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

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

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

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

Structural Basis of Affinity Maturation of Antibodies in the 2- Phenyl-5-Oxazolone System

Background: Affinity maturation of antibodies is the process whereby more efficient antibodies are produced through somatic hypermutation and antigen-guided selection. No extensive study is available at the moment concerning the relationship between somatic mutations and their structural counterpart. The antibody response to 2-phenyl-5-oxazolone has been thoroughly investigated from the genetic point of view. It consists of three antibody classes, with each member of each class derived from a unique pair of VH and VL germline genes by somatic hypermutation. In this project, we are investigating the structure of the VH and VL domains of 10 representative antibodies. Methods: The VH and VL domains of each antibody are being expressed as recombinant scFvs, crystallised, and their structure determined by X-ray crystallography. Results: The structures and models available allow an initial definition of the strategies adopted. In class I, maturation is bound to improvement of surface complementarity, especially at the top of the binding pocket, and in surface charge changes. In class II the maturation strategy seems to be based on the increase of the interacting surface, and on the introduction of a specific bond with the oxazolone ring. In class III, where the low and high affinity antibodies differ by 8 mutations, the increase in affinity is mainly determined by the improvement in the surface complementarity by removal of a bulky phenylalanine, which allows a better tightening of the two sides of the binding site. Conclusions: These results are relevant to determine the principles underlying affinity maturation of antibodies

Pharmacokinetics of a Sustained Release Formulation of Pyridoxal Phosphate of Buflomedil After Single or Repeated Oral Doses in Healthy Volunteers.

The pharmacokinetics of a sustained release (SR) formulation of pyridoxal phosphate of buflomedil (Pirxane retard) has been studied after oral administration to healthy volunteers using among else a gaschromatographic dosage method. After oral administration of 400 mg of the SR formulation, pyridoxal phosphate of buflomedil has a much slower kinetics compared to the normal formulation (tmax:approx. 1.5 h) reaching the maximum plasma concentration, which was about 467 ng/ml, in about 3 h. After 24 h the concentrations were still about 1/10 (48 ng/ml) the maximum value. 24-h urinary excretion was about 21% of the administered dose. Repeated administration of the SR formulation for 7 days in single daily doses of 400 mg gave steady state plasma levels (ca. 250 ng/ml) 12 h after the administration without statistically significant variations. The plasma concentrations of the drug measured daily after reaching the steady state were similar one to the other. The tolerability was very good and no local or systemic side effects of any kind were reported