Response of cellular antioxidants to ozone in cultured human fibroblasts

There are some evidence that oxygen-ozone therapy may be beneficial in several human diseases. However the biochemical and pharmacodynamic mechanisms have not been elucidated yet. The ozone therapy is closely related to the "oxidative stress" conception because it generates an effect through the stimulation of antioxidant defence systems against the activity of the noxious oxidative species. Maybe ozone can exert its protective effect by means of an oxidative preconditioning, stimulating and/or preserving the endogenous antioxidant systems. The aim of this work is to evaluate the effects of ozone exposure on antioxidant cellular system in culture of human fibroblast. Normal primary fibroblasts were obtained from healthy donor. Oxygen (O2) or mixture of ozone (O2-O3) were administered (40 or 80 µg/ml) directly on medium for 180 seconds. The fibroblast are tripsinized 30 minutes, 3 hours, 3 day after gaseous exposure. The levels of glutathione reduced (GSH) and oxidized (GSSG), ascorbic acid (AA) and malondialdehyde (MDA) were measured in cells, moreover the concentrations of nitrixc oxide (NO) were evaluated in the medium. The exposures to O2-O3 comparison to control group (cells no exposed) produce an significant increase of GSH (+150 % at 30min, P<0.001), GSSG (+62% at 30min P<0.05), NO (+100% at 3h, P<0.05) levels and a diminution of AA concentrations (-84% P<0.001 after 30 min) and no production of MDA. Our results show that ozone doesn’t induce stress oxidative at used doses and times. On the basis of preliminary data we postulated that ozone administration would promote an oxidative preconditioning preventing the cellular damage. We speculate that effectiveness of ozone could be related to its action on endogenous antioxidants and pro-oxidants balance in favour of antioxidants, explaining thus the good clinical results obtained with ozone therapy

Role of novel and rare nucleotide substitutions of the β-globin gene

The Laboratory for Molecular Prenatal Diagnosis of Hemoglobinopathies at the Villa Sofia-Cervello Hospital in Palermo, Italy, carries out an intensive screening program aimed at identifying the healthy carriers of thalassemia and, consequently, the couples at risk of bearing an affected fetus. The diagnostic process is basically divided into two phases: i) hematologic and hemoglobin data; ii) molecular analysis of globin genes and, when possible, a genetic study of the family. Since 2003, we have been performing DNA sequence analysis on those cases in which classical molecular methods failed to give a complete diagnostic response, particularly in phenotypes with borderline values of HbA2 with mild or absent microcytosis. During ten years of screening activities (from 2003 to 2012), twenty- seven unknown or rare nucleotide changes of the &beta;-globin gene have been identified; hematologic and hemoglobin data have been carefully evaluated and, wherever possible, we have conducted a family study to evaluate whether a phenotypic expression could be associated to these nucleotide changes. Because of the limited numbers of cases for each mutation, the significance of these nucleotide substitutions has still not been fully clarified, and this raises a number of questions that need to be answered when carrying out appropriate genetic counseling for couples presumed to be at risk.&nbsp;意大利巴勒莫Villa Sofia-Cervello医院血红蛋白病分子产前诊断实验室进行密集的筛选程序，旨在识别健康的地中海贫血携带者和有怀上地中海贫血胎儿风险的夫妇。 诊断过程基本上分为两个阶段：1）血液及血红蛋白数据；2）珠蛋白基因分子分析以及家族遗传研究（如有可能）。 自2003年以来，我们已对这类病例进行DNA序列分析：传统的分子方法无法给出完整的诊断响应，尤其是有轻微小红细胞症或缺乏小红细胞症的HbA2临界值表型。 通过十年筛选活动（2003年至2012年），已识别出&beta;珠蛋白基因的27个未知或罕见的核苷酸变化；仔细评估过血液和血红蛋白数据，在可能的情况下，我们已进行家系研究，以确定表型表达是否与这些核苷酸变化相关。由于只有有限的突变案例，所以至今尚未完全证实这些核苷酸置换的重要性，并对假定存在患病风险的夫妇开展适当的基因遗传咨询时，因此引起了大量需要解答的问题。</p

Cutis verticis gyrata and Noonan syndrome: report of two cases with pathogenetic variant in SOS1 gene

Background Noonan and Noonan-like syndromes are multisystem genetic disorders, mainly with autosomal dominant trasmission, caused by mutations in several genes. Missense pathogenetic variants of SOS1 gene are the second most common cause of Noonan syndrome (NS) and account approximately for 13% to 17% of cases. Subjects carrying a pathogenetic variant in SOS1 gene tend to exhibit a distinctive phenotype that is characterized by ectodermal abnormalities. Cutis verticis gyrata (CVG) is a rare disease, congenital or acquired, characterized by the redundancy of skin on scalp, forming thick skin folds and grooves of similar aspect to cerebral cortex gyri. Several references in the literature have reported association between nonessential primary form of CVG and NS. Case presentation we report two cases of newborns with CVG and phenotype suggestive for NS who have been diagnosed to harbour the same pathogenetic variant in SOS1 gene. Conclusions previously described patients with NS presenting CVG had received only clinical diagnosis. Therefore we report the first patients with CVG in which the clinical suspicion of NS is confirmed by molecolar analysis

Prenatal diagnosis of hemoglobinopathies: from fetoscopy to coelocentesis

Prenatal diagnosis of hemoglobinopathies involves the study of fetal material from blood, amniocytes, trophoblast coelomatic cells and fetal DNA in maternal circulation. Its first application dates back to the 70s and it involves globin chain synthesis analysis on fetal blood. In the 1980s molecular analysis was introduced as well as amniocentesis and chorionic villi sampling under high-resolution ultrasound imaging. The application of direct sequencing and polymerase chain reactionbased methodologies improved the DNA analysis procedures and reduced the sampling age for invasive prenatal diagnosis from 18 to 16- 11 weeks allowing fetal genotyping within the first trimester of pregnancy. In the last years, fetal material obtained at 7-8 weeks of gestation by coelocentesis and isolation of fetal cells has provided new platforms on which to develop diagnostic capabilities while non-invasive technologies using fetal DNA in maternal circulation are starting to develop

Optogenetic Stimulation of Prelimbic Pyramidal Neurons Maintains Fear Memories and Modulates Amygdala Pyramidal Neuron Transcriptome

Fear extinction requires coordinated neural activity within the amygdala and medial prefrontal cortex (mPFC). Any behavior has a transcriptomic signature that is modified by environmental experiences, and specific genes are involved in functional plasticity and synaptic wiring during fear extinction. Here, we investigated the effects of optogenetic manipulations of prelimbic (PrL) pyramidal neurons and amygdala gene expression to analyze the specific transcriptional pathways associated to adaptive and maladaptive fear extinction. To this aim, transgenic mice were (or not) fear-conditioned and during the extinction phase they received optogenetic (or sham) stimulations over photo-activable PrL pyramidal neurons. At the end of behavioral testing, electrophysiological (neural cellular excitability and Excitatory Post-Synaptic Currents) and morphological (spinogenesis) correlates were evaluated in the PrL pyramidal neurons. Furthermore, transcriptomic cell-specific RNA-analyses (differential gene expression profiling and functional enrichment analyses) were performed in amygdala pyramidal neurons. Our results show that the optogenetic activation of PrL pyramidal neurons in fear-conditioned mice induces fear extinction deficits, reflected in an increase of cellular excitability, excitatory neurotransmission, and spinogenesis of PrL pyramidal neurons, and associated to strong modifications of the transcriptome of amygdala pyramidal neurons. Understanding the electrophysiological, morphological, and transcriptomic architecture of fear extinction may facilitate the comprehension of fear-related disorders