'Atherothrombosis-associated microRNAs in Antiphospholipid syndrome and Systemic Lupus Erythematosus patients'.

MicroRNAs markedly affect the immune system, and have a relevant role in CVD and autoimmune diseases. Yet, no study has analyzed their involvement in atherothrombosis related to APS and SLE patients. This study intended to: 1) identify and characterize microRNAs linked to CVD in APS and SLE; 2) assess the effects of specific autoantibodies. Six microRNAs, involved in atherothrombosis development, were quantified in purified leukocytes from 23 APS and 64 SLE patients, and 56 healthy donors. Levels of microRNAs in neutrophils were lower in APS and SLE than in healthy donors. Gene and protein expression of miRNA biogenesis-related molecules were also reduced. Accordingly, more than 75% of identified miRNAs by miRNA profiling were underexpressed. In monocytes, miR124a and -125a were low, while miR-146a and miR-155 appeared elevated. Altered microRNAs' expression was linked to autoimmunity, thrombosis, early atherosclerosis, and oxidative stress in both pathologies. In vitro treatment of neutrophils, monocytes, and ECs with aPL-IgG or anti-dsDNA-IgG antibodies deregulated microRNAs expression, and decreased miRNA biogenesis-related proteins. Monocyte transfections with pre-miR-124a and/or -125a caused reduction in atherothrombosis-related target molecules. In conclusion, microRNA biogenesis, significantly altered in neutrophils of APS and SLE patients, is associated to their atherothrombotic status, further modulated by specific autoantibodies.

Identification of miRNAs as potential modulators of tissue factor expression in patients with systemic lupus erythematosus and antiphospholipid syndrome.

BACKGROUND: Tissue factor (TF) is the main initiator of the coagulation cascade and elements that may upregulate its expression might provoke thrombotic events. Systemic lupus erythematosus (SLE) and antiphospholipid syndrome (APS) are autoimmune diseases characterized by a high TF expression in monocytes. OBJECTIVES: To examine the role of microRNAs (miRNAs) in TF expression and to evaluate their levels in SLE and APS patients. METHODS: An in silico search was performed to find potential putative binding sites of miRNAs in TF mRNA. In vitro validation was performed transfecting cells expressing TF (THP-1 and MDA-MB-231) with oligonucleotide miRNA precursors and inhibitors. Additionally, reporter assays were performed to test for the binding of miR-20a to TF mRNA. Levels of miRNAs and TF were measured by quantitative (qRT-PCR) in patients with APS and SLE. RESULTS: Overexpression of miRNA precursors, but not inhibitors, of two of the members of cluster miR-17∼92, for example miR-19b and miR-20a, in cells expressing TF decreased TF mRNA, protein levels, and procoagulant activity between 30% and 60%. Reporter assays showed that miR-20a binds to TF mRNA. Finally, we measured levels of miR-19b and miR-20a in monocytes from patients with APS and SLE and observed significantly lower miRNAs levels in comparison with healthy subjects inversely correlated with the levels of TF. CONCLUSIONS: Down-regulation of miR-19b and miR-20a observed in patients with SLE and APS could contribute to increased TF expression and thus provoke the hypercoagulable state characteristic of these patients.

Rare homozygous status of P43 ß1-tubulin polymorphism causes alterations in platelet ultrastructure.

ß1-tubulin is the main constituent of the platelet marginal band and studies with deficient mice showed that it maintains discoid shape and it is required for normal platelet formation. TUBB1 Q43P polymorphism is associated with decreased ß1-tubulin expression, diminished platelet reactivity, and partial loss of discoid shape in heterozygous carriers. However, to date no studies have been carried out on homozygous PP individuals. Our study included 19 subjects genotyped for TUBB1 Q43P polymorphism (4 QQ, 4 QP, and 2 PP). The two PP individuals were recruited after genotyping of 2073 individuals. Biochemical, microscopy, and molecular studies were performed. Real-time PCR showed a ~40% decrease in TUBB1 mRNA in the two PP individuals compared to four QQ subjects. Western blot analysis confirmed this reduction. Electron microscopy revealed a majority of normal discoid platelets in PP individuals, although platelets with loose, re-orientated or invaginated protofilaments, and an over-developed open canalicular system were observed. Such abnormalities were not observed in QQ subjects. Morphometric analyses showed no differences between PP and QQ individuals. Immunofluorescence confirmed the presence of a normal marginal band in a majority of platelets from PP subjects. Interestingly, both PP subjects had a 40% lower platelet count than QP and QQ. TUBB1 Q43P polymorphism in homozygosity mildly affects platelet ultrastructure and our data further suggest that high levels of ß1-tubulin might not be critical to sustain platelet discoid shape.

Effects of acrolein, a natural occurring aldehyde, on the anticoagulant serpin antithrombin.

We studied the effect of acrolein, an alpha,beta-unsaturated aldehyde that causes adduct-modification of lysine, cysteine, and histidine residues, on antithrombin, a key anticoagulant serpin. Intrinsic fluorescence, functionality (anti-FXa and anti-IIa activity), heparin affinity and conformational features of plasma and purified antithrombin were evaluated. In vivo experiments were carried out in mice. Intrinsic fluorescence showed a two-step conformational change. Acrolein, even at low dose, impaired the anticoagulant function of purified antithrombin by affecting its heparin affinity. However, higher concentrations of acrolein and long incubations are required to cause mild functional effects on plasma antithrombin and mice.

Primary tubal choriocarcinoma.

Choriocarcinoma is one of the most serious forms of gestational trophoblastic tumor. It is a malignant tumor from the epithelium of the chorionic villi. The most frequent location site is the uterus. Associated with ectopic pregnancy, it is extremely rare and in general, very aggressive. In 75% of the cases, it items from distant metastasis; therefore, a histological examination of the tubes must be performed in all ectopic pregnancies. Our patient was a 33-year-old woman who was admitted to emergency room (ER) with an intense pain in the right, iliac cavity, and limited genital bleeding. During the exploration, there was abdominal pain, with doubtful signs of peritoneal irritation. The vaginal ultrasound offered an image that was compatible with an extra uterine pregnancy in the left appendages. At emergency, right salpingectomy was performed via laparotomy. The patient was treated with polychemotherapy and contraceptives for a year, with no recurrence of the disease. Control follow-up was performed using beta-human chorionic gonadotropin (HCG) testing on a weekly basis during the first month and then bi-monthly during the first year of follow-up.

[Glandular cancer of the cervix in the initial stages. Treatment and results at the Curie Institute]. / Cancer glandulaire du col utérin aux stades initiaux. Traitements et résultats à l'Institut Curie.

102 patients were treated for adenocarcinoma of the cervix, stage 1 and 2, by radiotherapy and surgery at the Curie Institute between the years 1955 and 1974. Glandular carcinoma represents about 10% of cancers of the uterine cervix. The survival rate is: at five years 81%, at ten years 78%, at fifteen years 74%. This is similar to the survival rate for squamous carcinoma of the same stages. 48% of adenocarcinomata are found to be sterilised on the surgical specimen as compared with 78% in squamous cell carcinomata. This justified the even more frequent use of radiotherapy with surgery in this type of tumour.

[Sequential test with clomiphene and LH-RH in cases of anovulatory sterility (author's transl)]. / Le test séquentiel clomifène-LH-RH en cas de stérilité due à un cycle anovulatoire.

In sterile menstruating women with anovulatory cycle, the response of plasmatic gonadotropins and urinary total Estrogens and Pregnandiol to the administration of Clomifene and LH-RH in a sequential test is studied. Five different types of response, similar to those detected in previous studies using the LH-RH test, are identified; in them, uniformity in the response of gonadotropins exists to both LH-RH and Clomifene. The results obtained seem to confirm the existence of diverse etiopathogenic mechanisms in the alterations of the Hypothalamus-Pituitary-Ovarian axis resulting in anovulatory cycle.