Lidia Puzzo

Efficacy of adipose tissue-mesenchymal stem cell transplantation in rats with acetaminophen liver injury.

OBJECTIVE Acetaminophen intoxication is a leading cause of acute liver failure. Liver transplantation for acute liver failure is limited by the availability of donor organs. In this study, we aimed at identifying if the transplantation of adipose tissue-mesenchymal stem cells (ASCs) may exert therapeutic effects on acetaminophen-induced liver injury. METHODS ASCs were isolated from human subcutaneous tissue and were transfected with a green fluorescent protein (GFP). Sprague-Dawley rats were administrated 300mg/kg of acetaminophen intraperitoneally and were transplanted with ASCs or vehicle. After 24h from acetaminophen administration, rats were sacrificed. Hepatic levels of isoprostanes, 8-hydroxyguanosine (8-OHG), nitrites/nitrates and reduced glutathione (GSH) were determined as markers of oxidative stress; JNK phosphorylation and hepatic levels of inflammatory cytokines and regeneration factors were also assessed. RESULTS Transplantation of ASCs decreased AST, ALT and prothrombin time to the levels observed in control rats. Transplanted animals had normal plasma ammonia and did not display clinical encephalopathy. Liver sections of intoxicated rats treated with vehicle showed lobular necrosis and diffuse vacuolar degeneration; in rats transplanted with ASCs liver injury was almost absent. Transplantation of ASCs decreased liver isoprostanes, 8-OHG and nitrite-nitrates to the levels of control rats, while preserving GSH. Consistently, hepatic levels of TNF-α, MCP-1, IL-1β, ICAM-1 and phospho-JNK were markedly increased in rats treated with vehicle and were restored to the levels of controls in animals transplanted with ASCs. Furthermore, ASC transplantation increased liver expression of cyclin D1 and PCNA, two established hepatocyte regeneration factors, whereas ASCs were not able to metabolize acetaminophen in vitro. CONCLUSION In this study, we demonstrated that ASC transplantation is effective in treating acetaminophen liver injury by enhancing hepatocyte regeneration and inhibiting liver stress and inflammatory signaling.

Moro orange juice prevents fatty liver in mice

AIM To establish if the juice of Moro, an anthocyanin-rich orange, may improve liver damage in mice with diet-induced obesity. METHODS Eight-week-old mice were fed a high-fat diet (HFD) and were administrated water or Moro juice for 12 wk. Liver morphology, gene expression of lipid transcription factors, and metabolic enzymes were assessed. RESULTS Mice fed HFD displayed increased body weight, insulin resistance and dyslipidemia. Moro juice administration limited body weight gain, enhanced insulin sensitivity, and decreased serum triglycerides and total cholesterol. Mice fed HFD showed liver steatosis associated with ballooning. Dietary Moro juice markedly improved liver steatosis by inducing the expression of peroxisome proliferator-activated receptor-α and its target gene acylCoA-oxidase, a key enzyme of lipid oxidation. Consistently, Moro juice consumption suppressed the expression of liver X receptor-α and its target gene fatty acid synthase, and restored liver glycerol-3-phosphate acyltransferase 1 activity. CONCLUSION Moro juice counteracts liver steatogenesis in mice with diet-induced obesity and thus may represent a promising dietary option for the prevention of fatty liver.

miRNA profiling in vitreous humor, vitreal exosomes and serum from uveal melanoma patients: Pathological and diagnostic implications.

Uveal melanoma (UM) represents approximately 5–6% of all melanoma diagnoses and up to 50% of patients succumb to their disease. Although several methods are available, accurate diagnosis is not always easily feasible because of potential accidents (e.g., intraocular hemorrhage). Based on the assumption that the profile of circulating miRNAs is often altered in human cancers, we verified whether UM patients showed different vitreous humor (VH) or serum miRNA profiles with respect to healthy controls. By using TaqMan Low Density Arrays, we analyzed 754 miRNAs from VH, vitreal exosomes, and serum of 6 UM patients and 6 healthy donors: our data demonstrated that the UM VH profile was unique and only partially overlapping with that from serum of the same patients. Whereas, 90% of miRNAs were shared between VH and vitreal exosomes, and their alterations in UM were statistically overlapped with those of VH and vitreal exosomes, suggesting that VH alterations could result from exosomal dysregulation. We report 32 miRNAs differentially expressed in UM patients in at least 2 different types of samples analyzed. We validated these data on an independent cohort of 12 UM patients. Most alterations were common to VH and vitreal exosomes (e.g., upregulation of miR-21,-34 a,-146a). Interestingly, miR-146a was upregulated in the serum of UM patients, as well as in serum exosomes. Upregulation of miR-21 and miR-146a was also detected in formalin-fixed, paraffin-embedded UM, suggesting that VH or serum alterations in UM could be the consequence of disregulation arising from tumoral cells. Our findings suggest the possibility to detect in VH and serum of UM patients “diagnostic” miRNAs released by the affected eye: based on this, miR-146a could be considered a potential circulating marker of UM.

The Role of BMI1 as a Biomarker of Cancer Stem Cells in Head and Neck Cancer: A Review

Emerging studies show that BMI1 (B cell-specific Moloney murine leukemia virus integration site 1) has an important function as a biomarker of cancer stem cells (CSCs), i.e. cells with self-renewal characteristics, capable of tumor initiation, progression, invasion, metastasis, tumor recurrence and resistance to chemotherapy and radiotherapy. The failure of current anticancer therapies can be attributed to the relative ineffectiveness of drug and radiation treatments on CSCs, thereby preserving the full capacity of the cells to reproduce tumors. The development of new strategies is currently hindered by the lack of reliable markers for the identification of these CSCs. At present, they have been isolated from solid tumors at various locations using a variety of surface markers, including CD34, CD133, CD24, CD44, CD29 and CD31, in addition to the methods of isolation and cell culture via the Wnt, BMI1, Notch and Hedgehog pathways. The discovery of specific tumor targets for CSCs would constitute a big step in the research for the definitive therapy against cancer. More studies are being conducted that consider the role of CSCs in head and neck cancers with potential for an impact on clinical-surgical outcomes from the knowledge that is being gained. A promising intracellular marker of CSCs in head and neck cancer is the oncoprotein BMI1, with specific data about its prognostic value based on the specific location.

Nuclear BMI-1 expression in laryngeal carcinoma correlates with lymph node pathological status.

BackgroundThe main cause of treatment failure and death in laryngeal squamous cell carcinoma is metastasis to the regional lymph nodes. The current clinical staging criteria fail to differentiate patients with occult metastasis from patients without metastasis. Identifying molecular markers of the disease might improve our understanding of the molecular mechanisms underlying the pathogenesis and development of laryngeal carcinoma and may help improve clinical staging and treatment.MethodsSixty-four previously untreated patients who underwent surgical excision of laryngeal squamous cell carcinoma with neck dissection were included in this study. The expression of B cell-specific Moloney murine leukemia virus integration site 1 (BMI-1) was examined immunohistochemically on formalin-fixed paraffin-embedded primary tissue specimens.ResultsNuclear expression of BMI-1 (nBMI-1) was detected in 32 of the 64 tumors (50%), cytoplasmic expression of BMI-1 (cBMI-1) was detected in 22 (34.4%), and 10 tumors (15.6%) showed no BMI-1 immunoreactivity. High nBMI-1 expression levels (≥10) were detected in 28 of the 32 (87.5%) nBMI-1-positive patients. Multivariate analysis including age at diagnosis, grade, tumor location, TNM status, and nBMI-1 expression showed that a high nBMI-1 expression level was an independent prognostic factor for lymph node metastasis.ConclusionThe expression of BMI-1 in patients with laryngeal carcinoma seems to correlate with lymph node metastasis.

CD10 is expressed by mammary myofibroblastoma and spindle cell lipoma of soft tissue: an additional evidence of their histogenetic linking

Sir, CD10 is a cell-surface zinc metalloprotease expressed by a variety of normal and neoplastic cells. As far as the breast is concerned, CD10 stains normal and neoplastic myoephitelial cells, stromal cells of invasive ductal carcinoma and variably the stromal cell component of fibroadenomas and phylloides tumours [7]. Whilst it is a reliable myoepithelial cell marker, which can be helpful for diagnostic purposes in the differential diagnosis between benign sclerosing lesions and tubular carcinoma, its use is controversial in differentiating benign from borderline vs malignant phylloides tumours [7]. As stromal cell component in fibroademona and phylloides tumours is positive—even if with a variable extension—for CD10, we investigated its expression and distribution in mammary myofibroblastoma (MFB), a relatively unusual benign tumour, which arises from mammary stromal cells. To the best of our knowledge, there is no information about CD10 expression in mammary MFB. Accordingly, we performed an immunohistochemical study with anti-CD10 antibody (56C6 Novocastra, New Castle upon Tyne, UK; dilution 1:80; antigen retrieval using microwave oven) in a series of 12 cases of mammary MFBs (most of them already appeared in the form of original articles; see references [2–4]). A diffuse and strong cytoplasmic immunostaining to CD10 was observed in neoplastic spindle cells of most cases (11 out of 12 cases) (Fig. 1), whilst no reaction was found in 1 case. This novel finding suggests that CD10 should be included in the list of the immunohistochemical markers of MFB together with vimentin, desmin, α-smooth muscle actin, CD34, bcl-2, CD99 and estrogen, progesterone and androgen receptors [3, 4]. However, we would like to stress that diagnosis of MFB is mainly based on reliably morphological criteria [3, 4]. Immunohistochemistry is helpful, especially in its unusual morphological variants [2–4]. It is well-known that a close morphological resemblance exists between MFB and spindle cell lipoma (SCL), especially when the former contains a prominent fatty or myxoid component (so-called lipomatous or myxoid MFB) [2–4]. In addition, MFB and SCL share a basic common immunophenotype, their cells being positive to vimentin, CD34 and variably for bcl-2 and CD99 [3, 4]. The finding of identical karyotypic abnormalities in both MFB and SCL has provided additional evidence of the close relationship between these two tumours [1, 6]. The possibility that SCL may occur in the breast [3, 4], and vice versa, MFB do occur in soft tissues [5], renders their linking more intriguing. As the abovementioned overlapping clinical, morphological, immunohistochemical and cytogenetic features between MFB and SCL, we studied immunohistochemically the expression of CD10 even in a small series (10 cases) of soft tissue SCLs. It was noteworthy that the spindle cell component of all tumours showed a diffuse and strong cytoplasmic immunoreactivity to CD10 (Fig. 2). Virchows Arch (2007) 450:727–728 DOI 10.1007/s00428-007-0423-6

Two-as-one Monolateral Dual Kidney Transplantation

OBJECTIVES Dual kidney transplantation (DKT) of marginal kidneys could offer transplant candidates a very satisfactory kidney transplantation in terms of renal function. However, DKT might be considered a major surgical procedure and, in older recipients, has a potentially greater risk of surgical complications compared with single kidney transplantation. Because of these findings, some transplant centers have replaced the classic bilateral placement of 2 kidneys with the monolateral placement of both kidneys. METHODS In a group of 35 DKTs performed during a 5-year period, we applied a new technique of monolateral placement of DKT in 10 recipients. In these 10 patients, the arteries and veins of the 2 kidneys were joined through a running suture, and the joined kidneys were anastomosed into the external iliac vessels in the recipient. RESULTS The delayed graft function rate was 20%. No surgical complications developed in the entire series. One patient experienced late rejection with ureteral stricture. The graft and patient survival rate at a median follow-up of 30 months was 90%. CONCLUSIONS To reduce the surgical risk and morbidity rate, the monolateral placement of both kidneys seems the safest method to perform DKT. The joined monolateral DKT, by reducing the cold ischemia time and the surgical trauma, could represent a step forward in the delicate treatment of these patients.

Expression of BMI1 and p16 in laryngeal squamous cell carcinoma

The clinical evolution of laryngeal squamous cell carcinoma (SCC) is undetectable with the current staging criteria. To more completely understand the biology of laryngeal SCC, we assessed the expression of the proteins B‐cell–specific Moloney murine leukemia virus integration site 1 (BMI1) and p16.

Benign Prostatic Hyperplasia, Metabolic Syndrome and Non-Alcoholic Fatty Liver Disease: Is Metaflammation the Link?

The prevalence of prostatic inflammation (PI) is very frequent in patients affected by benign prostatic hyperplasia (BPH) and lower urinary tract symptoms (LUTS). To investigate the relationship between prostatic inflammation (PI) and the presence of MetS and non‐alcoholic fatty liver disease (NAFLD) in a cohort of patients affected by BPH/LUTS.

Cytomegalovirus and Clostridium difficile Ischemic Colitis in a Renal Transplant Recipient: A Lethal Complication of Anti-Rejection Therapy?

Intestinal ischemia is reported to be the most common gastrointestinal complication of renal transplantation and a potential cause of morbidity and mortality. The recent use of more potent immunosuppressive drug regimens has reduced the incidence of acute rejection, increasing the incidence of potentially fatal infectious complications, such as clinically important cytomegalovirus (CMV) infection. A 42-year-old kidney transplant recipient experienced on postoperative day 10 a dehiscence of the ureterovesical anastomosis, associated with a 7-cm longitudinal tear graft on the lower pole of the kidney and an ureteral ischemia. A graft biopsy demonstrated a mild acute rejection for which the patient received an unsuccessful administration of steroids, with progression of the rejection, so that 1 mg/kg/day antithymocyte globulin was administered. Two days later the patient presented with fever (39.5°C), diffuse abdominal pain with tenderness and bloody diarrhea, and diagnosis of CMV colitis was achieved; rectal samples were taken for histologic examination, and Clostridium difficile toxin was isolated. A subtotal colectomy with Hartmann’s procedure was performed, but the patient died 13 days later of a multiple organ failure. The risk of lethal CMV colitis is increased in patients being treated with anti-rejection therapy for severe acute rejection; the occurrence of simultaneous infection, such as pseudomembranous colitis, usually characterized by a favorable prognosis, increases the mortality rate in these patients.