Cyclic Adenosine Monophosphate Content Research Articles

A cAMP-biosensor-based assay for measuring plasma arginine–vasopressin levels

Arginine–vasopressin (AVP), a cyclic peptide hormone composed of nine amino acids, regulates water reabsorption by increasing intracellular cyclic adenosine monophosphate (cAMP) concentrations via the vasopressin V2 receptor (V2R). Plasma AVP is a valuable biomarker for the diagnosis of central diabetes insipidus (CDI) and is commonly measured using radioimmunoassay (RIA). However, RIA has several drawbacks, including a long hands-on time, complex procedures, and handling of radioisotopes with special equipment and facilities. In this study, we developed a bioassay to measure plasma AVP levels using HEK293 cells expressing an engineered V2R and a cAMP biosensor. To achieve high sensitivity, we screened V2R orthologs from 11 various mammalian species and found that the platypus V2R (pV2R) responded to AVP with approximately six-fold higher sensitivity than that observed by the human V2R. Furthermore, to reduce cross-reactivity with desmopressin (DDAVP), a V2R agonist used for CDI treatment, we introduced a previously described point mutation into pV2R, yielding an approximately 20-fold reduction of responsiveness to DDAVP while maintaining responsiveness to AVP. Finally, a comparison of plasma samples from 12 healthy individuals demonstrated a strong correlation (Pearson's correlation value: 0.90) between our bioassay and RIA. Overall, our assay offers a more rapid and convenient method for quantifying plasma AVP concentrations than existing techniques.

The Effect of Bacterial AHL on the Cyclic Adenosine Monophosphate Content in Plants According to High-Performance Liquid Chromatography.

Cyclic adenosine monophosphate (cAMP) is an important second messenger in cells, mediating various stimulation signals such as the growth and development of organisms and stress and participating in regulating various biological processes of cells. This article explores the quantitative determination of cAMP in plants using High-Performance Liquid Chromatography (HPLC) and applies this method to analyzing the changes in cAMP content during the process of plant response to the bacterial quorum sensing signal N-acyl homoserine lactone (AHL). Research has shown that the optimal detection conditions for HPLC are as follows: the chromatographic column is Venusil MP C18 (2), the mobile phase is methanol-water (0.1% trifluoroacetic acid) (v:v, 10:90), the detection wavelength is 259 nm, the column temperature is 35 °C, and the flow rate is 0.8 mL/min. The precision of the standard sample of this method is 98.21%, the precision of the sample is 98.87%, and the recovery rate is 101.067%. The optimal extraction conditions for cAMP in Arabidopsis are to use 15% methanol ultrasonic extraction for 10 min, followed by a 40 °C water bath for 4 h. Bacterial AHL signal processing can significantly stimulate an increase in cAMP levels in Arabidopsis leaves and roots. The establishment of HPLC detection methods for the cAMP content in plants is of great significance for in-depth research on the signal transduction mechanisms of plant-bacterial interactions.

FEATURES OF RELATIONS OF MELATONIN WITH THE STATE OF INTRACELLULAR REGULATORSOF THE FUNCTIONAL ACTIVITY OF WHOLE BLOOD MONONUCLEAR CELLS IN CORONARY HEART DISEASE

Introduction. Coronary heart disease (CHD), leading among the causes of death in adulthood and old age, is an urgent medical and social problem. The pathogenesis of most forms of coronary heart disease is based on stenosing atherosclerosis of the coronary arteries, which develops against the background of dyslipidemia and arterial hypertension and is accompanied by the activation of immunocompetent cells (ICCs) of the vascular wall with the development of a subclinical inflammatory reaction, as well as the production of pro-inflammatory factors such as interleukins, chemokines, growth factors and etc. In turn, ICC activity is determined by the state of their intracellular molecular cascades, which transmit signals into the cell and ensure its reactivity to various external stimuli, such as mitogens, cytokines, pathogen components, etcIt has been shown that the central nervous system plays an important role in the regulation of ICC activity due to the production of neurohumoral molecules, such as melatonin, endorphin, sero-tonin, etc., which ensure the coordination of immune responses and their control by the central nervous system. The aim of this study was to study the relationship between melatonin production and intracellular factors that regulate the pro-inflammatory activi-ty of whole blood mononuclear cells and their metabolism in patients with coronary artery disease. Material and methods. As part of the cohort study, 58 patients of both sexes with coronary artery disease aged 49 to 67 years and 20 practically healthy individuals of both sexes were examined. In accordance with the purpose of the study, the concentration of focal adhesion protein kinase (FAK), 5'AMP-activated protein kinase (AMPK), AKT1 protein kinase, signal transducers and transcription activators (STAT) was determined in nuclear cy-toplasmic lysates of whole blood mononuclear cells: STAT3, STAT5A and STAT6, c-Jun N-terminal protein kinase 1 and 2 isoforms (JNK), mitogen-activated protein kinase p38 (p38), extracellular growth kinase 1 and 2 isoforms (ERK), Janus kinase type 2 (JAK2), nuclear transcription factor NF -kB, caspase-1, cyclooxygenase-2 (COX-2), p70-S6K1 protein kinase, p53, p27, p21 proteins. In addition, the concentration of cyclic adenosine monophos-phate (cAMP) and cyclic guanosine monophosphate (cGMP) was determined in cell supernatants. Melatonin concentration was determined in blood se-rum. The material for the study was venous blood taken from the cubital vein in the morning from 6.00 to 6.15. Results. The analysis showed that in patients with coronary artery disease, in comparison with practically healthy individuals, in MNCs of whole blood, there was an increased level of protein kinases FAK, AKT, JNK, ERK, p70-S6K1, factor STAT6, protein p21, against which there was a decrease in the content of STAT3, STAT5A, JAK2, transcription factor NF-kB and caspase-1. These changes were accompanied by increased levels of cGMP and cAMP. Against this background, a decrease in the content of factors was revealed in the MNC. A high concentration of melatonin in patients with CAD was as-sociated with a decrease in the content of protein kinases AMPK, AKT, Jak2, ERK1, protein p21, caspase-1, and cAMP in MNCs, which was observed against the background of an increase in the level of protein p27 and nuclear factor NF-kB. The results of the correlation analysis indicate a different na-ture of the relationship between the level of melatonin and such factors as caspase-1, protein kinases ERK, JAK2, as well as the transcription factor NF-kB and p21 protein, depending on the characteristics of melatonin production in patients with coronary artery disease. Conclusions. In patients with coronary artery disease, melatonin exhibits a modulating effect on the energy balance of ICCs and their metabolism, helps limit pro-inflammatory activity by limiting the functional activity of MAPK/SAPK signaling pathways in MNCs.

Physiological and pathological mechanisms of oocyte meiosis.

Normal oogenesis is crucial to successful reproduction. During the human female fetal stage, primordial germ cells transform from mitosis to meiosis. After synapsis and recombination of homologous chromosomes, meiosis is arrested at the diplotene stage of prophase in meiosis I. The maintenance of oocyte meiotic arrest in the follicle is primarily attributed to high cytoplasmic concentrations of cyclic adenosine monophosphate. During the menstrual cycle, follicle-stimulating hormone and luteinizing hormone lead to the resumption of meiosis that occurs in certain oocytes and complete the ovulation process. Anything that disturbs oocyte meiosis may result in failure of oogenesis and seriously affect both the fertilization and embryonic development. The rapid development of the assisted reproduction technology, high-throughput sequencing technology, and molecular biology technology provide new ideas and means for human to understand molecular mechanism of meiosis and diagnosis and treatment of oocyte maturation defects. In this review, we mainly summarize the recent physiological and pathological mechanisms of oogenesis, involving homologous recombination, meiotic arrest and resumption, maternal mRNA degradation, post-translational regulation, zona pellucida assembly, and so on. We wish to take this opportunity to raise the awareness of researchers in related fields on oocyte meiosis, providing a theoretical basis for further research and disease treatments.

Expression of the umami taste receptor T1R1/T1R3 in porcine testis of: Function in regulating testosterone synthesis and autophagy in Leydig cells

Testosterone is a vital male hormone responsible for male sexual characteristics. The taste receptor family 1 subunit 3 (T1R3) regulates testosterone synthesis and autophagy in non-taste cells, and the links with the taste receptor family 1 subunit 1 (T1R1) for umami perception. However, little is known about these mechanisms. Thus, we aimed to determine the relationship between the umami taste receptor (T1R1/T1R3) and testosterone synthesis or autophagy in testicular Leydig cells of the Xiang pig. There was a certain proportion of spermatogenic tubular dysplasia in the Xiang pig at puberty, in which autophagy was enhanced, and the testosterone level was increased with a weak expression of T1R3. Silenced T1R3 decreased testosterone level and intracellular cyclic adenosine monophosphate (cAMP) content and inhibited the messenger RNA (mRNA) expression levels of testosterone synthesis enzyme genes [steroidogenic acute regulatory protein (StAR), hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1 (3β-HSD1), cytochrome P450 family 17 subfamily A member 1 (CYP17A1) and hydroxysteroid 17-beta dehydrogenase 3 (17β-HSD3)]. In addition, T1R3 increased the number of acidic autophagy bubbles and upregulated the expression levels of autophagy markers [Microtubule-associated protein 1 A/1B-light chain 3 (LC3) and Beclin-1] in testicular Leydig cells of the Xiang pig. Using an umami tasting agonist (10 mM L-glutamate for 6 h), the activation of T1R1/T1R3 enhanced the testosterone synthesis ability by increasing the intracellular cAMP level and upregulated the expression levels of StAR, 3β-HSD1, CYP17A1 and 17β-HSD3 in Leydig cells. Furthermore, the number of acidic autophagy bubbles decreased in the T1R1/T1R3-activated group with the downregulation of the expression levels of the autophagy markers, including LC3 and Beclin-1. These data suggest that the function of T1R1/T1R3 expressed in testicular Leydig cells of the Xiang pig is related to testosterone synthesis and autophagy.

Novel Melanocortin-3 and -4 Receptor Functional Variants in Asian Children With Severe Obesity.

Genetic variants in melanocortin 3 receptor (MC3R) and melanocortin 4 receptor (MC4R) genes are strongly associated with childhood obesity. This study aims to identify and functionally characterize MC3R and MC4R variants in an Asian cohort of children with severe early-onset obesity. Whole-exome sequencing was performed to screen for MC3R and MC4R coding variants in 488 Asian children with severe early-onset obesity (body mass index for age ≥97th percentile). Functionality of the identified variants were determined via measurement of intracellular cyclic adenosine monophosphate (cAMP) concentrations and luciferase activity. Four MC3R and 2 MC4R heterozygous nonsynonymous rare variants were detected. There were 3 novel variants: MC3R c.151G > C (p.Val51Leu), MC4R c.127C > A (p.Gln43Lys), and MC4R c.272T > G (p.Met91Arg), and 3 previously reported variants: MC3R c.127G > A (p.Glu43Lys), MC3R c.97G > A (p.Ala33Thr), and MC3R c.437T > A (p.Ile146Asn). Both MC3R c.127G > A (p.Glu43Lys) and MC4R c.272T > G (p.Met91Arg) variants demonstrated defective downstream cAMP signaling activity. The MC4R c.127C > A (p.Gln43Lys) variant showed reduced cAMP signaling activity at low substrate concentration but the signaling activity was restored at high substrate concentration. The MC3R c.151G > C (p.Val51Leu) variant did not show a significant reduction in cAMP signaling activity compared to wild-type (WT) MC3R. Coexpression studies of the WT and variant MC3R/MC4R showed that the heterozygous variants did not exhibit dominant negative effect. Our functional assays demonstrated that MC3R c.127G > A (p.Glu43Lys) and MC4R c.272T > G (p.Met91Arg) variants might predispose individuals to early-onset obesity, and further studies are needed to establish the causative effect of these variants in the pathogenesis of obesity.

Hetong decoction relieves loperamide-induced constipation in rats by regulating expression of aquaporins.

To investigate whether Hetong decoction (, HTT) alleviates constipation via regulating AQPs expression. Constipation in rats was induced by loperamide, and rats were randomly assigned into model (saline), HHT-low (95 g/kg), HTT-medium (190 g/kg), HTT-high (380 g/kg) and positive control (mosapride) groups. Then the defecation function, the concentration of serum arginine vasopressin (AVP) and cyclic adenosine monophosphate (cAMP), and the expression of AQP3 and AQP8 in colon tissues were assessed. NCM460 colon cells with AQP3 and AQP8 knockdown or overexpression were exposed to serum from rats that received low or high dose of HTT, followed by detection of AQP3 and AQP8 expression. The model group showed lower fecal weight and water content, weaker intestinal transit, higher serum concentration of AVP and cAMP, increased proximal and distal AQP8 expression, increased proximal but decreased distal AQP3 expression. However, these trends were reversed in both the HTT group (low, medium and high dose) and the positive control group. In NCM460 cells, HTT dose-dependently stabilized AQP3 and AQP8 expression under AQP3/8 plasmid interference or overexpression. HTT relieves constipation in rats through regulating AQP3 and AQP8 expression.

Pharmacogenomics in the Management of Pulmonary Arterial Hypertension: Current Perspectives.

Pulmonary arterial hypertension (PAH) is a rare disease with heterogeneous causes that can lead to right ventricular (RV) failure and death if left untreated. There are currently 10 medications representative of five unique pharmacologic classes that are approved for treatment. These have led to significant improvements in overall clinical outcome. However, substantial variability in dosing requirements and treatment response is evident, leading to suboptimal outcome for many patients. Furthermore, dosing is empiric and iterative and can lead to delays in meeting treatment goals and burdensome adverse effects. Pharmacogenomic (PGx) associations have been reported with certain PAH medications, such as treprostinil and bosentan, and can explain some of the variability in response. Relevant genes associated with treprostinil include CYP2C8, CYP2C9, CAMK2D, and PFAS. CYP2C8 and CYP2C9 are the genes encoding the major metabolizing liver enzymes for treprostinil, and reduced function variants (*2, *3) with CYP2C9 were associated with lower treatment persistence. Additionally, a higher CYP2C9 activity score was associated with a significantly less risk of treatment discontinuation. Other genes of interest that have been explored with treprostinil include CAMK2D, which is associated with right ventricular dysfunction and significantly higher dose requirements. Similarly, PFAS is associated with lower concentrations of cyclic adenosine monophosphate and significantly higher dose requirements. Genes of interest with the endothelin receptor antagonist (ERA) class include GNG2 and CYP2C9. A genetic variant in GNG2 (rs11157866) was linked to a significantly increased rate of clinical improvement with ERAs. The *2 variant with CYP2C9 (encoding for the major metabolizing enzyme for bosentan) was significantly associated with a higher risk for elevations in hepatic aminotransferases and liver injury. In summary, this article reviews the relevant pharmacogenes that have been associated to date with dosing and outcome among patients who received PAH medications.

Underlying mechnism of electroacupuncture for treating detrusor hyperreflexia after suprasacral spinal cord injury through the PACAP-cAMP signaling pathway: 基于PACAP-cAMP信号通路研究电针治疗骶上脊髓损伤后逼尿肌反射亢进型膀胱的效应机制

ObjectiveTo elucidate the underlying mechanism and effect of electroacupuncture (EA) on the neurogenic bladder following suprasacral spinal cord injury (SSCI). A rat model of detrusor hyperreflexia after SSCI was established to examine the urodynamics, detrusor muscle tissue morphology, the protein and mRNA expression levels of pituitary adenylate cyclase activating peptide (PACAP) and its receptor PAC1R, and cyclic adenosine monophosphate (cAMP) content in the detrusor muscle with a focus on the PACAP-cAMP signaling pathway. MethodA total of 72 female SD rats were randomized into control group and sham operation group (n=12 per group) by using a random number table. The remaining 48 rats were established into the model of detrusor hyperreflexia after SSCI. After successful modeling, these rats were randomly assigned to model, EA, and EA + PACAP6-38 groups (n=12 per group). The unsuccessful modeled rats were used for exploratory observation. For the rats in EA group, “Ciliao (BL32)” “Zhongji (CV3)”, and “Sanyinjiao (SP6)” were needled and stimulated by EA. The PACAP receptor antagonist PACAP6-38 was administered intraperitoneally in the EA + PACAP6-38 group before EA, and EA was applied for seven consecutive days. After treatment, the urodynamics of the rats were analyzed, and hematoxylin and eosin staining was used to examine rat bladder detrusor tissue morphology. The expressions of PACAP-38 and PAC1R were detected by immunohistochemistry and Western blot. The mRNA expression levels of PACAP-38 and PAC1R were examined by RT-qPCR, while cAMP content was detected by ELISA. Results(1) Compared with sham operation group, it was exhibited disarray in the transitional epithelium cells of the bladder in the modeled rats. The intercellular space was significantly widened, accompanied by inflammatory cell infiltration and noticeable tissue edema. Both the bladder initial pressure and leak point pressure of the rats were higher (P < 0.01), whereas the maximum cystometric capacity and bladder compliance were lower (P < 0.01). The protein and mRNA expression levels of PACAP-38 and PAC1R in the detrusor muscle, together with the cAMP content, were lower (P < 0.05). (2) Compared with the model rats, the EA group showed reduced inflammatory response in the detrusor muscle tissue, with decreased monocyte infiltration and less severe tissue edema. The bladder smooth muscle cells exhibited increased integrity, and there was decreased cellular tissue edema, inflammatory cell infiltration, and fibroplasia. The bladder initial pressure and leak point pressure were lower (P < 0.05), while the maximum cystometric capacity and bladder compliance were higher (P < 0.01). The protein and mRNA expression levels of PACAP-38 and PAC1R in the detrusor muscle, along with the cAMP content, were higher (P < 0.05). (3) Compared to the EA group, the EA + PACAP6-38 group showed a less organized arrangement of muscle fibers in the detrusor muscle tissue, larger intercellular space, monocyte infiltration, and considerable tissue edema. The changes in bladder initial pressure and leak point pressure were not significant (P > 0.05), while the maximum cystometric capacity and bladder compliance were lower (P < 0.05). The changes in the protein and mRNA expressions of PACAP-38 within the detrusor muscle were not significant (P > 0.05), whereas the protein and mRNA expressions of PAC1R were reduced (P < 0.05), and the cAMP content within the detrusor muscle was lower (P < 0.05). ConclusionEA can ameliorate the uninhibited contractile condition of the detrusor muscle in the bladder following SSCI. By mediating the PACAP-cAMP signaling pathway, it reduces the pathological damage to the detrusor muscle, thereby improving bladder function.

3-Hydroxybutyrate ameliorates insulin resistance by inhibiting PPARγ Ser273 phosphorylation in type 2 diabetic mice

3-Hydroxybutyrate (3HB) is a small ketone body molecule produced endogenously by the body in the liver. Previous studies have shown that 3HB can reduce blood glucose level in type 2 diabetic (T2D) patients. However, there is no systematic study and clear mechanism to evaluate and explain the hypoglycemic effect of 3HB. Here we demonstrate that 3HB reduces fasting blood glucose level, improves glucose tolerance, and ameliorates insulin resistance in type 2 diabetic mice through hydroxycarboxylic acid receptor 2 (HCAR2). Mechanistically, 3HB increases intracellular calcium ion (Ca2+) levels by activating HCAR2, thereby stimulating adenylate cyclase (AC) to increase cyclic adenosine monophosphate (cAMP) concentration, and then activating protein kinase A (PKA). Activated PKA inhibits Raf1 proto-oncogene serine/threonine-protein kinase (Raf1) activity, resulting in a decrease in extracellular signal-regulated kinases 1/2 (ERK1/2) activity and ultimately inhibiting peroxisome proliferator-activated receptor γ (PPARγ) Ser273 phosphorylation in adipocytes. Inhibition of PPARγ Ser273 phosphorylation by 3HB altered the expression of PPARγ regulated genes and reduced insulin resistance. Collectively, 3HB ameliorates insulin resistance in type 2 diabetic mice through a pathway of HCAR2/Ca2+/cAMP/PKA/Raf1/ERK1/2/PPARγ.

Silver Carp (Hypophthalmichthys molitrix) Scale Collagen Peptides-1 (SCPs1) Inhibit Melanogenesis through Downregulation of the cAMP-CREB Signaling Pathway.

The mechanism of silver carp scale collagen peptides (SCPs1) on melanogenesis and its mechanism of action were examined in mouse melanoma cells (B16). The cell viability and effects of SCPs1 on intracellular tyrosinase (TYR) activity and melanin, reactive oxygen species (ROS), glutathione (GSH) and cyclic adenosine monophosphate (cAMP) content were examined. The regulatory mechanism of SCPs1 on the cAMP response element-binding protein (CREB) signaling pathway was analyzed. The cell viability of the SCPs1 group was >80% (0.01-1 mg/mL) and the inhibitory rate of SCPs1 on B16 cell melanin increased in a dose-dependent manner. The highest inhibitory rate of SCPs1 on melanin content reaching 80.24%. SCPs1 significantly increased the GSH content and decreased the tyrosinase activity, as well as the content of ROS and cAMP. Western blot analysis showed that SCPs1 significantly inhibited melanocortin-1 receptor (MC1R) expression and CREB phosphorylation in the cAMP-CREB signaling pathway, leading to downregulation of microphthalmia-associated transcription factor (MITF) and the expression of TYR, TYR-related protein-1 (TRP-1) and TRP-2. SCPs1 also inhibited the expression of MC1R, MITF, TYR, TRP-1 and TRP-2 at the transcriptional level. Taken together, SCPs1 inhibited melanin synthesis through the downregulation of the cAMP-CREB signaling pathway. Fish-derived collagen peptides could potentially be applied in skin whitening products.

The Ras small GTPase RSR1 regulates cellulase production in Trichoderma reesei

BackgroundLignocellulose is the most abundant renewable resource in the world and has attracted widespread attention. It can be hydrolyzed into sugars with the help of cellulases and hemicellulases that are secreted by filamentous fungi. Several studies have revealed that the Ras small GTPase superfamily regulates important cellular physiological processes, including synthesis of metabolites, sporulation, and cell growth and differentiation. However, it remains unknown how and to what extent Ras small GTPases participate in cellulase production.ResultsIn this study, we found that the putative Ras small GTPase RSR1 negatively regulated the expression of cellulases and xylanases. Deletion of rsr1 (∆rsr1) significantly increased cellulase production and decreased the expression levels of ACY1-cAMP-protein kinase A (PKA) signaling pathway genes and the concentration of intracellular cyclic adenosine monophosphate (cAMP). Loss of acy1 based on ∆rsr1 (∆rsr1∆acy1) could further increase cellulase production and the expression levels of cellulase genes, while overexpression of acy1 based on ∆rsr1 (∆rsr1-OEacy1) significantly reduced cellulase production and transcriptional levels of cellulase genes. In addition, our results revealed that RSR1 negatively controlled cellulase production via the ACY1-cAMP-PKA pathway. Transcriptome analysis revealed significantly increased expression of three G-protein coupled receptors (GPCRs; tre62462, tre58767, and tre53238) and approximately two-fold higher expression of ACE3 and XYR1, which transcriptionally activated cellulases with the loss of rsr1. ∆rsr1∆ tre62462 exhibited a decrease in cellulase activity compared to ∆rsr1, while that of ∆rsr1∆tre58767 and ∆rsr1∆tre53238 showed a remarkable improvement compared to ∆rsr1. These findings revealed that GPCRs on the membrane may sense extracellular signals and transmit them to rsr1 and then to ACY1-cAMP-PKA, thereby negatively controlling the expression of the cellulase activators ACE3 and XYR1. These data indicate the crucial role of Ras small GTPases in regulating cellulase gene expression.ConclusionsHere, we demonstrate that some GPCRs and Ras small GTPases play key roles in the regulation of cellulase genes in Trichoderma reesei. Understanding the roles of these components in the regulation of cellulase gene transcription and the signaling processes in T. reesei can lay the groundwork for understanding and transforming other filamentous fungi.

Platelet zinc status regulates prostaglandin-induced signaling, altering thrombus formation

BackgroundApproximately 17.3% of the global population exhibits an element of zinc (Zn2+) deficiency. One symptom of Zn2+ deficiency is increased bleeding through impaired hemostasis. Platelets are crucial to hemostasis and are inhibited by endothelial-derived prostacyclin (prostaglandin I2 [PGI2]), which signals via adenylyl cyclase (AC) and cyclic adenosine monophosphate signaling. In other cell types, Zn2+ modulates cyclic adenosine monophosphate concentrations by changing AC and/or phosphodiesterase activity. ObjectivesTo investigate if Zn2+ can modulate platelet PGI2 signaling. MethodsPlatelet aggregation, spreading, and western blotting assays with Zn2+ chelators and cyclic nucleotide elevating agents were performed in washed platelets and platelet-rich plasma conditions. In vitro thrombus formation with various Zn2+ chelators and PGI2 was assessed in whole blood. ResultsIncubation of whole blood or washed platelets with Zn2+ chelators caused either embolization of preformed thrombi or reversal of platelet spreading, respectively. To understand this effect, we analyzed resting platelets and identified that incubation with Zn2+ chelators elevated pVASPser157, a marker of PGI2 signaling. In agreement that Zn2+ affects PGI2 signaling, addition of the AC inhibitor SQ22536 blocked Zn2+ chelation–induced platelet spreading reversal, while addition of Zn2+ blocked PGI2-mediated platelet reversal. Moreover, Zn2+ specifically blocked forskolin-mediated AC reversal of platelet spreading. Finally, PGI2 inhibition of platelet aggregation and in vitro thrombus formation was potentiated in the presence of low doses of Zn2+ chelators, increasing its effectiveness in inducing platelet inhibition. ConclusionZn2+ chelation potentiates platelet PGI2 signaling, elevating PGI2’s ability to prevent effective platelet activation, aggregation, and thrombus formation.

Effects of Shiwei Longdanhua formula on LPS induced airway mucus hypersecretion, cough hypersensitivity, oxidative stress and pulmonary inflammation

Shiwei Longdanhua Granule (SWLDH) is a classic Tibetan medicine (TM) ranking in the top 20 Chinese patent medicines in prescription rate to treat respiratory diseases like pneumonia, acute and chronic tracheobronchitis, acute exacerbation of COPD and bronchial asthma in solution of inflammation, cough and phlegm obstruction in clinical practice. However, its systematic pharmacological mechanisms have not been elucidated yet. Here, we studied the therapeutic efficacy of SWLDH in treatment of acute respiratory diseases in BALB/c mice by comprehensive analysis of airway inflammation, oxidative stress, mucus hypersecretion, cough hypersensitivities and indicators associated with the development of chronic diseases. Our results show that SWLDH might exhibit its inhibitory effects on pulmonary inflammation by interference with arachidonic acid (AA) metabolism pathways. Oxidative stress that highly related to the degree of tissue injury could be alleviated by enhancing the reductive activities of glutathione redox system, thioredoxin system and the catalytic activities of catalase and superoxide dismutase (SOD) after SWLDH treatment. In addition, SWLDH could significantly abrogate the mucus hypersecretion induced bronchiole obstruction by inactivate the globlet cells and decrease the secretion of gel-forming mucins (MUC5AC and MUC5B) under pathological condition, demonstrating its mucoactive potency. SWLDH also showed reversed effects on the release of neuropeptides that are responsible for airway sensory hypersensitivity. Simultaneously observed inhibition of calcium influx, reduction in in vivo biosynthesis of acetylcholine and the recovery of the content of cyclic adenosine monophosphate (cAMP) might collaboratively contribute to cause airway smooth muscle cells (ASMCs) relexation. These findings indicated that SWLDH might exhibited antitussive potency via suppression of the urge to cough and ASMCs contraction. Moreover, SWLDH might affect airway remodeling. We found SWLDH could retard the elevation of TGF-β1 and α-SMA, which are important indicators for hyperplasia and contraction during the progression of the chronic airway inflammatory diseases like COPD and asthma.

Autotetraploidization Alters Morphology, Photosynthesis, Cytological Characteristics and Fruit Quality in Sour Jujube (Ziziphus acidojujuba Cheng et Liu).

Artificially induced polyploidization is one of the most effective techniques for improving the biological properties and creating new cultivars of fruit trees. Up to now, systematic research on the autotetraploid of sour jujube (Ziziphus acidojujuba Cheng et Liu) has not been reported. 'Zhuguang' is the first released autotetraploid sour jujube induced with colchicine. The objective of this study was to compare the differences in the morphological, cytological characteristics, and fruit quality between diploid and autotetraploid. Compared with the original diploid, 'Zhuguang' showed dwarf phenotypes and decreased tree vigor. The sizes of the flowers, pollen, stomata, and leaves of 'Zhuguang' were larger. Perceptible darker green leaves were observed in 'Zhuguang' trees owing to increased chlorophyll contents, which led to higher photosynthesis efficiency and bigger fruit. The pollen activities and the contents of ascorbic acid, titratable acid, and soluble sugar in the autotetraploid were lower than those in diploids. However, the cyclic adenosine monophosphate content in autotetraploid fruit was significantly higher. The sugar/acid ratio of autotetraploid fruit was higher than that of diploid fruit, which made the autotetraploid fruit taste different and better. The results indicated that the autotetraploid we generated in sour jujube could greatly meet the goals of our multi-objective optimized breeding strategies for improving sour jujube, which includes tree dwarfing, increased photosynthesis efficiency, and better nutrient values and flavors as well as more bioactive compounds. Needless to say, the autotetraploid can also serve as material for generating valuable triploids or other types of polyploids and are also instrumental in studying the evolution of both sour jujube and Chinese jujube (Ziziphus jujuba Mill.).

Deucravacitinib and psoriasis: a sigh of relief

Introduction and pathophysiology Psoriasis is a chronic inflammatory skin condition linked to numerous other medical disorders, such as cardiometabolic syndrome, depression, and psoriatic arthritis1. Around 125 million individuals on the planet are affected by psoriasis. Genetic factors mostly cause psoriasis, while environmental variables can aggravate the condition2. Moderate-to-severe psoriasis is present in ~25% of affected patients3. Greater than 80% of cases of psoriasis fall within the most frequent subtype, plaque psoriasis. Erythematous scaly plaques, which typically develop on extensor surfaces like the knees and the elbows are the hallmark of this subtype. Gluteal fold, intertriginous areas, soles, palms, and nails may also be impacted. Furthermore, impacted regions are typically clearly defined and frequently symmetrical. A feed-forward mechanism of inflammation, notably the type 17 T-helper cell (TH17) pathway, leads to the development of plaque psoriasis2. Patients with more severe cases of psoriasis experience a stronger psychological impact as a result of the disease. Up to 20% of psoriasis sufferers are more prone to experience depression and have suicidal thoughts that progress to actual suicide4. Treatment options for psoriasis Primary treatment options for psoriasis include phototherapy and topical and systemic treatments which can be utilized singly or in combination. Phototherapy is initiated when the disease is extensive and resistant to other therapies. Two types of phototherapies are used: psoralen combined with ultraviolet A irradiation (PUVA) and UVB irradiation. However, they cost expensive and proper facilities are required. Despite toxicity-reducing strategies such as combination therapy, some individuals experience major adverse effects from traditional psoriasis medicines5. A phosphodiesterase 4 (PDE-4) inhibitor, Apremilast was approved by the Food and Drug Administration (FDA) in 2014 to treat plaque psoriasis6. Inhibition of PDE4 raises intracellular cyclic adenosine monophosphate (cAMP) concentrations. However, Apremilast’s precise therapeutic activity in people with psoriatic arthritis and psoriasis is not fully understood, nor are the mechanisms through which it works7. Apremilast over 16 weeks demonstrated a clinically significant improvement in patients suffering from plaque psoriasis, as determined by the PASI score, according to ESTEEM 1 and ESTEEM 2, two clinical trials8,9. Apremilast was the only oral drug available for plaque psoriasis until now. On September 9, 2022—FDA approved deucravacitinib, as a new and better oral treatment option for adults suffering from moderate to severe plaque psoriasis10,11. Sotyktu (deucravacitinib) Deucravacitinib is a tyrosine kinase 2 (TYK2) inhibitor that is selective and allosteric. It inhibits TYK2 by preferentially attaching to the pseudo kinase or unique regulatory domain instead of the enzyme’s active catalytic site12. The drug’s approval came after encouraging findings from the 2 clinical trials—POETYK PSO-1 and PSO-2, which found that Apremilast and placebo were less effective than deucravacitinib in treating 1684 adult patients with plaque psoriasis. In patients using deucravacitinib, oral ulcers, a rise in blood creatine phosphokinase, upper respiratory infections, pimples, and folliculitis were the most common side effects by the end of week 16 of these clinical studies. Deucravacitinib resulted in a rapid response and consistent trends in enhancements in many efficacy measurements as well as quality of life for those who have moderate to severe plaque psoriasis13. However, as per the manufacturer, Bristol Myers Squibb, combining Sotyktu (deucravacitinib) with other strong immunosuppressants is not advised. Conclusion For patients with psoriasis plaque who are not satisfied with conventional and topical therapies, the approval of deucravacitinib marks an exciting day, as it is the first oral medication authorized after a long time for plaque psoriasis. Given its track record of assisting patients in achieving smoother skin, as shown in the POETYK PSO clinical study, the drug deucravacitinib can change how psoriasis is managed and treated in a better way. Although the drug is contraindicated in pregnancy, information regarding the drug’s presence in human milk is lacking. Further research is required to comprehend better results in lactating women and widen the range as the drug has limitations in severe hepatic impairment10. Ethical approval Not applicable. Source of funding None. Author contributions MS.M. and F.A.: conceptualization. M.S.M., M.M.N.U., and S.T.A.: conducted the literature and drafting of the manuscript. M.A.S.: performed editing and supervision. All authors have read and agreed to the final version of the manuscript. Conflict of interest disclosure The authors declare that they have no financial conflict of interest with regard to the content of this report. Research registration unique identifying number (UIN) Not applicable. Guarantor All authors take responsibility for the work, access to data and decision to publish.

Regulatory role of myo-inositol in the ovarian development of female Chinese mitten crab (Eriocheir sinensis)

The ovarian development of female crab can directly affect the nutritional value, broodstock fertility and health status of offspring and are essential for the success of the crab industry. This study evaluates the influences of dietary myo-inositol (MI) on the ovarian development of female Chinese mitten crab (Eriocheir sinensis). Three diets containing different MI levels (0, 1600 and 3200 mg/kg) were fed to 144 crabs (initial weight: 72.38 ± 2.53 g) for 13 weeks. Crabs fed the diets with 1600, and 3200 mg/kg MI supplementation had significantly higher gonadosomatic index and yolk granule deposition in the ovary than those fed diets without MI supplementation. Crabs fed 1600 and 3200 mg/kg MI supplementation significantly enhanced vitellogenin synthesis in the hepatopancreas, vitellogenin contents in the hemolymph and vitellogenin receptor mRNA level in the ovary compared with those fed diets without MI supplementation. Simultaneously, cyclic adenosine monophosphate content and gene expression of retinoid X receptor, protein kinase A and ecdysteroid receptor in the hepatopancreas, and ecdysteroid content in hemolymph significantly increased when 1600 and 3200 mg/kg MI were supplemented in diets. Moreover, crabs fed 1600 and 3200 mg/kg MI supplementation had significantly higher lipid content in the ovary than those fed the diets without MI supplementation. The neutral lipid content in the hepatopancreas tended to decrease as the dietary MI level increased, while the polar lipid content in the hepatopancreas and the neutral and polar lipid contents in the ovary tended to increase. Dietary MI supplementation significantly enhanced the expression of genes related to lipid catabolism, lipid absorption and transport in the hepatopancreas, and gene expression related to lipid absorption in the ovary, suggesting that MI could promote the transport of lipids to the ovary from the hepatopancreas for ovarian development. This study indicates that dietary 1600 mg/kg MI supplementation is sufficient to promote the ovarian development of E. sinensis.

Inhibition of cyclooxygenase and EP3 receptor improved long term potentiation in a rat organotypic hippocampal model of repeated blast traumatic brain injury.

Blast-induced traumatic brain injury (bTBI) is a health concern in military service members who are exposed to multiple blasts throughout their training and deployment. Our group has previously reported decreased long term potentiation (LTP) following repeated bTBI in a rat organotypic hippocampal slice culture (OHSC) model. In this study, we investigated changes in inflammatory markers like cyclooxygenase (COX) and tested the efficacy of COX or prostaglandin EP3 receptor (EP3R) inhibitors in attenuating LTP deficits. Expression of COX-2 was increased 48h following repeated injury, whereas COX-1 expression was unchanged. EP3R expression was upregulated, and cyclic adenosine monophosphate (cAMP) concentration was decreased after repeated blast exposure. Post-traumatic LTP deficits improved after treatment with a COX-1 specific inhibitor, SC-560, a COX-2 specific inhibitor, rofecoxib, a pan-COX inhibitor, ibuprofen, or an EP3R inhibitor, L-798,106. Delayed treatment with ibuprofen and L-798,106 also prevented LTP deficits. These findings suggest that bTBI induced neuroinflammation may be responsible for some functional deficits that we have observed in injured OHSCs. Additionally, COX and EP3R inhibition may be viable therapeutic strategies to reduce neurophysiological deficits after repeated bTBI.

RNA interference reveals the escape response mechanism of Paramecium to mechanical stimulation.

In Paramecium, a mechanical stimulus applied to the posterior portion of the cell causes a transient increase in membrane permeability to potassium ions, transiently rendering the membrane in a hyperpolarized state. Hyperpolarization causes a transient increase in Cyclic adenosine monophosphate (cAMP) concentration in the cilia, resulting in a transient fast-forward swimming of the cell. Schultz and coworkers (1992) reported that a unique adenylate cyclase (AC)-coupled potassium channel is involved in the reaction underlying this response, which is known as the "escape response." However, the AC responsible for this reaction remains to be identified. Moreover, the molecular linkage between mechanoreception and AC activation has not been elucidated adequately. Currently, we can perform an efficient and simple gene-knockdown technique in Paramecium using RNA interference (RNAi). Paramecium is one of the several model organisms for which whole-genome sequences have been elucidated. The RNAi technique can be applied to whole genome sequences derived from the Paramecium database (ParameciumDB) to investigate the types of proteins that elicit specific biological responses and compare them with those of other model organisms. In this review, we describe the applications of the RNAi technique in elucidating the molecular mechanism underlying the escape response and identifying the AC involved in this reaction. The findings of this study highlight the advantages of the RNAi technique and ParameciumDB.

Growth kinetics, metabolomics changes, and antioxidant activity of probiotics in fermented highland barley-based yogurt

Fermenting yogurt with plant-based ingredients for nutritional and health benefits is an emerging trend. The growth kinetics, metabolomics changes, and antioxidant activity of probiotics in fermented highland barley-based yogurt were investigated in this study. Highland barley (HB) (Hordeum vulgare, Poaceae) enzyme hydrolyte was mixed with soy protein concentrate as ingredients for preparation of HB-based yogurt fermented by Lactobacillus bulgaricus VHProbi R03 and Streptococcus thermophilus VHProbi R08. After 4 h of fermentation, both strains grew to the stationary phase with the viable counts of 1.9 × 108 CFU/mL for each strain. The viable bacterial count in HB-based yogurt was approximately 9.3 × 107 CFU/mL after 21 days of storage at 4 °C. Compared with the control yogurt made using white sugar (as a substitute for HB enzyme hydrolyte) and soy protein concentrate, HB-based yogurt had higher concentrations of cyclic adenosine monophosphate, glycerophosphocholine, hypoxanthine and arginyl-proline, total phenolic compounds, and flavonoids, greater antioxidant activity, and higher sensory scores. In conclusion, HB-based yogurt is a potential novel food product that provides a good source of probiotics and antioxidants.