Metabolomic characterization of pigmented and non-pigmented potato cultivars using a joint and individual variation explained (JIVE)

Potatoes (Solanum tuberosum L.) are one of the most valuable agricultural crops, and the flesh of these tubers provides various classes of healthy compounds important for human nutrition. This work presents the results of a joint analysis of different chemical classes of compounds which provided insights on the metabolic characterization of pigmented and non-pigmented potato varieties collected from Italy. The identification of common or individual metabolic characteristics across the omic datasets (antioxidants, total polyphenolic content, polyphenols, and sugars) is conducted by Joint and Individual Variation Explained (JIVE), a data fusion multivariate approach. The common part of the multivariate model allowed the separation between non-pigmented and pigmented samples. Polyphenolic compounds were mainly responsible for the separation between purple-fleshed and red-skinned potatoes. An additional detailed analysis of the anthocyanin composition, including the acylated anthocyanins, allowed to pinpoint the diversities between the pigmented potato groups. Furthermore, the presence of an appreciable amount of hydroxycinnamic acids and anthocyanins in the purple-fleshed varieties, which are also characterized by a lower content of sugars, is found. Our results provide scientific evidence for the promotion of promising potato cultivars, which are characterized by a remarkable amount of various health benefit compound

Standardized Duplex Ultrasound-Based Protocol for Early Diagnosis of Transplant Renal Artery Stenosis: Results of a Single-Institution Retrospective Cohort Study

Transplant renal artery stenosis (TRAS) is the most frequent vascular complication after kidney transplantation (KT) and has been associated with potentially reversible refractory hypertension, graft dysfunction, and reduced patient survival. The aim of the study is to describe the outcomes of a standardized Duplex Ultrasound- (DU-) based screening protocol for early diagnosis of TRAS and for selection of patients potentially requiring endovascular intervention. We retrospectively reviewed our prospectively collected database of KT from January 1998 to select patients diagnosed with TRAS. The follow-up protocol was based on a risk-adapted, dynamic subdivision of eligible KT patients in different risk categories (RC) with different protocol strategies (PS). Of 598 patients included in the study, 52 (9%) patients had hemodynamically significant TRAS and underwent percutaneous angioplasty (PTA) and stent placement. Technical and clinical success rates were 97% and 90%, respectively. 7 cases of restenosis were recorded at follow-up and treated with re-PTA plus stenting. Both DU imaging and clinical parameters improved after stent placement. Prospective high-quality studies are needed to test the efficacy and safety of our protocol in larger series. Accurate trial design and standardized reporting of patient outcomes will be key to address the current clinical needs

Sviluppo di metodiche analitiche per la determinazione di composti di interesse biologico da matrici complesse di origine naturale

L’obiettivo generale del lavoro svolto in questa tesi di dottorato è stato quello di studiare tecniche cromatografiche di uso ormai consolidato, anche per applicazioni routinarie, in laboratori analitici, attraverso lo sviluppo di applicazioni su matrici complesse di tipo acquoso e lipidico, di origine vegetale o animale, per la determinazione simultanea dei componenti presenti al loro interno. Le tecniche cromatografiche studiate sono costituite da esempi di cromatografia liquida (HPLC-DAD, CE-DAD, TLC o HP-TLC) e da cromatografia gassosa (GC-FID). L’HPLC-DAD e la CE-DAD sono state applicate all’analisi di vitamine idrosolubili presenti in miscele di standard ed in preparati multivitaminici commerciali. L’HPLC è stata condotta su diversi materiali adsorbenti (HILIC, fase inversa alchilammidica, fase inversa RP-C18), mentre la CE è stata condotta attraverso due approcci, la CZE e la MEKC. Le metodiche sviluppate sono state applicate all’analisi di campioni di origine naturale a titolo incognito. Tramite HPLC, sia con l’approccio HILIC che in fase inversa C18, è stata effettuata l’analisi di lcuni tannini commerciali utilizzati sia come tannini enologici che come tannini da concia: questo ha consentito l’ottenimento di un fingerprinting per i campioni studiati. La tecnica TLC, nella variante a flusso forzato OPLC, ha permesso l’analisi di alcuni olii essenziali ed estratti di piante aromatiche ed il riconoscimento dei loro marcatori, ed ha consentito un miglioramento rispetto all’utilizzo della TLC classica per l’analisi dei campioni studiati. La tecnica HP-TLC, infine, è stata utilizzata anche per la determinazione delle principali famiglie di composti nel sebo umano; tramite utilizzo fotodensitometro è stato possibile ottenere una valutazione semiquantitativa dei composti presenti. La determinazione di alcuni marcatori in particolare è stata successivamente effettuata tramite GC-FID che ha permesso anche la valutazio dell’efficacia di alcuni supporti solidi adesivi per il campionamento del sebo

Comparison between TLC and OPLC fingerprints of commercial essential oils and ASE (Accelerated Solvent Extraction) plant extracts

Developing fit-to-purpose analytical screening methods for plant marker constituents by simple and affordable techniques represents a constant and important request from the current transformation chain of plant products. In this work, methods developed on classical thin-layer chromatography (TLC) and overpressured layer chromatography (OPLC) have been applied on some of the most valuable commercial plant products, such as star anise (Illicium verum Hook. f.) essential oil and extracts, common thyme (Thymus vulgaris L.) and sweet fennel (Foeniculum vulgare Mill.) essential oils and acerola fruit (Malpighia punicifolia L.) hydroalcoholic extracts, to evaluate OPLC potentiality in comparison with TLC performances in the detection of some characteristic constituents. OPLC provided higher performance with respect to TLC in all experiments performed due to higher selectivity demonstrated on all the tested marker compounds except for flavonoids in acerola extracts. Considering the usage of planar chromatography in the quality control of plant derivatives, the present paper shows how the OPLC protocols were both highly time- and solvent-saving in comparison with classical TLC

Assay of endogenous 3,5-diiodo-L-thyronine (3,5-T2) and 3,3’-diiodo-L-thyronine (3,3’-T2) in human serum.

Background: 3,5-diiodo-L-thyronine (3.5-T2) is an endogenous derivative of thyroid hormone with potential metabolic effects. It has been occasionally detect in human blood using High Performance Liquid Chromatography- tandem Mass Spectrometry (HPLC-MS/MS), but the results have been quite variable and quality control data were often missing. In this study we used an ad-hoc optimized Solid Phase Extraction (SPE)-based extraction method which allowed us to quantitate 3,5-T2 and its isomer 3,3’-T2 in human serum. Methods: Serum samples were obtained from 28 patients (8 healthy volunteers and 20 women undergoing endocrinological screening and found to be euthyroid). Two ml of serum were deproteinized with acetonitrile and then extracted using an SPE based process. To lower background noise, after extraction the samples were furtherly cleaned by hexane washing and acetonitrile precipitation of residual proteins. 3,5-T2 and 3,3’-T2 were analyzed by a Sciex API4000 mass spectrometer coupled to an Agilent Infinity 1290 LC system using isotope dilution method. In 20 patients total T3 and T4 were also assayed by HPLC-MS/MS. Results: Accuracy and precision for 3,5-T2 assay were 88-104% and 95-97%, respectively. Recovery, matrix effect and process efficiency averaged 78%, 108%, and 84% respectively. 3,5-T2 was detected in all samples and its concentration averaged (mean±SEM) 41±5 pg/ml, i.e. 78±9 pmol/l. In the same samples the concentration of 3,3’-T2 averaged 133±15 pg/ml, i.e. 253±29 pmol/l. 3,5-T2 concentration was significantly related to 3,3’-T2 concentration (r=0.540, P<0.01), while no significant correlation was observed with either T3 or T4 in the subset of patients in which these hormones were assayed. Conclusion: Our optimized extraction method is able to quantify 3,5-T2 and 3,3’-T2 in human serum. Their concentrations are in the subnanomolar range, and a significant correlation was detected between these two metabolites in healthy individuals

Assay of endogenous 3,5-diiodo-L-thyronine (3,5-T2) and 3,3’-diiodo-L-thyronine (3,3’-T2) in human serum: a feasibility study

3,5-diiodo-L-thyronine (3,5-T2) is an endogenous derivative of thyroid hormone with potential metabolic effects. It has been detected in human blood by immunological methods, but a reliable assay based on mass spectrometry (MS), which is now regarded as the gold standard in clinical chemistry, is not available yet. Therefore, we aimed at developing a novel ad-hoc optimized method to quantitate 3,5-T2 and its isomers by MS in human serum. Serum samples were obtained from 28 healthy subjects. Two ml of serum were deproteinized with acetonitrile and then subjected to an optimized solid phase extraction-based procedure. To lower background noise, the samples were furtherly cleaned by hexane washing and acetonitrile precipitation of residual proteins. 3,5-T2 and its isomers 3,3’-T2 and 3’,5’-T2 were then analyzed by HPLC coupled to tandem MS. Accuracy and precision for T2 assay were 88-104% and 95-97%, respectively. Recovery and matrix effect averaged 78% and +8%, respectively. 3,5-T2 was detected in all samples and its concentration averaged (mean±SEM) 41±5 pg/ml, i.e. 78±9 pmol/l. In the same samples the concentration of 3,3’-T2 averaged 133±15 pg/ml, i.e. 253±29 pmol/l, while 3’,5’-T2 was not detected. 3,5-T2 concentration was significantly related to 3,3’-T2 concentration (r=0.540, P<0.01), while no significant correlation was observed with either T3 or T4 in a subset of patients in which these hormones were assayed. In conclusion, our method is able to quantify 3,5-T2 and 3,3’-T2 in human serum. Their concentrations lie in the subnanomolar range, and a significant correlation was detected between these two metabolites in healthy individuals

Smart learning for urology residents during the COVID-19 pandemic and beyond: insights from a nationwide survey in Italy

The covid 19 pandemic represents a global emergency worldwide. Italy is one of the Countries that has been hit the most, facing since the beginning an urgent need to reshape the whole healthcare system to optimize resources

New-onset Chronic Kidney Disease After Surgery for Localised Renal Masses in Patients with Two Kidneys and Preserved Renal Function: A Contemporary Multicentre Study

Background: There is a lack of evidence on acute kidney injury (AKI) and new-onset chronic kidney disease (CKD) after surgery for localised renal masses (LRMs) in patients with two kidneys and preserved baseline renal function. Objective: To evaluate the prevalence and risk of AKI and new-onset clinically significant CKD (csCKD) in patients with a single renal mass and preserved renal function after being treated with partial (PN) or radical (RN) nephrectomy. Design, setting, and participants: We queried our prospectively maintained databases to identify patients with a preoperative estimated glomerular filtration rate (eGFR) of ≥60 ml/min/1.73 m2 and a normal contralateral kidney who underwent PN or RN for a single LRM (cT1-T2N0M0) between January 2015 and December 2021 at four high-volume academic institutions. Intervention: PN or RN. Outcome measurements and statistical analysis: The outcomes of this study were AKI at hospital discharge and the risk of new-onset csCKD, defined as eGFR <45 ml/min/1.73 m2, during the follow-up. Kaplan-Meier curves were used to examine csCKD-free survival according to tumour complexity. A Multivariable logistic regression analysis assessed the predictors of AKI, while a multivariable Cox regression analysis assessed the predictors of csCKD. Sensitivity analyses were performed in patients who underwent PN. Results and limitations: Overall, 2469/3076 (80%) patients met the inclusion criteria. At hospital discharge, 371/2469 (15%) developed AKI (8.7% vs 14% vs 31% in patients with low- vs intermediate- vs high-complexity tumours, p < 0.001). At the multivariable analysis, body mass index, history of hypertension, tumour complexity, and RN significantly predicted the occurrence of AKI. Among 1389 (56%) patients with complete follow-up data, 80 events of csCKD were recorded. The estimated csCKD-free survival rates were 97%, 93% and 86% at 12, 36, and 60 mo, respectively, with significant differences between patients with high- versus low-complexity and high- versus intermediate-complexity tumours (p = 0.014 and p = 0.038, respectively). At the Cox regression analysis, age-adjusted Charlson Comorbidity Index, preoperative eGFR, tumour complexity, and RN significantly predicted the risk of csCKD during the follow-up. The results were similar in the PN cohort. The main limitation of the study was the lack of data on eGFR trajectories within the 1st year after surgery and on long-term functional outcomes. Conclusions: The risk of AKI and de novo csCKD in elective patients with an LRM and preserved baseline renal function is not clinically negligible, especially in those with higher-complexity tumours. While baseline nonmodifiable patient/tumour-related characteristics modulate this risk, PN should be prioritised over RN to maximise nephron preservation if oncological outcomes are not jeopardised. Patient summary: In this study, we evaluated how many patients with a localised renal mass and two functioning kidneys, who were candidates for surgery at four referral European centres, experienced acute kidney injury at hospital discharge and significant renal functional impairment during the follow-up. We found that the risk of acute kidney injury and clinically significant chronic kidney disease in this patient population is not negligible, and was associated with specific baseline patient comorbidities, preoperative renal function, tumour anatomical complexity, and surgery-related factors, in particular the performance of radical nephrectomy