Metformin and vitamin D modulate adipose-derived stem cell differentiation towards the beige phenotype.

Adipose-derived stem cells (ADSCs) represent an ideal stem cell population for regenerative medicine. ADSC adipogenic differentiation is controlled by the activation of a specific transcriptional program, including epigenetic factors and key adipogenic genes. Under certain conditioned media, ADSCs can differentiate into several phenotypes. We previously demonstrated that bioactive molecules could counteract lipid accumulation and regulate adipogenesis, acting on inflammation and vitamin D metabolism. In the present paper, we aimed at evaluating the effect of metformin and vitamin D in targeting ADSC differentiation towards an intermediate phenotype, as beige adipocytes. We exposed ADSCs to different conditioned media and then we evaluated the levels of expression of main markers of adipogenesis, aP2, LPL and ACOT2. We also analysed the gene and protein expression of thermogenic UCP1 protein, and the expression of PARP1 and the beige specific marker TMEM26. Our results showed a novel effect of metformin and vitamin D not only in inhibiting adipogenesis, but also in inducing a specific 'brown-like' phenotype. These findings pave the way for their possible application in the control of de novo lipogenesis useful for the prevention of obesity and its related metabolic disorders.

Inflammatory Indexes as Predictive Biomarkers of Postoperative Complications in Oncological Thoracic Surgery.

The role of inflammatory responses in predicting outcomes in oncological thoracic surgery is still unclear. The aim of this study was to evaluate a series of blood count inflammation indexes as predicting factors for postoperative complications. We retrospectively studied 249 patients undergoing elective thoracic surgery in our institution between 2008 and 2020. A total of 184 patients underwent open surgery, and 65 underwent VATS. The neutrophil-to-lymphocyte (NLR), monocyte-to-lymphocyte (MLR), and platelet-to-lymphocyte (PLR) ratios, Systemic Inflammation Response Index (SIRI) were calculated preoperatively and on the first and fourth postoperative days, as well as a new derivative index, the Aggregate Inflammation Systemic Index (AISI). Univariate correlations evidenced a statistically significant association between the NLR at the fourth postoperative day and the occurrence of surgical complications in the global cohort (rho = 0.15, p = 0.03). A similar significant association with MLR on the fourth postoperative day is found in the open group (rho = -0.15, p = 0.048). NLR and LMR on the fourth postoperative day are associated with postoperative complications in the whole and open groups, respectively. Simple, easy-to-perform and inexpensive, blood cell count indexes may be useful in predicting complications in oncological thoracic surgery. A greater number of broader, prospective, randomized studies are necessary to confirm these findings.

Metformin and Vitamin D Modulate Inflammation and Autophagy during Adipose-Derived Stem Cell Differentiation.

Adipose-derived stem cells (ADSCs) came out from the regenerative medicine landscape for their ability to differentiate into several phenotypes, contributing to tissue regeneration both in vitro and in vivo. Dysregulation in stem cell recruitment and differentiation during adipogenesis is linked to a chronic low-grade inflammation and macrophage infiltration inside the adipose tissue, insulin resistance, cardiovascular disease and obesity. In the present paper we aimed to evaluate the role of metformin and vitamin D, alone or in combination, in modulating inflammation and autophagy in ADSCs during adipogenic commitment. ADSCs were cultured for 21 days in the presence of a specific adipogenic differentiation medium, together with metformin, or vitamin D, or both. We then analyzed the expression of FoxO1 and Heat Shock Proteins (HSP) and the secretion of proinflammatory cytokines IL-6 and TNF-α by ELISA. Autophagy was also assessed by specific Western blot analysis of ATG12, LC3B I, and LC3B II expression. Our results showed the ability of the conditioned media to modulate adipogenic differentiation, finely tuning the inflammatory response and autophagy. We observed a modulation in HSP mRNA levels, and a significant downregulation in cytokine secretion. Taken together, our findings suggest the possible application of these molecules in clinical practice to counteract uncontrolled lipogenesis and prevent obesity and obesity-related metabolic disorders.

Comparing Clinicopathologic Features and Surgical Treatment of Premenopausal Breast Cancer across Italy and China: Report from a Medical Exchange Program.

BACKGROUND: This study investigated the differences in clinicopathologic features and surgical treatment between an Italian and a Chinese cohort of premenopausal women with breast cancer, and highlighted the potential advantages of international medical exchange projects. METHODS: Premenopausal women who underwent surgical treatment between 2012 and 2016 at one Italian and one Chinese institution participating in a medical exchange program were compared. Factors associated with the probability to receive mastectomy were determined via logistic analysis. Changes in surgical management at the Chinese institution in the period 2018-2019, after the exchange program, were also evaluated. RESULTS: A total of 505 patients, 318 from Italy and 187 from China, were evaluated. The Chinese patients had more frequently advanced-stage tumours, large tumour size (30.9 vs. 18.1 mm, p < 0.01), invasive carcinoma (92.5 vs. 83.3%, p < 0.01), positive axillary lymph nodes (54.5 vs. 27.4%, p < 0.01), Her-2 positivity (36.4 vs. 22.0%, p < 0.01), and high proliferative index (55.1 vs. 30.2%, p < 0.01). Positive oestrogen receptor status and rates of triple-negative breast cancer did not differ (77.0 vs. 69.5%, p = 0.09 and 14.2 vs. 16%, p = 0.56, respectively). Mastectomy rates were higher among Chinese women (85 vs. 41%, p < 0.001), whereas use of sentinel node biopsy was more frequent among Italian women (77 vs. 33%, p < 0.001). Chinese women had more than 4-fold higher risk of receiving mastectomy. In the last 2 years, the rates of breast-conserving surgery and sentinel node biopsy at the Chinese institution increased from 15 to 23%, and from 33 to 42%, respectively. CONCLUSIONS: Tumour features and surgical strategies for premenopausal breast cancer may differ significantly between Italy and China. Since the international exchange program, patients from the Chinese institution have been offered more frequently less invasive surgery. International exchange programs can help in designing epidemiological studies which may be useful for strategies to improve breast cancer management and control.

Tuning Adipogenic Differentiation in ADSCs by Metformin and Vitamin D: Involvement of miRNAs.

Fat tissue represents an important source of adipose-derived stem cells (ADSCs), which can differentiate towards several phenotypes under certain stimuli. Definite molecules as vitamin D are able to influence stem cell fate, acting on the expression of specific genes. In addition, miRNAs are important modulating factors in obesity and numerous diseases. We previously identified specific conditioned media able to commit stem cells towards defined cellular phenotypes. In the present paper, we aimed at evaluating the role of metformin on ADSCs differentiation. In particular, ADSCs were cultured in a specific adipogenic conditioned medium (MD), in the presence of metformin, alone or in combination with vitamin D. Our results showed that the combination of the two compounds is able to counteract the appearance of an adipogenic phenotype, indicating a feedforward regulation on vitamin D metabolism by metformin, acting on CYP27B1 and CYP3A4. We then evaluated the role of specific epigenetic modulating genes and miRNAs in controlling stem cell adipogenesis. The combination of the two molecules was able to influence stem cell fate, by modulating the adipogenic phenotype, suggesting their possible application in clinical practice in counteracting uncontrolled lipogenesis and obesity-related diseases.

Unravelling Cellular Mechanisms of Stem Cell Senescence: An Aid from Natural Bioactive Molecules.

Cellular senescence plays a role in the onset of age-related pathologies and in the loss of tissue homeostasis. Natural compounds of food or plants exert an important antioxidant activity, counteracting the formation of harmful free radicals. In the presence of an intense stressing event, cells activate specific responses to counteract senescence or cell death. In the present paper, we aimed at evaluating the levels of expression of specific markers of senescence, in order to demonstrate that extracts from Myrtus Communis L. can prevent premature senescence in ADSCs exposed to oxidative stress. Cells were cultured in the presence of Myrtus extracts for 12-24 and 48 h and then incubated with H2O2 to induce senescence. We then evaluated the expression of senescence-related markers p16, p19, p21, p53, TERT, c-Myc, and the senescence-associated ß-Galactoidase activity. Our results showed that pre-treatment with Myrtus extracts protects cells from premature senescence, by regulating the cell cycle, and inducing the expression of TERT and c-Myc. These findings suggest a potential application of these natural compounds in the prevention and treatment of various diseases, counteracting premature senescence and preserving tissue functions.

Extracts from Myrtle Liqueur Processing Waste Modulate Stem Cells Pluripotency under Stressing Conditions.

Nutraceuticals present in food are molecules able to exert biological activity for the prevention and treatment of various diseases, in form of pharmaceutical preparations, such as capsules, cream, or pills. Myrtus communis L. is a spontaneous Mediterranean evergreen shrub, widely known for the liqueur obtained from its berries rich in phytochemicals such as tannins and flavonoids. In the present study, we aimed to evaluate the properties of myrtle byproducts, residual of the industrial liqueur processing, in Adipose-derived stem cells (ADSCs) induced at oxidative stress by in vitro H2O2 treatment. Cells were exposed for 12-24 and 48h at treatment with extracts and then senescence-induced. ROS production was then determined. The real-time PCR was performed to evaluate the expression of inflammatory cytokines and sirtuin-dependent epigenetic changes, as well the modifications in terms of stem cell pluripotency. The ß-galactosidase assay was conducted to analyze stem cell senescence after treatment. Our results show that industrial myrtle byproducts retain a high antioxidant and antisenescence activity, protecting cells from oxidative stress damages. The results obtained suggest that residues from myrtle liqueur production could be used as resource in formulation of food supplements or pharmaceutical preparations with antioxidant, antiaging, and anti-inflammatory activity.

Melatonin and Vitamin D Orchestrate Adipose Derived Stem Cell Fate by Modulating Epigenetic Regulatory Genes.

Melatonin, that regulates many physiological processes including circadian rhythms, is a molecule able to promote osteoblasts maturation in vitro and to prevent bone loss in vivo, while regulating also adipocytes metabolism. In this regard, we have previously shown that melatonin in combination with vitamin D, is able to counteract the appearance of an adipogenic phenotype in adipose derived stem cells (ADSCs), cultured in an adipogenic favoring condition. In the present study, we aimed at evaluating the specific phenotype elicited by melatonin and vitamin D based medium, considering also the involvement of epigenetic regulating genes. ADSCs were cultured in a specific adipogenic conditioned media, in the presence of melatonin alone or with vitamin D. The expression of specific osteogenic related genes was evaluated at different time points, together with the histone deacetylases epigenetic regulators, HDAC1 and Sirtuins (SIRT) 1 and 2. Our results show that melatonin and vitamin D are able to modulate ADSCs commitment towards osteogenic phenotype through the upregulation of HDAC1, SIRT 1 and 2, unfolding an epigenetic regulation in stem cell differentiation and opening novel strategies for future therapeutic balancing of stem cell fate toward adipogenic or osteogenic phenotype.

Red cell distribution width and mean platelet volume as predictors of anastomotic dehiscence in colorectal surgery A pilot multicenter case-match study.

AIM: One of the most serious complications in modern colorectal surgery is the occurrence of an anastomotic dehiscence. The aim of this study was to evaluate the role of preoperative red cell distribution width (RDW) and mean platelet volume (MPV) as predictors of anastomotic dehiscence in elective surgery for colorectal cancer. MATERIALS AND METHODS: Forty-two patients with a clinically manifested anastomotic dehiscence after oncological colorectal surgery, and 42 controls matched for age, sex, pathological stage and tumor localization were enrolled. Correlations between the preoperative RDW and MPV values and anastomotic dehiscence were investigated. RESULTS: Both the median RDW value (14.4 % vs 13.1%; p=0.007) and the median MPV value (8.0 fL vs 7.5 fL; p=0.037) were significantly higher in patients with anastomotic dehiscence than in those without. In multiple regression analysis only the RDW remained significantly associated with anastomotic dehiscence. CONCLUSIONS: The preoperative values of RDW may be useful in predicting anastomotic damage in elective oncological surgery. KEY WORDS: Anastomotic Dehiscence, MPV, RDW.

Dyspnoea, thoracic pain and fever in a young caucasian female: A case report.

INTRODUCTION: The diagnostic approach to patients with mediastinal pathology is not always simple and an improper diagnostic work-up can lead to significant diagnosis delay. PRESENTATION OF CASE: We report on the case of a young woman who was admitted to the Emergency Department complaining of thoracic pain, dyspnoea, fever and productive cough. The physical examination showed a painful swelling over the sternum's upper left margin, which had become evident 4 months earlier. A Computer Tomography showed the presence of a retrosternal oval lesion (5.5 x 4 cm) infiltrating the thoracic wall and showed the presence of discretely enlarged mediastinal lymph nodes in several mediastinal stations. DISCUSSION: The Multidisciplinary Team decided to perform an ultrasound-guided biopsy of the retrosternal mass that showed an inflammatory pattern, whereas microbiology tests proved negative. The lack of improvement with medical therapy (non steroideal anti-inflammatories and antibiotics) and the clinical suspicion of malignancy led us to perform a surgical biopsy of the mass that finally proved to be diagnostic for Hodgkin's lymphoma. CONCLUSIONS: Mediastinal masses with an aggressive behavior, should always be considered to be potentially malignant. Surgical biopsy, sometimes, can be the only way to correctly diagnose the pathological process, especially in the case of Hodgkin's lymphoma in which few diagnostic cells (Reed-Sternberg cells) are generally embedded in an abundant inflammatory background tissue.

Deciduoid mesothelioma of the thorax: A comprehensive review of the scientific literature.

OBJECTIVE: Deciduoid mesothelioma is a rare variant of malignant epithelioid mesothelioma. It often involves the peritoneum, but also thoracic cases have been reported. The aim of the present review is to describe the demographic, clinical, radiological, and pathological features of such a rare variant of thoracic mesothelioma, and the state of the art regarding the therapeutic approaches currently available. DATA SOURCE: English-language articles published from 1985 to June 2016, and related to thoracic deciduoid mesothelioma cases were retrieved using the Pubmed database. STUDY SELECTION: The search terms were "mesothelioma," "thoracic mesothelioma," "epithelial mesothelioma," "pleural mesothelioma," and "deciduoid mesothelioma." RESULTS: Forty-four cases included in 16 articles, published in the period under investigation, were analyzed in detail. CONCLUSIONS: The mean age of the patients was 63 years, and the male to female ratio 1.7:1. Approximately 58% had exposure to asbestos, and 73% had a smoking history; familiarity was rarely reported. The most common anatomical site of origin was the right pleura, and the most frequent clinical manifestations were chest pain, dyspnea, cough, and weight loss. Thoracic X-ray and computed tomography were the imaging techniques most employed for diagnosis and surgical planning. The pathological diagnosis was obtained by examination of surgical or biopsy specimens in most cases. The best treatment strategy of deciduoid mesothelioma is a matter of debate; nevertheless a multidisciplinary approach is currently the best option for the choice of the adequate therapeutic scheme.

Desmoplastic small round cell tumors of the pleura: a review of the clinical literature.

Desmoplastic small round cell tumor of the pleura is a rare malignancy, with only a few cases reported in the scientific literature. The aim of the present review is to discuss the demographic, pathological, clinical, and therapeutic features of this rare tumor. English-language articles published since 1989, when the first case of desmoplastic small round cell tumor of the pleura was described, were retrieved, and fifteen cases included in fourteen articles were revised. The mean age of the patients was 25.5 years, out of them 60% were males. Chest pain, pleural effusion, and dyspnea were the most common clinical manifestations, while chest roentgenogram and computed tomography were the imaging techniques most commonly used. Surgical biopsy was employed in 80% of the cases for diagnosis. A multidisciplinary approach consisting in a combination of surgery with chemotherapy and radiation therapy was adopted in most cases. Only two patients (13.3%) were alive at 3 years from diagnosis, reflecting the aggressiveness of the disease, and the poor outcomes of the treatments currently available. Desmoplastic small round cell tumors of the pleura are extremely aggressive and challenging to diagnose, because of their rarity and unspecific demographic, clinical, and radiological features. An in-depth knowledge of such features is necessary for the optimal management of patients with this rare malignancy.

Melatonin and Vitamin D Interfere with the Adipogenic Fate of Adipose-Derived Stem Cells.

Adipose-derived stem cells (ADSCs) represent one of the cellular populations resident in adipose tissue. They can be recruited under certain stimuli and committed to become preadipocytes, and then mature adipocytes. Controlling stem cell differentiation towards the adipogenic phenotype could have a great impact on future drug development aimed at counteracting fat depots. Stem cell commitment can be influenced by different molecules, such as melatonin, which we have previously shown to be an osteogenic inducer. Here, we aimed at evaluating the effects elicited by melatonin, even in the presence of vitamin D, on ADSC adipogenesis assessed in a specific medium. The transcription of specific adipogenesis orchestrating genes, such as aP2, peroxisome proliferator-activated receptor γ (PPAR-γ), and that of adipocyte-specific genes, including lipoprotein lipase (LPL) and acyl-CoA thioesterase 2 (ACOT2), was significantly inhibited in cells that had been treated in the presence of melatonin and vitamin D, alone or in combination. Protein content and lipid accumulation confirmed a reduction in adipogenesis in ADSCs that had been grown in adipogenic conditions, but in the presence of melatonin and/or vitamin D. Our findings indicate the role of melatonin and vitamin D in deciding stem cell fate, and disclose novel therapeutic approaches against fat depots.

Perineural infiltration as a prognostic factor in surgically treated gallbladder cancer A single center experience and literature review.

INTRODUCTION: Gallbladder cancer (GBC) is the most incident cancer of the biliary tract with only 5-13% of the sufferers surviving for five years. The aim of this study was to evaluate the prognostic role of perineural invasion (PNI) and its association with several clinicopathological variables in a cohort of surgically treated patients, and through a comprehensive review of the scientific literature. MATERIALS AND METHODS: Twenty-five consecutive patients submitted to curative surgery for GBC from 2008 through 2016 were enrolled. Demographic, clinical and pathological data were retrieved from medical files, and specimens were re-examined by two experienced pathologists. The Pubmed database was searched for articles reporting on perineural infiltration on gallbladder cancer. RESULTS: Perineural invasion was observed in 14 (56%) cases, and it was more frequent in higher pathological stages. A statistically significant association was found with high preoperative serum Ca 19-9 levels. Fourteen (56%) patients died during the follow-up; survival was lower in patients with perineural invasion in comparison to those without, but not statistically significant. Twelve English-language articles reporting on PNI were retrieved and discussed. CONCLUSIONS: Perineural invasion is associated with higher stage and poorer survival in surgically treated GBC patients. In patients with locally advanced GBC resection of the extrahepatic biliary duct and frozen section examination of the distal stump must be taken into consideration, especially in cases of tumor arising from the hepatic side of the gallbladder. In cases without residual disease but with pathological evidence of PNI, a careful follow-up is suggested to early detect recurrences. KEY WORDS: Adenocarcinoma, Cancer, Gallbladder, Perineural infiltration, Surgery.

Surgical outcomes in patients with hepatic synchronous and metachronous colorectal metastases.

AIM: To evaluate the impact of several clinical and pathological factors on the outcomes of surgery for hepatic colorectal cancer metastasis. METHODS: Eighty-four liver metastasectomies in 77 consecutive patients with 90 colorectal cancer hepatic metastases were performed in our institution from 2009 to 2014. Surgery was carried out in 75 cases, as two patients were not eligible for surgery. Among them 43 (Group A) were affected by synchronous, and 32 (Group B) by metachronous lesions. Furthermore, 9 reoperations were performed in patients with initially synchronous lesions. The follow-up after surgery included total body CT scan every 3 months for the first year, and every 6 months for 4 years thereafter. Blood level of CEA was determined every 3 months. RESULTS: The univariate analysis evidenced significantly more recurrences in patients with synchronous lesions (p=0.011), and higher grade, pN stage and CEA blood levels. In multivariate logistic regression analysis the statistically significant parameters found were: the pT stage (OR: 3.92, p = 0.039), the use of adjuvant chemotherapy for the colonic tumor (OR: 0.19, p = 0.025), and the adjuvant chemotherapy (OR: 4.11, p = 0.048). The global survival was 32 patients (41.5%), 17 with synchronous and 15 with metachronous lesions, and a significant difference in long-term survival between these two groups was found (p = 0.008). CONCLUSIONS: The most relevant prognostic factor in patients with hepatic colorectal cancer dissemination is the timing of metastasis; the metachronous lesions present better survival when surgically treated. KEY WORDS: Colorectal cancer, Liver, Metastasis, Surgery.

Surgical treatment of chronic multivascular mesenteric ischemia in a patient with antiphospholipid syndrome, abdominal aortic aneurism, and renal cancer: when planning overwhelms complexity.

Chronic mesenteric ischemia is a clinical condition caused by obstructive or occlusive disease of the mesenteric vessels, with potentially lethal consequences. We describe a case of open multiple revascularization in a patient affected by antiphospholipid syndrome and diffuse atherosclerosis, with an abdominal aortic aneurism, a contracted kidney, a renal cancer affecting the contralateral kidney, and as a consequence, a chronic renal failure and hypertension. We revascularized the celiac trunk, the superior and inferior mesenteric arteries, and the right renal artery using saphenous grafts; the aneurism was corrected, and the renal tumor was treated by radiofrequency ablation. Despite the invasiveness and complexity, the surgical strategy adopted allowed to save the patient's life, to treat the chronic mesenteric ischemia and the renal cancer, and to improve the chronic renal insufficiency and hypertension. KEY WORDS: Graft, Mesenteric ischemia, Occlusion, Revascularization.

Spigelian hernia A series of cases and literature review.

AIM: The aim of this case series is to describe the pathophysiological, clinical, and radiological features of Spigelian hernias, and to describe their modern surgical management. MATERIAL OF STUDY: We describe the clinical management of four cases of Spigelian hernias, and we discuss the main issues and controversies in the current scientific literature. RESULTS: In all the cases the clinical suspect arouses during the clinical examination of the patients, and it was confirmed through imaging techniques. All cases were treated surgically by an open approach; no intraoperative or postoperative complications occurred, and no recurrences were registered. DISCUSSION: Our experience and the literature review that we carried out confirm that Spigelian hernia is a rare condition, and its diagnosis should always be based on a thorough clinical examination and possibly a ultrasound or computed tomography, which are important for the study of the exact anatomical features of the defect, and for the planning of the adequate operative approach. The laparoscopic technique is effective but used in few reference centers, both for the rarity of the disease and the long learning curve, whereas the open surgical correction is the most common approach, with excellent results, especially in cases where urgent repair is needed. CONCLUSION: The surgical treatment of Spigelian hernias has excellent results when a thorough clinical - radiological evaluation is carried out, and the appropriate surgical approach is chosen. KEY WORDS: Abdominal wall, Spigelian hernia, Surgery.

Abdominal Wall Schwannoma.

Neurilemmomas-or schwannomas-are rare soft tissue tumours involving peripheral nerve sheaths, usually found in the head and neck regions. They can infrequently originate within the tissues of the abdominal wall. Here, we present a case of symptomatic schwannoma of the abdominal wall in a 62-year-old woman referred for abdominal pain in the right iliac fossa. On physical examination, a 5-7-cm oval-shaped area of consolidation with regular borders and elastic consistence was palpable. Ultrasound examination of the abdomen revealed a hypoechogenic mass measuring 80-33-42 mm; subsequently, a CT scan confirmed the presence of a well-circumscribed mass, with small calcifications inside. Radical excision of the lesion under general anaesthesia was performed, and the histological examination was consistent with the diagnosis of "ancient" schwannoma. The patient was discharged on the second postoperative day, and, at a clinical check 1 month postoperation, she reported no recurrence of abdominal pain and had an improved quality of life. Schwannomas have a good prognosis overall, with malignant degeneration being very rare. Local recurrence is plausible only if non-radical resection of the primitive tumour occurs. This is the second case ever reported, to our knowledge, of symptomatic schwannoma of the abdominal wall. We advocate surgical removal of the tumour when it presents as a cause of abdominal pain, ensuring that a radical excision is performed due to the possibility-though rare-of malignant transformation or recurrence. This offers the possibility of total regression of symptoms through surgical therapy.