Induction of Cytoplasmic Rods and Rings Structures by Inhibition of the CTP and GTP Synthetic Pathway in Mammalian Cells

Background: Cytoplasmic filamentous rods and rings (RR) structures were identified using human autoantibodies as probes. In the present study, the formation of these conserved structures in mammalian cells and functions linked to these structures were examined. Methodology/Principal Findings: Distinct cytoplasmic rods (,3–10 mm in length) and rings (,2–5 mm in diameter) in HEp-2 cells were initially observed in immunofluorescence using human autoantibodies. Co-localization studies revealed that, although RR had filament-like features, they were not enriched in actin, tubulin, or vimentin, and not associated with centrosomes or other known cytoplasmic structures. Further independent studies revealed that two key enzymes in the nucleotide synthetic pathway cytidine triphosphate synthase 1 (CTPS1) and inosine monophosphate dehydrogenase 2 (IMPDH2) were highly enriched in RR. CTPS1 enzyme inhibitors 6-diazo-5-oxo-L-norleucine and Acivicin as well as the IMPDH2 inhibitor Ribavirin exhibited dose-dependent induction of RR in.95 % of cells in all cancer cell lines tested as well as mouse primary cells. RR formation by lower concentration of Ribavirin was enhanced in IMPDH2-knockdown HeLa cells whereas it was inhibited in GFP-IMPDH2 overexpressed HeLa cells. Interestingly, RR were detected readily in untreated mouse embryonic stem cells (.95%); upon retinoic acid differentiation, RR disassembled in these cells but reformed when treated with Acivicin

Identification of an autoantibody panel to separate lung cancer from smokers and nonsmokers

<p>Abstract</p> <p>Background</p> <p>Sera from lung cancer patients contain autoantibodies that react with tumor associated antigens (TAAs) that reflect genetic over-expression, mutation, or other anomalies of cell cycle, growth, signaling, and metabolism pathways.</p> <p>Methods</p> <p>We performed immunoassays to detect autoantibodies to ten tumor associated antigens (TAAs) selected on the basis of previous studies showing that they had preferential specificity for certain cancers. Sera examined were from lung cancer patients (22); smokers with ground-glass opacities (GGOs) (46), benign solid nodules (55), or normal CTs (35); and normal non-smokers (36). Logistic regression models based on the antibody biomarker levels among the high risk and lung cancer groups were developed to identify the combinations of biomarkers that predict lung cancer in these cohorts.</p> <p>Results</p> <p>Statistically significant differences in the distributions of each of the biomarkers were identified among all five groups. Using Receiver Operating Characteristic (ROC) curves based on age, c-myc, Cyclin A, Cyclin B1, Cyclin D1, CDK2, and survivin, we obtained a sensitivity = 81% and specificity = 97% for the classification of cancer vs smokers(no nodules, solid nodules, or GGO) and correctly predicted 31/36 healthy controls as noncancer.</p> <p>Conclusion</p> <p>A pattern of autoantibody reactivity to TAAs may distinguish patients with lung cancer versus smokers with normal CTs, stable solid nodules, ground glass opacities, or normal healthy never smokers.</p

Glutamine depletion by crisantaspase hinders the growth of human hepatocellular carcinoma xenografts

Background: A subset of human hepatocellular carcinomas (HCC) exhibit mutations of β-catenin gene CTNNB1 and overexpress Glutamine synthetase (GS). The CTNNB1-mutated HCC cell line HepG2 is sensitive to glutamine starvation induced in vitro with the antileukemic drug Crisantaspase and the GS inhibitor methionine-L-sulfoximine (MSO). Methods: Immunodeficient mice with subcutaneous xenografts of the CTNNB1-mutated HCC cell lines HepG2 and HC-AFW1 were treated with Crisantaspase and/or MSO, and tumour growth was monitored. At the end of treatment, tumour weight and histology were assessed. Serum and tissue amino acids were determined by HPLC. Gene and protein expression were estimated with RT-PCR and western blot and GS activity with a colorimetric method. mTOR activity was evaluated from the phosphorylation of p70S6K1. Results: Crisantaspase and MSO depleted serum glutamine, lowered glutamine in liver and tumour tissue, and inhibited liver GS activity. HepG2 tumour growth was significantly reduced by either Crisantaspase or MSO, and completely suppressed by the combined treatment. The combined treatment was also effective against xenografts of the HC-AFW1 cell line, which is Crisantaspase resistant in vitro. Conclusions: The combination of Crisantaspase and MSO reduces glutamine supply to CTNNB1-mutated HCC xenografts and hinders their growth

No evidence of association between prothrombotic gene polymorphisms and the development of acute myocardial infarction at a young age

Background : we investigated the association between 9 polymorphisms of genes encoding hemostasis factors and myocardial infarction in a large sample of young patients chosen because they have less coronary atherosclerosis than older patients, and thus their disease is more likely to be related to a genetic predisposition to a prothrombotic state Methods and Results : this nationwide case-control study involved 1210 patients who had survived a first myocardial infarction at an age of 45 years who underwent coronary arteriography in 125 coronary care units and 1210 healthy subjects matched for age, sex, and geographical origin. None of the 9 polymorphisms of genes encoding proteins involved in coagulation (G-455A -fibrinogen: OR, 1.0; CI, 0.8 to 1.2; G1691A factor V: OR, 1.1; CI, 0.6 to 2.1; G20210A factor II: OR, 1.0; CI, 0.5 to 1.9; and G10976A factor VII: OR, 1.0; CI, 0.8 to 1.3), platelet function (C807T glycoprotein Ia: OR, 1.1; CI, 0.9 to 1.3; and C1565T glycoprotein IIIa: OR, 0.9; CI, 0.8 to 1.2), fibrinolysis (G185T factor XIII: OR, 1.2; CI, 0.9 to 1.6; and 4G/5G plasminogen activator inhibitor type 1: OR, 0.9; CI, 0.7 to 1.2), or homocysteine metabolism (C677T methylenetetrahydrofolate reductase: OR, 0.9; CI, 0.8 to 1.1) were associated with an increased or decreased risk of myocardial infarction Conclusions : this study provides no evidence supporting an association between 9 polymorphisms of genes encoding proteins involved in hemostasis and the occurrence of premature myocardial infarction or protection against it

Veicolo terrestre provvisto di almeno un’appendice aerodinamica

La presente invenzione ha per oggetto un veicolo terrestre (A) comprendente almeno un portamozzo (15) per il mozzo di una ruota (13), una massa sospesa (16) rispetto al portamozzo (15,15’) per mezzo di un elemento elastico (17,17’) ed una appendice aerodinamica (2,2’) atta a generare una opportuna forza aerodinamica in condizione di veicolo in movimento. L’appendice aerodinamica (2,2’) è collegata al portamozzo (15,15’) e alla massa sospesa (16) attraverso almeno un sistema articolato (1,1'). Quest’ultimo comprende almeno una leva (6) infulcrata sulla massa sospesa (16) ed accoppiata al portamozzo (15,15’), ed almeno un organo di trasmissione (5,6,7,8,9,10) collegato a detta appendice aerodinamica (2,2’) e a detta leva (6) in modo tale da applicare una forza verticale risultante in un punto di detta leva (6)

Il dolore nel parto: l\u2019assistenza ostetrica al parto con e senza analgesia

Il dolore del parto \ue8 un aspetto nodale, non solo nel vissuto della donna che vive il travaglio e il parto, ma anche nel bagaglio di competenze degli operatori che lavorano in questo campo, le ostetriche in particolare. Il momento storico che stiamo vivendo rende pi\uf9 urgente la necessit\ue0 di una riflessione a questo proposito, non tanto perch\ue8 l\u2019ingresso in sala parto dell\u2019analgesia epidurale costituisce ancora una novit\ue0 per molte realt\ue0 ospedaliere italiane, quanto per la diffusione di un pensiero secondo cui il parto necessita di metodi analgesici, che siano questi naturali o meno. La soddisfazione di ogni donna rispetto all\u2019esperienza della nascita non dipende dall\u2019assenza o dalla riduzione del dolore: si tratta di un dato riconosciuto dalla letteratura e confermato dai racconti delle puerpere. Lo sforzo degli operatori che si occupano del percorso nascita non dovrebbe essere rivolto tanto a valorizzare le tecniche analgesiche, quanto a favorire che le sensazioni dolorifiche siano tollerate nel modo migliore, unico per ogni partoriente. Su questo intento deve essere convogliata tutta la competenza creativa dell\u2019ostetrica ma anche la sua capacit\ue0 diagnostica. L\u2019ostetrica, infatti, deve essere in grado di leggere i segnali corporei delle donne in modo da consigliarle, sostenerle, ed aiutarle a trovare strategie spontanee per affrontare la difficolt\ue0 del travaglio distinguendo tra il dolore fisiologico dei parti senza complicazioni e il dolore \u201canormale\u201d, associato a travagli distocici, malposizioni fetali o ad interventi iatrogeni. I fattori pi\uf9 rilevanti nel determinare la soddisfazione delle donne hanno a che fare con: - Le aspettative delle donne; - Il sostegno offerto e la qualit\ue0 della relazione che si instaura con l\u2019ostetrica; - La partecipazione della donna; - La continuit\ue0 dell\u2019assistenza