Molecular Stress Response Research Articles

Effects of commercial beverages on the neurobehavioral motility of Caenorhabditis elegans.

To study the effects of different types of commercially available drinks/beverages on neurobehavior using the model organism C. elegans, and critically review their potential health hazards. Eighteen kinds of beverages from the supermarket were randomly selected and grouped into seven categories namely functional beverage, tea beverage, plant protein beverage, fruit juice beverage, dairy beverage, carbonated beverage and coffee beverage. The pH value, specific gravity and osmotic pressure were also examined. The L4 stage N2 worms were exposed to different concentration of tested beverages (0, 62.5, 125, 250 and 500 µL/mL) for 24 h to measure the survival rate and locomotory behavior such as head thrashing, body bending as well as pharyngeal pumping. All the 18 beverages tested did not induce any visible lethal effects in the nematodes. However, exposure to different types of tested beverages exhibited different effects on the behavioral ability of C. elegans: (1) sports functional beverage and herbal tea drink accelerated the head thrashing and body bending of nematodes when compared to the control group (P < 0.05). (2) The vibration frequency of the pharyngeal pump of nematodes was significantly accelerated after treated with three plant protein beverages (almond milk, coconut milk and milk tea) and dairy products A and B (P < 0.05), and decelerated after treatment with other tested beverages. (3) Carbonated beverage significantly inhibits the head thrashing, body bending and pharyngeal pumping vibration (P < 0.05). Our results indicate that 18 kinds of popular beverages in the market have different influence on the neurobehavior in C. elegans, which may be related to their different components or properties. Further research would be required to conduct a systematic analysis of the effect of beverages by appropriate kinds, taking into consideration other endpoints such as reproduction, lifespan and molecular stress response, etc., and to elucidate the mechanism for its potential health hazards.

Editorial: High Throughput Omics Technologies for Understanding Molecular Stress Response and Devising Mitigation Actions

Editorial: High Throughput Omics Technologies for Understanding Molecular Stress Response and Devising Mitigation Actions

Simulated winter warming negatively impacts survival of Antarctica's only endemic insect

Abstract Antarctic winters are challenging for terrestrial invertebrates, and species that live there have specialised adaptations to conserve energy and protect against cold injury in the winter. However, rapidly occurring climate change in these regions will increase the unpredictability of winter conditions, and there is currently a dearth of knowledge on how the highly adapted invertebrates of Antarctica will respond to changes in winter temperatures. We evaluated the response of larvae of the Antarctic midge, Belgica antarctica, to simulated winters at three ecologically relevant mean temperature scenarios: warm (−1°C), normal (−3°C) and cold (−5°C). Within each scenario, larvae were placed into three distinct habitat types in which they are commonly observed (decaying organic matter, living moss, and Prasiola crispa algae). Following the simulated overwintering period, a range of physiological outcomes were measured, namely survival, locomotor activity, tissue damage, energy store levels and molecular stress responses. Survival, energy stores and locomotor activity were significantly lower following the Warm overwintering environment than at lower temperatures, but tissue damage and heat shock protein expression (a proxy for protein damage) did not significantly differ between the three temperatures. Survival was also significantly lower in larvae overwintered in Prasiola crispa algae, although the underlying mechanism is unclear. Heat shock proteins were expressed least in larvae overwintering in living moss, suggesting it is less stressful to overwinter in this substrate, perhaps due to a more defined structure affording less direct contact with ice. Our results demonstrate that a realistic 2°C increase in winter microhabitat temperature reduces survival and causes energy deficits that have implications for subsequent development and reproduction. While our Warm winter scenario was close to the range of observed overwintering temperatures for this species, warmer winters are expected to become more common in response to climate change. Conversely, if climate change reduces the length of winter, some of the negative consequences of winter warming may be attenuated, so it will be important to consider this factor in future studies. Nonetheless, our results indicate that winter warming could negatively impact cold‐adapted insects such as the Antarctic midge. Read the free Plain Language Summary for this article on the Journal blog.

Does previous exposure to extreme precipitation regimes result in acclimated grassland communities?

Climate change will likely increase weather persistence in the mid-latitudes, resulting in precipitation regimes (PR) with longer dry and wet periods compared to historic averages. This could affect terrestrial ecosystems substantially through the increased occurrence of repeated, prolonged drought and water logging conditions. Climate history is an important determinant of ecosystem responses to consecutive environmental extremes, through direct damage, community restructuring as well as morphological and physiological acclimation in species or individuals. However, it is unclear how community restructuring and individual metabolic acclimation effects interact to determine ecosystem responses to subsequent climate extremes. Here, we investigated, if and how, differences in exposure to extreme or historically normal PR induced long-lasting (i.e. legacy) effects at the level of community (e.g., species composition), plant (e.g., biomass), and molecular composition (e.g., sugars, lipids, stress markers). Experimental grassland communities were exposed to long (extreme) or short (historically normal) dry/wet cycles in year 1 (Y1), followed by exposure to an identical PR or the opposite PR in year 2 (Y2). Results indicate that exposure to extreme PR in Y1, reduced diversity but induced apparent acclimation effects in all climate scenarios, stimulating biomass (higher productivity and structural sugar content) in Y2. In contrast, plants pre-exposed to normal PR, showed more activated stress responses (higher proline and antioxidants) under extreme PR in Y2. Overall, Y1 acclimation effects were strongest in the dominant grasses, indicating comparatively high phenotypical plasticity. However, Y2 drought intensity also correlated with grass productivity and structural sugar findings, suggesting that responses to short-term soil water deficits contributed to the observed patterns. Interactions between different legacy effects are discussed. We conclude that more extreme PR will likely alter diversity in the short-to midterm and select for acclimated grassland communities with increased productivity and attenuated molecular stress responses under future climate regimes.

Ocular Surface Infection Mediated Molecular Stress Responses: A Review.

Infection mediated ocular surface stress responses are activated as early defense mechanisms in response to host cell damage. Integrated stress responses initiate the host response to different types of infections and modulate the transcription of key genes and translation of proteins. The crosstalk between host and pathogen results in profound alterations in cellular and molecular homeostasis triggering specific stress responses in the infected tissues. The amplitude and variations of such responses are partly responsible for the disease severity and clinical sequelae. Understanding the etiology and pathogenesis of ocular infections is important for early diagnosis and effective treatment. This review considers the molecular status of infection mediated ocular surface stress responses which may shed light on the importance of the host stress-signaling pathways. In this review, we collated literature on the molecular studies of all ocular surface infections and summarize the results from such studies systematically. Identification of important mediators involved in the crosstalk between the stress response and activation of diverse signaling molecules in host ocular surface infection may provide novel molecular targets for maintaining the cellular homeostasis during infection. These targets can be then explored and validated for diagnostic and therapeutic purposes.

Nematostella vectensis exhibits an enhanced molecular stress response upon co-exposure to highly weathered oil and surface UV radiation

Crude oil released into the environment undergoes weathering processes that gradually change its composition and toxicity. Co-exposure to petroleum mixtures and other stressors, including ultraviolet (UV) radiation, may lead to synergistic effects and increased toxicity. Laboratory studies should consider these factors when testing the effects of oil exposure on aquatic organisms. Here, we study transcriptomic responses of the estuarine sea anemone Nematostella vectensis to naturally weathered oil, with or without co-exposure to environmental levels of UV radiation. We find that co-exposure greatly enhances the response. We use bioinformatic analyses to identify molecular pathways implicated in this response, which suggest phototoxicity and oxidative damage as mechanisms for the enhanced stress response. Nematostella's stress response shares similarities with the vertebrate oxidative stress response, implying deep conservation of certain stress pathways in animals. We show that exposure to weathered oil along with surface-level UV exposure has substantial physiological consequences in a model cnidarian.

Cover Image, Volume 78, Issue 3

The cover image is based on the Research Article Selectivity and molecular stress responses to classical and botanical acaricides in the predatory mite Phytoseiulus persimilis Athias‐Henriot (Acari: Phytoseiidae) by Sabina A Bajda et al., https://doi.org/10.1002/ps.6747.image

Selectivity and molecular stress responses to classical and botanical acaricides in the predatory mite Phytoseiulus persimilis Athias-Henriot (Acari: Phytoseiidae).

Acaricide application remains an integral component of integrated pest management (IPM) for the two-spotted spider mite Tetranychus urticae. Species and strains of phytoseiid predatory mites vary significantly in their response to acaricides. For the success of IPM, it is imperative to identify the determinants of selectivity and molecular stress responses of acaricides in predatory mites. The three classical acaricides bifenazate, cyflumetofen, and fenbutatin oxide did not affect the survival and fecundity of Phytoseiulus persimilis regardless of the route of exposure. Selectivity of the orange oil and terpenoid blend-based botanical acaricides was low via a combination of direct exposure, acaricide-laced diet, and residual exposure but improved when limiting exposure only to diet. To gain insights into the molecular stress responses, the transcriptome of P. persimilis was assembled. Subsequent gene expression analysis of predatory mites orally exposed to fenbutatin oxide and orange oil yielded only a limited xenobiotic stress response. In contrast, P. persimilis exhibited target-site resistance mutations, including I260M in SdhB, I1017M in CHS1, and kdr and super-kdr in VGSC. Extending the screen using available Phytoseiidae sequences uncovered I136T, S141F in cytb, G119S in AChE, and A2083V in ACC, well-known target-sites of acaricides. Selectivity of the tested botanical acaricides to P. persimilis was low but could be enhanced by restricting exposure to a single route. Differential gene expression analysis did not show a robust induced stress response after sublethal exposure. In contrast, this study uncovered target-site mutations that may help to explain the physiological selectivity of several classical acaricides to phytoseiid predators.

The Interaction of Osmotic and Heavy Metal Stress in C. elegans

Cellular stress is an ever-present aspect of aging and a primary driver of many common age-associated diseases such as cancer, diabetes, or neurodegenerative diseases. As we age, stress-induced damage accumulates over time, along with reduced efficacy of stress response pathways at combatting such damage. Molecular stress response pathways are well studied in the context of individual stressors, but there is a lack of understanding of how these responses change when multiple stressors are encountered at the same time. The goal of our work is to explore the impact of multiple simultaneous stressors on health and survival, and to investigate the underlying molecular pathways involved. To accomplish this, we utilize the nematode Caenorhabditis elegans to monitor lifespan changes in response to various stressors. We simultaneously exposed C. elegans to high concentrations of sodium chloride and cadmium chloride, known to induce osmotic and heavy metal stress, respectively. We found that lifespan is drastically decreased by the combined stress, significantly more so than the reduction in lifespan caused by either individual stress. Our results show that glycerol levels, which are normally increased in response to osmotic stress, are significantly lowered when the two stresses are combined compared to levels detected for osmotic stress alone. This suggests that the presence of cadmium may sensitize worms to sodium and other osmotic stressors by blunting cells’ ability to mount an appropriate molecular response. In ongoing work, we will continue to dissect the mechanisms through which cadmium influences glycerol production and other aspects of osmotic stress response.

Effects of plant growth-promoting rhizobacteria on the molecular responses of maize under drought and heat stresses: A review

Drought and heat are major environmental stresses that continually influence plant growth and development. Under field conditions, these stresses occur more frequently in combination than alone, which magnifies corresponding detrimental effects on the growth and productivity of agriculturally important crops. Plant responses to such abiotic stresses are quite complex and manifested in a range of developmental, molecular, and physiological modifications that lead either to stress sensitivity or tolerance/resistance. Maize ( Zea mays L.) is known for its sensitivity to abiotic stresses, which often results in substantial loss in crop productivity. Bioaugmentation with plant growth-promoting rhizobacteria (PGPR) has the potential to mitigate the adverse effects of drought and heat stresses on plants. Hence, this is considered a promising and eco-friendly strategy to ensure sustainable and long-term maize production under adverse climatic conditions. These microorganisms possess various plant growth-promoting (PGP) characteristics that can induce drought and heat tolerance in maize plants by directly or indirectly influencing molecular, metabolic, and physiological stress responses of plants. This review aims to assess the current knowledge regarding the ability of PGPR to induce drought and heat stress tolerance in maize plants. Furthermore, the drought and heat stress-induced expression of drought and heat stress response genes for this crop is discussed with the mechanisms through which PGPR alter maize stress response gene expression.

Interactive effects of ZnO nanoparticles and temperature on molecular and cellular stress responses of the blue mussel Mytilus edulis

Temperature is an important abiotic factor that modulates all aspects of ectotherm physiology, including sensitivity to pollutants. Nanoparticles are emerging pollutants in coastal environments, and their potential to cause toxicity in marine organisms is a cause for concern. Here we studied the interactive effects of temperature (including seasonal and experimental warming) on sublethal toxicity of ZnO nanoparticles (nano-ZnO) in a model marine bivalve, the blue mussel Mytilus edulis. Molecular markers were used to assess the pollutant-induced cellular stress responses in the gills and the digestive gland of mussels exposed for 21 days to 10 μg l−1 and 100 μg l−1 of nano-ZnO or dissolved Zn under different temperature regimes including ambient temperature (10 °C and 15 °C in winter and summer, respectively) or experimental warming (+5 °C). Exposure to high concentration (100 μg l−1) of nano-ZnO caused oxidative injury to proteins and lipids and induced a marked apoptotic response indicated by increased transcript levels of apoptosis-related genes p53, caspase 3 and the MAPK pathway (JNK and p38) and decreased mRNA expression of anti-apoptotic Bcl-2. No significant induction of inflammatory cytokine-related response (TGF-β and NF-κB) of tissues was observed in nano-ZnO exposed-mussels. Furthermore, the oxidative injury and apoptotic response could differentiate the effects of nano-ZnO from those of dissolved Zn in the mussels. This study revealed that oxidative stress and stress-related transcriptional responses to nano-ZnO were strongly modified by warming and season in the mussels. No single biomarker could be shown to consistently respond to nano-ZnO in all experimental groups, which implies that multiple biomarkers are needed to assess nano-ZnO toxicity to marine organisms under the variable environmental conditions of coastal habitats.

Stress response markers in the blood of São Tomé green sea turtles (Chelonia mydas) and their relation with accumulated metal levels

Metals are persistent worldwide being harmful for diverse organisms and having complex and combined effects with other contaminants in the environment. Sea turtles accumulate these contaminants being considered good bioindicator species for marine pollution. However, very little is known on how this is affecting these charismatic animals. São Tomé and Príncipe archipelago harbours important green sea turtle (Chelonia mydas) nesting and feeding grounds. The main goal of this study was to determine metal and metalloid accumulation in the blood of females C. mydas nesting in São Tomé Island, and evaluate the possible impacts of this contamination by addressing molecular stress responses. Gene expression analysis was performed in blood targeting genes involved in detoxification/sequestration and metal transport (mt, mtf and fer), and in antioxidant and oxidative stress responses (cat, sod, gr, tdx, txrd, selp and gclc). Micronuclei analysis in blood was also addressed as a biomarker of genotoxicity. Present results showed significant correlations between different gene expressions with the metals evaluated. The best GLM models and significant relationships were found for mt expression, for which 78% of the variability was attributed to metal levels (Al, Cu, Fe, Hg, Pb and Zn), followed by micronuclei count (65% - Cr, Cu, Fe, Hg, Mn and Zn), tdx expression (52% - Cd, Fe, Mn, Pb and Se), and cat expression (52% - As, Fe, Se and Cd x Hg). Overall, this study demonstrates that these green sea turtles are trying to adapt to the oxidative stress and damage produced by metals through the increased expression of antioxidants and other protectors, which raises concerns about the impacts on these endangered organisms' fitness. Furthermore, promising biomarker candidates associated to metal stress were identified in this species that may be used in future biomonitoring studies using C. mydas’ blood, allowing for a temporal follow-up of the organisms.

Sex Associated Effects of Noise Pollution in Stone Sculpin (Paracottus knerii) as a Model Object in the Context of Human-Induced Rapid Environmental Change.

Simple SummaryIn this comprehensive multidisciplinary study, we applied a novel multilevel approach to stone sculpins Paracottus knerii Dybowski, 1874, as model organisms and test for the first time the hypothesis of sex-dependent differences in response to long-term noise exposure in fish. The results testify that the stone sculpin females appeared to experience excessive stress, while the males showed adaptive recalibrations. These effects may be explained by a unique adaptive strategy of offspring care in the stone sculpin males and their biological role in reproductive behavior within the species. The findings obtained may help to elucidate the links between noise exposure in the context of human-induced rapid environmental change (HIREC), long-term sex-related changes in fishes, and the possible further evolutionary success of a species. Such HIREC modeling not only provides information about the potential consequences under anthropogenic pressure but also can help identify the natural mechanisms of stress resistance in different species, including those related to sex, and also contribute to the development of effective environmental management practices.This work simulates the consequences of HIREC using stone sculpins as model organisms. Sex-dependent effects of long-term noise exposure at mean sound pressure levels of 160–179 dB re 1 μPa (SPLpk–pk) were measured. We applied a multilevel approach to testing the stress response: a comparative analysis of the macula sacculi and an assessment of hematological and molecular stress responses. Noise exposure resulted in hair cell loss, changes in some cytometric parameters in blood, and an increase in the number of functionally active mitochondria in the red blood cells of males and its decrease in females, demonstrating a mitochondrial allostatic load and depletion of functional reserve. Finally, a statistically significant decrease in the telomerase activity of the auditory epithelium and a shortening of telomere length in the brain as molecular markers of stress were observed after noise exposure only in females. No significant decrease in telomerase activity and shortening of telomere length in nerve target tissues were observed in stressed males. However, we recorded an increase in the telomerase activity in male gonads. This sex-dependent difference in load may be associated with accelerated cellular aging in females and lower stress-related long-term risk in males. In this article, we discuss possible reasons for these noise-induced stress effects.

Transcriptomic response of the intertidal limpet Patella vulgata to temperature extremes

Global warming is challenging wild species in land and water. In the intertidal zone, species are already living at their thermal limits, being vulnerable even to small increases in maximum habitat temperatures. Knowledge of the mechanisms by which many intertidal zone species cope with elevated temperatures is limited. We analysed the molecular thermal stress response of the limpet Patella vulgata under slight and frequent (one-day), and extreme and rare (three-day) warming events. Using RNA-seq to assess differential gene expression among treatments, differing molecular responses were obtained in the two treatments, with more changes in gene expression after the three-day event; with one-third of the differentially expressed transcripts being down-regulated. However, across treatments we observed shifts in gene expression for common aspects of the heat stress response including intra-cellular communication, protein chaperoning, proteolysis and cell cycle arrest. Of the 71,675 transcripts obtained, only 259 were differentially expressed after both heating events. From these, 218 defined the core group (i.e. genes induced by thermal stress with similar expression patterns irrespective of the magnitude of the warming event). The core group was composed of already well-studied genes in heat stress responses in intertidal organisms (e.g. heat shock proteins), but also genes from less explored metabolic pathways, e.g. the ubiquitin system, which were also fundamental regardless of the magnitude of the imposed warming. Moreover, we have also identified 41 signaling genes (i.e. a set of genes responding to both events and with expression patterns specific to the intensity of thermal stress), principally including genes involved in the maintenance of extracellular structure that have previously not been identified as part of the response to thermal stress in intertidal zone organisms. These signaling genes will be useful heat stress molecular biomarkers for monitoring heat stress in natural populations.

Analysis of the cerebellar molecular stress response led to first evidence of a role for FKBP51 in brain FKBP52 expression in mice and humans

As the cerebellar molecular stress response is understudied, we assessed protein expression levels of hypothalamic-pituitary-adrenal (HPA) axis regulators and neurostructural markers in the cerebellum of a male PTSD mouse model and of unstressed vs. stressed male FK506 binding protein 51 (Fkbp5) knockout (KO) vs. wildtype mice. We explored the translatability of our findings in the Fkbp5 KO model to the situation in humans by correlating mRNA levels of candidates with those of FKBP5 in two whole transcriptome datasets of post-mortem human cerebellum and in blood of unstressed and stressed humans. Fkbp5 deletion rescued the stress-induced loss in hippocampal, prefrontal cortical, and, possibly, also cerebellar FKBP52 expression and modulated post-stress cerebellar expression levels of the glucocorticoid receptor (GR) and possibly (trend) also of glial fibrillary acidic protein (GFAP). Accordingly, expression levels of genes encoding for these three genes correlated with those of FKBP5 in human post-mortem cerebellum, while other neurostructural markers were not related to Fkbp5 either in mouse or human cerebellum. Also, gene expression levels of the two immunophilins correlated inversely in the blood of unstressed and stressed humans. We found transient changes in FKBP52 and persistent changes in GR and GFAP in the cerebellum of PTSD-like mice. Altogether, upon elucidating the cerebellar stress response we found first evidence for a novel facet of HPA axis regulation, i.e., the ability of FKBP51 to modulate the expression of its antagonist FKBP52 in the mouse and, speculatively, also in the human brain and blood and, moreover, detected long-term single stress-induced changes in expression of cerebellar HPA axis regulators and neurostructural markers of which some might contribute to the role of the cerebellum in fear extinction.

Responses of aquaculture fish to climate change‐induced extreme temperatures: A review

AbstractClimate change‐induced extreme temperature events are becoming more intense and frequent. For fish, temperature is the master abiotic factor that controls and limits fish development and physiology at all stages of aquaculture. Major physiological constraints at the individual level underpin changes in fish growth, hemato‐physiology, metabolism, immune and molecular stress responses to extreme temperature events. Extreme temperature impacts on fish vary among and within the diverse species depending on stress magnitude and associated factors. This review synthesizes the impacts of climate change‐induced extreme temperature events on neuroendocrine, oxidative, metabolic, osmotic, molecular, hemato‐biochemical, and immune responses in fish related to aquaculture. Besides, overall aspects of extreme temperature impacts on fish growth and reproduction are also discussed. Possible mitigation measures to maintain the fish's physiological fitness during extreme temperature events are also addressed. However, studies on the interactive effects of extreme temperature events with other associated environmental stressors are needed across a broader range of species to gain a better understanding of fish response and aquaculture performance during extreme temperature events. Moreover, for fish, studies on alternative species, development of stress‐tolerant strains, and the possibilities of nutritional mitigation measures as a potential option to combat extreme temperature stress are promising and should be pursued in future studies.

A proteomic analysis of the effect of ocean acidification on the haemocyte proteome of the South African abalone Haliotis midae

As a result of increasing CO2 emissions and the prevalence of global climate change, ocean acidification (OA) is becoming more pervasive, affecting many trophic levels, particularly those that rely on succinctly balanced ocean chemistry. This ultimately threatens community structures, as well as the future sustainability of the fishing/aquaculture industry. Understanding the molecular stress response of key organisms will aid in predicting their future survivability under changing environmental conditions. This study sought to elucidate the molecular stress response of the South African abalone, Haliotis midae, an understudied organism with high economic value, utilising a high throughput iTRAQ-based proteomics methodology. Adult abalone were exposed to control (pH 7.9) and experimental (pH 7.5) conditions for 12, 72 and 168 h, following which protein was isolated from sampled haemocytes and subsequently processed. iTRAQ-labelled peptides were analysed using mass spectrometry, while an array of bioinformatics tools was utilised for analysing the proteomic data. COG analysis identified “Cytoskeleton”, “Translation, ribosomal structure and biogenesis”, “Post-translational modification, protein turnover, chaperones”, and “Intracellular trafficking, secretion and vesicular transport” to be the most enriched functional classes, while statistical analysis identified a total of 33 up-regulated and 23 down-regulated effectors of OA stress in abalone. Several of the up-regulated proteins that were identified function in central metabolism (ENO1, PGK, DUOX1, GPD2), the stress/immune response (CAMKI, HSPA5/GRP78, MAPKI), and cytoskeleton, protein sorting and signal transduction (IQGAP1, MYO9B, TLN1, RDX, TCP-1/CCT, SNX6, CHMP1a, VPS13a). Protein-protein interactions were predicted using STRING DB, Cytoscape and Ingenuity Pathway Analysis, providing a model of the effects of OA on the H. midae haemocyte proteome. The data indicated that H. midae underwent a metabolic shift under OA conditions to utilize more energy-efficient mechanisms of ATP generation, while attempts at restoring haemocyte stabilisation and homeostasis were reflected by up-regulation of oxidative stress and cytoskeletal proteins. Our results support other molluscan studies that report a complex array of overlapping functions of both the stress and immune response systems. This interplay of the mounted stress and immune response is maintained and observed through the up-regulation of proteins involved in protein synthesis and turnover, as well as intracellular signalling and transport. The data presented in this study highlight the value of employing sensitive and robust -omics technologies for assessing the effects of changing environmental conditions on marine organisms.

Can dietary manipulation mitigate extreme warm stress in fish? The case of European seabass, Dicentrarchus labrax

Extreme climatic events affect all living organisms, including fish. This study evaluated growth, hematological, immune-antioxidant, and molecular stress responses to understand the acclimatization ability of European seabass, Dicentrarchus labrax fed on four different dietary supplements with subsequent exposure to an extreme ambient warm (32 oC) event. Fish were fed on diets supplemented with vitamins C (0.40%) and E (0.35%), propolis (0.45%), phycocyanin (0.30%), and β-glucan (0.30%) along with a control diet for 56 days, followed by 18 days of extreme warm exposure. Fish growth performance and survival were higher in fish fed on propolis followed by vitamins C & E and phycocyanin supplemented diets compared to fish fed on the control diet. During extreme warm exposure, red blood cells (RBC), hematocrit, hemoglobin, respiratory burst (RB), serum lysozyme activities (LSZ), and insulin-like growth factor 1 (Igf1) were significantly (P < 0.05) higher in fish fed on diets supplemented with propolis, vitamins C & E, and phycocyanin compared to fish fed on control diet. On day 18 of extreme warm stress, blood glucose and lactate levels were significantly higher in fish fed on control and β-glucan supplemented diets compared to the other three tested diets. On days 9 and 18, erythrocytic cellular abnormalities (ECA) and nuclear abnormalities (ENA) were significantly increased in fish fed on control and β-glucan diets (P < 0.05). Heat shock protein 70 (HSP70) and serum antioxidant enzymes [superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), and glutathione reductase (GR)] activities were comparatively lower in fish fed on propolis and vitamins C & E supplemented diets compared to the control diet on days 9 and 18 of extreme warm stress. Most of the repeatedly measured parameters indicated that European seabass fed on diets supplemented with propolis, vitamins C & E, and phycocyanin provide improved acclimation potential during extreme ambient warm (32°C) exposure.

Cellular Phosphorylation Signaling and Gene Expression in Drought Stress Responses: ABA-Dependent and ABA-Independent Regulatory Systems.

Drought is a severe and complex abiotic stress that negatively affects plant growth and crop yields. Numerous genes with various functions are induced in response to drought stress to acquire drought stress tolerance. The phytohormone abscisic acid (ABA) accumulates mainly in the leaves in response to drought stress and then activates subclass III SNF1-related protein kinases 2 (SnRK2s), which are key phosphoregulators of ABA signaling. ABA mediates a wide variety of gene expression processes through stress-responsive transcription factors, including ABA-RESPONSIVE ELEMENT BINDING PROTEINS (AREBs)/ABRE-BINDING FACTORS (ABFs) and several other transcription factors. Seed plants have another type of SnRK2s, ABA-unresponsive subclass I SnRK2s, that mediates the stability of gene expression through the mRNA decay pathway and plant growth under drought stress in an ABA-independent manner. Recent research has elucidated the upstream regulators of SnRK2s, RAF-like protein kinases, involved in early responses to drought stress. ABA-independent transcriptional regulatory systems and ABA-responsive regulation function in drought-responsive gene expression. DEHYDRATION RESPONSIVE ELEMENT (DRE) is an important cis-acting element in ABA-independent transcription, whereas ABA-RESPONSIVE ELEMENT (ABRE) cis-acting element functions in ABA-responsive transcription. In this review article, we summarize recent advances in research on cellular and molecular drought stress responses and focus on phosphorylation signaling and transcription networks in Arabidopsis and crops. We also highlight gene networks of transcriptional regulation through two major regulatory pathways, ABA-dependent and ABA-independent pathways, that ABA-responsive subclass III SnRK2s and ABA-unresponsive subclass I SnRK2s mediate, respectively. We also discuss crosstalk in these regulatory systems under drought stress.

SMURF2-mediated ubiquitin signaling plays an essential role in the regulation of PARP1 PARylating activity, molecular interactions, and functions in mammalian cells.

Poly(ADP-ribose) polymerase 1 (PARP1) is a key molecular stress sensor and response mediator implicated in multiple cellular functions in health and diseases. Despite its importance and intrinsic involvement in pivotal molecular and cellular processes, including DNA repair, transcription regulation, chromatin organization, and cell death, the regulatory mechanisms of PARP1 are poorly understood. In this study, we show that SMURF2, a HECT-type E3 ubiquitin ligase and suggested tumor suppressor, physically interacts with PARP1 in different cellular settings, directly ubiquitinates it in vitro and stimulates its PARylation activity in cells, the phenomenon that required SMURF2 E3 ubiquitin ligase function. Intriguingly, in the cellular environment SMURF2 was found to regulate the dynamic exchange of ubiquitin moieties on PARP1, mostly decreasing its monoubiquitination. Through the set of systematic mass spectrometry analyses conducted on SMURF2-modified cells, we identified on PARP1 18 lysine residues (out of 126 present in PARP1) as sites which ubiquitination was considerably affected by SMURF2. Subsequent site-directed mutagenesis coupled with in cellula ubiquitination and PARylation assays unveiled K222 as a critical site enabling a cross talk between SMURF2-modulated monoubiquitination of PARP1 and its activity, and pointed to K498, S507, and a KTR triad (K498/K521/K524) as the main auto-PARylation sites affected by SMURF2. The results also uncovered that SMURF2 controls PARP1 interactome, influencing its functions and expression in a context-dependent manner. Taken together, these findings suggest that SMURF2-mediated ubiquitin signaling plays an essential role in PARP1 regulation, beyond the regulation of its protein expression.