Prevaccination Glucose Time in Range Correlates With Antibody Response to SARS-CoV-2 Vaccine in Type 1 Diabetes.

CONTEXT: Poor glucose control has been associated with increased mortality in COVID-19 patients with type 1 diabetes (T1D). OBJECTIVE: This work aimed to assess the effect of prevaccination glucose control on antibody response to the SARS-CoV-2 vaccine BNT162b2 in T1D. METHODS: We studied 26 patients with T1D scheduled to receive 2 doses, 21 days apart, of BNT162b2, followed prospectively for 6 months with regular evaluation of SARS-CoV-2 antibodies and glucose control. Immunoglobulin G (IgG) to spike glycoprotein were assessed by enzyme-linked immunosorbent assay, and serum neutralization by a live SARS-CoV-2 assay (Vero E6 cells system). Glycated hemoglobin A1c (HbA1c) and continuous glucose monitoring (CGM), including time in range (TIR) and above range (TAR), were collected. The primary exposure and outcome measures were prevaccination glucose control, and antibody response after vaccination, respectively. RESULTS: Prevaccination HbA1c was unrelated to postvaccine spike IgG (r = -0.33; P = .14). Of note, the CGM profile collected during the 2 weeks preceding BNT162b2 administration correlated with postvaccine IgG response (TIR: r = 0.75; P = .02; TAR: r = -0.81; P = .008). Patients meeting the recommended prevaccination glucose targets of TIR (≥ 70%) and TAR (≤ 25%) developed stronger neutralizing antibody titers (P < .0001 and P = .008, respectively), regardless of HbA1c. Glucose control along the study time frame was also associated with IgG response during follow-up (TIR: r = 0.93; P < .0001; TAR: r = -0.84; P < .0001). CONCLUSION: In T1D, glucose profile during the 2 weeks preceding vaccination is associated with stronger spike antibody binding and neutralization, highlighting a role for well-controlled blood glucose in vaccination efficacy.

De Novo Transcriptome Sequencing of Olea europaea L. to Identify Genes Involved in the Development of the Pollen Tube.

In olive (Olea europaea L.), the processes controlling self-incompatibility are still unclear and the molecular basis underlying this process are still not fully characterized. In order to determine compatibility relationships, using next-generation sequencing techniques and a de novo transcriptome assembly strategy, we show that pollen tubes from different olive plants, grown in vitro in a medium containing its own pistil and in combination pollen/pistil from self-sterile and self-fertile cultivars, have a distinct gene expression profile and many of the differentially expressed sequences between the samples fall within gene families involved in the development of the pollen tube, such as lipase, carboxylesterase, pectinesterase, pectin methylesterase, and callose synthase. Moreover, different genes involved in signal transduction, transcription, and growth are overrepresented. The analysis also allowed us to identify members in actin and actin depolymerization factor and fibrin gene family and member of the Ca(2+) binding gene family related to the development and polarization of pollen apical tip. The whole transcriptomic analysis, through the identification of the differentially expressed transcripts set and an extended functional annotation analysis, will lead to a better understanding of the mechanisms of pollen germination and pollen tube growth in the olive.

A De novo Transcriptomic Approach to Identify Flavonoids and Anthocyanins "Switch-Off" in Olive (Olea europaea L.) Drupes at Different Stages of Maturation.

Highlights A de novo transcriptome reconstruction of olive drupes was performed in two genotypesGene expression was monitored during drupe development in two olive cultivarsTranscripts involved in flavonoid and anthocyanin pathways were analyzed in Cassanese and Leucocarpa cultivarsBoth cultivar and developmental stage impact gene expression in Olea europaea fruits. During ripening, the fruits of the olive tree (Olea europaea L.) undergo a progressive chromatic change characterized by the formation of a red-brown "spot" which gradually extends on the epidermis and in the innermost part of the mesocarp. This event finds an exception in the Leucocarpa cultivar, in which we observe a destabilized equilibrium between the metabolisms of chlorophyll and other pigments, particularly the anthocyanins whose switch-off during maturation promotes the white coloration of fruits. Despite its importance, genomic information on the olive tree is still lacking. Different RNA-seq libraries were generated from drupes of "Leucocarpa" and "Cassanese" olive genotypes, sampled at 100 and 130 days after flowering (DAF), and were used in order to identify transcripts involved in the main phenotypic changes of fruits during maturation and their corresponding expression patterns. A total of 103,359 transcripts were obtained and 3792 and 3064 were differentially expressed in "Leucocarpa" and "Cassanese" genotypes, respectively, during 100-130 DAF transition. Among them flavonoid and anthocyanin related transcripts such as phenylalanine ammonia lyase (PAL), cinnamate 4-hydroxylase (C4H), 4-coumarate-CoA ligase (4CL), chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), flavonol 3'-hydrogenase (F3'H), flavonol 3'5 '-hydrogenase (F3'5'H), flavonol synthase (FLS), dihydroflavonol 4-reductase (DFR), anthocyanidin synthase (ANS), UDP-glucose:anthocianidin: flavonoid glucosyltransferase (UFGT) were identified. These results contribute to reducing the current gap in information regarding metabolic processes, including those linked to fruit pigmentation in the olive.

Developmental stimuli and stress factors affect expression of ClGLP1, an emerging allergen-related gene in Citrus limon.

Germins and germin-like proteins (GLPs) constitute an ubiquitous family of plant proteins that seem to be involved in many developmental and stress related processes. A novel GLP cDNA was isolated from Citrus limon and structural features and genomic organization were investigated by in silico and Southern blots analysis. In lemon, the ClGLP1 encodes a 24.38 kDa which possesses a conserved motif of plant GLPs proteins. A phylogetic analysis mapped ClGLP1 as belonging to the GER3 subfamily into the GLP1 group of large GLP family. ClGLP1 was differentially expressed in the various organs and was highest in mature fruit. Moreover, expression in the fruit was tissue- and stage-related as well as dependent on agricultural practice (organic vs conventional). ClGLP1 transcripts increased during the transition from the green (180 days after blooming) to the yellow (240 days after blooming) mature fruit and were strongly enhanced in yellow mature fruit from organic compared with conventional culture. A sudden and systemic increase in ClGLP1 expression level was observed in leaves injured by wounding, together with an increase of endogenous H2O2 amount. Notably, an enhancement of H202 was observed in fruit peel during transition from green to yellow fruit stage. All together our data showed that ClGLP1 expression can be modulated in relation to both developmental stimuli and culture practices; evidence is also provided that through an oxidase activity this gene could play a role in fruit maturation as well as in stress responses.

Multi-probe in situ hybridization to whole mount Arabidopsis seedlings.

In situ RNA-RNA hybridization (ISH) is a molecular method for localization of gene transcripts at the cellular level and is widely used to provide spatial and temporal information regarding gene expression. However, standard protocols are complex and laborious to implement, restricting analysis to one or a few genes at any one time, each one observed on separate ISH preparations. Multi-probe whole-mount in situ hybridization is a powerful technique to compare the expression patterns of two or more genes simultaneously in the same tissue or organ. We describe for the first time in plants, the detection of three different mRNAs in a single fixed whole mount Arabidopsis seedling. A combination of bright fluorescent secondary antibodies was used for the detection of riboprobes differentially labeled by digoxigenin, biotin and fluorescein. The 3-D detection of each of the multiple fluorescent hybridization signals or in combination was obtained through confocal laser-scanning microscopy. The reliability of the method was tested in the root, using the PINFORMED (PIN) genes with non-overlapping temporal and spatial expression patterns. In the shoot, a class-I KNOTTED -like homeobox gene from Arabidopsis (KNAT1) with expression restricted to the shoot apical meristem was used in combination with ELONGATOR3 (ELO3) gene. In addition, the expression patterns of ELONGATOR complex gene (ELO2, ELO3) and HISTONE MONOUBIQUITINATION1 (HUB1) genes were analyzed in both shoot and root and a partial overlapping was observed. The whole procedure takes only 6 days.

Role of geranylgeranyl reductase gene in organ development and stress response in olive (Olea europaea) plants.

The NADPH-dependent geranylgeranyl reductase gene (OeCHLP) was characterised in olive (Olea europaea L.). OeCHLP catalyses the formation of carbon double bonds in the phytolic side chain of chlorophyll, tocopherols and plastoquinones and, therefore, is involved in metabolic pathways related to plant productivity and stress response, besides to nutritional value of its products. The nuclear OeCHLP encodes a deduced product of 51 kDa, which harbours a transit peptide for cytoplasm-to-chloroplast transport and a nicotinamide binding domain. Two estimated identical copies of gene are harboured per haploid genome of the cv. 'Carolea' used in the present study. Levels and cytological pattern of OeCHLP transcription were investigated by quantitative RT-PCR and in situ hybridisation. In line with the presence of ubiquitous tocopherols and/or chlorophyll, OeCHLP transcripts were present in various organs of plants. In leaves and fruits at different developmental stages, OeCHLP was differentially expressed in relation to their morpho-physiological features. An early and transient enhancement of gene transcription was detected in leaves of different age exposed to cold treatment (4°C), as well as in fruits mechanically wounded. Moreover, OeCHLP transcripts locally increased in specific cell domains of fruits severely damaged by the pathogen Bactrocera olea. Combined, these data show that OeCHLP expression early responds to biotic and abiotic stressful factors. Levels of tocopherols also increased in leaves exposed to cold conditions and fruits severely damaged by pathogen. We suggest that gene activity under stress condition could be related to tocopherol action.