Enhancing Plant Stress Tolerance: The Role of LcWRKY40 Gene in Drought and Alkaline Salt Resistance in Tobacco and Yeast.

Leymus chinensis, a halophytic perennial grass belonging to the Poaceae family, thrives in saline-alkali grasslands and harbors a rich repository of resistance-related genetic resources. This study focused on deciphering the stress-responsive mechanisms of L. chinensis by conducting transcriptomic sequencing under NaHCO3 stress, which resulted in the annotation of a segment corresponding to the 51WRKY gene. The alkali-induced gene LcWRKY40 (QIG37591) was identified by phylogenetic analysis. Real-time quantitative PCR analysis was performed on L. chinensis plants subjected to PEG6000 and alkaline salt (NaHCO3) stress, and the results indicated that the LcWRKY40 gene was upregulated in both the leaves and roots. The localization of the LcWRKY40 protein was confirmed by the use of green fluorescent protein (GFP) fusion technology in transformed rice protoplast cells. The GAL4-driven transformation of the LcWRKY40 gene in INVScI yeast cells, which exhibited enhanced tolerance upon overexpression of the LcWRKY40 gene under mannitol and alkaline salt (NaHCO3) stress conditions. Under drought stress using mannitol, the fresh weight of Nicotiana tabacum overexpressing the LcWRKY40 gene was significantly higher than that of wild-type(WT) tobacco. Through drought and salt alkali stress, we found that overexpressed tobacco at different stages always outperformed the wild type in terms of fresh weight, SOD, MDA, and Fv/Fm. This study provides preliminary insights into the involvement of the LcWRKY40 gene in responding to drought and alkaline salt stresses, highlighting its role in enhancing plant resistance to drought and saline-alkaline conditions. These findings lay the foundation for future molecular breeding strategies aimed at improving grass resistance from different aspects.

Valpalf®: A New Nutraceutical Formulation Containing Bovine Lactoferrin That Exhibits Potentiated Biological Activity.

As a nutraceutical, bovine lactoferrin (bLf), an iron-binding glycoprotein involved in innate immunity, is gaining elevated attention for its ability to exert pleiotropic functions and to be exceptionally tolerated even at high dosages. Some of bLf's activities, including its anti-inflammatory and antioxidant, are tightly linked to its ability to both chelate iron and enter inside the cell nucleus. Here, we present data about Valpalf®, a new formulation containing bLf, sodium citrate, and sodium bicarbonate at a molar ratio of 10-3. In the present study, Valpalf® exhibits superior iron-binding capacity, resistance to tryptic digestion, and a greater capacity to accumulate into the nucleus over time when compared to the native bLf alone. In agreement, Valpalf® effectively reduces interleukin(IL)-6 levels in lipopolysaccharide-stimulated macrophages and modulates the expression of antioxidant enzymes, such as superoxide dismutase 1 and 2, in phorbol-12-myristate-13-acetate-stimulated monocytes. Of note, this potentiated bioactivity was corroborated in a retrospective study on the treatment of anemia of inflammation in hereditary thrombophilic pregnant and non-pregnant women, demonstrating that Valpalf® improves hematological parameters and reduces serum IL-6 levels to a higher extent than bLf alone.

Effects of sodium bicarbonate solution on hypergravity-induced Fos expression in neurons of the amygdala in rats: Implication of sodium bicarbonate therapy for vertigo.

OBJECTIVE: In Japan, intravenous injection of a 7 % solution of sodium bicarbonate (NaHCO3) had been originally developed to inhibit motion sickness and then have long been used to treat vertigo. Previously, we reported that Fos-positive neurons appear in the amygdala after hypergravity stimulation in rats. In the present study, we examined whether injection of 7 % NaHCO3 inhibits hypergravity-induced Fos expression in the neurons in the central nucleus of the amygdala in rats. METHODS: Rats were exposed to 2 G hypergravity in an animal centrifuge device for 3 h. A solution of 7 % NaHCO3 at a dose of 4 mM/kg was injected intraperitoneally before 2 G hypergraviy. Fos-positive neurons in the amygdala were stained immunohistochemically. RESULTS: The number of Fos-positive neurons in the central nucleus of the amygdala was significantly increased after 2 G hypergravity in rats that received no drugs or saline, compared to that in rats exposed only to the noise of the centrifuge and received 7 % NaHCO3 solution. The number of Fos-positive neurons in the central nucleus of the amygdala after 2 G hypergravity was significantly decreased in rats that received 7 % NaHCO3 solution, compared to that in rats that received no drugs or saline. CONCLUSION: Since Fos expression is a marker of activated neurons, the present findings suggest that hypergravity activates the amygdala and that administration of 7 % NaHCO3 suppresses hypergravity-induced activation of the amygdala. Hypergravity disturbs spatial orientation to produce motion sickness and the amygdala is involved in fear response. Recently, Ziemann et al. suggested that fear-evoking stimuli reduce the pH in the amygdala to activate it, leading to induction of fear behavior and that administering HCO3- attenuates fear behavior [Cell 2009; 139: 1012-1021]. Therefore, it is possible that hypergravity reduces the pH in the amygdala to activate it, thereby inducing the fear associated with motion sickness and that administration of 7 % NaHCO3 increases the brain pH thereby suppressing hypergravity-induced activation of the amygdala and inhibiting the fear associated with motion sickness. In patients with vertigo, 7 % NaHCO3 therapy may increase the brain pH thereby suppressing the activation of the amygdala and inhibiting the fear associated with vertigo to elicit a beneficial clinical effect.

Cationic liposomes as a drug-free system for efficient anticancer therapy by intracytoplasmic delivery of sodium bicarbonate.

Developing the delivery systems with high therapeutic efficacy and low side effects is of great interest and significance for anticancer therapy. Compared to the high cost in synthesizing new chemotherapeutic drugs, exploring the anticancer potentials of existing chemicals is more convenient and efficient. Sodium bicarbonate (BC), a simple inorganic salt, has shown its tumor inhibition capacity via regulating the acidity of tumor microenvironment. However, the effects of intracytoplasmic BC on tumor growth and the potentials of BC to serve as an anticancer agent are still unknown. Herein, we developed a BC-loaded cationic liposome system (BC-CLP) to deliver BC into the cytosol of cancer cells. The in vitro studies showed that the BC-CLP containing 1% BC (w/v) had a size of 112.9 nm and a zeta potential of 19.1 mV, which reduced the viability of the model cancer cells (human oral squamous cell carcinoma HSC-3 cells) to 13.7%. In contrast, the neutral BC-LP caused less than 50% viability reduction. We further found that BC-CLP released BC directly into cytoplasm via membrane fusion pathway rather than endocytosis, leading to the remarkable increase of cytosolic pH, which may contribute to the anticancer effect of BC-CLP. Our findings indicate that BC-CLP is a potential system for high-efficiency cancer therapy without causing drug-related side effects or resistance.

EFFECT OF MIR-21-3P ON INTESTINAL INJURY IN RATS WITH TRAUMATIC HEMORRHAGIC SHOCK RESUSCITATED WITH THE SODIUM BICARBONATE RINGER'S SOLUTION.

ABSTRACT: Background : This study aims to determine the impact and mechanism of miR-21-3p on intestinal injury and intestinal glycocalyx during fluid resuscitation in traumatic hemorrhagic shock (THS), and the different impacts of sodium lactate Ringer's solution (LRS) and sodium bicarbonate Ringer's solution (BRS) for resuscitation on intestinal damage. Methods : A rat model of THS was induced by hemorrhage from the left femur fracture. The pathological changes of intestinal tissues and glycocalyx structure were observed by hematoxylin-eosin staining and transmission electron microscope. MiR-21-3p expression in intestinal tissues was detected by real-time quantitative polymerase chain reaction. The expression of glycocalyx-, cell junction-, and PI3K/Akt/NF-κB signaling pathway-related proteins was analyzed by western blot. Results : MiR-21-3p expression was increased in THS rats, which was suppressed by resuscitation with BRS. BRS or LRS aggravated the intestinal injury and damaged intestinal glycocalyx in THS rats. The expression of SDC-1, HPA, ß-catenin, MMP2, and MMP9 was upregulated, the expression of E-cad was downregulated, and the PI3K/Akt/NF-κB signaling pathway was activated in THS rats, which were further aggravated by BRS or LRS. The adverse effect of LRS was more serious than BRS. MiR-21-3p overexpression deteriorated the injury of intestinal tissues and intestinal glycocalyx; increased the expression of SDC-1, HPA, ß-catenin, MMP2, and MMP9 while decreasing E-cad expression; and activated the PI3K/Akt/NF-κB signaling pathway in BRS-resuscitated THS rats. Conclusion : MiR-21-3p aggravated intestinal tissue injury and intestinal glycocalyx damage through activating PI3K/Akt/NF-κB signaling pathway in rats with THS resuscitated with BRS.

Effects of acute and multi-day low-dose sodium bicarbonate intake on high-intensity endurance exercise performance in male recreational cyclists.

PURPOSE: This study aimed to compare the effects of acute and multi-day low-dose sodium bicarbonate (SB) intake on high-intensity endurance exercise performance. METHODS: In a randomized, double-blind, cross-over design, twelve recreational male cyclists (age: 31.17 ± 4.91 years; V ˙ O2peak: 47.98 ± 7.68 ml·kg-1·min-1) completed three endurance performance tests following acute SB (ASB, 0.2 g·kg-1 SB), multi-day SB (MSB, 0.2 g·kg-1·day-1 SB for four days), and placebo (PLA) intake. The high-intensity endurance performance was assessed with a cycling exercise test, wherein participants cycled on a bicycle ergometer at 95% of the predetermined anaerobic threshold for 30 min, followed by a time-to-exhaustion test at 110% of the anaerobic threshold. Data were analyzed using one-way and two-way repeated-measures ANOVA. RESULTS: Significant main effects of supplementation protocol were evident in pre-exercise bicarbonate concentrations (F = 27.93; p < 0.01; partial eta squared (η2) = 0.72; false discovery rate (FDR)-adjusted p value = 0.001). Prior to performance test, blood bicarbonate concentrations were significantly higher in MSB (25.78 ± 1.63 mmol·L-1 [95% CI 26.55-28.44] (p < 0.001; FDR-adjusted p value = 0.001)) and ASB (27.49 ± 1.49 mmol·L-1 [95% CI 24.75-26.81] (p < 0.001; FDR-adjusted p value = 0.007)) compared to PLA (23.75 ± 1.40 mmol·L-1 [95% CI 22.86 to 24.64]). Time-to-exhaustion increased in MSB (54.27 ± 9.20 min [95% CI 48.43-60.12]) compared to PLA (49.75 ± 10.80 min [95% CI 42.89-56.62]) (p = 0.048); however, this increase in MSB did not reach the significance threshold of 1% FDR (FDR-adjusted p value = 0.040). No significant difference was noted in exhaustion times between ASB (51.15 ± 8.39 min [95% CI 45.82-56.48]) and PLA (p > 0.05). CONCLUSION: Both acute and multi-day administration of low-dose SB improves buffering system in cyclists; nevertheless, neither intervention demonstrates sufficient efficacy in enhancing high-intensity endurance performance.

The conserved evolution of plant H+-ATPase family and the involvement of soybean H+-ATPases in sodium bicarbonate stress responses.

Plant plasma membrane (PM) H+-ATPases are essential pumps involved in multiple physiological processes. They play a significant role in regulating pH homeostasis and membrane potential by generating the electrochemical gradient of the proton across the plasma membrane. However, information on soybean PM H+-ATPase is still limited. In this study, we conducted the evolutionary analysis of PM H+-ATPases in land plants and investigated the subfamily classification and whole genome duplication of PM H+-ATPases in angiosperms. We further characterized the extremely high conservation of the soybean PM H+-ATPase family in terms of gene structure, domain architecture, and protein sequence identity. Using the yeast system, we confirmed the highly conserved biochemical characteristics (14-3-3 binding affinity and pump activity) of soybean PM H+-ATPases and their conserved function in enhancing tolerance to high pH and NaHCO3 stresses. Meanwhile, our results also revealed their divergence in the transcriptional expression in different tissues and under sodium bicarbonate stress. Finally, the function of soybean PM H+-ATPases in conferring sodium bicarbonate tolerance was validated using transgenic Arabidopsis. Together, these results conclude that the soybean PM H+-ATPase is evolutionarily conserved and positively regulates the response to sodium bicarbonate stress.

Sodium bicarbonate treatment for QRS widening in bupropion overdoses.

INTRODUCTION: Bupropion cardiotoxicity widens QRS complexes by inhibiting cardiac gap junctions. Sodium bicarbonate is the standard treatment for QRS widening from sodium channel blockade, but its effect on QRS widening in bupropion cardiotoxicity is not well-studied. METHODS: This is a retrospective cohort study of bupropion overdoses from 10 hospitals between January 2010 and June 2022. Patients with documented administration of sodium bicarbonate and QRS duration > 100 milliseconds on pre-bicarbonate electrocardiogram were included. Patients with no electrocardiogram within four hours of treatment or with baseline pre-overdose wide QRS and < 10 milliseconds widening from baseline were excluded. The primary outcome was a change in QRS duration between the pre-bicarbonate electrocardiogram and the first electrocardiogram after initial bicarbonate administration. Secondary outcomes included prevalence of post-bicarbonate QRS < 100 milliseconds, change in electrocardiogram intervals after total bicarbonate administration, and change in metabolic parameters and hemodynamics. Wilcoxon signed-rank testing was performed on the primary outcome. Linear regression modeling was performed to test for an association between change in QRS and bicarbonate dosing. RESULTS: Thirteen patients were included for final analysis. The median age was 32 years, and 54% were male. Six patients developed seizures; one developed ventricular tachycardia, and four received vasopressors. The median QRS and QTc pre-bicarbonate were 116 and 495 milliseconds, respectively. The median change in QRS duration was -2.0 milliseconds, which was not statistically significant (P = 0.42). The median bicarbonate dose administered before the first post-bicarbonate electrocardiogram was 100 milliequivalents. We did not identify an association between QRS change and bicarbonate dosing (P = 0.9, R-squared = 0.001). No patient had a QRS duration < 100 milliseconds after the initial bicarbonate dose. There was minimal change in QTc, electrolytes, heart rate, or blood pressure; alkalemia post-bicarbonate was achieved in eight patients. CONCLUSION: Sodium bicarbonate did not significantly decrease QRS duration in this small retrospective cohort of bupropion overdoses.

Sodium Bicarbonate (NaHCO3) Increases Growth, Protein and Photosynthetic Pigments Production and Alters Carbohydrate Production of Spirulina platensis.

Improving the biochemical status of Spirulina platensis will enhance the functional properties of this microalgae. The present study investigated the effects of adding NaHCO3 to the culture medium on the growth rate and biochemical composition, particularly the coproduction of proteins, carbohydrates, and photosynthetic pigments of S. platensis. Spirulina platensis was grown in different NaHCO3 concentrations (0-16 g L-1). NaHCO3 positively affected the biomass production. The growth of S. platensis and biochemical compound content increased with an increase in the NaHCO3 concentration. The microalgae biomass grown on NaHCO3 also contained higher amounts of protein (64.20 ± 4.18% w w-1) and photosynthetic pigments (phycocyanin and chlorophyll a, b, and total). Protein productivity was especially enhanced by approximately 6-25% (from 0.006 ± 0.0030 to 0.025 ± 0.0031 mg L-1 day-1) with the addition of NaHCO3 compared to the control. In contrast, the content of carbohydrates and antioxidant compounds (phenolic, polyphenol oxidase, and peroxidase activities) decreased with culture age and an increase in the NaHCO3 concentration. These results suggest that S. platensis uses NaHCO3 as a carbon source for photosynthesis, biomass production, and acts as a metabolic energy carrier toward the synthesis of proteins and photosynthetic pigments, which are more energy-consuming metabolites than carbohydrates. The addition of NaHCO3 to the culture media is a potentially useful strategy toward improving the protein and photosynthetic pigment productivity of S. platensis.

The effects of enteric-coated sodium bicarbonate supplementation on 2 km rowing performance in female CrossFit® athletes.

PURPOSE: Sodium bicarbonate (SB) supplementation can improve exercise performance, but few studies consider how effective it is in female athletes. The aim of the study was to establish the effect of individually timed pre-exercise SB ingestion on 2 km rowing time trial (TT) performance in female athletes. METHODS: Eleven female CrossFit® athletes (mean ± SD age, 29 y ± 4 y, body mass, 64.5 kg ± 7.1 kg, height, 1.7 m ± 0.09 m, peak oxygen uptake [VO2peak], 53.8 ± 5.7 mL·kg-1∙min-1). An initial trial identified individual time-to-peak [HCO3-] following enteric-coated 0.3 g·kg-1 BM SB ingestion. Participants then completed a 2 km TT familiarisation followed by a placebo (PLA) or SB trial, using a randomised cross-over design. RESULTS: The ingestion of SB improved rowing performance (514.3 ± 44.6 s) compared to the PLA (529.9 ± 45.4 s) and FAM trials (522.2 ± 43.1 s) (p = 0.001, pη2 = 0.53) which represents a 2.24% improvement compared to the PLA. Individual time-to-peak alkalosis occurred 102.3 ± 22.1 min after ingestion (range 75-150 min) and resulted in increased blood [HCO3-] of 5.5 ± 1.5 mmol⋅L-1 (range = 3.8-7.9 mmol⋅L-1). The change in blood [HCO3-] was significantly correlated with the performance improvement between PLA and SB trials (r = 0.68, p = 0.020). CONCLUSIONS: Ingesting a 0.3 g·kg-1 BM dose of enteric-coated SB improves 2 km rowing performance in female athletes. The improvement is directly related to the extracellular buffering capacity even when blood [HCO3-] does not change ≥ 5.0 mmol⋅L-1.