MITO END-3: Efficacy of Avelumab immunotherapy according to molecular profiling in first-line endometrial cancer therapy.

BACKGROUND: Immunotherapy combined with chemotherapy significantly improves progression-free survival compared to first-line chemotherapy alone in advanced endometrial cancer, with a much larger effect size in microsatellite-instability high (MSI-H) cases. New biomarkers might help to select patients that may have benefit among those with a microsatellite-stable (MSS) tumor. METHODS: In a pre-planned translational analysis of the MITO END-3 trial, we assessed the significance of genomic abnormalities in patients randomized to standard carboplatin/paclitaxel without or with avelumab. RESULTS: Out of 125 randomized patients, 109 had samples eligible for next-generation sequencing (NGS) analysis, and 102 had MSI tested. According to The Cancer Genome Atlas (TCGA), there were 29 cases MSI-H, 26 MSS TP53 wild-type (wt), 47 MSS TP53 mutated (mut), and one case with POLE mutation. Four mutated genes were present in more than 30% of cases: TP53, PIK3CA, ARID1A, and PTEN. Eleven patients (10%) had a BRCA1/2 mutation (five in MSI-H and six in MSS). High TMB (≥10 Muts/Mb) was observed in all MSI-H patients, in four out of 47 MSS/TP53 mut, and no case in the MSS/TP53 wt category. The effect of avelumab on progression-free survival significantly varied according to TCGA categories, being favorable in MSI-H and worst in MSS/TP53 mut (P interaction=0.003); a similar non-significant trend was seen in survival analysis. ARID1A and PTEN also showed a statistically significant interaction with treatment effect, which was better in the presence of the mutation (ARID1A P interaction=0.01; PTEN P interaction=0.002). CONCLUSION: The MITO END-3 trial results suggest that TP53 mutation is associated with a poor effect of avelumab, while mutations of PTEN and ARID1A are related to a positive effect of the drug in patients with advanced endometrial cancer.

Alternative academic approaches for testing homologous recombination deficiency in ovarian cancer in the MITO16A/MaNGO-OV2 trial.

BACKGROUND: The detection of homologous recombination deficiency (HRD) can identify patients who are more responsive to platinum and poly ADP ribose polymerase inhibitors (PARPi). MyChoice CDx (Myriad) is the most used HRD test in ovarian cancer (OC). However, some limitations of commercial tests exist, because of the high rate of inconclusive results, costs, and the impossibility of evaluating functional resistance mechanisms. PATIENTS AND METHODS: Two academic genomic tests and a functional assay, the RAD51 foci, were evaluated to detect HRD. One hundred patients with high-grade OC enrolled in the MITO16A/MaNGO-OV2 trial and treated with first-line therapy with carboplatin, paclitaxel, and bevacizumab were analyzed. RESULTS: The failure rate of the two genomic assays was 2%. The sensitivity in detecting HRD when compared with Myriad was 98.1% and 90.6%, respectively. The agreement rate with Myriad was 0.92 and 0.87, with a Cohen's κ coefficient corresponding to 0.84 and 0.74, respectively. For the RAD51 foci assay, the failure rate was 30%. When the test was successful, discordant results for deficient and proficient tumors were observed, and additional HRD patients were identified compared to Myriad; sensitivity was 82.9%, agreement rate was 0.65, and Cohen's κ coefficient was 0.18. The HRD detected by genomic assays and residual tumor at primary surgery and stage was correlated with progression-free survival at multivariate analysis. CONCLUSIONS: Results suggest the feasibility of academic tests for assessing HRD status that show robust concordance with Myriad and correlation with clinical outcome. The contribution of the functional information related to the RAD51 foci test to the genomic data needs further investigation.

Intestinal-type mucinous ovarian carcinoma arising from a seromucinous precursor lesion.

OBJECTIVE: Mucinous ovarian carcinoma is a tumor with gastrointestinal differentiation, which is not associated with endometrial-type (endometriotic or seromucinous) precursors. Here, we describe a peculiar case of mucinous ovarian tumor with intestinal differentiation arising in a seromucinous lesion, which may represent a distinct entity. CASE PRESENTATION: A 58-year woman underwent surgery due to a 14.5-cm ovarian mass with lymph nodal, peritoneal, omental and colorectal involvement. Histological examination with ancillary immunohistochemical analysis has been performed. Histologically, the mass was a carcinoma with intestinal differentiation and expansile growth pattern, arising in a seromucinous cystadenoma with intestinal metaplasia. Both the carcinoma and the metaplasia showed loss of Müllerian markers (estrogen and progesterone receptors, PAX8) and positivity for intestinal-type markers (cytokeratin 20, CDX2). CONCLUSIONS: Our case may represent the ovarian counterpart of endometrial gastrointestinal-type carcinoma, which is an aggressive entity developing from gastrointestinal metaplasia of the endometrial epithelium. Acknowledging the existence of such entity might be relevant in terms of diagnosis and patient management.

The type 2B p.R1306W natural mutation of von Willebrand factor dramatically enhances the multimer sensitivity to shear stress.

BACKGROUND: Shear stress triggers conformational stretching of von Willebrand factor (VWF), which is responsible for its self-association and binding to the platelet receptor glycoprotein (GP)Ibα. This phenomenon supports primary hemostasis under flow. Type 2B VWF natural mutants are considered to have increased affinity for platelet GPIbα. OBJECTIVES: To assess the mechanism responsible for the enhanced interaction of the p.R1306W VWF mutant with the platelet receptor. METHODS: The interaction of GPIbα with wild-type (WT) and p.R1306W VWF multimers and A1-A2-A3 constructs was investigated with surface plasmon resonance spectroscopy. Analysis of the static VWF conformation in solution was performed with dynamic light scattering spectroscopy. The shear stress-induced self-association of VWF multimers was investigated with atomic force microscopy (AFM) over a 0-60 dyn cm(-2) range. RESULTS: WT VWF did not interact with GPIbα under static conditions, whereas the mutant at ~ 2 µg mL(-1) already bound to the receptor. By contrast, the WT and p.R1306W-A1-A2-A3 constructs showed comparable affinities for GPIbα (Kd  ~ 20 nm). The hydrodynamic diameter of resting R1306W VWF multimers was significantly greater than that of the wild type (210 ± 60 nm vs. 87 ± 22 nm). At shear forces of < 14 dyn cm(-2) , the p.R1306W multimers rapidly changed conformation, entering a regime of self-aggregation, which, in contrast, was induced for WT VWF by shear forces of > 30 dyn cm(-2) . Mechanical stretching AFM experiments showed that p.R1306W multimers needed less energy per length unit (~ 10 pN) to be stretched than the WT protein. CONCLUSIONS: The increased affinity of p.R1306W VWF for GPIbα arises mostly from higher sensitivity to shear stress, which facilitates exposure of GPIbα binding sites.

ß2 -Glycoprotein I binds to thrombin and selectively inhibits the enzyme procoagulant functions.

BACKGROUND: This work was aimed at characterizing the interaction of ß(2)-glycoprotein I (ß(2)GPI), an abundant plasma protein of unknown function, with human thrombin, the final effector protease in the coagulation cascade. METHODS: The ß(2)GPI-thrombin interaction was studied by surface plasmon resonance (SPR), fluorescence, and molecular modeling. The effect of ß(2)GPI on the procoagulant (fibrin generation and platelet aggregation) and anticoagulant (protein C activation) functions of thrombin were investigated with turbidimetric, immunocytofluorimetric and enzymatic assays. RESULTS: SPR and fluorescence data indicated that ß(2)GPI tightly bound thrombin (K(d) = 34 nM) by interacting with both protease exosites, while leaving the active site accessible. This picture is fully consistent with the theoretical model of the ß(2)GPI-thrombin complex. In particular, blockage of thrombin exosites with binders specific for exosite-1 (hirugen and HD1 aptamer) or exosite-2 (fibrinogen γ'-peptide and HD22 aptamer) impaired the ß2 GPI-thrombin interaction. Identical results were obtained with thrombin mutants having one of the two exosites selectively compromised by mutation (Arg73Ala and Arg101Ala). Fluorescence measurements indicated that ß(2)GPI did not affect the affinity of the enzyme for active site inhibitors, such as p-aminobenzamidine and the hirudin(1-47) domain, in agreement with the structural model. ß(2)GPI dose-dependently prolonged the thrombin clotting time and ecarin clotting time in ß(2)GPI-deficient plasma. ß(2)GPI inhibited thrombin-induced platelet aggregation (IC50 = 0.36 µM) by impairing thrombin cleavage of protease-activated receptor 1 (PAR1) (IC50 = 0.32 µM), both on gel-filtered platelets and in whole blood. Strikingly, ß(2) GPI did not affect thrombin-mediated generation of the anticoagulant protein C. CONCLUSIONS: ß(2) GPI functions as a physiologic anticoagulant by inhibiting the key procoagulant activities of thrombin without affecting its unique anticoagulant function.

Oxidized von Willebrand factor is efficiently cleaved by serine proteases from primary granules of leukocytes: divergence from ADAMTS-13.

BACKGROUND: The leukocyte serine proteases (LSPs) elastase, proteinase 3 and cathepsin G cleave von Willebrand factor (VWF) near or at the same cleavage site (Tyr1605-Met1606) as ADAMTS-13, the metalloprotease that specifically controls the proteolytic processing of VWF. Recent studies have shown that oxidation of VWF at Met1606 with formation of methionine sulfoxide (MetSO) severely impairs its proteolysis by ADAMTS-13. METHODS: This study was aimed at assessing whether or not oxidation of VWF by reactive oxygen species (ROS) can also affect its cleavage by elastase, proteinase 3, and cathepsin G. In this study, the catalytic specificity of hydrolysis by LSPs of the VWF peptide substrate VWF74 and full-length VWF, both unaltered and in the oxidized form, was measured by RP-HPLC, electrophoretic and mass spectrometry methods. RESULTS: LSPs cleaved both VWF multimers and VWF74 near or at the same peptide bond as is cleaved by ADAMTS-13, with k(cat)/K(m) values similar to those of the metalloprotease. However, unlike ADAMTS-13, cathepsin G cleaved VWF74 containing a MetSO residue at position 1606 with a k(cat)/K(m) value higher than that for VWF74, whereas the catalytic efficiencies of both elastase and proteinase 3 were unaffected by the replacement of Met1606 with MetSO. Likewise, oxidation of VWF multimers by hypochlorous acid and ROS, produced by activated leukocytes, improved their hydrolysis by LSPs. CONCLUSIONS: Oxidation by leukocyte ROS has a net positive effect on the cleavage of VWF multimers by LSPs, under conditions where high concentrations of oxidant species would severely reduce the proteolytic efficiency of ADAMTS-13.

Isolation of Gram-positive n-alkane degraders from a hydrocarbon-contaminated Mediterranean shoreline.

AIMS: To investigate the petroleum hydrocarbon (HC)-degrading potential of indigenous micro-organisms in a sandy Mediterranean coast, accidentally contaminated with petroleum-derived HCs. METHODS AND RESULTS: Using culturable methods, a population of Gram-positive n-alkane degraders was detected in the contaminated soil. Five isolates, identified as one Nocardia, two Rhodococcus and two Gordonia strains, were able to degrade medium- and long-chain n-alkanes up to C(36) as assessed by growth assays and gas chromatography-mass spectrometry analysis. Diverging alkane hydroxylase-encoding genes (alkB) were detected by PCR, using degenerated primers, in all the strains; multiple sequences were obtained from the Nocardia strain, while only one alkB gene was detected in the Rhodococcus and Gordonia strains. The majority of the alkB sequences were related to Rhodococcus alkB2, but none was identical to it. CONCLUSIONS: Actinomycetes might have a key role in bioremediation of n-alkane-contaminated sites under dry, resource-limited conditions, such as those found in the Mediterranean shorelines. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first study on the bioremediation potential in Mediterranean contaminated beaches.

Long-acting octreotide as rescue therapy in chronic bleeding from gastrointestinal angiodysplasia.

BACKGROUND: Octreotide has shown to be effective against rebleeding from gastrointestinal angiodysplasias, but a long-term daily parenteral administration is recommended. Long-acting octreotide (LAR-OCT) could overcome such a limitation, but it has not been studied extensively. AIM: To investigate the usefulness of long-acting octreotide in the control of chronic bleeding from gastrointestinal angiodysplasias. METHODS: Thirteen patients with chronic gastrointestinal bleeding because of angiodysplasias were enrolled. Diagnosis was made by endoscopy and wireless video capsule. Long-acting octreotide was administered intramuscularly at a dosage of 10 mg/monthly for 1 year. Patients were followed up for a minimum period of 1 year, and haemoglobin levels, blood transfusions, iron supplementation and hospitalizations were recorded 1 year before and after starting long-acting octreotide therapy. RESULTS: Follow-up ranged from 12 to 60 months. Nine of 13 patients (69%) did not need blood transfusions and iron supplementation any longer; a partial improvement was observed in one patient; no effect was found in the others. No side effect was recorded in any patient. CONCLUSIONS: Long-acting octreotide for 1 year may be beneficial as a rescue therapy for controlling chronic bleeding from gastrointestinal angiodysplasias in patients not eligible for surgery. Its monthly administration represents an advantage, which makes such a formulation the choice when a long-term treatment is mandatory.

Bradyrhizobium sp. nodulating the Mediterranean shrub Spanish broom (Spartium junceum L.).

AIMS: The molecular diversity of 25 strains of rhizobia, isolated in Sicily from root nodules of the Mediterranean shrubby legume Spanish broom (Spartium junceum L.), is presented in relation to the known rhizobial reference strains. METHODS AND RESULTS: Our approach to the study of the S. junceum rhizobial diversity combined the information given by the 16S and the intergenic spacer (IGS) 16S-23S rDNA polymorphic region by obtaining them in a single polymerase chain reaction (PCR) step. The PCR fragment size of the S. junceum isolates was 2400-2500 bp and that of the reference strains varied from 2400 in Bradyrhizobium strains to 2800 in Sinorhizobium strains. Inter- and intrageneric length variability was found among the reference strains. Restriction fragment length polymorphisms (RFLP) analysis allowed us to identify eight genotypes among the S. junceum rhizobia that were clustered into two groups, both related to the Bradyrhizobium lineage. Sequencing of representative strains of the two clusters confirmed these data. The 16S-IGS PCR-RFLP approach, when applied to rhizobial reference strains, allowed very close species (i.e. Rhizobium leguminosarum/R. tropici) to be separated with any of the three enzymes used; however, cluster analysis revealed inconsistencies with the 16S-based phylogenesis of rhizobia. CONCLUSIONS: Rhizobia nodulating S. junceum in the Mediterranean region belong to the Bradyrhizobium lineage. Our results confirm the resolution power of the 16S-23S rDNA in distinguishing among rhizobia genera and species, as well as the usefulness of the PCR-RFLP method applied to the entire 16S-IGS region for a rapid tracking of the known relatives of new isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: The present paper is, to our knowledge, the first report on rhizobia nodulating a Mediterranean wild woody legume.

The prognostic value, in active ulcerative colitis, of an increased intensity of colonic perivisceral fat signal on magnetic resonance imaging with ferumoxil.

OBJECTIVE: In active ulcerative colitis (UC), magnetic resonance imaging (MRI) with ferumoxil, a superparamagnetic oral contrast agent, accurately evaluates, in our experience, the increased wall thickness and frequently shows a stronger perivisceral fat signal intensity (PFSI). The aim of our study was to evaluate the clinical significance of these MRI findings in active UC. METHODS: Twenty-four consecutive patients affected by moderate pancolitis were enrolled. At entry, each patient underwent MRI with ferumoxil to evaluate wall thickness and PFSI. Two groups of patients were individuated: group A (increased PFSI) and group B (normal PFSI). After obtaining remission, the number of relapses and, at each flare-up, the clinical activity index (CAI) were evaluated in all patients in a 2-yr follow-up period. The mean CAI was calculated at the end of the follow-up in each patient. Where there was colectomy, a complete histological examination of the colon was performed. RESULTS: PFSI was increased in 16 patients (group A) and was normal in the remainder (group B). There was a significant difference of wall thickness, number of relapses/yr, and mean CAI between the two groups of patients. No difference was observed with regard the duration of disease. Six patients of group A and no patient of group B underwent colectomy. The histological evaluation showed an increased thickness of the entire colonic wall with significant changes of the perivisceral fat structures. CONCLUSIONS: An increased

Allergy and mucosal eosinophil infiltrate in ulcerative colitis.

BACKGROUND: Data on allergy in ulcerative colitis (UC) have led to conflicting conclusions without proving any causal association. In this report we have investigated the presence of allergy and its possible relation with chronic colonic inflammation in patients with UC. METHODS: Fifty UC patients underwent clinical, endoscopic, and histologic evaluations. The allergologic study included family/personal history; prick/patch exposition to airborne, food, and contact allergens; total serum IgE; and quantification of eosinophils in peripheral blood and intestinal mucosa. Diagnosis of rhinitis, conjunctivitis, and asthma was confirmed by specific provocation tests. Fifty healthy subjects were studied as control group. RESULTS: A higher prevalence of allergic symptoms was found in patients (56%) and their first-degree relatives (52%) than in controls (18% and 26%) (P < 0.0001; P = 0.008). In patients skin tests showed increased rates of immediate (54%) and delayed-type (20%) hypersensitivity compared with controls (30% and 6%) (P= 0.01; P= 0.03). Diagnosis of allergic IgE-mediated disease was made in 19 cases and 6 controls (P= 0.01), and allergic contact dermatitis in 10 and 3, respectively (P= 0.03). IgE levels were higher in UC patients than in controls (P=0.02). No dose-response relationship was found between degree of colonic tissue eosinophilia and clinical. endoscopic, and histologic disease severity. The degree of colonic tissue eosinophilia was higher in the presence of skin reactivity to food allergens. CONCLUSIONS: UC patients frequently show several markers of allergy. In particular, our data suggest an association between ulcerative colitis, tissue eosinophilia, and type-I allergy.

Magnetic resonance imaging with ferumoxil, a negative superparamagnetic oral contrast agent, in the evaluation of ulcerative colitis.

OBJECTIVE: The introduction of new oral contrast agents that enhance image quality has increased the importance of magnetic resonance imaging (MRI) in the management of ulcerative colitis. The aim of our study was to investigate the usefulness of a new negative superparamagnetic oral contrast (ferumoxil) alone or in association with gadolinium i.v. in the assessment of the disease. METHODS: Twenty-eight patients with clinically active ulcerative colitis and 10 control subjects entered the study. In each patient a clinical, endoscopic, histological, and MRI evaluation was performed. In particular, in 14 patients affected by ulcerative colitis (group A) and in five controls, magnetic resonance images were acquired 1 h after the oral administration of 900 ml of ferumoxil, while the remaining 14 patients (group B) and five controls were submitted to double-contrast MRI (ferumoxil and gadolinium). In both groups, wall thickness, length of affected bowel segments, and, in group B, also percent contrast enhancement were calculated. RESULTS: The comparison of endoscopic and MRI extent of disease was statistically significant. Wall thickness and, in group B, also percent contrast enhancement were significantly correlated with clinical and endoscopic activities. In each group wall thickness was significantly different in the activity phases of the disease. CONCLUSIONS: MRI with negative superparamagnetic oral contrast is comparable to endoscopy in the assessment of ulcerative colitis. The double-contrast imaging does not provide more information than single oral contrast, so we concluded that the latter is preferable in the follow-up of the disease and in patients unable or with a poor compliance to undergo endoscopy.

Prognostic value of progressive decrease in serum cholesterol in predicting survival in Child-Pugh C viral cirrhosis.

BACKGROUND: The identification of cirrhotic patients with low life expectancy is an open clinical problem. Hypocholesterolemia is frequently found in severe chronic hepatic insufficiency because the liver is the most active site of cholesterol metabolism, but poor information is available on its precise prognostic value. We evaluated the prognostic role of hypocholesterolemia in patients with advanced liver cirrhosis. METHODS: Serial serum cholesterol concentrations of 34 patients with virus-induced cirrhosis, from the first appearance of Child-Pugh class C to death, were considered. To compare survival functions, we established three base-line cholesterol cut-off points (150, 125, and 100 mg/dl) and stratified patients into groups A and B, with base-line cholesterol levels lower and higher than each cut-off value, respectively. RESULTS: Cholesterolemia decreased progressively in all patients. At the 100 mg/dl cut-off point all group-A patients died within 17 months, whereas 75% of group-B patients were alive at 24 months (P < 0.0001). Moreover, cholesterolemia was significantly correlated with cholinesterase, indirect bilirubin, and total bilirubin at entry time and immediately before death. No correlation was observed between cholesterol and these variables when stratified for the Child-Pugh score. CONCLUSIONS: Base-line serum cholesterol levels lower than 100 mg/dl identify a subgroup of Child-C cirrhotic patients with high mortality risk within a 2-year follow-up. The prognostic importance of cholesterolemia may also be deduced by the significant correlation with other well-established indicators of survival.

Beclomethasone dipropionate (3 mg) enemas combined with oral 5-ASA (2.4 g) in the treatment of ulcerative colitis not responsive to oral 5-ASA alone.

BACKGROUND/AIMS: Beclomethasone dipropionate is one of the topical corticosteroids which appear to have minimal systemic effects. We evaluated whether combined therapy with Beclomethasone dipropionate enemas and oral 5-aminosalicylic acid could be effective in patients suffering from ulcerative colitis not responsive to oral 5-aminosalicylic acid as monotherapy. PATIENTS: In twenty patients, non responders to 5-aminosalicylic acid treatment (2.4-3.6 g/day) given for at least 6 weeks, Beclomethasone dipropionate enemas (3 mg/60 ml/day) were added for 4 weeks. METHODS: Efficacy of the combination was evaluated before and at the end of the treatment using a clinical, endoscopic and histological score. RESULTS: After a four-week treatment period, a significant clinical improvement in stool frequency (p < 0.01), stool consistency (p < 0.001), blood (p < 0.001) and mucus in stools (p < 0.05), was observed. Endoscopy and biopsy confirmed an improvement in the activity score at the end of the treatment (p < 0.001). Six patients (30%) achieved remission, ten patients showed an improvement (50%) and four (20%) showed no benefits. No adverse event was observed. CONCLUSIONS: Beclomethasone dipropionate enemas combined with oral 5-aminosalicylic acid may be a safe and useful therapeutic approach in the treatment of ulcerative colitis not responsive to oral 5-aminosalicylic acid alone.

Reemergence of Shigella dysenteriae type 2 in Sicily: an epidemiological evaluation.

Three strains of Shigella dysenteriae type 2 were isolated from a small household outbreak which occurred in Palermo, Sicily, during summer 1990. Two isolates were recovered from hospitalized patients and one from an asymptomatic carrier. The infection could not be associated with travel to foreign countries or contact with travellers returned from abroad. Since 1953 S. dysenteriae has been never isolated in Southern Italy. The isolates from dysentery cases were susceptible to antibiotics and carried a plasmid of 120 MDa associated with a small cryptic plasmid; in contrast, the strain isolated from the healthy carrier contained an additional plasmid of approximately 40 MDa, which codified for resistance to ampicillin, streptomycin, sulfamethoxazole and trimethoprim. All strains showed some atypical biochemical properties, but their rRNA-DNA patterns of hybridization were closely similar to that of the reference strains of type 2 and easily distinguishable from those of the other types of non-Shiga bacillus reference strains. Epidemiological isolation features of these strains suggest a possible circulation of this Shigella species in Sicily. Genetic characterization of these strains may be useful for surveillance of infections by this organism.

New perspectives for the therapeutic management of recurrent immunological abortion.

Up to now the only effective therapy for recurrent abortion syndrome due to the absence of the so-called blocking-factor has been active immunotherapy with partner or third-party donor mononucleates. The Authors report their in vivo and in vitro experience with high-dose intravenous gammaglobulin (i IV Ig) in order to treat women with recurrent abortion syndrome. In the Authors opinion there are sufficient experimental reasons for continuing this research with IV Ig obtained from multiparous women plasma pools.

Immunological problems in the recurrent abortion syndrome.

The recurrent abortion syndrome has been considered a serious obstetrical problem, since it was not possible to diagnose the causes in over 40% of the cases. Great progress has now been made with the knowledge of the immunological mechanisms involved in the maintenance of pregnancy. The disorders of such immunological mechanisms, both due to auto- or alloimmune problems, are demonstrated in about 40% of the cases of recurrent abortions. The Authors have studied a group of 56 patients with recurrent abortion syndrome in a complex diagnostic work-up involving: 1) The research of an autoimmune cause; 2) The research of the presence of the Blocking-factors by means of one-way mixed lymphocyte cultures; 3) The research of an altered antipaternal lymphocytotoxic activity or of an excessive HLA-sharing. According to the results of the investigations, several forms of therapeutic management have been used: 1) Steroids-aspirin-calcic heparin for Autoimmune cases. 2) Active immunotherapy for the cases due to the lack of blocking factors. 3) Calcic heparin for the altered antipaternal lymphocytotoxic activity. At the moment it is difficult to evaluate the results of pregnancy rate with this type of therapeutic management because of the too short follow-up.