Abstract Ataxia-Teleangiectasia (A-T) is a neurodegenerative disorder due to mutations in ATM gene. ATM in the nucleus ensures DNA repair, while its role in the cytosol is still poorly clarified. ...Abnormal autophagy has been documented in other neurodegenerative disorders, thus we evaluated whether alteration in this process may be involved in the pathogenesis of A-T by analyzing the autophagic vesicles and the genes implicated in the different stages of autophagy. Through transmission electron microscopy (TEM) and immunofluorescence analysis we observed an accumulation of APs associated with a LC3 puncta pattern, and a reduced number of ALs. We also documented an increased expression of genes involved in AP and lysosome biogenesis and function, and a decrease of Vps18 expression, involved in their vesicular trafficking and fusion. mTORC1-controlled proteins were hyperphosphorylated in A-T, in keeping with an increased mTOR inhibitory influence of autophagy. Betamethasone is able to promote the degradation of SQSTM1, a biomarker of autophagy. Collectively, our results indicate that in cells from A-T patients, the APs maturation is active, while the fusion between APs and lysosomes is inappropriate, thus implying abnormalities in the cell-clearance process. We also documented a positive effect of Betamethasone on molecules implicated in autophagosome degradation.
Summary
Objective
Rate, reasons, and predictors of antiepileptic drug (AED) discontinuation were investigated in a well‐defined cohort of people with epilepsy to verify efficacy and tolerability of ...treatment up to 20 years from treatment initiation.
Methods
The history of AED usage in children and adults with epilepsy registered with 123 family physicians in an area of Northern Italy between 2000 and 2008 was recorded. Cumulative probabilities of AED withdrawal for specific reasons were estimated using cumulative incidence functions. The probabilities of withdrawing for terminal remission, and of achieving sustained remission while still on treatment, were also evaluated. The roles of sex, age at diagnosis, seizure types, duration at diagnosis, and syndrome were assessed with hazard ratios and 95% confidence intervals.
Results
Seven hundred thirty‐one of 747 individuals were treated with one or more AEDs during the disease course. The three commonest drugs were valproate, carbamazepine, and phenobarbital. Reported reasons for AED withdrawal were, in decreasing order, terminal remission, ineffectiveness, and adverse events. The probability of withdrawing the first AED for terminal remission was 1.0% at 1 year and increased to 20.0% at 20 years. Corresponding rates were 2.9% and 12.6% for ineffectiveness and 0.5% and 3.3% for adverse events. Reasons for withdrawal varied with individuals’ age, sex, disease characteristics, and drugs.
Significance
The initial AED given was retained in the majority of cases. Terminal remission, lack of efficacy, and adverse effects were, in decreasing order, the commonest reasons for AED discontinuation. Withdrawal could be predicted by age at diagnosis, sex, and clinical characteristics and varies among drugs.
Parkinson’s Disease (PD) is a neurodegenerative disorder characterized by the loss of dopaminergic neurons in the substantia nigra. Several studies were carried out in time on osteopathic treatments ...in patients with PD, demonstrating an improved motor function, albeit short lived. Our trial was aimed at assessing the potential efficacy of the Osteopathic Manipulative treatment (OMT) on mobility, posture, and gait in patients with moderate PD, as well as the importance of “taking charge” of the patient for the Quality of Life (QoL) of individuals with PD.
Material and methods. 32 subjects aged 76±7.284 were divided in two groups: group B (n=17) received the OMT, followed by the SHAM treatment; group A (n=15) received the SHAM treatment, then the OMt. the study was designed as a nine-week cross-over trial,
with group cross-overperformed after a one-week wash-out. All the subjects underwent a baseline neurological blind evaluation at four weeks and post cross-over. the ADL, IADL, GDS, and GpE scales were also administered.
Results. 16 subjects completed the study, 32 subjects participated in the first four weeks only. An improvement of the mean and standard deviation parameters was observed in the UpDRS and the tUG test in patients receiving the OMT. A decrease of the dysfunction degree was observed in both groups upon the osteopathic evaluation.
Conclusions. The data show the potential usefulness of the OMT in PD with respect to stiffness, mobility, posture and where the patient is taken charge of. Further studies will be needed to disprove the goal of the trial.
According to several laboratory protocols and specific conditions,
fertilization (IVF) dishes with culture media can be prepared 24
in advance compared to routine protocols. However, it is not clear ...if this procedure can affect embryological outcomes.
A nested case-control study was done in a cohort of couples undergoing IVF at the Infertility Unit of the ASST Lariana from August 2016 to July 2018. Cases were patients undergoing ovum pick up after a laboratory day off. Controls were patients undergoing ovum pick up after working days from Monday to Thursday. Culture media for oocyte culture and insemination were prepared about 42 and 18
before oocyte retrieval for cases and controls, respectively. Cases and controls were matched with a 1:2 ratio (for age, inseminated oocytes, length of stimulation). The "Good-Quality-Index" (GQI) was the main outcome to be compared between the two groups and was defined as good quality transferred or cryopreserved embryos on day 2 or 3+number of good quality blastocysts/inseminated oocytes.
A total of 76 cases and 152 matched controls were enrolled. The median GQI was equal to 33.0% (IQR: 20.0-50.0%) and 33.0% (IQR: 25.0-50.0%), in cases and controls, respectively (p=0.40). Study groups and GQI were not significantly correlated (correlation coefficient r=0.047, p=0.48). Main embryological parameters and cumulative pregnancy rates were similar between the two groups.
Our data support the vision that culture media can be prepared 24
in advance compared to routine protocols without affecting embryological outcomes.
Non-covalent (NP-1/3) and covalent (NP-A-1/3) pro-apoptotic SPION-Smac mimetic nano-conjugates antitumor agents are reported. The solution synthesis of key Smac mimetics, their support onto SPIONs ...through non-covalent adsorption (NP-1/3) or APTES-mediated covalent binding (NP-A-1/3), the analytical characterization of SPION-Smac mimetic conjugates, their target affinity in cell-free assays, and their cytotoxicity against tumor cells are thoroughly described.
Background and purpose
Seizures in most people with epilepsy remit but prognostic markers are poorly understood. There is also little information on the long‐term outcome of people who fail to ...achieve seizure control despite the use of two antiepileptic drugs (drug resistance).
Methods
People with a validated diagnosis of epilepsy in whom two antiepileptic drugs had failed were identified from primary care records. All were registered with one of 123 family physicians in an area of northern Italy. Remission (uninterrupted seizure freedom lasting 2 years or longer) and prognostic patterns (early remission, late remission, remission followed by relapse, no remission) were determined.
Results
In all, 747 individuals (381 men), aged 11 months to 94 years, were followed for 11 045.5 person‐years. 428 (59%) were seizure‐free. The probability of achieving 2‐year remission was 18% at treatment start, 34% at 2 years, 45% at 5, 52% at 10 and 67% at 20 years (terminal remission, 60%). Epilepsy syndrome and drug resistance were the only independent predictors of 2‐ and 5‐year remission. Early remission was seen in 101 people (19%), late remission in 175 (33%), remission followed by relapse in 85 (16%) and no remission in 166 (32%). Treatment response was the only variable associated with differing prognostic patterns.
Conclusion
The long‐term prognosis of epilepsy is favourable in most cases. Early seizure remission is not invariably followed by terminal remission and seizure outcome varies according to well‐defined patterns. Prolonged seizure remission and prognostic patterns can be predicted by broad syndromic categories and the failure of two antiepileptic drugs.
Abstract Drug-resistant epilepsy (DRE) is defined by the International League Against Epilepsy as a failure of adequate trials of two tolerated, appropriately chosen, and used antiepileptic drugs to ...achieve sustained seizure freedom. Our aim was to calculate the following: (1) the prevalence of active epilepsy and DRE in a well-defined population of Northern Italy and (2) the proportion of incident cases developing DRE. The study population (146,506; year 2008) resided in the province of Lecco, Northern Italy. The medical records of 123 general practitioners were reviewed to identify patients with epilepsy, diagnosed by a neurologist during the period 2000–2008. The point prevalence of active epilepsy and DRE was calculated on December 31, 2008. A total of 747 prevalent patients with epilepsy, 684 patients with active epilepsy, and 342 incident cases were identified. The frequency of DRE was 15.6% (107/684) of all active epilepsies and 10.5% (36/342) of incident cases. The point prevalence was 0.73 per 1000. The standardized prevalence of DRE was 0.7 per 1000 (Italian population) and 0.8 per 1000 (world population). Our data indicate that 1/6 patients with active epilepsy in the general population has DRE, and 1/10 patients with newly diagnosed epilepsy will develop DRE within nine years from the diagnosis.
The allergenicity of seed storage proteins, the major components of edible legume seeds, may cause serious reactions in both children and adult population. Updated methodologies for evaluation of the ...activity of these proteins are needed. In this paper we used two-dimensional (2D) electrophoretic techniques to investigate the immuno-cross-reactivities of anti Ara h 3 basic subunit IgG to the seed proteomes of three legume species, namely, peanut, soybean, and lupin. The seed proteins, extracted with two different procedures, were separated by 2D electrophoresis, and the electrophoretic maps were analyzed by Western blot. In peanut proteome the antibodies strongly reacted with the 23 kDa polypeptides, corresponding to Ara h 3 basic isoforms, the antigen they were raised to, and three unidentified acidic polypeptides near 45 kDa. Remarkable cross-reactivities with lupin and soybean Ara h 3 homologous polypeptides and nonrelated proteins, namely, lupin conglutin γ and soybean Bg7S, were detected. Therefore, these proteins may be regarded as new putative allergens. The present findings show the potentiality of 2D electrophoresis in the identification of food allergens and open the way to the traceability of the new cross-reacting proteins in the food chain. Keywords: Arachis hypogeae; Glycine max; Lupinus albus; legume seeds; peanut allergens; IgG; IEF/SDS−PAGE; cross-reactivity
Activating mutations in v‐Ha‐ras Harvey rat sarcoma viral oncogene homolog (HRAS) have recently been identified as the molecular cause underlying Costello syndrome (CS). To further investigate the ...phenotypic spectrum associated with germline HRAS mutations and characterize their molecular diversity, subjects with a diagnosis of CS (N = 9), Noonan syndrome (NS; N = 36), cardiofaciocutaneous syndrome (CFCS; N = 4), or with a phenotype suggestive of these conditions but without a definitive diagnosis (N = 12) were screened for the entire coding sequence of the gene. A de novo heterozygous HRAS change was detected in all the subjects diagnosed with CS, while no lesion was observed with any of the other phenotypes. While eight cases shared the recurrent c.34G>A change, a novel c.436G>A transition was observed in one individual. The latter affected residue, p.Ala146, which contributes to guanosine triphosphate (GTP)/guanosine diphosphate (GDP) binding, defining a novel class of activating HRAS lesions that perturb development. Clinical characterization indicated that p.Gly12Ser was associated with a homogeneous phenotype. By analyzing the genomic region flanking the HRAS mutations, we traced the parental origin of lesions in nine informative families and demonstrated that de novo mutations were inherited from the father in all cases. We noted an advanced age at conception in unaffected fathers transmitting the mutation. Hum Mutat 28(3), 265–272, 2007. Published 2006 Wiley‐Liss, Inc.
Several proteins have been identified as peanut allergens; among them, Ara h 1 (7S globulin) and Ara h 2 (2S globulin) are usually considered the major allergens.
To identify the major allergens in a ...group of children selected for their specific pattern of immunoreactivity.
We identified the dominant allergen by using (1) amino acid sequencing of the bands that show the strongest IgE immunoreactivity in 1-dimensional electrophoresis and immunoblotting and (2) specific animal IgGs raised against the dominant immunoreactive band to pinpoint the allergen(s) in peanut proteins separated by 2-dimensional electrophoresis and immunoblotting. To confirm these data, we further examined the peanut proteome using serum samples from the children with the unusual immunoreactivity.
We found a group of children with marked peanut allergy who are specifically sensitized to the basic subunit of Ara h 3 (11S globulin family).
That the dominant immunoreactivity in these patients is in a basic subunit of Ara h 3 was unexpected, because previous studies had indicated that Ara h 3 was only a minor peanut allergen and that the identified allergenic epitopes occurred mainly in the acidic Ara h 3 subunit.
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