Saline Treatment Research Articles

Splenic gene expression of cytokines at multiple time points following lipopolysaccharide challenge in layers.

To investigate inflammatory responses to lipopolysaccharide (LPS) injection in layers. 33 40-week-old laying hens were used. 30 laying hens were divided into 2 groups: the first group was injected with 8 mg/kg LPS, while the second group was injected with sterile saline. At the start of the study, 3 birds served as a baseline and were used as the time 0 controls for both the saline and LPS-treated groups. Blood and spleen tissues were collected at 0 (before) and 1, 2, 3, 4, and 6 hours after injection. LPS administration increased splenic mRNA levels of IL-1β, IL-2, IL-6, IL-8, IL-10, interferon-γ, and tumor necrosis factor-α (P < .001) and serum IL-6 levels (P < .01) compared to saline injection. The mRNA expression of most cytokine genes increased rapidly toward peak values within 2 hours after the LPS injection, and then the difference between the saline and LPS treatments got smaller as time went on; serum IL-6 reached its highest concentration 2 hours after LPS administration. The magnitude of LPS-induced upregulation of gene expression was the highest for IL-6, followed by IL-1β and IL-8, and tumor necrosis factor-α was the least affected. The temporal and quantitative profile of these inflammatory mediators generated from this study provides valuable information in identifying the optimal time window and appropriate biomarkers for LPS-induced inflammation, which has significant implications in evaluating the effects of interventions on the immune system of chickens.

Induction of TNFα by flavin adenine dinucleotide (FADH) protects neonatal C57Bl6 lungs from high oxygen induced lung injury

Background: Bronchopulmonary dysplasia (BPD) is a serious lung disease caused by oxidative stress, predominately affecting premature infants that require prolonged oxygen support. The co-factor flavin adenine dinucleotide (FADH) facilitates glutathione reductase (GR) enzymatic activity and increases the bioavailability of the antioxidant glutathione (GSH). As such, we hypothesized that intranasal delivery of FADH can improve redox homeostasis and attenuate lung injury following chronic high oxygen supplementation (0.85% FiO2) by altering pro-inflammatory signal transduction pathways in a C57Bl6 newborn mouse model for BPD. Methods: We used a prolonged hyperoxia-induced lung injury rodent model to recapitulate the clinical and histological changes present in BPD. Newborn mice were housed in FiO2 85% (hyperoxic) or 21% O2 (normoxic) from day of birth until post-natal day (PN) 14. FADH (7 μM) or saline (control) treatments were administered daily via intranasal insufflation on PN 14-21 in a final volume of 1.5μL/g body weight. BALF and lung tissue samples were collected and evaluated for redox stress, lung injury/development, and inflammation using standard immunohistological techniques, morphometric analysis, and standard molecular assays. BALF cytokines were measured using a flow cytometric approach (BioLegend). Two-dimensional cell migration assays were tracked via microscopy daily until 100% confluence reached; the ½ maximal peak values were evaluated to compare rate of wound closure following very low (2.5 ng/mL) TNFα or IL12 treatment. Transepithelial resistance and potential differences were measured using an EVOM device (World Precision Instrument) across human small airway epithelial cells, and equivalent short circuit currents (Isc) calculated in accordance with Ohm’s Law, where V=IR. Single data comparisons were performed using paired Student’s t-tests. Multiple comparisons were performed using ANOVA followed by post-hoc testing in Sigma Plot. Results: FADH protected neonatal lungs from high oxygen induced oxidative stress: GSH/GSSG Eh (a measure of oxidative stress) improved from -168.77 mV ± 3.64 mV to -179.10 mV ± 1.85 mV (n=5 BALF measurements). FADH also improved lung injury scores from 0.187±0.038 to 0.03±0.014 (p=0.005), decreased neutrophil migration (p&lt;0.001), and increased macrophages (p&lt;0.001) when compared to age-matched untreated pups. FADH-mediated increase in TNFα (and not IL12) significantly increased the rate of lung epithelial cell wound closure from [2.27±0.07 to 1.97±0.1 average days for ½ complete wound closure, n=6, p=0.01] and increased Isc from 6.5±.21 to 9.2±1.0 μA/cm2, n=3, p=0.03. Conclusions: Our findings indicate that FADH protects the preterm lung from hyperoxic injury by 1) increasing the bioavailability of GSH and 2) via TNFα signaling to increase the rate of wound closure and repair of epithelial cell function. Significances: The mechanisms identified through this study could be targeted as a potential adjunct therapy for BPD patients to attenuate high oxygen induced lung injury. Funding: Supported by R01HL137033 (MH). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Genome-wide identification, characterization and functional prediction of the SRS gene family in sesame (Sesamum indicum L.)

Sesame (Sesamum indicum L.) is an ancient oilseed crop of the Pedaliaceae family with high oil content and potential health benefits. SHI RELATED SEQUENCE (SRS) proteins are the transcription factors (TFs) specific to plants that contain RING-like zinc finger domain and are associated with the regulation of several physiological and biochemical processes. They also play vital roles in plant growth and development such as root formation, leaf development, floral development, hormone biosynthesis, signal transduction, and biotic and abiotic stress responses. Nevertheless, the SRS gene family was not reported in sesame yet. In this study, identification, molecular characterization, phylogenetic relationship, cis-acting regulatory elements, protein-protein interaction, syntenic relationship, duplication events and expression pattern of SRS genes were analyzed in S. indicum. We identified total six SiSRS genes on seven different linkage groups in the S. indicum genome by comparing with the other species, including the model plant Arabidopsis thaliana. The SiSRS genes showed variation in their structure like 2–5 exons and 1–4 introns. Like other species, SiSRS proteins also contained ‘RING-like zinc finger’ and ‘LRP1’ domains. Then, the SiSRS genes were clustered into subclasses via phylogenetic analysis with proteins of S. indicum, A. thaliana, and some other plant species. The cis-acting regulatory elements analysis revealed that the promoter region of SiSRS4 (SIN_1011561) showed the highest 13 and 16 elements for light- and phytohormone-responses whereas, SiSRS1 (SIN_1015187) showed the highest 15 elements for stress-response. The ABREs, or ABA-responsive elements, were found in a maximum of 8 copies in the SiSRS3 (SIN 1009100). Moreover, the available RNA-seq based expression of SiSRS genes revealed variation in expression patterns between stress-treated and non-treated samples, especially in drought and salinity conditions in. S. indicum. Two SiSRS genes like SiSRS1 (SIN_1015187) and SiSRS5 (SIN_1021065), also exhibited variable expression patterns between control vs PEG-treated sesame root samples and three SiSRS genes, including SiSRS1 (SIN_1015187), SiSRS2 (SIN_1003328) and SiSRS5 (SIN_1021065) were responsive to salinity treatments. The present outcomes will encourage more research into the gene expression and functionality analysis of SiSRS genes in S. indicum and other related species.

The Impacts of Osmotic Stress During the Critical Window of Development on Protein and Histone PTM Alterations in California grunion

The California Grunion, Leuresthes tenuis, can experience a broad range of salinities during their early life stages, with their nursery grounds measuring salinities of 21 to 42ppt under normal conditions. Due to the unique subterrestrial incubation of their eggs within the sand of California beaches, it is not unlikely for this marine fish to hatch in non-seawater conditions, whereafter the larvae may transition to relatively diluted estuaries or concentrated harbors to develop as juveniles for several months. Consequently, they are an ideal system to study the impacts of ecologically relevant salinity stress on the alteration of proteins and histone post-translational modifications (PTMs) during early development. The study of the relationship between the critical window of development (CWD), in which sexual bipotential is lost, and the longevity of protein landscape and histone PTM changes has not been widely explored. The CWD is of interest as this is when the gametes of this species become set. Therefore, we hypothesize that histone PTM changes that occur during this time may thus be a form of heritable epigenetics. Thus, hypo-and-hyperosmotic exposures of L. tenuis hatchlings during this window, and past it, attempt to identify if the CWD is relevant to the presence and persistence of protein and histone PTM changes under salinity stress. We hypothesize that exposure to osmotic stress during the CWD will result in induced compensatory mechanisms in the protein landscape of the California grunion larvae, as well as persistent alterations in the histone PTMs in the larvae. To elucidate the relationship between these factors, California grunion larvae were exposed in replicate to one of three chronic salinity treatments (16ppt, 32ppt, or 40ppt) for the duration of their CWD (68dph), with 16ppt and 40ppt inducing hypoosmotic and hyperosmotic stress respectively. Chronic post-CWD exposures and recovery times until 98dph were used to determine whether the CWD or prolonged salinity exposures were key to alterations in the histone PTM and protein landscape of the larvae. After each timepoint, L. tenuis were culled and processed for histone PTM enrichment and proteomic analysis. At the time of culling, there were no significant differences in survivorship between replicates and experimental groups, with statistically significant different deaths between groups occurring only shortly after hatching. Skyline and MSstats were used in the analysis of the statistical significance of differential regulation of proteins and histone PTMs under stress conditions versus the controls. Of special interest is the osmoregulatory mechanisms involved in the myo-inositol biosynthesis pathway, with analysis focusing on the abundance of histone acetylation, amidation, ubiquitylation, and 4-hydroxynonelation in histone PTMs, as the accumulations of these have been found to be correlated with osmotic stress in some fish tissues. Funding provided by NSF IOS-2209383 and NSF GRFP. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Recurrent Ethanol Treatment does not Produce a Cumulative Effect on Plasma Leptin Levels in Rats

Background: Hypoleptinemia is a potent orexigenic signal. Chronic ethanol (EtOH) consumption is associated with weight gain in humans, and acute ethanol treatment leads to reductions in leptin concentrations in both fasted humans and rats. The objective of this study was to determine whether short-term recurrent ethanol treatment has a cumulative effect on plasma leptin levels in rats, and whether treatment in the fed or fasted state influences the leptin lowering effect of EtOH. We hypothesized that recurrent EtOH treatment would have a cumulative effect on leptin levels, and that these effects would be more pronounced in the fed state, when leptin levels are highest. Methods: Two separate groups of male Sprague Dawley rats were exposed to either oral or intravenous (IV) treatment with EtOH (2g/kg via oral gavage or 0.067 g/kg/min IV for 30 min; n=7/group) on three consecutive days. Each rat served as its own control and received saline treatment either 7 d prior to or 7 d following exposure to EtOH. The oral group was fasted 12-14 h (overnight) prior to treatment, while the IV group was not fasted. Plasma leptin and EtOH levels were measured each day at baseline and 3 h following EtOH treatment. Data are expressed as mean ± SEM. A two-way repeated measures ANOVA was used to test for differences in plasma leptin and EtOH levels due to treatment (saline vs. EtOH) or day (cumulative effect of treatment). Results: Post-treatment ethanol levels in the oral and IV groups (55 ± 6.9 and 45 ± 6.3 mM, respectively) were higher than saline-treated rats (p&gt;0.001). Baseline leptin levels in the oral treatment (fasted) group were 3.4 ± 0.3 and 3.9 ± 0.2 ng/mL under saline and leptin treated conditions, respectively. There was a significant main effect of treatment but not of day in the oral group (p=0.04 and p=0.38, respectively). In the oral group, average daily change in leptin levels 3 h post treatment in saline and leptin treated rats were -0.18 ± 0.05 and -0.72 ± 0.1 ng/mL, respectively. Baseline leptin levels in the IV group (fed) were 7.8 ± 0.3 and 6.7 ± 0.1 ng/mL, in saline and leptin treated rats respectively. There was no significant main effect of treatment or day in the IV group (p=0.17 and p=0.41, respectively). In the IV group, average daily changes in leptin levels 3 h following treatment in saline and leptin treated rats were -1.7 ± 0.2 and -1.3 ± 0.3 ng/mL, respectively. Conclusion: Although we were able to replicate previous work indicating reduced leptin levels following acute EtOH treatment in fasted rats, our results do not support a cumulative leptin-lowering effect of recurrent EtOH treatment. Furthermore, IV EtOH treatment in the fed state did not have any impact on leptin levels. This suggests that the leptin-lowering effects of EtOH may either require the fasted state or may require ingestion and exposure of the GI system to EtOH. NIH R01 DK131165-01A1. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Enhanced Sweat Biosensing with Thread-Embedded Microfluidic Devices.

BACKGROUND This study explored the integration of conductive threads into a microfluidic compact disc (CD), developed using the xurographic method, for a potential sweat biosensing platform. MATERIAL AND METHODS The microfluidic CD platform, fabricated using the xurographic method with PVC films, included venting channels and conductive threads linked to copper electrodes. With distinct microfluidic sets for load and metering, flow control, and measurement, the CD's operation involved spinning for sequential liquid movement. Impedance analysis using HIOKI IM3590 was conducted for saline and artificial sweat solutions on 4 identical CDs, ensuring reliable conductivity and measurements over a 1 kHz to 200 kHz frequency range. RESULTS Significant differences in |Z| values were observed between saline and artificial sweat treatments. 27.5 μL of saline differed significantly from 27.5 μL of artificial sweat, 72.5 μL of saline from 72.5 μL of artificial sweat, and 192.5 μL of saline from 192.5 μL of sweat. Significant disparities in |Z| values were observed between dry fibers and Groups 2, 3, and 4 (varying saline amounts). No significant differences emerged between dry fibers and Groups 6, 7, and 8 (distinct artificial sweat amounts). These findings underscore variations in fiber characteristics between equivalent exposures, emphasizing the nuanced response of the microfluidic CD platform to different liquid compositions. CONCLUSIONS This study shows the potential of integrating conductive threads in a microfluidic CD platform for sweat sensing. Challenges in volume control and thread coating degradation must be addressed for transformative biosensing devices in personalized healthcare.

In situ pulmonary mucus hydration assay using rotational and translational diffusion of gold nanorods with polarization-sensitive optical coherence tomography.

Assessing the nanostructure of polymer solutions and biofluids is broadly useful for understanding drug delivery and disease progression and for monitoring therapy. Our objective is to quantify bronchial mucus solids concentration (wt. %) during hypertonic saline (HTS) treatment in vitro via nanostructurally constrained diffusion of gold nanorods (GNRs) monitored by polarization-sensitive optical coherence tomography (PS-OCT). Using PS-OCT, we quantified GNR translational () and rotational () diffusion coefficients within polyethylene oxide solutions (0 to 3wt. %) and human bronchial epithelial cell (hBEC) mucus (0 to 6.4wt. %). Interpolation of and data is used to develop an assay to quantify mucus concentration. The assay is demonstrated on the mucus layer of an air-liquid interface hBEC culture during HTS treatment. In polymer solutions and mucus, and monotonically decrease with increasing concentration. is more sensitive than to changes above 1.5wt. % of mucus and exhibits less intrasample variability. Mucus on HTS-treated hBEC cultures exhibits dynamic mixing from cilia. A region of hard-packed mucus is revealed by measurements. The extended dynamic range afforded by simultaneous measurement of and of GNRs using PS-OCT enables resolving concentration of the bronchial mucus layer over a range from healthy to disease in depth and time during HTS treatment in vitro.

Morphological and Physiological Responses of Different Cotton Genotypes Primed with Salicylic Acid Under Salinity Conditions

This study was conducted as both petri dishes and pot experiments on four different salt-sensitive cotton genotypes (Laser, May 505, May 455 and Selin) in order to investigate the role of exogenous salicylic acid applications in reducing the effects of salt stress. Six saline treatments; 0, 30, 60, 90, 120 mM NaCl were used. Each group divided into three sub-groups (hydo-primed control, 0.5 mM and 1.0 mM SA) on the basis of seed priming treatments. They were applied in three replications according to the randomized block design. In all genotypes, 90 mM and 120 mM salt stress negatively affected germination and seedling development. In salt stress up to 60 mM, it was recommended to May 505 and Selin genotypes with 0.5 mM salicylic acid pre-application to the seeds.

Saline, chlorhexidine, and povidone-iodine alone or in combination with iodine povacrylex are effective antiseptics in chickens (Gallus gallus domesticus).

To evaluate the topical antiseptic activity of saline, chlorhexidine (CHX), and povidone-iodine (PI) scrubs on the skin of chickens with or without the addition of DuraPrep (DP). 7 healthy adult Orpington hens (Gallus gallus domesticus). The right apterium corporale laterale was swabbed for standard aerobic bacterial culture and colony-forming unit (CFU) determination. The apterium was divided into 3 areas and treated with sterile saline, CHX, or PI. Samples were collected by swabbing each area before and after additional treatment with DP. CFU's were counted after 48 hours of incubation. Statistical analysis was performed using a linear mixed model with a continuous outcome. Compared to saline, CHX and PI treatment without DP decreased CFU count by 119 (95% CI, 55 to 183; P < .001) and 123 (95% CI, 58 to 187; P < .001), respectively. The application of DP after CHX and PI further decreased CFU counts by 6 (P = .01) and 9 (P = .01), respectively. DP after saline treatment decreased counts by 128 CFU (95% CI, 63 to 192; P < .001). No significant difference was detected between saline, PI, or CHX after DP application (-1.0 CFU; 95% CI, 63.4 to -65.4; P = .98 for both PI and CHX). CHX or PI provided greater reductions in bacterial CFU than saline, and all combinations with DP provided similar results. No notable cutaneous reactions were detected at any point. This data suggests that a scrub protocol including CHX or PI with DP is acceptable in surgical site preparation of chickens.

Exploring Iodide and Hydrogen Sulfide as ROS Scavengers to Delay Acute Rejection in MHC-Defined Vascularized Composite Allografts.

Vascularized composite allografts (VCA) face ischemic challenges due to their limited availability. Reperfusion following ischemia triggers oxidative stress and immune reactions, and scavenger molecules could mitigate ischemia-reperfusion injuries and, therefore, immune rejection. We compared two scavengers in a myocutaneous flap VCA model. In total, 18 myocutaneous flap transplants were performed in Major histocompatibility complex (MHC)-defined miniature swine. In the MATCH group (n = 9), donors and recipients had minor antigen mismatch, while the animals were fully mismatched in the MISMATCH group (n = 9). Grafts were pretreated with saline, sodium iodide (NaI), or hydrogen sulfide (H2S), stored at 4 °C for 3 h, and then transplanted. Flaps were monitored until clinical rejection without immunosuppression. In the MATCH group, flap survival did not significantly differ between the saline and hydrogen sulfide treatments (p = 0.483) but was reduced with the sodium iodide treatment (p = 0.007). In the MISMATCH group, survival was similar between the saline and hydrogen sulfide treatments (p = 0.483) but decreased with the sodium iodide treatment (p = 0.007). Rhabdomyolysis markers showed lower but non-significant levels in the experimental subgroups for both the MATCH and MISMATCH animals. This study provides insightful data for the field of antioxidant-based approaches in VCA and transplantation.

Subsidy-stress responses of ecosystem functions along experimental freshwater salinity gradients

Human activity is increasing salt concentrations in freshwaters worldwide, but effects of freshwater salinity gradients on biogeochemical cycling are less understood than in saline, brackish, or marine environments. Using controlled microcosm experiments, we characterized (1) short-term (one to five days) biogeochemical responses and (2) water column metabolism along a freshwater salinity gradient of multiple salt types. After one day, microcosms were oxic (4.48–7.40 mg O2 L−1) but became hypoxic (1.20–3.31 mg L−1) by day five. After one day in oxic conditions, microbial respiration in magnesium-, sodium-, and sea salt-based salinity treatments showed a subsidy-stress response, with respiration increasing by over 100% as salinity increased from 30 to 350–800 µS cm−1. Conversely, respiration consistently increased along a calcium-based salinity gradient, peaking at 1500 µS cm−1. By day five, an inverse subsidy-stress response was observed with elevated respiration at upper or lower ends of the gradient except for the magnesium treatment, which had the lowest respiration at the highest salinity. Calcium- and magnesium-based salinity treatments also caused considerable changes in phosphorus concentrations and C:P and N:P. In a separate experiment, microbial respiration and water column primary production also displayed subsidy-stress responses, but imbalances in effect sizes caused consistently declining net community production with increasing salinity. Collectively, our results establish that short-term exposure to different salt ion concentrations can enhance freshwater biogeochemical cycling at relatively low concentrations and alter resource stoichiometry. Furthermore, the nature of effects of freshwater salinization may also change with oxygen availability.

AquaCrop Model for Evaluation of the Water Productivity of Bread Wheat has Affected Modern Irrigation System and Water Salinity in Sandy Soil

AquaCrop is a very famous model in the world for evaluating the crop water productivity. FAO has developed the AquaCrop model as a leading and powerful scientific tool for simulating the response of different field crops to managing quantitative and qualitative irrigation. Two field experiments were carried out during winter seasons 2020/2021 and 2021/2022 at the Research and Production Station, National Research Centre, El-Nubaria, El-Behira Governorate, Egypt. to evaluate yield and yield component for two bread wheat varieties (Giza 171 and Misr 1) under sprinkler and drip irrigation systems with using four concentrations of water salinity irrigation. A split-plot design with three replications was used. Wheat were randomly distributed in the main plots while, while water salinity treatments occupied the sub-plots in both seasons. Results found that the simulated and observed values of water productivity were rather well in agreement for the salinity water levels treatments SWLT. This means that differences in canopy cover CC, yield and water productivity of bread wheat crop had significant effect on the calibration results. There is an inverse relationship between the water productivity and salinity water levels treatments and bread wheat varieties, while a direct correlation was found between water productivity and irrigation systems. It could be concluded that using sprinkler irrigation systems with S1 and S2 of SWLT and Misr 1 varitey, and drip irrigation system with S1 and S2 of SWLT and Misr 1 varitiey. the maximum of water productivity due to that there is a direct relationship between the study factors, the characteristics of the canopy cover growth and yield of the bread wheat plants, the more measured values will be simulated by the Aquacrop model.

Overexpression of the WRKY transcription factor gene NtWRKY65 enhances salt tolerance in tobacco (Nicotiana tabacum)

BackgroundSalt stress severely inhibits plant growth, and the WRKY family transcription factors play important roles in salt stress resistance. In this study, we aimed to characterize the role of tobacco (Nicotiana tabacum) NtWRKY65 transcription factor gene in salinity tolerance.ResultsThis study characterized the role of tobacco (Nicotiana tabacum) NtWRKY65 transcription factor gene in salinity tolerance using four NtWRKY65 overexpression lines. NtWRKY65 is localized to the nucleus, has transactivation activity, and is upregulated by NaCl treatment. Salinity treatment resulted in the overexpressing transgenic tobacco lines generating significantly longer roots, with larger leaf area, higher fresh weight, and greater chlorophyll content than those of wild type (WT) plants. Moreover, the overexpressing lines showed elevated antioxidant enzyme activity, reduced malondialdehyde content, and leaf electrolyte leakage. In addition, the Na+ content significantly decreased, and the K+/Na+ ratio was increased in the NtWRKY65 overexpression lines compared to those in the WT. These results suggest that NtWRKY65 overexpression enhances salinity tolerance in transgenic plants. RNA-Seq analysis of the NtWRKY65 overexpressing and WT plants revealed that NtWRKY65 might regulate the expression of genes involved in the salt stress response, including cell wall component metabolism, osmotic stress response, cellular oxidant detoxification, protein phosphorylation, and the auxin signaling pathway. These results were consistent with the morphological and physiological data. These findings indicate that NtWRKY65 overexpression confers enhanced salinity tolerance.ConclusionsOur results indicated that NtWRKY65 is a critical regulator of salinity tolerance in tobacco plants.

CAPN5 attenuates cigarette smoke extract-induced apoptosis and inflammation in BEAS-2B cells.

Apoptosis and chronic inflammation are the main phenotypes in chronic obstructive pulmonary disease (COPD) pathogenesis. Cigarette smoke exposure is the leading risk factor for COPD, which causes aberrant airway epithelial structure and function. As a non-classical calpain, the molecular function of calpain5 (CAPN5) in COPD remains unclear. This study investigated the role of CAPN5 in mediating cigarette smoke extract (CSE)-induced apoptosis and inflammation. Immunohistochemistry (IHC) and Western blotting (WB) were performed to detect the location and expression of CAPN5. In vitro, BEAS-2B cells were transfected with CAPN5 siRNA or CAPN5 plasmid, followed by phosphate-buffered saline (PBS) or cigarette smoke extract (CSE) treatment. The protein expression levels of CAPN5, NF-κB p65, p-p65, IκBα, p-IκBα and apoptosis proteins (BCL-2, BAX) were measured by WB. Flow cytometry (FCM) was performed to analyze the cell apoptosis index. CAPN5 was mainly expressed in the airway epithelium and significantly decreased in the COPD-smoker and emphysema-mouse groups. Silencing CAPN5 significantly decreased the protein expression of BCL-2, IκBα, and increased p-p65 and BAX protein expression. Additionally, an increased apoptosis index was detected after silencing CAPN5. Moreover, overexpression of CAPN5 partly inhibited IκBα degradation and p65 activation, and reduced CSE-induced inflammation and apoptosis. These combined results indicate that CAPN5 could protect against CSE-induced apoptosis and inflammation, which may provide a potential therapeutic target for smoking-related COPD.

Shenqi Chongcao Formula ameliorates inflammatory response in rats with pulmonary fibrosis by activating the ASS1/src/STAT3 signaling pathway

To observe the effect of Shenqi Chongcao (SQCC) Formula on the ASS1/src/STAT3 signaling pathway in a rat model of lung fibrosis and explore its therapeutic mechanism. A total of 120 male SD rats were divided equally into 5 groups, including a blank control group with saline treatment and 4 groups of rat models of idiopathic pulmonary fibrosis induced by intratracheal instillation of bleomycin. One day after modeling, the rat models were treated with daily gavage of 10 mL/kg saline, SQCC decoction (0.423 g/kg), pirfenidone (10 mL/kg), or intraperitoneal injection of arginine deiminase (ADI; 2.25 mg/kg, every 3 days) for 28 days. After the treatments, the lung tissues of the rats were collected for calculating the lung/body weight ratio, observing histopathology using HE and Masson staining, and analyzing the inflammatory cells in BALF using Giemsa staining. Serum chemokine ligand 2 (CCL2) and transforming growth factor-β1 (TGF-β1) levels were measured with ELISA. The protein expressions of src, p-srcTry529, STAT3, and p-STAT3Try705 and the mRNA expressions of ASS1, src and STAT3 in the lung tissues were detected using Western blotting and RT-qPCR. The neutrophil, macrophage and lymphocyte counts and serum levels of CCL2 and TGF-β1 were significantly lower in SQCC, pirfenidone and ADI treatment groups than in the model group at each time point of measurement (P < 0.05). P-srcTry529 and p-STAT3Try705 protein expression levels and ASS1, src, and STAT3 mRNA in the lung tissues were also significantly lower in the 3 treatment groups than in the model group (P < 0.05). SQCC Formula can alleviate lung fibrosis in rats possibly by activating the ASS1/src/STAT3 signaling pathway in the lung tissues.

Impact of microtrauma on the erythropoietic receptor of neuropathic Schwann cell-deficient mice

Charcot neuropathic arthropathy is a degenerative disease commencing with vasculopathic peripheral neuropathy and progresses with repeated micro-trauma, leading to instability, collapse, and severe foot and ankle disruption. Prior work suggests that erythropoietin’s (EPO) anti-apoptotic and anti-inflammatory effects may be beneficial in functional recovery of traumatized limbs. Employing a validated murine model, we investigated if diet induced obese (DIO) EPO knockout mice experience accelerated neurodegenerative changes and if EPO administration blunts the degree of neurodegeneration compared with saline treatment and those lacking EPO receptors on Schwann cells. Sixteen wild-type (C57BL6) and 16 Schwann cell-deficient (MPZ Cre) mice were given a high fat diet, completed a 3-day EPO protocol, and began a 10-week treadmill regimen. Serum blood glucose levels were collected. None of the groups had elevated median blood glucoses. Sensory, radiographic, and histopathologic analysis demonstrated no evidence of Charcot neuropathic arthropathy. The neurodegenerative mouse model previously achieved was thus not replicated. While these genetically modified, knockout mice serve as suitable models for studying the impact of EPO on nerve crush injuries, the current study was unable to achieve neurodegeneration, as suggested by the lack of neuropathy. Furthermore, our DIO EPO group failed to demonstrate fundamental prerequisite phenotypic changes necessary in the establishment of a neurodegenerative mouse model. This model supports prior research by demonstrating that in the absence of DIO, repetitive traumatic insult alone does not induce neurodegenerative changes. Without these changes, researchers cannot appreciate differences between groups or observe interventions’ impact, such as EPO, on the development of neurodegeneration.

Maximization of brackish water productivity for the sustainable production of striped catfish (Pangasianodon hypophthalmus) and grain sorghum (Sorghum bicolor (L.) Moench) cultivated under an integrated aquaculture–agriculture system

Freshwater scarcity, salinity, and poor soil fertility are the major challenges affecting both food and feed productions in arid and semi-arid regions of the world. Utilization of brackish water in the production of saline-tolerant fish and valuable field crops under an integrated system is promising in the maximization of yield per crop. The aim of this study, therefore, was to (1) assess the effect of saline aquaculture wastewater on the growth, yield, forage quality, and nutritive composition of sorghum seeds and (2) assess the effect of different water qualities on the survival, growth performance, and health status of Pangasianodon hypophthalmus. The experiment was conducted in a randomized completely block design of four salinity treatments with three replicates, i.e., control (freshwater mixed with inorganic fertilizers), 5000 ppm, 10,000 ppm, and 15,000 ppm. Our results indicated that although the control exhibited the highest growth (plant height, leaf number, internode number, leaf area, and soil–plant analysis development), grain, and forage yield, no significant differences were noted among the treatments. Likewise, no significant difference in the grain nutrient composition was noted among all the treatments. Assessment of the forage quality revealed improved crude protein content in the control compared to the saline treatments. However, no significant differences in the leaves and stalks fiber fractions were noted among all the treatments. Furthermore, rumen fermentation in terms of in vitro digestibility indicated no significant differences in the in vitro digestible dry matter, digestible organic matter, metabolic energy, net energy, microbial protein, short-chain fatty acids, and total dissolved nutrients among the treatments. However, rearing P. hypophthalmus in water salinities exceeding 10,000 ppm reduced the growth performance and health status of fish. Therefore, the integration of sorghum and P. hypophthalmus production in water salinities not exceeding 5000 ppm is a viable alternative to maximize brackish water productivity in freshwater-scarce regions.

Genome-wide identification and expression analyses of the Dicer-like, Argonaute and RNA-dependent RNA polymerase gene families in onion (Allium cepa L.)

ABSTRACT RNA silencing is a highly conserved gene expression regulatory mechanism participating in plant development and stress responses. Dicer-like protein (DCL), Argonaute protein (AGO) and RNA-dependent RNA polymerase (RDR) are directly involved in RNA silencing. So far, the characterisation and expression analysis of RDR, AGO and DCL proteins have not been performed in onion. A genome-wide analysis identified eight AcAGO, four AcDCL and four AcRDR putative genes in onion. Their cis-acting elements were investigated. The transcript levels of genes in different onion tissues and response to abiotic stress were studied. The results demonstrated that AcAGOs, AcDCLs and AcRDRs showed various tissue expression patterns. AcAGO2 and AcAGO4b were highly expressed in onion bulbs. AcDCLs and AcRDRs showed the highest transcript levels in inflorescence. AcAGO1, AcAGO5 and AcDCL1 were constitutively expressed in leaf and bulb during onion bulb expansion. AcRDR1 and AcRDR6 were upregulated during bulb expansion. AcAGOs were upregulated with salinity treatment, whereas AcDCLs and AcRDRs showed different expression patterns in abiotic stress response. Our results provided a basis for further clarification of RNA silencing-mediated environment stimulation response and development in onion.

Enhanced therapeutic effects of apoptotic cell‐conditioned mesenchymal stem cells in lupus‐prone MRL/lpr mice

AbstractBackgroundApoptotic cell‐conditioned mesenchymal stem cells (AC‐MSCs) exhibit stronger T cell suppressive ability via cyclooxygenase 2 (COX2)/prostaglandin E2 (PGE2); however, whether AC‐MSCs exhibit enhanced therapeutic effects on systemic lupus erythematosus (SLE) remains unknown.MethodsSplenocytes from MRL/MPJ‐Faslpr (MRL/lpr) mice were cocultured with AC‐MSCs, and the proportion of plasma cells was determined by flow cytometry. MSCs, AC‐MSCs, COX2 knockdown MSCs, and COX2 knockdown AC‐MSCs were infused into MRL/lpr mice (n = 10/group). Survival rates and lupus symptoms, including proteinuria, kidney injury, renal immune complex deposition, and autoantibody production, were assessed. Additionally, the number of plasma cells and serum levels of inflammatory cytokines were measured.ResultsThe AC‐MSCs significantly inhibited plasma cells via PGE2 after 24 h coculture in vitro, whereas MSCs did not. In the MRL/lpr mice, AC‐MSC treatment led to a significantly higher survival rate than phosphate‐buffered saline (PBS) treatment (90% vs. 50%, p &lt; 0.05). Moreover, AC‐MSC infusion decreased urine protein levels as early as 1 week after administration (0.89 ± 0.55 mg/mL vs. 1.59 ± 0.60 mg/mL, p &lt; 0.05, compared with PBS treatment). Administration of both MSCs and AC‐MSCs reduced renal immunoglobulin G and complement C3 deposition, whereas COX2 knockdown MSCs and COX2 knockdown AC‐MSCs did not. Serum anti‐dsDNA antibody levels in AC‐MSC‐treated mice significantly decreased (0.40 ± 0.25 vs. 0.99 ± 0.58, p &lt; 0.05), compared with PBS treatment, as well as the number of plasma cells in both the spleen ([2.14 ± 1.05] × 106 vs. [8.02 ± 4.01] × 106, p &lt; 0.01) and renal‐draining lymph nodes ([0.78 ± 0.68] × 106 vs. [2.49 ± 1.45] × 106, p &lt; 0.05). Additionally, AC‐MSCs inhibited the production of inflammatory cytokines, including interleukin‐21, tumor necrosis factor‐alpha, and monocyte chemoattractant protein‐1.ConclusionsAC‐MSCs enhanced the therapeutic effects in mice with lupus, which were partially mediated by COX2/PGE2. Therefore, AC preconditioning may be a new strategy for MSC transplantation in the treatment of SLE.

Mapping Single Nucleotide Polymorphism Markers Associated with the Pre-Flowering Morphological Performance of Fenugreek under Different Levels of Salt Stress

Salinity is a significant factor restricting plant growth and production. The effect of salinity stress on different growth parameters of 111 fenugreek genotypes was examined in an experiment with three salinity levels (0, 3000, 6000 mgL−1). A completely randomized block design with two replicated pots per treatment was used. Non-significant treatment effects were observed on fresh weight (FW); however, all traits showed significant genotype-by-treatment (GxT) interactions. This GxT was reflected in substantial SNP x environment interactions. Of 492 significant SNPs associated with the measured traits, 212 SNPs were linked to the correlated traits using an arbitrary threshold of three. Several SNPs were associated with FW and dry weight, measured under the same salinity treatment. The correlation between both traits was 0.98 under the three salinity treatments. In addition, 280 SNPs with conditional neutrality effects were mapped. The identified SNPs can be used in future marker-assisted breeding programs to select salt-tolerant genotypes. The results of this research shed light on the salt-tolerant properties of fenugreek.