Acid Stimulation Research Articles

Docosahexaenoic Acid Ameliorates the Toll-Like Receptor 22–Triggered Inflammation in Fish by Disrupting Lipid Raft Formation

BackgroundAlthough dietary DHA alleviates Toll-like receptor (TLR)–associated chronic inflammation in fish, the underlying mechanism is not well understood. ObjectivesThis study aimed to explore the role of Tlr22 in the innate immunity of large yellow croaker and investigate the anti-inflammatory effects of DHA on Tlr22-triggered inflammation. MethodsHead kidney–derived macrophages of croaker and HEK293T cells were or were not pretreated with 100 μM DHA for 10 h prior to polyinosinic-polycytidylic acid (poly I:C) stimulation. We executed qRT-PCR, immunoblotting, and lipidomic analysis to examine the impact of DHA on Tlr22-triggered inflammation and membrane lipid composition. In vivo, croakers (12.03 ± 0.05 g) were fed diets containing 0.2% [control (Ctrl)], 0.8%, and 1.6% DHA for 8 wk before injection with poly I:C. Inflammatory genes expression and rafts-related lipids and protein expression were measured in the head kidney. Data were analyzed by ANOVA or Student t test. ResultsThe activation of Tlr22 by poly I:C induced inflammation, and DHA diminished Tlr22-targeted inflammatory gene expression by 56–73% (P ≤ 0.05). DHA reduced membrane sphingomyelin (SM) and SFA-containing phosphatidylcholine (SFA-PC) contents, as well as lipid raft marker caveolin 1 amounts. Furthermore, lipid raft disruption suppressed Tlr22-induced Nf-κb and interferon h activation and p65 nuclear translocation. In vivo, expression of Tlr22 target inflammatory genes was 32–64% lower in the 1.6% DHA group than in the Ctrl group upon poly I:C injection (P ≤ 0.05). Also, the 1.6% DHA group showed a reduction in membrane SM and SFA-PC contents, accompanied by a decrease in caveolin 1 amounts, compared with the Ctrl group. ConclusionsThe activation of Tlr22 signaling depends on lipid rafts, and DHA ameliorates the Tlr22-triggered inflammation in both head kidney and head kidney–derived macrophages of croaker partially by altering membrane SMs and SFA-PCs that are required for lipid raft organization.

Hydrothermal Numerical Simulation of Injection Operations at United Downs, Cornwall, UK

The United Downs Deep Geothermal Project (UDDGP) is designed to utilize a presumably permeable steep dipping fault damage zone (constituting the hydrothermal reservoir in a very low permeability granitic host rock) for fluid circulation and heat extraction between an injection well at 2.2 km depth (UD−2) and a production well at 5 km depth (UD−1). Soft hydraulic stimulation was performed to increase the permeability of the reservoir. Numerical simulations are performed to analyze the hydraulic stimulation results and evaluate the increase in permeability of the reservoir. Experimental and field data are used to characterize the initial reservoir static model. The reservoir is highly fractured, and two distinct fracture networks constitute the equivalent porous matrix and fault zone, respectively. Based on experimental and field data, stochastic discrete fracture networks (DFN) are developed to mimic the reservoir permeability behavior. Due to the large number of fractures involved in the stochastic model, equivalent permeability fields are calculated to create a model which is computationally feasible. Hydraulic test and stimulation data from UD−1 are used to modify the equivalent permeability field based on the observed difference between the real fractured reservoir and the stochastic DFN model. Additional hydraulic test and stimulation data from UD−2 are used to validate this modified permeability. Results reveal that the equivalent permeability field model derived from observations made in UD−1 is a good representation of the actual overall reservoir permeability, and it is useful for future studies. The numerical simulation results show the amount of permeability changes due to the soft hydraulic stimulation operation. Based on the validated permeability field, different flow rate scenarios of the petrothermal doublet and their respective pressure evolution are examined. Higher flow rates have a strong impact on the pressure evolution. Simulations are performed in the acidized enhanced permeability region to make a connection between the ongoing laboratory works on the acid injection and field response to the possible acidizing stimulation.

Folic acid and zinc improve hyperuricemia by altering the gut microbiota of rats with high-purine diet-induced hyperuricemia.

A high-purine diet can cause hyperuricemia and destroy the microbial composition of the gut microbiota. Both folic acid and zinc significantly reduce uric acid levels and alleviate hyperuricemia. However, whether the underlying mechanisms are associated with the regulation of the gut microbiota remain unknown. To explore alterations of the gut microbiota related to folic acid and zinc treatment in rats with hyperuricemia in our study. A hyperuricemic rat model was established with a high-purine diet. The effects of folic acid and zinc on uric acid levels were evaluated. Alterations of the gut microbiota related to hyperuricemia and the treatments were evaluated by sequencing using the Illumina MiSeq system. The results demonstrated that uric acid levels dropped observably, and the activities of adenosine deaminase (ADA) and xanthine oxidase (XOD) were downregulated after folic acid or zinc intervention. 16S rRNA gene sequencing-based gut microbiota analysis revealed that folic acid and zinc enhanced the abundance of probiotic bacteria and reduced that of pathogenic bacteria, thus improving intestinal barrier function. PICRUST analysis indicated that folic acid and zinc restored gut microbiota metabolism. These findings indicate that folic acid and zinc ameliorate hyperuricemia by inhibiting uric acid biosynthesis and stimulating uric acid excretion by modulating the gut microbiota. Thus, folic acid and zinc may be new and safe therapeutic agents to improve hyperuricemia.

Physicochemical evaluation of the use of alcoholic micellar solutions containing nonylphenol and ethanol for the acidizing of carbonate matrices

Acid stimulation presents a great challenge when it comes to reducing the reaction rate between hydrochloric acid and carbonate rock, which makes it possible to increase the depth of the injected solutions. The surfactants are excellent candidates to reduce the reaction rate and make the rock water-wettable. However, high temperatures and the presence of salts as a product of the acidizing reaction can affect the performance of surfactants under reservoir conditions. In the present study, acidified alcoholic micellar solutions were obtained containing nonylphenol surfactant of 100, 40, and 11 ethoxylated bonds and ethanol as an amphi-solvent to assist the maintenance of the surfactant properties under adverse conditions. The solutions were characterized before and after the acidizing reactions (absence and presence of salt, respectively), evaluating the cloud point, surface tension, and kinematic viscosity. The experiments revealed that ethanol assisted in maintaining or improving the cloud point of the formulations, reaching above 70 °C. In addition, concerning nonylphenol 11EO formulations, the alcohol did not alter the effect of reducing the reaction rate and could partially substitute the surfactant in the active substance of the formulation.

The role of dietary plant and animal protein intakes on mitigating sarcopenia risk.

Purpose of reviewTo highlight contemporary findings comparing the digestibility of animal and plant proteins, their stimulatory effects on muscle protein synthesis, and associations with sarcopenia.Recent findingsAnimal proteins are more digestible than plant proteins, resulting in greater amino acid availability and stimulation of muscle protein synthesis. However, isolated plant proteins, plant protein blends, and modified plant proteins enriched with indispensable amino acids can elicit comparable digestion and absorption kinetics to animal proteins. More research is needed to determine whether these modified plant protein sources can effectively mitigate sarcopenia risk.SummaryBoth animal and plant protein foods can be incorporated into a healthful eating plan that limits risk of age-related diseases, such as sarcopenia. Humans eat food rather than isolated nutrients; as such, considering the context of the overall diet and its impact on health, instead of solely focusing on individual nutrients in isolation, is important.

Effect of Holstein genotype on ex-vivo cytokine response to lipopolysaccharide (LPS) and lipoteichoic acid (LTA) during the periparturient period

Effects of Holstein genotype on innate immune response were assessed with ex-vivo lipopolysaccharide (LPS) and lipoteichoic acid (LTA) stimulation of whole blood from unselected (UH, n = 10) and contemporary (CH, n = 11) Holsteins that differ in production by more than 4,500 kg/lactation. Blood was collected at −14, 7, 28, and 49 days in milk (DIM), mixed with a pathogen-associated molecular pattern (PAMP) molecule (0.01 or 1.0 µg LPS or 10 or 100 µg LTA per mL blood) and incubated (4 h, 37 °C). Plasma cytokines were quantified by ELISA, log10-transformed and analyzed by repeated measures with DIM as the repeated effect. Cytokine responses increased with PAMP dose and decreased as DIM increased. There was a genotype by LPS dose interaction for IL-1β as response to the low dose was greater in UH but did not differ between genotypes for the high dose. The IL-1β response was greater while the IL-6 response to LTA tended to be greater in UH than in CH cows. The more negative energy balance of CH cows did not impact genotype difference in cytokine responses. Results indicate selection since the mid-1960s has decreased ex-vivo, whole blood cytokine response of CH cows to LPS and to LTA.

Anisotropic chitosan/tunicate cellulose nanocrystals hydrogel with tunable interference color and acid-responsiveness

A robust chitosan/tunicate cellulose nanocrystals (TCNCs) anisotropic hydrogel with bright interference colors was fabricated via combining the prestretching orientation method and chemically-physically dual cross-linking. The oriented regenerated chitosan nanofibrous network enabled the TCNCs alignment by covalent interaction and hydrogen bonding. The stretching alignment endows the chitosan/TCNCs hydrogel with enhanced tensile strength, from 0.63 MPa (draw ratio 1.0) to 2.06 MPa (draw ratio 3.5). Moreover, the orientation of chitosan nanofibers led to birefringence appearance, which could be regulated with the TCNCs introduction or draw ratios. The hydrogel swelled completely in 2 min in pH = 3 solution and the interference color disappeared. The oriented chitosan/TCNCs hydrogels showed distinct color change under acid stimulation, which could be quantitatively measured or directly observed under crossed polarizers. This work demonstrated a strategy for fabricating the interference color regulatable hydrogels with acid-response property for sensors and environmental monitoring.

Human monocytes differentiate into tumor-associated macrophages upon SKOV3 cells coculture and/or lysophosphatidic acid stimulation

BackgroundSerous ovarian carcinoma is the most common type of ovarian carcinoma. Tumor-associated macrophages (TAMs) promote ovarian cancer progression. Most macrophages are generated by monocyte differentiation. Lysophosphatidic acid (LPA) levels are high in blood, tissues and ascites of patients with ovarian cancer. This study investigated whether human monocytes can directly differentiate into TAMs in the serous ovarian carcinoma microenvironment.MethodsHuman monocytes were isolated and purified from umbilical cord blood. A serous ovarian carcinoma-like microenvironment was generated by coculturing monocytes and SKOV3 cells in 0.4-μm-pore-size Transwell chambers. Additionally, the effect of LPA was assessed. The two cultured cell types and supernatants were evaluated.ResultsThe morphology and function of monocytes cocultured with SKOV3 cells and/or stimulated with LPA were significantly changed compared with those of non-stimulated monocytes. The CD14 + CD163 + and CD206 + phenotype indicated that stimulated cells were TAMs. The induced cells promoted SKOV3 cell proliferation and invasion, further proving that they were TAMs. The level of the cytokine interleukin-6R in the supernatant was significantly elevated in the treatment groups compared to the control monocyte group. Pathway enrichment analysis of ELISA results showed a strong influence of interleukin-6 family signaling, especially the JAK-STAT signaling pathway, further confirming the importance of IL-6R.ConclusionMonocytes can differentiate into TAMs under coculture with SKOV3 cells and/or LPA stimulation. The induced TAMs promote SKOV3 cell proliferation and invasion. The cytokine receptor IL-6sR and the JAK-STAT signaling pathway play an important role in the differentiation of monocytes into TAMs.

Effect mechanism of blistering moxibustion on visceral hypersensitivity of irritable bowel syndrome in mice based on 5-HT signal pathway

To observe the effect of blistering moxibustion on the expression levels of 5-hydroxytyptamine (5-HT) and its receptors of the colon tissue in the mice with visceral hypersensitivity of irritable bowel syndrome (IBS), so as to explore the effect mechanism of blistering moxibustion in treatment of IBS. Forty SPF-grade newborn Kunming mice were randomly divided into a normal group, a model group, an antagonist group and a blistering moxibustion group, 10 mice in each one. Before modeling, the injection with 0.2 mL parachlorophenylalanine (PCPA) was given on the lateral ventricle in the antagonist group. The endorectal glacial acetic acid stimulation combined with tail clipping was used to prepare the model of visceral hypersensitivity of IBS in the model group, the antagonist group and the blistering moxibustion group. After modeling, in the blistering moxibustion group, the intervention with blistering moxibustion was exerted at "Zhongwan" (CV 12), "Tianshu" (ST 25) and "Zusanli" (ST 36), once herbal irritant plaster at each acupoint, for 2 h each time, once a week, consecutively for 3 weeks. Abdominal withdrawal reflex (AWR) score and electromyographic (EMG) amplitude of abdominal muscles were adopted to evaluate the visceral hypersensitivity. HE staining was applied to observe the morphological changes in colon tissue, and immunohistochemistry was to determine the expression levels of 5-HT and its receptors. Compared with the normal group, EMG amplitude of abdominal muscles was increased under 20, 40 mm Hg (1 mm Hg=0.133 kPa) in the model group (P<0.05), AWR scores and EMG amplitude of abdominal muscles under 60, 80 mm Hg were all increased in the model group (P<0.05). In comparison with the model group, EMG amplitude of abdominal muscles was reduced under 20 mm Hg in the blistering moxibustion group (P<0.05), AWR scores were increased under 40 mm Hg in both the blistering moxibustion group and the antagonist group (P<0.05); AWR scores and EMG amplitude of abdominal muscles under 60, 80 mm Hg were all reduced in both the blistering moxibustion group and the antagonist group (P<0.05). Compared with the normal group, in the model group, the mucosa was slightly disturbed, while, the moderate inflammatory cells were visible in the submucosa. In comparison with the model group, the inherent glands of mucosa were regular in shape and a small number of inflammatory cells were visible in both the blistering moxibustion group and the antagonist group. In comparison with the normal group, the average positive staining area percentage (APSAP) of 5-HT and 5-HT3R of the colon tissue was increased, while, APSAP of 5-HT4R was reduced in the model group (P<0.05). Compared with the model group, APSAP of 5-HT and 5-HT3R was reduced in both the blistering moxibustion group and the antagonist group (P<0.05). Blistering moxibustion can relieve the visceral hypersensitivity of the mice with visceral hypersensitive IBS and the underlying mechanism is related to the regulation of the gut-brain axis mediated by 5-HT signaling pathway.

The Association Between Circulating Trans Fatty Acids and Thyroid Function Measures in U.S. Adults.

BackgroundThere has been controversial evidence regarding the effect of trans fatty acids (TFAs) on thyroid function in animal studies, and the epidemiological studies are lacking. We aimed to investigate the potential associations between circulating TFAs and thyroid function biomarkers in a U.S. adult population sample.MethodsWe performed a cross-sectional survey with 626 adults aged ≥20 years who participated in the National Health and Nutrition Examination Survey (NHANES) 2009–2010. Multivariable linear regression models were constructed to elucidate the relationships between circulating concentrations of TFAs (palmitelaidic acid, vaccenic acid, elaidic acid, linoelaidic acid and the sum of the four TFAs) and a panel of thyroid function measures.ResultsFor 626 adults, positive associations were found between palmitelaidic acid, elaidic acid and total thyroxine (TT4), between palmitelaidic acid and total triiodothyronine (TT3), and between linolelaidic acid and thyroid stimulating hormone (TSH), while linolelaidic acid was negatively associated with free thyroxine (FT4) (all P<0.05). Besides, the four TFAs and the sum TFAs were positively associated with free triiodothyronine (FT3). Vaccenic acid, elaidic acid, linoelaidic acid and the sum TFAs were positively associated with FT3/FT4, while the four TFAs and the sum TFAs were negatively associated with FT4/TT4 (all P<0.05). In stratified analysis, the associations between thyroid function measures and the ratios remained significant in female. For men, linolelaidic acid was negatively associated with FT4 and elaidic acid and the sum TFAs were positively associated with FT3. Furthermore, the associations between TFAs and FT3/FT4 remained significant.ConclusionOur findings revealed that TFAs exposure was associated with serum biomarkers of thyroid function. More researches are needed to evaluate the long-term health outcomes of these findings.

An Investigation of Insulinotropic Potential of Herbal Plants for Management of Diabetes

The Insulinotropic hormone glucagon-like peptide-1, which has been proposed as a modern treatment for management of diabetes, is metabolized extremely with the aid of dipeptidyl peptidase-IV. Inhibitors on dipeptidyl peptidase-IV enhance the level of glucagon-like peptide-1, as have elevated glucose tolerance and improved insulin secretion. Recently, incretin-based treatments have end up a beneficial tool to treat diabetic patients, and distinctive research have focused on the identification of glucagon-like peptide-1 receptor agonists, which includes these of herbal origin.&#x0D; Aims: This study was aimed to explorations the effect of methanolic extract of plants to inhibit dipeptidyl peptidase-IV and comparing its inhibitory activities with Diprotin, as a reference standard additionally, this study focuses to the influences of glucagon-like peptide-1secretions which accelerated glucose tolerance and provocation of insulin biosynthesis and secretion on STC-1 cell line.&#x0D; Study Design: In-vitro model.&#x0D; Place and Duration of Study: Department of pharmacology, Karnataka College of pharmacy, Bangalore, India, between Jan 2022 to April 2022.&#x0D; Methodology: The test drug undertaking of different medicinal plants extract of management over diabetes right here we were the methanolic extraction of three plants; “Aegle marmelos, Moringa oleifera, then Syzygium cumini” have been tested in-vitro for dipeptidyl peptidase-IV inhibitory activity and have an impact on of incretin system like glucagon-like peptide-1. An in-vitro assay to measure glucagon-like peptide-1release from cultured murine EEC’s under fatty acid stimulation. dipeptidyl peptidase-IV is involved in the inactivation over glucagon-like peptide-1, a potent Insulinotropic peptide.&#x0D; Results: The current study underlines up to expectation the extracts inhibits the dipeptidyl peptidase-IV and enhances the glucagon-like peptide-1for diabetes. Results established so the extracts on Aegle marmelos, Moringa oleifera, and Syzygium cumini had dipeptidyl peptidase-IV inhibitory activity on 42.57μg/mL, 42.38/mL, and then 41.48μg/mL respectively. Diprotin A confirmed an IC50 virtue on 29.83μg/mL, as is used as positive controls. Similarly, the study additionally demonstrates that on a cellular level of Aegle marmelos, Syzygium cumini potentially stimulate glucagon-like peptide-1secretion, however Moringa oleifera indicate decent rises into glucagon-like peptide-1secretion.&#x0D; Conclusion: The outcomes assure the inhibitory impact about plants regarding dipeptidyl peptidase-IV, glucagon-like peptide-1secretion, and the main in conformity with stand a novel, efficient and fair strategy for the management on diabetes.

Molecular characterization and expression analysis of TRIF, TRAF6, and TBK1 of golden pompano (Trachinotus ovatus)

Toll/IL-1R domain-containing adaptor-inducing IFN-β (TRIF), tumor necrosis factor receptor-associated factor 6 (TRAF6) and TANK-binding kinase 1 (TBK1) are critical signal transducers in toll-like receptors (TLRs) signaling pathway. In the present study, TRIF, TRAF6 and TBK1 were characterized from golden pompano (Trachinotus ovatus), named as TroTRIF, TroTRAF6 and TroTBK1, respectively. The full cDNA length of TroTRIF, TroTRAF6 and TroTBK1 was 2297 bp, 2293 bp, and 2482 bp, which respectively encoded 589, 573 and 723 amino acids. The deduced amino acids sequences of TroTRIF, TroTRAF6 and TroTBK1 contained conserved motifs, similar to their counterparts in other vertebrates. Phylogenetic tree analysis revealed that TroTRIF, TroTRAF6 and TroTBK1 were well clustered with their counterparts in other fish species. Quantitative Real-Time PCR (qPCR) analysis showed that TroTRIF, TroTBK1 and TroTRAF6 were detected in all examined tissues of healthy fish, but shared distinct transcript levels. Moreover, the expressions of TroTRIF, TroTBK1 and TroTRAF6 were generally induced by polyriboinosinic-polyribocytidylic acid (polyI:C), lipopolysaccharide (LPS), and Vibrio alginolyticus stimulation in vivo, indicating their critical roles in the immune defense of golden pompano against pathogen invasion. Our results provide valuable information for understanding the functions of these genes in golden pompano.

Quercetin ameliorated insulin resistance via regulating METTL3-mediated N6-methyladenosine modification of PRKD2 mRNA in skeletal muscle and C2C12 myocyte cell line

Background and aimsN6-Methyladenosine (m6A) modification is involved in many pathological processes, including insulin resistance (IR). Quercetin (Que), a bioactive compound with strong antioxidant activity, has potential therapeutic effects on IR-related metabolic diseases. The aim of this study is to investigate the roles of m6A and Que in hyperinsulinemia. Methods and resultsMale C57Bl/6 mice received a high-fat diet (HFD) for 8 weeks to establish an IR model. Que treatment reduced the body weight, blood glucose, plasma triglycerides (TG) and serum insulin, ameliorated IR, and decreased oxidative stress in HFD-fed mice. Cellular IR model was established in C2C12 cells by palmitic acid (PA) stimulation, and a noncytotoxic dose of Que was found to promote glucose uptake and inhibit oxidative stress. Moreover, methyltransferase-like 3 (METTL3) and serine–threonine kinase protein kinase D2 (PRKD2) was downregulated in skeletal muscle of HFD-fed mouse and in PA-induced C2C12 cells. The online bioinformatic tool SRAMP revealed that there were multiple m6A modification sites in the PRKD2 mRNA sequence. Downregulation of METTL3 enhanced PRKD2 expression by reducing m6A level and promoting mRNA stability in PRKD2 mRNA transcript. Que decreased m6A, METTL3, and phosphorylated insulin receptor substrate 1 (p-IRS1) levels, increased the protein expression of PRKD2, glucose transporter type 4 (GLUT4) and p-AKT, promoted glucose uptake, and reduced oxidative stress in PA-induced C2C12 cells. Moreover, METTL3 overexpression or PRKD2 silence reversed the inhibitory effects of Que on the levels of MDA and p-IRS1 and the promotive effects on glucose uptake, superoxide dismutase (SOD), GSH and GLUT4 and p-AKT levels. ConclusionQue promoted glucose uptake, repressed oxidative stress and improved IR through METTL3-mediated m6A of PRKD2 mRNA.

Decreased salivary α-amylase activity responding to citric acid stimulation in Myasthenia gravis with malnutrition.

ObjectivesMalnutrition, defined according to Nutritional risk screening (NRS 2002), is commonly observed in patients of Myasthenia gravis (MG), a neuromuscular disorder manifested by varied degrees of skeletal muscle weakness. Because biochemical composition of saliva changes in correspondence to alterations in nutritional status, we tested our hypothesis that a certain saliva component(s) might serve as a biomarker(s) for nutrition status of MG, particularly for those MG patients with high risk of malnutrition.Materials and methods60 MG patients and 60 subjects belonging to the healthy control group (HCG) were enrolled in this case-control study. The salivary α-amylase (sAA) activity, salivary flow rate (SFR), pH, total protein density (TPD), and the concentrations of chloride and calcium ions in MG group with or without malnutrition were measured before and after citric acid stimulation. Thereafter, the relationship between sAA activity and BMI was determined in MG and HCG.ResultsCompared with HCG, more patients with malnutrition, increased TPD and chloride and calcium concentrations but decreased pH value and SFR both before and after acid stimulation, as well as reduced sAA activity, pH and TPD responses to acid stimulation. MG with malnutrition showed decreased sAA activity and TPD responding to acid stimulation compared with those without malnutrition. Compared with normal BMI, sAA activity response to acid stimulation was reduced in low BMI. There was a significant strong positive correlation between the ratio of sAA activity and BMI in MG.ConclusionsSalivary biochemical characteristics are abnormally altered in MG with malnutrition. Altered sAA activity responding to acid stimulation was associated with malnutrition.Clinical relevanceDecreased sAA activity responding to acid stimulation can reflect malnutrition state and may be one potential screening marker for MG patients with high risk of malnutrition.

Lipid droplet dynamics in healthy and pyometra-affected canine endometrium

BackgroundAccumulation of lipid droplets (LDs) was recently observed in pyometra-affected uteri. As data about their nature and function are missing we intended to compare the localization, quality and quantity of LDs in canine healthy and pyometra-affected tissues and in an in vitro model.Methods and resultsWe characterized LDs in healthy and pyometra uterine tissue samples as well as in canine endometrial epithelial cells (CEECs) in vitro by means of histochemistry, immunohistochemistry, transmission electron microscopy, western blot, and RT-qPCR. Oil Red O (ORO) staining and quantification as well as p-phenylenediamine staining showed a higher number of LDs in epithelial cells of pyometra samples. Immunohistochemistry revealed that the amount of LDs coated by perilipin2 (PLIN2) protein was also higher in pyometra samples. Transmission electron microscopy showed an increase of LD size in surface and glandular epithelial cells of pyometra samples. In cell culture experiments with CEECs, supplementation with oleic acid alone or in combination with cholesterol lead to an increased LD accumulation. The expression of PLIN2 at protein and mRNA level was also higher upon oleic acid supplementation. Most LDs were double positive for ORO and PLIN2. However, ORO positive LDs lacking PLIN2 coating or LDs positive for PLIN2 but containing a lipid class not detectable by ORO staining were identified.ConclusionsWe found differences in the healthy and pyometra-affected endometrium with respect to LDs size. Moreover, several kinds of LDs seem to be present in the canine endometrium. In vitro studies with CEECs could show their responsiveness to external lipids. Since epithelial cells reacted only to oleic acid stimulation, we assume that the cyclic lipid accumulation in the canine endometrium is based mainly on triglycerides and might serve as energy provision for the developing early embryo. Further studies are necessary to verify the complex role of lipids in the healthy and pyometra-affected canine endometrium.

Effect of temperature on sandstone acidizing using non-Newtonian fluids

Rock formation is subjected to acid stimulation to increase oil reservoirs productivity. Acidizing of sandstone rocks is a common method to remove damage and recover the oil flow to the wellbore. A matrix acid stimulation will always require additives depending on mud acid (HF/HCl) components, which cause non-Newtonian fluid behavior. In this study, the 3-D mathematical model of sandstone matrix acidizing in radial flow has been presented by a modified two-parameter model to consider non-Newtonian fluids and temperature effects. To this end, the Carreau-Yasuda equation has been employed to predict dissolution patterns created by non-Newtonian reactive fluids in sandstone formations. Meanwhile, the energy equation has been solved in the porous media to estimate the temperature in each time step, which affects the dissolution patterns in matrix acidizing. Three main phenomena are involved in dissolution patterns: surface reaction, diffusion and convection. Acidizing simulation indicates the acid-rock reaction constant in the formation network is temperature sensitive, and increases exponentially by increasing the reservoir temperature. In this work, the effects of various heterogeneity magnitude and porosity in the porous media, injection rate, reaction rate constant, power-law index (n) and temperature are considered by the various presented dissolution patterns of sandstone acidizing. The results show that, to reach a certain penetration length, the pore volume (PV) of injected acid decreases sharply with reservoir temperature, but after the critical temperature, it decreases slowly. The model discretized the Darcy, mass and energy balance equations by finite difference method and pore evaluation by Karman-Cozney correlations, and then employed the Tri-Diagonal matrix algorithm (TDMA) method to achieve the solution of the model and analysis of dissolution patterns in various acidizing regimes and rheological properties. All mathematical estimations including Darcy correlations, mass and energy balance and pore evaluation have been solved in FORTRAN program by parallel processing to accelerate 3-D calculations.

Elevated Levels of Follicular Fatty Acids Induce Ovarian Inflammation via ERK1/2 and Inflammasome Activation in PCOS.

Polycystic ovary syndrome (PCOS) is accompanied by chronic inflammation and metabolic disorders. Whether metabolic abnormalities affect inflammation in PCOS or not, the underlying mechanism remains to be clarified. We aimed to investigate changes in fatty acids and their effects on inflammatory response in the follicular niche of PCOS patients. This study recruited 50 PCOS patients and 50 age-matched controls for follicular fluids and ovarian mural granulosa cells collection. The human ovarian granulosa cell line KGN was used for evaluating the effect of oleic acid (OA) stimulation. The levels of follicular fatty acids were measured by liquid chromatography-tandem mass spectrometry. The concentrations of inflammatory cytokines were detected by electrochemiluminescence and enzyme-linked immunosorbent assays. The regulation of inflammation-related genes was confirmed by quantitative polymerase chain reaction and Western blotting after OA stimuli. Three saturated fatty acids and 8 unsaturated fatty acids were significantly elevated in follicular fluids of PCOS patients compared to those in controls. The concentrations of follicular interleukin (IL)-6, IL-8, and mature IL-18 were significantly higher in the PCOS group and were positively correlated with the levels of fatty acids. Moreover, OA stimulation upregulated the transcription levels of IL-6 and IL-8 via extracellularly regulated kinase 1/2 signaling pathways in KGN cells. Furthermore, OA treatment induced reactive oxygen species production and inflammasome activation, which is manifested by enhanced caspase-1 activity and mature IL-18 protein level. Fatty acid metabolism was significantly altered in the follicular niche of PCOS patients. Elevated levels of fatty acids could induce ovarian inflammation both at the transcriptional level and in posttranslational processing.

Integrative Soil Application of Humic Acid and Foliar Plant Growth Stimulants Improves Soil Properties and Wheat Yield and Quality in Nutrient-Poor Sandy Soil of a Semiarid Region

Sandy soils (containing > 50% sand) are widely distributed worldwide and are characterized by their poor structure, low organic matter, weak hydraulic and nutritional properties, and low crop productivity. Using a 2-year pot experiment, in this study, we investigated the effects of humic acid (HA) as a soil amendment and study two plant growth stimulants (PGSs), zinc oxide nanoparticles (ZnONPs), and L-tryptophan (L-TRP), as a foliar application on wheat grown in nutrient-poor sandy soil. Three HA rates (0 (HA0), 0.2 (HA0.2), and 0.4 (HA0.4) g kg−1 soil) and five PGS levels [control, 50 mg l−1 (ZnONPs50), 100 mg l−1 (ZnONPs100), 0.25 mmol l−1 (L-TRP0.25), and 0.5 mmol l−1 (L-TRP0.5)] were used. The soil hydro-physico-chemical properties, morpho-physiological responses, yield, and quality were measured. HA addition amended the soil structure by allowing rapid macroaggregate formation, decreasing bulk density and pH, and increasing porosity and electrical conductivity, thereby improving soil hydraulic properties. HA0.2 and HA0.4 additions improved growth, yield components, and grain minerals, resulting in higher grain yield by 28.3–54.4%, grain protein by 10.2–13.4%, wet gluten by 18.2–23.3%, and dry gluten by 23.5–29.5%, respectively, than HA0. Foliar application of ZnONPs or L-TRP, especially at higher concentrations compared to the control, noticeably recorded the same positive results as HA treatments. The best results were achieved through the integration of HA0.4 + ZnONPs100 or L-TRP0.5 to the tested nutrient-poor sandy soil. The interactive application of HA0.4 + ZnONPs100 or L-TRP0.5 and the use of mineral fertilizer, which is considered a surplus point in permaculture, can be recommended for sustainable wheat production in nutrient-poor sandy soil.

Impaired surface marker expression in stimulated Epstein-Barr virus transformed lymphoblasts from Barth Syndrome patients

Primary B lymphocytes rapidly respond to lipopolysaccharide (LPS) and cytosine linked to a guanine by a phosphate bond deoxyribonucleic acid (CpG DNA) stimulation to promote adaptive immune function through increased surface marker expression. Here we examined expression of surface markers in LPS and CpG DNA stimulated Epstein-Barr virus transformed B lymphoblasts from control and BTHS patients with different mutations. The percentage of cluster of differentiation (CD) positive cells including CD38 + , CD138 + , CD80 + surface expression and programmed cell death protein 1 (PD1 +) surface expression was similar between control and BTHS lymphoblasts incubated plus or minus LPS. The percentage of CD24 + , CD38 + and CD138 + cells was similar between control and BTHS lymphoblasts incubated plus or minus CpG DNA. CD27 + surface marker expression was reduced in both BTHS lymphoblasts and controls incubated with CpG DNA and PD1 + surface marker expression was higher in BTHS cells compared to controls but was unaltered by CpG DNA treatment. Thus, Epstein-Barr virus transformed control and BTHS lymphoblasts fail to increase selected surface markers upon stimulation with LPS and exhibit variable surface marker expression upon stimulation with CpG DNA. Since B lymphocyte surface marker expression upon activation is involved in B cell proliferation and differentiation, cell–cell interaction and the adaptive immune response, we suggest that caution should be exercised when interpreting immunological data obtained from Epstein-Barr virus transformed BTHS cells. Based upon our observations in control cells, our conclusions may be more broadly applicable to other diseases which utilize transformed B lymphocytes for the study of immune biology.

Diurnal pattern of salivary alpha-amylase and cortisol under citric acid stimulation in young adults.

BackgroundSaliva composition has diurnal variations. Citric acid stimulation plays a major role in the change of salivary flow rate and salivary composition. However, diurnal variations and sex differences in salivary alpha-amylase (sAA), pH, salivary flow rate (SFR), and salivary cortisol before and after citric acid stimulation remain unclear.MethodsWe recruited 30 healthy volunteers, including 15 women (24.7 ± 1.0 years old) and 15 men (25.3 ± 1.3 years old). At four time points (T1, 7:00; T2, 10:00; T3, 16:00; and T4, 20:00), saliva was collected from healthy volunteers before and after citric acid stimulation; and sAA, pH, SFR and salivary cortisol were measured and compared between men and women.ResultsThere were circadian fluctuations in sAA activity, SFR, pH, and cortisol level both before and after citric acid stimulation, and the diurnal fluctuations of these indexes were not affected by citric acid stimulation. There were significant differences in salivary cortisol between men and women before and after acid stimulation in T1. Neither SFR nor pH showed sex-related differences before or after acid stimulation. The variation trend of sAA activity was contrary to that of cortisol, with a significant negative correlation.ConclusionsOur data suggest that sAA and cortisol showed diurnal fluctuation, and the variation characteristics of male and female under resting state and acid stimulation were basically the same. The variation trend of salivary alpha-amylase activity was opposite to that of cortisol, with significant negative correlation. Our findings may enable the selection of the correct sampling time for research and the selection of appropriate sampling strategies in studies investigating chronic psychosocial conditions.