Alkaline Stress Research Articles

IDENTIFICATION, SEPARATION, AND CHARACTERIZATION OF DEGRADATION PRODUCTS OF TRIAMCINOLONE HEXACETONIDE USING LC AND LC-MS/MS

Objective: The study aimed to separate the forced degradation products of Triamcinolone hexacetonide using HPLC and characterize the degradation product by LC-MS/MS fragmentation pattern. Methods: Triamcinolone hexacetonide (THA) and its primary degradation products were identified using a liquid chromatography-mass spectrometry/Mass spectrometry (LC-MS/MS) approach. The degradation study was based on in-depth stress testing with acid, base, peroxide, heat, and light. A Zorbax SB C18 column and a greener mobile phase composed of methanol and 10 mmol ammonium acetate buffer in water at pH 3 were employed to accomplish separation and quantitation at a flow rate of 0.7 ml/min in an isocratic mode with a 239 nm detection wavelength. Results: A major degradation product of the drug was obtained in acidic and alkaline stress conditions. The drug was found to be stable for all other stress conditions. The LC-MS/MS analysis results of the active pharmaceutical ingredient and resulting product after degradation were interpreted to identify the novel degradation product and fragments. The developed method was validated as per International Council for Harmonization (ICH) guidelines. The square root of the correlation coefficients, which indicated linearity for THA in 50 to 150 % of the workload, was 0.99. Method Precision assay was performed on six different preparations, percentage relative standard deviation (% RSD) of assay value is 0.17 % and system precision is 0.30 %. In accuracy, overall % RSD of 50 %, 100 %, and 150 % in triplicate is 0.95. Conclusion: It is concluded that the drug is stable to all other stress conditions except for acidic and alkaline stress conditions and generates a novel degradation product. The developed LC (Liquid chromatography) method separates and identifies the degradation product.

Effects of saline and alkaline stresses on the survival, growth, and physiological responses in juvenile mandarin fish (Siniperca chuatsi)

Saline-alkaline water is widely distributed and abundant worldwide. It plays a critical role in the advancement of the aquaculture and the comprehensive utilization of saline-alkaline resources. The mandarin fish (Siniperca chuatsi), a freshwater species with high economic value, and has potential for cultivation in such environments. We evaluated its tolerance to acute salinity and alkalinity, and investigated the growth and physiological changes under chronic saline and alkaline stress. Initially, the effects of acute salinity and alkalinity were estimated on mandarin fish (4.33 ± 0.31 g), and found that the LC50 for 96 h of salinity and alkalinity was 13.91 ppt and 13.45 mmol/L, respectively. Subsequently, over a 60-day period, we measured the survival, growth performance, and physiological responses in mandarin fish (22.53 ± 0.60 g) reared in freshwater (FW, the control group), saline water (SW; fixed salinity of 5 ppt) and alkaline water (AW; carbonate alkalinity of 8 mmol/L). The results indicated that neither SW nor AW group adversely affected survival rates, and growth performance in SW group was significantly superior to the FW group. Additionally, compared to the FW group， the AW group had a lower hepatosomatic index, while the SW group exhibited significantly higher meat content. In terms of physiological responses, the AW group showed elevated V-H+-ATPase (VHA) activity, whereas the SW group had decreased prolactin (PRL) levels. Furthermore, the mitochondria-rich cells (MRCs) in mandarin fish underwent morphological adaptations to saline and alkaline stress, indicating their role in osmoregulation and acid-base balance. Our results suggested that the mandarin fish demonstrated good tolerance to both salinity and alkalinity, indicating suitability for breeding in low saline and alkaline water. This research contributes to a broader understanding of saline and alkaline tolerance in aquaculture species and provides theoretical support for the cultivation of mandarin fish in saline-alkaline water. Moreover, the results of this study suggested that saline-alkaline areas and agricultural irrigation water in the world could be successfully utilized for sustainable aquaculture.

Adaptation responses of different ecotypes of Leymus chinensis to saline–alkaline stress

IntroductionSoil salinity–alkalinity has emerged as a global problem affecting many ecosystems, including grassland. Plants evolve into different ecotypes to adapt to various environments. Leymus chinensis widely distributed in the eastern Eurasian steppe, has evolved into two main ecotypes: yellow–green(YG) and gray–green (GG). Studies on the adaption mechanisms of both ecotypes in response to saline–alkaline stress are limited.MethodsIn this study, the growth and physiological traits of ecotypes YG and GG in soils with different salinity–alkalinity levels—severe saline–alkaline soil (SS), moderate saline–alkaline soil (MS), and light saline–alkaline soil (LS)—were studied. After exposure to saline–alkaline stress for 15 months, the L. chinensis ecotypes exhibited significant differences in the growth characteristics.ResultsThe specific leaf area, individual tiller biomass, net photosynthetic rate, and potassium content of the two ecotypes under MS conditions were significantly higher than or similar to those under LS conditions. This indicates that L. chinensis showed a certain degree of tolerance to saline–alkaline environments under MS conditions. Saline–alkaline stress increased the tillers by 56% in GG and reduced them by 26% in YG, and did not alter the SLA and the number of individual tiller leaves of GG but reduced that of YG. Moreover, with increasing levels of saline–alkaline stress, ecotype GG exhibited an increase in net photosynthetic rate (Pn), while ecotype YG showed insignificant changes. Under SS, GG exhibited higher Pn and chlorophyll content than YG. Additionally, with the increase in the saline–alkaline stress level, the Na+ content increased, but GG exhibited a significantly lower Na+ content than YG. Conversely, the K+ and Ca2+ contents and the K+/Na+ and Ca2+/Na+ ratios decreased, but GG exhibited higher values than YG.DiscussionGG reduced Na+ by absorption by increasing tillers. Additionally, GG absorbed more K+ and Ca2+ ions, thereby maintaining higher K+/Na+ and Ca2+/Na+ ratios than YG under saline–alkaline stress. These adaptive strategies enable ecotype GG to grow and maintain normal physiological functions under high saline–alkaline stress. The findings of this study hold practical significance for enhancing the economic and ecological value of saline–alkaline grasslands.

Exogenous melatonin delays leaves senescence and enhances saline and alkaline stress tolerance in grape seedlings

ABSTRACT Saline and alkaline stress is one of the major abiotic stresses facing agricultural production, which severely inhibits the growth and yield of plant. The application of plant growth regulators can effectively prevent crop yield reduction caused by saline and alkaline stress. Exogenous melatonin (MT) can act as a signaling molecule involved in the regulation of a variety of physiological processes in plants, has been found to play a key role in enhancing the improvement of plant tolerance to abiotic stresses. However, the effects of exogenous MT on saline and alkaline tolerance of table grape seedlings and its mechanism have not been clarified. The aim of this study was to investigate the role of exogenous MT on morphological and physiological growth of table grape seedlings ( Vitis vinifera L.) under saline and alkaline stress. The results showed that saline and alkaline stress resulted in yellowing and wilting of grape leaves and a decrease in chlorophyll content, whereas the application of exogenous MT alleviated the degradation of chlorophyll in grape seedling leaves caused by saline and alkaline stress and promoted the accumulation of soluble sugars and proline content. In addition, exogenous MT increased the activity of antioxidant enzymes, which resulted in the scavenging of reactive oxygen species (ROS) generated by saline and alkaline stress. In conclusion, exogenous MT was involved in the tolerance of grape seedlings to saline and alkaline stress, and enhanced the saline and alkaline resistance of grape seedlings to promote the growth and development of the grape industry in saline and alkaline areas.

Improving strawberry plant resilience to salinity and alkalinity through the use of diverse spectra of supplemental lighting

BackgroundThis study explores the impact of various light spectra on the photosynthetic performance of strawberry plants subjected to salinity, alkalinity, and combined salinity/alkalinity stress. We employed supplemental lighting through Light-emitting Diodes (LEDs) with specific wavelengths: monochromatic blue (460 nm), monochromatic red (660 nm), dichromatic blue/red (1:3 ratio), and white/yellow (400–700 nm), all at an intensity of 200 µmol m-2 S-1. Additionally, a control group (ambient light) without LED treatment was included in the study. The tested experimental variants were: optimal growth conditions (control), alkalinity (40 mM NaHCO3), salinity (80 mM NaCl), and a combination of salinity/alkalinity.ResultsThe results revealed a notable decrease in photosynthetic efficiency under both salinity and alkalinity stresses, especially when these stresses were combined, in comparison to the no-stress condition. However, the application of supplemental lighting, particularly with the red and blue/red spectra, mitigated the adverse effects of stress. The imposed stress conditions had a detrimental impact on both gas exchange parameters and photosynthetic efficiency of the plants. In contrast, treatments involving blue, red, and blue/red light exhibited a beneficial effect on photosynthetic efficiency compared to other lighting conditions. Further analysis of JIP-test parameters confirmed that these specific light treatments significantly ameliorated the stress impacts.ConclusionsIn summary, the utilization of blue, red, and blue/red light spectra has the potential to enhance plant resilience in the face of salinity and alkalinity stresses. This discovery presents a promising strategy for cultivating plants in anticipation of future challenging environmental conditions.

Untargeted LC–MS metabolomics reveals the metabolic responses in the Eriocheir sinensis gills exposed to salinity and alkalinity stress

In recent years, saline–alkaline aquaculture development has become an important measure for China to expand its fishery development space to ensure food safety. Previous studies have verified that salinity and alkalinity positively influence the quality of Chinese mitten crabs (Eriocheir sinensis). However, the regulatory mechanism of E. sinensis endures saline–alkaline stress which remains obscure. This study investigated the metabolic changes in puberty-molting E. sinensis gills exposed to freshwater (FW), sodium chloride salinity of 5 ppt (SW), and carbonate alkalinity 10.00 mmol/L (AW) for 50 days using untargeted liquid chromatography-mass spectrometry metabolomics (LC–MS). A total of 5802 (positive-ion mode) and 6520 (negative-ion mode) peaks were extracted by LC–MS, respectively. A total of 188 (50 upregulated and 138 downregulated), 141 (94 upregulated and 47 downregulated), and 130 (87 upregulated and 43 downregulated) significantly regulated metabolites (SRMs) were observed in the FW–SW, FW–AW, and SW–AW treatments, respectively, wherein 42 generic SRMs were also found by Venn diagram analysis. Seven of the top 10 SRMs with the highest (variable importance in projection) VIP values were similarly identified in FW–SW and SW–AW. Integrated analysis of key metabolic pathways revealed glycerophospholipid, choline in cancer, phenylalanine, and butanoate metabolism. Overall, significant differences were observed in the metabolites and key metabolic pathways of E. sinensis gill exposed to salinity and alkalinity stress. These results will be helpful in understanding the environmental adaptability of aquatic crustaceans to saline–alkaline water.

Effects of Bacillus subtilis on Rose Growth Promotion and Rhizosphere Microbial Community Changes under Saline–Alkaline Stress

This study investigated the impact of Bacillus subtilis on plant growth and the rhizosphere microbial community in rose cultivation under saline–alkaline stress. Saline–alkaline stress was simulated with varying salt and alkali levels. Bacillus subtilis was introduced, and 16S rRNA high-throughput sequencing was conducted to analyze the root microbial community. Introduction of Bacillus subtilis significantly promoted rose growth and mitigated saline–alkaline stress effects. Gene sequencing revealed increased abundance of microbial genera, such as Tessaracoccus, Intrasporangium, Glutamicibacter, Agrobacterium, Saccharibacteria, Falsochrobactrum, Mesorhizobium, Bacillus, Ensifer, and Ornithinicoccus, under normal and saline–alkaline conditions, while functional changes in colony abundance were observed under different environments through PICRUST2 analysis. Bacillus subtilis demonstrated potential in enhancing rose growth and stress resistance under saline–alkaline conditions, affecting the regulation of the root microbial community. This study provides insights for improving soil conditions and enhancing plant adaptability in saline–alkaline regions.

An insight into variability and association among yield and its components under alkalinity and inland salinity stress in rice (Oryza sativa L.)

An experiment was conducted to study the variability, correlation, direct and indirect effects of yield and its attributing traits in 36 genotypes and to identify suitable parents for breeding of alkalinity and inland salinity stress tolerant lines at ARS, Kampasagar, Nalgonda during rabi, 2021-22. Analysis of variance revealed plenteous quantum of variation among the tested genotypes. Wide variation, high PCV, GCV, heritability and genetic advance were observed for seedling mortality, sterility percentage and number of filled grains per panicle. Number of productive tillers/hill and number of filled grains per panicle had a positive and significant correlation with yield whereas seedling mortality and sterility percentage had a negative and significant association with yield. Path analysis inferred that traits such as number of productive tillers/hill, number of grains per panicle and 1000 grain weight had a positive direct effect on yield. Hence,priority should be given to the above mentioned traits to develop alkalinity and inland saline tolerant rice cultivars. Lines CT118911, Sahel177, M202, and KPS10654 were found promising to be utilized as tolerant parents in the breeding programmes.

Hydrolysis of riboflavins in root exudates under iron deficiency and alkaline stress

Riboflavins are secreted under iron deficiency as a part of the iron acquisition Strategy I, mainly when the external pH is acidic. In plants growing under Fe-deficiency and alkaline conditions, riboflavins have been reported to accumulate inside the roots, with very low or negligible secretion. However, the fact that riboflavins may undergo hydrolysis under alkaline conditions has been so far disregarded. In this paper, we report the presence of riboflavin derivatives and products of their alkaline hydrolysis (lumichrome, lumiflavin and carboxymethylflavin) in nutrient solutions of Cucumis sativus plants grown under different iron regimes (soluble Fe-EDDHA in the nutrient solution, total absence of iron in the nutrient solution, or two different doses of FeSO4 supplied as a foliar spray), either cultivated in slightly acidic (pH 6) or alkaline (pH 8.8, 10 mM bicarbonate) nutrient solutions. The results show that root synthesis and exudation of riboflavins is controlled by shoot iron status, and that exuded riboflavins undergo hydrolysis, especially at alkaline pH, with lumichrome being the main product of hydrolysis.

Argon non-thermal plasma treatment promotes the development of rice (Oryza sativa L.) in saline alkali environments.

Soil salinization leads to a reduction in arable land area, which seriously endangers food security. Developing saline-alkali land has become a key measure to address the contradiction between population growth and limited arable land. Rice is the most important global food crop, feeding half of the world's population and making it a suitable choice for planting on saline-alkali lands. The traditional salt-alkali improvement method has several drawbacks. Currently, non-thermal plasma (NTP) technology is being increasingly applied in agriculture. However, there are few reports on the cultivation of salt/alkali-tolerant rice. Under alkaline stress, argon NTP treatment significantly increased the germination rate of Longdao 5 (LD5) rice seeds. In addition, at 15kV and 120s, NTP treatment significantly increased the activity of antioxidant enzymes such as catalase and SOD. NTP treatment induced changes in genes related to salt-alkali stress in rice seedlings, such as chitinase and xylanase inhibitor proteins, which increased the tolerance of the seeds to salt-alkali stress. This experiment has expanded the application scope of NTP in agriculture, providing a more cost-effective, less harmful, and faster method for developing salt-alkali-tolerant rice and laying a theoretical foundation for cultivating NTP-enhanced salt-alkali-tolerant rice.

Physiological and Biochemical Responses of Melilotus albus to Saline and Alkaline Stresses

Sweet clover (Melilotus albus) is a high-quality leguminous forage grass with salinity tolerance, drought tolerance, and cold hardiness. We selected four varieties of sweet clover with different sensitivities (061898, 061930, No. 55 white flower, and Ningxia white flower) and analyzed the effects of different concentrations of three sodium salts (Na2CO3, NaHCO3, and NaCl) on their physiology and biochemistry responses. Growth and development indexes (such as germination rate, root length, shoot length), chlorophyll content, osmotic regulators (proline, soluble sugar), malondialdehyde (MDA), superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were determined under saline–alkali stress. Seed germination and seedling growth of all four clover species were significantly inhibited under saline–alkali stress. During germination, seed germination rate, root length, and shoot length decreased with increasing saline and alkaline concentration. Under saline–alkali stress, chlorophyll content tended to increase and then decrease, cell damage and death increased, and malondialdehyde, soluble sugar, and proline content tended to increase and then decrease. Moreover, the activities of SOD, POD, and CAT all increased and then decreased. Under Na2CO3 stress, the decrease in chlorophyll content of the resistant variety 061898 was less than in the sensitive Ningxia white flower variety. As the concentrations of Na2CO3, NaHCO3, and NaCl increased, the maximum photochemical efficiency of PSII was significantly affected. The resistant 061898 is capable of maintaining higher photosynthetic efficiency. Furthermore, under treatments with the three kinds of saline–alkali solutions, cell damage and death for Ningxia white flower were greater than in 061898. For 061898, the increases in soluble sugar and proline content were greater and the increase in malondialdehyde content was less, while the antioxidant enzyme activities were higher than those in Ningxia white flower. All four sweet clover varieties had higher stress resistance with neutral than with alkaline salts. When stressed by medium to high saline–alkali concentrations, sweet clover seedlings had increased osmotic substance content, enhanced antioxidant enzyme activity, and regulated physiological metabolism. Additionally, sweet clovers regulated the expression of WRKY33, GH3, CYCD3, OXI1, MKK2, MYC2, JAZ, COI1, PYL, PP2C, TGA, and MPK3 to adapt to the saline–alkali environment and improve saline–alkali tolerance. Our analysis of the sweet clover salinity tolerance mechanism contributes to its further use and is of significant importance for addressing land salinization and promoting sustainable agricultural and pastoral practices in China.

Metabolomic characterization of alkali stress responses in rice

Soil alkalinity due to the accumulation of alkaline salts greatly impairs crop production. Despite advancements in crop stress response studies through metabolomic analyses, limited progress has been made in understanding cultivar differences in alkali tolerance due to the scarcity of suitable genetic material. This study aimed to characterize the metabolic responses to alkali stress in two rice cultivars, Kasalath (alkali-sensitive) and Gharib (alkali-tolerant), which were screened in an alkali soil field. A metabolomic analysis of the responses of hydroponically grown seedlings of both cultivars to alkaline and neutral salt stress was performed. Under alkali stress, the tolerant cultivar Gharib showed a significant accumulation of metabolites from the TCA organic acid and arginine synthesis pathways. This accumulation is consistent with observations in alkaliphilic wild grasses and may account for the superior alkali tolerance of Gharib. Although amino acids and nitrogen-containing metabolites, such as asparagine and allantoin, also accumulated under alkali stress, their accumulation was not specific to alkali stress, as previously reported. They accumulated similarly in both Kasalath and Gharib, suggesting that while these metabolites may alleviate alkali stress, they are unlikely to be responsible for cultivar differences in rice alkali tolerance.

CheV enhances the virulence of Salmonella Enteritidis, and the Chev-deleted Salmonella vaccine provides immunity in mice

BackgroundSalmonella enteritidis (SE) is a major zoonotic pathogen and causes infections in a variety of hosts. The development of novel vaccines for SE is necessary to eradicate this pathogen. Genetically engineered attenuated live vaccines are more immunogenic and safer. Thus, to develop a live attenuated Salmonella vaccine, we constructed a cheV gene deletion strain of SE (named ΔcheV) and investigated the role of cheV in the virulence of SE. First, the ability to resist environmental stress in vitro, biofilm formation capacity, drug resistance and motility of ΔcheV were analyzed. Secondly, the bacterial adhesion, invasion, intracellular survival assays were performed by cell model. Using a mouse infection model, an in vivo virulence assessment was conducted. To further evaluate the mechanisms implicated by the reduced virulence, qPCR analysis was utilized to examine the expression of the strain’s major virulence genes. Finally, the immune protection rate of ΔcheV was evaluated using a mouse model.ResultsCompared to C50336, the ΔcheV had significantly reduced survival ability under acidic, alkaline and thermal stress conditions, but there was no significant difference in survival under oxidative stress conditions. There was also no significant change in biofilm formation ability, drug resistance and motility. It was found that the adhesion ability of ΔcheV to Caco-2 cells remained unchanged, but the invasion ability and survival rate in RAW264.7 cells were significantly reduced. The challenge assay results showed that the LD50 values of C50336 and ΔcheV were 6.3 × 105 CFU and 1.25 × 107 CFU, respectively. After the deletion of the cheV gene, the expression levels of fimD, flgG, csgA, csgD, hflK, lrp, sipA, sipB, pipB, invH, mgtC, sodC, rfbH, xthA and mrr1 genes were significantly reduced. The live attenuated ΔcheV provided 100% protection in mice against SE infection.ConclusionAll the results confirmed that the deletion of the cheV gene reduces the virulence of SE and provides significant immune protection in mice, indicating that ΔcheV could be potential candidates to be explored as live-attenuated vaccines.

Acute stress response in gill of Pacific white shrimp Litopenaeus vannamei to high alkalinity

As a multifunctional organ, the gill plays a key role in the regulation of homeostasis. Excessive alkalinity in water can seriously threaten the survival of aquatic animals, however, the studies on the mechanism of alkalinity stress and adaption of crustacea are still limited. In this study, we aimed to investigate the mechanisms of acute alkalinity stress in the gill of Pacific white shrimp Litopenaeus vannamei by integrating physiological, histological, transcriptome, and metabolome analyses, which is an important aquaculture species of crustaceans. The physiological parameters related to homeostasis regulation, oxidative stress, material metabolism, and ammonia detoxification were significantly changed under acute alkalinity stress. There were a considerable number of genes and metabolites that were significantly changed in expression and level, which related to glycolysis/gluconeogenesis, fatty acid metabolism, amino acid metabolism, antioxidation, ammonia metabolism, and glycerophospholipid metabolism, In addition, the local structure of the gill was damaged and accompanied by cell apoptosis. The above results indicate that acute alkalinity stress can lead to damage to the gill structure, and affect its ion transport function, and the genes related to ion transport protein were upregulated. The antioxidant capacity in the gill was strengthened under acute alkalinity stress, at the same time, acute alkalinity stress would induce apoptosis in the gill. The glycogen catabolism in gill was significantly enhanced under acute alkalinity stress, while the lipid anabolism was inhibited, and the mechanism of the ornithine-urea cycle has been further mobilized to alleviate the inhibition of ammonia excretion and the accumulation of metabolic ammonia in the internal environment. In addition, the glycerophospholipid metabolism in the gill was disturbed under acute alkalinity stress. The current study will provide important clues for understanding the alkalinity stress mechanism of crustaceans, and provide an important theoretical basis for the optimization of shrimp aquaculture in saline-alkaline waters.

The Glycine soja cytochrome P450 gene GsCYP82C4 confers alkaline tolerance by promoting reactive oxygen species scavenging.

Recent studies have demonstrated the crucial role of Cytochrome P450 enzymes (CYPs) in the production of secondary metabolites, phytohormones and antioxidants in plants. However, their functional characterization specifically under alkaline stress remains elusive. CYP82C4 was the key gene screened from a family of wild soybean CYPs in our previous studies. The aim of this present study was to clone the Glycine soja GsCYP82C4 gene and characterize its functions in Arabidopsis and Glycine max. The results showed that the GsCYP82C4 gene displayed a high expression in different plant tissues at mature stages compared to young stages. Further, higher temporal expression of the GsCYP82C4 gene was noted at 6, 12 and 24 h time points after alkali treatment in leaves compared to roots. In addition, overexpression of GsCYP82C4 improved alkaline stress tolerance in Arabidopsis via increased root lengths and fresh biomass and strengthened the antioxidant defense system via a reduction in superoxide radicals in transgenic lines compared to wild type (WT) and atcyp82c4 mutants. Further, the expression levels of stress-related marker genes were up-regulated in GsCYP82C4 OX lines under alkali stress. The functional analysis of GsCYP82C4 overexpression in soybean displayed better hairy root growth, increased fresh weight, higher antioxidant enzyme activities and reduced lipid peroxidation rates in OX lines compared to the soybean WT (K599) line. In total, our study displayed positive roles of GsCYP82C4 overexpression in both Arabidopsis and Glycine max to alleviate alkaline stress via altering expression abundance of stress responsive genes, stronger roots, higher antioxidant enzyme activities as well as reduced rates of lipid peroxidation and superoxide radicals.

Ginger polysaccharide alleviates the effects of acute exposure to carbonate in crucian carp (Carassius auratus) by regulating immunity, intestinal microbiota, and intestinal metabolism

Alkaline stress poses a significant challenge to the healthy growth of fish. Ginger polysaccharide (GP) is one of the main active substances in ginger and has pharmacological effects, such as anti-oxidation and immune regulation. However, the physiological regulatory mechanism of GP addition to diet on alkalinity stress in crucian carp remains unclear. This study aimed to investigate the potential protective effects of dietary GP on antioxidant capacity, gene expression levels, intestinal microbiome, and metabolomics of crucian carp exposed to carbonate (NaHCO3). The CK group (no GP supplementation) and COG group (NaHCO3 stress and no GP supplementation) were set up. The GPCS group (NaHCO3 stress and 0.4% GP supplementation) was stressed for seven days. Based on these data, GP significantly increased the activities of total antioxidant capacity (T-AOC), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX), acid phosphatase (ACP), and alkaline phosphatase (AKP) in carp under alkalinity stress (p < 0.05) and decreased the activity of malon dialdehyde (MDA) (p < 0.05). GP restored the activity of GSH-PX, ACP, and AKP to CK levels. The expression levels of tumor necrosis factor β (TGF-β), tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), and interleukin 8 (IL-8) genes were decreased, and the expression levels of determination factor kappa-B (NF-κB) and interleukin 10 (IL-10) genes were increased (p < 0.05). Based on 16 S rRNA high-throughput sequencing, GP improved the changes in the intestinal microbial diversity and structural composition of crucian carp caused by NaHCO3 exposure. In particular, GP increased the relative abundance of Proteobacteria and Bacteroidetes and decreased the relative abundance of Actinobacteria. The metabolic response of GP to NaHCO3 exposed crucian carp guts was studied using LC/MS. Compared to the COG group, the GPCS group had 64 different metabolites and enriched 10 metabolic pathways, including lipid metabolism, nucleotide metabolism, and carbohydrate metabolism. The addition of GP to feed can promote galactose metabolism and provide an energy supply to crucian carp, thus alleviating the damage induced by alkalinity stress. In conclusion, GP can mitigate the effects of NaHCO3 alkalinity stress by regulating immune function, intestinal flora, and intestinal metabolism in crucian carp. These findings provide a novel idea for studying the mechanism of salt-alkali tolerance in crucian carp by adding GP to feed.

Comprehensive analysis of the aldehyde dehydrogenase gene family in Phaseolus vulgaris L. and their response to saline-alkali stress.

Aldehyde dehydrogenase (ALDH) scavenges toxic aldehyde molecules by catalyzing the oxidation of aldehydes to carboxylic acids. Although ALDH gene family members in various plants have been extensively studied and were found to regulate plant response to abiotic stress, reports on ALDH genes in the common bean (Phaseolus vulgaris L.) are limited. In this study, we aimed to investigate the effects of neutral (NS) and basic alkaline (AS) stresses on growth, physiological and biochemical indices, and ALDH activity, ALDH gene expression of common bean. In addition, We used bioinformatics techniques to analyze the physical and chemical properties, phylogenetic relationships, gene replication, collinearity, cis-acting elements, gene structure, motifs, and protein structural characteristics of PvALDH family members. We found that both NS and AS stresses weakened the photosynthetic performance of the leaves, induced oxidative stress, inhibited common bean growth, and enhanced the antioxidative system to scavenge reactive oxygen species. Furthermore, we our findings revealed that ALDH in the common bean actively responds to NS or AS stress by inducing the expression of PvALDH genes. In addition, using the established classification criteria and phylogenetic analysis, 27 PvALDHs were identified in the common bean genome, belonging to 10 ALDH families. The primary expansion mode of PvALDH genes was segmental duplication. Cis-acting elemental analysis showed that PvALDHs were associated with abiotic stress and phytohormonal responses. Gene expression analysis revealed that the PvALDH gene expression was tissue-specific. For instance, PvALDH3F1 and PvALDH3H1 were highly expressed in flower buds and flowers, respectively, whereas PvALDH3H2 and PvALDH2B4 were highly expressed in green mature pods and young pods, respectively. PvALDH22A1 and PvALDH11A2 were highly expressed in leaves and young trifoliates, respectively; PvALDH18B2 and PvALDH18B3 were highly expressed in stems and nodules, respectively; and PvALDH2C2 and PvALDH2C3 were highly expressed in the roots. PvALDHs expression in the roots responded positively to NS-AS stress, and PvALDH2C3, PvALDH5F1, and PvALDH10A1 were significantly (P < 0.05) upregulated in the roots. These results indicate that AS stress causes higher levels of oxidative damage than NS stress, resulting in weaker photosynthetic performance and more significant inhibition of common bean growth. The influence of PvALDHs potentially modulates abiotic stress response, particularly in the context of saline-alkali stress. These findings establish a basis for future research into the potential roles of ALDHs in the common bean.

Effects of drought-induced stress on nematode communities in aquatic and terrestrial habitats of the Nebraska Sandhills

IntroductionGlobal change events (e.g., worsening drought) are increasing environmental stress in a variety of terrestrial and aquatic habitats. The degree to which communities in soils and sediments are driven by temporal environmental changes across multiple habitat types from the same region is not clear.MethodsWe used nematodes, a common bioindicator of soil and sediment health, to determine how community diversity and composition are altered by rising alkalinity across lakes, shorelines, and prairies in the western Nebraska Sandhills. We sampled these three habitats from five lake basins spanning an alkalinity gradient (pH 7–11) across three years (2019, 2020, 2021). During our sampling, the Sandhills experienced a range of drought intensities, with 2019 being a wet year, followed by severe drought in 2020, and abnormally dry/moderate drought in 2021. To determine if diversity and composition of nematodes responded to increased alkalinity and drought-induced stress we used different modelling approaches, including Random Forest and pairwise comparisons.Results and discussionOverall, nematode diversity in lakes was most affected by increasing alkalinity over time, whereas in shorelines and prairies diversity was most reliant on bacterial diversity and potential nematode-nematode interactions. In comparison to shorelines and prairies, community composition in lakes was the least variable and consistently driven by pH and lake water levels. In contrast, compositions in the shorelines and prairies were more variable and explained at varying degrees by pH, year, lake basin, and climate-associated variables. In addition, relative abundance and compositional nature of select copious taxa were highly unpredictable, indicating potential instability in these habitats. Future research is necessary to address the ecologic stability of the Sandhills and determine where conservation efforts are most needed.

Structural Elucidation of Novel Degradation Impurities of Ibrutinib in Ibrutinib Tablets Using Preparative Chromatography, LCMS, HRMS and 2D NMR Techniques

Abstract Ibrutinib is an orally administered compound that functions as an irreversible covalent inhibitor of the Bruton tyrosine kinase, an essential element in multiple cellular processes including B-cell differentiation, proliferation, migration, survival and apoptosis. The compound has been found to demonstrate efficacy against a range of B-cell malignancies. The drug product is available in oral tablet and capsule formulations. The drug degradation profiles of tablets dosage form were assessed in accordance with regulatory guidelines. The results indicate that the drug substance is susceptible to alkaline and oxidative stress. The oxidation degradation led to the identification of three significant unknown degradation impurities. The three compounds were isolated through the application of preparative liquid chromatography, and their structures were determined using analytical techniques such as liquid chromatography–mass spectrometry, high-resolution mass spectrometry and nuclear magnetic resonance spectroscopy. Utilizing structural elucidation data, predictions were made regarding the composition of impurities, revealing them to be novel degradation impurities that bear structural resemblance to ibrutinib. Additionally, potential pathways for the formation of the impurities were proposed.

Integrated characterizations of intestinal bacteria and transcriptomics revealed the acute stress response to carbonate alkalinity in white shrimp Penaeusvannamei

The impact of carbonate alkalinity in saline-alkaline water on aquatic organisms, particularly Penaeus vannamei, a significant species in aquaculture, remains a critical area of study. To elucidate the acute response mechanisms of P. vannamei to elevated carbonate alkalinity environments, we utilized 16S rRNA gene and transcriptome sequencing technologies to analyze intestinal bacteria and gene expressions within various tissues. Our investigation revealed notable changes in specific intestinal bacterial OTUs, whose abundances varied preceding the overall bacterial community, indicating the sensitivity to carbonate alkalinity exposure. These shifts are accompanied by a simplification in bacterial networks and alterations in pathogenic OTUs, notably Aeromonas OTU. Concurrently, gene expression variations were observed across the hepatopancreas, gills, muscles, and intestines, with decreasing numbers of DEGs in the mentioned order. Annotation of these DEGs revealed enrichments in pathways related to transport, catabolism, immune responses, circulatory functions, and lipid metabolism. Notably, correlations between specific intestinal bacterial OTUs and gene expression shifts were identified across these tissues. Several OTUs, attributed to Rhizobiales, Saccharimonadales, Acidovora, and Aeromona, exhibited a correlation with DEGs in all four tissues, primarily associated with amino acid metabolism, signal transduction, and transport and catabolism pathways. Our study provides comprehensive insights into the dynamic responses of P. vannamei to elevated carbonate alkalinity stress. These findings contribute crucial knowledge for effective P. vannamei cultivation in saline-alkaline water, advancing our understanding in this field.