Stress Adaptation Mechanisms Research Articles

Recalcitrant cutaneous fungal infections-A growing problem.

The incidence and prevalence of recalcitrant cutaneous fungal infections is on the rise. Terbinafine-resistant Trichophyton has not only been widespread in India, but has also been reported in countries spread throughout the globe. Strains of yeasts such as Malassezia and Candida, which exist both as commensals and as pathogens to the human skin, have also been found to develop resistance to antifungals. Non-dermatophyte moulds which can colonize and infect damaged nails are especially difficult to treat, not only due to resistance, but also because of poor drug penetration of hard keratin. Psychosocial factors such as the indiscriminate broad-spectrum antifungal use in agriculture and in medicine, and poor adherence to hygienic measures to break the chain of infection contribute to the development of antifungal resistance. Such environments encourage fungi to develop various resistance mechanisms to withstand antifungal treatment. These include: (a) alteration of the drug target, (b) increasing efflux of drug/metabolites, (c) inactivation of drug, (d) bypass mechanisms or substitution of the pathway affected by the drug, (e) stress adaptation mechanisms and (f) biofilm formation. Understanding of such mechanisms and how they arise are crucial for development of new ways to prevent or overcome resistance. Novel antifungal treatments have recently been approved in the United States of America for treatment of vulvovaginal candidiasis. Ibrexafungerp (enfumafungin derivative) and oteseconazole (tetrazole) differ from their respective related drug classes of echinocandins and triazoles by having different structures, which lend these medicines advantage compared to traditional treatment by having a different binding site and more selectivity for fungi respectively. Other drugs designed to circumvent the known mechanisms of antifungal resistance are also at various phases of development. Concurrent measures at an institutional and individual level to address and limit inappropriate antifungal use to reduce development of antifungal resistance should be undertaken in a concerted effort to address this epidemic.

Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation

Systems-level assessments of protein-protein interaction (PPI) network dysfunctions are currently out-of-reach because approaches enabling proteome-wide identification, analysis, and modulation of context-specific PPI changes in native (unengineered) cells and tissues are lacking. Herein, we take advantage of chemical binders of maladaptive scaffolding structures termed epichaperomes and develop an epichaperome-based ‘omics platform, epichaperomics, to identify PPI alterations in disease. We provide multiple lines of evidence, at both biochemical and functional levels, demonstrating the importance of these probes to identify and study PPI network dysfunctions and provide mechanistically and therapeutically relevant proteome-wide insights. As proof-of-principle, we derive systems-level insight into PPI dysfunctions of cancer cells which enabled the discovery of a context-dependent mechanism by which cancer cells enhance the fitness of mitotic protein networks. Importantly, our systems levels analyses support the use of epichaperome chemical binders as therapeutic strategies aimed at normalizing PPI networks.

Ectopic over-expression of HaFT-1, a 14-3-3 protein from Haloxylon ammodendron, enhances acquired thermotolerance in transgenic Arabidopsis.

Haloxylon ammodendron, an important shrub utilized for afforestation in desert areas, can withstand harsh ecological conditions such as drought, high salt and extreme heat. A better understanding of the stress adaptation mechanisms of H. ammodendron is vital for ecological improvement in desert areas. In this study, the role of the H. ammodendron 14-3-3 protein HaFT-1 in thermotolerance was investigated. qRT-PCR analysis showed that heat stress (HS) priming (the first HS) enhanced the expression of HaFT-1 during the second HS and subsequent recovery phase. The subcellular localization of YFP-HaFT-1 fusion protein was mainly detected in cytoplasm. HaFT-1 overexpression increased the germination rate of transgenic Arabidopsis seeds, and the survival rate of HaFT-1 overexpression seedlings was higher than that of wild-type (WT) Arabidopsis after priming-and-triggering and non-primed control treatments. Cell death staining showed that HaFT-1 overexpression lines exhibited significantly reduced cell death during HS compared to WT. Transcriptome analysis showed that genes associated with energy generation, protein metabolism, proline metabolism, autophagy, chlorophyll metabolism and reactive oxygen species (ROS) scavenging were important to the thermotolerance of HS-primed HaFT-1 transgenic plants. Growth physiology analysis indicated that priming-and-triggering treatment of Arabidopsis seedlings overexpressing HaFT-1 increased proline content and strengthened ROS scavenging activity. These results demonstrated that overexpression of HaFT-1 increased not only HS priming but also tolerance to the second HS of transgenic Arabidopsis, suggesting that HaFT-1 is a positive regulator in acquired thermotolerance.

Valosin-containing protein (VCP/p97) is prognostically unfavorable in pediatric AML, and negatively correlates with unfolded protein response proteins IRE1 and GRP78: A report from the Children's Oncology Group.

The endoplasmic reticulum (ER) is the major site of protein synthesis and folding in the cell. ER-associated degradation (ERAD) and unfolded protein response (UPR) are the main mechanisms of ER-mediated cell stress adaptation. Targeting the cell stress response is a promising therapeutic approach in acute myeloid leukemia (AML). Protein expression levels of valosin-containing protein (VCP), a chief element of ERAD, were measured in peripheral blood samples from in 483 pediatric AML patients using reverse phase protein array methodology. Patients participated in the Children's Oncology Group AAML1031 phase 3 clinical trial that randomized patients to standard chemotherapy (cytarabine (Ara-C), daunorubicin, and etoposide [ADE]) versus ADE plus bortezomib (ADE+BTZ). Low-VCP expression was significantly associated with favorable 5-year overall survival (OS) rate compared to middle-high-VCP expression (81%versus 63%, p<0.001), independent of additional bortezomib treatment. Multivariable Cox regression analysis identified VCP as independent predictor of clinical outcome. UPR proteins IRE1 and GRP78 had significant negative correlation with VCP. Five-year OS in patients characterized by low-VCP, moderately high-IRE1 and high-GRP78 improved after treatment with ADE+BTZ versus ADE (66%versus 88%, p=0.026). Our findings suggest the potential of the protein VCP as biomarker in prognostication prediction in pediatric AML.

Insights into the Transcriptomics of Crop Wild Relatives to Unravel the Salinity Stress Adaptive Mechanisms.

The narrow genomic diversity of modern cultivars is a major bottleneck for enhancing the crop's salinity stress tolerance. The close relatives of modern cultivated plants, crop wild relatives (CWRs), can be a promising and sustainable resource to broaden the diversity of crops. Advances in transcriptomic technologies have revealed the untapped genetic diversity of CWRs that represents a practical gene pool for improving the plant's adaptability to salt stress. Thus, the present study emphasizes the transcriptomics of CWRs for salinity stress tolerance. In this review, the impacts of salt stress on the plant's physiological processes and development are overviewed, and the transcription factors (TFs) regulation of salinity stress tolerance is investigated. In addition to the molecular regulation, a brief discussion on the phytomorphological adaptation of plants under saline environments is provided. The study further highlights the availability and use of transcriptomic resources of CWR and their contribution to pangenome construction. Moreover, the utilization of CWRs' genetic resources in the molecular breeding of crops for salinity stress tolerance is explored. Several studies have shown that cytoplasmic components such as calcium and kinases, and ion transporter genes such as Salt Overly Sensitive 1 (SOS1) and High-affinity Potassium Transporters (HKTs) are involved in the signaling of salt stress, and in mediating the distribution of excess Na+ ions within the plant cells. Recent comparative analyses of transcriptomic profiling through RNA sequencing (RNA-Seq) between the crops and their wild relatives have unraveled several TFs, stress-responsive genes, and regulatory proteins for generating salinity stress tolerance. This review specifies that the use of CWRs transcriptomics in combination with modern breeding experimental approaches such as genomic editing, de novo domestication, and speed breeding can accelerate the CWRs utilization in the breeding programs for enhancing the crop's adaptability to saline conditions. The transcriptomic approaches optimize the crop genomes with the accumulation of favorable alleles that will be indispensable for designing salt-resilient crops.

Функціональні можливості серцево-судинної системи та стресостійкість осіб із різною теплочутливістю

Global warming affects people’s daily life, reduces endurance and work capacity. The purpose of our study is to investigate the functional capabilities of the circulatory system, adaptation potential and stress resistance of individuals with different heat sensitivity. To evaluate the activity of the cardiovascular system, an echocardiographic examination was performed. The reserve capabilities of the heart and the economy of its work were determined using the Robinson index and the Roufier test. Resistance to hypoxia was determined using the Stange and Genchi tests. The degree of oxygen assimilation was evaluated according to the values of maximum oxygen consumption. Cardiorespiratory fitness was determined based on the results of the Rufier test. Adaptation potential was established according to Baevsky’s method. Stress resistance was assessed by determining the response index of the cardiovascular system to psycho-emotional stress. It was established that in people with lower heat sensitivity, the activity of the cardiovascular system is more optimal and more economical and its reserve capacity is greater, compared to higher sensitivity individuals, in which the intensity of its work in an uneconomical mode is increased and the reserve potential is limited. Shorter breath retention time on inhalation and exhalation, respectively, in persons with higher sensitivity to heat indicates a worse oxygen supply to the body. Also, they have a poor ability to absorb oxygen and lower cardiorespiratory efficiency. The predominance of stress adaptation mechanisms and lower stress resistance were also revealed. Thus, our results show that individuals with a higher sensitivity to heat are particularly vulnerable to the negative effects of global warming, which should be taken into account for the prevention of cardiovascular and heat diseases in them in the future.

Transcriptome profiling, physiological, and biochemical analyses provide new insights towards drought stress response in sugar maple (Acer saccharum Marshall) saplings.

Sugar maple (Acer saccharum Marshall) is a temperate tree species in the northeastern parts of the United States and is economically important for its hardwood and syrup production. Sugar maple trees are highly vulnerable to changing climatic conditions, especially drought, so understanding the physiological, biochemical, and molecular responses is critical. The sugar maple saplings were subjected to drought stress for 7, 14, and 21 days and physiological data collected at 7, 14, and 21 days after stress (DAS) showed significantly reduced chlorophyll and Normalized Difference Vegetation Index with increasing drought stress time. The drought stress-induced biochemical changes revealed a higher accumulation of malondialdehyde, proline, and peroxidase activity in response to drought stress. Transcriptome analysis identified a total of 14,099 differentially expressed genes (DEGs); 328 were common among all stress periods. Among the DEGs, transcription factors (including NAC, HSF, ZFPs, GRFs, and ERF), chloroplast-related and stress-responsive genes such as peroxidases, membrane transporters, kinases, and protein detoxifiers were predominant. GO enrichment and KEGG pathway analysis revealed significantly enriched processes related to protein phosphorylation, transmembrane transport, nucleic acids, and metabolic, secondary metabolite biosynthesis pathways, circadian rhythm-plant, and carotenoid biosynthesis in response to drought stress. Time-series transcriptomic analysis revealed changes in gene regulation patterns in eight different clusters, and pathway analysis by individual clusters revealed a hub of stress-responsive pathways. In addition, qRT-PCR validation of selected DEGs revealed that the expression patterns were consistent with transcriptome analysis. The results from this study provide insights into the dynamics of physiological, biochemical, and gene responses to progressive drought stress and reveal the important stress-adaptive mechanisms of sugar maple saplings in response to drought stress.

Genome-Wide Identification and Characterization of Olfactory Receptor Genes in Silver Sillago (Sillago sihama).

Olfactory receptor (OR) genes are essential in the specific recognition of diverse stimuli in fish. In this study, a total of 141 OR genes were identified in silver sillago (Sillago sihama), a marine fish sensitive to environmental stimuli, including 112 intact genes, 26 truncated genes, and three pseudogenes. A phylogenetic tree analysis elucidated that the OR genes of S. sihama were classified into six groups, of which β, γ, δ, ε, and ζ groups belonged to type I, and the η group belonged to type II. The type I OR genes contained almost all conserved motifs (n = 62), while type II OR genes mainly retained conserved motifs 7(3), 1, 10, 4, and 2 (n = 39). OR genes were mainly distributed on LG1, LG9, LG11, and LG12. Of all OR genes, 36.23% (50 genes) showed significant expansion in S. sihama. Ka/Ks analysis demonstrated that 227 sites were under purifying selection, while 12 sites were under positive selection, including eight genes in the OR2A12 gene subfamily. Sixty-one genes (44.20%) displayed differential expression under hypoxic stress. The identified OR genes explored the mechanism of environmental stress and ecological adaptation of S. sihama, and provided valuable genomic resources for further research on the olfaction of teleosts.

Seed color represents salt resistance of alfalfa seeds (Medicago sativa L.): Based on the analysis of germination characteristics, seedling growth and seed traits.

Alfalfa (Medicago sativa L.) has significant feed value and ecological improvement function of marginal land. The difference in the maturity period of seeds in the same lots may be a mechanism of environmental adaptation. Seed color is a morphological trait associated with seed maturity. A clear understanding of the relationship between the seed color and seed stress resistance is helpful for seed selection for marginal land. This study evaluated the seed germination parameters (germinability and final germination percentage) and subsequent seedling growth (sprout height, root length, fresh and dry weight) of alfalfa under different salt stress, measured the electrical conductivity, water uptake, seed coat thickness, and endogenous hormone content in alfalfa seeds with different colors (green, yellow and brown). The results showed that seed color significantly influenced the seed germination and seedling growth. The germination parameters and seedling performance of brown seeds were significantly lower than that of green and yellow seeds under different salt stress. The germination parameters and seedling growth of brown seed declined most obviously with the aggravation of salt stress. The results suggested that brown seeds were less resistant to salt stress. Seed color had a significant effect on electrical conductivity, and it indicated that the yellow seeds had higher vigor. The seed coat thickness of various colors did not differ significantly. The seed water uptake rate and hormone content (IAA, GA3, ABA) in brown seeds were higher than that in green and yellow seeds, while the (IAA+GA3)/ ABA in yellow seeds were higher than green and brown seeds. The alterations in seed germination and seedling performance among seed colors are likely due to the combination effect of the content and balance between IAA+GA3 and ABA. These results could improve the understanding of stress adaptation mechanisms of alfalfa and provide a theoretical basis for screening alfalfa seeds with high stress resistance.

Vitamin D regulates ion regulation by affecting the ionocyte differentiation in zebrafish (Danio rerio) larvae

Freshwater teleosts frequently face the stress of varied ion and pH levels; therefore, they have developed related defense mechanisms to maintain the homeostasis of body-fluid ion and acid-base balance. The different subtypes of ionocytes expressed in the branchial epithelium of adult fish or the skin of larvae are the major sites for fish ion regulation. 1α,25-dihydroxyvitamin D3 (1α,25(OH)2D3), the bioactive form of vitamin D, is a steroid hormone that is involved in the regulation of Ca2+ uptake and acid secretion in teleosts. Our results revealed that 1α,25(OH)2D3 levels were not changed in zebrafish larvae upon exposure to low Na+ freshwater compared to normal freshwater. In contrast, 1α,25(OH)2D3 levels were substantially higher in fish exposed to acidic and low Ca2+ freshwater than in those exposed to normal freshwater. Some hormones regulate ion regulation and acid secretion by modulating ionocyte differentiation and/or proliferation in teleosts; however, the role of vitamin D in this process is unclear. Zebrafish larvae were used as a model in the present study to explore the effect of vitamin D on ionocyte proliferation and/or differentiation. The present study indicated that 1α,25(OH)2D3 treatment increased the number of foxi3a-positive cells, ionocyte progenitors, and mature ionocytes. However, the number of P63-positive epidermal stem cells did not change in the zebrafish larvae treated with 1α,25(OH)2D3. These results revealed that vitamin D exerts a positive effect on the number of ionocytes by increasing the differentiation of ionocytes. Increased ionocyte differentiation by vitamin D is suggested to elevate the capacity of ion regulation and acid secretion in zebrafish to cope with external stress. The present findings indicate the role of vitamin D in the regulation of ionocyte differentiation and provide new insights into the mechanisms of stress adaptation of fish.

Transcriptome Profiling of Gossypium anomalum Seedlings Reveals Key Regulators and Metabolic Pathways in Response to Drought Stress.

Drought stress is a key limiting factor for cotton (Gossypium spp.) growth, production, development, and production worldwide. Some wild diploid cotton species are remarkably tolerant of water deficit and constitute an important reservoir for understanding the molecular mechanisms of Gossypium spp. drought tolerance and improving cultivated upland cotton. Here, we utilized RNA-Seq technology to characterize the leaf transcriptomes of a wild African diploid cotton species, Gossypium anomalum, under drought stress. A total of 12,322 differentially expressed genes (DEGs) were identified after mapping valid clean reads to the reference genome of G. anomalum, of which 1243 were commonly differentially expressed at all stages of drought stress. These genes were significantly enriched for molecular functions Gene Ontology terms related to cytoskeleton, hydrolase activity, cellular redox, and binding. Additionally, a substantial proportion of enriched biological process terms concerned cell or subcellular processes, while most in the cellular components category concerned membrane function and photosynthesis. An enrichment analysis against the Kyoto Encyclopedia of Genes and Genomes showed the top significantly enriched pathways to be photosynthesis-antenna proteins, amino sugar and nucleotide sugar metabolism, starch and sucrose metabolism, MAPK signaling pathway, glutathione metabolism, and plant hormone signal transduction. The DEGs also exhibited interestingly significant enrichments for drought stress-induced tandemly repeated genes involved in iron ion binding, oxidoreductase activity, heme binding, and other biological processes. A large number of genes encoding transcription factors, such as MYB, bHLH, ERF, NAC, WRKY, and bZIP, were identified as playing key roles in acclimatizing to drought stress. These results will provide deeper insights into the molecular mechanisms of drought stress adaptation in Gossypium spp.

Biochemical and metabolic signatures are fundamental to drought adaptation in PGPR Enterobacter bugandensis WRS7.

Drought alone causes more annual loss in crop yield than the sum of all other environmental stresses. There is growing interest in harnessing the potential of stress-resilient PGPR in conferring plant resistance and enhancing crop productivity in drought-affected agroecosystems. A detailed understanding of the complex physiological and biochemical responses will open up the avenues to stress adaptation mechanisms of PGPR communities under drought. It will pave the way for rhizosphere engineering through metabolically engineered PGPR. Therefore, to reveal the physiological and metabolic networks in response to drought-mediated osmotic stress, we performed biochemical analyses and applied untargeted metabolomics to investigate the stress adaptation mechanisms of a PGPR Enterobacter bugendensis WRS7 (Eb WRS7). Drought caused oxidative stress and resulted in slower growth rates in Eb WRS7. However, Eb WRS7 could tolerate drought stress and did not show changes in cell morphology under stress conditions. Overproduction of ROS caused lipid peroxidation (increment in MDA) and eventually activated antioxidant systems and cell signalling cascades, which led to the accumulation of ions (Na+, K+, and Ca2+), osmolytes (proline, exopolysaccharides, betaine, and trehalose), and modulated lipid dynamics of the plasma membranes for osmosensing and osmoregulation, suggesting an osmotic stress adaption mechanism in PGPR Eb WRS7. Finally, GC-MS-based metabolite profiling and deregulated metabolic responses highlighted the role of osmolytes, ions, and intracellular metabolites in regulating Eb WRS7 metabolism. Our results suggest that understanding the role of metabolites and metabolic pathways can be exploited for future metabolic engineering of PGPR and developing bio inoculants for plant growth promotion under drought-affected agroecosystems.

Алармоны как фактор персистенции бактерий

The treatment of bacterial infections with antibiotics is significantly complicated because of the adaptive mechanisms employed by bacteria. Chronic and recurrent infections are often linked to bacterial persis-tence, biofilm formation, and antibiotic tolerance. These processes result in reduced metabolic activity, rendering bacteria insensitive to conventional antibiotics that primarily target actively growing cells. The stringent re-sponse, regulated by (p)ppGpp alarmone molecules, serves as a stress adaptation mechanism. It is conserved across numerous bacterial species and plays an important role in long-term survival under nutrient-depleted con-ditions. (p)ppGpp alarmones also play a significant role in bacterial persistence and the formation of biofilms. The pursuit of novel antibacterial agents that specifically target (p)ppGpp synthesis, thereby inhibiting the strin-gent response, presents a promising strategy in the battle against bacterial infections. In this context, alarmone synthetase inhibitors emerge as promising candidates for clinical application, as they have demonstrated their effectiveness in suppressing bacterial survival mechanisms, inhibiting biofilm formation, and reducing antibiotic tolerance and bacterial persistence.

Role of epigenetics and the high-throughput sensing techniques to detect stress adaptation mechanisms in crop plants: A mini review

Deciphering the epigenetic code, which will pave the road to understanding the link between genotypic and phenotypic variety, is one of the most fascinating topics of modern genetics. This may significantly contribute to our understanding of the complex mechanisms underlying plants\&amp;apos; epigenomic responses to environmental stressors in the context of climate change. As a result of epigenetic regulatory processes such DNA methylation, histone structural modification, and RNA-based regulatory mechanisms, it is hypothesised that chromatin will alter. The interactions of regulatory proteins, including various types of transcription factors, with DNA and chromatin would result in a variety of structural alterations that would impact how genes are expressed. High throughput sequencing techniques have recently been developed to better understand epigenomic alterations in the genome. The use of pharmacological and genetic methods to modify these networks should be made simpler by these strategies. The cytosine methylation of a cell\&amp;apos;s genomic regions or active cistrons, in particular, is an epigenetic change that these approaches are successful at identifying and studying. To speed up breeding programs for crop improvement, numerous investigations on the interactions between genotype, phenotype, and epigenotype utilising epiGWAS are being carried out. This review will briefly explore the role of epigenetic approaches and high throughput sensing techniques in order to better understand plant stress responses. The value of epigenomic methods and strategies in the nascent \&amp;quot;omic\&amp;quot; sciences is being emphasized

How to survive pig farming: Mechanism of SCCmec element deletion and metabolic stress adaptation in livestock-associated MRSA.

Previous research on methicillin susceptible Staphylococcus aureus (MSSA) belonging to livestock-associated (LA-) sequence type (ST) 398, isolated from pigs and their local surroundings, indicated that differences between these MSSA and their methicillin resistant predecessors (MRSA) are often limited to the absence of the staphylococcal cassette chromosome mec (SCCmec) and few single nucleotide polymorphisms. So far, our understanding on how LA-MRSA endure the environmental conditions associated with pig-farming as well as the putative impact of this particular environment on the mobilisation of SCCmec elements is limited. Thus, we performed in-depth genomic and transcriptomic analyses using the LA-MRSA ST398 strain IMT38951 and its methicillin susceptible descendant. We identified a mosaic-structured SCCmec region including a putative replicative SCCmecVc which is absent from the MSSA chromosome through homologous recombination. Based on our data, such events occur between short repetitive sequences identified within and adjacent to two distinct alleles of the large cassette recombinase genes C (ccrC). We further evaluated the global transcriptomic response of MRSA ST398 to particular pig-farm associated conditions, i.e., contact with host proteins (porcine serum) and a high ammonia concentration. Differential expression of global regulators involved in stress response control were identified, i.e., ammonia-induced alternative sigma factor B-depending activation of genes for the alkaline shock protein 23, the heat shock response and the accessory gene regulator (agr)-controlled transcription of virulence factors. Exposure to serum transiently induced the transcription of distinct virulence factor encoding genes. Transcription of genes reported for mediating the loss of methicillin resistance, especially ccrC, was not significantly different compared to the unchallenged controls. We concluded that, from an evolutionary perspective, bacteria may save energy by incidentally dismissing a fully replicative SCCmec element in contrast to the induction of ccr genes on a population scale. Since the genomic SCCmec integration site is a hot-spot of recombination, occasional losses of elements of 16 kb size may restore capacities for the uptake of foreign genetic material. Subsequent spread of resistance, on the other hand, might depend on the autonomous replication machinery of the deleted SCCmec elements that probably enhance chances for reintegration of SCCmec into susceptible genomes by mere multiplication.

Abstract B079: Unfolding the role of cell state on stress granule heterogeneity and function in pancreatic cancer

Abstract Phenotypic and functional heterogeneity is a common feature of cancer cells that poses as a major therapeutic challenge. Previous work has implicated the phase-separation organelles, stress granules (SGs), as important mediators of tumorigenesis that function as a mutant KRAS-driven stress-adaptive mechanism to enhance cellular fitness. We have found that mutant KRAS pancreatic cancer cells show a high degree of intra-cellular heterogeneity in SG formation, ranging from no SGs to markedly high levels. Given the function of SGs in cancer cell fitness and the role of cancer cell heterogeneity in tumor progression and therapeutic response, we sought to understand the determinants of SG heterogeneity and its role in KRAS-driven tumorigenesis. Here, we show that the capability of KRAS mutant pancreatic cancer cells to form SGs is dependent on the cell cycle state, with cells in G2 phase showing markedly enhanced SG formation. We demonstrate that this heterogeneity in SG formation is determined by a cell cycle specific interplay between the activator of cell death, Caspase 3, and the SG regulator molecule 15-deoxy-delta-12,14-prostaglandin J2 (15-d-PGJ2). We show that 15-d-PGJ2 production in G2 is driven by calcium-dependent phospholipase A2 (cPLA2) activity and is subject to regulation by Caspase 3. Our results show that the capacity of Caspase 3 to cleave and inactivate the cPLA2 protein is specifically suppressed in G2, thus leading to upregulation of cPLA2 activity and 15-d-PGJ2 levels. As such, we find that 15d-PGJ2 levels are highest in G2 and that the inhibition of cPLA2 blocks SG formation in G2 specifically. Accordingly, exogenous 15d-PGJ2 rescues SG formation in cPLA2-inhibited G2 cells. In addition, we demonstrate that Caspase 3 inhibition increases 15-d-PGJ2 levels and stimulates SG formation in asynchronous cells, mirroring the natural suppression of Caspase 3 seen in G2. Importantly, we show that heterogeneity in SG levels translates into a functional heterogeneity whereby SG inhibition preferentially sensitizes cells in G2 to stress stimuli. Altogether these data support a model whereby SG formation integrates cues from cell death and proliferative signals and indicate that SG levels may be critical to achieve a clear signal for cell cycle progression at the G2/M border. Furthermore, our findings suggest that cancer cells may show differential sensitivity to anti-SG therapy as they progress through the cell cycle, and pairing such therapy with G2-synchronizing chemotherapeutics may increase susceptibility and thus treatment efficacy. Citation Format: Alexandra Redding, Guillaume Fonteneau, Elda Grabocka. Unfolding the role of cell state on stress granule heterogeneity and function in pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer; 2022 Sep 13-16; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2022;82(22 Suppl):Abstract nr B079.

Differential expression and regulation of HSP70 gene during growth phase in ruminants in response to heat stress

Heat shock proteins regulate the physiological mechanism of heat stress adaptation at cellular level. The present investigation was carried out to analyse the HSP70 gene regulation in various growth stage in ruminants in peripheral blood mononuclear cells (PBMCs). The relationship between HSP gene expression and thermotolerance in age-specific manner in ruminants has not been analysed. Therefore m-RNA HSP70 expression level was examined in different age groups of Jamunpari goat during hot climatic conditions. The experiment was carried out in 32 animals of Jamunapari goat belonging to the age groups of 3-months, 9-months, 12-months, and adults (2–3 year). Total RNA was isolated from peripheral blood mononuclear cells. The physiological response such as rectal temperature (RT), respiration rate (RR) and heart rate (HR) was used as indicator to heat stress. Temperature Humidity Index (THI) was used as an indicator of severity of environmental stress. The THI range varied from 82.00–92.08 during experimental period. The m-RNA HSP70 expression level at 9-month age of animals was up-regulated and significantly higher than other age groups. It was observed that the level of HSP70 transcripts in PBMCs was highest at 9-month age group, and age-related decline in HSP70 expression was observed in adult age. Based on the physiological response, the contrasting heat-stress phenotypes were recognised as heat stress susceptible (HSS) and heat stress tolerant (HST) individuals and the expression of m-RNA HSP70 was analysed at different ages in response to chronic heat stress. The differential mRNA expression of HSS individuals at 3 and 9-month of age showed the highest fold expression than HST. Age and phenotype had significant effect (p < 0.01) on the crossing point (CP) value. The m-RNA HSP70 gene expression in different age groups was correlated with heat stress tolerance and this could be used as biomarker for breeders to analyse the HSP response in -vivo in ruminants.

Reference Gene Selection for qPCR Analysis in Schima superba under Abiotic Stress.

Quantitative real-time PCR (qPCR) is an indispensable technique for gene expression analysis in modern molecular biology. The selection and evaluation of suitable reference genes is a prerequisite for accurate gene expression analysis. Schima superba is a valuable tree species that is environmentally adaptable and highly fire-resistant. In this study, 12 candidate reference genes were selected to check their stability of gene expression in different tissues under abiotic stresses: cold stress, salt stress, and drought stress by ΔCt, geNorm, NormFinder, BestKeeper, and RefFinder. The results indicated that AP-2 was the most stably expressed overall and for the cold stress and drought stress. eIF-5α gene expression was the most stable under the salt stress treatment, while UBQ expression was the most stable across mature leaves, shoots, stems, and roots. In contrast, UBC20, GAPDH, and TUB were the least stably expressed genes tested. This study delivers valid reference genes in S. superba under the different experimental conditions, providing an important resource for the subsequent elucidation of the abiotic stress adaptation mechanisms and genes with biological importance.

Evaluation of the Circadian Rhythm Neuropeptides and Hippocampus in Rat Model of Posttraumatic Stress

ABSTRACT We investigated hippocampus weight and the levels of circadian rhythm-related neuropeptides; Somatostatin (SST), Vasoactive Intestinal Peptide (VIP), Neuropeptide Y (NPY), Melatonin (MLT), and Corticotrophin Releasing Hormone (CRH) in plasma (p) and hippocampus tissue (h) of rats with posttraumatic stress (PTS) symptoms. Thirty four healthy Spraque-Dawley rats were randomly divided into control and PTS groups. Single prolonged stress (SPS) model was used to mimic PTS. Elevated Plus Maze Test (EPMT) was used to assess anxiogenic behavior. Rats were sacrificed under anesthesia and hippocampus weight, SST, NPY, CRH, MLT, and VIP levels were measured by the Enzyme-Linked-Immuno-Sorbent-Assay method. Hippocampus weights in the PTS group were significantly lower compared to controls. NPYh and MLTh levels were significantly higher in the PTS group. NPYh and MLTh and MLTp were significantly higher in the PTS. CRH, VIP, and SST levels in hippocampus tissue and CRH, NPY, VIP, and SST in plasma did not differ between groups. A positive correlation was found between MLTh and VIPh tissue levels. There was a positive correlation between NPYh and VIPh levels, and a high degree of positive correlation between NPYp and VIPp levels. As far as we know, this is the first study in which 5 neuropeptides’ hippocampus tissue and plasma levels were evaluated together in a PTSD model. Hippocampus weights may tend to decrease in PTS. Especially, NPY and MLT from circadian rhythm neuropeptides play an important role in PTS etiology. After the 7th day of trauma exposure, the hippocampal NPY and MLT levels may increase as a stress-adaptive mechanism in PTS.

Metabolomic and Transcriptomic Analyses of Lycium barbarum L. under Heat Stress

Heat stress has a strong and detrimental effect on plant growth and yield. Goji berry or wolfberry (Lycium barbarum L.) is a dual-purpose medicinal and food plant but an increase in high temperatures has caused a serious decline in wolfberry yield and quality. In this study, we first explored the heat stress responses of Goji berry, and found that heat stress adaptation mechanisms fluctuated over 48 h. Moreover, L.barbarum 1402 was more heat resistant while L.barbarum Ningqi No. 7 (N7) was sensitive to high temperatures, in which amino acids and alkaloids played key roles; expression and accumulation timing was also crucial. That is, 1402 responded to heat stress rapidly starting at 1 h under high temperature, activated related genes, and accumulated metabolites earlier in the amino acid metabolic pathway compared to N7, which responded to heat stress starting at 3 h under high temperature. Thus, 1402 resisted high temperatures much earlier and better compared to N7. Furthermore, joint transcriptome and metabolome analysis results showed that L-phenylalanine, L-tyrosine, N-benzylformamide, N-benzylmethylene isomethylamine, lysoPC 19:1, and N-acetyl-D-glucosamine-1-phosthate, as well as their related genes, were higher in content, or earlier in expression, in 1402 compared to N7 under heat treatment. This study initially elucidates that Goji berry 1402 has a better tolerance to heat stress than N7 for earlier and higher expression or accumulation of amino acids and alkaloids when related to high temperatures.