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Chronic kidney disease (CKD) represents a frequent comorbidity in cancer patients, especially for patients affected by urological cancers. Unfortunately, impaired kidney function may limit the choice of adequate oncological treatments for their potential nephrotoxicity or due to contraindications in case of a low glomerular filtration rate. For these patients, tailored nephrological and nutritional management is mandatory. The K-DIGO guidelines do not define whether the nutritional management of CKD could be useful also in CKD patients affected by urological cancer. In fact, in clinical practice, oncological patients often receive high-protein diets to avoid malnutrition. In our study, we investigated the nutritional and nephrological impact of a Mediterranean-like diet with a controlled protein intake (MCPD) on a cohort of 82 stage III-IV CKD patients. We compared two cohorts: one of 31 non-oncological CKD patients and the other of 51 oncological patients with CKD. The use of an MCPD had a favorable impact on both the oncological and non-oncological CKD patients with an amelioration in all the investigated parameters and with a better quality of life, with no cases of malnutrition or AKI.

BACKGROUND AND AIMS In daily clinical practice, an accurate assessment of renal function is of paramount importance in several categories of patients where the adequate medical or surgical treatment depends on the values of glomerular filtration rate (GFR). In particular, both oncological and urological patients require a personalized medical approach able to avoid misleading errors which could dramatically shorten their lifespan. Unfortunately, the most used method to measure GFR in these groups of patients is represented by the estimated glomerular filtration rate (eGFR), which harbours a significant error in comparison to gold standards methods (mGFR). The aim of this study was to determine the extent of the error of eGFR compared to the mGFR in a consecutive prospective cohort of oncological patients affected by urological malignancies. METHOD A total consecutive cohort of 352 patients enrolled in a single tertiary institution between 2018 and 2021 was collected in order to compare the most common eGFR formulas used by physicians (Cockroft-Gault, MDRD, CKD-EPI based on serum creatinine and/or serum cystatin and the new eGFR equation based on creatinine and cystatin without race adjustment) with the most widespread mGFR method (Iohexol Plasma Clearance). The study cohort was composed by 188 oncological patients affected by different types of urological malignancies (cases) before surgical operation and/or medical treatment and 164 non-oncological patients (controls) matched for baseline clinical variables and GFR. The agreement between eGFR and mGFR was evaluated using bias (as median of difference), precision (as interquartile range of difference—IQR) accuracy (as P30) and total deviation index (TDI). The differences between cohorts were evaluated with Fisher's exact test and Chi-squared test for ordinal characteristics and Wilcoxon rank sum test for continuous variables. Data analysis was performed using programming language R and Python. RESULTS Clinical data were as follows: median age 68 (IQR: 20.68), M/F ratio 3.19, median BMI 24.9 (IQR: 0.003, 24.938). A total of 61.2% of patients had hypertension, and 14.3% were diabetics. The median creatinine value in the overall population was 1.48 mg/dL (IQR: 0.45, 1.48); the median cystatin value was 1.26 mg/dL (IQR: 0.42, 1.26). Based on iohexol plasma clearance, 3.13% of patients were classified in CKD stage 1, 23.30% in stage 2, 29.55% in stage 3a, 27.27% in stage 3b, 14.77% in stage 4 and 1.99% in stage 5. The two matched cohorts of oncological and non-oncological patients displayed no statistical differences in terms of clinical variables and agreement parameters (TDI, CCC and P30). Surprisingly, both groups harboured a non-negligible error in each CKD class with a huge discrepancy between the eGFR formulas and the gold standard method (Figures 1 and 2), suggesting the great relevance of mGFR in the clinical decision-making algorithm, both in oncological and in non-oncological patients. CONCLUSION We observed that the error in the classification of CKD stages using eGFR formulas was very common, both in oncological and in nephrological patients, with a poor agreement with mGFR in all CKD classes. Therefore, mGFR remains a crucial tool for a correct clinical decision in all onconephrological patients who require surgery or potential nephrotoxic agents.

BACKGROUND AND AIMS Chronic kidney disease (CKD) represents the major postoperative long-term complication in patients who undergo radical nephrectomy (RN) for the presence of a renal mass. The development of a mild to severe grade of CKD can dramatically change the lifespan of this category of patients who can experience not only an augmented rate of cardiovascular diseases but also cancer recurrence and oncological therapies based on nephrotoxic agents. Therefore, preventive strategies for reducing the risk of CKD are strictly required. Among them, one of the most important tools is represented by the correct assessment of renal function. In fact, especially in the oncological scenario, there are various causes of factitious elevation or reduction of serum creatinine due to the frequent modification of body composition, measurement interference with the assay and altered tubular secretion. Nevertheless, the use of estimated GFR based on serum creatinine levels remains nowadays the most common method to define renal function in RN patients, both in the oncological and in the urological universe. The aim of this study was to determine the extent of the error of eGFR compared to the mGFR in a consecutive prospective cohort of RN patients who remained with a solitary kidney. METHOD A total consecutive cohort of 115 RN patients enrolled in a single tertiary institution between 2018 and 2021 was collected in order to compare the most common eGFR formulas used by physicians (Cockroft-Gault, MDRD, CKD-EPI based on serum creatinine and/or serum cystatin and the new eGFR equation based on creatinine and cystatin without race adjustment) with the most widespread mGFR method (Iohexol Plasma Clearance). The mGFR technique together with the serum creatinine/cystatin measurement was performed after 12 months from the operation in order to consider a steady state for the chronic renal function. All clinical variables were reported for each pt. CKD classification was defined accorded to K-DIGO 2012 guidelines. The agreement between eGFR and mGFR was evaluated using bias (as median of difference), precision (as interquartile range of difference-IQR), accuracy (as P30) and total deviation index (TDI). The differences between cohorts were evaluated with Fisher's exact test and Chi-squared test for ordinal characteristics and Wilcoxon rank sum test for continuous variables. Data analysis was performed using programming language R and Python. RESULTS Clinical data were as follows: median age 66 (IQR 27.66), M/F ratio 4.48, median BMI 24.6 (IQR: 0.003, 24.6). 51.3% of patients had hypertension, 10.4% were diabetics. The median creatinine level in the overall population was 1.49 mg/dL (IQR: 0.8, 1.49), the median cystatin level was 1.33 mg/dL (IQR: 0.55, 1.33). Based on iohexol plasma clearance, 0.87% patients were classified in CKD in stage 1, 17.39% in stage 2, 39.13% in stage 3a, 25.22% in stage 3b, 16.52% in stage 4 and 0.87% in stage 5. Surprisingly, a non-negligible error was reported in each CKD class with a huge discrepancy between the eGFR formulas and the gold standard method (Figures 1 and 2), suggesting the pivotal role of mGFR in the clinical decision-making algorithm for RN patients. CONCLUSION In daily clinical practice, an appropriate nephrological tailored follow-up based on mGFR is mandatory for RN patients with a solitary kidney. In fact, a simple renal evaluation using eGFR can only increase the risk of clinical errors, sometimes underestimating and sometimes overestimating the real renal function with important medical consequences.

BACKGROUND AND AIMS Acute kidney injury (AKI) is increasingly common and associated with adverse short- and long-term outcomes in oncological patients who require radical nephrectomy (RN) for the presence of renal cancer. As defined by the KDIGO Criteria classification system, the diagnosis of AKI is nowadays based on changes in the serum creatinine levels (sCr) or urine output. However, these parameters often underestimate the severity of renal damage and therefore recent insights have resulted in the implementation of proteinuria as means of more accurately assessing for AKI. Proteinuria represents indeed an indicator of both glomerular and renal endothelial injury in chronic disease (CKD), whereas in AKI the presence of proteinuria could be derived from a tubular dysfunction. In the RN scenario, it is still debatable if the development of a mild to severe proteinuria after surgery could be considered a marker of renal damage resulting in AKI or, on the opposite, could be a physiological sign of adaptive compensatory hyperfiltration promoted by the remanent counterlateral kidney. The aim of our study was to investigate if the development of a postoperative proteinuria plays a positive or negative role in the prevention of AKI in a consecutive cohort of nephrectomized patients affected by renal cancer. METHOD We enrolled a consecutive cohort of 131 patients who underwent RN due to the presence of a kidney mass suspected of malignancy from 2018 to 2021 in a tertiary care urological Institution. The following data were considered: age, gender, body mass index (BMI), smoke, TNM staging, Fuhrman grading, hypertension, diabetes, cardiovascular events, dyslipidaemia, medical therapy (ACE-inhibitors (ACEi), angiotensin II receptor blockers (ARBs), calcium antagonists, beta blockers and diuretics), proteinuria in 24 h, urine volume in 24 h, preoperative haematocrit and haemoglobin. Serum creatinine (s-Cr) values [Kinetic Picrate standardized (COBAS C 800) for IDMS] were collected before surgery (t 0) and at 24, 48 and 72 h after surgery and at dismissal (respectively t 1, t 2 and tf) to detect renal function fluctuations and the subsequent risk of AKI. GFR was estimated at baseline using CKD-EPI 2012 and MDRD formula. The level of proteinuria was measured at 24 h postoperative and was standardized on the urine volume. Each operated patient underwent the same pre-peri- and postoperative medical protocol in terms of surgical and anesthesiologic management. Patients affected by pre-existing medical nephropathies with proteinuria were excluded. Comparisons between groups were performed using the Kruskal–Wallis rank sum test for numerical variables and Pearson's Chi-squared test for categorical variables. Logistic regression was used to identify variables odds ratio for AKI onset after surgery. RESULTS Descriptive analysis is shown in Table 1. Our analysis surprisingly underlined that 66% of overall patients experienced AKI after surgery. Moreover, the analysis showed a significant difference (P value CONCLUSION Our work underlined that proteinuria does not represent a valid biomarker for AKI development after RN. On the contrary, basal eGFR, serum creatinine and postoperative urinary volume could be promising tools to predict and follow the acute renal damage after RN.

BACKGROUND AND AIMS Saffron is the dried stigmas of Crocus sativus L. Crocus sativus L belongs to the family of Iridaceae and is mainly cultivated in several countries with mild and dry climates. It is one of the most expensive spices in the world and the main reason for its great cost is that saffron is still cultivated and harvested as it has been for millennia by hand. Saffron's name is derived from the Arab word for yellow, a name reflecting the high concentration of carotenoid pigments present in the saffron flowers’ stigmas which contribute most to the colour profile of this spice. Saffron contains numerous bioactive molecules known for their antioxidant activities, but also some such as picrocrocin, crocetin and Safranal (Sfr), Crocin (Crc) which are known to possess the ability to inhibit tumoral growth by inducing apoptosis. The aim of this study is to understand whether these four saffron-derived bioactive molecules display tumoral growth inhibition activity on kidney and bladder cancer cell lines. METHOD Saffron samples used for this purpose possess both different origins (Tuscany and Lombardy) and harvesting years (T_2017, T_2018, L_2017, L_2018) and, in addition, Sfr and Crc standard samples were also tested. Identification of Crocins, Safranal, Picrocrocin and Flavonoids was carried out by HPLC/DAD and HPLC/MS analysis. The identity of polyphenols was ascertained using data from HPLC/DAD analyses by comparison and combination of their retention times and UV/Vis spectra with those of authentic standards and previously reported data (Vignolini et al., 2008). The extract characterization is summarized in the table below. The extracts’ effects on cancer cell viability were evaluated through MTT assays and the tumoral cell lines chosen were Caki-1 and 786-O for the kidney and RT112 and RT4 for the bladder. MDCK cells were used as a healthy kidney model. RESULTS In the presence of both pure Sfr and Crc, post-hoc tests have shown a time and concentration-dependent decrease in viability for all the cancerous models in analysis, with more visible results observable for Crc, while having no effect on MDCK viability. Being Crc is the most represented molecule in all the extracts, before performing MTT assays, a normalization of this compound's concentration was carried out. L_2017, which contained the highest Sfr concentration among all the extracts tested, showed the highest inhibiting activity. Hypothesizing that Sfr was the molecule responsible for the decrease in cancer cell viability, all extracts and Sfr and Crc standards were tested on RT112 cells after normalizing the contents of these two molecules for each condition. We observed that Crc and Sfr alone and the Sfr-Crc mix had a small impact on cancer cell viability when compared with the effects of whole extracts. CONCLUSION Whole extracts demonstrated a higher impact on cancer cell viability compared with the standards tested both singularly and combined. This study, therefore, recognizes that numerous molecules in combination with Sfr and Crc have a role in reducing tumoral cell viability, demonstrating that the phyto-complex may possess higher therapeutic effects.

BACKGROUND AND AIMS Nutritional therapy (NT) based on controlled protein intake represents a cornerstone in the management of chronic kidney disease (CKD). However, the international guidelines do not precisely define an adequate protein intake for onconephrological patients. The aim of our study is to investigate whether a low-normal protein high calorie (LNPHC) diet in a nephrologist-nutritionist combined approach (NNCA) and aimed at CKD patients can also be effective in improving the renal function in oncological patients which developed renal disease, while maintaining a high quality of life and a healthy nutritional status. METHOD A consecutive cohort of 113 patients was enrolled in the Urological Department at San Raffaele Scientific Institute between 2018 and 2021. Inclusion criteria were: age (>18 years), eGFR ( RESULTS The combined treatment produced eGFR and urea parameter improvements (49% of patients improved eGFR, 65% uremia) without negatively altering the anthropometrical outcomes. The nutritional status, as assessed by the MST, was preserved in both groups during the study. We notice specially an improvement in BMI (average deviation from ideal weight: 0.5 kg/h2 CS pts; 0.7 kg/h2 CT pts), an increase of PA [average difference (AD): 3.0°CS pts; 1.6°CT pts], BCMI (AD: 0.6 CS pts; 4.0 CT pts) and FFMI (AD: 0.1 female and 0.1 male for CS pts; 2.9 female and 1.1 male for CT pts) and a favourable decrease of WHC ratio (AD:–0.01 female and–0.02 male for CS pts; ––0.01 female and–0.01 male for CT pts), FM percentage (AD: –1.2 female and 1.2 male CS pts;–2.7 female and 2.9 male CT pts) and ECW/ICW (AD:–0.02 CS pts;–0.03 CT pts). Finally, RAPA test highlight that all patients considered themselves ameliorated after the diet period in term of lifestyle and Short-Form Health SF36 Questionnaire confirmed satisfaction for all eight domains of health including physical and mental health. CONCLUSION Our study suggests that LNPHC diet ameliorates the nephrological scenario, the metabolic complications and the nutritional perspective in uro-oncological CKD pts. Following NNCA, perceived quality of life has been pushed towards high scores and does not appear to be influenced by physical health and emotional status; however, there is a worsening in health change despite they describe their current health status as more than satisfactory. This may be explained by the fact that harsh clinical monitoring and food choice limitations may have increased the perception of the pathological condition, thereby increasing the sense of responsibility in adhering to the NT.

A reliable assessment of renal function is of paramount importance in several clinical assets in order to tailor a personalized medical approach. CKD classification system, created in 2002 by the National Kidney Foundation-sponsored Kidney Disease Outcomes Quality Initiative and then implemented in the following years by the K-DIGO guidelines, offered clinicians a new strategy to better identify nephrological patients at low or high risk to develop renal insufficiency, in order to avoid the progression to end-stage renal disease. However, the criteria used to create this classification did not consider some important aspects related to renal histology and glomerular filtration rate measurement, resulting in a possible over- or underestimation of the real established renal damage. In this mini-review, we will summarize the most relevant shortcomings in the CKD classifications, which can create misleading diagnosis in daily clinical practice.

The use of pembrolizumab has been largely accepted in several advanced types of cancers. PURE 01 study (NCT02736266) enrolled consecutively 143 patients with muscle-invasive bladder cancer who received 3 cycles of pembrolizumab 200 mg every 3 weeks before planned radical cystectomy (RC). Clinical, pathological and laboratory data were collected to investigate the relationship between renal function, immunotherapy and cancer-related outcomes. Serum creatinine and estimated glomerular filtration rate (eGFR) using Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) creatinine-equation 2009 were reported at baseline and after every cycle of pembrolizumab; the T stage from clinical classification TNM (cTNM) was stated before the treatment. Our analysis did not demonstrate a significant impairment of eGFR after any cycle of pembrolizumab, neither in the overall cohort nor in subgroups considering the T stages or the CKD G-categories according to K-DIGO 2012 classification. In conclusion, in neoadjuvant setting before RC our results suggest that pembrolizumab administration is safe for renal function preservation.

INTRODUCTION AND OBJECTIVE:Frailty is a key aspect, related to-but distinct from-comorbidities and age. Partial nephrectomy (PN) use has increased during the last decades. However, PN is a complex ...

Background and Aims Controlled low-normal protein (LNP) diet in CKD patient is effective in controlling the progression of renal impairment. Oncological guidelines explain how a high protein level should be always required in patients (pts) affected by malignancies in order to compensate the cancer derived metabolism and avoid catabolism. But what about the onco-nephrological pts? Aim of our study is to investigate the metabolic impact of a low-normal protein diet in a consecutive cohort of 103 nephrological pts affected or not by urological non-metastatic malignancies. Method A consecutive cohort of 103 pts was enrolled in the Urological Department at San Raffaele Scientific Institute between 2018-2020. Inclusion criteria were: Age (>18 years old), eGFR (< 90 ml/min/1.73), Malnutritional Screening Tool (MTS Results Population divided as follows: average age: 69.8 (+/- 10.3); ♂ : ♀ ratio: 2.4; Hypertension: 57.28%; Diabetes: 19,41%; CKD classification: 3,88% stage 2, 87,37% above stage 3. At time zero no pts were underweight and 51 % were overweight or obese with difference condition of FM percentage (FM % average: 24.3 % ♀ and 18.1 % ♂for CS pts vs 31.6 % ♀ and 18.2 % ♂for CT pts). Only 2.1% of pts had a PA of less than 4°, considered a negative prognostic index (PA average: 5.6° for CS pts vs 5.5° for CT pts) and 6.3 % of pts had a value of cell mass less than 8 kg/m2 indicative of reduced lean mass (BCMI average: 10.4 kg / m2 for CS pts vs 11 kg / m2 for CT pts). An increase in ECW/ICW ratio greater than 1 was more present in CT pts in respect to CS pts (41.6 % vs 22.8 % and ECW/ICW average: 1.01 for CT pts vs 0.93 for CS pts) as well as WC measurement associated with increased cardiovascular risk (WC average: 87 cm ♀ and 99 cm ♂ for CS pts vs 97 cm ♀ and 104 cm ♂ for CT pts). After 6 months of diet, we observed a similar behavior between the CS and the CT cohorts in terms of renal metabolites and eGFR profile. In fact, the 65% of onco-nephrological and the 55% of nephrological pts displayed a significative decrease in urea plasmatic levels (- 27,76 mg/dl) and eGFR improvement (+ 6,27 ml/min/1,73). The nutritional status, as assessed by the MST, was preserved in both groups during the study. In addition, all pts had an improvement in BMI (CS: 2.8 kg/m2; CT :1.3 kg/m2) PA (CS: 2.8°; CT: 1.3°), BCMI (CS: 1.31 kg / m2; CT :0.38 kg / m2) and FFMI (CS; 0.1 ♀ and 0.1 ♂; CT; 2.9 ♀ and 1.1 ♂) and a decrease of WC (CS: - 1.3 cm; CT– 1,65), FM percentage (CS: 0 %; CT – 2.7) and ECW/ICW (CS: 0.02; CT 0.03). Conclusion Our study suggests that LNP high calorie diet ameliorates the nephrological scenarios, the metabolic complications, and the nutritional perspective in uro-oncological pts with stage 2-5 CKD. A larger prospective study to validate these results is on-going.

Background and Aims Acute kidney injury (AKI) is a major postoperative complication in renal surgery for cancer, both in radical than in partial nephrectomy. One of the most intriguing arguments is to understand if normal renal function patients without renal urinary abnormalities can develop post-operative AKI after renal surgery. Aim of our study was to compare the AKI incidence in the two major renal surgeries approaches (radical and partial nephrectomies) in a selected cohort of patients with normal renal function. Method We performed a retrospective study of 216 patients who underwent radical (RN) or partial nephrectomy (PN) due to the presence of a kidney mass from 2000-2020 in a tertiary care Institution. Inclusion criteria: 1) Age> 18; 2) eGFR >;80 ml/min/1,73 using CKD-EPI formula 2012 3) absence of urinary abnormalities 4) presence of two kidneys at time operation. The following data were considered: age, gender, body mass index (BMI), TNM staging, hypertension, diabetes. Serum creatinine values were collected before surgery (t 0), at 48 hours after surgery and at dismissal. GFR was estimated at each time point using creatinine-based estimated glomerular filtration rate (eGFR) formula: CKD-EPI. We evaluated eGFR variation from the pre-surgical value to 48 h post and at dismissal. eGFR categories were created according to the KDIGO guidelines for G categories. Comparisons between groups were performed using Kruskal-Wallis ranks sum test for numerical variables and Pearson’s Chi square test for categorical variables. Logistic regression was used to identify variables ODDS Ratio for AKI onset after surgery. Results Clinical and pathological characteristics are reported in table 1. The cohort was composed by 216 patients with median Age 55 (IQR: 47, 64), M/F ratio 2.4, median BMI 25.8 (IQR: 23.3, 28.4), median eGFR 94.3 (IQR: 89.1, 101.6). According to CKD G classes, 79% were stage 1 and 2 1 were stage 2. The cohort was divided in two groups according to the type of surgery: 51% as RN and 49% as PN. Differences between the two groups were detected for Gender (Radical M/F ratio: 3.3; Partial M/F ratio: 1.8; p=0.4), basal eGFR (Radical Median:92.9 (88.1, 101.0); Partial Median:95.3 (90.7, 102.4); p=0.03) and CKD G class (Radical I:64%, II:36%; Partial I:78%, II:22%; p=0.02). Logistic regression for AKI onset showed as significant parameter (p Conclusion Our study highlights that both radical (55%) than partial (21%) nephrectomies harbor a non-negligible risk of post-operative AKI even in normal renal function patients without renal abnormalities. In addition, the impact of RN in normal renal function patients shows a dramatic incidence of AKI in the 50% of cases, suggesting that nephron sparing surgeries techniques should be always indicated to preserve renal function also in patients with an eGFR> 80 ml/min/1,73. A prospective comparison multicentric study with kidney living donor is on going.

Background and Aims Despite major advance in surgical techniques in the last decade, Chronic Kidney Disease (CKD) is still a major postoperative long-term complication in renal surgery for renal cancer, both in radical than in partial nephrectomy. However, it is still debatable if the renal hyperfiltration mechanism which happens after an acute loss of nephron mass could promote in oncological patients an effective vicariant process over time able to replace a new renal function comparable with the pre-operative one. Aim of our study was to compare the eGFR decay over time from pre-operative time surgery to 5 years follow up in the two major renal surgeries approaches (radical-RN- and partial nephrectomies- PN) in a selected cohort of patients with normal renal function at baseline. Method We performed a retrospective cohort-study of 271 consecutive patients who underwent radical (RN) or partial nephrectomy (PN) due to the presence of a kidney mass suspected of malignancy from 2000-2020 in a tertiary care urological Institution. Inclusion criteria were: 1) Age > 18 years old 2) eGFR > 80 ml/min/1,73 using CKD-EPI formula 2012 3) absence of urinary abnormalities . The following data were considered: age, gender, body mass index (BMI), TNM staging, hypertension, diabetes. Glomerular Filtration rate (GFR) was estimated at each time point using creatinine-based estimated glomerular filtration rate (eGFR) formula. We evaluated eGFR variation at the pre-surgical value, hospital dismissal, 6,12,24,36,48,60 months. eGFR categories were created according to the KDIGO guidelines for G categories in GFR setting different thresholds. Comparisons between groups were performed using Kruskal-Wallis ranks sum test for numerical variables and Pearson’s Chi square test for categorical variables. Logistic regression was used to identify variables ODDS Ratio for AKI onset after surgery. Results The study cohort was composed by 271 patients with median Age 56 (IQR: 48, 64), M/F ratio 2.3, median BMI 25.7 (IQR: 23.3, 28.3), median eGFR 94.3 (IQR: 89.1, 101.3) mL/min/1.73m^2. According to CKD G class, 70% were I and 30% were II. The cohort was divided in two groups according type of surgery: 44% as RN and 56% as PN. Differences between the two groups were detected for Gender (Radical M/F ratio: 3.2; Partial M/F ratio: 1.8; p=0.4), basal eGFR (Radical Median:92.3 (88.0, 99.8) mL/min/1.73m2; Partial Median:95.4 (90.1, 102.0) mL/min/1.73m2; p=0.01) and CKD G class (Radical I:64%, II:36%; Partial I:76%, II:24%; p=0.03). Two-way Anova for time variation of eGFR according other parameters, enlightened a significative difference for the type of surgery (p Conclusion Our study highlights that both RN than PN harbor a non negligible risk of post-operative CKD events even in normal renal function patients without renal abnormalities at 5 years from the operations. However, RN patients display a different behavior in term of renal compensation in respect of PN. In fact, RN pts tend to replace the acute loss of function derived from the absence of the contralateral kidney with an increase of eGFR , whereas PN pts tend to remain stable over time without any effort of hyperfiltration. A prospective comparison multicentric study with kidney living donor is on going.

Purpose: The role of non-tumour renal biopsy in predicting renal function after surgery for renal cell carcinoma (RCC) is poorly investigated. The aim of the study was to assess the impact of renal parenchymal histology on renal function after radical nephrectomy in a cohort of patients with RCC. Methods: This cohort study included 171 patients with RCC submitted to radical nephrectomy between 2006 and 2018. Two biopsy samples from normal parenchyma were collected at nephrectomy and renal parenchyma damage (RPD) was scored on histologic samples according to validated methodology. The outcomes were eGFR after surgery and its reduction > 25% relative to baseline at maximum 12months’ follow-up. Linear and logistic multivariable regression were used, adjusting for age at surgery, presence of hypertension, diabetes, clinical tumour size, time from surgery and basal eGFR. Results: 171 patients were enrolled and RPD was demonstrated in 64 (37%). Patients with RPD had more comorbidities (CCI > 2 in 25 vs. 9%, p < 0.001), in particular hypertension (70 vs. 53%; p = 0.03), diabetes with (5% vs. 0%, p = 0.007) or without (31 vs. 18%; p = 0.007) organ damage, cerebrovascular disease (19 vs. 5%; p = 0.006) and nephropathy (20 vs. 3%; p = 0.0004). At multivariable analyses, RPD was associated with lower eGFR (Est. −5.48; 95% CI −9.27: −1.7; p = 0.005) and with clinically significant reduction of eGFR after surgery (OR 3.06; 95% CI 1.17: 8.49; p = 0.026). Conclusions: Presence of RPD in non-tumour renal tissue is an independent predictor of functional impairment in patients with RCC. Such preliminary finding supports the use of parenchyma biopsy during clinical decision making.

Background and Aims Radical Nephrectomy is usually associated to the risk of future development of a mild to severe chronic kidney disease stage especially for those patients who already present early stages of CKD (e.g CKD class II and IIIa). Any insight on this topic could influence the clinical decision about the surgery. But how can we know for sure the magnitude of the renal function’s decay? In this preliminary work, our aim was to identify a new model able to predict at time surgery the renal function’s variation at 1 year from the operation Method We collected prospectively clinical data of a group of consecutive 114 patients who underwent radical nephrectomy (RN) for the presence of a benign or malignant renal mass. We estimated Glomerular Filtration Rate (eGFR) with MDRD formula. We considered the following clinical varibles: AKI onset (according to RIFLE criteria), age, gender, presence of blood hypertension, diabetes type II and BMI. Moreover, to investigate a possible correlation between renal basal histology and renal functional decay, renal biopsies were performed on each on the healthy part of the removed kidney > 3cm far from tumor. A pathological evaluation using a chronicity score (Remuzzi Score) was subsequently carried out evaluating damage on four parameters: (a) glomerular global sclerosis, (b) tubular atrophy, (c) interstitial fibrosis and (d) arterial narrowing. Statistical analysis were performed using generalized linear model (GLM), Kruskal-Wallis test and chi-square test. Multivariate analysis were applied using stepwise regressions method in order to select the best fitting model. Statistically significant correlations were considered for p-value Results At t0, 21% of the patients had an eGFR>90ml/min/1.73m2, 45% between 60 and 90, 23% between 30 and 45, and 11% under 45. Median observed decay after 12 months was 32.8% (IQR= 17.9%:41.9%).Taking in account the eGFR decay’s percentage there was a strong correlation with AKI onset (decay increased by 22.4%, CI= 14%:30.8%, p Conclusion A precise and reliable prediction of renal function decrease after RN represents a cornerstone for urologist and nephrologist in order to create a personalized medical approach and management.In our cohort of study, CKD stage I and II patients displayed a huge decrease of eGFR in respect to CKD stages III-IV over time. One possible biological explanation can be that the healthy kidney of the patients affected by moderate and severe CKD starts working with a compensatory mechanism before the entire removal of the kidney with cancer so that the surgical acute nephron loss does not represent a shock in comparison to healthy patients with an eGFR >90 ml/min. Our preliminary study identified a new clinical and pathological panel of variables able to predict at time zero the magnitude of eGFR decay after 1 year from surgical operation. Further studies are needed in order to validate and improve this model.

Background and Aims The use of Pembrolizumab, a selective humanized IgG4 kappa monoclonal antibody that inhibits the programmed death-1 receptor, has been largely accepted, especially in a series of trials in patients with advanced melanoma and in other cancers such as renal cell carcinoma (RCC), lymphoma and others. However, few recent studies demonstrated that Pembrolizumab promotes several renal dysfuncion such as acute tubular injury, acute interstitial nephritis and minimal change disease. To better elucidate this topic, we analyzed the renal function’s aspects from the PURE-01 protocol’s data, a phase 2, open label study of neoadjuvant Pembrolizumab before radical cystectomy (RC) for muscle invasive urothelial bladder cancer (MIUB). Method PURE-01 study (NCT02736266) enrolled preliminary 143 patients (pts). Eligibility criteria included: T2-T4N0 stage and residual disease after transurethral reduction of bladder. Pts received 3 cycles of Pembrolizumab 200 mg 3 weekly before RC. Computed tomography scan, FDG-PET/CT scan, and bladder multipara metric magnetic resonance imaging were performed during screening and before RC. Radiologically non-responders to pembro (per investigator decision) are given 3 additional courses of dose-dense MVAC chemotherapy. The collected data consisted in: the serum creatinine at baseline, after the first, second and last cycle in order to obtain the renal function by using eGFR formula (CKD-EPI-2009) for each time step; urine test analysis data after each cycle; the T stage from cTNM (AJCC TNM system-2019) before the treatment; clinical data such as presence of Diabetes, Hypertension, Hydronephrosis, BMI. The main outcome of the study was to correlate the renal function variation (from the pre-treatment to each treatment cycle) with the other clinical variables. Data analysis were performed using linear model, Kruskal-Wallis test and Wilcoxon test with holm’s correction. Results The median age of the patients was 68 years (62-73), with the 13% of the pts as female. We observed the presence of Hypertension, Hydronephrosis, Diabetes, Overweight (BMI≥25 and BMI Conclusion We observed that the treatment with neoadjuvant pembrolizumab, in patients with bladder cancer, does not correlate with a decay of eGFR, but on the contrary, from our data, is associated with a small improvement of renal function, especially for those patients who presented higher stage of T and hydronephrosis before the treatment suggesting the Permbrolizumab’s safety in neoadjuvant therapy before RC for MIUBC from the nephrological point of view. This results needs more in-depths studies to be confirmed.

The endocannabinoid system is a signaling system involved in a wide range of biological effects. Literature strongly suggests the endocannabinoid system role in the pathogenesis of cancer and that its pharmacological activation produces therapeutic benefits. Last research promotes the endocannabinoid system modulation by inhibition of endocannabinoids hydrolytic enzymes instead of direct activation of endocannabinoid receptors to avoid detrimental effects on cognition and motor control. Here we report the identification of N-acylethanolamine-hydrolyzing acid amidase (NAAA) inhibitors able to reduce cell proliferation and migration and cause cell death on different bladder cancer cell lines. These molecules were designed, synthesized and characterized and active compounds were selected by a fluorescence high-throughput screening method set-up on human recombinant NAAA that also allows to characterize the mechanism of inhibition. Together our results suggest an important role for NAAA in cell migration and in inducing tumor cell death promoting this enzyme as pharmacological target against bladder cancer.

Pasta is one of the basic foods of the Mediterranean diet and for this reason it was chosen for this study to evaluate its antioxidant properties. Three types of pasta were selected: buckwheat, rye and egg pasta. Qualitative–quantitative characterization analyses were carried out by HPLC-DAD to identify antioxidant compounds. The data showed the presence of carotenoids such as lutein and polyphenols such as indoleacetic acid, (carotenoids from 0.08 to 0.16 mg/100 g, polyphenols from 3.7 to 7.4 mg/100 g). To assess the effect of the detected metabolites, in vitro experimentation was carried out on kidney cells models: HEK-293 and MDCK. Standards of β-carotene, indoleacetic acid and caffeic acid, hydroalcoholic and carotenoid-enriched extracts from samples of pasta were tested in presence of antioxidant agent to determine viability variations. β-carotene and indoleacetic acid standards exerted a protective effect on HEK-293 cells while no effect was detected on MDCK. The concentrations tested are likely in the range of those reached in body after the consumption of a standard pasta meal. Carotenoid-enriched extracts and hydroalcoholic extracts showed different effects, observing rescues for rye pasta hydroalcoholic extract and buckwheat pasta carotenoid-enriched extract, while egg pasta showed milder dose depending effects assuming pro-oxidant behavior at high concentrations. The preliminary results suggest behaviors to be traced back to the whole phytocomplexes respect to single molecules and need further investigations.

Mesenchymal stem cells (MSC) as vehicles of therapeutic genes represent a unique tool to activate drugs within a neoplastic mass due to their property to home and engraft into tumours. In particular, MSC expressing the cytosine deaminase::uracil phosphoribosyltransferase (CD-MSC) have been previously demonstrated to inhibit growth of subcutaneous prostate cancer xenografts thanks to their ability to convert the non-toxic 5-fluorocytosine into the antineoplastic 5-fluorouracil. Since both the immune system and the tumour microenvironment play a crucial role in directing cancer progression, in order to advance towards clinical applications, we tested the therapeutic potential of this approach on animal models that develop autochthonous prostate cancer and preserve an intact immune system. As cell vectors, we employed adipose-tissue and bone-marrow MSC. CD-MSC toxicity on murine prostate cancer cells and tumour tropism were verified in vitro and ex-vivo before starting the preclinical studies. Magnetic Resonance Imaging was utilised to follow orthotopic tumour progression. We demonstrated that intravenous injections of CD-MSC cells, followed by intraperitoneal administration of 5-fluorocytosine, caused tumour regression in the transgenic adenocarcinoma of the mouse prostate (TRAMP) model, which develops aggressive and spontaneous prostate cancer. These results add new insights to the therapeutic potential of specifically engineered MSC in prostate cancer disease.

AIMS:To study micturition and bladder overactivity in female rats after chronic treatment with the fatty acid amide hydrolase (FAAH) inhibitor oleoyl ethyl amide (OEtA).METHODS:Sprague-Dawley rats ...

e18809Background: An accurate assessment of renal function is mandatory in the majority or urological and oncological patients to prevent renal impairment and cancer non-related deaths. Nowadays, t...

The transient receptor potential melastin 8 ion channel (TRPM8) is implicated in bladder sensing but limited information on TRPM8 antagonists in bladder overactivity is available. This study characterizes a new TRPM8-selective antagonist (DFL23448 [5-(2-ethyl-2H-tetrazol-5-yl)-2-(3-fluorophenyl)-1,3-thiazol-4-ol]) and evaluates it in cold-induced behavioral tests and tests on bladder function and experimental bladder overactivity in vivo in rats. DFL23448 displayed IC50 values of 10 and 21 nM in hTRPM8 human embryonic kidney 293 cells activated by Cooling Agent 10 or cold, but it had limited activity (IC50 > 10 μM) at transient receptor potential vanilloids TRPV1, TRPA1, or TRPV4 or at various G protein-coupled receptors. In rats, DFL23448 administered intravenously or orally had a half-life of 37 minutes or 4.9 hours, respectively. DLF23448 (10 mg/kg i.v.) reduced icilin-induced "wet dog-like" shakes in rats. Intravesical DFL23448 (10 mg/l), but not vehicle, increased micturition intervals, micturition volume, and bladder capacity. During bladder overactivity by intravesical prostaglandin E2 (PGE2), vehicle controls exhibited reductions in micturition intervals, micturition volumes, and bladder capacity by 37%-39%, whereas the same parameters only decreased by 12%-15% (P < 0.05-0.01 versus vehicle) in DFL23448-treated rats. In vehicle-treated rats, but not in DFL23448-treated rats, intravesical PGE2 increased bladder pressures. Intravenous DFL23448 at 10 mg/kg, but not 1 mg/kg DFL23448 or vehicle, increased micturition intervals, micturition volumes, and bladder capacity. During bladder overactivity by intravesical PGE2, micturition intervals, micturition volumes, and bladder capacity decreased in vehicle- and 1 mg/kg DFL23448-treated rats, but not in 10 mg/kg DFL23448-treated rats. Bladder pressures increased less in rats treated with DFL23448 10 mg/kg than in vehicle- or 1 mg/kg DFL23448-treated rats. DFL23448 (10 mg/kg i.v.), but not vehicle, prevented cold stress-induced bladder overactivity. Our results support a role for bladder TRPM8-mediated signals in experimental bladder overactivity.

Objective This study sought to develop a novel animal model to study the impact of nerve-sparing radical hysterectomy (NSRH) on female genital blood flow. Design In vivo animal study. Population Thirty Sprague-Dawley female rats. Materials and methods Female rats underwent either unilateral pelvic nerve (PN) crush (PNC; n = 9), or crush of both the PNs and all efferent nerves in the pelvic plexus (‘clock-nerve crush’, CNC; n = 9). Under anaesthesia, we electrically stimulated the crushed PN at 3 and 10 days after crush while monitoring blood pressure and recording clitoral and vaginal blood flows by laser Doppler. Uninjured PNs were stimulated as an internal control. Twelve additional rats were assigned either to bilateral PNC or sham surgery, and genital tissues were processed 10 days after injury for in vitro analysis. Main outcome measures Genital blood flow, nNOS, eNOS, collagen I-III. Results Stimulation of the crushed PN in both groups subjected to PNC and CNC induced significantly lower peak genital blood flow at 3 and 10 days (P

To study micturition and bladder overactivity in female rats after chronic treatment with the fatty acid amide hydrolase (FAAH) inhibitor oleoyl ethyl amide (OEtA).Sprague-Dawley rats received daily subcutaneous injections of OEtA (0.3 mg/kg), or vehicle for 2 weeks. Cystometries, organ bath studies, Western blot, and immunofluorescence were then used. Expressions of FAAH, cannabinoid 1 and 2 receptors (CB1 and CB2), mitogen-activated protein kinase (MAPK), vesicular acetyl choline-transporter protein (VAChT), and calcitonin gene-related peptide (CGRP) were evaluated.At baseline, OEtA-treated rats had higher values (P 0.05) of micturition intervals (MI) and volumes (MV), bladder capacity (BC), threshold pressure, and flow pressure than vehicle controls. Intravesical PGE2 reduced MI, MV, and BC, and increased basal pressure and the area under the curve in all rats. However, these urodynamic parameters were altered less by intravesical PGE2 in OEtA-treated rats (P 0.05 vs. vehicle controls). Compared to vehicle controls, detrusor from OEtA-treated rats had larger contractions to carbachol at 10-0.1 µM, but no difference in Emax was recorded. FAAH, CB1, CB2, VAChT, or CGRP was similarly expressed in bladders from all rats. In separate experiments, intravesical OEtA increased mucosal expression of phosphorylated MAPK.Chronic FAAH inhibition altered sensory urodynamic parameters and reduced bladder overactivity. Even if it cannot be excluded that OEtA may act on central nervous sensory pathways to contribute to these effects, the presence of FAAH and CB receptors in the bladder and activation of intracellular signals for CB receptors by intravesical OEtA suggest a local role for FAAH in micturition control.

INTRODUCTION AND OBJECTIVES: The endocannabinoid system modulates afferent signals under certain conditions. It is not known if the endocannabinoid-regulatory enzyme fatty acid amide hydrolase (FAAH) is located to or has a function in the ejaculatory system. We aimed to examine the expression of FAAH in the seminal vesicle and vas deferens and effects of inhibition of FAAH on seminal vesicle function and ejaculation. METHODS: After ethical approval, anesthetized male Sprague Dawley rats (250gram) were used for (I) recordings of intraluminal pressure of the seminal vesicle, or (II) pressures of the corpus spongiousum during apomorphine-induced (150 g/kg, s.c.) ejaculation. Effects by intraperitoneal oleoyl ethyl amide (OEtA; a FAAH inhibitor; 0.1, 1 and 10 mg/kg) or vehicle were studied. PCR, Western Blot and immunohistochemistry were used for studies of FAAH expression in rat and human tissues. Effect by OEtA (0.1-100 M) on isolated seminal vesicle was studied in organ baths. ANOVA and T-test were used for comparisons. RESULTS: PCR and WB demonstrated FAAH in rat and human seminal vesicles and vas deferens. FAAH was located in epithelial cells of both tissues. In human samples, the FAAH-2 isoform was similarly expressed. In isolated tissues, maximum inhibition by OEtA (100 M) on nerve-induced contractions were 21 9% (human;n 5) and 22 9% (rat;n 7). In vivo, the frequencies of spontaneous seminal vesicle contractions / min. were 1.08 0.32 (baseline) and 0.58 0.13 (OEtA 10mg/kg;p 0.05), and 1.23 0.41 (baseline) and 1.45 0.52 (vehicle; ns). OEtA increased basal pressure by 34 13% (vs. 0.3 2.9% for vehicle; p 0.05). Apomorphine-induced ejaculations were registered as suprasystolic intraspongious pressure spikes with striated muscle contractions and semen-expulsion from the urethra. Latencies to the first, second and third ejaculation were 6.0 1.2, 13.1 1.9, and 19.4 2.2 min. for vehicle (n 7); 12.7 3.2 (p 0.07), 19.6 4, and 22.8 2.6 min; and 21.8 3.3 (p 0.05) for OEtA (0.1 mg/kg; n 8), 26.9 5.1 (p 0.05), and 38.7 9.5 (p 0.05) min. for OEtA (1mg/kg; n 6). No differences in the number or pressure amplitudes of ejaculations were noted for OEtA and vehicle. CONCLUSIONS: FAAH is expressed in the rat and human vas deferens and seminal vesicle. FAAH inhibition seems to depress seminal vesicle excitability and prolongs latency for apomorphine-induced ejaculation in rats. Pharmacological targeting of FAAH may be an interesting principle to further develop in premature ejaculation.

The grey-lethal mouse (gl/gl) mutant most closely resembles the severe human malignant autosomal recessive OSTM1-dependent form of osteopetrosis that it has been described to be associated with neurological abnormalities. For this reason, we have analyzed the brain lipid composition (gangliosides, neutral glycosphingolipids, phospholipids and cholesterol), from gl/gl mice at different ages of development and compared with wild type mice. Both cholesterol and glycerophospholipid content and pattern in the gl/gl and control mice were very similar. In contrast, significant differences were observed in the content of several sphingolipids. Higher amount of the monosialogangliosides GM2 and GM3, and lower content of sphingomyelin, sulfatide and galactosylceramide were observed in the gl/gl brain with respect to controls. The low content of sphingomyelin, sulfatide and galactosylceramide is consistent with the immunohistochemical results showing that in the grey-lethal brain significant depletion and disorganization of the myelinated fibres is present, thus supporting the hypothesis that loss of function of the OSTM1 causes neuronal impairment and myelin deficit.

We studied the changes occurring in the membrane environment of prion protein (PrP) during apoptosis induced by low potassium in primary rat cerebellar neurons. Ceramide levels increased during apoptosis-inducing treatment, being doubled with respect to time-matched controls after 24 h. Sphingomyelin levels were parallely decreased, while cholesterol and ganglioside contents were not affected. Changes in ceramide and sphingomyelin composition were exclusively restricted to a detergent-resistant membrane fraction. The pro-apoptotic treatment was accompanied by the down-regulation of PrP and of the non-receptor kinase Fyn. The levels of PrP and Fyn were correspondingly reduced in the detergent-resistant membrane fraction. In control cells, the membrane microenvironment separated by immunoprecipitation with anti-PrP antibody contained 80% of the detergent-resistant PrP and 35% and 38% of the sphingolipids and cholesterol respectively. Upon low potassium treatment, 20% of the PrP originally present in the detergent-resistant fraction was immunoprecipitated, together with 19% of sphingolipids and 22% of cholesterol. Thus, PrP in the immunoprecipitate from apoptotic cells was ninefold less than in control ones, while sphingolipids and cholesterol were about 50% with respect to controls cells. The molar ratio between cholesterol, sphingomyelin and ceramide was 15 : 6 : 1 in the PrP-rich environment from control neurons, and 6 : 2 : 1 in that from apoptotic cells.

4-Oxo-N-(4-hydroxyphenyl)retinamide (4-oxo-4-HPR) is a recently identified metabolite of fenretinide (4-HPR). We explored the effectiveness of 4-oxo-4-HPR in inducing cell growth inhibition in ovarian, breast, and neuroblastoma tumor cell lines; moreover, we investigated the molecular events mediating this effect in two ovarian carcinoma cell lines, one sensitive (A2780) and one resistant (A2780/HPR) to 4-HPR. 4-Oxo-4-HPR was two to four times more effective than 4-HPR in most cell lines, was effective in both 4-HPR–sensitive and 4-HPR–resistant cells, and, in combination with 4-HPR, caused a synergistic effect. The tumor growth-inhibitory effects of 4-oxo-4-HPR seem to be independent of nuclear retinoid receptors (RAR), as indicated by the failure of RAR antagonists to inhibit its effects and by its poor ability to bind and transactivate RARs. Unlike 4-HPR, which only slightly affected the G1 phase of the cell cycle, 4-oxo-4-HPR caused a marked accumulation of cells in G2-M. This effect was associated with a reduction in the expression of regulatory proteins of G2-M (cyclin-dependent kinase 1 and cdc25c) and S (cyclin A) phases, and with an increase in the expression of apoptosis-related proteins, such as p53 and p21. Apoptosis was induced by 4-oxo-4-HPR in both 4-HPR–sensitive and 4-HPR–resistant cells and involved activation of caspase-3 and caspase-9 but not caspase-8. We also showed that 4-oxo-4-HPR, similarly to 4-HPR, increased reactive oxygen species generation and ceramide levels by de novo synthesis. In conclusion, 4-oxo-4-HPR is an effective 4-HPR metabolite that might act as therapeutic agent per se and, when combined with 4-HPR, might improve 4-HPR activity or overcome 4-HPR resistance. (Cancer Res 2006; 66(6): 3238-47)

We studied the membrane environment of cellular prion protein in primary cultured rat cerebellar neurons differentiated in vitro. In these cells, about 45% of total cellular prion protein (corresponding to a 35-fold enrichment) is associated with a low-density, sphingolipid- and cholesterol-enriched membrane fraction, that can be separated by flotation on sucrose gradient. Biotinylation experiments indicated that almost all prion protein recovered in this fraction was exposed at the cell surface. Prion protein was efficiently separated from this fraction by a monoclonal antibody immuno-separation procedure. Under conditions designed to preserve lipid-mediated membrane organization, several proteins were found in the prion protein-enriched membrane domains (i.e. the non-receptor tyrosine kinases Lyn and Fyn and the neuronal glycosylphosphatidylinositol-anchored protein Thy-1). The prion protein-rich membrane domains contained, as well, about 50% of the sphingolipids, cholesterol and phosphatidylcholine present in the sphingolipid-enriched membrane fraction. All main sphingolipids, including sphingomyelin, neutral glycosphingolipids and gangliosides, were similarly enriched in the prion protein-rich membrane domains. Thus, prion protein plasma membrane environment in differentiated neurons resulted to be a complex entity, whose integrity requires a network of lipid-mediated non-covalent interactions.