A recent view about encephalomyocarditis virus circulating in compartmentalised animal population in Northern Italy.

Encephalomyocarditis virus (Picornaviridae, Cardiovirus A) is the causative agent of the homonymous disease, which may induce myocarditis, encephalitis and reproductive disorders in various mammals, especially in swine. Despite the disease occurred endemically in pig farms since 1997, the recent increase of death experimented in Northern Italy prompted to furtherly investigate the evolution of the virus and the actual spread of the infection. Italian EMC viruses, collected between 2013 and 2019, showed an overall antigenic stability. The in-house ELISA Monoclonal Antibodies based, able to reveal changes in seven different antigenic sites, showed only sporadic and occasional mutations in considered samples and the subsequent phylogenetic analysis confirmed antigenic panel's remarks. All the isolates could be classified within a unique lineage, which comprise other European strains and confirm that the viruses currently circulating in Italy developed from a unique common ancestor. Despite the demonstrated stability of virus, some putative newly emerged variants were detected through antigenic profile analysis and phylogenesis. Finally, the serosurvey proved that spread of EMCV is greater than the diffusion of fatal infections would suggest, due to subclinical circulation of EMCV. It demonstrated an increase in the proportion of seropositive farms, if compared with previous data with no remarkable differences between farms with and without clinical evidence of disease.

Characterization of the O/ME-SA/Ind-2001d foot-and-mouth disease virus epidemic recorded in the Maghreb during 2014-2015.

The O/ME-SA/Ind-2001d has been the main foot-and-mouth disease virus (FMDV) lineage responsible for FMD epidemics outside the Indian subcontinent from 2013 to 2017. In 2014, outbreaks caused by this FMDV lineage were reported in Maghreb, where it was initially detected in Algeria and Tunisia and later in Morocco. This was the first incursion of an FMDV type O of exotic origin in the Maghreb region after 14 years of absence. In this study, we report analyses of both VP1 and whole-genome sequences (WGSs) generated from 22 isolates collected in Algeria and Tunisia between 2014 and 2015. All the WGSs analysed showed a minimum pairwise identity of 98.9% at the nucleotide level and 99% at the amino acid level (FMDV coding region). All Tunisian sequences shared a single putative common ancestor closely related to FMDV strains circulating in Libya during 2013. Whereas sequences from Algeria suggest the country experienced two virus introductions. The first introduction is represented by strains circulating in 2014 which are closely related to those from Tunisia, the second one, of which the origin is more uncertain, includes strains collected in Algeria in 2015 that gave origin to the 2015 outbreak reported in Morocco. Overall, our results demonstrated that a unique introduction of O/Ind-2001d FMDV occurred in Maghreb through Tunisia presumably in 2014, and from then the virus spread into Algeria and later into Morocco.

Detection of Bovine Antibodies against a Conserved Capsid Epitope as the Basis of a Novel Universal Serological Test for Foot-and-Mouth Disease.

Diagnostic tests for foot-and-mouth disease (FMD) include the detection of antibodies against either the viral nonstructural proteins or the capsid. The detection of antibodies against the structural proteins (SP) of the capsid can be used to monitor seroconversion in both infected and vaccinated animals. However, SP tests need to be tailored to the individual FMD virus (FMDV) serotype and their sensitivity may be affected by antigenic variability within each serotype and mismatching between test reagents. As a consequence, FMD reference laboratories are required to maintain multiple type-specific SP assays and reagents. A universal SP test would simplify frontline diagnostics and facilitate large-scale serological surveillance and postvaccination monitoring. In this study, a highly conserved region in the N terminus of FMDV capsid protein VP2 (VP2N) was characterized using a panel of intertype-reactive monoclonal antibodies. This revealed a universal epitope in VP2N which could be used as a peptide antigen to detect FMDV-specific antibodies against all types of the virus. A VP2-peptide enzyme-linked immunosorbent assay (VP2-ELISA) was optimized using experimental and reference antisera from immunized, convalescent, and naïve animals (n = 172). The VP2-ELISA is universal and simple and provided sensitive (99%) and specific (93%) detection of antibodies to all FMDV strains used in this study. We anticipate that this SP test could have utility for serosurveillance during virus incursions in FMD-free countries and as an additional screening tool to assess FMD virus circulation in countries where the disease is endemic.

Development and validation of a foot-and-mouth disease virus SAT serotype-specific 3ABC assay to differentiate infected from vaccinated animals.

The effective control of foot-and-mouth disease (FMD) requires sensitive, specific and rapid diagnostic tools. However, the control and eradication of FMD in Africa is complicated by, among other factors, the existence of five of the seven FMD virus (FMDV) serotypes, including the SAT-serotypes 1, 2 and 3 that are genetically and antigenically the most variable FMDV serotypes. A key diagnostic assay to enable a country to re-gain its FMD-free status and for FMD surveillance, is the 3ABC or the non-structural protein (NSP) enzyme-linked immunosorbent assay (ELISA). Although many kits are available to detect 3ABC antibodies, none has been developed specifically for the variable SAT serotypes. This study designed a SAT-specific NSP ELISA and determined whether this assay could better detect NSP-specific antibodies from FMDV SAT-infected livestock. The assay's performance was compared to validated NSP assays (PrioCheck®-NSP and IZSLER-NSP), using panels of field and experimental sera, vaccinated and/or infected with FMDV SAT1, SAT2 or SAT3. The sensitivity () of the SAT-NSP was estimated as 76% (70%, 81%) whereas the specificity was 96% (95%, 98%) at a 95% confidence interval. The sensitivity and specificity were comparable to the commercial NSP assays, PrioCheck®-NSP (82% and 99%, respectively) and IZSLER-NSP (78% and 98%, respectively). Good correlations were observed for all three assays.

Study on seroprevalence and serotyping of foot and mouth disease in Chad.

Foot and mouth disease (FMD) is a highly contagious viral disease that affects all Artiodactyla. Seven immunologically distinct serotypes of FMD virus (FMDV) exist. In Chad, although FMD is included in the list of diseases monitored by the Chadian Animal Disease Surveillance Network (REPIMAT), the epidemiological situation remains unclear. A serological survey was conducted in the cattle population in eight of the nine administrative regions of the country (those regions with the highest cattle densities), to evaluate the prevalence and serotypes of circulating FMDV.A total of 796 sera from randomly selected cattle were analysed at the World Organisation for Animal Health/Food and Agriculture Organization of the United Nations FMD Reference Laboratory at the Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna(IZSLER), in Brescia (Italy). An enzyme-linked immunosorbent assay (ELISA), called 3ABC ELISA, was used to detect antibodies against non-structural proteins (NSPs), as well as a series of six competitive ELISAs to detect and serotype antibodies against the structural proteins of FMDV serotypes O, A, SAT 1, SAT 2, Asia 1 and C. Based on the detection of anti-NSP antibodies, the animal-level seroprevalence was 35.6%(95% confidence interval [CI]: 32.2-38.9) and the herd-level seroprevalence was 62.3% (95%CI: 53.0-71.5). FMD was present in all livestock administrative divisions surveyed, with a higher prevalence in southern regions, which are characterised by higher rainfall and humidity and more important transboundary animal movements. Cattle aged more than four years had a higher seroprevalence, which may be due to repeated exposure. Semi-sedentary farming and transhumance were also risk factors. Antibodies against serotypes A, O, SAT 1 and SAT 2 were detected.

La fièvre aphteuse est une maladie virale extrêmement contagieuse qui affecte l'ensemble des artiodactyles. Sept sérotypes du virus de la fièvre aphteuse ont été répertoriés, qui sont distincts au plan immunologique. Au Tchad, bien que la fièvre aphteuse figure sur la liste des maladies visées par le Réseau d'épidémiosurveillance des maladies animales du Tchad (REPIMAT), la situation épidémiologique demeure mal connue. Une enquête sérologique a été réalisée dans la population bovine de huit régions administratives sur les neuf que compte le pays (afin de couvrir les régions où la densité de la population bovine est la plus élevée), dans le but d'évaluer la prévalence du virus de la fièvre aphteuse et de caractériser les sérotypes présents. Au total, 796 sérums prélevés sur des bovins sélectionnés de manière aléatoire ont été analysés au Laboratoire de référence pour la fièvre aphteuse de l'Organisation mondiale de la santé animale/Organisation des Nations Unies pour l'alimentation et l'agriculture à l'Istituto Zooprofilattico Sperimentale dellaLombardia e dell'Emilia Romagna (IZSLER) de Brescia (Italie). Les anticorps dirigés contre les protéines non structurales ont été détectés au moyen d'une épreuve immuno-enzymatique 3ABC (ELISA 3ABC) tandis qu'une série de six ELISA de compétition a permis de détecter et de caractériser les anticorps spécifiques des protéines structurales des sérotypes O, A, SAT 1, SAT 2, Asia 1 et C du virus de la fièvre aphteuse. D'après les résultats de la détection d'anticorps dirigés contre les protéines non structurales, la prévalence sérologique à l'échelle individuelle était de35,6 % (avec un intervalle de confiance [IC] à 95 % de 32,2 à 38,9) tandis que la prévalence à l'échelle des troupeaux s'élevait à 62,3 % (IC à 95 % de 53,0 à 71,5).La fièvre aphteuse était présente dans chacune des divisions administratives étudiées, avec une prévalence plus élevée dans les régions méridionales, qui se caractérisent par des précipitations et une hygrométrie plus fortes et par l'importance des mouvements transfrontaliers d'animaux. La prévalence sérologique était plus élevée chez les bovins âgés de plus de quatre ans, ce qui s'explique probablement par un nombre répété d'expositions. Le rôle de l'élevage semi-sédentaire et de la transhumance en tant que facteurs de risque a été misen lumière. Les anticorps détectés étaient dirigés contre les sérotypes A, O, SAT 1et SAT 2.

La fiebre aftosa es una patología vírica muy contagiosa que afecta a todos los artiodáctilos. Existen siete serotipos inmunológicamente diferenciados del virus que la causa. En el Chad, aunque la fiebre aftosa figura en la lista de enfermedades sometidas a vigilancia por la Red Chadiana de Vigilancia Zoosanitaria (REPIMAT), la situación epidemiológica de la enfermedad sigue rodeada de incertidumbre. Los autores describen un estudio serológico realizado en la población vacuna de ocho de las nueve regiones administrativas del país (las que presentan la mayor densidad de ganado vacuno) con objeto de determinar la prevalencia y los serotipos del virus de la fiebre aftosa circulante. Tras seleccionar aleatoriamente un total de 796 cabezas de ganado y obtener de ellas muestras de suero, estas fueron analizadas en el Istituto Zooprofilattico Sperimentale della Lombardia e dell'EmiliaRomagna (IZSLER) de Brescia (Italia), que es el Laboratorio de Referencia de la Organización Mundial de Sanidad Animal y la Organización de las Naciones Unidas para la Alimentación y la Agricultura para la fiebre aftosa. Para detectar anticuerpos dirigidos específicamente contra proteínas no estructurales se empleó un ensayo inmunoenzimático (ELISA) denominado ELISA 3ABC, a lo que se agregó una serie de seis técnicas ELISA de competición concebidas para detectar y tipificar anticuerpos dirigidos contra las proteínas estructurales de los serotipos O, A,SAT 1, SAT 2, Asia 1 y C del virus de la fiebre aftosa. A tenor de los niveles detectados de anticuerpos contra proteínas no estructurales, la seroprevalencia individual era de un 35,6% (intervalo de confianza [IC] al 95%:32,2­38,9) y la seroprevalencia de rebaño era de un 62,3% (IC 95%: 53,0­71,5).La fiebre aftosa, presente en todas las divisiones administrativas ganaderas estudiadas, alcanzaba sus máximos niveles de prevalencia en las regiones meridionales, caracterizadas por tasas de pluviosidad y humedad más altas y por un mayor volumen de movimientos transfronterizos de animales. La seroprevalencia era más elevada en los ejemplares de más de cuatro años de edad, hecho que puede deberse a exposiciones reiteradas. La producción ganadera en régimen semisedentario y la trashumancia eran también factores de riesgo. Se detectaron anticuerpos contra los serotipos A, O, SAT 1 y SAT 2.

Sero-prevalence and epidemiology of peste des petits ruminants in Libya.

We conducted a cross-sectional study during 2013 to quantify the serological prevalence of peste des petits ruminants (PPR) infection and to investigate host factors associated with PPR infection in small ruminants in Libya. A two-stage sampling design was carried out. A total number of 148 flocks owning at least 100 heads each were randomly selected. Sixteen to forty-eight samples were collected from each selected flock. A total number of 3,508 serum samples from unvaccinated animals were collected and analysed at IZSLER Brescia, Italy, by using competitive ELISA, IDvet innovative diagnostics (IDvet 310, France). The overall serological prevalence among SR was 33% (95% CI: 31.4-34.5). Significant differences between the prevalence in the geographical branches were observed. The lowest prevalence level was observed in Zawiyah branch (16.1%), whereas the highest value was obtained for the Sabha branch (56.8%). Considering the age, a serological prevalence of 24.7%, 31.5% and 42.1% was observed in SR <1 year, between 1 and 2 years and more than 2 years, respectively. Statistically significant differences (p < .001) in the sero-prevalence levels were also observed between the age groups. Our findings suggest that the southern part of Libya could be more exposed to the infections coming from the neighbouring countries and this should be better investigated to correctly identify wherever specific entry points can be considered at higher risk than others. The results also confirmed the endemic status of PPR in Libya, with a constant exposure to the infection of the animals during their life. In the framework of the global strategy for control and eradication of PPR, our results, even if obtained by a preliminary study, can contribute to the assessment of the epidemiological situation of PPR in Libya as required by the Stage 1 of the plan.

Outbreaks of Foot-and-Mouth Disease in Libya and Saudi Arabia During 2013 Due to an Exotic O/ME-SA/Ind-2001 Lineage Virus.

Foot-and-mouth disease viruses are often restricted to specific geographical regions and spread to new areas may lead to significant epidemics. Phylogenetic analysis of sequences of the VP1 genome region of recent outbreak viruses from Libya and Saudi Arabia has revealed a lineage, O-Ind-2001, normally found in the Indian subcontinent. This paper describes the characterization of field viruses collected from these cases and provides information about a new real-time RT-PCR assay that can be used to detect viruses from this lineage and discriminate them from other endemic FMD viruses that are co-circulating in North Africa and western Eurasia.

Pulmonary embolism severity index accurately predicts long-term mortality rate in patients hospitalized for acute pulmonary embolism.

BACKGROUND: The Pulmonary Embolism (PE) Severity Index (PESI) is a clinical prognostic rule that accurately classifies PE patients into five risk classes with increasing mortality. PESI score has been validated in studies with a relatively short-term follow-up and its accuracy in predicting long-term prognosis has never been established. METHODS: Consecutive patients admitted to the tertiary care hospital of Varese (Italy) with an objectively diagnosed PE between January 2005 and December 2009 were retrospectively included. Information on clinical presentation, diagnostic work-up, risk factors, treatment and mortality during a 1-year follow-up was collected. RESULTS: Five hundred and thirty-eight patients were enrolled in this study. The mean age was 70.6 years (± SD 15.2), 44.4% of patients were male, and 27.9% had known cancer. One-year follow-up was available for 96.1% of patients. The overall mortality rate was 23.2% at 3 months, 30.2% at 6 months and 37.1% at 12 months. The discriminatory power of the PESI score to predict long-term mortality, expressed as the area under the ROC curve, was 0.77 (95%CI, 0.72-0.81) at 3 months, 0.77 (95%CI, 0.73-0.81) at 6 months and 0.79 (95%CI, 0.75-0.82) at 12 months. The PESI score confirmed its accurate prediction in patients without cancer. Simplified PESI had a similar overall accuracy to the original PESI at 3 and 6 months, but this was significantly lower at 1 year. CONCLUSIONS: The results of this study suggest that PESI score may also be an accurate tool to define the 6-month and 1-year mortality rates in PE patients.

Development and characterization of a bovine serum evaluation panel as a standard for immunoassays based on detection of antibodies against foot-and-mouth disease viral non-capsid proteins.

The widespread perception of the effectiveness of applying tests based on the detection of antibodies against foot-and-mouth disease (FMD) viral non-capsid proteins (NCPs) to assess virus circulation irrespective of vaccination triggered the demand for international standards to evaluate the comparative performance of the upcoming assays against the OIE Index test developed at the Pan American Foot-and-Mouth Disease Center, PAHO/WHO. To this end, a panel was developed composed of 34 cattle sera from animals with an unambiguous exposed/infected status, covering serotypes O, A and C, obtained either under experimental conditions or from the field in regions with different epidemiological situations. Reference values in the Index test and their reproducibility in other laboratories, data on stability as well as results in four other commercial kits and one in house test were obtained. The characteristics of the panel which comprise adequate preparation following international guidelines, a broad range of antibody reactivity, proper stability and the ability to assess comparative diagnostic sensitivity, make it suitable as a reference standard to evaluate if tests equivalent to the OIE Index method are used in support of FMD control programs and by trading partners, and also whether they maintain their standards of diagnostic performance.

Comparative evaluation of six ELISAs for the detection of antibodies to the non-structural proteins of foot-and-mouth disease virus.

To validate the use of serology in substantiating freedom from infection after foot-and-mouth disease (FMD) outbreaks have been controlled by measures that include vaccination, 3551 sera were tested with six assays that detect antibodies to the non-structural proteins of FMD virus. The sera came from naïve, vaccinated, infected and vaccinated-and-infected animals; two-thirds from cattle, the remainder from sheep and pigs. The assays were covariant for sensitivity, but not necessarily for specificity. A commercial kit from Cedi-diagnostics and an in-house assay from IZS-Brescia were comparable to the NCPanaftosa-screening index method described in the Diagnostic Manual of the World Animal Health Organisation. Using these three tests the specificity and sensitivity for the detection of carriers in vaccinated cattle approaches or exceeds 99% and 90%, respectively.

Differentiation of foot-and-mouth disease virus infected animals from vaccinated animals using a blocking ELISA based on baculovirus expressed FMDV 3ABC antigen and a 3ABC monoclonal antibody.

A blocking ELISA that differentiated foot-and-mouth disease virus (FMDV) infected animals from vaccinated animals was developed which uses baculovirus expressed FMDV 3ABC non-structural protein as antigen and monoclonal antibody against FMDV 3ABC non-structural protein as capture and detector antibody. Sera from naive, vaccinated and infected cattle, sheep and pigs were examined. The specificity of the test was high. Non-specific reactions observed in particular in sera of cattle and sheep could be removed by filtration and inactivation. Positive reactions were obtained for sera from cattle infected with all seven serotypes of FMDV. The test detected antibodies from days 7 or 9 following experimental infection of non-vaccinated cattle and sheep, and in cattle strong positive reactions persisted for up to 395 days after infection. In vaccinated cattle that became carriers after challenge with homologous FMDV, positive reactions were obtained in all but one case. In some of these cattle the antibody response was detected late in comparison to the non-vaccinated infected cattle. The test gave results that compared favourably with two commercial ELISA's when used to test sera from cattle, pigs and sheep collected after experimental or natural infection. The blocking ELISA based on recombinant FMDV 3ABC antigen and a monoclonal antibody to 3ABC is a promising tool for FMD control and eradication campaigns, where vaccination has been carried out.

Differentiating infection from vaccination in foot-and-mouth-disease: evaluation of an ELISA based on recombinant 3ABC.

Recent devastating outbreaks of foot-and-mouth disease (FMD) in Europe have reopened the discussion about the adequacy of the non-vaccination strategy implemented by the EU in 1991. Here we describe the evaluation of a new commercially available test kit for the discrimination between vaccination and infection. The test is based on the detection of antibodies against the recombinant non-structural (NS) protein 3ABC. In contrast to immunization with vaccines free of 3ABC, these antibodies are elicited as a consequence of infection. Testing more than 3600 negative sera from several countries revealed a specificity of > 99% for bovine, ovine, and porcine samples. Antibodies specific for 3ABC can be detected as soon as 10 days post-infection. As compared with the occurrence of antibodies against structural proteins of FMDV, anti-3ABC antibodies can be detected 5-10 days later, depending on the species. No anti-3ABC antibodies were detected in sera from vaccination experiments or in field sera from vaccinated animals. However, anti-3ABC antibodies can be detected in vaccinated animals upon challenge. These results provide evidence that this test can facilitate the use of vaccines in new strategies against FMD.

Homozygosity for angiotensinogen 235T variant increases the risk of myocardial infarction in patients with multi-vessel coronary artery disease.

OBJECTIVE: Molecular variants of the angiotensinogen (AGT) and the angiotensin II type 1 receptor (ATR) genes have been associated with the risk of coronary artery disease (CAD) and myocardial infarction (MI), but data so far available are conflicting. The primary object of the paper is to verify this possible association by a rigorous, angiographically controlled study in a large sample of patients with or without multi-vessel CAD. DESIGN: We designed a large case-control study in Italian patients candidates for coronary artery bypass grafting, with angiographically documented multi-vessel CAD, compared to subjects with angiographically documented normal coronary arteries. METHODS AND RESULTS: AGT M235T and ATR A1166C gene polymorphisms were analysed in 699 subjects; 454 patients were candidates for coronary artery bypass grafting, having angiographically documented (mainly multi-vessel) CAD. An appropriate documentation of previous MI was obtained from 404/454 (89%, 247 with and 157 without MI). Subjects (n = 245) with angiographically documented normal coronary arteries, were included as control group (CAD-free group). CAD patients had a substantial burden of conventional risk factors as compared with controls free of coronary atherosclerosis. Age, gender, smoking habit and number of stenosed vessels were the only differences between patients with or without previous myocardial infarction, who were similarly exposed to the other conventional risk factors (including hypertension). AGT M235T and ATR A1166C allele and genotype frequencies were similar between CAD and CAD-free patients. In the CAD group, AGT 235T allele was found more frequently in subjects with a previous myocardial infarction (0.494 versus 0.414; P < or = 0.05). By logistic regression, homozygosity for AGT 235T variant appeared to confer 1.9-fold increased risk for MI in both the univariate and the multivariate (adjusted for age, gender, smoking habit and number of stenosed vessels) model. CONCLUSIONS: AGT 235 T homozygous patients with multivessel CAD have an increased risk of myocardial infarction as compared with subjects with clinically similar phenotype but different genotype.

Genetic polymorphisms of the renin-angiotensin system and atheromatous renal artery stenosis.

Genes that influence the renin-angiotensin system have been investigated in recent years as potential etiologic candidates of cardiovascular and renal diseases. In atheromatous renal artery stenosis (RAS), a condition characterized by persistent activation of the renin-angiotensin system, the study of these genes may be of particular relevance. We evaluated angiotensin-converting enzyme (ACE) insertion/deletion, angiotensinogen (AGT) M235T, and angiotensin II receptor (ATR) A1166C polymorphisms in relation to the occurrence of RAS. We studied 58 patients with angiographically documented RAS; 102 normotensive subjects with normal coronary arteries and no history or clinical or instrumental evidence of atherosclerosis in other vascular districts were considered the control group. Patients had a significantly higher D allele frequency (0.70 versus 0.55; chi(2) 6.88, P=0.01; odds ratio [OR] 1. 9, 95% CI 1.17 to 3.07) than did the control population; 48.3% of patients were homozygous for DD (chi(2) 6.62, P<0.05; OR 2.04, 95% CI 1.05 to 3.95); and only 8.6% carried the II genotype (OR 0.34, 95% CI 0.19 to 1.47). No significant association was found for AGT M235T and ATR A1166C. Our results suggest a predisposing role for ACE genetic polymorphism in the development and progression of atheromatous RAS.

[Is mammography useful in the detection of breast cancer in women 35 years of age or younger?]. / E utile la mammografia nel cancro della mammella nelle donne con età pari o inferiore a 35 anni?

Breast cancer in women 35 years old or younger is unusual. It accounts for 1-3.6% of all breast cancers but is the leading cause of cancer mortality in women 15-35 years old. The diagnostic delay, with T2 or more advanced cancer at clinical presentation, is due to the patient's age and the opinion of low mammographic reliability for cancer diagnosis in this age group. To assess the usefulness of mammography in breast cancer patients aged 35 years or younger, we reviewed the clinical, mammographic and histologic data of 65 cancers collected in 7 breast diagnosis and counseling centers in Lombardy. Fifty-three patients (81.5%) were referred for a palpable breast mass, which was a T2 or more advanced cancer in 23 cases. Mammography showed malignant patterns (spiculated opacities, clusters of microcalcifications, casting, branching and ductal type calcifications) in 31 patients (47.7%). Mammography was not definitive but correctly suggested further examinations in 30 women and it had only 4 false negatives. Ultrasonography performed in 43 patients was negative in 3 (7%), pathologic and pathognomonic for cancer in 27 (62.8%) and pathologic but not indicative of malignancy in 13 (20.2%). The cytologic or histologic diagnosis of breast cancer was made under US guidance in 24 cases. In women aged 35 years or younger mammography was effective in identifying breast cancers; US and fine-needle aspiration biopsy (FNAB) complete mammography. We believe that mammography can be a valuable screening tool in young women at high risk for breast cancer because of family history.

A further step in the evolution of rabbit hemorrhagic disease virus: the appearance of the first consistent antigenic variant.

Rabbit hemorrhagic disease virus (RHDV) is a noncultivable calicivirus that infects rabbits (Oryctolagus cuniculus) and causes epidemics of an acute fatal hepatitis. In 1997 we identified two RHDV isolates from geographically distant Italian regions, which differed antigenically from the reference strain RHDV.Bs89. In fact, they were not reactive with mAb 1H8, that is able to protect rabbits from RHD and showed a low reactivity with the rabbit convalescent serum raised against RHDV.Bs89. Experimental infection of rabbits with either RHDV isolates confirmed their high pathogenicity and their peculiar antigenic profile; nevertheless, rabbits vaccinated with the current vaccine were protected against challenge infection with these isolates. Sequence comparison definitely demonstrated that the two isolates originated from the same RHDV variant and that the similarity of their structural protein (VP60) sequences with the RHDV.Bs89 is equal to 93%. This variant was named RHDVa since shows consistent genetic and antigenic differences from the wild-type RHDV. In particular, 44% of amino acid substitutions in RHDVa VP60 were located between amino acids 344 and 370, where the similarity with RHDV.Bs89 drops to 70%, suggesting that this region probably contains the epitope recognized by mAb 1H8. In addition, this paper presents preliminary data concerning the amino acids of VP60 involved in the hemagglutination site of the virus.