Free and glycosylated green leaf volatiles, lipoxygenase and alcohol dehydrogenase in defoliated Nebbiolo grapes during postharvest dehydration

none7noBackground and Aims: Nebbiolo grapes are used to produce Sfursat wine, following partial dehydration. This research aimed to clarify the influence of fruit exposure to light and postharvest water loss on the concentration of green leaf volatiles (GLVs) and lipoxygenase (LOX) and alcohol dehydrogenase (ADH) activity of grapes. Methods and Results: Nebbiolo grapes from Control vines (no defoliation) (ND) and from vines defoliated at fruitset (DFS) or defoliated post-veraison (DPV) were harvested at about 23°Brix and dehydrated at 10 and 20°C, 60% RH and air flow of 1.5 m/s. Berries were sampled at 10 and 20% mass loss (ML). Significant differences in crop yield, bunch mass and berry mass were observed. As expected, the higher the dehydration temperature, the faster the dehydration process: 20% ML at 20°C occurred between 18 and 25 days, the shortest time corresponding to ND and the longest to DFS; at 10°C, the dehydration lasted between 27 and 32 days. At 10°C, the ADH activity was almost double that at 20°C, and in DFS was much higher than in other samples. At harvest, LOX did not show any difference among the samples, while at 10°C and 10% ML, the enzyme activity increased significantly and then declined at 20% ML, especially in defoliated samples. At harvest, the total free GLVs associated with the metabolism of lipid oxidation were 9434, 7212 and 11 656 μg/kg dry weight (DW) in ND, DFS and DPV samples, respectively; the total bound GLVs lipid-derived were 7599, 18 486 and 15 409 μg/kg DW in ND, DFS and DPV samples, respectively. During dehydration at 10°C, the ML induced ADH + LOX activity, especially in defoliated samples, but the bound GLVs, produced by defoliation, greatly decreased. Conclusions: Defoliation affected the response of Nebbiolo grapes to dehydration temperature: postharvest cold stress (10°C) and ML induced glycosylation of GLVs, alcohol formation (via ADH) and membrane oxidation (via LOX); a further stress effect was observed with leaf removal, regardless of the time of application. Significance of the Study: The timing of defoliation and postharvest dehydration temperature are significant factors to mitigate the postharvest stress response of Nebbiolo grapes.openPiombino P.; Genovese A.; Rustioni L.; Moio L.; Failla O.; Bellincontro A.; Mencarelli F.Piombino, P.; Genovese, A.; Rustioni, L.; Moio, L.; Failla, O.; Bellincontro, A.; Mencarelli, F

Porencephaly in an Italian neonate with foetal alcohol spectrum disorder: A case report

INTRODUCTION: Foetal alcohol spectrum disorder (FASD) is a complex malformative disease caused by the teratogenic effect of alcohol consumed during pregnancy. Mothers are frequently reluctant to admit alcohol consumption during pregnancy. During infancy and particularly during neonatal period, differential diagnosis is difficult. PATIENT CONCERNS: This case is represented by an Italian neonate boy small for gestational age, born by caesarean section at a gestational age of 37 weeks + 6 days by neglect and single-parent pregnancy. On physical examination, he presented particular facial features: microcephaly, epicanthal folds, flat midface, low nasal bridge, indistinct philtrum, and thin upper lip; moreover, examination revealed a macro-penis and recurvation without evidence of glans. DIAGNOSIS: Echocardiogram showed an inter-ventricular defect of medium-muscular type and brain magnetic resonance imaging showed asymmetry of the cerebral hemispheres with hypoplasia of the left cerebral hemisphere, dilatation of the left ventricle, cerebrospinal fluid cavity, and porencephaly. INTERVENTIONS: We investigated the ethylglucuronide (EtG) concentration in the neonate's hair by liquid chromatography-tandem mass spectrometry and we detected EtG in the infant's hair (normal value, 30 pg/mg), demonstrating prenatal alcohol exposure. OUTCOMES: In this neonate, EtG measure in hairs permitted the diagnosis of FASD, so allowing to exclude genetic diseases associated with similar clinical findings. After this result the mother admitted that she drunk alcohol during pregnancy (she declared 3 glasses of wine every day). At the age of 6 months, the child showed a moderate neurodevelopmental delay. CONCLUSION: This case shows that FAD should be considered in neonates with rare neurological diseases as porencephaly. In neonates and infants born to a mother who did not report alcohol use, EtG measure in hairs can significantly improve diagnosis of FASD, so allowing to exclude genetic diseases associated with similar clinical findings

Postharvest Ozone Fumigation of Grapes (cv Sangiovese) Differently Affects Volatile Organic Compounds and Polyphenol Profiles of Berries and Wine

Consumers are more and more oriented towards the purchase of safer food and beverages, which is pushing the wine sector to find alternatives to the use of sulfur dioxide. Ozone (O3) is already applied in the wine industry to produce sulfur dioxide-free wines through the patented method Purovino®. The aim of this two-year study was that of evaluating whether the postharvest treatment of grapes with ozone affects volatile organic compounds (VOCs) and polyphenol profile in berries, and in turn, wine composition. Grape bunches (Vitis vinifera L.) of cv Sangiovese were fumigated overnight with gaseous ozone (max 20 g·h−1 with 6% w.w−1 of ozone) in a cold room at 4°C (±0.5). After treatment, grapes were processed into wine. In grapes, ozone treatments increased total polyphenol and flavonoid content and upregulated specific genes (phenylalanine ammonia lyase, VvPAL, flavanol synthase 1, and VvFLS1) involved in polyphenol biosynthesis. Wine obtained from ozone-treated grapes had higher flavanol content than the control. Fumigation only slightly affected the different VOC classes of grapes and wine, including aroma compounds derived from the lipoxygenase (LOX) pathway. Although a season-dependent effect was observed, results showed that postharvest ozone treatments applied to avoid the use of sulfur dioxide introduced limited but, in general, positive modifications to grape and wine composition. This information provides assurance to winemakers that the maintenance of wine quality and typicity will be guaranteed when using ozone treatments

Juvenile moyamoya and craniosynostosis in a child with deletion 1p32p31: Expanding the clinical spectrum of 1p32p31 deletion syndrome and a review of the literature

Moyamoya angiopathy (MA) is a rare cerebrovascular disorder characterised by the progressive occlusion of the internal carotid artery. Its aetiology is uncertain, but a genetic background seems likely, given the high MA familial rate. To investigate the aetiology of craniosynostosis and juvenile moyamoya in a 14-year-old male patient, we performed an array-comparative genomic hybridisation revealing a de novo interstitial deletion of 8.5 Mb in chromosome region 1p32p31. The deletion involved 34 protein coding genes, including NF1A, whose haploinsufficiency is indicated as being mainly responsible for the 1p32-p31 chromosome deletion syndrome phenotype (OMIM 613735). Our patient also has a deleted FOXD3 of the FOX gene family of transcription factors, which plays an important role in neural crest cell growth and differentiation. As the murine FOXD3-/- model shows craniofacial anomalies and abnormal common carotid artery morphology, it can be hypothesised that FOXD3 is involved in the pathogenesis of the craniofacial and vascular defects observed in our patient. In support of our assumption, we found in the literature another patient with a syndromic form of MA who had a deletion involving another FOX gene (FOXC1). In addition to describing the clinical history of our patient, we have reviewed all of the available literature concerning other patients with a 1p32p31 deletion, including cases from the Decipher database, and we have also reviewed the genetic disorders associated with MA, which is a useful guide for the diagnosis of syndromic form of MA

Expanding phenotype of schimke immuno-osseous dysplasia: Congenital anomalies of the kidneys and of the urinary tract and alteration of nk cells

Schimke immuno-osseous dysplasia (SIOD) is a rare multisystemic disorder with a variable clinical expressivity caused by biallelic variants in SMARCAL1. A phenotype\u2013genotype correlation has been attempted and variable expressivity of biallelic SMARCAL1 variants may be associated with environmental and genetic disturbances of gene expression. We describe two siblings born from consanguineous parents with a diagnosis of SIOD revealed by whole exome sequencing (WES). Results: A homozygous missense variant in the SMARCAL1 gene (c.1682G>A; p.Arg561His) was identified in both patients. Despite carrying the same variant, the two patients showed substantial renal and immunological phenotypic differences. We describe features not previously associated with SIOD\u2014both patients had congenital anomalies of the kidneys and of the urinary tract and one of them succumbed to a classical type congenital mesoblastic nephroma. We performed an extensive characterization of the immunophenotype showing combined immunodeficiency characterized by a profound lymphopenia, lack of thymic output, defective IL-7R\u3b1 expression, and disturbed B plasma cells differentiation and immunoglobulin production in addition to an altered NK-cell phenotype and function. Conclusions: Overall, our results contribute to extending the phenotypic spectrum of features associated with SMARCAL1 mutations and to better characterizing the underlying immunologic disorder with critical implications for therapeutic and management strategies

Regional Precuneus Cortical Hyperexcitability in Alzheimer's Disease Patients

Objective: Neuronal excitation/inhibition (E/I) imbalance is a potential cause of neuronal network malfunctioning in Alzheimer's disease (AD), contributing to cognitive dysfunction. Here, we used a novel approach combining transcranial magnetic stimulation (TMS) and electroencephalography (EEG) to probe cortical excitability in different brain areas known to be directly involved in AD pathology. Methods: We performed TMS-EEG recordings targeting the left dorsolateral prefrontal cortex (l-DLPFC), the left posterior parietal cortex (l-PPC), and the precuneus (PC) in a large sample of patients with mild-to-moderate AD (n = 65) that were compared with a group of age-matched healthy controls (n = 21). Results: We found that patients with AD are characterized by a regional cortical hyperexcitability in the PC and, to some extent, in the frontal lobe, as measured by TMS-evoked potentials. Notably, cortical excitability assessed over the l-PPC was comparable between the 2 groups. Furthermore, we found that the individual level of PC excitability was associated with the level of cognitive impairment, as measured with Mini-Mental State Examination, and with corticospinal fluid levels of Aβ42 . Interpretation: Our data provide novel evidence that precuneus cortical hyperexcitability is a key feature of synaptic dysfunction in patients with AD. The current results point to the combined approach of TMS and EEG as a novel promising technique to measure hyperexcitability in patients with AD. This index could represent a useful biomarker to stage disease severity and evaluate response to novel therapies. ANN NEUROL 2022

Usefulness and limitations of comprehensive characterization of mRNA splicing profiles in the definition of the clinical relevance of BRCA1/2 variants of uncertain significance

Highly penetrant variants of BRCA1/2 genes are involved in hereditary predisposition to breast and ovarian cancer. The detection of pathogenic BRCA variants has a considerable clinical impact, allowing appropriate cancer-risk management. However, a major drawback is represented by the identification of variants of uncertain significance (VUS). Many VUS potentially affect mRNA splicing, making transcript analysis an essential step for the definition of their pathogenicity. Here, we characterize the impact on splicing of ten BRCA1/2 variants. Aberrant splicing patterns were demonstrated for eight variants whose alternative transcripts were fully characterized. Different events were observed, including exon skipping, intron retention, and usage of de novo and cryptic splice sites. Transcripts with premature stop codons or in-frame loss of functionally important residues were generated. Partial/complete splicing effect and quantitative contribution of different isoforms were assessed, leading to variant classification according to Evidence-based Network for the Interpretation of Mutant Alleles (ENIGMA) consortium guidelines. Two variants could be classified as pathogenic and two as likely benign, while due to a partial splicing effect, six variants remained of uncertain significance. The association with an undefined tumor risk justifies caution in recommending aggressive risk-reduction treatments, but prevents the possibility of receiving personalized therapies with potential beneficial effect. This indicates the need for applying additional approaches for the analysis of variants resistant to classification by gene transcript analyses