Family Of Peptides Research Articles

B-015 Analytical Verification of MR-proADM Biomarker in Human EDTA Plasma on Immunoassay Analyzer LIAISON® XL of Diasorin

Abstract Background Adrenomedullin (ADM) is a potent vasodilator peptide of 52-aminoacid belonging to the calcitonin family of peptides. Midregional pro-ADM (MR-ProADM) is a precursor for ADM that circulates is in a 1:1 ratio with it, and proportionally represents its levels and activity. Increased ADM circulating concentrations have been described for several disease states, including dysfunction of the cardiovascular system and sepsis. However, reliable ADM quantification has been hampered by the short half-life, the existence of a binding protein, and its physical properties. Hence, quantificacion of MR-proADM represents a more suitable option. The biomarker MR-proADM has been shown to be an accurate prognostic marker for outcome prediction in patients with lower respiratory infections, sepsis, urinary tract infections, and kidney disease. The objective of this study was the verification of the metrological characteristics and the verification of the linearity of quantitative measurement range, as declared by the manufacturer, of the LIAISON® Brahms MR-proADMTM on Liaison® XL (DiaSorin). Methods The verification was performed with two levels of quality controls (QC) from the manufacturer (x̄L=1,19 nmol/L and x̄H=5,48 nmol/L), and residual EDTA plasma samples from patients with an active sepsis code. The evaluation included the assessment of: • Detection capability (including limit of quantification (LoQ) and limit of detection (LoD), measuring three samples at the manufacturer's stated concentration (0.21 nmol/L and 0.09 nmol/L, respectively) in duplicate within 4 runs). • Precision (CV, %) and accuracy (bias, %) within, measuring 10 replicates of each QC level; and between run, measuring 5 replicates for each QC level within 5 runs. • Linearity of the quantitative range of measurement (six intermediate samples prepared by mixing a high concentration sample- XH=9.64 nmol/L- and a low concentration sample, XL=0.59 nmol/L; measured in triplicate). All studies were performed in accordance with the recommendations of the Clinical Laboratory Standards Institute guidelines. Statistical analysis was performed using Microsoft Excel®, 2016. Results All results with diluted samples presented a signal (RLU) higher than that detected in samples without analyte, so the LoD was verified. The precision at the LoQ concentration was 26.7% (acceptance criteria was set as <20%). CV for within-run precision were 4.3% and 2.0% and for between-run precision 6.8% and 7.9% for low (L1) and high (L2) control level, respectively. The average bias was 3.60% for L1 and 4.23% for L2, meeting acceptance criteria (<8%). Linearity was confirmed between 0.59 and 9.64 nmol/L (R2 = 0.995). The biases between the expected and the obtained value were ≤7% through the quantitative range of measurement, except for the lowest concentration sample (close to the quantitation limit). Conclusions Our study showed good compliance with the metrological characteristics declared by the manufacturer for LoD, precision, bias and linearity range. Although the LoQ was not verified, it is outside the clinical decision value in septic patients (< 2.25 nmol/L) and it is not deemed a limitation for clinical practice. Additionally, the Liaison XL® platform permits us to configure an emergency protocol circuit (24/7) with a TAT of 35 min.

Targeted single cell expression profiling identifies integrators of sleep and metabolic state.

Animals modulate sleep in accordance with their internal and external environments. Metabolic cues are particularly potent regulators of sleep, allowing animals to alter their sleep timing and amount depending on food availability and foraging duration. The fruit fly, Drosophila melanogaster , suppresses sleep in response to acute food deprivation, presumably to forage for food. This process is dependent on a single pair of Lateral Horn Leucokinin (LHLK) neurons, that secrete the neuropeptide Leucokinin. These neurons signal to insulin producing cells and suppress sleep under periods of starvation. The identification of individual neurons that modulate sleep-metabolism interactions provides the opportunity to examine the cellular changes associated with sleep modulation. Here, we use single-cell sequencing of LHLK neurons to examine the transcriptional responses to starvation. We validate that a Patch-seq approach selectively isolates RNA from individual LHLK neurons. Single-cell CEL-Seq comparisons of LHLK neurons between fed and 24-hr starved flies identified 24 genes that are differentially expressed in accordance with starvation state. In total, 12 upregulated genes and 12 downregulated genes were identified. Gene-ontology analysis showed an enrichment for Attacins , a family of anti-microbial peptides, along with several transcripts with diverse roles in regulating cellular function. Targeted knockdown of differentially expressed genes identified multiple genes that function within LHLK neurons to regulate sleep-metabolism interactions. Functionally validated genes include an essential role for the E3 ubiquitin Ligase insomniac, the sorbitol dehydrogenase Sodh1, as well as AttacinC and AttacinB in starvation-induced sleep suppression. Taken together, these findings provide a pipeline for identifying novel regulators of sleep-metabolism interactions within individual neurons.

Marsupial immune protection is shaped by enhancer sharing and gene cluster duplication of cathelicidin antimicrobial peptides.

Marsupial neonates are born with immature immune systems, making them vulnerable to pathogens. While neonates receive maternal protection, they can also independently combat pathogens, though the mechanisms remain unknown. Using the sugar glider (Petaurus breviceps) as a model, we investigated immunological defense strategies of marsupial neonates. Cathelicidins, a family of antimicrobial peptides expanded in the genomes of marsupials, are highly expressed in developing neutrophils. Sugar glider cathelicidins reside in two genomic clusters and their coordinated expression is achieved by enhancer sharing within clusters and long-range physical interactions between clusters. These cathelicidins modulate immune responses and have potent antimicrobial effects, sufficient to provide protection in a mouse model of sepsis. Lastly, cathelicidins have a complex evolutionary history, where marsupials and monotremes are the only tetrapods that retained two cathelicidin clusters. Thus, cathelicidins are critical mediators of marsupial immunity, and their evolution reflects the life history-specific immunological needs of these animals.

A Peptide Strategy for Inhibiting Different Protein Aggregation Pathways.

Protein aggregation correlates with many human diseases. Protein aggregates differ in structure and shape. Strategies to develop effective aggregation inhibitors that reach the clinic failed so far. Here, we developed a family of peptides targeting early aggregation stages for both amorphous and fibrillar aggregates of proteins unrelated in sequence and structure. They act on dynamic precursors before mechanistic differentiation takes place. Using peptide arrays, we first identified peptides inhibiting the amorphous aggregation of a molten globular, aggregation-prone mutant of the Axin tumor suppressor. Optimization revealed that the peptides activity did not depend on their sequences but rather on their molecular determinants: a composition of 20-30 % flexible, 30-40 % aliphatic and 20-30 % aromatic residues, a hydrophobicity/hydrophilicity ratio close to 1, and an even distribution of residues of different nature throughout the sequence. The peptides also suppressed fibrillation of Tau, a disordered protein that forms amyloids in Alzheimer's disease, and slowed down that of Huntingtin Exon1, an amyloidogenic protein in Huntington's disease, both entirely unrelated to Axin. Our compounds thus target early stages of different aggregation mechanisms, inhibiting both amorphous and amyloid aggregation. Such cross-mechanistic, multi-targeting aggregation inhibitors may be lead compounds for developing drug candidates against various protein aggregation diseases.

Effect of Salinity and Salmon Pituitary Extract on the Expression of Reproduction and/or Salinity-Related Genes in the Pituitary Cells of Japaneses Eel.

Artificial sexual maturation of eel (Anguilla japonica) involves rearing in seawater and injecting salmon pituitary extract (SPE). The salinity of seawater and components of SPE influence hormonal activities of the eel pituitary, leading to gonad development. This study investigated the direct effects of salinity change and SPE treatment on the eel pituitary gland using primary cell cultures. Pituitary cells were cultured into four experimental groups: control culture (control), SPE-treated culture (SPE), NaCl-treated culture (NaCl) and NaCl+SPE treated culture (NaCl+SPE). We investigated the expression of genes presumably related to reproduction and/or salinity, including luteinizing hormone (LHβ), follicle stimulating hormone (FSHβ), progesterone receptor-like (pgrl), prolactin (PRL), dopamine receptor D4 (drd4), neuropeptide B/W receptor 2 (NPBWR2) and relaxin family peptide receptor 3-2b (rxfp3-2b). Gene expression analysis revealed significant upregulation of LHβ in SPE and NaCl+SPE groups compared to control and NaCl (p<0.05). FSHβ expression did not show any significant changes. PRL showed a significant decrease in the NaCl group (p<0.05). Pgrl, NPBWR2, drd4, and rxfp3-2b displayed the highest expression in the control group, with downregulation observed in all treatment groups (NaCl, SPE, and NaCl+SPE) (p<0.05). This study demonstrated the direct effects of salinity changes and SPE treatment on the eel pituitary. Results from this study also suggest that salinity change is necessary but work together with SPE to induce reproductive process, and that LHβ, pgrl, PRL, drd4, NPBWR2, and rxfp3-2b genes are obviously associated with reproduction and salinity changes in eels.

Acute Stress Affects the Relaxin/Insulin-Like Family Peptide Receptor 3 mRNA Expression in Brain of Pubertal Male Wistar Rats.

The current literature suggests that relaxin-3/relaxin/insulin-like family peptide receptor 3 (RLN-3/RXFP-3) system is involved in the pathophysiology of affective disorders because the results of anatomical and pharmacological studies have shown that the RLN-3 signaling pathway plays a role in modulating the stress response, anxiety, arousal, depression-like behavior, and neuroendocrine homeostasis. The risk of developing mental illnesses in adulthood is increased by exposure to stress in early periods of life. The available data indicate that puberty is especially characterized by the development of the neural system and emotionality and is a "stress-sensitive" period. The presented study assessed the short-term changes in the expression of RLN-3 and RXFP-3 mRNA in the stress-dependent brain regions in male pubertal Wistar rats that had been subjected to acute stress. Three stressors were applied from 42 to 44 postnatal days (first day: a single forced swim; second day: stress on an elevated platform that was repeated three times; third day: restraint stress three times). Anxiety (open field, elevated plus maze test) and anhedonic-like behavior (sucrose preference test) were estimated during these tests. The corticosterone (CORT) levels and blood morphology were estimated. We found that the RXFP-3 mRNA expression decreased in the brainstem, whereas it increased in the hypothalamus 72h after acute stress. These molecular changes were accompanied by the increased levels of CORT and anxiety-like behavior detected in the open field test that had been conducted earlier, that is, 24h after the stress procedure. These findings shed new light on the neurochemical changes that are involved in the compensatory response to adverse events in pubertal male rats and support other data that suggest a regulatory interplay between the RLN-3 pathway and the hypothalamus-pituitary-adrenal axis activity in the mechanisms of anxiety-like behavior.

Characterization of α-hairpinin in the genome of lima bean (Phaseolus lunatus)

Antimicrobial α-Hairpinin peptides are sources of bioactive molecules with significant structural diversity and various biological activities. Phaseolus lunatus, known as lima bean, is a plant from the Fabaceae family, native to Central and South America. It is the second most important legume agronomically and economically in tropical regions, after the common bean. Due to its importance, several omics studies have been recently conducted on this species. The research aimed to characterize the antimicrobial peptides of the α-hairpinin family in the lima bean genome by analyzing their physicochemical properties, in silico biological activities, and three-dimensional structure. To this end, α-hairpinin sequences were identified according to the cysteine pattern characteristic of this protein family. After target identification, a search for conserved domains, signal peptides, and subcellular localization was conducted. Molecular mass, isoelectric point, grand average of hydropathicity (GRAVY), and biological activities were also analyzed, and a three-dimensional model of the candidate was generated. Following the mining process, an α-hairpinin sequence from P. lunatus (PlHrp1) was identified. PlHrp1 consisted of 41 amino acids, was predicted to be in the extracellular region, had an isoelectric point of 6.28, a mass of 4.81 kDa, and a gravy of -1.078. The alignment revealed that PlHrp1 contained the 4 conserved cysteine residues. In silico activity analyses demonstrated that PlHrp1 exhibited antifungal and antibacterial activities, with no antiviral activity. The secondary structure displayed two antiparallel α-helices connected by a loop. The model described 94.4% of amino acid residues in favorable positions. This study provides a comprehensive analysis of α-hairpinin peptides in the Phaseolus lunatus genome, highlighting the presence of this peptide family in the lima bean species and offering insights into their potential.

Potential therapeutic use of dermaseptin S4 from the frog Phyllomedusa sauvagii and its derivatives against bacterial pathogens in fish.

Dermaseptins are one of the main families of antimicrobial peptides (AMPs) derived from the skin secretions of Hylidae frogs. Among them, dermaseptin S4 (DS4) is characterized by its broad-spectrum of activity against bacteria, protozoa, and fungi. In this study, the physicochemical properties of the native peptide DS4 (1-28) and two derivatives [DS4 (1-28)a and DS4 (1-26)a] isolated from the skin of the frog Phyllomedusa sauvagii were investigated and their antimicrobial properties against two marine pathogenic bacteria (Vibrio harveyi and Vibrio anguillarum) were examined. The results indicate that the peptide DS4 (1-26)a has high-antibacterial activity against the tested strains and low-hemolytic activity (<30% lysis at the highest tested concentration of 100 µg/mL) compared to the other two peptides tested. In addition, all three peptides affect the membrane and cell wall integrity of both pathogenic bacteria, causing leakage of cell contents, with DS4 (1-26)a having the most severe impact. These skills were corroborated by transmission electron microscopy and by the variation of cations in their binding sites due to the effects caused by the AMPs. These results suggest that DS4 and its derivatives, in particular the truncated and amidated peptide DS4 (1-26)a could be effective in the treatment of infections caused by these marine pathogenic bacteria. Future studies are required to validate the use of DS4 in vivo for the prevention of bacterial diseases in fish.

Rational design of self-assembling ultrashort peptides for the shape- and size-tunable synthesis of metal nanostructures.

Peptides have attracted great interest as platforms for the design of nanocomposite hydrogels due to their distinct bioactivity, biofunctionality and biocompatibility. Previously, we have reported on a family of peptides that self-assembled to form stabilised three-dimensional hydrogel networks, displaying potent antimicrobial activity. In this paper, we report on the use of these hydrogelator sequences and their analogues as stabilisers and growth controllers to synthesise anisotropic gold nanoparticles (AuNPs) of different sizes and shapes. In particular, hollow spherical nanoparticles were obtained for HG2.81-AuNPs, whereas hexagonal nanoparticles were observed for TOH_1N-AuNPs and PentaOH-AuNPs in their respective hydrogel networks. The PentaOH-AuNPs' hydrogel exhibited excellent results with high antimicrobial potency against Staphylococcus aureus and Pseudomonas aeruginosa ATCC 27853 and negligible cytotoxicity. On the other hand, TOH_1N-AuNPs showed no antibacterial activity and no cytotoxicity, demonstrating the versatility of these peptides. This work gives credence towards the development of these materials towards further applications such as in tissue culture technology and wound dressing materials.

Genome-wide identification of phytosulfokine (PSK) peptide family reveals TaPSK1 gene involved in grain development of wheat (Triticum aestivum L.)

BackgroundPhytosulfokine (PSK) functions as a plant peptide growth factor that plays important and diverse roles in plant development and stress responses. Nevertheless, PSKs have not been systematically analyzed in wheat.ResultsA genome-wide comparative analysis of PSK genes in wheat was conducted and 15 TaPSKs were identified and divided into four subgroups in the wheat genome based on sequence similarity. The examination of motif compositions of TaPSK genes revealed the presence of the YIYTQ signature motif in the C-terminus of all TaPSK polypeptide precursors, with a highly conserved feature across different species. Exogenous application of TaPSK peptide promoted root growth in wheat. Quantitative RT-PCR analysis revealed that the TaPSKs exhibited preferential or tissue-specific expression patterns in wheat. In particular, three homologs of the TaPSK1 genes were specifically expressed in grains, with the strongest expression observed in the developing grains at 15 days after anthesis. Compared with wild type, transgenic rice lines overexpressing TaPSK1-A exhibited larger grain size and higher thousand-grain weight. The promoter region and genomic sequence of the wheat TaPSK1-A gene were cloned. Sequence polymorphism showed that five single-nucleotide polymorphisms (SNPs) were identified in the promoter region of TaPSK1-A. A Kompetitive Allele-Specific PCR (KASP) marker was developed for TaPSK1-A based on –806 bp SNP (C/T transition), and two haplotypes, TaPSK1-A-HapI and TaPSK1-A-HapII were detected in 260 wheat accessions collected from different regions. The expression of TaPSK1-A, promoter activity, and thousand-grain weight (TGW) in the TaPSK1-A-HapII haplotype were higher than those in the TaPSK1-A-HapI haplotype. Furthermore, yeast one-hybrid assays revealed the binding of TaNF-YB1 and TaERF39 to the promoter regions of the TaPSK1-A gene, and TaMADS29 could bind to the promoters of TaPSK1-B and TaPSK1-D genes.ConclusionsComparative genome-wide analysis of TaPSK peptide family revealed that the TaPSK1 gene is involved in wheat grain development, and the developed TaPSK1-A-KASP marker could be utilized for marker-assisted selection breeding of wheat.Graphical

Structural Classification Insights Into the Plant Defensive Peptides.

This study presents a structural phylogenetic analysis of plant defensive peptides, revealing their evolutionary relationships, structural diversification, and functional adaptations. Utilizing a robust dataset comprising both experimental and predicted structures sourced from the RCSB Protein Data Bank and AlphaFold DB, we constructed a detailed phylogenetic tree to elucidate the distinct evolutionary paths of plant defensive peptide families. Our findings showcase the evolutionary intricacies of defensive peptides, highlighting their diversity and the conservation of key structural motifs critical to their antimicrobial or defensive functions. The results also underscore the adaptive significance of defensive peptides in plant evolution, highlighting their roles in responding to ecological pressures and pathogen interactions.

Novel BRICHOS-related defensin-like antimicrobial peptide from the marine polychaeta &lt;i&gt;Arenicola marina&lt;/i&gt;

To date, polychaetes remain a poorly studied class of invertebrate animals in the context of clarification of their immune system functioning and, in particular, of antimicrobial peptides (AMPs) biodiversity. AMPs, also known as host defense peptides, play a key role in host protection from various pathogens and regulation of the species composition of symbiotic microbes. The study of biosynthesis of AMPs in polychaetes has revealed an interesting pattern, namely so-called BRICHOS domain in the precursor proteins of a number of such peptides. The conserved structure of this domain allows to perform a bioinformatic search for AMP precursors in polychaete transcriptomes. In this work, we found and studied a new BRICHOS-associated AMP from the lugworm Arenicola marina, which represents a structural family of defensin-like peptides stabilized by four disulfide bonds, not previously identified in marine worms. The peptide, designated as AmBRI-44a, contained 44 amino acid residues and was obtained by heterologous expression in Escherichia coli. AmBRI-44a was shown to have a specific activity against a narrow spectrum of Gram-positive bacteria and did not exhibit pronounced cytotoxic effects on eukaryotic cell line HEK293T. A potential mechanism of the antibacterial action of this peptide may be associated with inhibition of bacterial cell wall biosynthesis, as indicated by genetic and phenotypic analysis of selected AmBRI-44a-resistant bacteria Bacillus licheniformis B-511. The results obtained allow us to consider the novel peptide AmBRI-44a as a candidate compound for the development of an antibiotic agent that could potentially be effective in the treatment of infectious diseases mediated by multidrug-resistant Gram-positive bacteria.

Exploring the Pharmacological Effects of Bioactive Peptides on Human Nervous Disorders: A Comprehensive Review.

A family of peptides known as bioactive peptides has unique physiological properties and may be used to improve human health and prevent illness. Because bioactive peptides impact the immunological, endocrine, neurological, and cardiovascular systems, they have drawn a lot of interest from researchers. According to recent studies, bioactive peptides have a lot to offer in the treatment of inflammation, neuronal regeneration, localized ischemia, and the blood-brain barrier. It investigates various peptide moieties, including antioxidative properties, immune response modulation, and increased blood-brain barrier permeability. It also looks at how well they work as therapeutic candidates and finds promising peptide-based strategies for better outcomes. Furthermore, it underscores the need for further studies to support their clinical utility and suggests that results from such investigations will enhance our understanding of the pathophysiology of these conditions. In order to understand recent advances in BPs and to plan future research, academic researchers and industrial partners will find this review article to be a helpful resource.

Biotechnological Production of the Recombinant Two-Component Lantibiotic Lichenicidin in a Bacterial Expression System

Objective: Lantibiotics are a family of bacterial antimicrobial peptides synthesized by ribosomes, that undergo post-translational modification to form lanthionine (Lan) and methyllanthionine (MeLan) residues. Lantibiotics are currently considered promising agents for combating antibiotic-resistant bacterial infections. This paper presents a biotechnological method for obtaining two components of the lantibiotic lichenicidin from Bacillus licheniformis B-511, Lchα and Lchβ. Such a system has the potential to facilitate the production of not only lichenicidin, but also other lantibiotics, including two-component ones, and also to enable the study of their biosynthesis and the activity and substrate specificity of modifying enzymes. Methods: The developed system is based on heterologous coexpression of the genes of Lchα and Lchβ precursors with the genes of their corresponding modifying enzymes in the cytoplasm of Escherichia coli BL21(DE3). Subsequent steps included immobilized metal affinity chromatography of the His-tagged hybrid peptide under denaturing conditions, cyanogen bromide cleavage in acidic medium, and final purification using reverse-phase HPLC. Results and Discussion: The system was employed for the expression and purification of lantibiotics, resulting in the successful isolation of the β-component of lichenicidin in high yield (approximately 4 mg/L of culture). This purified beta component exhibited structural and functional characteristics comparable to its natural counterpart, which was purified from the natural producer. However, the yield of the mature α-component of lichenicidin in such a system was significantly lower. Conclusions: The work presents a biotechnological method for obtaining recombinant two-component lantibiotic lichenicidin, which has proven to be particularly effective in the case of Lchβ. The developed method can also be applied to the production of other promising lantibiotics and their further research.

Functional expression and characterization of CAPA receptor in the digestive tract and life stages of Drosophila suzukii, and differential activities with insect PRXamide peptides.

Spotted-wing drosophila, Drosophila suzukii (Matsumura), is an invasive vinegar fly that is a major threat to the small fruits industries globally. Insect capa genes encode multiple neuropeptides, including CAPA-periviscerokinin (CAPA-PVK) peptides, that are specifically known to cause diuresis or anti-diuresis in various organisms. Here we identified and characterized a corresponding G protein-coupled receptor (GPCR) of the D. suzukii CAPA-PVK peptides: CAPA receptor (CAPA-R). To better characterize the behavior of D. suzukii CAPA-R, we used insect cell-based functional expression assays to evaluate responses of CAPA-RagainstD. suzukii CAPA-PVKs, CAPA-PVKs from five species in Insecta, one species from Mollusca, modified CAPA-PVK peptides, and some PRXamide family peptides: pyrokinin (PK), diapause hormone (DH), and ecdysis-triggering hormone (ETH). Functional studies revealed that the D. suzukii CAPA-R is strongly activated by both of its own natural D. suzukii CAPA-PVKs, and interestingly, it was strongly activated by other CAPA-PVK peptides from Frankliniella occidentallis (Thysanoptera), Solenopsis invicta (Hymenoptera), Helicoverpa zea (Lepidoptera) and Plutella xylostella (Lepidoptera). However, D. suzukii CAPA-R was not activated by Mollusca CAPA-PVK or the other PRXamide peptides. Gene expression analyses showed that the CAPA-R was highly expressed in the Malpighian tubules and moderately in hindgut compared to other digestive organs or the rest of body, supporting diuretic/antidiuretic functionality. When compared across life stages of D. suzukii, expression of CAPA-R was approximately 1.5x greater in the third instar than the other stages and minimally detected in the eggs, 4-day old pupae and 3-day old adults. Our results functionally characterized the D. suzukii CAPA-R and a few short peptides were identified as potential biological targets to exploit the CAPA-R for D. suzukii management.

A comprehensive characterization and expression profiling of defensin family peptides in Arabidopsis thaliana with a focus on their abiotic stress-specific transcriptional modulation

In addition to defensins, plants possess an array of defensin-like peptides that share many of their characteristics, as well as a role in plant’s innate immunity. Their involvement in the response to pathogens is well-known but the contribution in the plant response to abiotic stimuli is not fully understood. We have undertaken an in silico analysis to characterize all defensin family genes hitherto found in Arabidopsis, including genes encoding for defensin-like peptides, by detecting several peptides as candidates for further studies aiming to decipher specific responses to biotic and abiotic stresses, as well as to their crosstalk. We performed several analyses, including co-expression and cis-regulatory elements analyses, using transcriptomic data obtained from the ARS database, which integrates more than 20,000 Arabidopsis RNA-seq libraries.In silico analysis showed that jasmonates and ABA, together with transcription factors belonging to WRKY and AP2/EREBP families, modulate defensin and defensin-like gene expression. Indeed, the analysis performed in this study allowed to extract and organize omics data, which finally supported the inducible nature of defensins under both abiotic and biotic stresses. Moreover, in vivo expression analyses confirmed the heat and drought responsiveness of PDF1.4, ATTI1, PDF1.1, DEFL 206, defensin family genes selected for being upregulated by several abiotic conditions, at transcriptional level. Finally, the co-expression analysis provided information on other biological processes that may be correlated to the defensin induction, such as maintaining ROS homeostasis. Combining the comprehensive analysis of different transcriptional datasets with the integration of in vivo analyses emerged as a robust methodological approach to assess the proposed multi-stress responsive nature of defensin family genes.

Natural fermentation of fresh Jasmine flowers inspired the development of a biotechnological ingredient with global anti-ageing properties.

This study focused on the development of a new-to-world ingredient harnessing the natural potential of fresh Jasminum grandiflorum flowers to self-ferment by its phytobiome revealing flower content. Analytical investigations were conducted to highlight specific phytocompounds generated during the natural fermentation of flowers in comparison to a conventional extraction. The synergy with another extraction technology maximized the generation of biocompounds for an interesting efficacy. Jasmine extract was elaborated by combining two patented technologies: the phytofermentology™, inspired by plant-microorganisms interaction and designed to develop ingredients obtained by natural fermentation of the vegetal using its own phytobiota; and the PSR™ technology allowing the extraction of bioactive phytocompounds such as small RNAs from plants. Analytical investigations of Jasmine extract highlighted uniqueness and richness of the phytocompound profiles, such as organics acids and phenolic compounds, markers of fermentation only obtained after phytofermentology in comparison to conventional extraction. Jasmine extract has the particularity to contain jasmintides, flower small peptides belonging to the family of cysteine-rich peptides (CRPs). Antioxidant and global anti-ageing properties were investigated in cell-free assays demonstrating interesting results: about 20% scavenging of free radicals from 0.5% of Jasmine extract and protection from DNA damage of 26% in comparison to a stressed control. Phytofermentology™ technology combined with PSR™ technology, meant to be respectful of the environment, allowed to development of biofunctionals very close to nature with a unique analytical signature as Jasmine extract, using the potential of fresh flowers phytobiota to self-ferment. The efficacy of the ingredient on global antioxidation and anti-ageing via hyaluronidase/tyrosinase inhibitions was highlighted by cell-free evaluation assays. Further and complementary studies should be conducted to confirm the bioefficacy of this ingredient with invitro / exvivo assays.

Genome-Wide Association Analyses and Population Verification Highlight the Potential Genetic Basis of Horned Morphology during Polled Selection in Tibetan Sheep.

The types and morphology of sheep horns have been extensively researched, yet the genetic foundation underlying the emergence of diverse horn characteristics during the breeding of polled Tibetan sheep has remained elusive. Genome-wide association analysis (GWAS) was performed on 103 subtypes (normal large horn, scurs, and polled) differentiated from G2 (offspring (G2) of parent (G1) of polled) of the polled core herd. Six single nucleotide polymorphisms (SNPs) located on chromosome 10 of the relaxin family peptide receptor 2 (RXFP2) gene exhibited positive correlations with horn length, horn base circumference, and horn base interval. Furthermore, in genotyping 382 G2 individuals, significant variations were observed for each specific horn type. Three additional mutations were identified near the target SNP upstream of the amplification product. Finally, the RXFP2-specific haplotype associated with the horned trait effectively maintained horn length, horn base circumference, and horn base interval in Tibetan sheep, as confirmed by population validation of nine loci in a sample size of 1125 individuals. The present study offers novel insights into the genetic differentiation of the horned type during improvement breeding and evolution, thereby establishing a robust theoretical foundation for polled Tibetan sheep breeding and providing valuable guidance for practical production.

Characterization of corazonin signaling in a molluscan model species, Lymnaea stagnalis

In recent years, new concepts have emerged regarding the nomenclature, functions, and relationships of different peptide families of the gonadotropin-releasing hormone (GnRH) superfamily. One of the main driving forces for this originated from the emerging evidence that neuropeptides previously called molluscan GnRH are multifunctional and should be classified as corazonin (CRZ). However, research articles still appear that use incorrect nomenclature and attribute the same function to molluscan CRZs as vertebrate GnRHs. The aim of the present study was to further support the recent interpretation of the origin and function of the GnRH superfamily. Towards this goal, we report the characterization of CRZ signaling system in the molluscan model species, the great pond snail (Lymnaea stagnalis). We detected a CRZ-receptor-like sequence (Lym-CRZR) by homology-searching in the Lymnaea transcriptomes and the deduced amino acid sequence showed high sequence similarity to GnRH receptors and CRZ receptors. Molecular phylogenetic tree analysis demonstrated that Lym-CRZR is included in the cluster of molluscan CRZRs. Lym-CRZR transiently transfected into HEK293 cells was found to be localized at the plasma membrane, confirming that it functions as a membrane receptor, like other G protein-coupled receptors. The signaling assays revealed that the previously identified Lym-CRZ neuropeptide stimulated intracellular Ca2+ mobilization in a dose-dependent manner, but not cyclic AMP production, in HEK293 cells transfected with Lym-CRZR. Finally, we demonstrated a wide tissue distribution of Lym-CRZR. These results suggest that Lym-CRZ is a multifunctional peptide and provide further insights into the evolution of the GnRH neuropeptide superfamily. The present study also supports the notion that previously termed molluscan “GnRH” should be classified as “CRZ”.

Upgrading Mitochondria-Targeting Peptide-Based Nanocomplexes for Zebrafish In Vivo Compatibility Assays.

The lack of effective delivery systems has slowed the development of mitochondrial gene therapy. Delivery systems based on cell-penetrating peptides (CPPs) like the WRAP (tryptophan and arginine-rich peptide) family conjugated with a mitochondrial targeting sequence (MTS) have emerged as adequate carriers to mediate gene expression into the mitochondria. In this work, we performed the PEGylation of WRAP/pDNA nanocomplexes and compared them with previously analyzed nanocomplexes such as (KH)9/pDNA and CpMTP/pDNA. All nanocomplexes exhibited nearly homogeneous sizes between 100 and 350 nm in different environments. The developed complexes were biocompatible and hemocompatible to both human astrocytes and lung smooth muscle cells, ensuring in vivo safety. The nanocomplexes displayed mitochondria targeting ability, as through transfection they preferentially accumulate into the mitochondria of astrocytes and muscle cells to the detriment of cytosol and lysosomes. Moreover, the transfection of these cells with MTS-CPP/pDNA complexes produced significant levels of mitochondrial protein ND1, highlighting their efficient role as gene delivery carriers toward mitochondria. The positive obtained data pave the way for in vivo research. Using confocal microscopy, the cellular internalization capacity of these nanocomplexes in the zebrafish embryo model was assessed. The peptide-based nanocomplexes were easily internalized into zebrafish embryos, do not cause harmful or toxic effects, and do not affect zebrafish's normal development and growth. These promising results indicate that MTS-CPP complexes are stable nanosystems capable of internalizing in vivo models and do not present associated toxicity. This work, even at an early stage, offers good prospects for continued in vivo zebrafish research to evaluate the performance of nanocomplexes for mitochondrial gene therapy.