Biological effects of low frequency high intensity ultrasound application on ex vivo human adipose tissue.

In the present work the effects of a new low frequency, high intensity ultrasound technology on human adipose tissue ex vivo were studied. In particular, we investigated the effects of both external and surgical ultrasound-irradiation (10 min) by evaluating, other than sample weight loss and fat release, also histological architecture alteration as well apoptosis induction. The influence of saline buffer tissue-infiltration on the effects of ultrasound irradiation was also examined. The results suggest that, in our experimental conditions, both transcutaneous and surgical ultrasound exposure caused a significant weight loss and fat release. This effect was more relevant when the ultrasound intensity was set at 100 % (~2.5 W/cm², for external device; ~19-21 W/cm2, for surgical device) compared to 70 % (~1.8 W/cm² for external device; ~13-14 W/cm2 for surgical device). Of note, the effectiveness of ultrasound was much higher when the tissue samples were previously infiltrated with saline buffer, in accordance with the knowledge that ultrasonic waves in aqueous solution better propagate with a consequently more efficient cavitation process. Moreover, the overall effects of ultrasound irradiation did not appear immediately after treatment but persisted over time, being significantly more relevant at 18 h from the end of ultrasound irradiation. Evaluation of histological characteristics of ultrasound-irradiated samples showed a clear alteration of adipose tissue architecture as well a prominent destruction of collagen fibers which were dependent on ultrasound intensity and most relevant in saline buffer-infiltrated samples. The structural changes of collagen bundles present between the lobules of fat cells were confirmed through scanning electron microscopy (SEM) which clearly demonstrated how ultrasound exposure induced a drastic reduction in the compactness of the adipose connective tissue and an irregular arrangement of the fibers with a consequent alteration in the spatial architecture. The analysis of the composition of lipids in the fat released from adipose tissue after ultrasound treatment with surgical device showed, in agreement with the level of adipocyte damage, a significant increase mainly of triglycerides and cholesterol. Finally, ultrasound exposure had been shown to induce apoptosis as shown by the appearance DNA fragmentation. Accordingly, ultrasound treatment led to down-modulation of procaspase-9 expression and an increased level of caspase-3 active form.

Effects of phosphatidylcholine and sodium deoxycholate on human primary adipocytes and fresh human adipose tissue.

Recent studies introduced the novel concept of chemical lipolysis where phosphatidylcholine (PC), an active component of commercial preparations, plays a pivotal role. Other studies suggested that sodium deoxycholate (DOC), an excipient contained in medical preparations, could be the real active component performing an adipocytolytic action. We investigated the effects of PC and DOC on human primary adipocyte cultures and on human fresh adipose tissue. Human adipocytes isolated by Rodbell's method, were cultured onto type I collagen-coated glass coverslips, placed into 24-well tissue culture plates. Cells were incubated with or without DOC (5-7-9%), PC (5%) or DOC/PC mixture and observed under phase contrast microscope. After incubation, cells were stained with Oil Red-O and with acridine orange/ethidium bromide to observe necrotic cells with phase contrast microscope and fluorescent microscope, respectively. Histological specimens from adipose tissue biopsies were observed with phase contrast microscopy and with scanning electron microscopy. To investigate the lipid pattern variability in the different experimental conditions, culture medium obtained from the different treatments was subjected to lipid extraction and subsequently to thin layer chromatography (TLC). Microscopic observation of adipocytes showed that DOC treatment led to a detrimental morphological effect in a dose-dependent manner. PC treatment did not significantly affect adipocyte viability. On the contrary, results from experiments aimed to analyze the effects of PC/DOC combined treatment suggested a PC protective role against the DOC harmful effects on adipocytes. Results indicated that clinical effects, observed in local treatment with pharmaceutical preparation, could be due only to DOC, a detergent inducing nonspecific lysis of cell membranes following adipocyte necrosis. On the other hand, PC could likely be incorporated in the lipid bilayer, thus strongly reducing the disruptive DOC effects.

[Complications of elective hepatic resections. Analysis of risk factors]. / Le complicanze della chirurgia resettiva epatica d'elezione. Analisi dei fattori di rischio.

In the last decades the hepatic surgery has been more and more employed thanks to improvement of the surgical technique and of the post-operative assistance which have brought the peroperative mortality of principal specialistic centres to less of 5%. The main post-operative complications which trouble the hepatic surgery, forming in same cases the cause of the death, are: hepatic insufficiency, haemorrhage, subphrenic abscess and the appearance of biliary fistulas. These complications are often connected and linked to the devitalization of a part of the residual parenchyma. We have made a retrospective study on a series of 214 hepatic resections, executed in election, to estimate the main pre and intra-operative risk factors. The operative mortality has been zero whereas the post-operative one is occurred in 4.2% of the cases with a morbidity of 27.5%. In our experience the meaning full factors to prefigure an operative risk are resulted: the associated pathologies like diabetes, cardiopathies, ipertension and bronchopathies; the length of the operation; the entity of the peroperative haematic loss and of the consequent transfusional therapy and eventually quality the residual parenchyma.

Surgical treatment of hepatic metastatic colorectal cancer.

73 patients (men age 54 years, range 17-78), underwent an hepatic resection for metastatic, colorectal cancer. Operative mortality was 1.36% Overall 5 years survival rate (Kaplan-Meier) was 27%; 5 years disease free interval was equally 27%. This could demonstrate that 5 years survivors could be also considered free from the risk of metastatic recurrence. This is confirmed by our survivors over 5 years (70-79-94 months) that are still disease free.

[Proctocolectomy and mechanical ileo-anal anastomosis without mucosectomy]. / Proctocolectomia e anastomosi meccanica ileo-anale senza mucosectomia.

The present study compared the outcome in a small series of patients (7 cases) who underwent total proctocolectomy without mucosal proctectomy and stapled ileal pouch-anal anastomosis, constructed at the apex of the anal transitional zone, with our previous experience (17 cases) in which the ileal pouch was anastomosed at the dentate line after mucosectomy. Though not statistically significant, our limited experience showed excellent clinical results with better continence and discriminating ability between gas and faeces in the former group. The resting anal pressure profile showed no chances in the postoperative period. The operation time was significantly reduced compared with our previous approach which was a time-consuming procedure. Furthermore, a reduced risk of complications (pelvic sepsis, haemorrhage) was observed.