Danilo Lofaro

Impact of body mass index on graft loss in normal and overweight patients: retrospective analysis of 206 renal transplants

Papalia T, Greco R, Lofaro D, Maestripieri S, Mancuso D, Bonofiglio R. Impact of body mass index on graft loss in normal and overweight patients: retrospective analysis of 206 renal transplants. 
Clin Transplant 2010: 24: E241–E246.

High doses of hydroxytyrosol induce apoptosis in papillary and follicular thyroid cancer cells

PurposeRecent evidences indicates that hydroxytyrosol, one of the main olive oil phenols, possess antitumor effects because of its pro-oxidant properties and the capacity to inhibit proliferation and to promote apoptosis in several tumor cell lines, although most of the results were obtained for breast and digestive systems cancers.MethodsIn this study, we evaluated the activities of hydroxytyrosol against papillary (TPC-1, FB-2) and follicular (WRO) thyroid cancer cell lines.ResultsCellular viability revealed that high doses of hydroxytyrosol reduced cancer cells viability concomitantly with a reduction of cyclin D1 expression and an up-regulation of cell cycle key modulator p21 levels. In the same experimental conditions, Annexin V–PI staining and DNA laddering revealed that hydroxytyrosol exerts proapoptotic effects on papillary and follicular cancer cells. Furthermore, by Western blot analysis, we observed that hydroxytyrosol treatment reduced thyroid cancer cells viability by promoting apoptotic cell death via intrinsic pathway.ConclusionsCollectively, our results demonstrated for the first time that in thyroid cancer cells hydroxytyrosol promoted apoptosis at higher doses with respect to other cancer cells lines. Therefore, further studies will reveal the mechanisms by which thyroid cancer cells are more resistant to the proapoptotic effect exerted by hydroxytyrosol as well as the potential application as novel target therapeutic in thyroid cancer.

Exposure to Nerve Growth Factor Worsens Nephrotoxic Effect Induced by Cyclosporine A in HK-2 Cells

Nerve growth factor is a neurotrophin that promotes cell growth, differentiation, survival and death through two different receptors: TrkANTR and p75NTR. Nerve growth factor serum concentrations increase during many inflammatory and autoimmune diseases, glomerulonephritis, chronic kidney disease, end-stage renal disease and, particularly, in renal transplant. Considering that nerve growth factor exerts beneficial effects in the treatment of major central and peripheral neurodegenerative diseases, skin and corneal ulcers, we asked whether nerve growth factor could also exert a role in Cyclosporine A-induced graft nephrotoxicity. Our hypothesis was raised from basic evidence indicating that Cyclosporine A-inhibition of calcineurin-NFAT pathway increases nerve growth factor expression levels. Therefore, we investigated the involvement of nerve growth factor and its receptors in the damage exerted by Cyclosporine A in tubular renal cells, HK-2. Our results showed that in HK-2 cells combined treatment with Cyclosporine A + nerve growth factor induced a significant reduction in cell vitality concomitant with a down-regulation of Cyclin D1 and up-regulation of p21 levels respect to cells treated with Cyclosporine A alone. Moreover functional experiments showed that the co-treatment significantly up-regulated human p21promoter activity by involvement of the Sp1 transcription factor, whose nuclear content was negatively regulated by activated NFATc1. In addition we observed that the combined exposure to Cyclosporine A + nerve growth factor promoted an up-regulation of p75 NTR and its target genes, p53 and BAD leading to the activation of intrinsic apoptosis. Finally, the chemical inhibition of p75NTR down-regulated the intrinsic apoptotic signal. We describe two new mechanisms by which nerve growth factor promotes growth arrest and apoptosis in tubular renal cells exposed to Cyclosporine A.

Adiponectin secreted by tubular renal cells during LPS exposure worsens the cellular inflammatory damage.

The pathogenetic role of adiponectin (ADPN) in kidney failure is not yet elucidated, since in vitro and in vivo studies have demonstrated that ADPN exerts both anti-inflammatory and pro-inflammatory effects. Starting from our previous findings demonstrating that HK-2 cells express and secrete ADPN, in this study we investigated the autocrine role of ADPN in tubular inflammatory damage induced by lipopolysaccharide (LPS) and the underlying molecular mechanisms. Firstly, we observed that short-term exposure to LPS enhanced ADPN protein expression as well as the adiponectin receptor ADIPOR1 mRNA content together with its signaling pathway downstream, pAMPK/pERK/pJNK, whose up-regulation status was reversed when ADPN gene knockdown occurred. Interestingly, in the same experimental conditions, we observed that ADPN mediated the nuclear translocation of the transcription factors nuclear factor kappa B (NFkB) and pcFos/pcJun (activator protein 1, AP-1), both induced by the pJNK pathway and involved in tumor necrosis factor (TNF)-α transactivation. Indeed, by transient transfection assay, we observed that the LPS-induced increase of TNF-α promoter activity was abrogated in cells pretreated with the inhibitors of NFkB and AP-1. Collectively our results suggest that in HK-2 cells, ADPN produced upon LPS stimulus could worsen the inflammatory damage in an autocrine-dependent manner.

Disentangling the Impact of Chronic Kidney Disease, Anemia, and Mobility Limitation on Mortality in Older Patients Discharged From Hospital

BACKGROUNDS Chronic kidney disease (CKD), anemia, and mobility limitation are important predictors of mortality. We aimed at investigating the interactions between estimated glomerular filtration rate (eGFR), anemia, and physical performance on 1-year mortality in older patients discharged from acute care hospitals. METHODS Four hundred and eighty seven patients enrolled in a multicenter, prospective observational study were included in the analysis. eGFR was estimated by the Berlin Initiative Study 1 equation. Anemia was defined on the basis of hemoglobin values. Mobility limitation was rated by the Short Physical Performance Battery (SPPB). Covariates included demographics, nutritional status, cognitive performance, and comorbidity. The outcome of the study was mortality over 1-year follow-up. Interactions among study variables were investigated by survival tree analysis. RESULTS eGFR < 30 mL/min/1.73 m(2), anemia, and SPPB = 0-4 were significantly associated with mortality, as were hypoalbuminemia and cognitive impairment. Survival tree analysis showed that compared to patients with SPPB ≥ 4 and eGFR ≥ 46.7 mL/min/1.73 m(2) (ie, patients with the least mortality), patients with SPPB < 4 and hemoglobin < 12.2 g/dL had the highest risk of mortality [hazard ratio (HR) = 28.9, 95%CI 10.3-81.2]. Patients with SPPB ≥ 4 and eGFR < 46.7 mL/min/1.73 m(2) and those with SPPB > 4, hemoglobin ≥ 12.2g/dL, and eGFR ≥ 58.6 mL/min/1.73 m(2) had intermediate risk (HR = 6.58, 95%CI = 2.15-20.2, and HR = 15.11, 95%CI=4.42-51.7, respectively). Having SPPB < 4, hemoglobin ≥ 12.2 g/dL, and eGFR<58.6 mL/min/l.73 m(2) was not significantly associated with increased mortality (HR = 2.95, 95%CI = 0.74-11.8). CONCLUSIONS Interactions among eGFR, anemia, and mobility limitation define different profiles of risk in older patients discharged from acute care hospitals, which deserve to be considered to identify patients needing special care and careful follow-up after discharge.

Early cytomegalovirus-specific T-cell response and estimated glomerular filtration rate identify patients at high risk of infection after renal transplantation

Assessing the risk of cytomegalovirus (CMV) viremia in kidney transplant recipients (KTR) may be helpful to indicate in which patient it is worth starting antiviral treatment during preemptive strategy.

Anthropometric measures can better predict high blood pressure in adolescents.

BACKGROUND Among children, obesity and overweight may be predictors of cardiovascular (CV) risk. The purpose of this study was to examine whether body mass index (BMI), waist circumference (WC) and waist to height ratio (WHtR) were related to blood pressure (BP) among healthy southern Italian students enrolled in 3 different secondary schools. METHODS Weight, height, BP and WC were measured; BMI and WHtR were calculated for 872 Italian students. Based on percentiles of BMI, the subjects were classified as underweight, normal weight, overweight or obese. Systolic BP or diastolic BP >95th percentile were considered as high BP values (according to the 2004 guidelines of the US National Heart, Lung, and Blood Institute). Central obesity was defined as WC >75th percentile or WHtR =0.5. RESULTS Of the students, 8.7% were obese, 29% with WC >75th percentile and 29.5% with WHtR >0.5, while 4.6% showed high BP. Logistic regression showed a strong correlation between BMI and high BP (odds ratio [OR] = 1.030, p<0.0001), between WC and high BP (OR = 1.029, p<0.0001). Also WHtR (OR = 3.403, p<0.0001) was shown to be a predictor of high BP. In the male group, all of the variables considered showed a good capability to predict high BP, while in the females, only BMI (OR = 1.019, p<0.05) and WHtR (OR = 2.685, p<0.05) were associated with high BP. CONCLUSIONS In this study, we found a different correlation between BMI, WC and BP in the 2 subgroups: males and females. Only WHtR showed a significant ability to predict high BP in both groups. WHtR might represent an easily measurable anthropometric index and a better predictor of CV risk in adolescents.

Rapamycin-induced autophagy protects proximal tubular renal cells against proteinuric damage through the transcriptional activation of the nerve growth factor receptor NGFR

ABSTRACT Experimental evidence demonstrated that macroautophagy/autophagy exerts a crucial role in maintain renal cellular homeostasis and represents a protective mechanism against renal injuries. Interestingly, it has been demonstrated that in the human proximal tubular renal cell line, HK-2, the MTOR inhibitor rapamycin enhanced autophagy and mitigated the apoptosis damage induced by urinary protein overload. However, the underlying molecular mechanism has not yet been elucidated. In our study we demonstrated, for the first time, that in HK-2 cells, the exposure to low doses of rapamycin transactivated the NGFR promoter, leading to autophagic activation. Indeed, we observed that in HK-2 cells silenced for the NGFR gene, the rapamycin-induced autophagic process was prevented, as the upregulation of the proautophagic markers, BECN1, as well as LC3-II, and the autophagic vacuoles evaluated by transmission electron microscopy, were not found. Concomitantly, using a series of deletion constructs of the NGFR promoter we found that the EGR1 transcription factor was responsible for the rapamycin-mediated transactivation of the NGFR promoter. Finally, our results provided evidence that the cotreatment with rapamycin plus albumin further enhanced autophagy via NGFR activation, reducing the proapoptotic events promoted by albumin alone. This effect was prevented in HK-2 cells silenced for the NGFR gene or pretreated with the MTOR activator, MHY1485. Taken together, our results describe a novel molecular mechanism by which rapamycin-induced autophagy, mitigates the tubular renal damage caused by proteinuria, suggesting that the use of low doses of rapamycin could represent a new therapeutic strategy to counteract the tubule-interstitial injury observed in patients affected by proteinuric nephropathies, avoiding the side effects of high doses of rapamycin.

Active compounds extracted from extra virgin olive oil counteract mesothelial-to-mesenchymal transition of peritoneal mesothelium cells exposed to conventional peritoneal dialysate: in vitro and in vivo evidences

During peritoneal dialysis (PD), peritoneal mesothelial cells undergo a transition from an epithelial phenotype to a mesenchymal phenotype that, together with the inflammatory process, promotes tissue fibrosis and a failure of peritoneal membrane function. To date, there is no definitive treatment for the progressive thickening and angiogenesis of the peritoneal membrane associated with PD. In this study we tested, in vitro and in vivo, the ability of active compounds extracted from extra virgin olive oil (AC-EVOO) to counteract the mesothelial-to-mesenchymal transition process (MMT) observed in mesothelial cells chronically exposed to the conventional peritoneal dialysate (DL). In particular, we used a cultivar from southern Italy known to have a high polyphenol content. Our results showed that, in mesothelial cells exposed to DL, the combined treatment with AC-EVOO prevented the genic and protein upregulation of key mesenchymal and inflammatory markers, as well as the MCs’ migratory capacity. Concomitantly, we tested the antifibrotic efficacy of AC-EVOO in mesothelial cells obtained from effluents of patients undergoing PD, whose “fibroblast-like” phenotype was defined by flow-cytometry assay. We observed that in these cells AC-EVOO significantly mitigated, but did not reverse, the MMT process. In conclusion, our preliminary results suggest that AC-EVOO can interfere with critical factors in the process of differentiation, preventing myofibroblast formation, but once fibrosis has already progressed it is unable to promote the redifferentiation to the epithelial phenotype. Further studies are needed to establish whether AC-EVOO could represent a new therapeutic target to prevent peritoneal fibrosis.

Monitoring the effects of iodine prophylaxis in the adult population of southern Italy with deficient and sufficient iodine intake levels: a cross-sectional, epidemiological study

I prophylaxis is the most effective strategy to eradicate I deficiency disorders, but it has been shown to affect the thyroid disease pattern. In this study, we assessed the frequency of thyroid disorders in an adult population living in two areas of southern Italy after implementing I prophylaxis. To this aim, a cross-sectional, population-based study including 489 subjects from an I-deficient rural and an I-sufficient urban area of southern Italy was conducted. Thyroid ultrasound was performed on all participants, and urine and blood samples were collected from each subject. The levels of thyroid-stimulating hormone (TSH), thyroglobulin (TgAb) and thyroperoxidase antibodies (TPOAb), urinary I excretion (UIE), and thyroid volume and echogenicity were evaluated. We found that the median UIE was higher in the urban than in the rural area (P=0·004), whereas the prevalence of subjects affected by goitre was higher in the rural compared with the urban area (P=0·003). Positive TgAb rather than TPOAb were more frequent in subjects from the urban area compared with the rural area (P=0·009). The hypoechoic pattern at thyroid ultrasound (HT-US) was similar between the two areas, but TgAb were significantly higher (P=0·01) in HT-US subjects from the urban area. The frequency of elevated TSH did not differ between the two screened populations, and no changes were found for TgAb positivity in subjects with high TSH in the urban compared with the rural area. Our findings support that the small risks of I supplementation are far outweighed by the substantial benefits of correcting I deficiency, although continued monitoring of populations is necessary.