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The aim of this study was to isolate and characterize rat Adipose Derived Mesenchymal Stem Cells (AD-MSCs) in order toevaluate their proliferative potential and their ability to differentiate in different cell types. AD-MSCs and Derived MesenchymalStem Cells (BM-MSCs) have the same characteristics in terms of plasticity. The advantage of adipose tissue is that it is an easier accessible source and it offers a large amount of MSCs by less invasive surgical tecniques. MSCs were obtainedfrom subcutaneous adipose tissue of Wistar rats. First of all microbiological controls were made to exclude the presenceof bacteria or fungi in the tissue. Adipose tissue was mechanically and enzimatically fragmented and stomal cell fraction was seeded in adherent culture flasks in DMEM 20% FBS. After 48h the medium was replaced. Cells were characterized byevaluating: 1)their ability to adhere to the plastic; 2) the clonogenic potential by Colony Forming Unit (CFU) assay; 3) theirability to differentiate in 3 mesodermal lineages (adipocytes, osteocytes and chondrocytes). AD-MSCs are able to differentiate in adipocytes, osteocytes and chondrocytes as confirmed by Oil Red’O staining, von Kossa staining and histologicalanalysis respectively. This first characterization is essential for the second part of our study in which we are planning to useAD-MSCs in vivo to restore renal function after an induced ischemic damage in experimental animals.

The term Diagnosis-related-Group (DRG) refers to a classification system used to assess hospital services with the aim of a better menagement of a health care costs and improving performance. The DRG system focuses on the utilization of resources, and is not concerned with the specific type of care provided to the patient. This system highlights any diseconomies and eventual critical aspectsof the hospital system. This article, starting from the history of heath care financing in Italy and pointing out the difficulty to define the "quality" of health care services, describes the variables used to evaluated correctly hospital performance based on the DRG system. These include Average Length of Stay, Average Daily Patient Load, Comparative Performance Index, and Case Mix Index.

Introduction The incidence of cancer compared for age groups is 3–4 times higher in transplant recipients than the general population. The increased risk is related to immunosuppressive therapy as well as the use of increasingly older donors and recipients. Although cardiovascular disease with a functioning transplant is the leading cause of death (47%), cancer mortality is significant especially among older patients. However, the most frequent posttransplantation cancers relate to hemolymphopoietic organs and skin, whereas the occurrence of solid tumors elsewhere is rare. Herein we have described a rare case of synchronous double malignancy of endocrine organs (thyroid-adrenal) in a young woman who underwent renal transplantation. Case Report A 37-year-old woman with end-stage renal disease for 18 years underwent transplantation when she was 30 years old with a 17-year-old standard cadaveric donor receiving immunosuppressive therapy with mycophenolate mofetil, cyclosporine, and steroids. Follow-up demonstrated good indices of renal function with negative tumor pathology at 79 months when, at an annual ultrasound monitoring, we found a lesion in the right lobe of the thyroid and left adrenal neoplasm of dubious interpretation. The cytology for the thyroid was highly suspicious of papillary carcinoma, whereas the histological examination after surgery diagnosed a thyroid multifocal papillary microcarcinoma (mpT1NxMx) and an oxyphil cell adrenocortical carcinoma (pT2, N0). Results Six months after total thyroidectomy with central lymphadenectomy and left kidney and adrenal gland removal the patient showed no evidence of recurrent lesions and stable graft function. Conclusions The rare occurrence of solid tumors after transplantation has no known etiopathogenetic relation. Despite the young age of the patient and the double neoplasm that could have produced an unfavorable outcome for the patient and the graft, careful follow-up for tumor pathologies and multidisciplinary management achieved an early diagnosis of both tumors with a surgical eradication without adjuvant therapy, preserving the life of the patient and the function of the graft.

We evaluated the incidence of and predisposing factors for an incisional hernia after kidney transplantation. Numerous techniques have been used to repair postoperative fascial dehiscences or simple incisional hernias, but no clear treatment exists for giant hernias. Our aim was to obtain (1) a safe procedure to repair a large abdominal defect and reinforce the surrounding, fragile zones and (2) a simple, rapid technique to reduce the operative time. Herein we have described the surgical repair of a giant incisional hernia using intraperitoneal Gore ePTFE dual-mesh plus (Gore-Tex; W. L. Gore, Flagstaff, Ariz, USA) in a 55-year-old man status-post renal transplantation. Total necrosis of distal graft ureter had caused a giant urinoma. The patient was reexplored on day 2 posttransplantation with a primary fascial approximation. Thirty days after transplantation we discovered a large incisional hernia and performed a repair. No drain was used. The patient continued immunosuppressive therapy (cyclosporine, mycophenolate mofetil, prednisolone) and was discharged on postoperative day 4 with no complications. An ultrasonographic follow-up at 1 year revealed the prosthesis to be correctely positioned. Incisional hernia is not rare after renal transplantation but the real incidence is unknown. Immunosuppressive therapy, prolonged pretransplantation dialysis, obesity, and diabetes are probably the major causes of incisional hernias in these patients. Surgical complications of renal transplantation surgery, such as wound hematoma, urinoma, and lymphocele, are the most important predisposing factors for an incisional hernia. The use of intraperitoneal ePTFE dual-mesh is feasible, safe, and easy to repair a large incisional hernia in a kidney transplant patient.

Early diagnosis of persistent hyperparathyroidism (HP) following kidney transplantation may prevent worsening of osteodystrophy and potential damage to the graft. We evaluated the utility of collagen pyridinoline (PYD) and deoxypyridinoline (DPD) urinary cross-links beyond the common HP markers to evaluate 70 selected stable recipients between 1997 and 2006 who were divided into 2 group depending on the immunosuppressive protocol. All patients showed elevated levels of urinary cross-links even though calcemia and phosphoremia values were normal. Their mean creatinine level was slightly increased. Data were assessed as mean values +/- SD. All variables underwent a correlation matrix analysis and a stepwise regression, with posttransplant intact parathyroid hormone (iPTH) as the dependent variable and other variables as regressors. A statistically significant correlation was observed between PYD and alkaline phosphatase (ALP; P = .0026, r = .41); PYD and DPD (P = .015, r = .34); pre- and posttransplant iPTH (P = .024, r = .31); and creatinine and ALP (P = .024, r = .31). Taking the groups separately, there were significant correlations between PYD and ALP (P = .0076, r = .42); PYD and DPD (P = .017, r = .38); ALP and posttransplant iPTH (P = .038, r = .33); osteocalcin (OC) and posttransplant iPTH (P = .048, r = .32); and pre- and posttransplant iPTH (P = .019, r = .37) among subjects in the first group, whereas subjects in the second group showed a correlation between posttransplant iPTH and age at transplantation (P = .032, r = .61). In conclusion, we showed that urinary cross-links may be helpful to reveal bone resorption in kidney recipients when usual bone metabolism parameters do not demonstrate hyperparathyroidism.

BACKGROUND: Aiming to deepen the understanding of the factors involved in the genesis of groin hernia, this study is focused on identifying the histological changes within the muscle fibers of the internal inguinal ring in patients having indirect inguinal hernia. METHODS: In eight patients with primary or recurrent bilateral indirect inguinal hernia who underwent a Stoppa open posterior inguinal hernia repair, a tissue specimen from the edge of the internal inguinal ring was biopsied and histologically examined. RESULTS: In all of the tissue samples, remarkable degenerative changes such as fibrohyaline degeneration of the muscle fibers, vascular congestion, and phlogistic infiltration through lymphohistiocytary elements was constantly detected. Also, in the patients with recurrent hernia, the key characteristic of the muscular change was that of fibrohyaline and, occasionally, myxoid degeneration of the myocytes. Nerve endings were frequently detected within the muscular structures of the internal inguinal ring. CONCLUSION: The degenerative fibrohyaline alteration, as well as the evidence of phlogistic elements within the examined structures, could represent a reason for a contractile incompetence of the internal inguinal ring. Consequently, the described findings lead the authors to depict this inflammatory degenerative structural weakness of the internal inguinal ring as a possible culprit of indirect inguinal hernia formation.

Secondary hyperparathyroidism (HP) presenting with hypocalcemia and subsequent increased parathormone (PTH), is mainly identified in patients with chronic renal failure, which has been associated with variable degrees of bone marrow fibrosis. For suitable patients with end-stage renal disease (ESRD), kidney transplantation is recognized as the therapy of choice, being superior to dialysis in terms of quality of life and long-term mortality risk; in this regard interesting data show that increased time on dialysis prior to kidney transplantation is associated with decreased graft and patient survival. In our opinion an important and until now underestimated determinant of graft survival is the proper activity of bone marrow because of the emerging role of hematopoietic stem cells (HSC) in repair of ischemia/reperfusion (IR) damage. We postulate that in ESRD patients, who usually undergo long dialytic treatment, a myelofibrosis caused by an overt secondary HP could drastically decrease the HSC potential for IR damage repair after kidney transplant; this could irremediably lead to a delay in graft function with all related complicances. If the curative role of bone marrow-derived stem cells was confirmed by more data obtained in experimental animal models, it could be possible to try a cellular-based therapeutic approach in the management of ESRD patients which are in waiting list for a kidney transplant.

Human intestinal microbiota create a complex polymicrobial ecology characterized by high population density, wide diversity, and complexity of interactions. Any imbalance of this complex intestinal microbiome, both qualitative and quantitative, might have serious health consequences including an increase in the number and/or alteration in the type of bacteria in the upper gastrointestinal tract, which is referred to as small intestinal bacterial overgrowth (SIBO) syndrome. SIBO is frequently found in persons fulfilling criteria for irritable bowel syndrome (IBS), and the large overlapping of symptoms of these two pathological conditions led some authors to believe that IBS is secondary to SIBO. Interestingly, SIBO is also found in about 25% of patients with Crohn's disease. Emerging data show that specific components of gut microbiota, particularly segmented filamentous bacteria (SFB), activate intestinal immunocompetent cells, for example, Th17 cells which have a potential role in pathogenesis of inflammatory intestinal diseases. On the basis of the aforementioned data we postulate that a previously unidentified specific form of SIBO, involving in particular the aberrant expansion of SFB in the gut, could play a role in the onset of chronic intestinal inflammatory diseases through persistent activation of Th17 cells. From this point of view, a successful therapeutic approach to inflammatory intestinal disease patients could be the administration of specific antibiotics directed against SFB to restore the physiological levels of these bacteria in the gut. Furthermore, it could be very useful to identify appropriate laboratory methodologies to monitor the level of SFB in the gut with the aim of preventing their potentially dangerous increase in numbers.

This report describes the use of the intraoperative parathyroid hormone (ioPTH) assay during parathyroidectomy in waiting list and transplanted patients. ioPTH levels were determined in 40 patients on the waiting list for kidney transplantation with secondary hyperparathyroidism who underwent subtotal parathyroidectomy and 9 transplanted patients with tertiary hyperparathyroidism who underwent removal of hyperplasic glands. Rapid PTH levels decreased significantly at each time period; the percentage decrease in rapid PTH levels was 61.3% among patients with IPT II and 70.2% in patients with IPT III at 10 minutes and 86.5% in patients with IPT II and 91% in patients with IPT III at 15 minutes after excision of hypersecreting parathyroid tissue. A decrease of 50% or more from baseline PTH levels at 10 minutes and/or a decrease of 85% or more at 15 minutes predicted successful removal of abnormal parathyroid glands. The application of this technique during subtotal parathyroidectomy has proved useful for correct excision of parathyroid glands among waiting list patients with IPT II, while in kidney transplant patients with IPT III it allowed removal of only the pathological glands with a limited surgical approach.

With the high occurrence of cardiovascular disease and increasing numbers of patients requiring vascular access, there is a significant need for small-diameter (

Tension-free muscle closure is essential in kidney transplantation, both in adult and pediatric patients. Tight muscle closure may lead to renal allograft compartment syndrome either due to compression of the renal parenchyma or due to kinking of the renal vessels. It may also cause kinking of the transplant kidney ureter, wound dehiscence and incisional hernia. Many techniques have been proposed in an attempt to achieve tension-free closure. There is a wrong belief among surgeons that using prosthetic mesh may increase the incidence of infective complications in these immunosuppressed patients. Also, there is fear that one is not able to monitor the renal graft by ultrasound and perform biopsy in the presence of a mesh. Other alternative techniques to mesh closure include subcutaneous placement and intraperitonealization of the kidney transplant. These techniques however, are valuable when mesh closure is unfavorable or contraindicated as in case of a potential source of infection, like a stoma. Abdominal wall fasciotomy can be adjunctive to the various techniques of muscle closure.Abdominal mesh closure, Post transplant incisional hernia, Renal transplantation, Renal Allograft Compartment syndrome (RACS).

The aim of this study was to isolate and characterize rat adipose Derived Mesenchymal Stem Cells (AD-MSCs) in order to evaluate their proliferative potential and their ability to different cell types. AD-MSCs and Derived Mesenchymal Stem Cells (BM-MSCs) have the same characteristic in terms of plasticity. The advantage of adipose tissue is that it is an easier accessible source and it offers a large amount of MSCs by less invasive surgical tecniques. MSCs were obtained from subcutaneous adipose tissue of Wistar rats. first of all microbiological controls were made to exclude the presence of bacteria of fungi in then tissue. Adipose tissue was mechanically and enzimatically fragmented and stromal cell fraction was seeded in adherent culture flasks in DMEM 20% FBS. After 48 h the medium was replaced. Cells were characterized by evaluating:1)their ability tho adhere to the plastic; 2) the clonogenic potential by Colony Forming Unit (CFU) assay, 3) their ability to differentiate in 3 mesodermal lineages (adipocytes, osteocytes and chondrocytes). AD-MSCs are able to differentiate in adipocytes, osteocytes and chondrocytes as confirmed by oil red'O staining, von Kossa staining and histological analuysis respectively. This first characterization is essential for the second part of our study in which we are planning to use AD-MSCs in vivo to restore renal function after an induced ischemic damage in experimental animals.

Background: Patients who underwent primary inguinal hernia repair still report a high rate of postoperative pain after operation due to effect of mesh fixation by suture. An alternative is the use of human fibrin glue. We compared the two techniques. Methods: 468 patients randomly underwent primary inguinal hernia Lichtenstein repair fixing the mesh by suture or by human fibrin glue (HFG); in both cases themesh was fixed to the posterior wall of the inguinal canal and to the inguinal legament. results: No significant differences were recorded between the two groups in terms of complications, while the sutureless technique reduces the operative time and the postoperative pain. Conclusions: A widespread technique for the treatment of inguinal hernia is the application of a mesh using lichtenstein procedure. The prosthesis can be fixed by traditional suture or using a new method of sutureless fixation with adesive materials that shows an excellent local tolerability and lack of adverse effects and contraindications.

Article 2 of the European Directive 2011/18/EC defines a genetically modified organism (GMO) a san organism in which the genetic material has been altered in a way that does not occuirr naturally by inserting DNA fragments from one organism's gene into the chromosomes of another, changing its genetic expression. In the field of animal and plants biotechnology there are different application areas of genetic engineering such as improving the quantity and quality of argicultural production, pharmaceutical production in more sustainable ways or applications in environmental field.

Exogenous insulin is, at the moment, the therapy of choice of diabetes, but does not allow tight regulation of glucose leading to long-term complications. Recently, pancreatic islet transplantation to reconstitute insulin-producing cells, has emerged as an alternative promising therapeutic approach. Unfortunately, the number of donor islets is too low compared with the high number of patients needing a transplantation leading to a search for renewable sources of high-quality -cells. This review, summarizes more recent promising approaches to the generation of new -cells from embryonic stem cells for transdifferentiation of adult cells, particularly a critical examination of the seminal work by Lumelsky et al.

Marginal donors represent a poorly utilized source of organs for transplantation despite their availability. The key is to reduce the ischemic damage in the effort to improve organ quality. This study investigated the histologic effects after in situ perfusion of preservation with a two-layer method compared with the classic University of Wisconsin preservation in term of tissue integrity and number of viable exocrine cells in the rat pancreas both after exsanguination and at 8 weeks of cryopreservation. Pancreata harvested from 60 rats were collected using 3 methods: two-layer method following University of Wisconsin perfusion; exsanguination; and classic University of Wisconsin perfusion/storage. In addition to histologic analysis of collected pancreata, we analyzed the number of CK19(+) cells and their viability using chi-square tests with values P < .05 considered to be significant. Rat pancreas histology showed as University of Wisconsin in situ perfusion and preservation by the two-layer method to be more effective to maintain the morphologic integrity of both exocrine and endocrine tissues. There were a larger number of CK19(+) cells with good viability. Moreover, the effects of oxygenation were visible in pancreas biopsies preserved after exsanguination. In situ University of Wisconsin perfusion and preservation for 240 minutes with the two-layer method yielded greater numbers and viability of CK19(+) cells even after 8 weeks of cryopreservation.

Colonic leiomyoma is a mesenchymal tumor that arises from the muscularis mucosae or muscularis propria and is composed of well-differentiated smooth muscle cells with no atypia. It is often incidentally found since its growth affects the submucosal layer and the lesion is covered with normal epithelium. Endoscopic ultrasonography is recommended to define the grade of infiltration of the tumor and eventually lymph node involvement. Histological examination is critical to establish the nature of the tumor and its behaviour. In the case of a voluminous tumor surgical treatment is needed. we report case of a patient that underwent colonoscopy showing the presence of a neoformation at 70 cm from ileocecal valve occupying half lumen of transverse colon. A surgical resection was performed and histological analysis confirmed the presence of a leiomyoma.

Tertiary hyperparathyroidism is intended as a state of excessive autonomous excretion of parathyroid hormone after longstanding secondary hyperparathyroidism. It is a condition of a severe hyperparathyroidism that persists in the renal recipient despite a successful transplantation. The high levels of parathyroid hormone lead to a worsening of graft function accompanied by systemic symptoms. This article reviews the main aspects related to tertiary hyperparathyroidism from mechanism of pathogenesis to prevention and treatment, suggesting some indications about the best management of patients on a waiting list for kidney transplant.

Background: Some uremic patients suffering from diabetes, atherosclerosis or simple by absent or exhausted peripheral vascular bed require bigger effort in creating arteriovenous fistula. Although it is possible to use prosthetic graft for difficult cases, the use of native vessels falls always in the respect of patient’s quality of life. A help for these cases comes from the use of a microsurgical technique for creating vascular access. Patient and Methods: For this study we evaluated the 72 patients underwent microsurgical radiocephalic fistula comparing them to a homogeneous group of 72 patients underwent traditional surgery. The patients were followed up at 1, 2 months and 1 year. Results: From comparison between the two groups our findings suggested that microsurgical radiocephalic arteriovenous fistula is more effective than traditional surgery in term of functional patency with appropriate dialysis. Conclusion: The use of microsurgical technique in our experience provides an improvement for the success of distal radiocephalic arteriovenous fistula in term of primary patency in those cases with a poor prognosis according to quality of distal native vessels.

Human intestinalmicrobiotais characterizedby highpopulationdensity, wide diversity and complexity of interactions. Dysbiosis, both qualitative and quantitative, might have serious health consequences including an increase in the number and/or alteration of bacteria colonizing the gastrointestinal tract, which could be referred as gut microflora imbalance (GMI). GMI is frequently found in patients suffering from irritable bowel syndrome; this is the reason why some authors believe that IBS is secondary to GMI. Interestingly, GMI is also found in about 25% of Crohn’s disease patients. Stimulation of inflammation by the ‘dangerous bacteria’, through specific activation of the intestinal immune system, seems to be the widely accepted pathogenetic hypothesis to explain such a correlation; on the contrary, some components of the intestinal microflora could also exert a protective role through immunomodulation. Stimulation of inflammation is also the main mechanism by which the gut microflora could increase the cancer risk; another alternative, but not mutually exclusive, pathogenetic hypothesis involves the regulation of proliferation/differentiation of intestinal stem cells. In this review, some interesting data regarding biomolecular alterations associated with intestinal bowel diseases will be described. We will discuss more recent findings about the relationship between GMI and intestinal chronic inflammation, pointing out the oncogenic risk related to alteration of microbiota composition. Finally, we generate an intriguing hypothesis regarding a network between gut microflora and intestinal stem cells. ! 2012 Wolters Kluwer Health | Lippincott Williams & Wilkins Reviews in Medical Microbiology 2012, 23:52‐57

Ras proteins are small GTPase functioning as molecular switches that, in response to particular extracellular signalling, as growth factors, activate a diverse array of intracellular effector cascades regulating cell proliferation, differentiation and apoptosis. Human tumours frequently express Ras proteins (Ha-, Ki-, N-Ras) activated by point mutations which contribute to malignant phenotype, including invasiveness and angiogenesis. Despite the common signalling pathways leading to similar cellular responses, studies clearly demonstrate unique roles of the Ras family members in normal and pathological conditions and the lack of functional redundancy seems to be explainable, at least in part, by the ability of Ras isoforms to localize in different microdomains to plasma membrane and intracellular organelles. This different intracellular compartmentalization could help Ras isoforms to contact different downstream effectors finally leading to different biological outcomes. Interestingly, it has also been shown that Ha- and Ki-Ras exert an opposite role in regulating intracellular redox status. In this regard we suggest that H-Ras specific induction of ROS (reactive oxygen species) production could be one of the main determinants of the invasive phenotype which characterize cancer cells harbouring H-Ras mutations. In our hypothesis then, while K-Ras (not able to promote oxidative stress) could mainly contribute to cancer progression and invasiveness through activation of MAPK and PI3K, H-Ras-mediated oxidative stress could play a unique role in modulation of intercellular contacts leading to a loss of cell adhesion and eventually also to a metastatic spread.

The gut of a healthy adult harbours a myriad of different microbial species. It is estimated that approximately 10 14 are present in total bacterial colony forming units (CFU). Each colony colonizes a specific intestinal tract. In healthy adult, the main control of intestinal bacterial colonization occurs through gastric acidity but also other factors can influence the intestinal microenvironment such as pH, temperature, competition among different bacterial strains, peristalsis, drugs, radiotherapy and much more. Impaired microbial homeostasis leads to an alteration of the permeability of tissue, together with the activation of the intestinal immune system MALT (mucosal associated lymphoid tissue). In this regard we discuss the increasing experimental evidences of the role of commensal microbiota in the activation of specific intestinal immunocompetent cells. The aforementioned micro-environmental changes provide the substrate for the etiopathogenetic outbreak of numerous pathologies of gastro-intestinal tract, such as intestinal chronic inflammation (Crohn's disease and Ulcerative Colitis), together with a miscellany of extra intestinal disorders. This article is an overview of the latest scientific findings about the close causal relationship between intestinal microbial flora and inflammatory bowel diseases or other extra-intestinal diseases; it is also mentioned the possible relationship between mycobacteria and Chron's disease. Finally we analyse the beneficial role of probiotics.

Background: Calciphylaxis is a potentially fatal complication of persistent secondary hyperparathyroidism; its cause is still not clear. Unfortunately there is no close relation in severity of clinical picture, serological and pathological alteration. For this reason the prognosis is difficult to establish. Administration of sodium thiosulphate may reduce the precipitation of calcium crystals and improve the general clinical conditions before surgical parathyroidectomy, which seems the only therapeutic approach able to reduce the mortality risk in these patients. Methods and Results: A 60 year old female patient suffering from End Renal Stage Disease, on haemodialysis from 2001 due to the onset of haemolytic uremic syndrome, underwent a kidney transplant in April 2008. After transplantation there was a recurrence of the haemolytic uremic syndrome, with temporary worsening of the graft. Six months later there was a definite loss of graft and return to dialysis treatment. On April 2010 a severe systemic calciphylaxis related to secondary hyperparathyroidism was diagnosed. The patient underwent parathyroidectomy but, because of the unimproved clinical picture, treatment with sodium thiosulphate was initiated. There was only improvement in cutaneous lesions. The worsening general clinical condition of the patient caused death due to general septic complications. Conclusions: The coexistence of haemolytic uremic syndrome and secondary hyperpathyroidism makes the prognosis poor and, in this case, therapy, which counteracts calcium crystals precipitation, has no effect. Preventive parathy­roidectomy can be considered as the only possible treatment.

Purpose Assessment of the effectiveness of Brachial Plexus Block (BPB) via axillary approach compared to regional anesthesia for arteriovenous fistula surgery in patients affected by end-stage renal disease. Methods We compared forty patients randomly divided into two groups. Group A underwent BPB procedure with 15 mL ropivacaine 1% and 10 mL of saline (0.9% NaCl) via axillary approach. Group B received local anesthesia with lidocaine 2%. The forearm blood vessels were assessed by Doppler ultrasonography before and after the intervention. Results BPB performed on Group A was associated with a considerable venous dilation and a significant decrease (48.7%, PConclusions The axillary-approached BPB was more advantageous than local anesthesia. Its effectiveness was because of venous dilation and the decrease in the PI, consequent to the reduction in peripheral resistances and the increase in local blood flow, thus offering an ideal background for fistula creation and short-term patency.

Purpose Pancreata from non-heart beating donors could represent an unlimited source of islets if their cell viability can be efficiently preserved during the time necessary to process the organs by the use of a better solution of preservation compared to the classic University of Wisconsin solution. The aim of this study was to determine whether it is possible to obtain functioning “alive islets” from non-heart-beating donors by comparing, on a porcine model, the classic “UW ice-store” method with a two-layer cold storage method (TLM) using oxygenated Perfluorocarbons (PFC) and UW. Methods Whole pancreata were harvested from 20 NHBDs female pigs with similar characteristics and preserved for 4 h in UW solution (n=10) or TLM (UW/PFC) solution (n=10). The isolated islets were then evaluated for number, viability, purity, and insulin secretion, also estimated after 8 weeks of cryopreservation. Results The total number of islets obtained from isolation, and their function assayed by the insulin stimulation index, before and after cryopreservation, showed a higher value in the TLM group. No significative differences in terms of purity and viability before and after cryopreservation were found when comparing the two groups. Conclusions TLM solution for NHBDs porcine pancreata with cold ischemia time lower than 4 h offers significant advantages over UW solution storage, thereby increasing the isolation yield and isolation success rate of the pancreatic porcine islets.

Ischemia is the most important factor that affects organ survival during harvesting. The two-layer method (TLM) is one of several cold storage solutions that seeks to preserve organs and cells avoiding in vivo and in vitro ischemia. We compared the retrieval of beta-like elements from exocrine pancreatic cells using TLM versus University of Wisconsin (UW) solutions. For this purpose pancreata laparoscopically harvested from 20 female pigs were preserved in UW solution or TLM before digestion. The resulting exocrine cells were divided into 2 groups: the first was cultured in a designed medium to allow differentiation into beta-like cells and the second was cryopreserved before the differentiation process at -196 °C for 8 weeks before culture in the same medium. The results revealed that TLM was better than UW as a preservation solution in terms of beta-cell viability and insulin secretion. We suggest that the use of TLM solution allows one to obtain less damaged cells for research purposes.

Struma ovarii is a rare form of ovarian neoplasm, entirely or predominantly composed of thyroid tissue. This tumour generally has a benign biological behavior, although some cases of malignant transformation have been reported (5-10%). The tumour usually presents as an asymptomatic mature mass that may measure as much as 10 cm in diameter. The presence of ascites is possible (15-20%), and in a few cases the association of ascites and hydrothorax has been recorded (Pseudo-Meigs Syndrome); a close relationship between ascites and high levels of Ca-125, that may wrongly induce to a diagnosis of ovarian carcinoma is present. Other possible clinical consequences are compressive symptoms on adjacent structures and symptoms of estrogenic-progestinal hyperproduction such as menometrorrhagia, dysmenorrhoea, infertility, and skin rash.Here we report the case of a young woman presenting, at her admission, a voluminous abdomino-pelvic mass (max. diam. 30 cm), histologically represented by a mature teratoma prevalently composed of thyroid tissue. Eur. J.Oncol., 16 (3), 000-000, 2011

A simple technique for implanting a long-term jugular catheter in piglets under general anesthesia is described. We report our experience in 10 young female pigs with a body weight of 20-30 Kg. The surgical procedure involves implantation of a jugular central venous catheter (11Fr polyurethane) tunneled in the subcutaneous fat layer of the neck. This procedure may be performed in about 15 minutes. The maintenance of the catheter is described which allows several daily blood samples to be taken. This procedure reduces both the stress in piglets and the chance of catheter dislodgement due to the animals scratching or rubbing. Blood sampling can be easily performed with a low incidence of infection or thrombosis

We sought to develop a protocol to isolate and culture porcine Wirsung duct cells in order to determine their potency to differentiate into insulin-expressing beta-like cells. The porcine Wirsung duct isolated by a surgical microdissection was digested with collagenase P and trypsin to dissociate ductal cells. These elements were cultured in serum-free supplemented media: for 2 weeks. Thereafter the cells were exposed to varying concentrations of glucose (0, 5.6, 17.8, and 25 mmol/L) to induce a beta-like phenotype, as identified by immunohistochemical staining. Cell growth proceeded slowly for the first 2 weeks of culture. After glucose induction for 2 weeks, they formed pancreatic islet-like structures. These cells were stained for the pancreatic ductal cell marker cytokeratin-19 (CK-19) and the pancreatic endocrine markers insulin and glucagon. After the second week, 90% of cells were positive for CK-19. Up to 20.1% of the cells in pancreatic 3-dimensional structures induced by 17.8 mmol/L glucose were positive for insulin, and

The aim of this work was to demonstrate a greater number of viable cells using a micro-surgical in-situ perfusion to collect rat pancreata compared with the pancreas after exsanguination. We used 3 groups of 20 rats. Perfusion was performed by selective cannulation of the left common iliac artery with administration of UW solution at 4 degrees C. Collected pancreata were digested and cells separated by Ficoll gradient were placed in culture to permit adhesion to dishes. Cells were characterized and tested for viability. We observed a gain of about 14% in the number of viable cells compared with those obtained after exsanguination (P < .001 by chi-square).

Introduzione:Fournier ha descritto per la prima volta una fascite necrotizzante dei genitali e/o del perineo che può evolvere rapidamente in shock settico con mortalità fino al 67%1. Caso Clinico: Pz. maschio 50 a.a. con turbe psichiche e diabete mellito II sottoposto ad emorroidectomia. A 24 ore dolore distrettuale, iperpiressia, edema dello scroto, crepitio inguinale sottocutaneo bilaterale. Veniva effettuato drenaggio chirurgico urgente della regione inguinale bilateralmente e recentazione scrotale; nel post-operatorio NPT e terapia antibiotica e.v.(ertapenem sodico 1 gr/die e linezolidd 600 mg x 2/die). Doppia medicazione/die e curettage dei reliquati necrotici seguiti da lavaggi con betadine, iruxol uso topico e zaffaggio con garze; 10 sedute di OTI2 a giorni alterni. Chiuse per prima intenzione le ferite inguinali, l’exeresi graduale delle sinechie tra derma e residui di dartos nonché l’aspirazione dell’idrocele facevano guadagnare stoffa cutanea e spazio per il packing. Quest’ultima metodica ripetuta per tre volte con un volume sempre maggiore di garze stipate consentiva l’espansione cutanea in due settimane con ricostruzione completa della borsa scrotale senza necessità di chirurgia plastica. Discussione/Conclusione: La tumefazione dello scroto e la presenza di crepitio del sottocutaneo inguinale ha consentito una diagnosi precoce con trattamento intensivo multidisciplinare immediato finalizzato a: a) risoluzione lo shock settico; b) eradicazione il focolaio infettivo inguino-scrotale; c) demarcazione della necrosi. Per la ricostruzione dello scroto, quando possibile, proponiamo questo metodo di espansione cutanea purchè si elidano le sinechie tra derma e residui di dartos. Nel nostro paziente nonostante si era avuta una perdita di circa due terzi della cute scrotale grazie alla mobilità testicolare ed al packing ripetuto è stato possibile ricostruire la borsa scrotale. La gangrena di Fournier è in prima istanza una emergenza chirurgica gravata da alta mortalità che necessita di trattamento dell’infezione distrettuale e dello shock settico. La ricostruzione scrotale, quando possibile, deve mirare alla restituzione della naturale elasticità dei residui cutanei accostabili sotto tensione con packing espansivo progressivo. In caso contrario deve intervenire la chirurgia plastica.

INTRODUCTION AND AIMS: One of most common complication following kidney transplantation is reappearance of hyperparathyroidism (HP) some months after normal serum levels. An early diagnosis represents the best prophylaxis measures to prevent worsening of osteodystrophy and potential damage to the graft and vascular tissue.The aim to our study was to evaluate, beyond the common HP markers, if the urinary cross-links of the collagen PYD (pyridinoline) and DPD (Deoxypyridinoline) are useful markers able to show increasing levels of parathormone. METHODS: We evaluated 50 stable kidney transplanted patients divided into 2 group (A: 38 pts and B: 12 pts) dependent on the type of their immunosuppressive therapy (Cyclosporine A and mycophenolate mophetil with or without corticosteroids). RESULTS: All patients showed elevated levels of urinary cross-links even though calcemia and phosphoremia values were normal and creatinine level slightly increased.( Table 1) A statistically significant correlation was found between PYD and ALP (alkaline phosfathase) (p=0.0026), PYD and DPD (p=0.015), pre- and post- kidney transplantation iPTH (intact parathyroid hormone) (p=0.024), and creatinine and ALP (p=0.024). Post kidney transplantation iPTH values were moderately elevated. No significant correlation was found between urinary cross-links and the other parameters. Taking the groups separately, while a significant correlation between PYD and ALP (p=0.0076), PYD and DPD (p=0.017), ALP and post transplant iPTH (p=0.038), OC (osteocalcin) and post transplant iPTH (p=0.048), and pre- and post-transplant iPTH (p=0.019) was found in subjects belonging to group A, subjects of group B showed a correlation between post-transplant iPTH and age at transplantation (p=0.032).(Table 2.) CONCLUSIONS: Our results showed that urinary cross-links could play a role in revealing bone reabsorption in kidney transplanted patients when usual bone metabolism parameters do not demonstrate a state of HP.

Objectives: We evaluated the incidence of incisional hernia after kidney transplantation, predisposing factors and the results of surgical repair of a giant incisional hernia by intraperitoneal PTFE dual mesh. Patients and methods: We report our experience about an incisional hernia in a 55 years male after renal transplantation treated with an intraperitoneal dual mesh prosthesis. The cause of incisional hernia was probably due to an urinoma for a total necrosis of distal graft ureter that needed to reopen in 2sd day post transplant with primary fascial approximation. Thirty days after transplantation we discovered a large incisional hernia that rapidly became monstrous with migration of omentum and small bowel in the sac. The giant hernia was repaired with a tension free technique by intraperitoneal positioning of a PTFE dual-mesh prosthesis, laterally anchored to inguinal ligament with 2-0 interrupted polipropilene stitches. Medially the prosthesis was anchored to peritoneal plane by interrupted transparietal stitches with an overlap of 5 cm. After a 2-0 polipropilene 180°double running suture was performed around the hernial border with an overlap of 2 cm. No drain was used. The patients continued his immunosuppressive therapy (CyA, MMF, prednisolone). Results: The patients was discharged in 4th day after operation with no complications. After six months ultrasonographic follow up, the prosthesis was still positioned between the inguinal ligament and posterior abdominal wall Conclusions: Incisional hernia is not rare after renal transplantation but the incidence is unknow. Immunosuppression based on corticosteroids and a higher prevalence of co-morbidity, such as obesity, diabetes, chronic pulmonary disease and others, probably are the causes of a major incidence of incisional hernia in these patients. Surgical complications of renal transplantation surgery, such as wound hematoma, urinoma, lymphocele are the most important predisposing factors for incisional hernia. Finally in our country, transplanted patients have higher risk of incisional hernia due to a long period of dialysis while awaiting transplantation. In these patients the use of intraperitoneal PTFE dual-mesh is feasible, safe, and easy.

INTRODUZIONE Il reintervento per patologia a carico della ghiandola tiroidea è raro, ma risulta spesso indicato nelle ricomparsa di patologia nodulare dopo interventi parziali. La ricorrenza della patologia multinodulare si aggira infatti intorno al 12% di tutti gli interventi sulla tiroide. Questi interventi di totalizzazione possono talvolta richiedere la collaborazione di equipe chirurgiche con diverse specialità in successivi steps temporali. MATERIALI E METODI Il caso clinico riguarda una donna di 72 anni sottoposta 40 anni prima ad intervento di tiroidectomia parziale che riferisce la comparsa di voluminosa tumefazione in sede sottojoidea nell’arco di 2 mesi accompagnata da crisi respiratorie anche a riposo. Effettuate radiografia del Rx torace TC collo si evidenzia una importante deviazione tracheale con stenosi del diametro di 7mm nel punto di maggiore compressione. L’estensione caudale del gozzo risulta all’arco aortico. Viene inizialmente sospettato un tumore anaplastico.L’approccio chirurgico mira alla esecuzione di biopsie tiroidee effettuate in teeam con gli otorinolaringoiatri che eseguono tracheostomia, al fine di bypassare il punto stenotico. Successivamente l’esame istologico mostrerà la presenza di un gozzo multinodulare.Pertanto viene posta l’indicazione alla tiroidectomia, eseguita da una equipe congiunta costituita dall’endocrinochirurgo e dall’otorinolaringoiatra,data la complessità dell’intervento per le aderenze conseguenti alla cannula tracheale e per la necessita di incisione più craniale rispetto alla norma. Al termine dell’intervento, persistendo la stenosi tracheale, è stato necessario il mantenimento della tracheostomia.Pertanto, si è reso necessario il posizionamento di una protesi a T di Montgomery da parte del chirurgo toracico.Dopo una settimana è stato possibile rimuovere la tracheotomia, mantenendo la protesi endotracheale, sostituita dopo 60 giorni con una protesi tubulare, che verrà rimossa tra circa 2-3 mesi. RISULTATI A tutt’oggi la paziente è tornata ad una vita regolare con normale funzione respiratoria senza l’ausilio di cannule esterne e soprattutto mantenendo una normale fonazione,senza alcun reliquato di natura chirurgica, che le ha permesso di tornare ad una qualità di vita perfettamente compatibile con la sua età CONCLUSIONI Da circa 10 anni l’approccio chirurgico standard alla patologia tiroidea è la rimozione dell’intera ghiandola anche per patologie benigne, al fine di evitare possibili reinterventi. Si preferisce tale atteggiamento radicale in quanto la terapia chirurgica conservativa effettuata nel passato ha reso necessari numerosi reinterventi su tiroidi già operate. Il nostro caso clinico è un esempio di complessità di patologia tiroidea recidiva che ha reso necessario un approccio multidisciplinare per la risoluzione delle complicanze derivanti da un reintervento.

This report describes the use of intraoperative parathyroid hormone (ioPTH) assay during parathyroidectomy for patients with secondary hyperparathyroidism on the waiting list for renal transplantation. The levels of ioPTH were determined among waiting list patients undergoing subtotal parathyroidectomy and tertiary hyperparathyroidism patients undergoing procedures. The levels of ioPTH were significantly reduced at 10 minutes by 59.7,3% among with secondary hyperparathyroidism and 68.9% among tertiary hyperparathyroidism. A 15 minutes it was 85% in secondary hyperparathyroidism and 89.7% in tertiary hyperparathyroidism. A decrement of 50% in basal values at 10 minutes and 85% decrement or more at 15 minutes was predictive for the success of abnormal parathyroid gland removal. The application of this technique during subtotal parathyroidectomy results was useful to predict a correct excision of abnormal parathyroid glands among patients with secondary hyperparathyroidism on the waiting list and for tertiary hyperparathyroidism patients.

Hemostatic disorders can often complicate transplantation procedures. Moreover, antihemmorhagic drugs may not efficiently control bleeding that occurs in such cases. We report on a patient who underwent kidney transplantation complicated by bone marrow aplasia and gastric bleeding who was succesfully treated with recombinant activated FVII (Novoseven). In May 2005, a 53-year-old man affected by chronic renal insufficiency underwent kidney transplantation. At the beginning of June, laboratory tests showed progressive reduction in the blood cell count with anemia, granulocytopenia, and thrombocytopenia related to the development of marrow insufficiency. We commenced transfusion therapy and administered hematologic growth factors. On June 3, 2005, the patient underwent surgical procedure to repair the abdominal wall. Two days thereafter, the postsurgical period was complicated by an episode of melena. The patient received additional treatment with packed red cells, platelets, and fresh-frozen plasma. The gastrointestinal bleeding continued until June 9, 2005, when therapy with recombinant activated FVII (Novoseven) was commenced at an initial dose of 90 μgr/kg. The first bolus did not significantly reduce the blood loss; it was therefore administered as a successive bolus at the same dosage that was able to stop bleeding. Endoscopic examination performed the day after showed the absence of the hemorrhagic lesion in the gastric mucosa. In the subsequent days, the need for transfusion was dramatically reduced with no episode of bleeding. At the same time, the laboratory and clinical findings of marrow insufficiency disappeared. Our case report showed that the use of a global antihemorrhagic factor, such as Novoseven, can successfully control gastrointestinal bleeding even in complicated patients despite failure of traditional antihemostatic therapy.