Simplified Protocol for the Purification of Native Cas Nucleases for DNA-Free Genome Editing.

DNA-free genome editing by the direct delivery of CRISPR-associated nucleases has emerged as a promising technology due to its precision and reduced risk of off-target effects. However, existing purification protocols for native Cas proteins require the use of complex instrumentation, which limits their application. Here, we present a simplified protocol for the purification of native Cas9, Cas12RR and dCas9-VP64 nucleases optimized for DNA-free genome editing.

A Comparative Transcriptomic Study Reveals Temporal and Genotype-Specific Defense Responses to in Grapevine.

Grapevine ( L.), a globally significant crop, is highly susceptible to , the causative agent of gray mold disease. This study investigates transcriptomic responses to in tolerant and susceptible grapevine genotypes using RNA sequencing (RNA-seq).

Assembly of mTORC3 Involves Binding of ETV7 to Two Separate Sequences in the mTOR Kinase Domain.

mTOR plays a crucial role in cell growth by controlling ribosome biogenesis, metabolism, autophagy, mRNA translation, and cytoskeleton organization. It is a serine/threonine kinase that is part of two distinct extensively described protein complexes, mTORC1 and mTORC2. We have identified a rapamycin-resistant mTOR complex, called mTORC3, which is different from the canonical mTORC1 and mTORC2 complexes in that it does not contain the Raptor, Rictor, or mLST8 mTORC1/2 components.

On the Way to the Technological Development of Newly Selected Non- Yeasts Selected as Innovative Biocontrol Agents in Table Grapes.

Post-harvest decay of fresh table grapes causes considerable annual production losses. The main fungal agents of decay both in pre- and post-harvest are , spp., spp.

Distinct structural variants and repeat landscape shape the genomes of the ancient grapes Aglianico and Falanghina.

Mounting evidence recognizes structural variations (SVs) and repetitive DNA sequences as crucial players in shaping the existing grape phenotypic diversity at intra- and inter-species levels. To deepen our understanding on the abundance, diversity, and distribution of SVs and repetitive DNAs, including transposable elements (TEs) and tandemly repeated satellite DNA (satDNAs), we re-sequenced the genomes of the ancient grapes Aglianico and Falanghina. The analysis of large copy number variants (CNVs) detected candidate polymorphic genes that are involved in the enological features of these varieties.

Implementation of geriatric screening in the emergency department using the Consolidated Framework for Implementation Research.

Objective: Implementation of evidence-based care processes (EBP) into the emergency department (ED) is challenging and there are only a few studies of real-world use of theory-based implementation frameworks. We report final implementation results and sustainability of an EBP geriatric screening program in the ED using the Consolidated Framework for Implementation Research (CFIR).

Methods: The EBP involved nurses screening older patients for delirium (Delirium Triage Screen), fall risk (4-Stage Balance Test), and vulnerability (Identification of Seniors at Risk score) with subsequent appropriate referrals to physicians, therapy specialists, or social workers.

Development of mouse models with restricted HLA-B∗57:01 presentation for the study of flucloxacillin-driven T-cell activation and tolerance in liver injury.

Background: Flucloxacillin (FLX)-induced liver injury is immune-mediated and highly associated to HLA-B∗57:01 expression. Host factors leading to drug-induced liver injury are not yet well understood.

Objective: Characterize in vivo immune mechanisms determining the development of CD8 T cells reactive to FLX in animals expressing the risk human leukocyte antigen (HLA) allotype.

[Prognostic value of hospital hypoglycemia].

Background: Hospital hypoglycemia (HH) is an important problem since it is linked to hospital death, prolonged stay and higher costs, both in people with and without diabetes, in the critical area and general ward, especially in cases of severe hypoglycemia. The objective of our work was to compare hospital mortality, transfer to the critical area and hospitalization periods in adults with HH according to its severity.

Methods: We conducted a retrospective cohort of adults hospitalized with hypoglycemia at the Hospital Italiano de Buenos Aires between 2013 and 2018.

National opioid shortage is associated with increased use of ketamine infusion for analgosedation in the medical intensive care unit.

Objective: To assess the impact of the national shortage of injectable opioids during the winter of 2017-2018 on the use of ketamine infusion for analgosedation in the medical intensive care unit (MICU).

Design: A retrospective cohort study.

Setting: Single-center tertiary care MICU at The Ohio State University Wexner Medical Center.

VviAGL11 self-regulates and targets hormone- and secondary metabolism-related genes during seed development.

the Arabidopsis homolog, has been proposed to have a causative role in grapevine stenospermocarpy. An association between a mutation in the coding sequence (CDS) and the seedless phenotype was reported, however, no working mechanisms have been demonstrated yet. We performed a deep investigation of the full gene sequence in a collection of grapevine varieties belonging to several seedlessness classes that revealed three different promoter-CDS combinations.

Transcriptomic and methylation analysis of susceptible and tolerant grapevine genotypes following Plasmopara viticola infection.

Downy mildew, caused by the biotrophic oomycete Plasmopara viticola, is one of the most economically significant grapevine diseases worldwide. Current strategies to cope with this threat rely on the massive use of chemical compounds during each cultivation season. The economic costs and negative environmental impact associated with these applications increased the urge to search for sustainable strategies of disease control.

Novel and emerging biotechnological crop protection approaches.

Traditional breeding or genetically modified organisms (GMOs) have for a long time been the sole approaches to effectively cope with biotic and abiotic stresses and implement the quality traits of crops. However, emerging diseases as well as unpredictable climate changes affecting agriculture over the entire globe force scientists to find alternative solutions required to quickly overcome seasonal crises. In this review, we first focus on cisgenesis and genome editing as challenging biotechnological approaches for breeding crops more tolerant to biotic and abiotic stresses.

Native Vineyard Non- Yeasts Used for Biological Control of in Stored Table Grape.

Postharvest spoilage fungi, such as , are considered the main cause of losses of fresh fruit quality and vegetables during storage, distribution, and consumption. The current control strategy is the use of SO generator pads whose application is now largely under observation. A high quantity of SO can be deleterious for fresh fruits and vegetables and it is not allowed in organic agriculture.

Alterations in the HLA-B*57:01 Immunopeptidome by Flucloxacillin and Immunogenicity of Drug-Haptenated Peptides.

Neoantigen formation due to the interaction of drug molecules with human leukocyte antigen (HLA)-peptide complexes can lead to severe hypersensitivity reactions. Flucloxacillin (FLX), a β-lactam antibiotic for narrow-spectrum gram-positive bacterial infections, has been associated with severe immune-mediated drug-induced liver injury caused by an influx of T-lymphocytes targeting liver cells potentially recognizing drug-haptenated peptides in the context of HLA-B*57:01. To identify immunopeptidome changes that could lead to drug-driven immunogenicity, we used mass spectrometry to characterize the proteome and immunopeptidome of B-lymphoblastoid cells solely expressing HLA-B*57:01 as MHC-I molecules.

Color Stabilization of Apulian Red Wines through the Sequential Inoculation of and .

Mixed fermentation using and has gained attention in recent years due to their ability to modulate the qualitative parameters of enological interest, such as the color intensity and stability of wine. In this study, three of the most important red Apulian varieties were fermented through two pure inoculations of strains or the sequential inoculation of after 48 h from . The evolution of anthocyanin profiles and chromatic characteristics were determined in the produced wines at draining off and after 18 months of bottle aging in order to assess the impact of the different fermentation protocols on the potential color stabilization and shelf-life.

Correction to: SNP genotyping elucidates the genetic diversity of Magna Graecia grapevine germplasm and its historical origin and dissemination.

An amendment to this paper has been published and can be accessed via the original article.

Lessons Learned to Date on COVID-19 Hyperinflammatory Syndrome: Considerations for Interventions to Mitigate SARS-CoV-2 Viral Infection and Detrimental Hyperinflammation.

The first case of human transmission of SARS-CoV-2 was reported in China in December 2019. A few months later, this viral infection had spread worldwide and became a pandemic. The disease caused by SARS-CoV-2, termed COVID-19, is multifactorial and associated with both specific antiviral as well as inflammatory responses, the extent of which may determine why some individuals are asymptomatic while others develop serious complications.

Neural Networks for Cause of Hospitalization and Final Cause of Death Extraction from Discharge Summaries.

Determining the cause of death of hospitalized patients with cardiovascular disease is of the utmost importance. This is usually recorded in free text form. In this study we aimed to develop a series of supervised natural language processing algorithms to extract cardiovascular causes of hospitalization and final causes of death.

Capillaroscopy: a new application for the evaluation of vascular side-effects induced by chemotherapy.

Background: Cancer survivors are rising up, but this better survival is accompanied by possible treatments side-effects. In particular, cardiovascular effects are commonly reported, even if vascular damage is not necessarily connected to clinical manifestations. Periungual microcircle evaluation through capillaroscopy could identify asymptomatic patients with high risk of cerebro-cardio-vascular disease.

Microarray data and pathway analyses of peripheral blood mononuclear cells from healthy subjects after a three weeks grape-rich diet.

Using Human Gene Expression Microarrays (Agilent) technologies, we investigated changes of the level of gene expression in peripheral blood mononuclear cells of healthy subjects after 21 days of fresh table grape-rich diet and after an additional 28-day washout. Several hundreds of genes were differentially expressed after grape intake or after washout. The functional analysis of these genes detected significant changes in key processes such as inflammation and immunity, thrombosis, DNA and protein repair, autophagy and mitochondrial biogenesis.

Unraveling the Deep Genetic Architecture for Seedlessness in Grapevine and the Development and Validation of a New Set of Markers for VviAGL11-Based Gene-Assisted Selection.

Seedless inheritance has been considered a quasi-monogenic trait based on the VvAGL11 gene. An intragenic simple sequence repeat (SSR) marker, p3_VvAGL11, is currently used to opportunely discard seeded progeny, which represents up to 50% of seedlings to be established in the field. However, the rate of false positives remains significant, and this lack of accuracy might be due to a more complex genetic architecture, some intrinsic flaws of p3_VvAGL11, or potential recombination events between p3_VvAGL11 and the causal SNP located in the coding region.

FT-NIR Analysis of Intact Table Grape Berries to Understand Consumer Preference Driving Factors.

Fourier-transform near infrared spectroscopy (FT-NIR) is a technique used in the compositional and sensory analysis of foodstuffs. In this work, we have measured the main maturity parameters for grape (sugars and acids) using hundreds of intact berry samples to build models for the prediction of these parameters from berries of two very different varieties: "Victoria" and "Autumn Royal". Together with the chemical composition in terms of sugar and acidic content, we have carried out a sensory analysis on single berries.

The Glucosamine-derivative NAPA Suppresses MAPK Activation and Restores Collagen Deposition in Human Diploid Fibroblasts Challenged with Environmental Levels of UVB.

The ultraviolet (UV) component of solar radiation is the driving force of life on earth, but it can cause photoaging and skin cancer. In this study, we investigated the effects of the glucosamine-derivative 2-(N-Acetyl)-L-phenylalanylamido-2-deoxy-β-D-glucose (NAPA) on human primary fibroblasts (FBs) stimulated in vitro with environmental levels of UVB radiation. FBs were irradiated with 0.