Proximal Tubular Expression Patterns of Megalin and Cubilin in Proteinuric Nephropathies.

INTRODUCTION: Receptor-mediated endocytosis is responsible for protein reabsorption in the proximal tubules. For albumin this process involves at least 2 interacting receptors, megalin and cubilin. Albumin is not usually present in the urine, indicating a highly efficient tubular reuptake under physiological conditions. However, early appearance of albuminuria may mean that the tubular system is overwhelmed by large quantities of albumin or that the function is impaired. METHODS: To better understand the physiological role of megalin and cubilin in human renal disease, renal biopsies from 15 patients with a range of albuminuria and 3 healthy living donors were analyzed for proximal tubular expression of megalin and cubilin using immunohistochemistry (IHC) and semiquantitative immune-electron microscopy. Their expression in proteinuric zebrafish was also studied. RESULTS: Megalin and cubilin were expressed in brush border and cytoplasmic vesicles. Patients with microalbuminuric IgA nephropathy and thin membrane disease had significantly higher megalin in proximal tubules, whereas those with macro- or nephrotic-range albuminuria had unchanged levels. Cubilin expression was significantly higher in all patients. In a proteinuric zebrafish nphs2 knockdown model, we found a dose-dependent increase in the expression of tubular megalin and cubilin in response to tubular protein uptake. DISCUSSION: Megalin and cubilin show different expression patterns in different human diseases, which indicates that the 2 tubular proteins differently cooperate in cleaning up plasma proteins in kidney tubules.

Biomarkers of Cardiovascular Disease and Mortality Risk in Patients with Advanced CKD.

BACKGROUND AND OBJECTIVES: The high risk of cardiovascular disease (CVD) and premature death in patients with CKD associates with a plethora of elevated circulating biomarkers that may reflect distinct signaling pathways or simply, are epiphenomena of CKD. We compared the predictive strength of 12 biomarkers analyzed concomitantly in patients with stage 5 CKD. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: From 1994 to 2014, 543 patients with stage 5 CKD (median age =56 years old; 63% men; 199 patients had CVD) took part in our study on malnutrition, inflammation, and CVD in incident dialysis patients. Circulating levels of albumin, ferritin, high-sensitivity C-reactive protein (hsCRP), IGF-1, IL-6, orosomucoid, troponin T (TnT), TNF, soluble intracellular adhesion molecule, soluble vascular cellular adhesion molecule 1 (sVCAM-1), and platelet and white blood cell (WBC) counts were analyzed as predictors of the presence of clinically overt CVD at baseline, protein-energy wasting (PEW), and subsequent all-cause mortality. During follow-up for a median of 28 months, there were 149 deaths, 81 of which were caused by CVD. RESULTS: Most biomarkers were elevated compared with reference values and--except for albumin, ferritin, and IGF-1-higher in patients with CVD. In receiver operating characteristic analysis, age, IL-6, TnT, hsCRP, and IGF-1 were classifiers of baseline CVD and predictors of all-cause mortality. In addition to age, diabetes mellitus, smoking (for CVD), and PEW, only IL-6, relative risk (RR) 1.10 and 95% confidence interval ([95% CI], 1.02 to 1.19), sVCAM-1 RR 1.09 (95% CI, 1.01 to 1.17), and serum albumin RR 0.89 (95% CI, 0.83 to 0.95) associated with baseline CVD, and only WBC, hazard ratio (HR) 1.94 (95% CI, 1.34 to 2.82), IL-6 HR 1.79 (95% CI, 1.20 to 2.67), and TNF HR 0.65 (95% CI, 0.44 to 0.97) predicted all-cause mortality. CONCLUSIONS: In addition to age and comorbidities, only IL-6, sVCAM-1, and albumin could-independently of other biomarkers-classify clinical CVD, and only IL-6, WBC, and TNF could-independently of other biomarkers-predict all-cause mortality risk. These data underscore the robustness of IL-6 as a classifier of clinically overt CVD and predictor of all-cause mortality in patients with stage 5 CKD.

Detection of anti-IgA antibodies using the particle gel immunoassay: a rapid test for increased patient safety.

BACKGROUND: Patient safety is a major issue in transfusion medicine and commands continuous efforts to develop valid control methods aiming to avoid serious transfusion-related complications. Anti-IgA antibodies can cause anaphylactic transfusion reactions in IgA-deficient individuals. Since standard quantitative methods for anti-IgA measurement require considerable time to be performed, in an emergency situation it can be a challenge to prevent or to quickly interpret and manage acute transfusion reactions suspected to be a consequence of anti-IgA. The purpose of this study was to test and validate at our transfusion centre a rapid assay for the identification of patients with anti-IgA antibodies. MATERIALS AND METHODS: Forty-six samples (6 from healthy controls and 40 from IgA-deficient patients) were collected. Sera were analysed blindly by three different clinical laboratory technologists using two DiaMed particle gel immunoassays (ID-PaGIA) for IgA deficiency and for antibodies to IgA. The results were subsequently checked with the results of a fluorescence enzyme immunoassay conducted in the reference immunology laboratory. RESULTS: The ID-PaGIA had a sensitivity of 91.7% and specificity of 97.1% for the IgA deficiency test. With regards to the detection of anti-IgA antibodies, the sensitivity was 89.3% and the specificity 100%. The reproducibility of the test was 100%. DISCUSSION: The ID-PaGIA screening assays are suitable for the investigation of transfusion-related anaphylactic reactions in a routine blood bank laboratory. Although the gel card technique does not quantify the level of anti-IgA antibodies, it is readily available, providing an effective and simple method for the diagnosis of anti-IgA related anaphylaxis and guidance for the appropriate transfusion practice in an emergency.

New treatment for IgA nephropathy: enteric budesonide targeted to the ileocecal region ameliorates proteinuria.

BACKGROUND: Systemic corticosteroid treatment has been shown to exert some protection against renal deterioration in IgA nephropathy (IgAN) but is not commonly recommended for long-term use due to the well-known systemic side effects. In this study, we investigated the efficacy and safety of a new enteric formulation of the locally acting glucocorticoid budesonide (Nefecon®), designed to release the active compound in the ileocecal region. The primary objective was to evaluate the efficacy of targeted release budesonide on albuminuria. METHODS: Budesonide 8 mg/day was given to 16 patients with IgAN for 6 months, followed by a 3-month follow-up period. The efficacy was measured as change in 24-h urine albumin excretion, serum creatinine and estimated glomerular filtration rate (eGFR). RESULTS: The median relative reduction in urinary albumin excretion was 23% during the treatment period (interquartile range: -0.36 to -0.04, P = 0.04) with pretreatment values ranging from 0.3 to 6 g/24 h (median: 1.5 g/24 h). The median reduction in urine albumin peaked at 40% (interquartile range: -0.58 to -0.15) 2 months after treatment discontinuation. Serum creatinine was reduced by 6% (interquartile range: -0.12 to -0.02; P = 0.003), and eGFR [Modification of Diet in Renal Disease (MDRD)] increased ∼8% (interquartile range: 0.02-0.16, P = 0.003) during treatment. No major corticosteroid-related side effects were observed. CONCLUSIONS: In the present pilot study, enteric budesonide targeted to the ileocecal region had a significant effect on urine albumin excretion, accompanied by a minor reduction of serum creatinine and a modest increase of eGFR calculated by the MDRD equation, while eGFR calculated from Cockcroft-Gault equation and cystatin C was not changed. Enteric budesonide may represent a new treatment of IgAN warranting further investigation.

Permeability, ultrastructural changes, and distribution of novel proteins in the glomerular barrier in early puromycin aminonucleoside nephrosis.

BACKGROUND/AIMS: It is still unclear what happens in the glomerulus when proteinuria starts. Using puromycin aminonucleoside nephrosis (PAN) rats, we studied early ultrastructural and permeability changes in relation to the expression of the podocyte-associated molecules nephrin, α-actinin, dendrin, and plekhh2, the last two of which were only recently discovered in podocytes. METHODS: Using immune stainings, semiquantitative measurement was performed under the electron microscope. Permeability was assessed using isolated kidney perfusion with tracers. Possible effects of ACE inhibition were tested. RESULTS: By day 2, some patchy foot process effacement, but no proteinuria, appeared. The amount of nephrin was reduced in both diseased and normal areas. The other proteins showed few changes, which were limited to diseased areas. By day 4, foot process effacement was complete and proteinuria appeared in parallel with signs of size barrier damage. Nephrin decreased further, while dendrin and plekhh2 also decreased but α-actinin remained unchanged. ACE inhibition had no significant protective effect. CONCLUSIONS: PAN glomeruli already showed significant pathology by day 4, despite relatively mild proteinuria. This was preceded by altered nephrin expression, supporting its pivotal role in podocyte morphology. The novel proteins dendrin and plekhh2 were both reduced, suggesting roles in PAN, whereas α-actinin was unchanged.

Feather mites and internal parasites in small ground finches (Geospiza fuliginosa, Emberizidae) from the Galapagos Islands (Equador).

During a parasite survey, we collected data on the presence and distribution of feather mites, intestinal parasites, and blood parasites of small ground finches (Geospiza fuliginosa) from 4 islands in the Galapagos. We recorded 4 species of feather mites, with the most common species, Trouessartia geospiza, present on the majority (77% [308/400]) of individuals. Birds with high loads of T. geospiza came from larger islands and had higher body masses. We identified 3 species of intestinal Isospora (Isospora fragmenta, Isospora temeraria, and Isospora exigua) in fecal samples that showed a diurnal pattern of oocyst release. Among samples collected in the afternoon, infection prevalence was 61% (11/18), while only 0.5% (1/192) contained oocysts in the morning. We screened 40 individuals from one island (Isabela) for blood parasites using molecular markers. Although no parasites of Haemoproteus, Leucocytozoon, or Plasmodium were detected, a high proportion of birds (80% [32/40]) had systemic Isospora spp. infections. A high infection prevalence (74% [20/27]), but low infection intensity, was confirmed using optical microscopy. This result could either be due to the detection of a previously unidentified systemic Isospora sp. parasite, or a result of the previously described Isospora spp. parasites causing systemic infections.

Experimentally increased social competition compromises humoral immune responses in house finches.

Although social behavior can substantially influence an individual's physiology, few studies have examined whether intraspecific competition compromises individual immunocompetence. We experimentally manipulated the intensity of social competition in captive non-breeding house finches (Carpodacus mexicanus) by supplying few (high competition) or many (low competition) feeding sites. We tested whether elevated levels of social competition caused individual changes in aggression rates, humoral immunity, body mass, and baseline and stress-induced corticosterone concentrations. We also examined whether physiological responses to social competition were related to an individual's social status. We found that house finches under high social competition had significantly higher aggression rates, lower antibody responses, and lost more body mass. Within flocks, dominant individuals mounted stronger immune responses in both competition treatments. Our statistical power to detect differences in circulating corticosterone concentrations was low, but we did not find any support for the hypothesis that corticosterone concentrations mediate immunosuppression among or within flocks: baseline and stress-induced corticosterone concentrations did not differ under high and low social competition, were unrelated to individual social status, and did not predict the extent of immunosuppression among individuals. Overall, we documented that two universal components of social behavior, intraspecific competition and social status, modulated the strength of a humoral immune response in house finches.

Stress responses and disease in three wintering house finch (Carpodacus mexicanus) populations along a latitudinal gradient.

In laboratory studies, stress hormones have been shown to impair immune functions, and increase susceptibility to diseases. However, the interactions between stress hormones and disease have rarely been studied in free-ranging populations. In this study, we measured concentrations of the avian stress hormone corticosterone across four winter months (December-March) over two years in three eastern North American house finch populations (Carpodacus mexicanus) along a latitudinal gradient. Because Mycoplasma gallisepticum infections appear in these populations in late winter, we hypothesized that the timing of the disease outbreaks could be mediated by changes in corticosterone concentrations. We found a significant increase in baseline and stress-induced plasma corticosterone concentrations in house finches without Mycoplasma symptoms in late winter; when the prevalence of Mycoplasma infection peaks. We also found that house finches with Mycoplasma symptoms had elevated stress-induced corticosterone concentrations. High baseline concentrations were associated with a low body condition and a high fat load. We found that the relationship between corticosterone concentrations and the latitude of the study population changed between years. The first year, corticosterone concentrations were lowest in the southern latitude, but became higher in the second year when average winter temperatures were low. A causal understanding of the implications for this variation in corticosterone concentrations for Mycoplasma disease dynamics awaits further studies.

House sparrows (Passer domesticus) adjust their social status position to their physiological costs.

For group-living animals, the maintenance of a position in the social hierarchy may be associated with physiological costs such as increased stress and energy expenditure or suppressed immune functions. In this study, we experimentally manipulated the social status of house sparrows so that each bird experienced two social environments in random sequence: being dominant and subordinate. For 14 males, we investigated how corticosterone concentrations, energy expenditure and immune functions were affected by these changes in social status position. We found that the cost of maintaining a social status position differed between individuals and were related to individual body size. Birds with small body size had increased costs in terms of increased stress responses and reduced cell-mediated immune responses while being experimentally kept as dominants, while birds with large body size had increased costs while they were subordinates. We also found that birds with increased energetic and immunological costs as dominants obtained a low status position in the large group, while birds with increased costs as subordinates obtained a high status position in the large group. In summary, we found that the costs associated with the maintenance of social status position differed between individuals and was related to the individuals' body size. Furthermore, in a large group, individuals maintained a social status position that minimized energetic and immunological costs.

Mites associated with the small ground finch, Geospiza fuliginosa (Passeriformes: Emberizidae), from the Galápagos Islands.

In collections of ectoparasites from 368 small ground finches, Geospiza fuliginosa, in populations from the islands of Isabela, Santa Cruz, San Cristóbal, and Santa Fé, in the Galápagos Archipelago, Ecuador, we found 8 species of mites. Four mite species were common on all islands sampled, i.e., Mesalgoides geospizae Mironov and Pérez (Psoroptoididae), Xolalges palmai Mironov and Pérez (Xolalgidae), and 2 new species, Trouessartia geospiza n. sp. (Trouessartiidae) and Proctophyllodes darwini n. sp. (Proctophyllodidae). Four other species were represented by single collections from G. fuliginosa, i.e., Pterodectes atyeoi n. sp. (Proctophyllodidae), Strelkoviacarus sp. (Analgidae), Dermoglyphus sp. (Dermoglyphidae), and Dermanyssus sp. (Dermanyssidae). Authorship of new species names is attributed to the 3 authors who prepared the descriptions (B.M.O.C., J.F., D.L.). Trouessartia geospiza and P. atyeoi were also found on previously collected specimens of other Geospiza species in museum collections. For the 4 common species, we found no differences in prevalence among the 4 island populations, but infection prevalence differed among the 4 species. The mean infection prevalence was high for T. geospizae (89%), moderate for M. geospizae (58%) and X. palmai (44%), and low for P. darwini (26%) in all populations. The feather mite fauna of G. fuliginosa was similar to that of other Geospiza species, and generally related to communities found on other emberizid finches.

Binding of adenosine receptor ligands to brain of adenosine receptor knock-out mice: evidence that CGS 21680 binds to A1 receptors in hippocampus.

The adenosine receptor agonist 2-[ p-(2-carboxyethyl)phenylethylamino]-5'- N-ethylcarboxamidoadenosine (CGS 21680) is generally considered to be a selective adenosine A(2A) receptor ligand. However, the compound has previously been shown to exhibit binding characteristics that are not compatible with adenosine A(2A) receptor binding, at least in brain regions other than the striatum. We have examined binding of [(3)H]CGS 21680 and of antagonist radioligands with high selectivity for adenosine A(1) or A(2A) receptors to hippocampus and striatum of mice lacking either adenosine A(1) (A1R((-/-))) or A(2A) (A2AR((-/-))) receptors. Both receptor autoradiography and membrane binding techniques were used for this purpose and gave similar results. There were no significant changes in the binding of the A(1) receptor antagonist [(3)H]DPCPX in mice lacking A(2A) receptors, or in the binding of the A(2A) receptor antagonists [(3)H]SCH 58261 and [(3)H]ZM 241385 in mice lacking A(1) receptors. Furthermore, [(3)H]CGS 21680 binding in striatum was abolished in the A2AR((-/-)), and essentially unaffected in striatum from mice lacking A(1) receptors. In hippocampus, however, binding of [(3)H]CGS 21680 remained in the A2AR((-/-)), whereas binding was virtually abolished in the A1R((-/-)). There were no adaptive alterations in A(2A) receptor expression in this region in A1R((-/-)) mice. Thus, most of the [(3)H]CGS 21680 binding in hippocampus is dependent on the presence of adenosine A(1) receptors, but not on A(2A) receptors, indicating a novel binding site or novel binding mode.

Immunological investments reflect parasite abundance in island populations of Darwin's finches.

The evolution of parasite resistance can be influenced by the abundance of parasites in the environment. However, it is yet unresolved whether vertebrates change their investment in immune function in response to variation in parasite abundance. Here, we compare parasite abundance in four populations of small ground finches (Geospiza fuliginosa) in the Galapagos archipelago. We predicted that populations exposed to high parasite loads should invest more in immune defence, or alternatively use a different immunological defence strategy. We found that parasite prevalence and/or infection intensity increased with island size. As predicted, birds on large islands had increased concentrations of natural antibodies and mounted a strong specific antibody response faster than birds on smaller islands. By contrast, the magnitude of cell-mediated immune responses decreased with increasing parasite pressure, i.e. on larger islands. The data support the hypothesis that investments into the immune defence are influenced by parasite-mediated selection. Our results are consistent with the hypothesis that different immunological defence strategies are optimal in parasite-rich and parasite-poor environments.