Mesenteric cyst and recurrent abdominal pain in a patient with Gorlin-Goltz syndrome: a case report.

Gorlin-Goltz syndrome (GGS) is an infrequent autosomal do-minant multisystemic disease with complete penetrance and variable expressivity. It is estimated to have an incidence of 1:50,000 - 1:150,000 cases with a M/F = 1:1. This report describes a case of recurrent abdominal pain due to a large mesenteric cyst in a 38-year-old female patient affected by a rare disease: Gorlin-Goltz syndrome.

Predictive factors of difficult procedure in octogenarians undergoing elective laparoscopic cholecystectomy: a single center experience.

AIM: To assess the feasibility and safety of laparoscopic cholecystectomy (LC) in very elderly patients with particular attention to the predicitive factors of difficulty. PATIENTS AND METHODS: All patients aged ≥ 80 undergoing elective LC for lithiasis at our institution since 1st January 2015 to 31st December 2015 were included in the study. Exclusion criteria were: a) acute cholecystitis; b) biliary pancreatitis; c) biliary tract neoplasms; d) urgent procedure. Pre-, intra- and postoperative data were recorded. RESULTS: During the study period, we performed 72 LC and we enrolled 17 patients aged ≥ 80 with a M:F = 5:12. Of these, 10 patients had a "difficult" cholecystectomy. In seven cases an intraoperative cholangiography (IOC) was performed. Postoperative course was regular but in two patients we had an Oddian spasm in 1st postoperative day. Female sex (p=0.03) and preoperative high level of serum amylase (p= 0.02) were significantly associated to difficult cholecystectomy in elderly patients. CONCLUSION: LC in octogenarians is feasible and safe. However, sex and serum amylase can help the surgeon to predict a more difficult procedure in elective LC. In this group of patients an approach based on the individual risk is desirable and the patient could be referred to a multidisciplinary approach.

A new radioguided procedure for localization and surgical treatment of neck node metastasis of papillary thyroid cancer.

AIM OF THE STUDY: We intended to use a radioguided technique for pre-operative localization of neck node recurrences in patients with papillary thyroid cancer (PTC) already submitted to thyroidectomy and radioiodine treatment. PATIENTS AND METHODS: We selected 20 patients affected by PTC with evidence of neck nodes recurrences at ultrasound examination. Our method has been derived from the Radioguided Occult Lesion Localization technique used for pre-operative localization of occult breast lesions. The technique involves the inoculation of human albumin macroaggregates labeled with radioactive technetium (0.4 mCi in a volume of 0.05 ml) directly in the suspicious lesion, under ultrasound guidance. The persistence of the radioactive tracer in the nodes is confirmed by a scintigraphy performed 2 h after injection. During surgery, a gamma detecting probe is used to locate the suspicious lesions as "hot spots". RESULTS: Fifty lymph-nodes were injected with the tracer. All radiolabeled lymph-nodes were located and removed during surgery. At histology, metastasis of PTC was confirmed in 38/50 (76%) lymph-nodes. At least one metastatic lymph-node per patient was removed. In 8/20 (40%) patients, reactive lymphoid hyperplasia was found in 12/50 (24%) lymph-nodes. CONCLUSIONS: This radioguided technique has been highly effective for localization and surgical treatment of suspicious lymph-node detected at neck ultrasound and may play a valuable role in case of node metastases of thyroid cancer that show no radioiodine uptake.

[Cryptococcosis in AIDS patients: case study from 1996 to 2006 in Paroissien Hospital]. / Criptococosis en pacientes con SIDA: estudio de casos en el Hospital Paroissien en el período 1996-2007.

Clinic, epidemiological and microbiological characteristics of 128 episodes of cryptococcosis were retrospectively evaluated in 106 positive HIV patients hospitalized at the Paroissien Hospital on period 1996-2007. There were 75 male and 31 female patients, with a median age of 34 years, ranging from 20 to 68 years. Addiction to intravenous drugs was the main cause of HIV infection in 55 patients (51.9%). Cryptococcosis was detected as single episode in 85 patients (80.2%) and as relapse in 19 (17.9%). It appeared as the first marking disease in 36 patients (34.0%). Diagnosis was established in 116 episodes by CSF study (Indian ink, culture, antigen detection), in 9 cases by Cryptococcus sp. recovery from blood cultures, and in 3 cases by antigen detection in patient's serum with a latex reactive. Neurological symptomatology, with headache and fever as the most common signs, was presented by 89.6% of patients. Induction treatment was done in all cases with amphotericin B, and maintenance treatment was carried out with fluconazole. Mortality rate was 35.8%, being higher in those patients who had suffered relapses (41.3%) compared to those who presented a first episode of the mycosis (33.3%).

Saporin as a novel suicide gene in anticancer gene therapy.

We used a non-viral gene delivery approach to explore the potential of the plant saporin (SAP) gene as an alternative to the currently employed suicide genes in cancer therapy. Plasmids expressing cytosolic SAP were generated by placing the region encoding the mature plant ribosome-inactivating protein under the control of cytomegalovirus (CMV) or simian virus 40 (SV40) promoters. Their ability to inhibit protein synthesis was first tested in cultured tumor cells co-transfected with a luciferase reporter gene. In particular, SAP expression driven by CMV promoter (pCI-SAP) demonstrated that only 10 ng of plasmid per 1.6 x 10(4) B16 cells drastically reduced luciferase activity to 18% of that in control cells. Direct intratumoral injection of pCI-SAP complexed with either lipofectamine or N-(2,3-dioleoyloxy-1-propyl) trimethylammonium methyl sulfate (DOTAP) in B16 melanoma-bearing mice resulted in a noteworthy attenuation of tumor growth. This antitumor effect was increased in mice that received repeated intratumoral injections. A SAP catalytic inactive mutant (SAP-KQ) failed to exert any antitumor effect demonstrating that this was specifically owing to the SAP N-glycosidase activity. Our overall data strongly suggest that the gene encoding SAP, owing to its rapid and effective action and its independence from the proliferative state of target cells might become a suitable candidate suicide gene for oncologic applications.

[Ultrasound-guided vacuum-assisted biopsy: review of 382 cases]. / Macrobiopsies assistées par le vide sous guidage échographique: à propos de 382 cas.

PURPOSE: To evaluate the use of US-guided vacuum biopsy for diagnosis and treatment of probably benign breast masses. MATERIALS AND METHOD: Retrospective review of 382 US guided vacuum biopsies over a 44 months period (september 2001 to may 2005) with the 11-g handheld mammotome. A total of 308 benign tumors, 59 borderline lesions and 15 carcinomas were diagnosed. The average number of specimens is 13.1 (3-37). Surgical resection has been systematic for carcinomas and selective for papillomas. Surgical correlation (n:35) or mammographic follow-up (n:347) are presented. RESULTS: Complete removal occurred in 371/382 (97.1%) immediately after biopsy and 337/382 (88.2%) after one month: 138/142 (93.7%) for fibroadenomas and 52/53 (98.1%) papillomas less than 15 mm. Open surgical biopsy was carried out for 35 patients on the basis of incomplete removal (3 cases) or histologic findings (8 invasive carcinomas, 7 ductal carcinoma in situ, 3 atypical ductal hyperplasia, 1 fibrocystic changes with atypia and 11 papillomas). No lesion was under-diagnosed and the rate of avoided surgery was 94.5%. Of the 347 lesions that were not surgically biopsied (42 borderline lesions and 305 benign lesions), 337 were monitored at 1-43 months (average: 20 months, > or =24 months: 57 patients). Ten underwent additional biopsy but no missed cancer was detected. Patients tolerance was good or very good in 83%, and the complication rate was 1.3%. CONCLUSION: US-guided vacuum biopsy is an accurate and well tolerated technique. It is an alternative to surgery for masses less than 15 mm including fibroadenomas and papillomas or in patients with imaging-histologic discordance at core biopsy.

Abdominal radiography findings in small bowel anastomoses with V-BAR.

PURPOSE: The aim of the present paper is to describe the normal radiological findings and the fragmentation time of the VALTRAC BAR (V-BAR). MATERIALS AND METHODS: Ten cases of Roux-en-Y jejuno-jejunostomy with anastomosis with a n.25 Valtrac ring for gastric cancer were followed up during the postoperative period by plain abdominal X-ray examination. RESULTS: Abdominal X-ray examination showed V-BAR fragmentation between day 14 and 21 after surgery. In one case only did fragmentation fail and the V-BAR migrated resulting in mechanical ileus. CONCLUSIONS: The possible non-fragmentation and migration of the V-BAR confirms the need to follow up patients with plain abdominal radiography performed at short intervals during the postoperative period.

Gastric cancer in the elderly patient.

The incidence of gastric cancer is increasing in elderly patients, unlike what it's being observed in the younger patients. The aim of this study is to evaluate the impact of the age on the resecability, resection type (R0-R1-2), morbidity, mortality and survival rate. The higher mortality observed in the elderly patients, the discussed role of an extended lymphadenectomy and the poor survival rate, would lead to a surgical approach which mostly tends to the palliation of the symptoms than the obtainment of a curative resection. But, on the other hand, the impossibility of making chemotherapy in most of the over 75 year old patients, lead us to a R0 surgery treatment.

[Total gastrectomy for gastric cancer: can the type of lymphadenectomy condition the long-term results?]. / Gastrectomia totale per cancro gastrico: può il tipo di linfadenectomia condizionare i risultati a distanza?

INTRODUCTION: Gastric cancer is the second tumor for frequency in the world. Surgery is still the only curative treatment. Good results in terms of long distance survival, postoperative morbidity and mortality have been achieved in the last years. The extension of lymphadenectomy is an important and discussed matter and it is not clear if lymphadenectomy may contribute to improve the surgical results. The Japanese surgeons were the first ones, in the 60's, to introduce a D2-D3 extended lymphadenectomy, but the real benefits of this technique are still being discussed. Indeed lymphonodal metastasis seem to be one of the most important prognostic factors in the gastric cancer and the level and the number of metastatic nodes are useful to predict the patients' survival. The aim of this study is to value the D2 lymphadenectomy in the patients who were treated with total gastrectomy for gastric adenocarcinoma, comparing the results both with the D1 lymphadenectomy and the D3-D4, paying attention to the survival rates related with the lymphonodal dissection. PATIENTS AND METHOD: From 1998 to 2004, we studied 87 patients with gastric cancer. Out of 78 patients treated surgically, 9 were judged unresectable. Out of 69 patients treated surgically, one died before surgery and so he was put away by this study. All the patients were treated with total gastrectomy and a GI tract reconstruction by Roux's Y termino-lateral esophageal-jejunal anastomosis. In 20 patients we also made a splenectomy. We followed the Japanese Research Society for Gastric Cancer guidelines, according to which nodes are gathered into 16 levels and divided in 4 groups (N1-N4) depending on the cancer localization. The extension of the lymphadenectomy has been classified according to the level of the removed nods. The patients were divided into 3 groups. First group: patients undergone a total gastrectomy with D1 lymphadenectomy. Second group: patients undergone D2 lymphadenectomy. Third group: patients undergone D3 and D4 lymphadenectomy. The data we obtained let us value the survival rate. RESULTS: Out of the 78 patients treated, 69 were resected with a 88.5% resection rate (69/78). Perioperatory mortality was 1.4% (1/69). Global survival was 53.8% (44/68). The 5 years survival for the Ia stage was 82.6%, 89.3% for the Ib stage, 67.8% for the II stage, 56.6% for the IIIa, 16.8% for the IIIb and 0% for the IV stage. In addition the 5 years survival in the patients without lymphnodal metastasis was 79%, much higher than the 30.6% obtained in the patients with lymphonodal metastasis (p <0.0001). In the patients who underwent D1 lymphadenectomy, survival was 73.4%, while we obtained a result of 70.4% and 13.8% respectively in the D2 and D3-D4 (p <0.05). In the advanced stages (IIIa, IIIb, IV), the survival rate in the patients with lymphadenectomy D2 vs D1 vs D3-D4 was 38.9% vs 0% vs 36.3% (p <0.0001). The survival rate based on the extension of the lymphadenectomy in the patients with lymphonodal metastasis resulted to be much higher in those patients who underwent a D2 lymphadenectomy (43.5% in 5 years) compared both to D1 (0% in 6 months) and (29.5% in 5 years) D3-D4 lymphadenectomy. CONCLUSIONS: The relation between long distance survival and extension of the lymphadenectomy in the patients with gastric adenocarcinoma is still being discussed. Different studies show the importance of a careful lymphadenectomy as the main mean for a better long distance survival in the patients with gastric cancer. Other studies showed a link between gastric cancer prognosis and number of positive nodes. If more than 7-8 nodes are affected with metastasis, prognosis is usually poor. Therefore the presence of nod metastasis has a negative influence in the prognosis of this tumor; for this reason D2 lymphadenectomy is the technique to be used for the treatment of the gastric adenocarcinoma, both for a good staging and a better long distance survival, but particularly in the advanced cancers which are, at the moment, the highest number of cases that reach the surgeon's attention.

[Antithrombotic prophylaxis in day surgery: proposal of a clinical protocol]. / La profilassi antitrombotica in day surgery. Proposta di un protocollo clinico.

The considerable diffusion of the day surgery has new problems such as the use of antithrombotic prophylaxis, now carried out systematically in major surgery. An the basis of the poor data present in the literature, a protocol of study is proposed to evaluate the real effectiveness of this type of prophylaxis in DS, dividing the patients into 3 groups, according to the patient's risk factors to the intervention and to the disease.

Biochemical characterization of bona fide polycystin-1 in vitro and in vivo.

The most common form of autosomal dominant polycystic kidney disease (PKD) results from mutation of the PKD1 gene on chromosome 16p13.3. The gene encodes a 14-kb messenger RNA that is predicted to express a 462-kd membrane protein. The gene product, polycystin-1, has a large extracellular portion composed of a novel combination of protein-protein interacting domains and is postulated to be a plasma membrane receptor involved in cell-cell/matrix interactions. However, slow progress has been made in the characterization of polycystin-1 or the determination of its function. In fact, the protein is expressed at very low levels in tissues and cell lines and previous efforts directed at expression of recombinant protein had been largely unsuccessful. We have recently developed constructs of full-length human PKD1 complementary (cDNA) that can be expressed in both a stable and transient fashion in mammalian cells. We used these systems to characterize our antibodies and to track the protein in vivo. We report here the first biochemical characterization of recombinant polycystin-1 and show that the protein is a 520-kd glycosylated polypeptide with an unglycosylated core of 460 kd. Subcellular fractionation as well as biotinylation studies confirmed that the protein is plasma-membrane associated. Furthermore, we show that the recombinant protein localizes to cell-cell junctions in polarized madin darby canine kidney cells as revealed by indirect immunofluorescence. Our data represent the first characterization of polycystin-1 performed under highly controlled conditions.

Altered behavioral rhythms and clock gene expression in mice with a targeted mutation in the Period1 gene.

A group of specialized genes has been defined to govern the molecular mechanisms controlling the circadian clock in mammals. Their expression and the interactions among their products dictate circadian rhythmicity. Three genes homologous to Drosophila period exist in the mouse and are thought to be major players in the biological clock. Here we present the generation of mice in which the founding member of the family, Per1, has been inactivated by homologous recombination. These mice present rhythmicity in locomotor activity, but with a period almost 1 h shorter than wild-type littermates. Moreover, the expression of clock genes in peripheral tissues appears to be delayed in Per1 mutant animals. Importantly, light-induced phase shifting appears conserved. The oscillatory expression of clock genes and the induction of immediate-early genes in response to light in the master clock structure, the suprachiasmatic nucleus, are unaffected. Altogether, these data demonstrate that Per1 plays a distinct role within the Per family, as it may be involved predominantly in peripheral clocks and/or in the output pathways of the circadian clock.

[AIDS-associated meningeal cryptococcosis in the Hospital Diego Paroissien from 1996-1999]. / Criptococosis meníngea asociada al SIDA en el Hospital Diego Paroissien en el período 1996/1999.

A total of 148 patients with a diagnosis of HIV infection were studied in order to evaluate the incidence of meningeal cryptococcosis, including epidemic, immunologic and diagnostic characteristics. The diagnosis of cryptococcosis was carried out by direct examination with India ink and culture in Sabouraud agar of CSF in 28 patients (93.3%) and by blood cultures (lysis-centrifugation) in 2 patients (6.6%). All the isolated strains were identified as Cryptococcus neoformans. The incidence was 20.3% (30 patients). The preponderant risk behavior was endovenous drug addiction, and it was observed in 18 patients (60%). The symptomatology that prevailed was headache (87%). The median age was 28 years. At diagnosis, the immunologic impairment was severe (CD4+ lymphocyte count < 200) in 90% of patients. We found that 86.7% of patients had not completed their primary studies and only 13.3% had completed secondary studies. Although the acute mortality was high (36.7%), it was observed that all the patients who survived (24%) had been treated with anti-retroviral drugs.

Criptococosis meníngea asociada al SIDA en el Hospital Diego Paroissien en el período 1996/1999 / AIDS-associated meningeal cryptococcosis in the Hospital Diego Paroissien from 1996-1999

A total of 148 patients with a diagnosis of HIV infection were studied in order to evaluate the incidence of meningeal cryptococcosis, including epidemic, immunologic and diagnostic characteristics. The diagnosis of cryptococcosis was carried out by direct examination with India ink and culture in Sabouraud agar of CSF in 28 patients (93.3) and by blood cultures (lysis-centrifugation) in 2 patients (6.6). All the isolated strains were identified as Cryptococcus neoformans. The incidence was 20.3 (30 patients). The preponderant risk behavior was endovenous drug addiction, and it was observed in 18 patients (60). The symptomatology that prevailed was headache (87). The median age was 28 years. At diagnosis, the immunologic impairment was severe (CD4+ lymphocyte count < 200) in 90 of patients. We found that 86.7 of patients had not completed their primary studies and only 13.3 had completed secondary studies. Although the acute mortality was high (36.7), it was observed that all the patients who survived (24) had been treated with anti-retroviral drugs.

Polycystin-1, the gene product of PKD1, induces resistance to apoptosis and spontaneous tubulogenesis in MDCK cells.

The major form of autosomal dominant polycystic kidney disease (ADPKD) results from mutation of a gene (PKD1) of unknown function that is essential for the later stages of renal tubular differentiation. In this report, we describe a novel cell culture system for studying how PKD1 regulates this process. We show that expression of human PKD1 in MDCK cells slows their growth and protects them from programmed cell death. MDCK cells expressing PKD1 also spontaneously form branching tubules while control cells form simple cysts. Increased cell proliferation and apoptosis have been implicated in the pathogenesis of cystic diseases. Our study suggests that PKD1 may function to regulate both pathways, allowing cells to enter a differentiation pathway that results in tubule formation.

Glomerular filtration is required for transfection of proximal tubular cells in the rat kidney following injection of DNA complexes into the renal artery.

Gene transfer to the kidney can be achieved with various DNA vectors, resulting in transgene expression in glomerular or tubular districts. Controlling transgene destination is desirable for targeting defined renal cells for specific therapeutic purposes. We previously showed that injection of polyplexes into the rat renal artery resulted in transfection of proximal tubular cells. To investigate whether this process involves glomerular filtration of the DNA-containing particles, fluorescent polyethylenimine polyplexes were prepared, containing fluoresceinated poly-L-lysine. This allowed visualization of the route of the particles into the kidney. Our polyplexes were filtered through the glomerulus, since fluorescent proximal tubuli were observed. Conversely, fluorescent lipopolyplexes containing the cationic lipid DOTAP were never observed in tubular cells. Size measurements by laser light scattering showed that the mean diameter of polyplexes (93 nm) was smaller than that of lipopolyplexes (160 nm). The size of the transfecting particles is therefore a key parameter in this process, as expected by the constraints imposed by the glomerular filtration barrier. This information is relevant, in view of modulating the physico-chemical properties of DNA complexes for optimal transgene expression in tubular cells. Gene Therapy (2000) 7, 279-285.

Development and characterization of pituitary GH3 cell clones stably transfected with a human proinsulin cDNA.

Successful beta-cell replacement therapy in insulin-dependent (type I) diabetes is hindered by the scarcity of human donor tissue and by the recurrence of autoimmune destruction of transplanted beta cells. Availability of non-beta cells, capable of releasing insulin and escaping autoimmune recognition, would therefore be important for diabetes cell therapy. We developed rat pituitary GH3 cells stably transfected with a furin-cleavable human proinsulin cDNA linked to the rat PRL promoter. Two clones (InsGH3/clone 1 and 7) were characterized in vitro with regard to basal and stimulated insulin release and proinsulin transgene expression. Mature insulin secretion was obtained in both clones, accounting for about 40% of total released (pro)insulin-like products. Immunocytochemistry of InsGH3 cells showed a cytoplasmic granular insulin staining that colocalized with secretogranin II (SGII) immunoreactivity. InsGH3 cells/clone 7 contained and released in vitro significantly more insulin than clone 1. Secretagogue-stimulated insulin secretion was observed in both InsGH3 clones either under static or dynamic conditions, indicating that insulin was targeted also to the regulated secretory pathway. Proinsulin mRNA levels were elevated in InsGH3 cells, being significantly higher than in betaTC3 cells. Moreover, proinsulin gene expression increased in response to various stimuli, thereby showing the regulation of the transfected gene at the transcriptional level. In conclusion, these data point to InsGH3 cells as a potential beta-cell surrogate even though additional engineering is required to instruct them to release insulin in response to physiologic stimulations.

Cross-talk in signal transduction: Ras-dependent induction of cAMP-responsive transcriptional repressor ICER by nerve growth factor.

The CREM gene encodes both activators and repressors of cAMP-induced transcription. By virtue of an alternative, intronic promoter within the gene, the ICER (Inducible cAMP Early Repressor) isoform is generated. ICER acts as a dominant negative regulator and is cAMP-inducible in various neuroendocrine cells and tissues. ICER negatively autoregulates its own expression, and appears to participate in the molecular events governing oscillatory hormonal regulations. Here we report that ICER is inducible with nerve growth factor (NGF). This is the first example of cAMP-independent induction of ICER expression. Importantly, induction by NGF occurs via a subset of the CREs present in the ICER promoter which were previously shown to direct cAMP-inducibility. ICER induction correlates with a NGF-mediated phosphorylation of CREB. Both CREB phosphorylation and ICER inducibility require an intact Ras-dependent signalling pathway. We show that increased ICER levels result in the attenuation of c-fos expression. The activation of a powerful repressor of cAMP-responsive transcription by NGF, whose transduction signalling is cAMP-independent, constitutes a notable example of nuclear cross-talk and thus is likely to have relevant physiological implications.