Spectrum of F8 gene mutations in haemophilia A patients from a region of Italy: identification of 23 new mutations.

SUMMARY: Haemophilia A (HA) is an X-linked recessive bleeding disorder caused by a lack or decrease of coagulation factor VIII activity. The molecular diagnosis of HA is challenging and a variety of different mutations have been identified throughout the F8 gene. Our aim was to detect the causative mutation in 266 HA patients from Emilia-Romagna region (Italy) and in all suspected carriers. Molecular analysis of F8 in 201 HA patients (152 index cases) was performed with a combination of several indirect and direct molecular approaches, such as long distance polymerase chain reaction, multiplex ligation-dependent probe amplification, denaturing high performance liquid chromatography and direct sequencing. The analysis revealed 78 different mutations, 23 of which were novel, not having been reported in national or international databases. The detection rate was 100%, 86% and 89% in patients with severe, moderate and mild HA, respectively. The information provided by this registry will be helpful for monitoring the treatment of HA patients in Emilia-Romagna and also for reliable genetic counselling of affected families in the future.

Mortality and causes of death in Italian persons with haemophilia, 1990-2007.

Although a number of studies have analysed so far the causes of death and the life expectancy in haemophilic populations, no investigations have been conducted among Italian haemophilia centres. Thus, the aim of this study was to investigate mortality, causes of deaths, life expectancy and co-morbidities in Italian persons with haemophilia (PWH). Data pertaining to a total of 443 PWH who died between 1980 and 2007 were retrospectively collected in the 30 centres who are members of the Italian Association of Haemophilia Centres that chose to participate. The mortality rate ratio standardized to the male Italian population (SMR) was reduced during the periods 1990-1999 and 2000-2007 such that during the latter, death rate overlapped that of the general population (SMR 1990-1999: 1.98 95% CI 1.54-2.51; SMR 2000-2007: 1.08 95% CI 0.83-1.40). Similarly, life expectancy in the whole haemophilic population increased in the same period (71.2 years in 2000-2007 vs. 64.0 in 1990-1999), approaching that of the general male population. While human immunodeficiency virus infection was the main cause of death (45%), 13% of deaths were caused by hepatitis C-associated complications. The results of this retrospective study show that in Italian PWH improvements in the quality of treatment and global medical care provided by specialized haemophilia centres resulted in a significantly increased life expectancy.

A web-based registry of inherited bleeding disorders in the region of Emilia-Romagna: results at three and a half years.

A Registry of inherited bleeding disorders was set up in the Region of Emilia-Romagna (RER) to collect information about these diseases and to improve the quality of care. From January 2003, the eight Haemophilia Centres (HC) in the RER began to use computerized clinical records; every 6 months, they send data to Parma Hospital to be processed and published in a website (http://www.registroemofiliarer.it). Great efforts are made to ensure high quality of data. Results of general interest are included in a free 'public area' and more sensitive data in a 'reserved area' (open only to HC and to health authorities). A total of 610 individuals are included: 249 haemophilia A (HA), 63 haemophilia B (HB), 173 von Willebrand's disease, 69 rare bleeding disorders, seven platelet disorders and 49 haemophilia carriers; 131 were genotyped, 188 were tested for inhibitors (16 affected). The most frequent bleeding was haemarthrosis. The joint score (evaluated in 104 haemophiliacs) was higher in severe HA. There were 22 HIV-positive and 182 hepatitis C virus-positive patients (21% have chronic hepatitis, two hepatocellular carcinoma). In 2005, two patients received primary prophylaxis, 47 secondary prophylaxis, four children were on immune-tolerance induction. From 2003 to 2005 the use of recombinant products was greatly increased and the majority of patients received them. The mean clotting factor consumption for prophylaxis was higher than on-demand treatment. The main features of registry are to collect high quality and comprehensive data of all patients followed by HC, to improve quality of care and it's availability on the web.

Prothrombin complex concentrates for urgent anticoagulation reversal in patients with intracranial haemorrhage.

BACKGROUND: Intracranial haemorrhage (ICH) is a serious and potentially fatal complication of oral anticoagulant therapy (OAT). Prothrombin complex concentrates (PCCs) produce a rapid and effective reversal of OAT effects, but little evidence exists on their efficacy and safety in the management of ICH in patients on OAT. AIM: To evaluate the efficacy and safety of PCCs for the rapid reversal of OAT in patients with ICH. METHODS: Patients suffering from acute ICH while receiving OAT were eligible for this prospective cohort study if their international normalized ratio (INR) was > or = 2.0. Stratified 35-50 IU kg(-1) PCC doses were infused based on initial INR. RESULTS: A total of 92 patients (50 males; mean age 74 years, range 34-92 years) were included. The median INR at presentation was 3.3 (range 2-9). At 30 min after PCC administration the median INR was 1.4 (range 0.9-3.1), declining to < or = 1.5 in 75% of patients. The benefit of PCC was maintained for a long time, since in 98% of all post-infusion time points through 96 h the median INR remained < or = 1.5 (median 1.19; range 0.9-2.3). During hospitalization neither thrombotic complications nor significant adverse events were observed and 11 patients died (11.9%). None of the deaths was judged to be related to PCC administration. CONCLUSIONS: PCC administration is an effective, rapid and safe treatment for the urgent reversal of OAT in patients with ICH. Broader use of PCC in this clinical setting appears to be appropriate.

[The antiphospholipid syndrome during chronic lymphatic leukemia. An association with anti-factor VIII antibodies]. / Sindrome da anticorpi antifosfolipidi in corso di leucemia linfatica cronica. Associazione con autoanticorpi anti-fattore VIII.

Activated partial thromboplastin time may be prolonged as the result of either of two different autoimmune complications of chronic lymphocytic leukemia: the development of antiphospholipid antibodies, such as lupus anticoagulant or anticardiolipin antibodies, or anti-factor VIII inhibitors, such as acquired hemophilia A. In the rare simultaneous occurrence of both inhibitors, differential diagnosis of a prolonged activated partial thromboplastin time poses a number of problems during laboratory work-up, due to mutual interference of the commonly performed tests. Only careful clinical follow-up can disclose the significance of the laboratory findings. We report the case of concurrent antiphospholipid antibodies (lupus anticoagulant positivity, anticardiolipin antibodies; IgM 3880 MPL/mL and IgG 265 GPL/mL) and anti-factor VIII antibodies (46.8 Bethesda Units) in a patient with chronic B-cell lymphocytic leukemia who had prolonged activated partial thromboplastin time (78.8 s). The relationship between lymphoproliferative and antiphospholipid syndrome, laboratory work-up in the case of the association of antiphospholipid and anti-factor VIII antibodies, and related problems that occur during clinical management of the patient are also discussed.

[Systemic mastocytosis: a review of the literature and of the cases in Reggio Emilia from 1986 to 1994]. / La mastocitosi sistemica: revisione della letteratura e della casistica di Reggio Emilia dal 1986 al 1994.

Systemic mastocytosis is a rare pathology that can affect most systems of the human organism. Although diagnosis is often fortuitous and prognosis good in a very high percentage of cases, it can sometimes present in extremely severe and occasionally fatal forms. With the aid of the available literature, we discuss the most recent classifications, clinical features and diagnostic and therapeutic approaches to this disease. We then do an epidemiological review of the cases reported in Reggio Emilia over the past 9 years. Due to the lack of symptoms of this pathology, its reported incidence of about 0.3 new cases per 100,000 inhabitants per year is obviously underestimated. Drug therapy is purely symptomatic and does not affect its clinical evolution.

Serum selenium concentrations in patients with newly diagnosed lymphoid malignancies.

BACKGROUND: Increased mortality from lymphoid malignancies following exposure to environmental selenium has recently been reported. Moreover, conflicting results have been found in investigations examining the relationship between serum concentrations of selenium and some clinical features of malignant lymphoproliferative diseases. METHODS: Serum concentrations of selenium were analyzed by atomic absorption spectrometry in fifty-nine patients with newly diagnosed chronic lymphoid malignancies and in forty control subjects. RESULTS: Selenium concentrations were significantly lower in patients than in control subjects. However, when only patients with localized disease were compared to controls, no significant difference in serum selenium concentrations was observed. Clinical stage was inversely associated with selenium levels. High-grade non-Hodgkin's lymphoma was characterized by lower selenium levels than low-grade and intermediate-grade disease. Selenium levels were positively associated with albumin and hemoglobin, and inversely correlated with serum concentrations of beta 2-microglobulin and with erythrocyte sedimentation rate. CONCLUSIONS: The findings of this study do not suggest that a high selenium intake represents a risk factor for malignant lymphoproliferative diseases, but limitations of the investigation hamper evaluation of the results. The possible utility of determining serum concentrations of selenium in the clinical evaluation of patients with lymphoid malignancies merits examination in larger studies.

Cost/benefit relationship in 46 cases of myelodysplastic syndrome (RAEB and RAEB-T): a quality assurance analysis in clinical haematology.

BACKGROUND: Myelodysplastic Syndromes (MDS) are common and severe haematologic diseases. Clinicians still have no standard therapies surely able to obtain a better result in the different MDS (RAEB and RAEB-t in particular). Moreover, the costs due to the specific drug, to its clinical complications and to the days spent in hospital can be influenced by the chosen therapeutic regime. There are no published studies regarding a cost-benefit relationship of the different drugs commonly used today for MDS patients. PATIENTS AND METHODS: This retrospective study refers to 46 patients with RAEB or RAEB-t, followed in our Division from 1986 up to December 1992. The authors reviewed specific therapies (ARA-C vs other drugs, in particular), the costs supported by the National Health Service and the effects of the same therapies in terms of survival. The results have been worked out by statistical analysis. RESULTS: Some interesting data are presented in this study: chemotherapy does not improve survival and causes higher costs; the length of stay in hospital is not influenced by using chemotherapy or not; RAEB patients cause as high costs as RAEB-t ones. CONCLUSIONS: Although the study regards only a limited number of cases, it shows that the therapeutic efforts made so far in order to improve MDS patients' prognosis cannot be considered satisfying and their social cost is very high. Our report can be a useful starting-point for a QA analysis concerning the rational use of new drugs such as growth factors and interleukins.

Ratios between the abundance of messenger RNA and the corresponding protein of two growth-related genes, c-myc and vimentin, in leukemia blast cells.

The abundance of the mRNAs of two growth-related genes, vimentin and c-myc, and that of the corresponding proteins have been studied in unstimulated and phytohemagglutinin-stimulated lymphocytes as well as in 18 populations of leukemic blast cells. The quantitative assay was carried out by densitometric scanning of Northern and Western blots. In normal lymphocytes the mRNA and the protein of both genes were almost undetectable. The phytohemagglutinin stimulation led to a sharp increase of the mRNA and the proteins of vimentin and c-myc. The increase was followed by a progressive fall of the gene products. The rate of decrease of the two mRNAs was similar to that of the corresponding proteins. In some leukemic populations very similar amounts of the vimentin protein were accompanied by amounts of the mRNA differing at least 25 times. Not unlikely, very similar amounts of p62c-myc corresponded to mRNA abundances differing at least 16 times. The coordinated biogenesis of both messenger RNAs and proteins, which occurs in mitogen-stimulated lymphocytes, is substituted, in approximately 30% of the leukemic blast cell populations, by molecular events leading to the accumulation of an excess of mRNA.