected by the near absence of larger, mature HDL2, and lipid analysis revealed relatively lipid-deficient HDL. Apo A-I–reactive Igs might induce low HDL levels if antibodies diminished the ability of nascent HDL to become lipidated and form mature particles8 or if immune complexes containing apo A-I or HDL were rapidly cleared by the reticuloendothelial system. We found that immune complexes formed between apo A-I and both patients' IgG in vitro induced apo A-I degradation by cultured macrophages. Thus, clearance of immune complexes in vivo could induce catabolism of apoA-I, potentially preventing HDL maturation and leading to reduced HDL levels.Apo A-I can be autoimmunogenic in humans in vivo. Apo A-I–reactive antibodies have been described in subjects with SLE and APS, although a relationship between the antibodies and HDL levels was not directly reported.7 Neither patient had clinically defined SLE or APS. Levy et al9 reported low HDL-C in patients with benign monoclonal gammopathy, multiple myeloma and Waldenstrom's macroglobulinemia. More than half of 49 patients showed an extra lipid band on lipoprotein electrophoresis, suggesting Ig-lipoprotein complexes, but apo A-I and apo B were not detectable in the extra lipid-staining band.Additional mechanisms could account for low HDL. Acute inflammation causes lipoprotein abnormalities mediated by acute phase reactants and (or) cytokines (reviewed by Khovidhunkit et al10). Inflammatory mediators may have influenced HDL concentrations in our patients, particularly because raised CRP levels were identified in P1; however, the decrease in HDL was of significantly greater magnitude than seen in previous studies. Apo A-I autoantibodies present an additional mechanism for causing low HDL levels.Until a better understanding of the origins of the acquired HDL deficiency is attained, these results suggest that an unexplained marked decrease in HDL may indicate a developing gammopathy. Additional studies are needed to understand and define the prevalence of antibodies recognizing apo A-I, the relationship to HDL-C levels, and importantly, to examine whether such a mechanism of HDL deficiency represents a potential for cardiovascular disease risk. References 1 Kilgore LL, Patterson BW, Parenti DM, Fisher WR. Immune complex hyperlipidemia induced by an apolipoprotein-reactive immunoglobulin A paraprotein from a patient with multiple myeloma: characterization of this immunoglobulin. J Clin Invest. 1985; 76: 225–232.CrossrefMedlineGoogle Scholar2 Nozaki S, Ito Y, Nakagawa T, Yamashita S, Sasaki J, Matsuzawa Y. Autoimmune hyperlipidemia with inhibitory monoclonal antibodies against low density lipoprotein binding to fibroblasts in a case with multiple myeloma. Intern Med. 1997; 36: 920–925.CrossrefMedlineGoogle Scholar3 Groszek E, Abrams JJ, Grundy SM. Normolipidemic planar xanthomatosis associated with benign monoclonal gammopathy. Metabolism. 1981;30927–30935.Google Scholar4 Zadak Z, Tichy M. Hyperlipidemia and monoclonal gammopathy. Neoplasma. 1987; 34: 169–172.MedlineGoogle Scholar5 Lazarevic MB, Vitic J, Myones BL, Mladenovic V, Nanusevic N, Skosey JL, Swedler WI. Anti-lipoprotein antibodies in rheumatoid arthritis. Semin Arthritis Rheum. 1993; 22: 385–391.CrossrefMedlineGoogle Scholar6 Lahita RG, Rivkin E, Cavanagh I, Romano P. Low levels of total cholesterol, high-density lipoprotein, and apolipoprotein A1 in association with anticardiolipin antibodies in patients with systemic lupus erythematosus. Arthritis Rheum. 1993; 36: 1566–1574.CrossrefMedlineGoogle Scholar7 Dinu AR, Merrill JT, Shen C, Antonov IV, Myones BL, Lahita RG. Frequency of antibodies to the cholesterol transport protein apolipoprotein A1 in patients with SLE. Lupus. 1998; 7: 355–360.CrossrefMedlineGoogle Scholar8 Miida T, Kawano M, Fielding CJ, Fielding PE. Regulation of the concentration of pre beta high-density lipoprotein in normal plasma by cell membranes and lecithin-cholesterol acyltransferase activity. Biochemistry. 1992; 31: 11112–11117.CrossrefMedlineGoogle Scholar9 Levy Y, Aviram M, Spira G, Tatarsky I, Brook GJ, Carter A. Plasma cholesterol concentration and extra lipid band in monoclonal gammopathies. 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Goldberg R and Rader D (2015) The Disappearing High Density Lipoprotein Syndrome, Journal of Clinical Lipidology, 10.1016/j.jacl.2014.12.011, 9:1, (91-92), Online publication date: 1-Jan-2015. Jorge I, Burillo E, Mesa R, Baila-Rueda L, Moreno M, Trevisan-Herraz M, Silla-Castro J, Camafeita E, Ortega-Muñoz M, Bonzon-Kulichenko E, Calvo I, Cenarro A, Civeira F and Vázquez J (2014) The human HDL proteome displays high inter-individual variability and is altered dynamically in response to angioplasty-induced atheroma plaque rupture, Journal of Proteomics, 10.1016/j.jprot.2014.04.010, 106, (61-73), Online publication date: 1-Jun-2014. Ohba K, Oki Y, Fujita K, Kameko F, Moriyama T, Horiike Y, Morita H, Matsushita A, Iino K, Sasaki S, Nakamura H and Maekawa M (2013) A sudden onset and the spontaneous remission of severe hypo-high-density lipoprotein cholesterolemia without serious underlying disease: A case report, Clinica Chimica Acta, 10.1016/j.cca.2013.09.003, 426, (91-94), Online publication date: 1-Nov-2013. Stone N (2009) Clinical Evaluation for Genetic and Secondary Causes of Dyslipidemia Clinical Lipidology, 10.1016/B978-141605469-6.50016-0, (144-157), . Goldberg R and Mendez A (2007) Severe acquired (secondary) high-density lipoprotein deficiency, Journal of Clinical Lipidology, 10.1016/j.jacl.2007.02.003, 1:1, (41-56), Online publication date: 1-Mar-2007. (2007) Case 40-2006: Anemia and Low HDL Cholesterol, New England Journal of Medicine, 10.1056/NEJMc070139, 356:18, (1893-1895), Online publication date: 3-May-2007. October 2002Vol 22, Issue 10 Advertisement Article InformationMetrics https://doi.org/10.1161/01.ATV.0000032153.19645.5C Originally publishedOctober 1, 2002 PDF download Advertisement

OBJECTIVES:To characterize antineutrophil cytoplasmic antibodies (ANCA), their major autoantigens, disease associations, and pathophysiology in systemic vasculitides. To describe a patient with a novel de novo ANCA-associated vasculitis after kidney transplantation.METHODS:We reviewed and compiled the literature on ANCA-related topics and systemic vasculitis. Laboratory and clinical data from a cadaveric kidney transplant patient who developed necrotizing vasculitis involving glomerular capillaries, with crescent formation associated with P-ANCA and myeloperoxidase, were analyzed.RESULTS:Large-scale multi-center testing of patient and normal sera by the European ANCA Assay Standardization Project using immunofluorescence assays and enzyme immunoassays indicate the assays have good sensitivity and specificity, and diagnostic utility for ANCA-associated vasculitis. A few investigations covering basic and clinical research with ANCA remain controversial: whether endothelial cells do or do not express a 29-kd neutral serine protease termed proteinase-3 (PR-3), the target of ANCA in most individuals with Wegener's granulomatosis, and whether anti-myeloperoxidase (MPO) ANCAs recognize a restricted number of epitopes on MPO. This issue has relevance for using monoclonal antibodies to treat patients with vasculitis who have adverse effects from immunosuppressive drugs. The two allelic forms of FcgammaRIIa (H131/R131) and the two of FcgammaRIIlb (NA1/NA2) are discussed as possible inheritable genetic elements for vasculitic disorders and for signaling responses. Stimulatory and costimulatory molecules, and cytokine profiles of T lymphocytes are characterized to show that these cells are actively involved in the ANCA-associated vasculitides. The patient described had a de novo ANCA associated small vessel vasculitis which developed after renal transplantation.CONCLUSIONS:There have been significant advances in the development of sensitive and specific ANCA assays. The immunopathogenetic mechanism of ANCA involves the constitutive FcgammaRs, ligands, and signaling responses to activate cytokine-primed neutrophils. This may lead to the generation of reactive oxygen intermediates, degranulation, and secretion of intracellular granule contents, and ultimately inflammation and vasculitis.

The State of Florida's practice of releasing unmarked early juvenile red drum Sciaenops ocellatus L. to create a recreational fishery in Biscayne Bay, FL, USA, was assessed. Cohorts were reared in ponds to an early juvenile size of approximate to 50 mm total length. Fish were then harvested and transported immediately to release locations within 24 h. Substantial mortality often resulted after harvesting and immediate transport of early juveniles. Weak correlations between post-transport mortality, transport conditions and fish characteristics (i.e. age, size and condition) suggested that stressors operating before or during pond harvest were predetermining toleration of transport. Seine sampling 1-6 days after cohorts were released indicated that fish 'disappeared' from release sites faster than the rates of mortality observed for their siblings monitored in food-rich, predator-free tanks. Visual and immunological analyses of gut contents suggested that juvenile great barracuda Sphyraena barracuda (Walbaum) and adult redfin needlefish Strongylura notata (Poey) were the major predators. Strategies for reducing pre- and post-release mortality and directions for future research are suggested. Discontinuing the release of unmarked organisms of any type is strongly recommended.

Since our eventual goal is to distinguish with immunologic tools the early life history forms of specific species of the Lutjanidae, different fluorescent dye-antibody conjugates were investigated as a model for that utility. Immunofluorescent tests were carried out with two dyes that are excited in the near-infrared region of the spectrum, La Jolla Blue(TM) (LJB) and Cy 5.5(TM), and with two dyes excited at the lower part of the visible range, FluorX(TM) and fluorescein isothiocyanate (FITC). The LJB is a relatively new fluorescent probe that was produced from the phthalocyanine class of dyes. Each dye was covalently labeled to the IgG fraction of a goat antiserum produced to a highly purified 66 kDa glycoprotein found in soluble extracts of adults, juveniles, and oocytes of the gray snapper Lutjanus griseus. An epi-fluorescence microscope was used with a camera, image processor, color video monitor and printer, and a PC computer with software for image acquisition and light intensity measurement. First, we investigated the autofluorescence properties of the oocytes at 490 nm (low visible range) and 685 nm (near infrared). Second, because photobleaching is a phenomenon inherent in fluorescence probes, each immune IgG-fluorescent dye conjugate was incubated with the lutjanid oocytes for temporal assays at excitation wavelengths. Significant photobleaching was observed the first few minutes of excitation at 490 nm for oocytes reacted with the FluorX and FITC conjugates and for the Cy 5.5 conjugate at 685 nm. Oocytes were allowed to recover for 30 min, then re-excited at either 490 or 685 nm. The photobleaching effect was reversible only to a small degree. Little or no photobleaching occurred with La Jolla Blue during the 8 min excitation period at 685 nm, showing that the dye is superior when prolonged periods of specimen observation are necessary using fluorescence microscopy.

The presence of anti-CD36 antibodies in plasma of patients with thrombotic thrombocytopenic purpura (TTP), idiopathic thrombocytopenic purpura (ITP), and heparin-induced thrombocytopenia without/with thrombosis (HIT/HITT) has been examined by immunoblots, and a monoclonal antibody capture assay, the platelet-associated IgG characterization assay (PAICA). Results with PAICA showed that 73% (8/11) of patients with TTP were positive, and 71% (10/14) by immunoblots. With ITP, 20% (6/30) were positive by PAICA and 19% (3/16) by immunoblots; HIT, 30% (3/10) were positive by PAICA and 60% (6/10) by immunoblot; HITT. 50% (2/4) by PAICA and 100% (4/4) by immunoblot. Purification of CD36 by fast protein liquid chromatography (FPLC) from Triton X-100 extracts of normal platelet membranes resulted in the isolation of two different forms: the classic 88 kD form, and a second, lighter 85 kD form. Our data indicated that the patients' plasma autoantibodies reacted strongly with the 85 kD form. Conventional monoclonal and polyclonal antisera produced to the 88 kD form reacted strongly with the 88 kD form but weakly with the 85 kD form. These results confirm the possible importance of anti-CD36 antibodies in the pathophysiology of TTP and other thrombocytopenias and demonstrate the presence of a previously unrecognized target antigen for these antibodies.