Chemosensory protein 16 was identified in the hemolymph of Galleria mellonella as a protein with an amount increasing after oral infection with 10^3 CFU of Pseudomonas entomophila, and decreasing after infection with a higher dose (10^5 CFU) of bacteria. The expression of the CSP16 gene occurred in the fat body and in the gut and correlated with changes in the protein level in the hemolymph. The CSP16 protein inhibited P. entomophila growth in the concentration range from 0.15 to 6 nM. Additionally, the CSP16 protein showed bactericidal activity against P. entomophila, Bacillus thuringiensis, and Escherichia coli in the range of 2-18 μM, but only in the presence of protease inhibitors, otherwise it was degraded by extracellular proteases secreted by P. entomophila. We demonstrated that the bactericidal activity of CSP16 was related to its ability to perforate bacterial cellular membranes in a dose-dependent manner. The antimicrobial properties of this protein were also confirmed with the use of Atomic Force Microscopy, which showed significant changes in the topology of different bacterial cell surfaces. Finally, when CSP16 was injected in vivo into G. mellonella larvae one hour after infection with P. entomophila, more survivors were observed at particular time-points. Taking into account its immune properties and putative ability to bind bacteria-derived compounds, the possible function of CSP16 in the host-pathogen interaction is discussed.

L-proline (Pro) is a natural amino acid with known antioxidant and cryoprotectant activity. This study aimed to assess the impact of Pro supplementation in freezing medium on blastocyst survival and quality. In vitro fertilization (IVF) was conducted using oocytes collected from Korean cattle, and Day 7 blastocysts were cryopreserved through slow freezing. Optimal post-thaw blastocyst survival was determined by adding various Pro concentrations to the freezing medium. Additionally, the effect of Sucrose (Suc) alone or in conjunction with Pro was evaluated. To assess blastocyst quality, we analyzed reactive oxygen species (ROS) levels, apoptosis, and gene expression in blastocysts that survived 24 h after slow freezing-thawing. The hatching rate at 72 h was significantly higher in the 0.3 M Pro group than that in the 0 M group (p = 0.0466). The hatching rates at 48 and 72 h were significantly higher in the Pro group than in the Suc and Suc + Pro groups (48 h: Suc, p = 0.0037; Suc + Pro, p = 0.0052; 72 h: Suc, p = 0.0024; Suc + Pro, p = 0.0009). ROS levels and the apoptosis index were significantly lower in the Pro group than in the Suc group (p = 0.0099, and 0.0098, respectively). Furthermore, mRNA expression of HSPA1A was significantly lower in the Pro and Suc + Pro groups than in the Suc group (p = 0.0074, and p = 0.01174, respectively). Additionally, GCLC mRNA expression was significantly higher in the Pro group than in the Suc group (p = 0.0308). These findings indicate that Pro supplementation in a slow freezing medium enhances the viability and quality of embryos.

Oxidative stress and alterations in redox signalling have been implicated in the pathophysiology of myocardial infarction (MI). NADPH oxidase (Nox) is an important source of reactive oxygen species (ROS) in the infarcted myocardium. Alarmin S100A8/A9 amplifies acute myocardial inflammation in MI and has been shown to be a promising therapeutic target to improve cardiac function post-MI. We aimed to elucidate the underlying mechanisms linking S100A8/A9, oxidative stress and the inflammatory response in MI. MI was induced by permanent left coronary artery ligation in C57BL/6J mice, followed by treatment with the S100A8/A9 inhibitor ABR-238901 (30 mg/kg) or PBS for 3 days. The in-vivo experiments were complemented with mechanistic studies on cultured macrophages (Mac), important cellular effectors in MI. Compared to sham-operated animals, we detected significant increases in the Nox1, Nox2, Nox4 catalytic subunits at mRNA and protein levels, and NADPH-dependent ROS production in the left ventricle of MI mice. S100A8/A9 blockade prevented the up-regulation of Nox1/2/4 expression, reduced ROS formation, suppressed NF-kB activation and prevented NLRP3 inflammasome priming and activation, leading to reduced levels of active IL-1β. In-vitro, S100A8/A9 induced gene expression of Nox catalytic subtypes and NLRP3 in Mac in a TLR4-dependent and dose-dependent manner. These effects were counteracted by pharmacological inhibition of S100A8/9, TLR4, Nox1/4 and Nox2. In conclusion, we show that Nox upregulation and ROS formation triggered by S100A8/A9 contributes to NLRP3 inflammasome priming and increased IL-1β production in the infarcted myocardium. These mechanisms can be therapeutically targeted to prevent inflammatory and oxidant myocardial damage in acute MI.

Phaeodactylum tricornutum is a promising host for light-driven synthesis of heterologous proteins. However, the marine cold-water environment and alkaline-acidic pH shifts in the culture, necessitated by the diatom's growth requirements. In this study, we analyzed the influence of growth condition on maturation and dynamics of the yellow fluorescent protein (YFP) in episomal-transformant P. tricornutum. A mathematical model was developed to detect the parameters that affect biomass and YFP production. Optimized conditions increased YFP mean fluorescence intensity (MFI) per cell by 4.2-fold (3.6 ± 0.6 to 15.4 ± 1.1) and total protein levels in the culture by 1.8-fold (123 ± 4 to 219 ± 9 µg L-1), without affecting biomass. YFP stability studies in P. tricornutum showed that the ubiquitin-proteasome system contributes the degradation of the recombinant protein, whereas newly synthesized YFP remains stable for up to 12 h. This optimization provides insights into the fluorescent protein-based heterologous production in diatoms.

In this study, resistant variants of Salmonella enterica serovar Typhimurium SL1344 to different stressors were selected. In addition, a genetic and phenotypic study was performed to explore the mechanisms underlying the acquisition of resistance. We isolated 4 variants with increased stable resistance to acid, osmotic stress, high hydrostatic pressure (HHP) and Ultraviolet-C light (UV-C) after repeated rounds of exposure to these agents and outgrowth of survivors. A PEF-resistant variant (SL1344-RS), previously isolated by Sagarzazu et al. (2013), was also included in the analysis. The results indicated that the isolated variants showed resistance to at least one other agent. This increased resistance, in general terms, had a fitness cost in growth, and exerted a variable impact on virulence (mainly in cell adhesion capacity), increased antibiotic resistance but did not influence in biofilm formation capacity. Whole Genome Sequencing (WGS) analysis allowed us to identify the genetic changes responsible for these phenotypic differences, and revealed that in 3 out of the 5 variants (including SL1344-RS) a mutation was found in hnr gene, an anti-sigma factor that promotes RpoS proteolysis. Hence the expression of several rpoS-regulated genes was quantified and found higher in these variants. This increase in RpoS activity would explain the lower growth rates observed in these 3 variants, as it would lead to increased transcription of genes involved in growth arrest and resistance to various types of stress. However, further analysis of a set of 22 additional Salmonella strains obtained from different culture collections indicated that a direct relationship between RpoS activity and stress resistance might not exist within Salmonella.

Muscle disuse results in complex signaling alterations followed by structural and functional changes, such as atrophy, force decrease, and slow-to-fast fiber-type shift. Little is known about human skeletal muscle signaling alterations under long-term muscle disuse. In this study, we describe the effects of 21-day dry immersion on human postural soleus muscle. We performed both transcriptomic analysis and Western blots to describe the states of the key signaling pathways regulating soleus muscle fiber size, fiber type, and metabolism. Twenty-one-day dry immersion resulted in both slow-type and fast-type myofibers atrophy, downregulation of rRNA content, and mTOR signaling. Twenty-one-day dry immersion also leads to slow-to-fast fiber-type and gene expression shift, upregulation of p-eEF2, p-CaMKII, p-ACC content and downregulation of NFATc1 nuclear content. It also caused massive gene expression alterations associated with calcium signaling, cytoskeletal parameters, and downregulated mitochondrial signaling (including fusion, fission, and marker of mitochondrial density).NEW & NOTEWORTHY The main findings of our study are as follows: 1) The soleus slow fibers atrophy after 21-day dry immersion (DI) does not exceed that after 7-day DI; 2) The soleus ubiquitin ligases expression after 21-day DI returns to its initial level; 3) The soleus slow fibers atrophy after 21-day DI is accompanied by a mitochondrial apparatus structural markers decrease; 4) The soleus fibers signaling pathways restructuring process during 21-day DI is carried out in a complex manner.

Reduced skeletal loading and inactivity leads to muscle atrophy in humans and most mammals. By contrast, hibernating mammals demonstrate limited loss of skeletal muscle mass and strength by the end of winter after being physically inactive for several months. The present study objective was to detect any signs of muscle atrophy and restoration in arctic ground squirrel (AGS) skeletal muscles during the hibernation season. Quadriceps muscles of juvenile AGS males were collected 1-2 weeks before hibernation, and at 2, 6, 10-12 and 16-22 weeks after onset of hibernation during interbout arousal when body temperature returns to euthermic level. Muscle mass, fiber cross-sectional area (CSA) and fiber type composition were determined, as well as total and ribosomal RNA content, and expression of key genes involved in protein degradation. We found that muscle mass, CSA and fiber size distribution were not different between the groups (P > 0.05). No difference was detected in myofiber composition between the hibernation groups compared to pre-hibernation. Total RNA and ribosomal RNA content were not significantly different between the groups during hibernation. Transcript levels of ubiquitin E3-ligase FBXO32 (Atrogin-1, MAFbx) and autophagy related genes MAP1LC3A and BECN1 were not different between the hibernation and pre-hibernation groups. However, ubiquitin E3-ligase TRIM63 (MuRF-1) was significantly higher expressed at 2 weeks of hibernation compared to the other timepoints. These results, for the first time, show that AGS preserve muscles during hibernation season.

Fibroblasts are considered a key player in the wound healing process. Although this cellular family is constituted by several distinct subtypes, dermal fibroblasts are crucial thanks to their ability to secrete pro-regenerative growth factors, extracellular matrix (ECM) proteins and their immune and anti-inflammatory role. Sophorolipids (SL), sophorosides (SS) and glucolipids (G), mono-unsaturated (C18:1) or saturated (C18:0), glycolipids derived from microbial fermentation of wild type or engineered yeast Starmerella bombicola, constitute a novel sustainable class of bio-based chemicals with interesting physicochemical characteristics, which allow them to form soft diverse structures from hydrogels to vesicles, micelles or complex coacervates with potential interest in skin regeneration applications. In this study, we first tested the cytocompatibility of a broad set of molecules from this family on normal human dermal fibroblasts (NHDF). Our results show that, up to an upper threshold (0.1 % w/v), the microbial glycolipids (SL-C18:1, G-C18:1, SSbola-C18:1, SL-C18:0 and G-C18:0) under study were able to sustain cell growth. Furthermore, we selected the least cytotoxic glycolipids (SL-C18:1, SSbola-C18:1, SL-C18:0) to study their potential to promote wound healing by measuring the gene expression of several key skin regeneration markers (i.e. collagen, elastin, transforming growth factor β, fibroblast growth factor …) using qPCR. Unfortunately, none of these glycolipids modulated the gene expression of molecules involved in tissue repair. However, this study aims to encourage the community to test this novel class of molecules for novel high-end biomedical applications.

Biosurfactants prepared by microbial fermentation are natural amphiphiles of growing importance, with the goal of replacing synthetic surfactants in commercial formulations. However, their cytotoxicity profile is still poorly known, especially for new molecules like single-glucose lipids or bolaform sophorolipids. This wants to contribute to all those applications, which could be developed with biosurfactants in contact with the skin (cosmetics, wound healing). We test the cytotoxicity of five structurally-related molecules (C18:1 and C18:0 sophorolipids, C18:1 and C18:0 single-glucose lipids, C18:1 di-sophoroside) against normal human dermal fibroblasts (NHDF) and evaluate the metabolic activity of the least toxic among them. To the best of our knowledge, cytotoxicity of these molecules, and of microbial biosurfactants in general, was never tested against NHDF.

Fish are exposed to numerous stressors which negatively affect their immune response and increase infection susceptibility. The risk of bacterial infections results in the excessive and preventive use of antibiotics. Therefore, we aimed to study how antibiotic treatment and restraint stress will affect the stress response, microbiota composition, gut morphology, and inflammatory reaction in common carp. Both restraint stress and antibiotic treatment increased cortisol level. Moreover, antibiotics induced dysbiosis in fish gut, manifested by a decrease in the total abundance of bacteria, and a shift in bacteria diversity, including a reduced number of Aeromonas, Bacteroides, Barnesiellaceae, Cetobacterium and Shewanella and an increased abundance of Flavobacterium. To a lesser extent, stress modified gut microbiota, as it decreased bacteria number and slightly changed the microbiota composition by decreasing Cetobacterium abundance and increasing Vibrio abundance. Microbiota of the antibiotic-treated and stressed fish shifted from the beneficial bacterial genera - Cetobacterium and Bacteroides, to the increased presence of unfavorable bacteria such as Brevinema, Flavobacterium and Desulfovibrionaceae. Stress and antibiotic-induced changes in the gut microbiota were related to the changes in the gut morphology when the higher abundance of goblet and rodlet cells and increased secretion activity of goblet cells were observed. Moreover, up-regulation of the expression of genes encoding pro-inflammatory mediators and cytokines involved in the Th17 immune response was present in the gut of the antibiotic-treated and stressed fish. We conclude that in carp antibiotics and stress alter the abundance and composition of the microbiota and induce Th17-dependent inflammatory reaction in the gut. Moreover, our results strongly suggest the interplay of the stress axis and the brain-gut-microbiota axis.

Dopamine agonists, such as cabergoline (Cab), have demonstrated efficacy in restoring reproductive function in cases of hyperprolactinemia and hormonal dysregulation. This study investigates the long-term consequences of maternal Cab treatment on the reproductive phenotype of the progeny in a female transgenic (TG) mouse model with hyperprolactinemia and infertility due to human chorionic gonadotropin (hCG) β-subunit overexpression. The TG females that received Cab between weeks 3-4 of life exhibited a reversion of hyperprolactinemia and infertility, whereas WT females retained their fertility. When TG-cab- or WT-Cab-treated females were crossed with WT or TG males, respectively, their female TG offspring showed a reversal of precocious puberty, regularization of estrous cycles, fertility, and prevention of hyperprolactinemia and prolactinomas. Despite the persistent high LH/hCG bioactivity, the normalization of prolactin levels led to a reduction in ovarian luteinization markers and progesterone levels. The TG female pups born to either WT-cab- or TG-cab-treated females exhibited a normalized phenotype, thus suggesting that the effects were indeed due to maternal Cab administration, and not to the transgene. Cross-fostering experiments showed that the long-lasting programming effect of maternal Cab on offspring occurred during lactation because the TG female pups from non-treated WT female/TG male pregnancies, but nursed by Cab-treated females, were free from the altered TG phenotype. These results suggest that Cab treatment before pregnancy may have a multigenerational effect on the hypothalamic-pituitary-gonadal axis of the offspring, mediated during lactation. This highlights potential implications for generational health and clinical practices regarding the use of dopamine agonists during lactation.

Neuroinflammation is one of the main players in lesion expansion and locomotor deficits after spinal cord injury (SCI), thus treatments to control the inflammatory process emerge as novel therapeutic strategies. In this context, the anti-inflammatory effects of tetrahydrocannabinol (THC) and cannabidiol (CBD), the main phytocannabinoids of Cannabis sativa, are increasingly recognized. The aim of this work was to investigate the effects of a standardized Cannabis sativa extract (CSE), which is mainly composed by THC/CBD in equimolar concentration, on neuroinflammation, secondary damage and locomotor outcome after SCI in rats. After acute SCI, CSE therapy increased the number of non-inflammatory (arginase-1 positive) microglial cells in the epicenter of the lesion and decreased the number of pro-inflammatory ones (arginase-1 negative) in the epicenter and in the rostral and caudal regions of the lesion. CSE also reduced the number of reactive astrocytes in the grey matter of the rostral and caudal regions. These results are consistent with the downregulation of mRNAs of inflammatory mediators (IL-1β, TNFα, IL-6, C3) and the upregulation of anti-inflammatory markers (ARG-1, MRC). In the chronic phase, CSE treatment prevented cyst expansion and also increased the volume of spared grey and white matter. Regarding locomotor outcome, CSE-treated rats showed better locomotor scores (open field test), higher latency to fall (Rotarod test) and lower number of hindlimb foot misplacements (horizontal ladder walking test) than untreated injured rats. These results suggest that this standardized CSE offers a promising perspective for reducing acute neuroinflammation and promoting functional recovery after SCI.

Our current understanding of the pathogenesis of Ross River virus (RRV) infection has been derived from murine models, which do not reproduce clinical disease as experienced by infected humans and horses. This prompted us to establish more relevant host model systems to study host-virus interactions using ex vivo peripheral blood mononuclear cells (PBMCs) and in vitro primary synovial fibroblast and epidermal keratinocyte cultures. Transcriptomic analysis revealed that the expression of the transmembrane protein matrix remodelling associated 8 (mxra8), recently found to be responsible for RRV cell entry, was downregulated in all cell types when infected with RRV, compared to mock-infected controls. Potent antiviral and inflammatory responses were generated by both synovial fibroblasts and epidermal keratinocytes upon RRV infection. Upregulation of multiple genes, inducible by double-stranded RNA, together with upregulation of toll-like receptor (TLR) tlr-3, but not tlr-7, 8 and 9, suggests possible abortive replication of RRV in these cell types and potent antiviral mechanisms. This was corroborated by virus growth kinetic studies which indicated inefficient RRV replication in synovial fibroblasts and epidermal keratinocytes. Cellular metabolic flux studies on PBMCs and synovial fibroblasts showed that RRV infected cells had reduced mitochondrial function. In addition, compared to PBMCs of seronegative horses, an enhanced antiviral state and reduced inflammation related gene expression was seen in PBMCs of seropositive horses infected with RRV. Thus, despite potent antiviral and inflammatory responses via the interferon pathway exhibited in all cell types, restricting virus growth, mitochondria capacity and function of infected cells remained negatively impacted.

The rising sea surface temperatures driven by climate change cause thermal stress, leading to oxidative stress, metabolic disorders, and increased disease susceptibility, thereby impairing the physiological functions of fish. Therefore, understanding the adaptation mechanisms of fish to high temperatures is essential for mitigating the negative impacts of thermal stress on aquaculture productivity and fish health. In this study, Paralichthys olivaceus were subjected to high temperatures following pre-heating to evaluate the advantages of pre-stimulation prior to exposure to the critical temperature. The P. olivaceus were exposed to four groups; Acute (subjected to acute heat shock at 32 °C), AH-S (exposed to acquired heat shock at 28 °C followed by short recovery of 2 h and subsequent heat shock at 32 °C), AH-L (exposed to acquired heat shock at 28 °C followed by long recovery of 2 days and subsequent heat shock at 32 °C) and AH-SL (combined of AH-S and AH-L protocols). In terms of antioxidant response, mRNA expression (caspase 10, thioredoxin (Trx), superoxide dismutase (SOD), peroxiredoxin (Prx), glutathione-S-transferase (GST), and transferrin (TF)) and enzyme activities (SOD, CAT, and GST) were significantly upregulated in P. olivaceus pre-heated prior to high-temperature exposure (AH-S, AH-L, and AH-SL groups). In addition, the stress gene expressions such as heat shock protein 70 (HSP70), HSP60, HSP90, warm-temperature-acclimation-associated 65-kDa protein (Wap65-1), and glucose-regulated protein 78 (GRP78) was significantly upregulated in AH-S, AH-L and AH-SL groups. Pre-heating has been found to be effective in mitigating thermal stress, with the efficacy varying according to the differences in pre-heating methods.

The sea lice Caligus elongatus and Lepeophtheirus salmonis are both causing problems in salmonid aquaculture. Since the salmonid specialist L. salmonis represents the dominating problem, research on host-parasite interactions has focused on L. salmonis and Atlantic salmon (Salmo salar), while less is known for the generalist C. elongatus. As new knowledge can be found in the comparison between a specialist and a generalist, the present study compares the salmon immune responses and louse modulatory proteins between C. elongatus and L. salmonis. While the severity of skin lesions inflicted underneath both lice species appeared similar, C. elongatus seemed to be better at dampening inflammatory responses than L. salmonis. A comparison of exocrine gland genes encoding proteins with known effect at the host-parasite interface showed that C. elongatus express most of the genes previously identified in L. salmonis. Interestingly, three orthologues of the labial gland protein 3 (LGP3) known to induce cell death in salmonid immune cells were found. This expansion of the LGP3 gene might explain the limited influx of immune cells observed underneath C. elongatus, though yet unknown C. elongatus specific glandular proteins might also be at play. Despite the limited inflammatory response induced by adult C. elongatus, they provoke a forceful host anti-lice behaviour that is comparably less prominent in salmon infested with L. salmonis. Setule-like processes identified on the ventral surface of the C. elongatus marginal membrane might be of importance here, as could species specific behavioural differences or differences in the host modulatory proteins.

The antibiotic resistance of pathogenic bacteria is currently one of the major problems in medicine, and finding novel antibacterial agents is one of the most difficult tasks in the field of biomedical sciences. Studies on such tasks can be successful only if genetic and molecular mechanisms leading to antibiotic resistance/sensitivity are understood. Previous reports indicated that the bacterial protein Hfq, discovered as an RNA chaperone but subsequently demonstrated to play also other functions in cells, is involved in the mechanisms of the response of bacterial cells to antibiotics. Recently, it was found that Hfq dysfunction resulted in more effective accumulation of an antibiotic ciprofloxacin in Escherichia coli cells irrespective of the presence or absence of the AcrB efflux pump. However, small RNA-mediated impairment of expression of the ompF gene, which encodes a porin involved in antibiotics influx, reversed the effects of the absence of Hfq on the antibiotic accumulation. This led to the hypothesis that Hfq might influence ciprofloxacin accumulation in the manner independent on its RNA chaperone function, as this protein might also influence cellular membrane structure and functions. Here, we demonstrate that in ompC and ompF mutants of E. coli, accumulation of ciprofloxacin is significantly impaired in the absence of Hfq or its C-terminal domain. These results corroborate the above-mentioned hypothesis on a sRNA-independent mechanism of Hfq-mediated modulation of the antibiotic transmembrane transport. Since fluoroquinolones use both protein- and lipid-mediated pathways to cross the outer membrane, Hfq may influence both processes. This possibility will be discussed herein.

Frequent red tide outbreaks pose a serious threat to biodiversity and the safety of aquatic ecosystems. Bacillamides showed algicidal activity against algae. However, the low natural concentrations and their structural complexity hinder development of these molecules. Inspired by the natrual algicide Bacillamide A, a series of thiourea derivatives were synthesized. Bacillamide A derivative (3B) showed excellent algicidal activity against S. costatum (EC50 = 0.52 μg/mL) and P. minimum (EC50 = 2.99 μg/mL), respectively. In addition, it has low toxicity to mammals and is less toxic than copper sulfate. 3B treatment resulted in loss of algal cell integrity. It also decreased the Chlorophyll a content and Fv/fm of algal cells, while increasing the levels of malondialdehyde content, superoxide dismutase, and reactive oxygen. 3B also induced expression of the photosynthetic genes, including psaB, psbB, as well as the antioxidant genes SOD2 and CAT. This study demonstrates that Bacillamide A derivatives could provide a safer alternative for red tide algal management.

CYP97C1 as a haem-containing cytochrome P450 hydroxylase (P450-type) is important for carotene hydroxylation and xanthophyll biosynthesis. Research about this type of hydroxylase was mainly reported in several model plant species which have no specialized tissues accumulating massive carotenoids. The function of CYP97C1 in the horticultural plant, like carrots, was not fully studied. In this study, we focused on the role of DcCYP97C1 in carotenoid flux and colour formation in carrot. DcCYP97C1 was found highly expressed in the 'turning stage' of carrot taproot. Using stable transformation and CRISPR/Cas9-mediated gene knockout technology, DcCYP97C1 was confirmed the rate-limiting enzyme for lutein biosynthesis and important for taproot colour formation. Overexpression of DcCYP97C1 in an orange carrot KRD (Kurodagosun) resulted in five times overproduction of lutein accompanied by dramatic reduction of carotenes. Knockout of DcCYP97C1 in orange KRD and yellow carrot QTH (Qitouhuang) reduced all kinds of carotenoids including lutein, α-carotene and β-carotene reflecting the key role of DcCYP97C1 for total carotenoid accumulation in taproot 'turning stage'. Our study demonstrated that manipulation of DcCYP97C1 was sufficient to influence carotenoid flux, change carrot colour and for high lutein production. The uncovered role of DcCYP97C1 may be helpful for understanding plant carotenoid metabolism and breeding colourful carrot cultivars.

Water deficit negatively impacts crop productivity and quality. Plants face these challenges by adjusting biological processes and molecular functions according to the intensity and duration of the stress. The cultivated potato (Solanum tuberosum) is considered sensitive to water deficit, thus breeding efforts are needed to enhance its resilience. To capture novel functional information and gene regulatory networks, we carried out mass spectrometry-based proteomics in potato cell suspensions exposed to abrupt or stepwise osmotic stresses. Both forms of stress triggered significant alterations in protein expression, though with divergent response mechanisms. Stress response pathways orchestrated by key proteins enrolled in primary and secondary metabolism, antioxidant processes, transcriptional and translational machinery and chromatin organization were found in adapted cells. Target metabolites and reactive oxygen species levels were quantified to associate functional outcomes with the proteome study. Remarkably, we also showed that adapted cells tolerate an array of diverse conditions, including anoxia, salt and heat stress. Finally, the expression patterns of genes encoding selected differentially expressed proteins were investigated in potato plants subjected to either drought or salt stress. Collectively, our findings reveal the complex cellular strategies of osmotic stress adaptation, identifying new fundamental genes that could enhance potato resilience.

In social species, early social experience shapes the development of appropriate social behaviours during conspecific interactions referred to as social competence. However, the underlying neuronal mechanisms responsible for the acquisition of social competence are largely unknown. A key candidate to influence social competence is neuroplasticity, which functions to restructure neural networks in response to novel experiences or alterations of the environment. One important mediator of this restructuring is the neurotrophin BDNF, which is well conserved among vertebrates. We studied the highly social fish Neolamprologus pulcher, in which the impact of early social experience on social competence has been previously shown. We investigated experimentally how variation in the early social environment impacts markers of neuroplasticity by analysing the relative expression of the bdnf gene and its receptors p75NTR and TrkB across nodes of the social decision-making network. In fish raised in larger groups, bdnf and TrkB were upregulated in the anterior tuberal nucleus, compared to fish raised in smaller groups, while TrkB was downregulated and bdnf was upregulated in the lateral part of the dorsal telencephalon. In the preoptic area (POA), all three genes were upregulated in fish raised in large groups, suggesting that early social experiences might lead to changes of the neuronal connectivity in the POA. Our results highlight the importance of early social experience in programming the constitutive expression of neuroplasticity markers, suggesting that the effects of early social experience on social competence might be due to changes in neuroplasticity.

A short-term ivermectin (IVM) exposure method was newly established to demonstrate effects of sublethal concentrations of IVM on the wild-type fruit fly, Drosophila melanogaster. Using a conventional glass-vial contact approach, exposures to IVM (0.01 to 1000 ppm) for 12 h durations or less were selected to assess the downstream impacts of short-term IVM exposures (STIEs) on fruit flies. Under these conditions, all female flies produced significantly higher levels of reactive oxygen species and malondialdehydes in their ovaries. Additionally, females treated with IVM for 12 h under the STIE conditions exhibited significantly increased levels of DNA damages in their ovaries. Despite the negative impacts described above, the mean percent hatchability values obtained from the eggs oviposited by the IVM-exposed females were not statistically different when compared to the hatchability of the unexposed females. Two concentrations (1 and 10 ppm) of IVM were selected to determine transgenerational effects following short-term IVM exposures. F1, F2 and F8 flies exposed to IVM showed significantly delayed developments (2.5-3.2, 2.5-3.0, and 0.9-1.3 days delayed, respectively). F5, F11 and F17 females showed significantly delayed IVM-induced sluggish behaviors in the presence of lethal IVM (1 %, w/v). F18 females transgenerationally exposed to 1 ppm IVM exhibited significantly increased levels of Mrp1 (8.7-fold) and Cyp6g2 (5.9-fold) transcripts compared to unexposed flies. Comparatively, F18 females transgenerationally exposed to 10 ppm IVM showed significantly increased levels of Cyp9f2 (2.6-fold) transcripts. Current study clearly demonstrated the effects of sublethal IVM on parent and filial generations of fruit flies, providing an important step toward understanding development of IVM resistance under the STIE conditions.

Limb regeneration in the Mexican axolotl relies on the dedifferentiation of mature limb cells into blastema cells, which gain the ability to respond to patterning signals that guide tissue regeneration. While limb nerves are essential to make the blastema cells competent to pattern, the mechanisms remain unclear due to the complex and overlapping signals in amputated limbs. To overcome this challenge, we developed the Competency Accessory Limb Model (CALM), a simplified limb regeneration assay to study the induction and maintenance of patterning competency. Using CALM, here we show specific temporal windows during which cells acquire competency and associate this state with distinct H3K27me3 chromatin signatures. Furthermore, a combination of FGF and BMP signaling is sufficient to induce patterning competency in limb wound cells, and the ErBB signaling pathway is a downstream epigenetic target of these signals. These findings offer new insights into the molecular regulation of regenerative patterning.

In 2009, the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines established standards for the design, execution, and reporting of quantitative PCR (qPCR) in research. The expansion of qPCR into numerous new domains has driven the development of new reagents, methods, consumables, and instruments, requiring revisions to best practices that are tailored to the evolving complexities of contemporary qPCR applications.

Transparent, clear, and comprehensive description and reporting of all experimental details are necessary to ensure the repeatability and reproducibility of qPCR results. These revised MIQE guidelines reflect recent advances in qPCR technology, offering clear recommendations for sample handling, assay design, and validation, along with guidance on qPCR data analysis. Instrument manufacturers are encouraged to enable the export of raw data to facilitate thorough analyses and re-evaluation by manuscript reviewers and interested researchers. The guidelines emphasize that quantification cycle (Cq) values should be converted into efficiency-corrected target quantities and reported with prediction intervals, along with detection limits and dynamic ranges for each target, based on the chosen quantification method. Additionally, best practices for normalization and quality control are outlined and reporting requirements have been clarified and streamlined. The aim is to encourage researchers to provide all necessary information without undue burden, thereby promoting more rigorous and reproducible qPCR research.

Building on the collaborative efforts of an international team of researchers, we present updates, simplifications, and new recommendations to the original MIQE guidelines, designed to maintain their relevance and applicability in the context of emerging technologies and evolving qPCR applications.

Pancreatic ductal adenocarcinoma (PDAC) is the main and the deadliest form of pancreatic cancer. This is a major problem of public health since it will become the second leading cause of death by cancer in the next few years, mainly due to the lack of efficient therapies. Transient Receptor Potential Cation Channel Subfamily M Member 7 (TRPM7) protein, a cation channel fused with a serine/threonine kinase domain is overexpressed in PDAC and associated with a low survival. In this work, we aim to study the role of kinase domain on pancreatic cell fates by using a model of kinase domain deletion by CRISPR-Cas9. PANC-1 and MIA PaCa-2 PDAC cell lines were used and kinase domain was deleted by CRISPR-Cas9 strategy. Kinase domain deletion (ΔK) was validated by RT-qPCR and western blots. The effect of kinase domain deletion on channel function was studied by patch-clamp and Mn2+-quenching. The cell phenotype was studied by MTT and cell migration/invasion assays. Finally, the role of kinase domain was studied in vivo in xenografted mice. Here we show that TRPM7 kinase domain is required to maintain a mesenchymal phenotype in PDAC cells. We also demonstrated that TRPM7 and PAK1 interact in the same protein complexes. Moreover, TRPM7 kinase domain is required for carcinogenesis and cancer cell dissemination in vivo. Intriguingly, the role of TRPM7 kinase is cell specific and may depend on the KRAS oncogene mutation status. In conclusion, TRPM7 kinase domain is required to maintain a mesenchymal and aggressive phenotype in PDAC cells, and it could be a promising target against PDAC.

Rice grain chalkiness occurs when grains fill under heat stress, greatly reducing grain quality. But the effects of heat stress during grain filling on the subsequent development and adaptive traits remain to be elucidated. Here, we evaluated the effects of heat stress during parental grain filling on the thermotolerance of subsequent plants grown under heat stress after anthesis. Subsequent plants were grown from control (25 °C) and heat-exposed (30 °C) parental seeds under natural conditions until anthesis. Then plants were divided into three treatment groups-control [parental plants] - control [subsequent plants] (CC, 25 °C), control-heat (CH, 30 °C), and heat-heat (HH, 30 °C). Plants grown from heat-stressed seeds had thicker and shorter flag leaves, which delayed leaf senescence and improved photosynthesis under heat stress. HH plants also had significantly less chalkiness than those of CH plants. DNA methylation analysis revealed that heat stress during grain filling significantly hypomethylated promoters of starch biosynthesis genes and hypermethylated those of α-amylase genes. Consequently, HH plants had significantly higher expression of starch biosynthesis genes and suppressed expression of starch-degrading α-amylase genes than CH plants under heat stress. We propose that heat stress during grain filling induces adaptive transgenerational memory that allows subsequent plants to better cope with heat stress.

The global seawater temperature is expected to further rise in the following years. While species have historically adapted to climatic variations, the current pace of climate change may exceed their ability to adapt. The abnormally increased seawater temperatures occasionally lead to high mortalities of marine bivalve mollusks, threatening the productivity of aquaculture and the sustainability of wild populations. This study investigates the antioxidant and cell death mechanisms of the Mediterranean mussel Mytilus galloprovincialis during a 25-day exposure to temperatures of 24°C, 26°C, and 28°C, by analyzing the transcription of key genes and assessing the oxidative damage on days 1, 3, 12, and 25. In addition, individuals resilient (survived at 28°C until day 30) and susceptible (died early at 26°C and 28°C) to thermal stress were collected to investigate potential polymorphisms in associated genes. The results showed increased transcription of antioxidant genes at higher temperatures. Elevated pro-apoptotic indices were initially observed at 26°C and a higher mortality than at 28°C. However, final mortality was much higher at 28°C. At 26°C, mussels exhibited the highest oxidative damage and pro-apoptotic indices after 25 days. At 28°C, although oxidative damage occurred after 24 hours, survivors maintained a prolonged activated antioxidant defense and increased lc3b transcription, which likely contributed to the observed reduction of pro-apoptotic and oxidative damage metrics on day 25, compared to 26°C. Further, the coding sequences of catalase, intracellular Cu-Zn superoxide dismutase (Cu-Zn sod), and fas-associated protein with death domain (fadd) from heat-resilient and heat-susceptible mussels were analyzed. Based on statistical correlation of nucleotide and genotype frequencies with resilience phenotypes, two novel single nucleotide polymorphisms (SNPs) in Cu-Zn sod and one in fadd were detected, potentially correlating with thermal stress resilience. These findings offer valuable insights into the physiological and genetic adaptations of M. galloprovincialis to rising temperatures and highlight loci potentially linking to thermal resilience.

Bacterivorous soil protists can have positive impacts on plant performance, making them attractive targets for novel strategies to improve crop production. However, we generally lack the knowledge required to make informed choices regarding the protist species to be used or the optimal conditions for such amendments. Here, we examined how identity, diversity, and timing of inoculation of well-described protists impacted plant development and rhizosphere microbiome assembly. We first studied the impact of individual inoculation of six well-characterized protists on lettuce growth, with Cercomonas sp. S24D2 emerging as the strain with the largest impact on plant growth. In a second step, we created a three-protist species mixture inoculant by adding two protist species (Acanthamoeba sp. C13D2 and a heterolobosean isolate S18D10), based on differences in their feeding patterns. We then inoculated Cercomonas sp. either individually or in the protist mixture to lettuce plants 1 week before, simultaneously with, or 1 week after seedling transfer. We monitored plant growth and nutrient content, as well as impacts on the resident soil and rhizosphere microbiome composition. We found that early protist inoculation provided the greatest increase in aboveground biomass compared to the non-inoculated control. Single- and mixed-species inoculations had similar impacts on plant development and only minor impacts on prokaryotic community composition. While early inoculation seems to be the most promising timing for eliciting the positive effects of protist amendments, further, more systematic studies will be necessary to determine the conditions and ecological interactions that yield consistent and predictable improvements in plant performance.

The application of microorganisms, including bacterivorous soil protists, has been increasingly suggested as a sustainable agricultural approach. While positive impacts of the presence of predatory protists have been generally reported, the effects of the selected species and amendment conditions are largely unknown. Here, we examined how identity, diversity, and timing of inoculation of well-described protists impacted plant development and rhizosphere microbiome assembly. One species emerged as the one having the strongest impact in our specific system. This result highlights the importance of species selection for optimal outcome, but also suggests a huge potential in the barely investigated protist diversity for targeted application. Furthermore, the application of the inoculants before plant transfer showed the strongest effects on plants, providing some useful and new insights on the optimal time for such amendments.

Pulmonary hypertension of the newborn is a life-threatening disorder characterized by elevated pulmonary vascular resistance due to maladaptation of the pulmonary circulation after birth. The etiology and mechanisms underlying pulmonary hypertension of the newborn remain unclear, hindering the development of effective treatment. We hypothesize that perinatal chronic hypoxia upregulates microRNA-210, which is essential for suppression of pulmonary arterial spontaneous transient outward currents (STOCs), resulting in pulmonary hypertension of the newborn.

We tested this hypothesis in a large animal model of pregnant sheep and newborn lambs exposed to chronic hypoxia by comparing loss- versus gain-of-function of microRNA-210.

Chronic perinatal hypoxia increases pulmonary vascular resistance and pulmonary arterial pressure in newborn lambs. The effect was mainly mediated by hypoxia after birth in the newborn. Mechanistically, we showed a significant decrease in microRNA-210 in pulmonary arteries after birth, but newborn hypoxia abolished this birth-induced reduction. We found that microRNA-210 mimic suppressed STOCs in newborn pulmonary arteries, and knockdown of microRNA-210 by microRNA-210-LNA prevented the hypoxia-induced reduction of pulmonary arterial STOCs. In vivo loss-of-function and gain-of-function experiments reveal that microRNA-210 is essential in the hypoxia-induced suppression of pulmonary arterial STOCs, increased pulmonary vascular resistance, and pulmonary hypertension in newborn lambs. Mechanistically, microRNA-210 suppressed pulmonary arterial STOCs via downregulation of iron-sulfur cluster assembly enzyme and large-conductance Ca2+-activated K+ channels.

We provide explicit evidence that neonatal hypoxia increases microRNA-210 expression, which is essential for suppression of STOCs, resulting in pulmonary hypertension in newborn lambs. Our study reveals new insights into the mechanisms and clinically meaningful targets for treatment of pulmonary hypertension of the newborn.

Mitochondrial DNA (mtDNA) replication requires a steady supply of deoxyribonucleotides (dNTPs), synthesized de novo by ribonucleotide reductase (RNR). In nondividing cells, RNR consists of RRM1 and RRM2B subunits. Mutations in RRM2B cause mtDNA depletion syndrome, linked to muscle weakness, neurological decline, and early mortality. The impact of RRM2B deficiency on dNTP pools in nondividing tissues remains unclear. Using a mouse knockout model, we demonstrate that RRM2B deficiency selectively depletes dATP and dGTP, while dCTP and dTTP levels remain stable or increase. This depletion pattern resembles the effects of hydroxyurea, an inhibitor that reduces overall RNR activity. Mechanistically, we propose that the depletion of dATP and dGTP arises from their preferred degradation by the dNTPase SAMHD1 and the lower production rate of dATP by RNR. Identifying dATP and dGTP depletion as a hallmark of RRM2B deficiency provides insights for developing nucleoside bypass therapies to alleviate the effects of RRM2B mutations.

Thousands of nucleus-encoded chloroplast proteins are synthesized as precursors on cytosolic ribosomes and posttranslationally imported into chloroplasts. Cytosolic accumulation of unfolded chloroplast precursor proteins (e.g., under stress conditions) is hazardous to the cell. The global cellular responses and regulatory pathways involved in triggering appropriate responses are largely unknown. Here, by inducible and constitutive overexpression of ClpD-GFP to result in precursor protein overaccumulation, we present a comprehensive picture of multilevel reprogramming of gene expression in response to chloroplast precursor overaccumulation stress (cPOS), reveal a critical role of translational activation in the expression of cytosolic chaperones (heat-shock proteins, HSPs), and demonstrate that chloroplast-derived reactive oxygen species act as retrograde signal for the transcriptional activation of small HSPs. Furthermore, we reveal an important role of the chaperone ClpB1/HOT1 in maintaining cellular proteostasis upon cPOS. Together, our observations uncover a cytosolic heat shock-like response to cPOS and provide insights into the underlying molecular mechanisms.

Adverse childhood experiences (ACEs) alter brain development, leading to vulnerability for chronic pain, mental health disorders, and suicidality. These effects often emerge during adolescence. Importantly, ACEs can occur prenatally, including when exposed to in utero intimate partner violence (IPV) or postnatally as maternal neglect. Maternal social support has demonstrated promise in the mitigation of ACE-related deficits. Oxytocin, which has a role in social-bonding and stress regulation, serves as a suitable surrogate for social support in preclinical studies. Therefore, we aimed to explore the effects of oxytocin on alleviating social deficits, nociception, and epigenetic changes resulting from models that aimed to mimic the stress normally induced following exposure to two ACEs: IPV in utero and maternal neglect. During pregnancy, dams were randomly assigned to experience the model of IPV or a sham insult. Following birth, offspring from the IPV group underwent 10 days of maternal separation. Dams received three days of oxytocin therapy while nursing. In adolescence, half of the offspring underwent a plantar surgery to induce pain. Overall, in adolescence, rats exposed to the ACEs exhibited increased nociceptive sensitivity and aberrant social interactions, particularly among males, further suggesting that ACEs can increase an individual's risk for chronic pain. The ACEs changed gene expression related to social behaviour and neuroplasticity. Maternal oxytocin normalized pain, social, and gene changes, while oxytocin levels in offspring correlated with nociceptive sensitivity. Although ACEs have enduring consequences, the outcomes are modifiable, and oxytocin may be a robust and implementable therapeutic capable of attenuating early adversity.

The high mortality rate of breast cancer and the difficulties associated with therapeutic resistance, especially in cases where targeted treatments are unavailable, make it a serious threat to women's health. This study examines the relationship between three mature microRNAs (miRNAs) that are clustered together, namely miR- 23a, miR- 27a, and miR- 24-2, as well as their potential correlation with breast cancer.

We identified common gene targets of miR- 23a, miR- 27a, and miR- 24-2 using computational analysis. We also checked for the levels of miR- 23a, miR- 27a, and miR- 24-2 in 26 breast tumor tissues (with their matched control) as well as MCF7 and MDA-MB- 231 cell lines using qRT-PCR. Dual-luciferase reporter assay was conducted to validate the binding site of the microRNAs in their target gene. Western blot was performed to study the expression of various breast cancer related genes in the presence of the three microRNAs. In addition, the effect of microRNAs in cancer cell metastasis and cell division was carried out using invasion and cell cycle assay.

Computational analysis identified key genes, including GSK3β, NCOA1 and SP1, which are functionally linked to tumor progression and various other malignancies. All three microRNAs were found to be significantly downregulated in the breast cancer tissue samples in comparison to their respective controls. Kaplan-Meier plot analysis revealed that the expression levels of these genes and associated microRNAs correlates with breast cancer patient survival rates. Reduced SP1 and NCOA1 levels predicted a worse prognosis, but elevated levels of GSK3β were linked with decreased survival. Moreover, miR- 23a and miR- 24-2 specifically target GSK3β, potentially disrupting the Wnt/β-catenin pathway involved in breast cancer development. Functional tests showed that miR- 23a, miR- 27a and miR- 24-2 affect expression of EMT related genes, influencing cell invasion and migration, impacting ERK signaling and EMT, critical in the spread of breast cancer.

This study unlocks the potential of targeting the microRNA cluster as a therapeutic approach and emphasizes the complex regulatory roles of each individual members of the miR- 23a/27a/24-2 cluster in the pathogenesis of breast cancer.

Background: animal studies have shown that enteral stimuli play an important role in modulating gut-associated lymphoid tissue thus fasting may exert detrimental effects on such. Objective: the main objectives of this study were 1) to evaluate the effect of 3-day fasting on secretory immunoglobulin A (S-IgA) in parotid saliva and 2) to determine the levels of lymphotoxin (LT) transcription in peripheral blood mononuclear cells (PBMNC) from healthy volunteers as a proxy for studying GALT health. Methods: these adult volunteers had fasted for three days as part of a cultural ritual. Eleven volunteers (seven men and four women) with a median age of 43 (40-56) were included. Parotid saliva and blood samples were collected on day 0 (no fasting) and three days after fasting. Parotid saliva was obtained using a modified on-Crittenden device, and S-IgA was quantified using ELISA. Total RNA was isolated from peripheral blood mononuclear cells, and the level of lymphotoxin (LT) mRNA expression was determined by RT-PCR. Lipopolysaccharide (LPS), IL-10 (interleukin), tumor necrosis factor (TNF)α, body composition and other metabolic indicators were measured. Results: median BMI was 27.3 kg/m2 (24.9-29.1). After 3 days of fasting, there were no significant differences in S-IgA concentrations (p = 0.657), LT expression (p = 0.063), LPS (p = 0.182), nor IL-10 (p = 0.110). We found a statistical difference in TNFα levels (13.5 vs 11.3 pg/mL; p = 0.005) between the 0 and 3-day samples; TNFα decreased after fasting. Conclusion: further studies are needed to clarify the role of LT in the production of S-IgA and its relationship with nutritional status and inflammatory factors.

Cytokine-induced apoptosis inhibitor 1 (CIAPIN1) is a crucial anti-apoptotic protein; however, its role and associated molecular pathways in ferroptosis remain largely unexplored. This study aimed to investigate the effects of CIAPIN1 on ferroptosis in lipopolysaccharide (LPS)-induced podocytes and the associated underlying phenomenon.

In this study, we recruited 50 sepsis patients (aged 56.63 ± 10.33) with acute kidney injury (AKI), 50 sepsis patients without AKI, and 50 healthy controls. We established an in vitro model of LPS-induced MPC5 podocytes. RT-qPCR and Western blotting were used to evaluate mRNA and protein expression, respectively.

Serum CIAPIN1 is downregulated in patients with septic AKI and LPS-induced podocytes. CIAPIN1 overexpression (OE-CIAPIN1) attenuated cell proliferation and apoptosis in LPS-induced podocytes. OE-CIAPIN1 elevated phosphorylated phosphoinositide 3-kinase (p-PI3K; p85, Tyr458) and phosphorylated protein kinase B (p-Akt; Ser473) levels in LPS-induced podocytes. OE-CIAPIN1 significantly elevated synaptopodin mRNA levels and remarkably lowered desmin mRNA expression in MPC5 cells. In contrast, treatment with the PI3K/Akt pathway inhibitor, LY294002, reversed synaptopodin and desmin mRNA expression in MPC5 cells. Additionally, OE-CIAPIN1 reduced the malondialdehyde (MDA) content and Fe2 + concentration in the lysate of MPC5 cells, while elevating the MDA content and Fe2 + concentration by LY294002 treatment. Furthermore, OE-CIAPIN1 increased ferroptosis-related proteins, including solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4), in MPC5 cells, which was reversed by LY294002 treatment.

These results suggest that serum CIAPIN1 inhibits LPS-induced ferroptosis in podocytes by regulating the PI3K/AKT signaling pathway.

Grade IV astrocytoma, also referred to as glioblastoma (GBM), is the most common type of glioma, accounting for over 60% of all brain tumors. It is still a fatal illness in spite of years of investigation and does not currently have a treatment. Thus, scientists and medical professionals are constantly trying to understand the molecular processes and heterogeneity of GBM as well as looking for new ways to improve treatment results. Numerous studies have indicated that nanomaterials, and more especially nanoparticles, offer a great deal of potential for killing cancer cells; as a result, they are being considered as a potential alternative cancer treatment. Several studies have demonstrated that ZnO NPs have shown specific cytotoxicity against cancer cells while leaving normal cells unharmed. In this study we aim to synthesize sodium doped zinc oxide NPs using zingerone in an environmentally friendly manner to evaluate their cytotoxic effects on U87 GBM cell line and normal HEK cell line and investigate the occurrence of apoptosis via apoptosis assay by flowcytometry and gene expression study of TP53 and related genes to apoptosis and cell cycle regulation pathways. It was demonstrated that Na-doped ZnO NPs had a significant cytotoxic effect on U87 cells while having significantly less effect on normal HEK cells. Na-doped ZnO NPs eliminated cancerous cells through apoptosis induction and possibly cell cycle regulation via up-regulation of TP53, PTEN, BAX, P21 and down-regulation of Bcl2. The unique physicochemical properties of nanoparticles turn them into fascinating agents to treat GBM. Hence, the necessity of exploring the vast, yet unknown field of nanoparticles potentials cannot be over looked.

Ageing of adult males could be accelerated by both high mating/reproductive effort and fighting for mates. Testing the relative importance of these factors is challenging, however, because males that win fights also tend to have more mates. We used a 2 × 2 experimental design to test how a prolonged (9 week) period of either winning or losing fights, and either high or low reproductive effort (manipulating by varying access to females) interacts to affect male ageing and future reproduction allocation in the mosquitofish, Gambusia holbrooki. We measured telomere length and several life-history traits, including mating effort and ejaculates (sperm count and velocity). After 9 weeks, there were significant differences between winners and losers in their mating effort but not in their ejaculates. Males with a higher past reproductive effort (i.e., access to females) had significantly lower current mating effort and grew more slowly. Males with a higher past reproductive effort also had slower swimming sperm, but only if they were smaller than average in body size. Surprisingly, neither males with a higher past reproductive effort nor males that repeatedly lost fights had shorter telomeres. Our findings show that past social dynamics affect how males allocate resources to reproduction and somatic maintenance.

Decline of mitochondrial respiratory chain (mtRC) capacity is a hallmark of mitochondrial diseases. Patients with mtRC dysfunction often present reduced skeletal growth as a sign of premature cartilage degeneration and aging, but how metabolic adaptations contribute to this phenotype is poorly understood. Here we show that, in mice with impaired mtRC in cartilage, reductive/reverse TCA cycle segments are activated to produce metabolite-derived amino acids and stimulate biosynthesis processes by mechanistic target of rapamycin complex 1 (mTORC1) activation during a period of massive skeletal growth and biomass production. However, chronic hyperactivation of mTORC1 suppresses autophagy-mediated organelle recycling and disturbs extracellular matrix secretion to trigger chondrocytes death, which is ameliorated by targeting the reductive metabolism. These findings explain how a primarily beneficial metabolic adaptation response required to counterbalance the loss of mtRC function, eventually translates into profound cell death and cartilage tissue degeneration. The knowledge of these dysregulated key nutrient signaling pathways can be used to target skeletal aging in mitochondrial disease.

The development of biodegradable nanoparticles (NPs) for delivering anticancer drugs, such as letrozole (LTZ), offers a targeted approach for cancer therapy. In this study, we synthesized poly(ε-caprolactone)-co-poly(ethylene glycol) (PCL-co-PEG) and fabricated LTZ-loaded PCL-co-PEG NPs (LTZ-NPs) via emulsion-solvent evaporation. Folic acid (FA), a folate receptor-targeting molecule, was conjugated to the LTZ-loaded NPs (LTZ-FNPs) to enhance treatment efficacy against hormone receptor-positive breast cancer cells. Both NPs and FNPs exhibited a spherical morphology (60-90 nm), with FNPs showing higher drug entrapment efficiency and controlled release. LTZ release was minimal at physiological pH but increased in acidic, cancer-like environments, following the Korsmeyer-Peppas model, indicating a combination of Fickian and non-Fickian diffusion. In cytotoxicity assays, LTZ-FNPs exhibited higher toxicity against MCF-7 cells than LTZ-NPs. Controlled LTZ release altered gene expression, reducing B-cell leukemia/lymphoma 2 protein (Bcl2) and increasing caspase 8 (Casp8), promoting apoptosis. A shift to the SubG1 phase further confirmed enhanced LTZ-FNP-mediated cell death. Furthermore, p53 expression increased, while matrix metalloproteinase 9 (MMP-9) decreased, inhibiting cell invasion. This study introduces a biodegradable system with FA-functionalized, pH-sensitive NPs for the targeted and controlled delivery of LTZ. This approach holds great potential for selective, efficient treatment while minimizing systemic toxicity in breast cancer therapy.

Excess light induces chloroplast degradation in plants, leading to decreased photosynthetic efficiency and an albino leaf phenotype. However, the molecular mechanism underlying this process remains unclear, especially in perennial crops like tea plant. This study investigated the effects of relatively strong light (SL, 240 μmol m-2·s-1) on chloroplast ultrastructure and metabolites in the light-sensitive tea germplasm Nanchuan Dachashu (Camellia nanchuanica). Continuous exposure to SL resulted in abnormal chloroplast structure characterized by extensive vacuolation. SL also significantly decreased the levels of chlorophyll (-60.30 %), carotenoids (-88.29 %), free amino acids (-23.97 %), and caffeine (-41.15 %) compared to relatively weak light (WL, 15 μmol m-2·s-1). Transcriptome analysis and RT-qPCR revealed that the chloroplast vesiculation gene CsCV was significantly up-regulated under SL, with promoter analysis showing more light-responsive elements in CsCV compared to another light-responsive gene, CsNBR1. Overexpression of CsCV in Arabidopsis caused stunted growth and accelerated leaf senescence, with the most affected line showing decreases in chlorophyll and carotenoid contents of 24.97 % and 17.39 %, respectively. Conversely, silencing CsCV in tea plants using antisense oligodeoxynucleotides (asODNs) for 3 days increased chlorophyll and carotenoid levels by 15.98 % and 18.35 %, respectively. Bimolecular fluorescence complementation (BiFC) assays and in protein-protein docking simulations demonstrated that CsCV interacts with the photosystem proteins CsLhca1, CsLhcb4, and CsPsaL through its conserved C-terminal region, suggesting CsCV may trigger chloroplast degradation by destabilizing the photosynthetic apparatus under SL. These findings provide mechanistic insights into light-induced chloroplast degradation in tea plants and highlight CsCV as a potential target for improving crop stress tolerance.

Primary cilia are cell signalling and environment sensing organelles and have important roles during embryogenesis and homeostasis. We demonstrate glucocorticoid signalling is essential for normal cilia formation in mouse and human renal tubules. RNA sequencing of E18.5 kidneys from glucocorticoid receptor (GR) null mice identified significant reductions in key ciliogenesis-related genes including Ccp110, Cep97, Cep290 and Kif3a. Confocal microscopy reveals abnormal, stunted cilia on proximal tubules, podocytes, and collecting duct cells in mice with global or conditional deletion of GR. In contrast, activation of GR signalling with dexamethasone in human kidney organoids or mouse IMCD3 cells increases cilia length, an effect blocked by the GR antagonist RU486. Analysis of GR-null kidney extracts demonstrates reduced levels of pERK and SUFU identifying potential cell pathway crosstalk with GR signalling that coordinately regulate ciliogenesis in the renal tubule. Finally, dexamethasone reduces Aurora kinase A levels, a factor driving cilia disassembly and implicated in the pathogenesis of polycystic kidney disease.