Proof-of-concept validation of the mechanism of action of Src tyrosine kinase inhibitors in dystrophic mdx mouse muscle: in vivo and in vitro studies.

Src tyrosine kinase (TK), a redox-sensitive protein overexpressed in dystrophin-deficient muscles, can contribute to damaging signaling by phosphorylation and degradation of ß-dystroglycan (ß-DG). We performed a proof-of-concept preclinical study to validate this hypothesis and the benefit-safety ratio of a pharmacological inhibition of Src-TK in Duchenne muscular dystrophy (DMD). Src-TK inhibitors PP2 and dasatinib were administered for 5 weeks to treadmill-exercised mdx mice. The outcome was evaluated in vivo and ex vivo on functional, histological and biochemical disease-related parameters. Considering the importance to maintain a proper myogenic program, the potential cytotoxic effects of both compounds, as well as their cytoprotection against oxidative stress-induced damage, was also assessed in C2C12 cells. In line with the hypothesis, both compounds restored the level of ß-DG and reduced its phosphorylated form without changing basal expression of genes of interest, corroborating a mechanism at post-translational level. The histological profile of gastrocnemius muscle was slightly improved as well as the level of plasma biomarkers. However, amelioration of in vivo and ex vivo functional parameters was modest, with PP2 being more effective than dasatinib. Both compounds reached appreciable levels in skeletal muscle and liver, supporting proper animal exposure. Dasatinib exerted a greater concentration-dependent cytotoxic effect on C2C12 cells than the more selective PP2, while being less protective against H2O2 cytotoxicity, even though at concentrations higher than those experienced during in vivo treatments. Our results support the interest of Src-TK as drug target in dystrophinopathies, although further studies are necessary to assess the therapeutic potential of inhibitors in DMD.

Evaluation of potential synergistic action of a combined treatment with alpha-methyl-prednisolone and taurine on the mdx mouse model of Duchenne muscular dystrophy.

AIMS: Glucocorticoids are the sole drugs clinically used in Duchenne muscular dystrophy, in spite of the relevant side effects. Combination of glucocorticoids with synergistic drugs may be one strategy to lower doses and control side effects, meanwhile providing wider control of the complex pathology. This study is a preclinical evaluation of the effect of a combined treatment of α-methyl-prednisolone (PDN) with taurine, a safe aminoacid with positive effects on some pathology-related events. METHODS: PDN (1 mg/kg/day i.p.) and taurine (1 g/kg/day orally) were administered either alone or in combination, for 4-8 weeks to male dystrophic mdx mice chronically exercised on a treadmill. Effects were assessed in vivo and ex vivo with a variety of methodological approaches. RESULTS: In vivo, each treatment significantly increased fore limb strength, a marked synergistic effect being observed with the combination PDN + taurine. Ex vivo, PDN + taurine completely restored the mechanical threshold, an electrophysiological index of calcium homeostasis, of extensor digitorum longus myofibres and the benefit was greater than for PDN alone. In parallel, the overactivity of voltage-independent cation channels in dystrophic myofibres was reduced. No effects were observed on plasma levels of creatine kinase, while lactate dehydrogenase was decreased by taurine and, to a minor extent, by PDN + taurine. A similar histology profile was observed in PDN and PDN + taurine-treated muscles. PDN + taurine significantly increased taurine level in fast-twitch muscle and brain, by high-pressure liquid chromatography analysis. CONCLUSIONS: The combination PDN + taurine has additive actions on in vivo and ex vivo functional end points, with less evident advantages on histopathology and biochemical markers of the disease.

[A 7-case anatomopathology revision in the presence of renal relapse after conservative therapy: implications on surgical technique].

Anatomopathology revision of the cases which underwent second surgery because of a renal neoplasm relapsing after conservative surgery, in order to find possible relations with the surgical technique. MATERIALS AND METHODS. At our institution, Nephron-sparing surgery (NSS) is currently indicated as elective technique for neoplasms smaller than 4cm in diameter. The technique involves the removal of the neoplasm with a margin of healthy parenchyma and with the perilesional fat. The patients are monitored with a first CT check after 4 months and then with ultrasound/CT checks every 6 months in the first 2 years and then once a year. In the present study we analyze the records of the cases in the period 1994-2005 undergoing a second operation for a renal tumor relapsing in the operated kidney after NSS. All specimens were reviewed by a single experienced uro-pathologist, who determined the size of the surgical margins and the relations between the seat of recidivism and the seat of the preceding enucleoresection. RESULTS. Seven cases with renal relapse were found out of 267 undergoing conservative surgery in the same period (incidence: 2.6%). The diagnosis had always been made lacking any other disease localizations at a complete re-staging; the average relapse latency was 19.4 months (8-46 months). In 5 cases the second tumor was found in the seat of the previous NSS: for these cases the minimum margin of the enucleoresection was lower than 3mm (median minimum margin: 1.6 mm). Differently, in the remaining 2 cases, both with a wider surgical margin (median minimum margin: 12.0 mm), the seat of the first and that of the second neoplasm were distant. In particular, in one case a multifocal relapse with a spread microvascular embolization was found, while in the other the two neoplasms showed a different histotype. DISCUSSION AND CONCLUSIONS. In the 5 cases with a little resection margin and relapsing tumor in the seat of the enucleoresection, the persistence of a peritumoral microscopic neoplastic disease can be assumed. In the other 2 cases showing a wider surgical margin the relapse can be attributed to the widespread microscopic multifocality in one case, and to the development of a second de novo neoplasm in the other case. The extension of the surgical margin seems then to have played a role in determining a relapse in the seat of enucleoresection.

[Nephron-sparing surgery versus radical nephrectomy in the treatment of renal cell carcinoma up to 7 cm].

To compare the oncological outcome of nephron-sparing surgery versus radical nephrectomy in renal cell carcinoma up to 7 cm by retrospectively reviewing our surgical experience. MATERIALS AND METHODS. Data collected from 1290 consecutive patients, who underwent surgery for renal carcinoma, have been stored since 1983 into a dedicated database. The cases with unilateral carcinoma up to 7 cm, pT1a/pT1b/pT3a N0/Nx M0, followed up for a minimum of 12 months if disease-free were reviewed. RESULTS. 732 patients were selected (mean follow-up: 72 months); 329 had a tumor less than 4 cm in diameter (182 cases of nephron-sparing surgery, 147 cases of nephrectomy), while for 403 of them the tumor was 4 cm or more (57 cases of nephron-sparing surgery, 346 cases of nephrectomy). The comparison between tumors less and equal to/more than 4 cm showed worse progression and disease-free survival rates for the latter, even though the type of surgery (nephron-sparing or radical) had no significant impact. Patients with extracapsular carcinoma ≥4 cm, treated with nephron-sparing surgery, had a particularly poor prognosis. CONCLUSIONS. The conservative management can be cautiously suggested for renal cancers up to 7cm, since the prognosis worsens proportionally with the diameter increase, with no statistical difference for both nephron-sparing and radical surgery. Nephron-sparing surgery proved to be the suitable treatment modality also for pT3a tumors measuring <4 cm, whereas when the tumor size increases, an adequate intraoperative evaluation of peritumoral tissues is essential to rule out fat infiltration. These results comply with the few similar studies available in literature, and suggest the possibility of designing a prospective study aiming at comparing conservative and radical surgery in the management of renal carcinoma up to 7 cm.

The collecting duct carcinoma of the kidney: a cytogenetical study.

OBJECTIVES: The Heidelberg classification of renal tumours identifies five histotypes of renal cancer, underlining for two of them (conventional and papillary renal cancers) a strict relation between the morphological aspect and the complement of alterations evidenced by the cytogenetic analysis of the neoplastic karyotype. Due to its low incidence, the collecting duct carcinoma (CDC) has not yet been characterized from a cytogenetic point of view. This study analyses the clinical, morphologic and cytogenetic features of the CDC observed and treated in our department. METHODS: From January 1995 to December 2002, among the 591 patients who underwent surgery for renal cancer, we observed 11 cases of CDC (prevalence 1.9%) treated either by radical (9 cases) or partial nephrectomy (2 cases). During radical nephrectomy a loco-regional lymphadenectomy was always performed. In the 9 cases observed after 1997, a complete cytogenetic analysis of the neoplastic karyotype was carried out. RESULTS: At pathological examination the disease was found to be confined to the renal capsule (TNM 1997 stage 1) in only 3 patients; venous neoplastic trombosis and nodal metastasis were present in 3 and 6 cases respectively; 2 patients showed distant metastases (lung, bone). Two of the patients affected with stage 1 tumours are still alive with no evidence of the disease at 48 and 88 months after surgery, while the third died following the systemic progression of a concomitant bladder carcinoma. One patient with stage 4 tumour (no. 11) is alive, but the follow up time is still limited (2 months). All the other 7 patients are dead after a mean survival time of 16.3 months (range 0-45). As for cytogenetic analysis, 2 CDCs didn't grow in culture and in one case no karyotype alterations were reported. In the remaining 6 cases hypodiploid stemlines and a homogeneous chromosome alteration pattern were observed, with multiple numerical and structural aberrations (mean 11.1, range 7-15) and the continuous involvement of chromosomes 1 and X or Y, both as traslocation and deletion/monosomy. Additional abnormalities of chromosomes 22 and 13 were found to be common but less frequent. CONCLUSIONS: The clinical behaviour of the CDC is aggressive and its prognosis is surely poor; surgical treatment seems to be curative only for organ-confined cancer, accounting for the minority of cases. This neoplasm is cytogenetically characterized by hypodiploid stemlines with common involvement of chromosome 1 and the autosomes.

Multitechnical pathological diagnosis in chromophobe renal cell carcinoma.

Two new cases of chromophobe renal cell carcinoma were diagnosed on the basis of their morphology and their karyotype complemented by flow cytometry. In one of these cases, however, all these investigations were not sufficient and additional histochemistry investigation had to be used to completely rule out other renal tumors such as oncocytoma, the prognosis of which is totally different.

[Conservative therapy of renal parenchymal neoplasia]. / Terapia conservativa della neoplasia renale parenchimale.

We review our overall experience in 1375 patients, who underwent surgery for renal cell carcinoma in the Departments of Urology of Brescia and Bergamo from 1983 to 1996. 185 (13.4%) patients had nephron-sparing surgery: imperative procedure was performed in 74 cases, while an elective surgery was done in 111 patients. Three years minimal follow up was considered in order to evaluate the outcome of surgical treatment in 48 patients who underwent imperative nephron-sparing surgery and in 73 with an elective procedure. Disease specific survival was 80.8% in the first group and it was 97% in the latter.

[Vascular and lymphatic anatomy of the penis]. / Cenni di anatomia emo-linfatica del pene.

Anatomic considerations of the penis and its lymphatic drainage are analyzed in this issue. The precise anatomic location of the inguinal and iliac lymph nodes and the fascial planes of the femoral canal is very important to the surgeon to reduce the high morbility of lymphadenectomy.

Strategies for Helicobacter pylori eradication in 1995: a review of international and Belgian experience.

After a decade of research, Helicobacter pylori eradication is still a problem because of the steady increase of bacterial resistance (imidazole, macrolides), pH-dependent efficiency of antibiotics, poor compliance of patients and frequent side effects of the therapies. After the failure of various monotherapies and the unefficiency of Amoxicillin-Imidazole combination for Imidazole-resistant strains, the two weeks ¿Oral Triple Therapy' with a 85% mean eradication rate, was abandoned because of a mean 35% side effects rate. The current goal is to obtain 90% eradication rate and the excellent results of german studies with a 2 weeks regimen combining a Proton Pump Inhibitor (PPI) with Amoxicillin have not been confirmed elsewhere in Europe. PPI plus Clarithromycin (two weeks) gave a mean 72% eradication rate on an ITT basis. The short, low-dose combination PPI-Clarithromycin-Imidazole for one week proposed by Bazzoli is very efficient in a population where Imidazole resistant strains are rare. The recent result of one week with (Omeprazole 20-Clarithromycin 250-Tinidazole 500) BID or (Omeprazole 20-Clarithromycin 500-Amoxi 1000) BID reached a 95% eradication rate but these very promising results are not confirmed in Belgium in an on-going study including 147 patients.

Surgery of lung metastases in renal cell carcinoma.

OBJECTIVE: To examine the impact on survival of pulmonary resection in the treatment of renal cell carcinoma that has metastasized to the lung. PATIENTS AND METHODS: From June 1986 to July 1991, 19 patients were submitted to synchronous or asynchronous lung metastasectomy. Histological examination confirmed the presence of lung metastases from renal cell carcinoma in 16 cases (six synchronous and 10 asynchronous with a mean disease-free interval of 23 months). In the other three cases, histological examination revealed tuberculomas, chondroid hamartoma and foci of anthracosis. RESULTS: To date, among the six patients with synchronous lung metastases, three have died, two are progressing and one has no evident disease after a mean survival of 24 months. Among the 10 patients who underwent surgical resection of metachronous lung metastases, one has died, three are progressing and six have no evident disease (mean survival time, 43 months). CONCLUSION: While the presence of synchronous lung metastases is an unfavourable prognostic factor even after surgical removal (five out of six patients died or are in progression shortly after metastasectomy), the results after surgery of asynchronous lung metastases are encouraging, although the real efficacy of this treatment is still to be confirmed.

[Ureteral malformations in the adult]. / Malformazioni ureterali nell'adulto.

Ureteral malformations that concern adults are very few. The aim of this review is to value the real clinical incidence and to illustrate etiology, diagnosis and therapeutical management advised in the literature.

[Controversies concerning the treatment of ureteral complications following kidney transplantation]. / Controversie nel trattamento della patologia dell'uretere trapiantato.

The ureteral complications after renal transplantation are urine leakage, stenosis and vesicoureteral reflux. The treatment is influenced by immunosuppression and difficult surgery (for bleeding and fibrosis). We report 8 cases with ureteral complication after renal transplantation. Stenosis were present in 5 cases: we performed ureterocystoneostomy by Politano-Leadbetter technique in 4 and pyelocystoanastomosis in 1. Vesicoureteral reflux were present in 3 cases: we preformed ureterocystoneostomy by Politano-Leadbetter technique in 2 and endoscopic infiltration with teflon of ureterovesical junction in 1. At present all patients have a normal renal function and absence of urinary tract infection.

Colonoscopic snare polypectomy: analysis of 1485 resections comparing two types of current.

The complications related to 1485 colonic snare polypectomies were analyzed according to the type of current which was used for resection. From January 1982 to August 1986 (period 1), blended current was used in 758 snare polypectomies, while continuous coagulation current was applied in 727 polypectomies from September 1986 to October 1989 (period 2). The incidence of complications observed in the entire series was low, consisting of 0.26% perforations and 0.9% major hemorrhages. There were no deaths in this series. Twelve of 14 hemorrhages occurred after resection of polyps larger than 1 cm. Although the incidence of complications was not statistically different in the two groups, there was a significant difference in the timing of hemorrhages. All of the major hemorrhages were immediate (eight) when the blended current was used, but delayed (six; from 2 to 8 days after polyp resection), when pure coagulation current was applied.

[Our experience in surgery of metastases of parenchymal renal neoplasms: mean follow-up time results]. / Nostra esperienza nella chirurgia delle metastasi da neoplasia renale parenchimale: risultati a medio termine.

The Authors report their experience about surgical treatment of metastases from renal cell carcinoma. From January 1983 to December 1989, twenty eight patients, 19 males and 9 females, with a median age of 58 years (range 42-79), have been submitted to contemporary or subsequent metastasectomy. Metastases were synchronous in 10 cases, while they appeared after a free disease mean-time of 28 months after nephrectomy in 18 patients. Recovery period has always been normal and all the patients were controlled, every six months, with routine blood and urine examinations, Chest X-ray, abdominal CAT and bone scan. Among the 10 patients with synchronous metastases 5 died, 3 are in progression and 2 are NED after a mean-time follow up of 36 months. Among 18 patients who underwent surgery for metachronous metastases, 2 died, 4 are in progression and 12 NED (mean follow up of 36 months). In conclusion, while the presence of synchronous metastases is an unfavourable prognostic factor even after their surgical removal (8 out of 10 patients died or are in progression shortly after metastasectomy), results after metachronous metastases surgery are encouraging, but the real efficiency of this treatment is still to be confirmed.

[Role of postoperative adjuvant radiotherapy in the treatment of class T2-3 NO MO adenocarcinoma of the kidney]. / Ruolo della radioterapia postoperatoria adiuvante nel trattamento dell'adenocarcinoma renale classe T2-3 N0 M0.

From 1968 to 1983, in the Department of Urology and in the Institute of Radiotherapy of the University of Brescia, the role of postoperative radiotherapy (PORT) in 95 patients with renal adenocarcinoma (T2-3 N0 M0) was investigated. From 1968 to 1978, 46 patients underwent radical nephrectomy and PORT; from 1978 to 1983, 49 patients were submitted to radical nephrectomy with regional lymphadenectomy (CH). Overall survival (PCS) at 5 years is 63% (PORT) vs 57% (CH) (p greater than 0.05). The probability of survival is better for left-sided neoplasms than for right-sided ones. In the CH group, the 5-year PCS is 40% vs 70%, respectively, for right vs left neoplasms (p less than 0.05); in the PORT group, PCS is 59% (right) vs 70% (left). For right-sided cancers, 5-year PCS is higher for PORT (59%) than for CH (40%) patients (p less than 0.05). In the PORT group acute bowel toxicity was 24% (grade 2, WHO). In 2 patients only (4.3%) PORT was stopped because of toxicity. PORT sequelae were investigated in: spinal cord, contralateral kidney, liver and bowel. Bowel sequelae (grade 2, Dische) were observed in 3 patients only (6.5%). In the T2-3 N0 M0 classes, radical nephrectomy with PORT may give the same results as aggressive surgery, with a low biological cost. Prognostic data might mean a different and more favorable loco-regional evolution for left-sided renal cancers.

[Bladder neck cysts: role of transrectal echography]. / Cisti del collo vescicale: ruolo dell'ecografia transrettale.

High frequency probes for transrectal sonography are well tolerated and permit us to study the cervico-urethral unit carefully. We are allowed to single out some rare causes of obstruction, otherwise difficult to diagnose. Bladder neck obstruction secondary to a cyst is one of these. Our case report is about a young patient with complete retention secondary to bladder cyst discovered by transrectal sonography.

[Surgical treatment of caval thrombosis caused by parenchymal renal neoplasms]. / Trattamento chirurgico della trombosi cavale da neoplasia renale parenchimale (N.R.P.).

The incidence of renal cell carcinoma with a vena caval tumour thrombus has been reported in the literature, form 4% to 19%. Vena caval involvement causes serious diagnostic and therapeutic problems. Surgical treatment is usually conditioned by the tumor thrombus cranial extension and the possible invasion of the vena caval wall. Using Diagnostic Imaging (ECHO, CAT, MRI) we are able to establish the real presence, dimension and extension of the tumor thrombus, but we can not evaluate precisely its nature or the infiltration of the vena caval wall. We report our own experience in 27 patients with renal cell carcinoma extending into the vena cava (22 cases with tumor thrombus extending under the diaphragm and 5 cases over the diaphragm) and describe our favourite approach for thrombus extending into the right atrium using extracorporeal circulation, profound hypothermia and cardiac arrest (3 cases). From our data, we believe that the vena cava involvement doesn't make the prognosis any worse, if it isn't associated with the infiltration of the vena caval wall and nodal disease.

Adjuvant medroxyprogesterone acetate to radical nephrectomy in renal cancer: 5-year results of a prospective randomized study.

From July 1, 1979 to June 30, 1983, 136 consecutive patients from 5 centers in Lombardy entered a prospective randomized study to compare 500 mg. adjuvant medroxyprogesterone acetate 3 times a week for 1 year to no treatment following radical nephrectomy for category M0 renal cancer. After a median followup of 5 years (range 42 to 90 months) 40 of 120 evaluable patients (33.3 per cent) experienced relapse after a median interval free of disease of 17 months (range 2 to 74 months). Relapses occurred in 19 of 58 evaluable patients in the adjuvant treatment group (32.7 per cent) and in 21 of the 62 evaluable controls (33.9 per cent). Sex steroid hormone receptors were studied in 102 of the 120 evaluable patients with the dextran-coated charcoal technique. No significant correlation could be found among receptors, relapses and treatment. On the other hand, 33 (56.9 per cent) of the 58 treated patients experienced 39 complications related to the long-term hormonal therapy. Three patients had to discontinue medroxyprogesterone acetate for severe toxicity after 2 to 3 months. Medroxyprogesterone acetate cannot be recommended as adjuvant therapy to radical nephrectomy in patients with renal cell carcinoma.

[Hormone receptors and endocrine therapy of renal carcinoma]. / Recettori ormonali ed endocrinoterapia nel carcinoma renale.

One hundred sixty-five consecutive patients with resectable renal cancer entered a cooperative study to evaluate hormone treatment and steroid receptors. Twenty-nine patients with concomitant distant metastases (category M1) received intramuscular medroxyprogesterone acetate (MPA) 500 mg/day for at least two months after the operation. No measurable remission was observed, but 8 of 24 evaluable patients (33%) had disease stabilization for a median duration of 6 months. One hundred thirty-six cases with category M0 cancer were randomly allocated to a control group or to a treatment group with MPA 500 mg/3 times a week for one year. After a median follow-up period of over 3 years, 30 of 121 evaluable patients (24.8%) had a relapse, usually in distant sites. Relapses and survival were independent from postoperative treatment and sex. Only the extent of the disease and the presence of steroid receptors in the tumor were related with prognosis, but no relation could be found between receptors and response to hormone treatment. The presence of low concentrations of hormone receptors in a proportion of renal cancers remains unclear. However, MPA is confirmed to be only marginally active in metastatic renal cancer and the drug cannot be recommended as adjuvant to radically resected patients because of significant toxicity and lack of therapeutic activity.