Maria Concetta Gioviale

Histological findings of the internal inguinal ring in patients having indirect inguinal hernia

BackgroundAiming 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.MethodsIn 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.ResultsIn 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.ConclusionThe 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.

Comparison between local and regional anesthesia in arteriovenous fistula creation.

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%, P<.05) in pulsatility index (PI) measured by Doppler ultrasound. In Group B, PI and venous dilation remained unaltered in the postoperative phase. No complications such as thrombosis or occlusion were encountered among patients who underwent BPB. Conclusions 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.

Electrophoresis of proteins and DNA on horizontal sodium dodecyl sulfate polyacrylamide gels

An inexpensive Plexiglas apparatus which allows a simple and rapid preparation of horizontal polyacrylamide gels of different dimensions for different purposes, is described. Preparation of such gels is as easy and rapid as agarose gel preparation, and polymerized polyacrylamide gels are used to fractionate proteins or small DNA fragments using a common horizontal electrophoretic tank. This apparatus was used to electrophoretically fractionate proteins or DNA for immuno-blot analyses, particularirly in the study of the allergenic response to Parietaria judaica pollen in senescence, for Southern-blot hybridizations and in the study of DNA polymorphisms.

Pancreatic islets from non-heart-beating donor pig: Two-layer preservation method in an in vitro porcine model

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.

Using Polymeric Scaffolds for Vascular Tissue Engineering

With the high occurrence of cardiovascular disease and increasing numbers of patients requiring vascular access, there is a significant need for small-diameter (<6 mm inner diameter) vascular graft that can provide long-term patency. Despite the technological improvements, restenosis and graft thrombosis continue to hamper the success of the implants. Vascular tissue engineering is a new field that has undergone enormous growth over the last decade and has proposed valid solutions for blood vessels repair. The goal of vascular tissue engineering is to produce neovessels and neoorgan tissue from autologous cells using a biodegradable polymer as a scaffold. The most important advantage of tissue-engineered implants is that these tissues can grow, remodel, rebuild, and respond to injury. This review describes the development of polymeric materials over the years and current tissue engineering strategies for the improvement of vascular conduits.

Towards an ideal source of mesenchymal stem cell isolation for possible therapeutic application in regenerative medicine

BACKGROUND The possibility of obtaining mesenchymal stem cells (MSCs) from fetal tissue such as amniotic fluid, chorionic villi and placenta is well-known and a comparison between MSCs originating in different sources such as fetal tissue and those from bone marrow in terms of yield and function is a topical issue. The mesenchymal stem cells isolated from bone marrow are well-characterized. Unfortunately the low quantitative yield during isolation is a major problem. For this reason, other tissue sources for MSCs are of paramount importance. CONCLUSION In this review, starting from a description of the molecular and cellular biology of MSCs, we describe alternative sources of isolation other than bone marrow. Finally, we describe the potential therapeutic application of these cells.

Abnormal espansion of segmented filamentous bacteria in the gut: a role in pathogenesis of chronic in fiammatory intestinal desease

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 Crohns 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.

Is secondary hyperparathyroidism-related myelofibrosis a negative prognostic factor for kidney transplant outcome?

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.

Isolation and Culture of β-Like Cells From Porcine Wirsung Duct

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 <3.2%, for glucagon. The positive ratio of immunoreactive staining was dependent on the glucose concentration; 17.8 mmol/L glucose effectively stimulated insulin- and glucagon-secreting cells. We concluded that porcine Wirsung duct cells were capable of proliferation with the potential to differentiate toward beta cells upon glucose induction in vitro.

Use of Intraperitoneal ePTFE Gore Dual-Mesh Plus in a Giant Incisional Hernia After Kidney Transplantation: A Case Report

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 correctly 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.