A peculiar family with recurrent self-limited epileptic syndrome and associated developmental disorders in six girls.

We describe a complex family with two couples (two sisters who married two brothers) with consistent social and neuropsychiatric problems, originally from Sardinia. Each couple had three daughters, which shared electroclinical epileptic syndrome and developmental disorders. All patients suffered from mild to moderate intellectual disability, speech difficulties and behavioural disorders. Four out of six patients had epilepsy onset between 3 and 4 years of age. The epileptic history almost reflected the typical clinical course of a self-Limited Focal Epilepsy of Childhood. However, our patients don't have the complete features characteristic of one of the four specific self-Limited Focal Epilepsies of Childhood; a progressive evolution into a Developmental and/or Epileptic Encephalopathy with spike-wave activation in sleep was observed in the two older sister of the first family, which developed more severe developmental disorder too. In the other epileptic patients, improvement of EEG pattern was not coincident with an improvement of the developmental disorders. Brain MRI, performed in three patients, showed normal findings. Genetic analysis carried out so far (SNP-array, study of Runs of homozygosity, FMR1 triplet-repeat primer-PCR assay, Next Generation Sequencing based gene panel for epilepsy and neurodevelopmental disorders and Exome Sequencing), did not provide useful elements for an aetiological diagnosis.

Barotraumatic orbital emphysema of rhinogenic origin in a breath-hold diver: a case report.

Orbital emphysema is a well-recognized complication of fractures involving the orbit. Commonly, it occurs when high pressure develops in nasal cavity as during nose blowing, coughing or Valsalva's maneuver and usually occurs in the subcutaneous tissues. We report the case of a young breath-hold diver who developed spontaneous, non compressive orbital emphysema during underwater fishing, with a maximal depth of 25-30 meters in the Sardinian sea. He was otherwise healthy, without previous cranio-facial trauma and nasosinusal diseases or surgery were not present in the history. When he was referred to our attention the patient presented right eyelid ptosis but diplopia and vision impairment were absent. Computer tomography scans showed subcutaneous air in the right upper eyelid and around the eyeball, particularly near the orbit's roof but optic nerve area, intraconal, was free of air. A dehiscence in lamina papyracea was evident. In our opinion, this has been the point of air entry into the orbit. A supportive therapy was advised and two weeks later the emphysema was recovered completely and the subject was symptoms free. The literature has been revised and to our knowledge no previous cases of barotraumatic orbital emphysema, in a breath-hold diver, are referred.

Thalassaemia and glucose-6-phosphate dehydrogenase screening in 13- to 14-year-old students of the Sardinian population: preliminary findings.

OBJECTIVES: In this paper we describe the outline and results of a 7-year screening programme for thalassaemias and glucose-6-phosphate dehydrogenase (G6PD) deficiency in 13- to 14-year-old students from the Sardinian population. METHOD: This programme had several steps: formal education on thalassaemia, request of informed consent by parents, blood testing and genetic counselling. RESULTS: Out of 63,285 subjects tested, 6,521 (10.3%) were heterozygotes for beta-thalassaemia, 16,175 (25.6%) for alpha-thalassaemia and 101 were carriers of a haemoglobin variant. One thousand four hundred and twenty (16.4%) males were hemizygotes for G6PD deficiency and 1,893 (20.6%) females were heterozygotes. CONCLUSION: The uptake of the programme was remarkably high and homogeneous across the island, indicating and confirming a great interest of the Sardinian population in any initiative directed at the prevention of homozygous beta-thalassaemia.

Laser-based diagnostics applied to the study of BN nanotubes synthesis.

The boron nitride nanotubes (BNNTs) synthesis, using CO2-laser vaporization of a BN target under nitrogen gas, is investigated by UV-laser induced fluorescence (LIF) of the vapor phase and UV-Rayleigh scattering (RS) of the gas-suspended nanoparticles. The LIF signal from B atoms is mainly detected in the 1.5 mm-thick region above the BN target. It originates from a boron-rich vapor region confined near the hot boron droplet formed at the target surface. Then, recombination between hot boron and N2 gas occurs through a fast condensation process as revealed by both the depletion of B atoms from the vapor phase and the RS signal arising from the grown BN nanoparticles. Fluorescence spectra exhibit a strong peak at 250 nm due to boron fluorescence and mainly to nanoparticles Rayleigh scattering. A narrow peak is observed at 210 nm and a broader peak at 189 nm. These bands are tentatively assigned to fluorescence or photoluminescence (PL) from gaseous or solid BN species respectively since both gas and solid phases coexist in the plume due to the rapid cooling process. Two very weak bands occur at 308 nm and 350 nm. They are related to PL of defects bands from BN nanostructures on the basis of ex situ PL spectra of h-BN crystallites and multi-wall BNNTs. Detection of oxygen impurities is shown feasible through LIF from BO radical which is detected just above the BN target evaporated under vacuum pressure (approximately 1 mbar). An optical diagnostic strategy is demonstrated from these first in situ observations during BNNTs synthesis.

Spatial evolutions of Co and Ni atoms during single-walled carbon nanotubes formation: measurements and modeling.

Spatial investigations of nickel and cobalt atoms and of C2 and C3 radicals are performed by laser induced fluorescence (LIF) in a continuous CO2 laser-vaporization reactor during the synthesis of single-walled carbon nanotubes. The chemical composition of the gas vaporized from bimetallic Ni/Co catalysts-carbon targets is determined using a chemical kinetic model. In this model, the evolution of Ni and Co atoms is driven by kinetics of condensation/evaporation process of pure metal clusters. Metal-carbon clusters are assumed to form from soot particles (C80) and 128-atom metal clusters. Spatial profiles of Ni and Co atoms obtained by LIF are compared with the calculations to validate the modeling and to adjust the input data. The value of the initial molar fraction of carbon-metal mixture diluted in helium is determined through a parametric study. Good agreement is found between the measured and the calculated evolution of Ni for a molar fraction of the helium diluent ranging from 10 to 15%. To fit the spatial profile of Co, the activation energy is adjusted in the evaporation rate, changing the cobalt dimer bond energy. The latter is found to be largely uncertain; and three values are tested: 167, 208, and 230 kJ x mol(-1). From comparison, the activation energy is found to be 208 kJ x mol(-1). However, the C2 LIF profiles show that the depletion of C2 is accelerated when cobalt is present. The observed Co evolutions suggest that small Co-C clusters are easier and/or faster to form compared to Ni-C clusters.

Optical transitions in single-wall boron nitride nanotubes.

Optical transitions in single-wall boron nitride nanotubes are investigated by means of optical absorption spectroscopy. Three absorption lines are observed. Two of them (at 4.45 and 5.5 eV) result from the quantification involved by the rolling up of the hexagonal boron nitride (h-BN) sheet. The nature of these lines is discussed, and two interpretations are proposed. A comparison with single-wall carbon nanotubes leads one to interpret these lines as transitions between pairs of van Hove singularities in the one-dimensional density of states of boron nitride single-wall nanotubes. But the confinement energy due to the rolling up of the h-BN sheet cannot explain a gap width of the boron nitride nanotubes below the h-BN gap. The low energy line is then attributed to the existence of a Frenkel exciton with a binding energy in the 1 eV range.

Cytomorphologic and ultrastructural study of nasal mucosa by means of brushing: a comparison between asymptomatic and rhinitic subjects.

The authors compared the cytomorphologic and ultrastructural features of nasal epithelium collected by means of brushing from asymptomatic subjects with these of patients affected by nasal polyposis (NP), allergic (AR) and non-allergic rhinitis (NAR). A brushing of nasal epithelium was taken from each member of both groups, and analysed under light and electron microscopy. The observation showed normal ciliary patterns and preserved intercellular joints in the great majority of asymptomatic subjects, while in all subjects of the pathologic group the junctions appeared variously damaged or absents, with ciliary abnormalities. The damage to the intercellular joints, rather than the alterations of ciliary patterns, seemed to represent in this study the border between the absence of symptoms and the presence of chronic inflammation. Therefore, a reduced tightness of the intercellular joints could contribute to the impairment of the mucociliary clearance, priming the vicious circle that leads to the condition of chronic inflammation.

In-situ optical analysis of the gas phase during the formation of carbon nanotubes.

A reactor has been developed at ONERA to investigate the gas phase during carbon nanotube formation by laser-induced fluorescence (LIF), Laser-induced incandescence (LII), coherent anti-Stokes Raman Scattering (CARS), and emission spectroscopy. Continuous vaporization is achieved with a continuous wave CO2 laser. Optimized conditions are used for single-walled nanotube growth, that is, a graphite target doped with 2 atom % Ni and 2 atom % Co, helium as buffer gas at a flow rate of 50 ml/s, and a pressure of 300 hPa. Temperature profiles are measured by CARS on H2, and soot images are obtained by LII in the hot carbonaceous flow. LIF and spontaneous emission of the C2 radical and Ni and Co atoms are presented. Spectral investigations are conducted at 3100 and 3200 K to have an optimized pair of excitation/detection wavelengths. Spatial investigations of the relative concentrations in the hot carbonaceous flow are performed up to 3500 K. The concentrations are measured as a function of target temperature. Two regimes of vaporization are observed. Vaporization is slow up to 3350 K and becomes much faster above this temperature. The fast regime in the 3350-3500 K range corresponds to the observed spatial extent of the metal vapors region. At 3500 K, the C2 profiles obtained with and without catalysts are very different as a result of carbon coalescence as well as carbon dissolution into the metal nanoparticles when these are present in the gas phase. The shape of the C2 profile can be related to nanotube formation and growth at a target temperature of 3500 K.

Pulmonary stenosis and atresia with intact ventricular septum during prenatal life.

AIM: To identify fetal echocardiographic characteristics predictive of perinatal outcome in cases with a prenatal diagnosis of pulmonary stenosis or pulmonary atresia. PATIENTS AND METHODS: We retrospectively reviewed the records and the videotapes of all the cases of pulmonary stenosis and pulmonary atresia diagnosed at our institutions between 1990 and 1999. The following measurements were obtained: diameters of right and left atria and ventricles and ventricular wall thickness; main pulmonary artery and aortic root diameter; direction of flow through the atrioventricular, aortic and pulmonary valves and through the ductus arteriosus. Perinatal outcome and follow-up of the survivors were available in each case. RESULTS: There were 21 cases of pulmonary atresia. Eleven were diagnosed before 24 weeks and nine of them (82%) underwent termination of pregnancy. The survival rate was 50% among the 12 fetuses born at term. None of the fetuses that survived had a large right ventricle, while this was a finding in 50% of those that died. Among the fetuses that died, 83% had a hypertrophic right ventricular wall compared to 33% of the survivors. There were 12 cases of pulmonary stenosis. Three cases were diagnosed before 24 weeks but none underwent termination of pregnancy. All the fetuses with pulmonary stenosis were born at term and four died in the perinatal period. The survival rate was thus 66.6% (8/12). Three (75%) of the fetuses that died had reversed flow in the ductus arteriosus compared with one of the fetuses that survived. CONCLUSION: Our data suggest that a grossly enlarged right ventricle and/or a hypertrophied right ventricular wall in cases of pulmonary atresia and reversed flow in the arterial duct in cases of pulmonary stenosis are likely indicators of a poor prognosis.

Ultrasound evaluation of uterine wound healing following laparoscopic myomectomy: preliminary results.

The purpose of our work was to study the evolution of the uterine scar following laparoscopic myomectomy, as imaged by ultrasonography and Doppler velocimetry of the uterine arteries. We prospectively studied 30 patients. In the first phase, 15 patients were submitted to two-dimensional (2D) endovaginal ultrasound on day -1, 1, 7, 30 and 60 (surgery = day 0). In the second phase an additional 15 patients were studied by both 2D ultrasound and by Doppler velocimetry. The resistance index (RI) was calculated from the flow velocity waveform of the uterine arteries, at the origin of their ascending branch. Only one ultrasonic pattern was found, which was a dense echogenic area having an ill-defined, heterogeneous texture. In one case a small anechoic area (1 cm) was detected in the scar, possibly due to a haematoma. The evolution of uterine healing showed a progressive reduction in the size of the scar. On day 1 its mean diameter was 37.04% less than the myoma diameter and on day 30 71.7% less. The difference was significant at P < 0.001. A further significant (P < 0.001) reduction was found at day 60 in the 15 patients studied in phase I. On both day 1 and day 30 following surgery, there was no correlation between the sizes of the myoma and the scar. There was a statistically significant increase (P < 0.01) in the RI value of the ipsilateral uterine artery from 0.64 on day -1 to 0.79 on day 1. On day 30, 12/15 (80%) cases had RI values ranging between 0.80 and 0.98, while in three cases there was absence of end diastolic flow. The RI values of the contralateral uterine artery were high (0.90) before surgery and did not change afterwards. There was no correlation between the size of the myoma and the increase in the uterine artery RI value following surgery. Considering the velocimetric findings, 30 days are a reference point for assessing the healing process. Ultrasound imaging and Doppler velocimetry can be used for studying the evolution of the uterine scar following myomectomy.

Two novel mutations (10410 T-->G; 10296 del C) at carboxy-terminus of the dystrophin gene associated with mental retardation. Mutations in brief no. 149. Online.

In this study we have carried out a mutational screening of exons 62-79 of the dystrophin gene by SSCP in 38 Italian patients with DMD/BMD and found two novel mutations at exon 70, in 2 mentally retarded DMD patients.

Elevation of serum creatine kinase as the only manifestation of an intragenic deletion of the dystrophin gene in three unrelated families.

This study reports three children from three unrelated families, aged from 9 to 12 years, who were investigated because of the incidental finding of elevated serum creatine kinase (CK) levels and were found to have a dystrophinopathy. The molecular defect consisted of a deletion of variable extent within the central rod domain of the dystrophin gene, involving either exons 32-44 or 48-51 or 48-53. In each family we found the same deletion in at least one adult male relative aged from 40 to 77 years, who was either completely asymptomatic or had very mild muscle involvement (thin muscles and/or mild scoliosis), with normal or borderline CK levels. This study suggests once again that deletions of the central rod domain of dystrophin may be associated with elevation of serum CK as the only manifestation and that prediction of the clinical severity based solely on the molecular findings should be interpreted with caution.

Dystrophin gene abnormalities in two patients with idiopathic dilated cardiomyopathy.

Two new cases of dilated cardiomyopathy (DC) caused by dystrophinopathy are reported. One patient, a 24 year old man, had a family history of X linked DC, while the other, a 52 year old man, had sporadic disease. Each had abnormal dystrophin immunostaining in muscle or cardiac biopsy specimens, but neither had muscle weakness. Serum creatine kinase activity was raised only in the patient with familial disease. Analysis of dystrophin gene mutations showed a deletion of exons 48-49 in the patient with familial DC and of exons 49-51 in the other. Dystrophin transcription in cardiac tissue from the patient with sporadic disease showed abundant expression, predominantly of the muscle isoform. This study, together with previous reports, suggests that some patients with DC have a dystrophinopathy that can be diagnosed using a combination of biochemical and genetic analyses.

A report on 528 intragenic deletions detected in DMD and BMD patients by an Italian collaborative study.

The results of a collaborative study involving about one third of the total DMD and BMD cases living in the Italian territory are reported. The analysis of the breakpoint frequency by intron revealed significant differences among regional groups of DMD patients (for introns 2, 11 and 50 in Sardinia and for introns 9 and 45 in northeastern Italy), whereas no regional differences were observed among regional groups of BMD patients. These differences involve the same Italian regions which previous studies, performed by different markers, identified as "genetically differentiated". The data support the possibility of a differential distribution among populations of some intronic sequences, facilitating the origin of deletion breakpoints within the dystrophin gene.

Identification of a novel T-insertion polymorphism at the DMD locus.

We have identified a novel T-insertion polymorphism located in the second intron of the dystrophin gene. This polymorphism should prove useful in linkage studies in Duchenne and Becker muscular dystrophy families in addition to the previously described markers.

Germinal mosaicism in a Duchenne muscular dystrophy family: implications for genetic counselling.

In this study we describe a three-generation family in which two siblings were affected by Duchenne muscular dystrophy (DMD). Immunohistochemical analysis of muscle dystrophin and haplotype analysis of the DMD locus revealed that the X chromosome carrying the DMD gene was transmitted from the healthy maternal grandfather to his three daughters, including the proband's mother. These findings indicate that the grandfather was a germinal mosaic for the DMD gene. The definition of the carrier status in two possible carriers led us to give accurate genetic counselling and to prevent the birth of an affected boy. The results of this study demonstrate the usefulness of haplotype analysis and immunohistochemical muscle dystrophin studies to detect hidden germinal mosaicism and to improve genetic counselling.

Diagnosis of DMD carrier status in a family with no known affected males.

A 30-year-old woman and her two-year-old daughter were found by chance to have moderately raised serum creatine kinase (CK) levels. Since the mother was pregnant, the authors investigated the possibility that the two females were carriers of the common Duchenne muscular dystrophy (DMD) gene. No immunohistochemical abnormality was detected in the mother, but in the daughter a clear mosaic pattern of dystrophin positive and negative fibres was found, indicating carrier status for DMD. These data indicate that a diagnosis of DMD carrier status can be made even in families without a positive history for this disorder; therefore, immunocytochemical studies, using antidystrophin antibodies, should be performed on all females with raised CK levels, including the youngest.