New Insights Into the Genetic Variability of Fasciola Hepatica (Trematoda) in Algeria and Relationships with Other Geographic Regions Revealed By Mitochondrial DNA.

This study aims to investigate the level of genetic variability of Fasciola hepatica flukes isolated from cattle in Algeria and to determine the phylogenetic and phylogeographic relationships with sequences isolated worldwide. Mitochondrial (Cytochrome c Oxidase subunit I gene - COI) and nuclear markers (Internal Transcribed Spacers of nuclear ribosomal DNA - ITS) for 24 F. hepatica flukes isolated from 12 cattle in North Algeria were characterised. Only two haplotypes were obtained for the COI gene, resulting in a low level of genetic variation. The analysis of variation among the COI sequences isolated from around the world did not show high levels of genetic divergence, and the phylogenetic analysis revealed a genetic similarity among F. hepatica isolates from different areas of the world. The analysis of the ITS region showed a low level of variability, which prevented obtaining informative phylogenetic and phylogeographic results. The present study also revealed that specimens of F. hepatica are genetically similar in different hosts, indicating that the genetic structure among populations of this parasite is not influenced by the host species. The low levels of genetic variation for COI and ITS regions among fluke isolates from all continents are consistent with a common origin for the flukes' worldwide distribution.

Role of the "rooming-in" on efficacy of universal neonatal hearing screening programmes.

Sensorineural hearing loss is one of the most common congenital abnormalities in infants and it affects approximately one to two neonates in every 1000 births. Early identification of hearing loss in the newborn is the first step for a successful rehabilitation programme. The introduction of Otoacoustic Emission responses as a useful tool in hearing screening programmes, allowed the identification of hearing loss in the well-baby nursery and in targeted populations of the neonatal intensive care unit. Recently, a new concept of breastfeeding during hospitalization after birth has been developed. Indeed, the so-called "rooming-in" allows a mother to stay with her child in the same room, located in the nursery. This new trend has been developed to avoid any psychological adverse consequences of birth on the child-mother relationship. To enhance how "rooming-in" could affect the Universal Neonatal Hearing Screening (UNHS) programmes, an analysis has been made of the data coming from two maternity wards using different breastfeeding protocols. Data obtained demonstrate a worse performance on obtaining essential benchmark in the UNHS in the maternity ward where rooming-in is adopted (60% of newborns tested). UNHS programme efficacy could be affected by the wider adoption of the "rooming-in" regimen in the maternity wards and early detection of hearing loss revealed by UNHS could be vanished by dispersion of patients. In fact, more data are necessary to evaluate the impact of rooming, even though our data show a worsening in the UNHS results.

Sardinian multiple sclerosis is associated with HLA-DR4: a serologic and molecular analysis.

HLA haplotypes in 45 unrelated Sardinian multiple sclerosis patients and in six multiplex families were defined, using both serologic and restriction fragment length polymorphism (RFLP) analysis. In unrelated MS patients, we found an association with HLA-DR4 (p less than 0.01, relative risk = 2.5) and DQw3 (p less than 0.04, relative risk = 2.2). Using a beta-DR cDNA probe, we observed no variation of the DR4 RFLP profile in sporadic or related MS patients compared with DR4-specific pattern in controls. Using a beta-DQ cDNA probe, we identified two DQw3 patterns (DQw3.1 and DQw3.2) with similar frequency in patients and in controls. No specific RFLPs were observed in association with different disease courses. The frequency of haplotype sharing in affected members of multiplex families was not different from that expected by chance. This study shows that Sardinian MS patients carry predominantly the HLA-DR4 allele, in contrast to the DR2 prevalence reported in Caucasian populations. The lack of association with HLA haplotypes in affected members of multiplex families may indicate that genetic factors outside the HLA system play a substantial role in families with MS.

Chronic liver disease in transfusion-dependent thalassaemia: hepatitis B virus marker studies.

The systematic screening of 253 children with transfusion-dependent homozygous beta-thalassaemia revealed a high incidence of hepatitis B virus markers. The highest frequencies of hepatitis B surface antigen (HBsAg) and antibody to hepatitis B core antigen (anti-HBc) were found in the group of patients with the smallest number of transfusions, while the highest frequency of antibody to hepatitis B surface antigen (anti-HBs) was detected in the patients who had had the largest number of transfusions. Follow-up of these patients showed (a) a high incidence of acute hepatitis B, which was mainly subclinical; (b) normal hepatitis B surface antigen clearance and normal antibody to hepatitis B surface development; and (c) a high frequency of increased transaminase values for over six months. In all the subjects with persistently high transaminase, histological examination revealed chronic persistent hepatitis or chronic active hepatitis. Apart from two cases of chronic active hepatitis with no B virus markers, and two cases of chronic persistent hepatitis with HBsAg and anti-HBc in the serum, all these subjects were anti-HBs positive but HGsAg and anti-HBc negative.

Antenatal diagnosis of thalassemia major in Sardinia.

In this report, we summarized our experience, carried out in Sardinia, with antenatal diagnosis in one thousand pregnancies in which the fetus was at risk for homozygous beta-thalassemia. In the majority of these cases, the thalassemia lesion segregating in the family was the nonsense mutation at the codon corresponding to amino-acid 39. At the outset (976 cases) we used globin chain synthesis analysis by column chromatography on fetal blood obtained by placental aspiration, and recently (24 cases) we employed the synthetic oligonucleotide method on amniocyte DNA. Apart from 126 pregnancies still in progress, in all the other cases the diagnosis has been confirmed. In the majority of the cases (99%), we obtained sufficient fetal blood for the analysis. The fetal mortality associated with placental aspiration was 6.1%. The biochemical analysis gave reliable results. We had two misdiagnoses (0.2%): one due to a nonglobin protein comigrating with the beta chains and the other for a misclassification of the type of thalassemia segregating in the family. The oligonucleotide method gave clear-cut results in all the cases tested. The method was sensitive enough to detect the mutation directly in the DNA isolated from 20-25 ml of amniotic fluid in 75% of the pregnancies tested. In one case, we successfully employed this method for the analysis of the DNA isolated from chorionic villi. The oligonucleotide method seems to be the best procedure for monitoring the pregnancies at risk for beta-thalassemia in places where one or a few beta-thalassemia lesions are prevalent.

A strategy for fragile-X carrier screening.

Fragile-X syndrome is due to an expression of CGG trinucleotide repeats in the 5' untranslated region of the FMR1 gene and it is the most common cause of heritable X-linked mental retardation. Until now, the disease and the carrier state were diagnosed by Southern blotting or PCR-based methods. Southern blotting is an expensive, time-consuming, and radioisotope-based method that cannot easily be used for routine screening of an at-risk population. Nonradioisotopic PCR methods do not identify full mutated alleles, nor do they discriminate between alleles in the normal range that differ only by one or two CGG repeats. Therefore, two normal alleles with only a small difference in size, cannot be differentiated after PCR in Metaphor agarose or acrylamide gels. To define the genotype, it is necessary to perform Southern blot analysis. In this paper, we present a new strategy which, because of its simplicity, can be applied to large-scale fragile-X carrier screening of at-risk females.

Temporal and spatial patterns of African swine fever in Sardinia.

Temporal patterns and spatial distribution of African swine fever (ASF) were studied through the analysis of routinely collected data in the ASF-endemic area of the Province of Nuoro, Sardinia. During 1993-1996, ASF outbreaks were reported from 45 out of the 82 municipalities of the study area. Overall farm-level incidence rate (IR) was 1.3 outbreaks per 100 farms-year. ASF peaked in 1995 (IR = 1.8) and declined in 1996 (IR = 0.82). Significant (P < 0.05) spring peaks of ASF outbreaks and affected municipalities were detected using statistical methods for circular distributions. Spatial clustering of ASF-affected municipalities, as evaluated by join-count statistics, was significant in 1993 (Zjc = -3.0, P < 0.01) and 1994 (Zjc = -3.2, P < 0.01) but not in 1995 (Zjc = -0.6, P = 0.55) and 1996 (Zjc = -1.2, P = 0.23). Extensive pig farming and ASF were spatially co-distributed (kappa = 0.51, 95% CI = 0.33-0.70).

Synthesis and chromatographic properties of Se-carboxymethyl-selenohomocysteamine (2-carboxymethyl, 3-aminopropyl selenide).

Details are reported for the synthesis of Se-carboxymethylselenohomocysteamine from selenohomocysteamine and monochloroacetic acid. Data on its behaviour on paper and ion-exchange chromatography are also reported, which allow its identification.

Copper catalyzed alkaline autoxidation of selenocystamine.

In alkaline medium and in the presence of cupric ions selenocystamine undergoes autoxidation and is entirely transformed into selenohypotaurine. Among the different metal ions tested, Fe, Co, Ni, Cu, Ag, Mg, Mn, only cupric ions are effective in catalyzing the reaction. The reaction shows an optimum around pH 13. In most respects the autoxidation of selenocystamine is similar to the alkaline autoxidation of cystamine. Some data on the paper and ion exchange chromatographic behaviour of selenohypotaurine and selenotaurine are reported, as also details for the synthesis of selenotaurine.

Epidemiology of classical swine fever in Sardinia: a serological survey of wild boar and comparison with African swine fever.

A serological survey was carried out to establish the distribution of classical swine fever among wild boar in Sardinia, where that disease and African swine fever have been endemic in free-ranging domestic pigs and wild boar living in the mountainous areas of the province of Nuoro for several years. Blood samples were collected from 4752 wild boar shot during the period December 1988 to January 1992. An overall prevalence of 11 per cent was observed and the almost constant rate of about 9.8 per cent detected in the past three years indicates that the infection is well established. Wild boar seropositive to classical swine fever were found not only in the areas of the province of Nuoro where they share their habitat with free-ranging domestic pigs but also in other areas of the island where contacts between wild and domestic pigs are unlikely to occur. Therefore, transmission from wild boar to wild boar seems to play an important role in the spread and persistence of classical swine fever virus. In contrast, African swine fever virus is probably unable to persist in the wild boar population in the absence of the risk factor represented by their cohabitation with domestic free-ranging pigs infected with African swine fever.

[Chronic polyarthritis in a patient affected by sarcoidosis and chronic HCV infection. Case report and review of the literature]. / Poliartrite cronica in un caso di sarcoidosi ed epatopatia HCV correlata. Descrizione di un caso clinico e revisione della letteratura.

Sarcoidosis is a systemic granulomatous disease of unknown etiology that has a wide variety of clinical manifestation. Lung involvement may slowly undergo pulmonary fibrosis. Chronic sarcoid arthritis is a rare, usually non destructive arthropathy; may be a mono, oligo or polyarthritis. Knees, ankles, shoulders, wrists and small joint of the hands and feet may be involved. It can involve skin, eyes, exocrine glands such as salivary and lacrimal glands, and many other tissues. We describe the case of a 77 years old woman with a history of rhinopharyngitis with epistaxis and chronic laryngitis since youth; a dry mouth and throat, a erythematous, infiltrative skin lesion in the forehead and in the nape of the neck, a purple lesion of the left ear and nose, skin dystrophy of the hands from 30 years before. She underwent an operation for a left femoral fracture with hemotransfusion 14 years ago. Then she developed a polyarthritis of the small joints of the hands (II, III and IV right DIP, I, III, e V left DIP; III and V bilateral PIP), knees, tarsi, toes and left elbow. An HCV chronic hepatitis was discovered 6 years before. She is affected by productive cough, dysphonia, dyspnoea at rest, fever, headache and asthenia for over 5 years. Laboratory examination revealed leukopenia, HCV hepatitis with anti HCV, HCV-RNA, transaminases elevated and cryoglobulinemia. HCV may be involved in the etiopathogenesis of rheumatic diseases, lung fibrosis and may moreover contribute to the onset or progression of sarcoidosis; the possible pathogenesis is discussed.

[Neurotoxic syndrome of sheep due to the ingestion of plants of the Mediterranean scrub: clinical, histopathological, histochemical and ultrastructural observations]. / Sindrome neurotossica da ingestione di piante della 'macchia mediterranea' nell'ovino: osservazioni cliniche, istopatologiche, istochimiche ed ultrastrutturali.

A severe neurological disorder was observed during a very dry season, in sheep pasturing on land with mediterranean vegetation. During the crisis, lasting 5-20", ataxia and convulsions were the main clinical signs. Brain congestion, spongy appearance of the liver, interstitial nephritis and hyperpigmentation of visceral lymph nodes were the most important macroscopic lesions. Histopathological, histochemical and ultrastructural findings confirmed liver and kidney lesions and showed considerable neurovisceral storage of lipofuscin. Although further toxicological and experimental studies are needed to elucidate the origin of the disease, the Authors attribute the outbreak of the neurological syndrome to the ingestion of a neurotoxin containing plant.

Clinical presentation of thalassemia major due to homozygous beta (0)-thalassemia.

This study concerns the clinical presentation of all thalassemia major or intermedia in children admitted to out genetics department from January 1978 to December 1979. At electrophoresis and/or globin chain synthesis analysis on column chromatography. All these patients were shown to have homozygous beta(0)-thalassemia. The clinical severity was highly variable, dependent on age at presentation and when the patient first became dependent on transfusions. On the main reasons for this variability may be associated with thalassemia 1 or alpha-thalassemia 2 which have an incidence figure of 13 % in our population and hence also in these patients. Remarkable findings of the clinical and hematological picture were: (a) early spleen enlargement and growth deficiency; (b) MCV and MCH values intermediate between normals and beta-thalassemia heterozygotes; (c) normal mean hemoglobin A2 percentage and corpuscular concentration; (d) frequently increased transferrin saturation and ferritin levels; (e) presentation at age greater than 2 with hemoglobin levels about 9 g/dl usually associated with a milder course.

Chorionic villus sampling and acceptance rate of prenatal diagnosis.

In this paper, we compared the acceptance rate of fetal diagnosis for beta-thalassemia in three group of couples of Sardinian descent; the first counselled before DNA analysis was available, the second presenting after DNA analysis was introduced but too late for chorionic villus sampling and thus monitored by amniocyte DNA analysis and the third presenting within the first trimester after DNA analysis was introduced and thus in time for trophoblast DNA analysis. A higher proportion of couples from the latter group opted for fetal testing as compared to the 1st and 2nd group. These results indicate that in this population, introduction of 1st trimester diagnosis made prenatal testing acceptable to practically all counselled couples at risk.

Chromatographic separation of selenohypotaurine, selenotaurine, selenohomohypotaurine and selenohomotaurine.

By either paper or ion-exchange chromatography the two seleninic compounds selenohypotaurine and selenohomohypotaurine, and the two selenonic compounds selenotaurine and selenohomotaurine may be all separated from each other. On paper chromatography seleninic derivatives may be separated from the corresponding sulphinic compounds, while selenonic compounds show RF values similar to those of the corresponding sulphonic derivatives. These two latter types of compounds may be differentiated, however, since selenonic compounds liberate iodine from HI, while sulphonic compounds do not. Also by automated ion-exchange chromatography seleninic derivatives are well separated from the analogous sulphinic compounds, while selenonic compounds are eluted together with the corresponding sulphonic compounds.

A case of D13 ring chromosome.

A case of D ring chromosome identified with trypsin banding as a 13 with loss of the bands p12 and q34 is reported. The clinical features characteristically associated with the loss of these specific segments were present.

A case of extra small acrocentric bisatellited chromosome in a non mongoloid child.

A mentally retarded child with an extra small bisatellited acrocentric chromosome is described. The patient exhibited rather unspecific clinical signs such as strabismus, marked facial asymmetry, broad and prominent nasal bridge, hypertelorism, Brushfield's spots, malformed ears with atresia of the external auditory canal on the right side. Giemsa banding (R and G methods) did not allow a clear cytogenetic identification of the extra-chromosome. A tentative interpretation of the cytogenetic aberration as a trisomy of the proximal part of the long arm of chromosome 13 is discussed.

Control of homozygous beta-thalassemia by carrier screening and antenatal diagnosis in Sardinians.

This paper reports the present results of an ongoing program aimed at preventing homozygous beta-thalassemia by means of heterozygote screening and antenatal diagnosis in the Sardinian population. Screening based on the knowledge of carrier frequency and types of thalassemia prevalent in this population was designed to discover all heterozygotes except the few silent beta-thalassemia carriers. Most of the couples at risk were informed and accepted testing. Information was conveyed by mass media, midwives and marriage registry offices. Antenatal diagnosis was accepted by the majority of the couples counselled. The results of antenatal testing were very accurate. There was only one misdiagnosis out of 949 pregnancies tested. This risk of fetal loss was 7.5%. The program was highly effective, as shown by the decline of the incidence of the homozygous state from 1:205 live births in 1976 to 1:557 in 1981.

Auditory involvement in thalassemia major.

The auditory function of 75 children affected by homozygous beta0-thalassemia, managed with a low transfusion scheme and treated irregularly with low doses of desferrioxamine, and of 75 controls were examined. In 12 patients a mild bilateral conductive hearing impairment due to bony hypertrophy and/or adenoid hypertrophy was found. In 43 cases a moderate monolateral or bilateral sensory-neural hearing loss at high frequencies with recruitment phenomenon was observed. Ferritin levels were determined in a randomly chosen group of these patients with (14) and without heaing loss (11). In the subjects with sensory-neural hearing loss the mean ferritin levels were significantly higher than in those with no hearing defect. There was no obvious relation between sensory-neural damage on the one hand and Hb levels and unit of blood transfused on the other. The results of this study suggest that iron overload could be a cause of damage in the high frequency elements of the auditory mechanism. Intermittent hypoxia and slow 8th nerve compression due to bony hypertrophy as causes of auditory involvement are also discussed.