Antonio Di Cataldo

BENEFICIAL EFFECTS OF RUTIN AND L-ARGININE COADMINISTRATION IN A RAT MODEL OF LIVER ISCHEMIA/REPERFUSION INJURY

Reperfusion following liver ischemia results in oxidative stress leading to liver injury. The aim of this study was to investigate the combined effects of two antioxidant agents, rutin and L-arginine, in rat liver ischemia-reperfusion (I/R). Male Wistar rats were divided into five groups: 1) sham operated, 2) I/R, 3) I/R+rutin, 4) I/R+L-arginine, and 5) I/R+rutin+L-arginine. Plasmatic and hepatic levels of alanine transaminase (ALT), aspartate transaminase (AST), lipid peroxides (LOOH), and thiol groups (RSH) were examined, as well as DNA fragmentation and liver histopathology. Furthermore, to elucidate the pathophysiological processes involved in the antioxidant mechanism(s) of rutin and L-arginine, we assessed the expression of inducible (iNOS) and endothelial nitric oxide synthase (eNOS) isoforms and heme oxygenase-1 (HO-1), both playing key roles in the biochemical cascade of liver injury. Significant increase in plasmatic ALT and AST activities were observed in untreated I/R rats compared with sham-operated animals, whereas treatment with rutin or L-arginine in I/R rats reduced hepatic damage. Interestingly, combined therapy with rutin and L-arginine resulted in a further reduction of plasmatic ALT and AST activities compared with rutin or L-arginine alone. These results were further confirmed by the analysis of DNA fragmentation, LOOH, RSH groups, and liver histopathology, which showed the highest protective effects following the coadministration of rutin and L-arginine. Finally, the combined therapy protocol resulted in a significant induction of liver HO-1 and a concomitant reduction of iNOS expression that may both be responsible for the beneficial effects of the proposed pharmacological protocol.

Cytotoxic and immune-sensitizing properties of nitric oxide-modified saquinavir in iNOS-positive human melanoma cells†‡

We have recently shown that covalent attachment of the NO moiety to the HIV protease inhibitor Saquinavir (Saq) produced a qualitatively new chemical entity, named Saquinavir‐NO (Saq‐NO), with enhanced anticancer properties and reduced toxicity. In this study we evaluated the impact of Saq‐NO on the growth of A375 human melanoma cells, as a prototype of NO‐dependent cancer model. The novel compound strongly affected the in vitro and in vivo progression of A375 melanoma cell growth. The mechanism of antimelanoma action comprised dual drug activity—induction of apoptotic cell death and acquisition of melanoma cell responsiveness to TRAIL. Saq‐NO‐triggered apoptosis was dependent on transient AKT up‐regulation and reduced pERK and iNOS expression that were observed within the first 12 h of exposure to the drug. Thereafter, however, Saq‐NO up‐regulated both iNOS transcription and NO endogenous synthesis and sensitized A375 cells to TRAIL. Furthermore, reduced YY1 expression was observed after 24 h of Saq‐NO exposure, which correlated with increased expression of DR5. The biological relevance of this complex and powerful action of Saq‐NO was consistent with the marked drug‐induced inhibition of the growth of A375 xenotransplants in nude mice. J. Cell. Physiol. 226: 1803–1812, 2011.

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

Monitoring carcinoembryonic antigen in colorectal cancer: Is it still useful?

The results of a study conducted to determine the usefulness of carcinoembryonic antigen (CEA) monitoring in the follow-up of patients with resected colorectal cancer are reported herein. The subjects of this study were 125 patients in whom CEA had been determined preoperatively and 239 patients in whom CEA had been monitored postoperatively. The results revealed increased preoperative CEA in only 24% of the subjects, and that this increment was correlated with subsequent more advanced tumor stage and a higher recurrence rate (P<0.01). The postoperative CEA level exceeded the threshold in 71% of the patients affected by recurrence, 94.4% of whom developed liver metastases and 50%, nonhepatic recurrence. This marker showed elevated sensitivity for liver metastases (99%), whereas the sensitivity was lower for nonhepatic recurrence of the disease (94%). Thus, we concluded that CEA monitoring can be useful for preoperative colorectal tumor grading, even if its validity in the early diagnosis of recurrence is problematic, especially in terms of radical repeated surgery and survival.

Experimental models in microsurgery

The development of experimental microsurgery can be considered the natural evolution of a diffuse need to increase precision in many fields of surgery. Microsurgery accelerated the possibility of deepening many unclear aspects of pathophysiology, using miniaturized and reproducible experimental models. We report briefly on the fundamental principles of microsurgery and the most frequently performed and useful models of experimental microsurgery, especially to employ as training models for surgeons, but also as bases for developing new and always‐welcome models.

Usefulness of microsurgery in the training of the general surgeon

It is a common opinion that general surgery is the first step for whoever approaches a surgical discipline, and that whoever practices training in general surgery should learn the rudiments of each surgical branch. The role of microsurgery in the training of the general surgeon has not been well‐established. Clinical applications of microsurgery in general surgery are few and are rarely required, and have been connected strictly to restricted indications. However, we think that microsurgery could be very useful to the general surgeon because it allows the execution of experimental research on rats, the only possibility permitted by law. In these studies the microsurgeon can perform many times and in a short time the same surgical operation, thus improving his skill, and easily getting familiarity with surgical instruments and sutures.

Three microsurgical courses in Catania

The utilization of microsurgical techniques in the surgical disciplines is now accepted worldwide, and many surgeons should receive the opportunity of learning these techniques. To meet this requirement, microsurgical courses have been organized and comprise both theoretical aspects and, especially, practical demonstrations. A one‐week course usually allows attending surgeons to get familiar with magnification and to perform microvascular anastomoses, vasovasostomy, tubal reconstruction, sciatic nerve suture, and end‐to‐side portacaval shunt, a useful exercise before dealing with rodent organ transplantation. It is important to underline that the skill supplied by these microsurgical courses is not enough to start clinical applications of microsurgery; to this end, training must continue long after the course has been finished.

Colorectal Follow-Up Planning Modified on the Basis of Our Personal Experience

Background: The authors, evaluating the disappointing follow-up results in patients suffering from colorectal carcinoma who had undergone surgery for cure, tried a more rational follow-up. Methods: In a retrospective review about 232 patients who adhered to the follow-up protocol, we evaluated the accuracy rates of CEA, liver ultrasonography and abdominal CT. In the same group of patients, we evaluated the type of correlation between the neoplastic recurrence rate and Astler-Coller’s classification. Results: (1) In detecting hepatic metastases CEA levels furnished sensitivity and negative predictive value more than liver ultrasonography (83.3 vs. 77.8% and 98.4 vs. 98%, respectively); (2) in our series, we obtained a lower recurrence rate in classes A+B1 (7.5%) and B2 (20.8%) and higher in C1+C2 (44.4%) and D (66.7%) (p < 0.01). Conclusion: According to these data we decided to eliminate postoperative liver ultrasonography and customize follow-up protocol on tumor staging and timing of cancer relapse. The authors believe that these changes will not modify the results, but cause less psychophysical stress for the patients and reduce costs by 50%.

The Urachal Pathology with Umbilical Manifestation: Overview of Laparoscopic Technique

BACKGROUND The persistent patency of the urachus after birth is a rare anomaly, especially because most of the cases are asymptomatic. The guiding symptom for urachal-umbilical sinus and urachal cyst is the presence of umbilical discharge. Even if today we tend to laparoscopic treatment, in scientific literature there is still no evidence, because the reports are rare and often they are clinical cases. METHODS Thirteen patients with a symptomatic urachal pathology were evaluated; 12 of these were affected by umbilical discharge and 1 by periumbilical discomfort. Five of 13 were prospectively treated by laparoscopy and the remaining 8 patients, who had been previously treated with conventional surgery, formed the control group. The authors report the laparoscopic technique used, which allowed the complete excision of the urachus. RESULTS The operation time was lower for the patients treated by conventional surgery (71.9 minutes versus 101 minutes; P=.002), whereas the control pain (P=.05) and, above all, the excised urachus length (11.6 versus 8.7 cm; P=.03) were in favor of the patients treated by laparoscopic surgery. We registered only one recurrence in a patient treated by conventional surgery. CONCLUSION In the rare international scientific literature, only one study report comparative data, as our study. The results that we obtained seem to be in favor of the laparoscopic procedure, although prospective, randomized trials are needed to get stronger evidence.

Pancreas Transplantation in the Rat Using Lee's Technique: A Reliable Model in Experimental Microsurgery

Numerous unresolved problems, both technical and immunological, in pancreas transplantation stimulate experimental studies. Dogs have been routinely used in experimental studies but today rats are more commonly used. However, pancreas transplantation in the rat presents complex technical problems and requires a good knowledge of microsurgical techniques. In 1983 Squifflet undertook an experimental study aimed at evaluating the technical aspects of pancreas transplantation in the rat and calculating the success rates using different methods. The comparison of four methods revealed to our surprise that 100% of the rats operated on using Lees technique had complications, with a 0% survival rate. In our study we report our experience using Lees technique which we had the opportunity of mastering directly under the supervision of Professor Lee. We performed 100 pancreas transplantations using Lees technique and divided our study in two phases. In the first phase we performed 70 pancreas transplantations and overall survival, after 1 week, was 42 rats (60%). In the second phase on 30 rats diabetes was induced by administering 70 mg/kg of streptozotocin. These 30 diabetic rats underwent pancreas transplantation and overall survival, after 1 week, was 25 (83.3%). We believe that our successful survival rates could probably be explained by the close collaboration between Lee and our department. Moreover, we noted the importance of constant training in obtaining better results, and in our opinion Lees technique of pancreas transplantation is a reliable experimental model which can be used to resolve problems linked to pancreas transplantation.