Role of glioma stem cells in promoting tumor chemo- and radioresistance: A systematic review of potential targeted treatments

BACKGROUND Gliomas pose a significant challenge to effective treatment despite advancements in chemotherapy and radiotherapy.Glioma stem cells(GSCs),a subset within tumors,contribute to resistance,tumor heterogeneity,and plasticity.Recent studies reveal GSCs’role in therapeutic resistance,driven by DNA repair mechanisms and dynamic transitions between cellular states.Resistance mechanisms can involve different cellular pathways,most of which have been recently reported in the literature.Despite progress,targeted therapeutic approaches lack consensus due to GSCs’high plasticity.AIM To analyze targeted therapies against GSC-mediated resistance to radio-and chemotherapy in gliomas,focusing on underlying mechanisms.METHODS A systematic search was conducted across major medical databases(PubMed,Embase,and Cochrane Library)up to September 30,2023.The search strategy utilized relevant Medical Subject Heading terms and keywords related to including“glioma stem cells”,“radiotherapy”,“chemotherapy”,“resistance”,and“targeted therapies”.Studies included in this review were publications focusing on targeted therapies against the molecular mechanism of GSC-mediated re-sistance to radiotherapy resistance(RTR).RESULTS In a comprehensive review of 66 studies on stem cell therapies for SCI,452 papers were initially identified,with 203 chosen for full-text analysis.Among them,201 were deemed eligible after excluding 168 for various reasons.The temporal breakdown of studies illustrates this trend:2005-2010(33.3%),2011-2015(36.4%),and 2016-2022(30.3%).Key GSC models,particularly U87(33.3%),U251(15.2%),and T98G(15.2%),emerge as significant in research,reflecting their representativeness of glioma characteristics.Pathway analysis indicates a focus on phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(mTOR)(27.3%)and Notch(12.1%)pathways,suggesting their crucial roles in resistance development.Targeted molecules with mTOR(18.2%),CHK1/2(15.2%),and ATP binding cassette G2(12.1%)as frequent targets underscore their importance in overcoming GSC-mediated resistance.Various therapeutic agents,notably RNA inhibitor/short hairpin RNA(27.3%),inhibitors(e.g.,LY294002,NVP-BEZ235)(24.2%),and monoclonal antibodies(e.g.,cetuximab)(9.1%),demonstrate versatility in targeted therapies.among 20 studies(60.6%),the most common effect on the chemotherapy resistance response is a reduction in temozolomide resistance(51.5%),followed by reductions in carmustine resistance(9.1%)and doxorubicin resistance(3.0%),while resistance to RTR is reduced in 42.4%of studies.CONCLUSION GSCs play a complex role in mediating radioresistance and chemoresistance,emphasizing the necessity for precision therapies that consider the heterogeneity within the GSC population and the dynamic tumor microenvironment to enhance outcomes for glioblastoma patients.

Integration between Genomic and Computational Statistical Surveys for the Screening of SNP Genetic Variants in Inflammatory Bowel Disease (IBD) Pediatric Patients*

Inflammatory bowel diseases (IBD) are complex multifactorial disorders that include Crohn’s disease (CD) and ulcerative colitis (UC). Considering that IBD is a genetic and multifactorial disease, we screened for the distribution dynamism of IBD pathogenic genetic variants (single nucleotide polymorphisms;SNPs) and risk factors in four (4) IBD pediatric patients, by integrating both clinical exome sequencing and computational statistical approaches, aiming to categorize IBD patients in CD and UC phenotype. To this end, we first aligned genomic read sequences of these IBD patients to hg19 human genome by using bowtie 2 package. Next, we performed genetic variant calling analysis in terms of single nucleotide polymorphism (SNP) for genes covered by at least 20 read genomic sequences. Finally, we checked for biological and genomic functions of genes exhibiting statistically significant genetic variant (SNPs) by introducing Fitcon genomic parameter. Findings showed Fitcon parameter as normalizing IBD patient’s population variability, as well as inducing a relative good clustering between IBD patients in terms of CD and UC phenotypes. Genomic analysis revealed a random distribution of risk factors and as well pathogenic SNPs genetic variants in the four IBD patient’s genome, claiming to be involved in: i) Metabolic disorders, ii) Autoimmune deficiencies;iii) Crohn’s disease pathways. Integration of genomic and computational statistical analysis supported a relative genetic variability regarding IBD patient population by processing IBD pathogenic SNP genetic variants as opposite to IBD risk factor variants. Interestingly, findings clearly allowed categorizing IBD patients in CD and UC phenotypes by applying Fitcon parameter in selecting IBD pathogenic genetic variants. Considering as a whole, the study suggested the efficiency of integrating clinical exome sequencing and computational statistical tools as a right approach in discriminating IBD phenotypes as well as improving inflammatory bowel disease (IBD) molecular diagnostic process.

The Role of University in the Education of Professional Accountants： Degree of IES 2 Application

In the period following the beginning of the global economic crisis, the quality of the education provided to support the accounting profession was debated. Simultaneously, Education for Sustainable Development （ESD） is becoming increasingly important. The change in the accounting profession and in accounting education offers an opportunity to formulate initial observations on the Italian university world in terms of the preparation of future professional accountants. This study uses the International Education Standards （lESs） published by the International Accounting Education Standards Board （IAESB）, in particular the IES 2 （content of professional accounting education programs）. The aim of the paper, which is of an empirical nature, is to understand the role of universities in Italy in the preparation of accountants in the light of the IES 2 provisions. The study and its policy implications may contribute to the debate on accounting education at national and international levels, also in the light of ESD.

A molecular phenology scale of grape berry development

Fruit growth and development consist of a continuous succession of physical,biochemical,and physiological changes driven by a genetic program that dynamically responds to environmental cues.Establishing recognizable stages over the whole fruit lifetime represents a fundamental requirement for research and fruit crop cultivation.This is especially relevant in perennial crops like grapevine(Vitis vinifera L.)to scale the development of its fruit across genotypes and growing conditions.In this work,molecular-based information from several grape berry transcriptomic datasets was exploited to build a molecular phenology scale(MPhS)and to map the ontogenic development of the fruit.The proposed statistical pipeline consisted of an unsupervised learning procedure yielding an innovative combination of semiparametric,smoothing,and dimensionality reduction tools.The transcriptomic distance between fruit samples was precisely quantified by means of the MPhS that also enabled to highlight the complex dynamics of the transcriptional program over berry development through the calculation of the rate of variation of MPhS stages by time.The MPhS allowed the alignment of time-series fruit samples proving to be a complementary method for mapping the progression of grape berry development with higher detail compared to classic time-or phenotype-based approaches.

Dissecting the effect of soil on plant phenology and berry transcriptional plasticity in two Italian grapevine varieties (Vitis vinifera L.).

Grapevine embodies a fascinating species as regards phenotypic plasticity and genotype-per-environment interactions.The terroir,namely the set of agri-environmental factors to which a variety is subjected,can influence the phenotype at the physiological,molecular,and biochemical level,representing an important phenomenon connected to the typicality of productions.We investigated the determinants of plasticity by conducting a field-experiment where all terroir variables,except soil,were kept as constant as possible.We isolated the effect of soils collected from different areas,on phenology,physiology,and transcriptional responses of skin and flesh of a red and a white variety of great economic value:Corvina and Glera.Molecular results,together with physio-phenological parameters,suggest a specific effect of soil on grapevine plastic response,highlighting a higher transcriptional plasticity of Glera in respect to Corvina and a marked response of skin compared to flesh.Using a novel statistical approach,we identified clusters of plastic genes subjected to the specific influence of soil.These findings could represent an issue of applicative value,posing the basis for targeted agricultural practices to enhance the desired characteristics for any soil/cultivar combination,to improve vineyards management for a better resource usage and to valorize vineyards uniqueness maximizing the terroir-effect.

The medicinal chemistry of Urtica dioica L.:from preliminary evidence to clinical studies supporting its neuroprotective activity

Urtica dioica is a perennial herb from the family of Urticaceae that is commonly known as stinging nettle.This plant is widespread in Europe,Africa,America,and a part of Asia,as it adapts to different environments and climatic condi-tions.The leaves,stalk,and bark of U.dioica found applications in the field of nutrition,cosmetics,textile,pest control and pharmacology.In this connection,bioactive chemical constituents such as flavonoids,phenolic acids,amino acids,carotenoids,and fatty acids have been isolated from the plant.With this review,we aim at providing an updated and comprehensive overview of the contributions in literature reporting computational,in vitro,pre-clinical and clini-cal data supporting the therapeutic applications of U.dioica.Experimental evidence shows that U.dioica constituents and extracts can provide neuroprotective effects by acting through a combination of different molecular mecha-nisms,that are discussed in the review.These findings could lay the basis for the identification and design of more effective tools against neurodegenerative diseases.

Phosphodiesterase 9 localization in cytoplasm and nucleus:the gateway to selective targeting in neuroprotection?

The umbrella term"neurodege ne rative disorders"(NDDs) refers to several conditions characterized by a progressive loss of structure and function of cells belonging to the nervous system.Such diseases affect more than 50million people worldwide.Neurodegenerative disorders are characterized by sundry factors and pathophysiological mechanisms that a re challenging to be fully profiled.Many of these rely on cell signaling pathways to preserve homeostasis,involving second messengers such as cyclic adenosine monophosphate (cAMP)and cyclic guanosine 3',5'-monophosphate(cGMP).Their ability to control the duration and amplitude of the signaling cascade is given by the presence of several common and uncommon effectors.Protein kinases A and G (PKA and PKG),phosphodiesterases (PDEs),and scaffold proteins are among them.

Oncologic outcomes with and without amniotic membranes in robotic-assisted radical prostatectomy:A propensity score matched analysis

Objective:Placement of human placenta derived grafts during robotic-assisted radical prostatectomy(RARP)hastens the return of continence and potency.The long-term impact on the oncologic outcomes remains to be investigated.Our objective was to determine the oncologic outcomes of patients with dehydrated human amnion chorion membrane(dHACM)at RARP compared to a matched cohort.Methods:In a referral centre,from August 2013 to October 2019,599 patients used dHACM in bilateral nerve-sparing RARP.We excluded patients with less than 12 months follow-up,simple prostatectomy,and unilateral nerve-sparing.Patients with dHACM(amnio group)were 529,and were propensity score matched 1:1 to 2465 patients without dHACM(non-amnio group)and a minimum follow-up of 36 months.At the time of RARP,dHACM was placed around the neurovascular bundle in the amnio group.Continuous and categorical variables in matched groups was tested by two-sample Kolmogorov-Smirnov test and Fisher's exact test respectively.Outcomes measured were biochemical recurrence(BCR),adjuvant and salvage therapy rates.Results:Propensity score matching resulted in two groups of 444 patients.Cumulative incidence functions for BCR did not show a difference between the groups(p=0.3).Patients in the non-amnio group required salvage therapy more frequently than the amnio group,particularly after partial nerve-sparing RARP(6.3%vs.2.3%,p=0.001).Limitations are the absence of prospective randomization.Conclusion:The data suggest that using dHACM does not have a negative impact on BCR in patients.Outcomes of cancer specific and overall survival will require follow-up study to increase our understanding of these grafts’impact on prostate cancer biology.

Current status and future perspectives on stem cell transplantation for spinal cord injury

BACKGROUND Previous assessments of stem cell therapy for spinal cord injuries(SCI)have encountered challenges and constraints.Current research primarily emphasizes safety in early-phase clinical trials,while systematic reviews prioritize effectiveness,often overlooking safety and translational feasibility.This situation prompts inquiries regarding the readiness for clinical adoption.AIM To offer an up-to-date systematic literature review of clinical trial results concerning stem cell therapy for SCI.METHODS A systematic search was conducted across major medical databases[PubMed,Embase,Reference Citation Analysis(RCA),and Cochrane Library]up to October 14,2023.The search strategy utilized relevant Medical Subject Heading(MeSH)terms and keywords related to"spinal cord","injury","clinical trials","stem cells","functional outcomes",and"adverse events".Studies included in this review consisted of randomized controlled trials and non-randomized controlled trials reporting on the use of stem cell therapies for the treatment of SCI.RESULTS In a comprehensive review of 66 studies on stem cell therapies for SCI,496 papers were initially identified,with 237 chosen for full-text analysis.Among them,236 were deemed eligible after excluding 170 for various reasons.These studies encompassed 1086 patients with varying SCI levels,with cervical injuries being the most common(42.2%).Bone marrow stem cells were the predominant stem cell type used(71.1%),with various administration methods.Follow-up durations averaged around 84.4 months.The 32.7%of patients showed functional improvement from American spinal injury association Impairment Scale(AIS)A to B,40.8%from AIS A to C,5.3%from AIS A to D,and 2.1%from AIS B to C.Sensory improvements were observed in 30.9%of patients.A relatively small number of adverse events were recorded,including fever(15.1%),headaches(4.3%),muscle tension(3.1%),and dizziness(2.6%),highlighting the potential for SCI recovery with stem cell therapy.CONCLUSION In the realm of SCI treatment,stem cell-based therapies show promise,but clinical trials reveal potential adverse events and limitations,underscoring the need for meticulous optimization of transplantation conditions and parameters,caution against swift clinical implementation,a deeper understanding of SCI pathophysiology,and addressing ethical,tumorigenicity,immunogenicity,and immunotoxicity concerns before gradual and careful adoption in clinical practice.

indocyanine green-based fluorescence imaging in visceral and hepatobiliary and pancreatic surgery:State of the art and future directions

In recent years, the use of fluorescence-guided surgery(FGS) to treat benign and malignant visceral, hepatobiliary and pancreatic neoplasms has significantly increased. FGS relies on the fluorescence signal emitted by injected substances(fluorophores) after being illuminated by ad hoc laser sources to help guide the surgical procedure and provide the surgeon with real-time visualization of the fluorescent structures of interest that would be otherwise invisible. This review surveys and discusses the most common and emerging clinical applications of indocyanine green(ICG)-based fluorescence in visceral, hepatobiliary and pancreatic surgery. The analysis, findings, and discussion presented here rely on the authors' significant experience with this technique in their medical institutions, an up-to-date review of the most relevant articles published on this topic between 2014 and 2018, and lengthy discussions with key opinion leaders in the field during recent conferences and congresses. For each application, the benefits and limitations of this technique, as well as applicable future directions, are described. The imaging of fluorescence emitted by ICG is a simple, fast,relatively inexpensive, and harmless tool with numerous different applications in surgery for both neoplasms and benign pathologies of the visceral and hepatobiliary systems. The ever-increasing availability of visual systems that can utilize this tool will transform some of these applications into the standard of care in the near future. Further studies are needed to evaluate the strengths and weaknesses of each application of ICG-based fluorescence imaging in abdominal surgery.

MR with Gd-EOB-DTPA in assessment of liver nodules in cirrhotic patients

To date the imaging diagnosis of liver lesions is based mainly on the identification of vascular features, which are typical of overt hepatocellular carcinoma(HCC), but the hepatocarcinogenesis is a complex and multistep event during which, a spectrum of nodules develop within the liver parenchyma, including benign small and large regenerative nodule(RN), low-grade dysplastic nodule(LGDN), high-grade dysplastic nodule(HGDN), early HCC, and well differentiated HCC. These nodules may be characterised not only on the basis of their respective different blood supplies, but also on their different hepatocyte function. Recently, in liver imaging the introduction of hepatobiliary magnetic resonance imaging contrast agent offered the clinicians the possibility to obtain, at once, information not only related to the vascular changes of liver nodules but also information on hepatocyte function. For this reasons this new approach becomes the most relevant diagnostic clue for differentiating low-risk nodules(LGDN-RN) from highrisk nodules(HGDN/early HCC or overt HCC) and consequently new diagnostic algorithms for HCC have been proposed. The use of hepatobiliary contrast agents is constantly increasing and gradually changing the standard of diagnosis of HCC. The main purpose of this review is to underline the added value of Gd-EOB-DTPA in early-stage diagnoses of HCC. We also analyse the guidelines for the diagnosis and management of HCC, the key concepts of HCC development, growth and spread and the imaging appearance of precursor nodules that eventually may transform into overt HCC.

Multimodal treatment of gastric cancer in the west: Where are we going?

The incidence of gastric cancer(GC) is decreasing worldwide,especially for intestinal histotype of the distal third of the stomach.On the contrary,proximal location and diffuse Lauren histotype have been reported to be generally stable over time.In the west,no clear improvement in long-term results was observed in clinical and population-based studies.Results of treatment in these neoplasms are strictly dependent on tumor stage.Adequate surgery and extended lymphadenectomy are associated with good long-term outcome in early-stage cancer; however,results are still unsatisfactory for advanced stages(Ⅲ and Ⅳ),for which additional treatments could provide a survival benefit.This implies a tailored approach to GC.The aim of this review was to summarize the main multimodal treatment options in advanced resectable GC.Perioperative or postoperative treatments,including chemotherapy,chemoradiotherapy,targeted therapies,and hyperthermic intraperitoneal chemotherapy have been reviewed,and the main ongoing and completed trials have been analyzed.An original tailored multimodal approach to non-cardia GC has been also proposed.

Recent advances in non-invasive magnetic resonance imaging assessment of hepatocellular carcinoma

Magnetic resonance(MR) imaging of the liver is an important tool for the detection and characterization of focal liver lesions and for assessment of diffuse liver disease,having several intrinsic characteristics,represented by high soft tissue contrast,avoidance of ionizing radiation or iodinated contrast media,and more recently,by application of several functional imaging techniques(i.e.,diffusion-weighted sequences,hepatobiliary contrast agents,perfusion imaging,magnetic resonance(MR)-elastography,and radiomics analysis). MR functional imaging techniques are extensively used both in routine practice and in the field of clinical and pre-clinical research because,through a qualitative rather than quantitative approach,they can offer valuable information about tumor tissue and tissue architecture,cellular biomarkers related to the hepatocellular functions,or tissue vascularization profiles related to tumor and tissue biology. This kind of approach offers in vivo physiological parameters,capable of evaluating physiological and pathological modifications of tissues,by the analysis of quantitative data that could be used in tumor detection,characterization,treatmentselection,and follow-up,in addition to those obtained from standard morphological imaging. In this review we provide an overview of recent advanced techniques in MR for the diagnosis and staging of hepatocellular carcinoma,and their role in the assessment of response treatment evaluation.

Effects of artificial aging conditions on mechanical properties of gravity cast B356 aluminum alloy

The age hardening behavior of gravity cast B356 aluminum alloy was investigated by differential scanning calorimetry(DSC), hardness measurements and tensile tests. Three different artificial aging temperatures were selected, namely 155, 165 and 180 °C, with heat treatment time from 40 min to 32 h. DSC analysis results show that cluster formation begins below room temperature(at around-10 °C). Since cluster formation influences the subsequent precipitation of the main strengthening β'' phase, it can be inferred that a delay between solutionizing and artificial aging has a detrimental effect on the mechanical properties of the alloy. It was also confirmed that the hardness and the tensile properties of the alloy reach the maximum values when β'' phase is completely developed during the artificial aging. This happens after 16 h for samples aged at 155 °C, after 6 h for samples aged at 165 °C and after 4 h for samples aged at 180 °C. A subsequent decrease of the mechanical properties, observed only in the sample aged at the highest temperature, with increasing aging time can be associated with the transformation of the coherent β'' phase into the semi-coherent β' phase. Finally, the activation energy associated with the precipitation of β'' phase was calculated to be 57.2 k J/mol.

Decreased expression of Klotho in cardiac atria biopsy samples from patients at higher risk of atherosclerotic cardiovascular disease

Background Klotho proteins （α- and β） are membrane-based circulating proteins that regulate cell metabolism, as well as the lifespan modulating activity of Fibroblast Growth Factors （FGFs）. Recent data has shown that higher plasma circulating Klotho levels reduce cardio- vascular risk, suggesting Klotho has a protective role in cardiovascular diseases. However, although so far it has been identified in various organs, it is unknown whether cardiomyocytes express Klotho and FGFs, and whether high cardiovascular risk could affect cardiac expres- sion ofKlotho, FGFs and other molecules. Methods We selected 20 patients with an estimated 10-year high atherosclerotic cardiovascular disease and 10 age-matched control subjects with an estimated 10-year low risk undergone cardiac surgery for reasons other than coronary artery by-pass. In myocardial biopsies, we evaluated by immuno-histochemistry whether Klotho and FGFs were expressed in cardiomyo- cytes, and whether higher cardiovascular risk influenced the expression of other molecules involved in endoplasmic reticulum stress, oxida- tive stress, inflammation and fibrosis. Results Only cardiomyocytes of patients with a higher cardiovascular risk showed lower expression of Klotho, but higher expressions of FGFs. Furthermore, higher cardiovascular risk was associated with increased expression of oxidative and endoplasmic reticular stress, inflammation and fibrosis. Conclusions This study showed for the first time that Klotho proteins are ex- pressed in human cardiomyocytes and that cardiac expression of Klotho is down-regulated in higher cardiovascular risk patients, while expression of stress-related molecules were significantly increased.

Gadoxetic acid magnetic-enhanced resonance imaging in the diagnosis of cholangiocarcinoma

The use of liver magnetic resonance imaging is increasing thanks to itsmultiparametric sequences that allow a better tissue characterization, and the useof hepatobiliary contrast agents. This review aims to evaluate gadoxetic acidenhanced magnetic resonance imaging in the diagnosis and staging ofcholangiocarcinoma and its different clinical and radiological classificationsproposed in the literature. We also analyze the epidemiology, risk factors incorrelation with clinical findings and laboratory data.

Occasional finding of mesenteric lipodystrophy during laparoscopy:A difficult diagnosis

Mesenteric lipodystrophy is a rare pathological condition affecting the mesentery. Its initial presentation is typically asymptomatic. Pathological characteristics are unspecific, and generally attributed to inflammation, unless the diagnosis is suspected. Laparoscopy done for other reasons has been, as in this case, unsuccessful in providing evidence for the correct diagnosis, thus requiring laparotomy due to lack of diagnostic tissue. After 6 mo no further medical therapy is required, as the patient remains asymptomatic. Discussion of this case and a brief review of the literature are presented in the following paragraphs.

Optimization of heat treatment of gravity cast Sr-modified B356 aluminum alloy

In recent years,certain foundry processes have made it possible to obtain products with very thin parts,below the4mmthreshold of the permanent mold casting technology.The safety margins of these castings have been reduced,so the T6heattreatment conditions adopted for the Al?7Si?Mg alloys need to be investigated to identify the best combination of strength andductility.Furthermore,the cost and the production time associated with T6heat treatment have to be optimized.In the present work,an experimental study was carried out to optimize the solution treatment and artificial aging conditions in gravity cast thin bars ofB356aluminum alloy modified with Sr.Two solution temperatures were selected,530°C and550°C,respectively,with solutiontime ranging from2to8h,followed by water quenching and artificial aging at165°C with aging time from2to32h.The results ofhardness and tensile tests were correlated with differential scanning calorimetry(DSC)analysis.The best combination of mechanicalproperties and heat treatment duration was obtained with2h solutionizing at550°C and8h aging at165°C.DSC analysis showedthat the alloy’s mechanical properties reach the maximum value when theβ''phase is completely developed during the artificialaging.

Recurrence in node-negative advanced gastric cancer:Novel findings from an in-depth pathological analysis of prognostic factors from a multicentric series

AIM To analyze the clinicopathological characteristics of patients with both node-negative gastric carcinoma and diagnosis of recurrence during follow-up. METHODS We enrolled 41 patients treated with curative gastrectomy for p T2-4 a N0 gastric carcinoma between 1992 and 2010,who developed recurrence(Group 1). We retrospectively selected this group from the prospectively collected database of 4 centers belonging to the Italian Research Group for Gastric Cancer,and compared them with 437 p T2-4 a N0 patients without recurrence(Group 2). We analyzed lymphatic embolization,microvascular infiltration,perineural infiltration,and immunohistochemical determination of p53,Ki67,and HER2 in Group 1 and in a subgroup of Group 2(Group 2 bis) of 41 cases matched with Group 1 according to demographic and pathological characteristics. RESULTS T4 a stage and diffuse histotype were associated with recurrence in the group of p N0 patients. In-depth pathological analysis of two homogenous groups of p N0 patients,with and without recurrence during longterm follow-up(groups 1 and 2 bis),revealed two striking patterns: lymphatic embolization and perineural infiltration(two parameters that pathologists can easily report),and p53 and Ki67,represent significant factors for recurrence.CONCLUSION The reported pathological features should be considered predictive factors for recurrence and could be useful to stratify node-negative gastric cancer patients for adjuvant treatment and tailored follow-up.

Bsm I(rs1544410) and Fok I(rs2228570) vitamin D receptor polymorphisms, smoking, and body mass index as risk factors of cutaneous malignant melanoma in northeast Italy

Objective:To investigate whether vitamin D receptor gene(VDR)Bsm I-rs1544410 and Fok I-rs2228570 polymorphisms,smoking duration,and body mass index(BMI)are risk factors for cutaneous melanoma,especially metastatic melanoma.Methods:We studied 120 cutaneous melanoma cases[68 stage I and II non-metastatic melanoma(NMet M)patients,plus 52Stage III and IV metastatic melanoma(Met M)patients],and 120 matching healthy controls from northeast Italy.VDR polymorphisms were measured by restriction fragment length polymorphism analysis.Absence or presence of Bsm I and Fok I restriction sites was denoted by"B"and"F"or by"b"and"f,"respectively.Results:VDR-Bsm I bb genotype was more frequent among Met M(32.7%)than among NMet M cases(13.2%),with odds ratio(OR)=3.18.Comparison of all melanoma patients vs healthy controls showed that the following biomarkers were at risk:≥20 years of smoking(OR=2.43);≥20 years of smoking combined with bb(OR=4.78),Bb+bb(OR=2.30),Ff(OR=3.04),and Ff+ff(OR=3.08);obesity(BMI>30Conclusions:Risk factors for cutaneous Met M include two VDR polymorphisms combined with smoking duration and obesity.Results suggest gene-environment implications in melanoma susceptibility and severity.Future studies in larger cohorts and in subjects with different genetic background are warranted to extend our findings.