ALTERAZIONI ISTOPATOLOGICHE INDOTTE DA CELLULE UMANE LAM/TSC IN TOPI NUDI: UN MODELLO DI LINFANGIOLEIOMIOMATOSI. REVERSIONE DEL DANNO CON ANTICORPO ANTI-EGFR

LAM is a rare and progressive disease characterized by widespread proliferation of abnormal smooth muscle-like cells (LAM cells). LAM cells cause cystic destruction of lung parenchyma, abdominal tumours (angiomyolipoma, AML) and infiltration of axial lymphatics in torax and abdomen (adenopathy and lymphangioleiomyoma). LAM occurs sporadically or in association with tuberous sclerosis complex (TSC), an inherited disorder with variable penetrance, which results from mutations in TSC1 or TSC2 genes. TSC1 or TSC2 genes, encoding hamartin and tuberin respectively, regulate mammalian target of rapamycin (mTOR). LAM affects primarly women of child-bearing age and the mechanisms causing the disease are not yet clarified. To explain the multisystemic clinical manifestations of LAM an experimental model is needed to study the pathological mechanism causing LAM. It may help to explain how LAM cells migrate from tissue to tissue and to develop a pharmacological approach. We recently isolated and characterized \u3b1-actin positive smooth muscle cells from chylous of a patient affected by LAM/TSC (LAM/TSC cells). These circulating cells showed reactivity to HMB45 and CD44v6 antibodies, markers of TSC and LAM, and bear a germline TSC2 mutation in exon 21. Like TSC2 smooth muscle cells previously isolated (TSC2-/- and TSC2-/meth ASM cells), LAM/TSC cells from chylous required epidermal growth factor (EGF) to proliferate and the blockade of EGF receptor (EGFR) caused progressive cell death. To better study LAM pathogenesis we developed a procedure for a quick invasion of the respiratory system by endonasally administrating LAM/TSC cells. LAM/TSC cells were administrated in immunodeficient female nude mice (nu/nu Hsd: athymic nude mice, 3 weeks old) and after 26 weeks anti-EGFR antibody and rapamycin were intraperitoneally injected 2 times a week for 4 weeks. 30 weeks after endonasal administration LAM/TSC cells were detected in lungs, lymph nodes and uterus. In lung parenchyma, LAM/TSC cells proliferated and caused cystic destruction with emphysematous-like picture such as in LAM patient lungs. This lesion and the proliferating rate were reverted by anti-EGFR antibody, while rapamycin was less effective and caused hemoptysis. In lungs blood vessel number was increased and, using LYVE-1 antibody, a significant increase of lymphatic vessel density (LVD) was observed in animals that received LAM/TSC cells suggesting a possible correlation between LAM/TSC cells and lymphangiogenesis. LVD decreased following anti-EGFR antibody and rapamycin treatments. When treatments were stopped, hemoptysis caused by rapamycin was reverted. Anti-EGFR antibody was more effective than rapamycin in reducing lung injury caused by LAM/TSC cells administration and in decreasing lymphangiogenesis. In some lung parenchyma noduli were detected. Such in lungs of LAM patients they were positive to human COX IV, ER, PR and phosphoS6. In lymph nodes, LAM/TSC cells promoted a lymphatic vessel invasion, as showed by PROX-1-reactivity. Anti-EGFR antibody and rapamycin treatments decreased lymphatic vessels. Differently from lungs, blood vessels in lymph nodes were not altered after LAM/TSC cells administration, as shown by CD31 immunoreactivity. LAM/TSC cells were also detected in uteri where they promoted a significant increase of cells with high levels of estrogen (ER) and progesterone receptors (PR). In uteri afterLAM/TSC cells administration the morphological structure was unchanged. Pharmacological treatments decreased ER and PR expressions. Our data show that endonasal administration of cells isolated from chylous of LAM/TSC patient developed a mouse LAM model. LAM/TSC cells invaded lungs, lymph nodes and uteri causing LAM-like lesions. Anti-EGFR antibody is more effective than rapamycin in promoting lung restauration and reducing lymphangiogenesis; its efficacy persists also when treatment is stopped. These data suggest that anti-EGFR antibody treatment may represent a useful pharmacological approach for LAM therapy

Cloud Chamber: A Performance with Real Time Two-Way Interaction between Subatomic Particles and Violinist

‘Cloud Chamber’ - a composition by Alexis Kirke, Antonino Chiaramonte, and Anna Troisi - is a live performance in which the invisible quantum world becomes visible as a violinist and subatomic particle tracks interact together. An electronic instrument was developed which can be “played” live by radioactive atomic particles. Electronic circuitry was developed enabling a violin to create a physical force field that directly affects the ions generated by cosmic radiation particles. This enabled the violinist and the ions to influence each other musically in real time. A glass cloud chamber was used onstage to make radioactivity visible in bright white tracks moving within, with the tracks projected onto a large screen

The echocardiography diagnosis of cor pulmonale in a horse

The authors describe the two-dimensional and Doppler signs that characterize the pulmonary hypertension caused by chronic obstructive pulmonary disease (COPD) in a horse

Diagnostic factors for recurrent pregnancy loss: an expanded workup

Purpose: There is limited information on the risk factors for recurrent pregnancy loss (RPL). Methods: In this study, a patient-based approach was used to investigate the possible involvement and relative relevance of a large number of diagnostic factors in 843 women with RPL who underwent an extensive diagnostic workup including 44 diagnostic factors divided into 7 major categories. Results: The rates of abnormalities found were: (1) genital infections: 11.74%; (2) uterine anatomic defects: 23.72%; (3) endocrine disorders: 29.42%; (4) thrombophilias: 62%; (5) autoimmune abnormalities: 39.2%; (6) parental karyotype abnormalities 2.25%; (7) clinical factors: 87.78%. Six hundred and fifty-nine out of eight hundred and forty-three women (78.17%) had more than one abnormality. The mean number of pregnancy losses increased by increasing the number of the abnormalities found (r = 0.86949, P < 0.02). The factors associated with the highest mean number of pregnancy losses were cervical isthmic incompetence, anti-beta-2-glycoprotein-1 antibodies, unicornuate uterus, anti-prothrombin A antibodies, protein C deficiency, and lupus anticoagulant. The majority of the considered abnormalities had similar, non-significant prevalence between women with 2 versus ≥ 3 pregnancy losses with the exception of age ≥ 35 years and MTHFR A1298C heterozygote mutation. No difference was found between women with primary and secondary RPL stratified according to the number of abnormalities detected (Chi-square: 8.55, P = 0.07). In these women, the only factors found to be present with statistically different rates were age ≥ 35 years, cigarette smoking, and genital infection by Ureaplasma. Conclusion: A patient-based diagnostic approach in women with RPL could be clinically useful and could represent a basis for future research

Early intestinal perforation secondary to congenital mesenteric defects

Gastrointestinal perforation (GIP) in preterm neonates may be idiopathic, due to necrotizing enterocolitis (NEC), or mechanical obstruction. The predominant cause of GIP in the neonatal period is NEC. Differential diagnosis with congenital malformations, including mesenteric defects leading to internal hernias, is mandatory if the onset is early. We describe two newborns with trans-mesenteric herniation resulting in GIP, and we discuss the presence of possible additional risk factors such as prematurity and predisposing vascular disruption in connective tissue disorders (Ehlers-Danlos syndrome), twinning, and use of assisted reproductive technologies. These cases prompted us to review our exploratory laparotomies performed for intestinal obstruction, complicated/or not with perforation, to identify the frequency of neonatal trans-mesenteric hernias in a referral hospital. The prevalence of GIP and of internal hernia was 25% and 3.3%, respectively. In conclusion, time-onset and particular conditions associated with GIP should lead to a high index of suspicion for internal hernias in order to achieve appropriate diagnosis and therapy

P38 SNP PROFILING OF KIDNEY TRANSPLANT PATIENTS: HIGHLIGHTING THE DIFFERENCES IN THE GENETICAL BACKGROUND TO MAXIMALIZE IMMUNOSUPPRESSIVE THERAPIES

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Amblyraja georgiana, Antarctic Starry Skate

The Antarctic Starry Skate (Amblyraja georgiana) is a medium-sized (to 115 cm total length) deepwater skate that occurs in the Southeast Pacific Ocean off southern Chile, in the Southwest Atlantic Ocean off southern Argentina and the Falkland Islands (Malvinas), in the Atlantic and Pacific Antarctic Oceans from South Georgia Island and the Antarctic Peninsula to the Ross Sea, and in the Indian Antarctic off the Crozet Islands. It is demersal on continental and insular slopes at depths of 20?1,255 m, and is captured as bycatch in trawl and longline fisheries, particularly those targeting Patagonian Toothfish (Dissostichus eleginoides). There are no population size estimates for this skate, and it is not clear what the current population trend is. Although estimates of bycatch around South Georgia and the Ross Sea are comprised of a low percentage of overall estimated stock biomass, the demographic consequences are unknown and require further research. Furthermore, catch levels in other areas are unknown, and some specimens previously referred to as this species may include cryptic individuals of a yet-to-be-described species. Overall, it is not clear what level of fishing mortality this species is exposed to across its range, and further research is needed on distribution, population size and trend, and threats. Therefore, the Antarctic Starry Skate is assessed as Data Deficient.Fil: Pollom, R.. University Fraser Simon; CanadáFil: Acuña, E.. Universidad Católica del Norte; ChileFil: Bustamante, C.. Universidad de Antofagasta; ChileFil: Chiaramonte, Gustavo Enrique. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia". Estación Hidrobiológica de Puerto Quequén (sede Quequén); ArgentinaFil: Cuevas, J.M.. Wildlife Conservation Society; Estados UnidosFil: Herman, K.. Georgia Aquarium; Estados UnidosFil: Pompert, J.. No especifíca;Fil: Velez Zuazo, X.. No especifíca

LAM/TSC cell migration to uterus in an experimental model of lymphangioleiomyomatosis. Regulation by anti-epidermal growth factor receptor antibody and rapamycin

Lymphangioleiomyomatosis (LAM) is a rare lung disease affecting almost exclusively women, characterized by the invasion and abnormal proliferation of smooth muscle-like cells in pulmonary parenchyma and axial lymphatics. LAM cells bear mutations in Tuberous Sclerosis Complex (TSC) genes. It has been hypothesized that uterus might be the primary site of origin and one of the most frequent metastatic or disseminated site of LAM cells. We developed a mouse model to study the migratory and invasive properties of human LAM/TSC cells to the uterus. We also examined the action of rapamycin and anti-Epidermal Growth Factor Receptor (EGFR) antibody. LAM/TSC cells were endonasally administered to 3 week old immunodeficient female nude mice. 5 months later mice were divided in 4 groups: control, LAM/TSC cell-administered mice, LAM/TSC cell-administered mice treated with rapamycin, and LAM/TSC cell-administered mice treated with anti-EGFR antibody. Drugs were administered for one months. Uteri were analysed for the presence of human LAM/TSC cells by COX IV antibody, lymphangiogenesis by LYVE 1 expression and angiogenesis by counting blood vessels. LAM/TSC cells migrated to the uterus without causing any morphological lesion. Interestingly, LAM/TSC cells increased the number of blood vessels while did not cause any alteration in lymphatics vessels. Anti-EGFR antibody and rapamycin reduced the number of human LAM/TSC and counteracted the proliferation of blood vessels in uteri. Although both drugs did not change the expression of LYVE 1, localization of lymphatics was mainly in the perimetrium after drug treatment. Our data describe the strong invasive capability of human LAM/TSC cells which migrated to the uterus. LAM/ TSC cells presence is accompanied by increased angiogenesis. Anti-EGFR antibody and rapamycin were effective in reducing the LAM/TSC cell number and blood vessel proliferation

Anti-EGFR antibody reduces lung nodules by inhibition of EGFR-pathway in a model of lymphangioleiomyomatosis

EGFR belongs to the HER/Erb family of tyrosine kinase receptors and its activation in cancer cells has been linked with increased proliferation, angiogenesis, and metastasis. Lymphangioleiomyomatosis (LAM) is a rare, low-grade neoplasm that occurs sporadically or in association with tuberous sclerosis complex (TSC), a genetic, multisystem disorder characterized by hamartomas in several organs. From chylous of a LAM/TSC patient, we previously isolated smooth muscle-like LAM/TSC cells which proliferation depends on EGF and monoclonal antibodies anti-EGFR reduced proliferation and caused cell death. We demonstrated that the dependence from EGF was caused by the absence of tuberin. To study the role of EGFR pathway in vivo, we developed a mouse model by administration of LAM/TSC cells to female nude mice. LAM/TSC cells caused pulmonary airspace enlargement and, after 30 weeks, nodule formation which express EGFR. Anti-EGFR antibody decreased the number and dimension of lung nodules likely for the inhibition of Erk and S6 signaling, reversed the pulmonary alterations and reduced lymphatic and blood vessels. Moreover, in pulmonary nodules anti-EGFR antibody reduced the positivity to estrogen and progesterone receptors which enhance survival of LAM cells and Snail expression. These results suggest that the inhibition of EGFR signalling have a potential in treatment of LAM/TSC lung alterations