Glial cell line-derived neurotrophic factor promotes invasive behaviour in testicular seminoma cells.

The glial cell line-derived neurotrophic factor (GDNF) has multiple functions that promote cell survival, proliferation and migration in different cell types. The experimental over-expression of GDNF in mouse testis leads to infertility and promotes seminomatous germ cell tumours in older animals, which suggests that deregulation of the GDNF pathway may be implicated in germ cell carcinogenesis. GDNF activates downstream pathways upon binding to its specific co-receptor GDNF family receptor-a 1 (GFRA1). This complex then interacts with Ret and other co-receptors to activate several intracellular signalling cascades. To explore the involvement of the GDNF pathway in the onset and progression of testicular germ cell tumours, we analysed GFRA1 and Ret expression patterns in seminoma samples. We demonstrated, via immunohistochemistry, that GFRA1, but not Ret, is over-expressed in in situ carcinoma (CIS) and in intratubular and invasive seminoma cells compared with normal human germ cells. Functional analysis of the GDNF biological activity was performed on TCam-2 seminoma cell line. Reverse transcription-PCR (RT-PCR) and immunohistochemical analyses demonstrate that TCam-2 cells express both GFRA1 and Ret mRNA, but only GFRA1 was detected at the protein level. In TCam-2 cells, although GDNF is not mitogenic, it is able to induce migration, as demonstrated by a Boyden chamber assay, possibly through the Src and MEK pathways. Moreover, GDNF promotes invasive behaviour, an effect dependent on pericellular protease activity, possibly through the activity of matrix metalloproteinases. GFRA1 over-expression in CIS and seminoma cells, along with the functional analyses in TCam-2 cells, suggests an involvement of the GDNF pathway in the progression of testicular germ cell cancer.

Completely laparoscopic versus open total gastrectomy in stage I-III/C gastric cancer: safety, efficacy and five-year oncologic outcome.

AIM: Laparoscopic gastrectomy is becoming a minimally invasive procedure widely accepted by laparoscopic surgeons; yet, many doubts remain about its oncologic efficacy in treating malignant neoplasia. Aim of this study was to analyze our experience comparing completely laparoscopic total gastrectomy to its laparotomic counterpart, about safety, efficacy and five-year oncologic outcome. METHODS: From January 2003 to October 2009, 25 patients with stage I-III/C gastric cancer (TNM Seventh Edition, 2010) were operated on and retrospectively compared to an homogeneous group of patients, stratified for age, stage of disease and comorbidities. Length of surgery, estimated blood loss, postoperative ileus, resumption of oral intake, morbidity, 30 days mortality, number of lymph nodes harvested, five years overall and disease free survival were analyzed, comparing the two groups. RESULTS: There was no conversion. Thirty days mortality was zero for both groups, while morbidity was 16% in the lap group, 32% in the open group (P<0.05). Length of operation was 211±23 min for the lap group, and 185±19 min for the open group (P>0.05); the estimated blood loss was 250±150 mL for the lap group, 495±190 mL for the open group (P<0.05). Number of lymph nodes harvested was 35±18 for the lap group, 40±16 for the open group (P>0.05). No port site metastatic implantation occurred in any patient treated laparoscopically; five years overall and disease free survival were 55.7% and 54.2% for the lap group, 52.9% and 52.1% for the open group, respectively, with no statistical difference (P>0.05). Completely laparoscopic total gastrectomy represents a new challenge for the laparoscopic surgeon. In spite of clear advantage for patients, some debate remains about its oncologic efficacy in the middle and long period, even if many authors report comparable results to open total gastrectomy. In our experience, it is a safe and valid alternative to its open counterpart, with no statistically different number of lymph nodes harvested, five years overall and disease free survival in respect to the open gastrectomy. Yet, it remains a complex procedure requiring high laparoscopic skill. CONCLUSION: In our opinion, completely laparoscopic total gastrectomy is a safe and effective procedure, with long term oncologic results not statistically different from the open procedure; yet, it requires high laparoscopic experience, especially to carry out an extended lymphadenectomy and to fashion the anastomosis. More randomized prospective trials are needed to state this procedure as a new gold-standard in treating stage I-III/C non metastatic gastric cancer.

Laparoscopic versus open total mesorectal excision for stage I-III mid and low rectal cancer: a retrospective 5 years analysis.

INTRODUCTION: Total mesorectal excision (TME) is the cornerstone of a correct surgical therapy for extraperitoneal rectal cancer. Aim of the study is to evaluate our 5 years experience confronting retrospectively laparoscopic (lap) TME in respect to its laparotomic (open) counterpart. PATIENTS AND METHODS: 30 patients were treated laparoscopically for stage I-III extraperitoneal rectal cancer and retrospectively compared to a homogeneous group, stratified for sex, age, comorbidities and stage of disease. RESULTS: 30 days mortality was zero for both groups, while morbidity was 20% for the lap group and 36.6% for the open group. Mean lymph nodes harvested was 24 ± 12 for the lap group, 26 ± 14 for the open group (p > 0.05). Five years overall and disease free survival was respectively 82.2% and 81.4% in the lap group, 79.9% and 79.6% in the open group, without statistical significance (p>0.05). Discussion. Minimally invasive TME resulted a safe, effective and oncologically adequate procedure when retrospectively compared to its laparotomic counterpart, with 5 years overall survival and disease free survival reaching no statistical significance compared to the open approach, but with all the advantages of the laparoscopy such as less pain and blood loss, faster recovery, less morbidity and better cosmetics. CONCLUSIONS: Our study has retrospectively demonstrated that laparoscopic TME is feasible and oncologically effective, even if it remains a complex minimally invasive procedure, requiring adequate skill. More prospective, randomized studies are necessary to define such a procedure as the new gold standard in treatment of stage I-III extraperitoneal rectal cancer.

Laparoscopic versus open appendectomy for the treatment of complicated appendicitis.

AIM: Laparoscopic Appendectomy (LA) is widely performed for the treatment of acute appendicitis. However the use of laparoscopic approach for complicated appendicitis is controversial, in particular because it has been reported an increased risk of postoperative IntraAbdominal Abscess (IAA). The aim of this study was to compare the outcomes of LA versus Open Appendectomy (OA) in the treatment of complicated appendicitis, especially with regard to the incidence of postoperative IAA. PATIENTS AND METHODS: A retrospective study of all patients treated at our institution for complicated appendicitis, from May 2004 to June 2009, was performed. Data collection included demographic characteristics, postoperative complications, conversion rate, and length of hospital stay. RESULTS: Thirty-eight patients with complicated appendicitis were analysed. Among these, 18 (47,3%) had LA and 20 (52,7%) had OA. There were no statistical differences in characteristics between the two groups. The incidence of postoperative IAA was higher (16,6%), although not statistically significant, in the LA compared with OA group (5%). On the other hand the rate of wound infection was lower (5%) in the LA versus OA (20%). CONCLUSION: Our study indicated that LA should be utilised with caution in case of perforated appendicitis, because it is associated with an increased risk of postoperative IAA compared with OA.

Laparoscopic versus open radical nephrectomy in T1-T2 renal carcinoma: personal 5-year experience about the oncologic outcome.

AIM: Since 1990 when it was firstly performed, radical laparoscopic nephrectomy has gained wide popularity because of its less morbidity and adequate oncologic outcome. The aim of this study was to report our experience about oncologic 5-year outcome of laparoscopic radical nephrectomy. METHODS: Fifteen patients were treated laparoscopically and retrospectively compared to a group of patients treated laparotomically, omogeneous for age, stage of disease and comorbidities. RESULTS: There was no conversion in the laparoscopic group and duration of both procedure showed no statistical difference. Laparoscopic procedures showed less intraoperative blood loss, less postoperative ileus, shorter hospitalization and less morbidity, all with statistical significance. Overall 5 years survival showed no statistical significant difference in the two groups (88.9% laparoscopic group vs. 86.2% laparotomic group). CONCLUSION: Laparoscopic radical nephrectomy has clear advantages compared to the traditional surgery, especially about less morbidity, less blood loss, shorter hospitalization, with an oncologic outcome absolutely comparable to the laparotomic procedure. Laparoscopic radical nephrectomy is a safe and oncologically adequate surgical procedure with clear advantages compared to the its open counterpart, so it must be considered as a valid alternative to laparotomic surgery in case of non metastatic T1-T2 kidney cancer.

[Totally laparoscopic total gastrectomy with esophago-jejunal termino-lateral anastomosis by Or-Vil device for carcinoma. Our experience in ten consecutive patients]. / Gastrectomia totale totalmente laparoscopica con anastomosi esofago-digiunale termino-laterale (sistema Or-Vil) nel trattamento del cancro gastrico: nostra esperienza su 10 casi consecutivi.

INTRODUCTION: Laparoscopic gastrectomy represents an alternative procedure for treatment of gastric cancer. Yet, some debate remains about its efficacy, basically from an oncologic point of view. Aim of this study is to analyze our experience with totally laparoscopic total gastrectomy with termino-lateral esophago-jejunal anastomosis by Or-Vil device. PATIENTS AND METHODS: From February 2007 to February 2008, 10 patients underwent the procedure with Or-Vil device for the esophago-jejunal termino-lateral anastomosis. ASA score, UICC-AJCC stage, number of lymph nodes harvested, mortality and morbidity were analyzed. RESULTS: All procedures were concluded laparoscopically. No mortality was observed, morbidity was 20%. Median of lymph nodes harvested was 30+/-14. No port implantations were observed. Discussion. Laparoscopic gastrectomy, both partial and total, is a new challenge, with clear advantages for the patients, but it still must demonstrate its efficacy, especially from the oncologic point of view. In our experience, we can state that totally laparoscopic total gastrectomy is safe, effective and oncologically correct; yet, it is technically demanding and more studies are required to confirm its oncologic efficacy when compared with laparotomic gastrectomy. CONCLUSIONS: Totally laparoscopic total gastrectomy with esophago-jejunal termino-lateral anastomosis by Or-Vil device represents, in our experience, a valid alternative to open procedure; yet, more prospective randomized trials are needed to define this procedure as a new standard for gastric cancer treatment.

A laser emitting contact lens for eye tracking.

In this paper, we present the first realisation and experimentation of a new eye tracking system using an infrared (iR) laser pointer embedded into a wireless smart contact lens. We denote this contact lens prototype as the cyclops lens, in reference to the famous hero of the X-Men comics. The full eye tracker device combines the smart contact lens and its eyewear, which provides a primary source of energy and the beam detection system. We detail the assembling and encapsulation process of the main functionalities into the contact lens and present how a gaze tracking system is achieved, compared to existing conventional eye-tracking ones. Finally, we discuss future technical improvements.

Metabotropic glutamate receptors regulate differentiation of embryonic stem cells into GABAergic neurons.

Mouse embryonic stem (ES) cells were stimulated to differentiate either as adherent monolayer cultures in DMEM/F12 supplemented with N2/B27, or as floating embryoid bodies (EBs) exposed to 1 microM retinoic acid (RA) for 4 days, starting from 4 DIV, and subsequently re-plated in DMEM/F12 medium. Adherent monolayer cultures of ES cells expressed mGlu5 receptors throughout the entire differentiation period. Selective pharmacological blockade of mGlu5 receptors with methyl-6-(phenylethynyl)-pyridine (MPEP) (1 microM, added once a day) accelerated the appearance of the neuronal marker, beta-tubulin. In addition, treatment with MPEP increased the number of cells expressing glutamate decarboxylase-65/67 (GAD(65/67)), a marker of GABAergic neurons. In floating EBs, mGlu5 receptors are progressively replaced by mGlu4 receptors. The orthosteric mGlu4/6/7/8 receptor agonist, L-2-amino-4-phosphonobutanoate (L-AP4), or the selective mGlu4 receptor enhancer, PHCCC,--both combined with RA at concentrations of 30 microM--increased the expression of both beta-tubulin and GAD(65/67), inducing the appearance of fully differentiated neurons that released GABA in response to membrane depolarization. We conclude that mGlu receptor subtypes regulate neuronal differentiation of ES cells in a context-dependent manner, and that subtype-selective ligands of these receptors might be used for the optimization of in vitro protocols aimed at producing GABAergic neurons from ES cells.

Geoarchaeological Evidence of Middle-Age Tsunamis at Stromboli and Consequences for the Tsunami Hazard in the Southern Tyrrhenian Sea.

Large-scale landslides at volcanic islands are one of the most dangerous geological phenomena, able to generate tsunamis whose effects can propagate far from the source. However, related deposits are scarcely preserved on-land in the geologic records, and are often difficult to be interpreted. Here we show the discovery of three unprecedented well-preserved tsunami deposits related to repeated flank collapses of the volcanic island of Stromboli (Southern Italy) occurred during the Late Middle Ages. Based on carbon datings, on stratigraphic, volcanological and archaeological evidence, we link the oldest, highest-magnitude investigated tsunami to the following rapid abandonment of the island which was inhabited at that time, contrary than previously thought. The destructive power of this event is also possibly related to a huge marine storm that devastated the ports of Naples in 1343 (200 km north of Stromboli) described by the famous writer Petrarch. The portrayed devastation can be potentially attributed to the arrival of multiple tsunami waves generated by a major landslide in Stromboli island, confirming the hypothetical hazard of these phenomena at a regional scale.

Recurrent episodes of syncope in a patient with cerebral arteriovenous malformation.

As there are a number of possible causes of syncope, differentiation between cardiovascular disease, neurogenic disease and other disorders is mandatory. Cerebral arteriovenous malformations (AVMs) are tangled anastomoses of blood vessels of varying calibre in which arteriovenous shunting occurs in a central nidus, which is the area towards which multiple feeding arteries converge and from which enlarged veins drain. We describe a clinical case of syncope caused by a large AVM discovered in a 66-year-old woman. The symptoms were probably related to an epileptogenic mechanism since the syncope disappeared following the administration of antiepileptic therapy. The anatomical, pathological and clinical aspects of AVMs are discussed.

[High-dose CEF (cyclophosphamide, epirubicin, fluorouracil) as primary chemotherapy in locally advanced breast cancer: long-term results]. / CEF (ciclofosfamide, epirubicina, fluorouracile) ad alte dosi come chemioterapia primaria nel trattamento del carcinoma mammario localmente avanzato: risultati a lungo termine.

PURPOSE: To determine wether primary CEF is effective in locally advanced breast cancer, as measured by response, local recurrences, disease free survival (DFS) and overall survival (OS). MATERIAL AND METHODS: From 1990 to 1998, 62 patients with stage III disease were enrolled into a prospective study at Regina Elena Institute for Cancer Research, Rome. Inflammatory breast cancer (IBC) was included. Patients received three 21 days cycles of chemotherapy that consisted in epirubicin 50 mg/m2, cyclophosphamide 400 mg/m2, and fluorouracil 500 mg/m2 i.v. on days 1 and 8. G-CSF (300 microg) was given subcutaneously every other day from day 5 to day 17. After primary chemotherapy, whenever possible, mastectomy or conservative surgery was performed. Subsequently responding patients received the same regimen, while non responders were given a non cross resistant chemotherapy. In case of conservative surgery or initial T4 tumor radiation therapy was performed at the end of adjuvant chemotherapy. ER positive patients received tamoxifen 20 mg/d for five years. RESULTS: Seven IIIA patients had a median OS of 43 months (C.I. 95%, 31-55) and DFS of 42 months (C.I. 95%, 16-68), while 15 IBC patients had a median OS of 52 months (C.I. 95%, 52-79) and DFS of 27 months (C.I. 95%, 14-39). Forty IIIB non inflammatory breast cancer patients had a median DFS of 87 months (C.I. 95%, 1-175); median OS was not reached. Ten-year OS was 28.6% for stage IIIA, 50.6% for stage IIIB and 36% for IBC. CONCLUSION: Primary CEF appear to be an effective treatment. In our study we obtained a good local control and interesting long term data of disease free and overall survival.

Cyto-histological and morpho-physiological responses of common duckweed (Lemna minor L.) to chromium.

Along with cadmium, lead, mercury and other heavy metals, chromium is an important environmental pollutant, mainly concentrated in areas of intense anthropogenic pressure. The effect of potassium dichromate on Lemna minor populations was tested using the growth inhibition test. Cyto-histological and physiological analyses were also conducted to aid in understanding the strategies used by plants during exposure to chromium. Treatment with potassium dichromate caused a reduction in growth rate and frond size in all treated plants and especially at the highest concentrations. At these concentrations the photosynthetic pathway was also altered as shown by the decrease of maximum quantum yield of photosystem II and the chlorophyll b content and by the chloroplast ultrastructural modifications. Starch storage was also investigated by microscopic observations. It was the highest at the high concentrations of the pollutant. The data suggested a correlation between starch storage and reduced growth; there was greater inhibition of plant growth than inhibition of photosynthesis, resulting in a surplus of carbohydrates that may be stored as starch. The investigation helps to understand the mechanism related to heavy metal tolerance of Lemna minor and supplies information about the behavior of this species widely used as a biomarker.

Cloning and expression analysis of a Petunia hybrida flower specific mitotic-like cyclin.

A cyclin cDNA clone (Pethy;CycB1;1) was isolated from a Petunia hybrida ovary specific cDNA library. Sequence comparison revealed that Pethy;CYCB1;1 protein is highly homologous to mitotic B1 cyclins. Northern analysis and in situ hybridisation experiments showed that its expression is developmentally regulated and restricted to flower organs. We have attempted to define some of the cell division patterns which contribute to shaping each floral organ by analysing Pethy;CycB1;1 expression on Petunia flower sections. While in sepals, epidermis and parenchyma cell division patterns were comparable, there were two distinct cell division patterns in petals. In the epidermis, Pethy;CYCB1;1 expression was found both at the petal tip and along epidermis, whereas in the parenchyma only at the petal tips. In reproductive organs cell divisions were detected only in sporophytic tissues. No signals were detected inside meiotic cells.

Technetium-99m-MIBI scintigraphy in the assessment of neoadjuvant chemotherapy in breast carcinoma.

UNLABELLED: Presurgical neoadjuvant chemotherapy (PSNC) is the treatment of choice for patients with locally advanced breast carcinoma (LABC). Accurate assessment of tumor response is important in planning subsequent treatments. Conventional response assessment by mammography and clinical evaluation is not fully reliable. This study evaluates the diagnostic yield of serial 99mTc-MIBI scintigraphy in the assessment of LABC response to PSNC. METHODS: Twenty-nine patients affected by LABC underwent clinical, mammographic and 99mTc-MIBI scintigraphy before and after 3 cycles of FEC (500 mg/m2 5-fluorouracil, 50 mg/m2 epirubicin and 400 mg/m2 cyclophosphamide) on Days 1 and 8. Surgery was planned for 15 days after the third cycle of chemotherapy. Pathological status was obtained after surgery in all patients. RESULTS: Sensitivities (i.e., true-positive ratios) for a correct prediction of tumor presence after PSNC were 65% for scintigraphy, 35% for clinical evaluation and 69% for mammography. Specificities (i.e., true-negative ratios) for a correct prediction of tumor absence after PSNC were 100% for scintigraphy, 67% for clinical evaluation and 33% for mammography. Technetium-99m-MIBI uptake in this series did not correlate with P-170 expression, proliferating cell nuclear antigen, Her-2/neu oncogene protein, antihuman endothelial cell CD31 antigen and estrogenic and progestinic receptor status. CONCLUSION: Technetium-99m-MIBI scintigraphy is effective in monitoring the response to PSNC in LABC patients. Its diagnostic yield is clearly superior to clinical evaluation alone. Scintigraphy performs as does mammography in patients with negative response, but it is clearly superior in patients with positive response.

Desmin-free cardiomyocytes and myocardial dysfunction in end stage heart failure.

Our aim was to evaluate the desmin content in the myocardial tissue of patients with end-stage heart failure of ischaemic origin and to assess its role on cardiac function. We studied 18 explanted hearts from patients transplanted for end-stage heart failure due to ischaemic cardiomyopathy (ICM). Control myocardial tissue was obtained from the cardiac biopsies of six women with breast cancer taken prior to commencing chemotherapy with anthracyclines, four male donors for heart transplantation and two autoptic hearts from patients who died due to non-cardiac events. Myocardial tissue, obtained from the left ventricle (remote zone from infarcted area), was analyzed by light and confocal immunochemistry (desmin) microscopy. The desmin content of myocardial tissue was obtained by real-time PCR. Cardiac function was evaluated by echocardiographic and right heart catheterization data, obtained before heart transplantation. Confocal microscopy evaluation showed a significant decrease in the number of desmin-positive myocytes (P<0.01) in ICM hearts compared to controls. At real-time PCR evaluation, there was a reduction (P<0.01) in desmin content in the ICM patients compared to controls. A negative correlation was found between desmin-free cardiomyocytes and ejection fraction (EF) (r=-0.834; P<0.02) on echocardiogram. A negative relationship (r=-0.688) was also found between desmin-negative myocytes and capillary wedge pressure. In conclusion, the myocardial tissue of patients with end-stage heart failure of ischaemic origin, shows a decreased number in desmin-positive myocytes at immunochemistry evaluation compared to normal individuals. This deficiency in cytoskeletal intermediate filament content is associated with reduced cardiac function.

[Colon volvulus]. / Volvolo del colon.

The authors report a clinical case and review the international literature. After an analysis of the incidence and the predisposing factors causing this disorder, they focus attention on the question of therapy. In forms with vital loop non-surgical derotation must be attempted. In the event of the failure of non-invasive treatment and to prevent recidivation these forms are treated surgically, also using videolaparoscopy. In forms with non-vital loop, surgery consists of the section of the necrotic segment and preparation of anastomosis which may be immediate or deferred depending on general and local conditions.

[Klippel-Trenaunay syndrome: clinical and surgical aspects]. / La sindrome di Klippel-Trenaunay. Aspetti clinici e chirurgici.

The Klippel-Trenaunay syndrome is a rare syndrome of uncertain etiology. The characteristic elements are flat angiomatosis, hypertrophy of soft tissue and bone tissue and alterations of the venous system, with the exclusion of hemodynamically significant arteriovenous fistulae. The authors report a clinical case and review the international literature. Treatment is conservative in the majority of cases; surgery is reserved for patients with disabling morphological and functional alterations.

[Biliary ileus. Case report and therapeutic considerations]. / Ileo biliare. Presentazione di un caso e considerazioni clinico-terapeutiche.

The authors, after having described, a case of biliary ileus, analyse the principal pathogenetic aspects of the disease, and underline the diagnostic and therapeutic difficulties. They believe that the simple enterolithotomy represents, initially, the best therapy, in particular with patients in poor clinical conditions.

[Diverticulitis of the cecum: description of 2 clinical cases]. / La diverticolite del cieco. Descrizione di due casi clinici.

The authors describe two cases of cecal diverticulitis pointing out the rarity of the disease in particular in Western countries. The disease is often asymptomatic although in the case of diverticulitis, which represent the most frequent complication of the disease, the patient becomes symptomatic. The symptomatology in these cases is similar to an acute appendicitis with which it is constantly confused. Surgery is the most common therapy and the type of operation is based on the clinical condition of the patient and local anatomical-surgical situation.

[Leiomyosarcoma of the mesentery. A case report and clinico-diagnostic considerations]. / Il leiomiosarcoma del mesentere. Descrizione di un caso e considerazioni clinico-diagnostiche.

Leiomyosarcoma of the mesentery is a rare neoplasm. The clinical manifestation is non-specific and preoperative diagnosis is often late. Mitotic activity shows no significant correlation with the biological behavior of neoplasm. The surgical excision is the therapy of choice but local recurrence is high. The authors report a case of leiomyosarcoma which was treated by surgical removal and underline the diagnostic difficulties.