Caratterizzazione di EPC in pazienti con malattia renale cronica. Background: Gli effetti del recettore della vitamina D (VDR)e dell'espressione dell' osteocalcina (OCN), così come VDR agonista terapia (VDRA) su circolanti cellule progenitrici endoteliali (EPCs) non è stato ancora chiarito. Metodi: sono state analizzat EPCs in 23 controlli sani e 53 pazienti sottoposti a dialisi. La percentuale di EPCs (CD34+KDR+CD133+/-CD45-) VDR+/- o OCN+/- è stata analizzata in citometria a flusso e correlata con molecole coinvolte nelle malattie. Risultati: EPCs aumentano nei pazienti con CKD trattati con VDRAs. EPCsOCN +/- in pazienti non trattati con VDRAs correlano positivamente con il calcio sierico e reticolociti, e negativamente con DKK1. La percentuale di EPCsVDR +/- correla negativamente con OPN.  EPCsOCN + nei pazienti trattati con VDRAs correla positivamente con la vitamina D. La percentuale di EPCsVDR +/- positivamente correla con IL-6. Conclusioni: la terapia VDRA influenza l'espressione VDR e di OCN su EPCs circolanti. Dal momento che l'espressione OCN può contribuire alla calcificazione vascolare, ipotizziamo un putativo effetto pro-calcificazione di VDRA. Effetti immunomodulatori di EPO. Background: Il cloruro di sodio spinge l'induzione di cellule TH17 patogeni e TH1. La correzione dell'anemia con eritropoietina (EPO) è associato ad un miglioramento della tolleranza del trapianto di rene. Prove emergenti indicano che queste osservazioni possano essere eritropoiesi-indipendente e che l'EPO presenta proprietà immunosoppressive. Metodi: esaminato gli effetti del trattamento con EPO e sale su cellule T umana, apoptosi e la produzione di INF-γ, l'effetto sulla iTh17 e iFoxP3. Le cellule sono state analizzate con l'analisi di citometria a flusso. Risultati: NaCl aumenta la proliferazione di CD4+ e CD8+, iTh17 e la produzione di INFγ. Questo effetto è prevenuto da EPO. EPO aumenta l'induzione di Foxp3. Non cambia l'apoptosi e la stabilità di FoxP3. EPO è in grado di contiene l'effetto pro infiammatorio del sale / First project: Characterization of EPCs in patients with CKD. Background: The effects of vitamin D receptor (VDR) and osteocalcin (OCN) expression as well as VDR agonist (VDRA) therapy on circulating endothelial progenitor cells (EPCs) has not been elucidated yet. Methods: we therefore analyzed EPCs in 23 healthy controls and 53 patients undergoing dialysis. The percentage of EPCs (CD34+CD133+/-KDR+CD45-) expressing VDR or OCN were analyzed using flow cytometry and correlated with molecules involved in diseases. Results: EPCs increase in CKD patients treated with VDRAs. EPCsOCN+/- in patients untreated with VDRAs correlated positively with serum calcium and reticulocytes, and negatively with DKK1. The percentage of EPCsVDR+/- correlated negatively with OPN. EPCsOCN+ in patients treated with VDRAs correlated positively with vitamin D. The percentage of EPCsVDR+/- correlated positively with IL-6. Conclusions: our data suggest that VDRA therapy influence VDR and OCN expression on circulating EPCs. Since OC expression may contribute to vascular calcification, we hypothesize a putative pro-calcifying effect of VDRA. Second project: Immunomodulatory effects of EPO. Background: Sodium chloride drives the induction of pathogenic TH17 cells and TH1. Correction of anemia with erythropoietin (EPO) is associated with improved kidney transplant outcomes. Emerging evidence indicates that these observations may be erythropoiesis-independent and that EPO exhibits immunosuppressive properties. Methods: examined the effects of treatment with EPO and salt on human T-cell alloimmunity, the apoptosis and the production to INF-γ, the effect on iTh17 and iFoxP3. The cells were analyzed with flow cytometry analysis. Results: NaCl increases the proliferation of CD4+ and CD8+ cells, iTh17 and the production of INFγ. This effect is prevented by EPO. EPO increases the induction of Foxp3. It does not change the apoptosis and stability of FoxP3. Epo contains the inflammatory effect of NaCl. EPO in the kidney, where NaCl is high, may have a tolerance effect.

Made available in DSpace on 2019-03-30T00:31:12Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-12-07 / Introduction: Glomerulopathies are diseases that mainly affect young adults between 20 and 40 years. Recently, serum levels of syndecan-1, a biomarker of endothelial glycocalyx damage, have been shown to be increased in nephrotic patients with mild loss of renal function and is important for endothelial dysfunction in these patients. In this study, we assessed the level of angiopoietin-2 syndrome (PTGA2) in patients with nephrotic syndrome and near normal renal function and its possible interaction with endothelial glycocalyx derangement. Objective: To evaluate the new biomarkers and endotelial lesion in patients with nephrotic syndrome and the role of nephrotic endothelial-angiopioetin-2 syndrome. Method: A cross-sectional study conducted from January to November 2017. Adult patients (age> 18 years) with nephrotic syndrome and without immunosuppression were included. Samples of blood after 12 h fasting for further measurement of syndecan-1 and AGPT2. Mediation analyzes were performed to evaluate the hypothetical associations of nephrotic syndrome and AGPT2 syndrome with syndecan-1. Results: 65 patients were included, of which 37 (56.9%) were females with primary glomerular disease. The levels of Syndecan-1 in nephrotic patients were higher than in the control group (102.8 ± 36.2 vs. 28.2 ± 9.8 ng / mL, p <0.001). Adjusted syndecan-1 correlation with features of major nephrotic syndrome after adjustments for age and eGFR are demonstrated. Syndecan-1 was significantly associated with 24-hour urinary protein excretion, total cholesterol, LDL-cholesterol, HDL-cholesterol, and triglycerides. Angiopoietin-2 was independently associated with serum albumin, excretion of 24-hour urinary proteins, total cholesterol and LDL-cholesterol, and a strong association with syndecan-1 (0.461, p <0.001). The results of the mediation analyzes showed a direct association between LDL-cholesterol and syndecan-1 remained non-significant after the inclusion of AGPT-2 in the mediation analysis and AGPT2 explained 56% of the total association observed between LDL-cholesterol and syndecan-1. Conclusion: The association between LDL-cholesterol and glycocalyx derangement in nephrotic patients is mediated by AGPT2. Keywords: Nephrotic Syndrome; Primary Glomerulopathies; Glycopaloid; Biomarkers. / Introdução: As glomerulopatias são doenças que afetam principalmente adultos jovens entre 20 e 40 anos. Recentemente, demonstrou-se que os níveis séricos de syndecan-1, um biomarcador do dano do glicocálix endotelial, estão aumentados em pacientes nefróticos com perda discreta da função renal e é importante para a disfunção endotelial nesses pacientes. Neste trabalho, avaliamos o nível de síndrome de angiopoietina-2 (AGPT2) em pacientes com síndrome nefrótica e função renal quase normal e sua possível interação com o desarranjo do glicocálix endotelial. Objetivo: Avaliar os novos biomarcadores de lesão endotelial em pacientes com síndrome nefrótica e o papel da síndrome nefrótica angiopioetina-2 endotelial. Método: Foi realizado um estudo transversal realizado de janeiro a novembro de 2017. Foram incluídos pacientes adultos (idade > 18 anos) com síndrome nefrótica e sem imunossupressão. As amostras de sangue após jejum de 12 h para posterior mensuração de syndecan-1 e AGPT2. Análises de mediação foram realizadas para avaliar as associações hipotéticas de características da síndrome nefrótica e AGPT2 com syndecan-1. Resultados: Foram incluídos 65 pacientes, sendo 37 (56,9%) do sexo feminino com doença glomerular primária. Os níveis de syndecan-1 em pacientes nefróticos foram maiores que no grupo controle (102,8 ± 36,2 vs. 28,2 ± 9,8 ng/mL, p <0,001). Correlação ajustada de syndecan-1 com características da síndrome nefrótica principal após ajustes para idade e TFGe são demonstrados. Syndecan-1 foi significativamente associado com a excreção de proteína urinária de 24 h, colesterol total, LDL-colesterol, HDL-colesterol e triglicérides. A angiopoietina-2 foi independentemente associada à albumina sérica, excreção de proteínas urinárias de 24 horas, colesterol total e LDL-colesterol, além de uma forte associação com syndecan-1 (0,461, p < 0,001). Os resultados das análises de mediação mostram associação direta entre LDL-colesterol e syndecan-1 permaneceu não significativa após a inclusão de AGPT-2 na análise de mediação e AGPT2 explicou 56% da associação total observada entre LDL-colesterol e syndecan-1. Conclusão: A associação entre LDL-colesterol e desarranjo do glicocálice em pacientes nefróticos é mediada pelo AGPT2. Palavras-Chave: Síndrome nefrótica; Glomerulopatias primária; Glicocálix; Biomarcadores.

O trabalho que a seguir se apresenta tem como principal enfoque a terminologia dos termos utilizados pelos profissionais de nefrologia. A abordagem e os métodos de pesquisa foram aqueles da socioterminologia, que consagra as diferentes formas de utilização de termos que os diversos falantes de uma língua empregam, especialmente no contexto de um domíniio específico Estas alterações podem ser resultantes do grupo etário, social ou da região geográfica dos seus utilizadores. Para tal realizou-se um questionário que foi respondido por 43 pessoas de vários grupos etários, profissionais e de diferentes zonas geográficas. Neste trabalho, pretendeu-se essencialmente detectar situações menos correctas do ponto de vista terminológico e apontar algumas soluções para a harmonização terminológica da nefrologia em português.

Simkania negevensis is a bacterium belonging to the order Chlamydiales but with certain biological characteristics different from those of chlamydia, according to which it was classified in the family Simkaniaceae. It is widespread in the environment, due to its ability to survive in amoebae also in phase cystic, for which it was hypothesized a possible transmission after contact with water in which they are present amoebae. So far it is known its role in diseases of the lower respiratory tract, such as childhood bronchiolitis and pneumonia in adults of the community, following its transmission through infected aerosols. A recent American study showed, by PCR, a high prevalence of S. negevensis in patients with lung transplant than other transplant recipients, assuming an association between the presence of the bacterium in these patients, and transplant rejection, were more frequent in lung transplant recipients infected compared to uninfected. There are no data so far analyzed in Italy relative to the population of dialysis and kidney transplant recipients relative to simkania negevensis why this study was undertaken in order to start a specific location and evaluate the scientific implications. Because its ability to assume persistent forms of infection, which may lead to a prolonged inflammatory response, Simkania negevensis, similar to other persistent bacteria or viruses, may be ivolved in pathologic complication. Sn may be a factor in graft rejection in mmunesuppressed lung transplant recipients, and further studies are planned to explore the posible association of Sn infections with various in vivo pathologies.

Introduction. Cardiovascular disease (CVD) represents the main cause of morbidity and mortality in kidney recipients. This study was undertaken to assess the impact of functional polymorphisms located in cytokine and apoptosis genes on CVD after kidney transplantation. Cytokine polymorphisms, generally located in gene regulatory regions, are associated with high and low cytokine production and are likely to modulate the magnitude of inflammatory responses following transplantation, depending on the balance between the levels of pro-inflammatory and antiinflammatory cytokines. The role of apoptosis in atherosclerosis has not been completely elucidated, and here we explored the hypothesis that the heterogeneity in cardiovascular risk in kidney recipients may also be linked to functional polymorphisms involved in apoptosis induction. Purpose. In the search for relevant genetic markers of predisposition to CVD after renal transplant, the present investigation was undertaken to identify the clinical impact of polymorphisms of cytokines TNF-α, TGF-β, IL-10, IL-6, IFN-γ and IL-8 and of apoptosis genes Fas and Caspase 9 in a population of kidney transplant recipients. Materials and methods. The study involved 167 patients who received cadaveric kidney transplantation at our centre between 1997 and 2005 (minimum follow-up of 12 months); 35 of them had experienced cardiovascular events (CVD group) and 132 had no cardiovascular complications (non-CVD group). Genotyping was performed using RFLP (Restriction Fragment Length Polymorphism) for RFLP per IL-8/T-251A, Fas/G-670A e Casp9/R221Q polymorphism and SSP (Sequence Specific Primer) for TNF-α/G-308A, TGF-β/L10P, TGF-β/R25P, IL-10/G-1082A, IL- 10/C-819T, IL-10/C-592A, IL-6/G-174C, IFN-γ/T+874A polymorphisms.Results. We found a significant difference in TNF-α and IL-10 genotype frequencies between the patients who had suffered cardiovascular events and those with no CVD history. The high producer genotype for proflogistic cytokine TNF-α appeared to have a significantly superior prevalence in the CVD group compared to the non-CVD group (40.0% vs 21.2%) and it resulted in a 2.4-fold increased cardiovascular risk (OR=2.361; p=0.0289). On the other hand, the high producer genotype for the antiinflammatory cytokine IL-10 was found in 2.8% of the CVD group and in 16.7% of non-CVD group; logistic regression showed a 0.3-fold reduced risk of CVD associated with genetically determined high IL-10 production (OR=0.278; p<0.0001). The other polymorphisms did not prove to have any impact on CVD. Conclusions. TNF-α and IL-10 gene polymorphisms might represent cardiovascular risk markers in renal transplant recipients.

Uric acid is a major inducer of inflammation in renal interstitium and may play a role in the progression of renal damage in hyperuricemic subjects with primary nephropathies, renal vascular disease, and essential hypertension. At the same time, UA also acts as a water-soluble scavenger of reactive oxygen species. We evaluated the cellular effects of UA on cultured HMC as a potential interstitial target for abnormally elevated levels in acute and chronic renal disease. Intracellular free Ca2+ ([Ca2+]i) was monitored by microfluorometry of fura 2-loaded cells, while oxidation of intracellularly trapped non-fluorescent 2’,7’-dichlorofluorescein diacetate (DCFHDA, 20 uM) was employed to assess the generation of reactive oxygen species during 12-hr incubations with various concentrations of UA or monosodium urate. Fluorescent metabolites of DCFH-DA in the culture media of HMC were detected at 485/530 nm excitation/emission wavelengths, respectively. UA dose-dependently lowered resting [Ca2+]i (from 102±9 nM to 95±3, 57±2, 48±6 nM at 1-100 uM UA, respectively, p <0.05), leaving responses to vasoconstrictors such as angiotensin II unaffected. The effect was not due to Ca2+/H+ exchange upon acidification of the bathing media, as acetate, glutamate, lactate and other organic acids rather increased [Ca2+]i (to max. levels of 497±42 nM with 0.1 mM acetate). The decrease of [Ca2+]i was abolished by raising extracellular Ca2+ and not due to effects on Ca2+ channels or activation of Ca2+-ATPases, since unaffected by thapsigargin. The process rather appeared sensitive to removal of extracellular Na+ in combination with blockers of Na+/Ca2+ exchange, such as 2’,4’-dichlorobenzamil, pointing to a countertransport mechanism. UA dose-dependently prompted the extracellular release of oxidised DCFH (control 37±2 relative fluorescence units (RFU)/ml, 0.1uM 47±2, 1 uM 48±2, 10 uM 51±4, 0.1 mM 53±4; positive control, 10 uM sodium nitroprusside 92±5 RFU/ml, p<0.01). In summary, UA interferes with Ca2+ transport in cultured HMC, triggering oxidative stress which may initiate a sequence of events leading to interstitial injury and possibly amplifying renal vascular damage and/or the progression of chronic disease.

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Background. Mesenchymal stem cells (MSC) may be of value in regeneration of renal tissue after damage, however lack of biological knowledge and variability of results in animal models limit their utilization. Methods. We studied the effects of MSC on podocytes ‘in vitro’ and ‘in vivo’ utilizing adriamycin (ADR) as a model of renal toxicity. The ‘in vivo’ experimental approach was carried out in male Sprague Dawley rats (overall 60 animals) treated with different ADR schemes to induce acute and chronic nephrosis. MSC were given a) concomitantly to ADR in tail vein or b) in aorta and c) in tail vein 60 days after ADR. Homing was assessed with PKH26-MSC. Results. MSC rescued podocytes from apoptosis induced by ADR ‘in vitro’. The maximal effect (80% rescue) was obtained with MSC/Podocytes co-culture ratio of 1:1 for 72 hours. All rats treated with ADR developed nephrosis. In no case MSC modified the clinical parameters (i.e. proteinuria, serum creatinine, lipids) but protected the kidney from severe glomerulosclerosis when given concomitantly to ADR. Rats given MSC 60 days after ADR developed the same severe renal damage. Only few MSC were found in renal tubule-interstitial areas after 1-24 hours from injection and no MSC was detected in glomeruli. Conclusions. MSC reduced apoptosis of podocytes treated with ADR ‘in vitro’. Early and repeated MSC infusion blunted glomerular damage in chronic ADR nephropathy. MSC did not modify proteinuria and progression to renal failure, that implies lack of regenerative potential in this model.