PA Treatment Research Articles

The multiple roles of macrophages in peritoneal adhesion

AbstractPeritoneal adhesion (PA) refers to the abnormal adhesion of the peritoneum either with the peritoneum itself or with tissues and organs that is caused by abdominopelvic surgery, abdominal infection or peritoneal inflammation. PA is associated with various clinical complications, such as abdominal pain and distension, intestinal obstruction, gastrointestinal disorders and female infertility, and adversely affects the quality of life of patients. Macrophages are essential for PA formation and can undergo polarization into classically activated macrophages (M1) and alternatively activated macrophages (M2), which are influenced by the peritoneal microenvironment. By releasing proinflammatory cytokines and reactive oxygen species, M1 macrophages promote peritoneal inflammatory reactions and the resultant formation of adhesion. In contrast, M2 macrophages secrete anti‐inflammatory cytokines and growth factors to inhibit PA formation and to promote repair and healing of peritoneal tissues, and thereby play a significant anti‐inflammatory role. This review comprehensively explores the function and mechanism of macrophages and their subtypes in PA formation to gain insight into the prevention and treatment of PA based on the modulation of macrophages.

Pituitary apoplexy is associated with concurrent or subsequent diagnosis of human immunodeficiency virus

AbstractObjectivePituitary apoplexy (PA) is a rare condition caused by hemorrhage or infarction of the pituitary gland with associated risk of cranial neuropathies, endocrinopathies, and even death. Human immunodeficiency virus (HIV) has been shown to have some association with pituitary apoplexy, however, large‐scale studies have not been performed.MethodsA retrospective cohort study of subjects greater than 18 years of age was performed using the national TriNetX database. The general population was first compared with those undergoing treatment for PA to compare general demographics and incidence of HIV. The PA cohort was then substratified by HIV status and propensity matched by age and sex to evaluate differences in visual outcomes and endocrine metrics. Using the diagnosis of pituitary apoplexy as an index event, the rate of HIV diagnosis at the time of presentation or in the three months following treatment was calculated. Two‐tailed, unpaired t‐tests were performed.ResultsA total 2066 patients were identified (902 HIV, 1158 controls). Those with HIV showed an increased rate of pituitary apoplexy (odds ratio [OR]: 19.8) versus healthy adults and were more likely to be younger and male. Among patients treated for pituitary apoplexy, patients with HIV were more likely to have increased thyroid stimulating hormone, reduced T4, and increased prolactin compared with healthy controls. There were no significant differences in visual outcomes between HIV positive and control patients. Importantly, 74% of patients did not carry a diagnosis of HIV on presentation but were diagnosed at the time of or in the three months following treatment for pituitary apoplexy.ConclusionsDiagnosis and treatment of pituitary apoplexy shows a high rate of concurrent or subsequent diagnosis of HIV. Screening for HIV should be considered in patients undergoing treatment for pituitary apoplexy.

PENGARUH WAKTU PENYERBUKAN TERHADAP KUALITAS HASIL TANAMAN MELON (Cucumis melo L) DENGAN PERKAWINAN SILANG TANAMAN SEMANGKA (Citrullus vulgaris L)

Decreased production of melon (Cucumis melo L) which is due to constraints in fruit quality decreases due to the ability of fruit formation is naturally low due to a failure in the process of pollination. The success of pollination it self is influenced by the maturity of the male flowers and female flowers itself. Therefore we need a suitable time to perform pollination to see the stigma receptivity and pollen viability at the same level. To overcome these problems need to develop crops melon (Cucumis melo L) through the technique of cultivation is called plant breeding to obtain the production of melon (Cucumis melo L) qualified by determining the time of pollination and crossed with a watermelon (Citrullus vulgaris L). This research is experimental research design using a randomized block design (RAK) 1 factor with 5 treatment combinations and each treatment using 10 replications. The treatment factors are as follows: PA = at 05:00 to 06:00, PB = at 07:00 to 08:00, PC = at 09:00 to 10:00, PD = at 11 : 00 to 12:00 and PE = at 13:00 to 14:00. The data were collected and presented in tables and statistical analysis of test One-Way ANOVA (Analysis Of Variance). If there is a real difference followed by testing of LSD (Least Significant Difference) (p <0.05). Statistical analysis One-Way ANOVA test and LSD show that there is the influence of pollination time on the fruit weight, average fruit diameter and weight of seed resulting from cross-breeding melon (Cucumis melo L) and watermelon (Citrullus vulgaris L). Data show that the PA treatment is the best treatment compared with other treatments. This happens Because the quality of pollen in the morning are of better quality compared to during the day so that pollen powderd in the morning better able to fertilize the pistil to the maximum.

Protective effect of SERCA2a-SUMOylation by SUMO-1 on diabetes-induced atherosclerosis and aortic vascular injury.

Diabetes is a major risk factor for cardiovascular disease. However, the exact mechanism by which diabetes contributes to vascular damage is not fully understood. The aim of this study was to investigate the role of SUMO-1 mediated SERCA2a SUMOylation in the development of atherosclerotic vascular injury associated with diabetes mellitus.ApoE-/- mice were treated with streptozotocin (STZ) injection combined with high-fat feeding to simulate diabetic atherosclerosis and vascular injury. Human aortic vascular smooth muscle cells (HAVSMCs) were treated with high glucose (HG, 33.3 mM) and palmitic acid (PA, 200 µM) for 24h to mimic a model of diabetes-induced vascular injury in vitro. Aortic vascular function, phenotypic conversion, migration, proliferation, intracellular Ca2+ concentration, the levels of small ubiquitin-like modifier type 1 (SUMO1), SERCA2a and SUMOylated SERCA2a were detected.Diabetes-induced atherosclerotic mice presented obvious atherosclerotic plaques and vascular injury, companied by significantly lower levels of SUMO1 and SERCA2a in aorta. HG and PA treatment in HAVSMCs reduced the expressions of SUMO1, SERCA2a and SUMOylated SERCA2a, facilitated the HAVSMCs phenotypic transformation, proliferation and migration, attenuated the Ca2+ transport, and increased the resting intracellular Ca2+ concentration. We also confirmed that SUMO1 directly bound to SERCA2a in HAVSMCs. Overexpression of SUMO1 restored the function and phenotypic contractile ability of HAVSMCs by upregulating SERCA2a SUMOylation, thereby alleviating HG and PA-induced vascular injury.These observations suggest an essential role of SUMO1 to protect diabetes-induced atherosclerosis and aortic vascular injury by the regulation of SERCA2a-SUMOylation and calcium homeostasis.

(066) Targets of SHH Signaling, BMP4 and GREM1, are Downstream Regulators of Smooth Muscle in the Penis

Abstract Introduction A key aspect of erectile dysfunction (ED) development is loss of innervation to the penis. The cavernous nerve (CN) is frequently damaged in prostatectomy and diabetic patients with ED, initiating changes in penile morphology including an acute and intense phase of apoptosis in penile smooth muscle followed by increasing collagen and fibrosis. This remodeling process alters penile architecture so that there is less smooth muscle in the corpora cavernosa, and what remains is less compliant/able to relax in response to normal neurotransmitter signaling, and ED results. Thus, novel therapies are needed which target the underlying remodeling process. We have shown previously that Sonic hedgehog (SHH) is a critical regulator of penile smooth muscle, and that SHH protein treatment suppresses penile remodeling after CN injury through an unknown mechanism. Objective We examine if part of the mechanism of how SHH preserves smooth muscle after CN injury involves bone morphogenetic protein 4 (BMP4) and GREMLIN (GREM1). Methods Primary cultures of smooth muscle cells were established from patients who had prostatectomy, diabetes, hypertension and Peyronie’s (control, n=18). Cultures were characterized by immunohistochemical analysis (IHC) for ACTA2, CD31, P4HB and nNOS. Smooth muscle cell growth was quantified in response to BMP4 and GREM1. Adult Sprague Dawley rats that were uninjured or underwent CN crush injury, were treated with BMP4, GREM1 or MSA (control) proteins via Affi-Gel beads (n=16) or peptide amphiphile (PA, n=26) for 3 and 14 days, and trichrome stain was performed. Sprague Dawley rats underwent CN injury (n=9) and SHH PA treatment for 1, 2 and 4 days (n=9) and western was performed assaying for BMP4 and GREM1 proteins in comparison to sham (n=3) controls. Adult Sprague Dawley rats were treated with 5E1 SHH inhibitor (n=6) or IgG (control, n=6) for 2 and 4 days, and BMP4 and GREM1 localization were examined by IHC. Statistics were performed by ANOVA with Scheffe’s posthoc test. Results Patient primary smooth muscle cultures were established. BMP-4 increased patient smooth muscle cell growth and GREM1 decreased growth. In rats BMP4 treatment via Aff-Gel beads and PA increased smooth muscle growth at 3 and 14 days of treatment. GREM1 treatment caused collagen and smooth muscle growth at 3 days, which switched to primarily collagen at 14 days. CN injury increased BMP4 and GREM1. SHH PA altered western band size suggesting alternative cleavage and range of signaling. SHH inhibition in rats changed BMP4 and GREM1 localization. Conclusions BMP4 treatment increases smooth muscle in patient cultures and in our rat model. BMP4 and GREM1 mediate a switch from smooth muscle to collagen induction. SHH treatment alters BMP4 and GREM1 localization and range of signaling, which can affect penile morphology. Whether GREM1 directly effects collagen induction or acts strictly through inhibition of BMP4 is unknown and requires further study. Part of the mechanism of how SHH regulates corpora cavernosal smooth muscle, involves BMP4 and GREM1. Understanding how SHH PA impacts penile morphology after CN injury will aid development of ED therapies. Disclosure No.

Nitrogen addition regulates the effects of variation in precipitation patterns on plant biomass formation and allocation in a Leymus chinensis grassland of northeast China.

Global warming is predicted to change precipitation amount and reduce precipitation frequency, which may alter grassland primary productivity and biomass allocation, especially when interact with other global change factors, such as nitrogen deposition. The interactive effects of changes in precipitation amount and nitrogen addition on productivity and biomass allocation are extensively studied; however, how these effects may be regulated by the predicted reduction in precipitation frequency remain largely unknown. Using a mesocosm experiment, we investigated responses of primary productivity and biomass allocation to the manipulated changes in precipitation amount (PA: 150mm, 300mm, 450mm), precipitation frequency (PF: medium and low), and nitrogen addition (NA: 0 and 10 g N m-2 yr-1) in a Leymus chinensis grassland. We detected significant effects of the PA, PF and NA treatments on both aboveground biomass (AGB) and belowground biomass (BGB); but the interactive effects were only significant between the PA and NA on AGB. Both AGB and BGB increased with an increment in precipitation amount and nitrogen addition; the reduction in PF decreased AGB, but increased BGB. The reduced PF treatment induced an enhancement in the variation of soil moisture, which subsequently affected photosynthesis and biomass formation. Overall, there were mismatches in the above- and belowground biomass responses to changes in precipitation regime. Our results suggest the predicted changes in precipitation regime, including precipitation amount and frequency, is likely to alter primary productivity and biomass allocation, especially when interact with nitrogen deposition. Therefore, predicting the influence of global changes on grassland structure and functions requires the consideration of interactions among multiple global change factors.

A single-center experience of non-bioartificial DFAPP support systems among Chinese patients with hyperlipidemic moderate/severe acute pancreatitis

To assess the clinical efficacy of Double Filtration Plasmapheresis (DFAPP), a novel blood purification method, in treating hyperlipidemic moderate/severe pancreatitis (HL-M/SAP). A total of 68 HL-M/SAP patients were enrolled in this study. The observation group, comprising 34 patients, received DFAPP treatment, while the control group underwent CVVH + PA treatment. We compared the efficacy changes between the two groups post-treatment. Patients treated with DFAPP showed significant improvements in clinical outcomes. After 72 h of DFAPP treatment, HL-M/SAP patients exhibited notably lower multiple organ failure scores and a reduced mortality rate compared to those in the CVVH + PA group. Triglyceride levels in HL-M/SAP patients treated with DFAPP for 48 h averaged 3.75 ± 1.95, significantly lower than the 9.57 ± 3.84 levels in the CVVH + PA group (P < 0.05). Moreover, CRP levels decreased markedly, IL-17 levels diminished, IL-10 levels increased, and the decline in IL-35 levels was significantly less pronounced compared to the CVVH + PA group. The recurrence rate of pancreatitis was also significantly lower after 6 months. The early implementation of DFAPP in HL-M/SAP patients effectively reduces triglyceride levels, suppresses pro-inflammatory factors, enhances anti-inflammatory factors, and mitigates cytokine storm-induced sepsis damage. Consequently, this leads to a decrease in the incidence of multiple organ failure, improved patient survival rates, and a reduce the recurrence rate of lipogenic pancreatitis.Trial registration: Chinese Clinical Trial Registry, ChiCTR2300076066.

Pentadecanoic acid (C15:0, PA) induces mild maternal glucose intolerance and promotes the growth of the offspring partly through up-regulating liver PPARα and MAPK signaling pathways.

Gestational diabetes mellitus (GDM) is one of the most common metabolic disturbances during pregnancy, which poses a serious threat to both maternal and offspring health. Pentadecanoic acid (C15:0, PA) is one of the most common odd-chain saturated fatty acids (OCS-FAs). However, its safety and nutritional value are yet to be verified. Herein, we provide a systematic assessment of the effects of PA on maternal and progeny health and insulin sensitivity for the first time. Our results showed that consumption of 1% PA during pregnancy could increase the contents of PA and heptadecanoic acid (C17:0) in maternal plasma, fetal tissue and offspring plasma, but it had no effect on embryonic development. During pregnancy, PA treatment caused mild insulin resistance, while it had little effect on the maternal body composition. During lactation, PA treatment caused mild insulin resistance and oxidative stress. Maternal body fat deposition was also reduced, but the growth rate of the offspring was faster. It is worth noting that PA treatment decreased plasma and liver TG content and increased the antioxidant capacity of the offspring. The effect of PA on the transcription and expression genes in the liver of pregnant mice was investigated using RNA-seq. PPARα and MAPK signaling pathways, both closely related to lipolysis, inflammation, oxidative stress, and insulin resistance were significantly increased. The expression of c-JUN, ERK, JNK and P65 proteins was also significantly up-regulated. In conclusion, our results suggest that 1% PA can induce a mild decrease in the maternal glucose tolerance and lipolysis mainly by activated MAPK and PPARα signaling. Moreover, low concentrations of PA may be an effective nutrient to alleviate the oxidative stress and reduce blood lipid levels of offspring.

Milk production responses of dairy cows to fatty acid supplements with different ratios of palmitic and oleic acids in low- and high-fat basal diets

We evaluated the effects of fatty acid (FA) supplements with different ratios of palmitic acid (C16:0) and oleic acid (cis-9 C18:1) in low- and high-FA basal diets on production responses of lactating dairy cows. Thirty-six multiparous Holstein cows (50.2 ± 5.8 kg/d of milk; 160 ± 36 d in milk) were used in a split-plot Latin square design balanced for carryover effects. Cows were blocked by milk yield and allocated to a main plot receiving either a low-FA (LF; 1.93% FA content) basal diet (n = 18) containing cottonseed meal and cottonseed hulls or a high-FA (HF; 3.15% FA content) basal diet (n = 18) containing whole cottonseed. Within each plot, a 3 × 3 Latin square arrangement of treatments was used in 3 consecutive 21-d periods. Treatments were (1) control (CON; no FA supplementation), (2) FA supplement containing 80% C16:0 + 10% C18:1 (PA), and (3) FA supplement containing 60% C16:0 + 30% cis-9 C18:1 (PA+OA). The FA supplements were fed at 1.5% of dry matter and replaced soyhulls in CON. Preplanned contrasts were (1) overall effect of FA supplementation {CON vs. the average of the FA treatments [1/2 (PA + PA+OA)]}, and (2) the effect of the PA treatment versus the PA+OA treatment (PA vs. PA+OA). Treatment by basal diet interactions were observed for yields of milk and lactose, where FA treatments increased yields of milk and milk lactose in LF but not in HF. Basal diet had no effect on dry matter intake (DMI) or milk yield. Compared with LF, HF increased milk fat yield and 3.5% fat-corrected milk (FCM) and tended to increase milk fat content and energy-corrected milk (ECM) yield. The FA treatments decreased DMI but increased the yields of milk fat, 3.5% FCM, and ECM, compared with CON, due to increases in mixed and preformed milk FA yields. The PA+OA treatment decreased DMI and milk protein yield compared with PA. In conclusion, a high-fat basal diet increased milk fat production, and the addition of FA supplements to a low-fat basal diet increased milk lactose yield and tended to increase milk yield. Additionally, regardless of basal diet fat level, FA supplements increased production responses compared with the non-FA-supplemented control diet.

Face and Predictive Validity of MI-RAT (Montreal Induction of Rat Arthritis Testing), a Surgical Model of Osteoarthritis Pain in Rodents Combined with Calibrated Exercise.

Validating animal pain models is crucial to enhancing translational research and response to pharmacological treatment. This study investigated the effects of a calibrated slight exercise protocol alone or combined with multimodal analgesia on sensory sensitivity, neuroproteomics, and joint structural components in the MI-RAT model. Joint instability was induced surgically on day (D) 0 in female rats (N = 48) distributed into sedentary-placebo, exercise-placebo, sedentary-positive analgesic (PA), and exercise-PA groups. Daily analgesic treatment (D3-D56) included pregabalin and carprofen. Quantitative sensory testing was achieved temporally (D-1, D7, D21, D56), while cartilage alteration (modified Mankin's score (mMs)) and targeted spinal pain neuropeptide were quantified upon sacrifice. Compared with the sedentary-placebo (presenting allodynia from D7), the exercise-placebo group showed an increase in sensitivity threshold (p < 0.04 on D7, D21, and D56). PA treatment was efficient on D56 (p = 0.001) and presented a synergic anti-allodynic effect with exercise from D21 to D56 (p < 0.0001). Histological assessment demonstrated a detrimental influence of exercise (mMs = 33.3%) compared with sedentary counterparts (mMs = 12.0%; p < 0.001), with more mature transformations. Spinal neuropeptide concentration was correlated with sensory sensitization and modulation sites (inflammation and endogenous inhibitory control) of the forced mobility effect. The surgical MI-RAT OA model coupled with calibrated slight exercise demonstrated face and predictive validity, an assurance of higher clinical translatability.

Abstract 185: Visual Recovery after Pituitary Apoplexy: Role of Intervention Timing and Treatment Modality

Introduction Pituitary apoplexy is a serious, emergent, and potentially dangerous condition of the pituitary in which the gland is affected by hemorrhage and/or ischemia typically in the setting of an underlying adenoma. In the present study, we investigate whether early versus delayed surgical intervention (in terms of three categories: before versus after 48 hours, 72 hours, and 7 days, respectively) resulted in any differences in vision outcomes for patients experiencing pituitary apoplexy. Furthermore, we compare the efficacy of surgical decompression versus conservative management as treatments for pituitary apoplexy. Methods Accordingly, we queried three databases in adherence to PRISMA guidelines. Quantitative meta‐analysis was performed according to the Mantel‐Haenszel method using Review Manager v5.4. P‐values &lt; 0.05 were considered statistically significant. Results Twenty‐nine studies remained eligible for review of vision outcomes follow pituitary apoplexy, including 16 studies for timing of intervention and 15 studies comparing intervention modality. For the cohort of patients featured in the studies describing intervention timing, there was a total of 322 patients, of which 269 (83.6%) identified as male. Most patients (56%‐100%) presented with a visual deficit, and all patients underwent surgery – most commonly via an EEA approach – before or shortly after the 2‐day, 3‐day, or 7‐day cutoff points. 220 patients were included in the sub‐analysis comparing patients who underwent surgery short of 7 days to those receiving treatment after 7 days following the onset of pituitary apoplexy. Furthermore, 81 patients underwent surgical decompression of the sella prior to 48 hours, whereas 32 patients underwent surgical decompression after 48 hours. Rutkowski’s 2017 study was the only to assess outcomes prior to versus after intervention at the 72 hours timepoint. Almost all patients in this study exhibited improved vision post‐decompression, including 19/19 (100%) in the post‐72‐hour cohort. On meta‐analysis using the Mantel‐Haenszel method, there was a significant difference in favor of patients who underwent surgical decompression before 7 days as compared to after seven days (OR 5.88 [1.77, 19.60], I^2=0%, P=0.004). Furthermore, there was a total of 288 patients across the 15 studies comparing surgical versus conservative management of pituitary apoplexy – which proved equivocal in select patient upon meta‐analysis. Conclusion In the present study, timing of surgical intervention for pituitary apoplexy was only found to be predictive of visual function recovery at the 7‐day timepoint, as has been reported by previous studies. Ultimately, this suggests that timeliness of intervention may have a significant impact on one of the most significant and potentially devastating implications of PA: vision loss. However, it does not appear that the optimal outcome is obtained by taking the PA patient to surgery “as soon as possible”. It is possible that timing of surgical intervention is a minimally critical or non‐critical factor in management of PA, and this should be taken into consideration when developing guidelines for treatment of PA. With regard to intervention modality, both surgery and conservative management appear to afford similar outcomes in most cases.

Gender differences in the antihypertensive effect after radiofrequency denervation of the renal arteries in resistant arterial hypertension

Objective: evaluation of gender differences in the antihypertensive effect after denervation of the renal arteries based on the results of office and daily measurements of blood pressure in resistant arterial hypertension in a 3-year follow-up.Materials and methods: the study involved 80 patients with resistant arterial hypertension (RAH); Patients were divided by stratification randomization into the interventional treatment group, who continue to receive antihypertensive therapy (AHT) (RDN group) and the MT group (patients on drug therapy only. The study groups were comparable in terms of gender, age, duration of hypertension, the number of antihypertensive drugs taken, the presence of For 12 months, a comparative study was conducted between groups, further analysis of the effectiveness of surgical treatment of RDN PA was carried out at control points of 24 and 36 months, in which laboratory and instrumental examinations were performed.Results: according to the office measurement of blood pressure within the RDN and MT study groups, both in men and women, it turned out to be similar. After 12 months of observation between the comparison groups, the women of the group showed a statistically significant negative dynamics of the office SBP (p&lt;0.01) and office DBP (p&lt;0.05) parameters. According to the analysis of average daily, average daily, average night SBP indicators, after 12 months in the RDN group, a decrease was observed in all main parameters (p&lt;0.001). It is worth paying attention to the fact that in the MT group, among the average daily, average daily, average night ABPM indicators after 12 months, no statistically significant changes were detected. When conducting a gender analysis of ABPM indicators using the Friedman test and correcting the p value for multiple comparisons at visits of 24 and 36 months, a statistically significant decrease in the main average daily indicators in women was demonstrated, while in men such changes were not established.Conclusions: the data obtained indicate a more pronounced antihypertensive effect after radiofrequency denervation of the renal arteries in resistant arterial hypertension in women.

The Role of Parkin in Rat Pancreatic Beta Cells Fate.

Parkin is a member of the mitochondrial quality control system that plays a major role in mitophagy. Although the loss of function mutations in the Parkin gene has been associated with the Familial Parkinson's phenotype, research in recent years points out that Parkin's function is not limited to neurodegenerative diseases. Parkin's function impressing key cellular quality control mechanisms, including the ubiquitin-proteasome and autophagy-lysosome systems, makes it an important player in the maintenance of cellular homeostasis. In this study, we investigated whether Parkin affects cell viability and ER stress responses under lipotoxic conditions in INS-1E cells. Our results may suggest that silencing Parkin may affect autophagy in addition to apoptosis. We also showed that Parkin may have a protective effect against lipo-toxic effects in INS-1E cells. Consistent with previous studies, we observed that stress responses were different for high and low palmitic acid doses. The Parkin being inhibited under high-dose PA treatment and active under low-dose PA treatment indicate that regulation of stress responses is controlled by environmental conditions. Our preliminary findings may suggest that in low lipotoxic conditions, Parkin affects the ER stress response by modulating Chop activity and Ca2+ release from the ER to the cytoplasm.

2-phenylacetamide Separated from the seed of Lepidium apetalum Willd. inhibited renal fibrosis via MAPK pathway mediated RAAS and oxidative stress in SHR Rats

BackgroundRenal fibrosis with Renin–angiotensin–aldosterone system (RAAS) activation and oxidative stress are one of the major complications in hypertension. 2-phenylacetamide (PA), a major active component of Lepidium apetalum Willd. (L.A), has numerous pharmacological effects. Its analogues have the effect of anti-renal fibrosis and alleviating renal injury. This study aims to explore the underlying mechanism of PA for regulating the renal fibrosis in SHR based on the MAPK pathway mediated RAAS and oxidative stress.MethodsThe SHR rats were used as the hypertension model, and the WKY rats were used as the control group. The blood pressure (BP), urine volume were detected every week. After PA treatment for 4 weeks, the levels of RAAS, inflammation and cytokines were measured by Enzyme-Linked Immunosorbnent Assay (ELISA). Hematoxylin–Eosin staining (HE), Masson and Immunohistochemistry (IHC) were used to observe the renal pathology, collagen deposition and fibrosis. Western blot was used to examine the MAPK pathway in renal. Finally, the SB203580 (p38 MAPK inhibitor) antagonism assay in the high NaCl-induced NRK52e cells was used, together with In-Cell Western (ICW), Flow Cytometry (FCM), High Content Screening (HCS) and ELISA to confirm the potential pharmacological mechanism.ResultsPA reduced the BP, RAAS, inflammation and cytokines, promoted the urine, and relieved renal pathological injury and collagen deposition, repaired renal fibrosis, decreased the expression of NADPH Oxidase 4 (NOX4), transforming growth factor-β (TGF-β), SMAD3 and MAPK signaling pathway in SHR rats. Meanwhile,,the role of PA could be blocked by p38 antagonist SB203580 effectively in the high NaCl-induced NRK52e cells. Moreover, molecular docking indicated that PA occupied the ligand binding sites of p38 MAPK.ConclusionPA inhibited renal fibrosis via MAPK signalling pathway mediated RAAS and oxidative stress in SHR Rats.

Dual effect of picolinic acid in the photosynthetic performance of rice plants infected by Bipolaris oryzae

AbstractBrown spot, caused by Bipolaris oryzae, is one of the most important diseases of rice. The non‐host toxin α‐picolinic acid (PA) has great potential to be used to enhance plant resistance against pathogen infection. The present study investigated the effect of spraying PA [0 (control), 1, 3, and 5 mg/mL] on the photosynthetic performance of rice plants (cultivar Metica‐1) infected or not with B. oryzae. Moreover, whether the PA treatment, especially at the highest concentration, could affect brown spot development was also evaluated. The chlorophyll a fluorescence parameters such as variable‐to‐maximum chlorophyll a fluorescence ratio (Fv/Fm), photochemical yield [Y(II)], yield for dissipation by down‐regulation [Y(NPQ)], the yield for non‐regulated dissipation [Y(NO)], and electron transport rate (ETR) as well as the concentration of photosynthetic pigments were determined. Based on the in vitro assay, PA inhibited mycelial growth of B. oryzae in a dose‐dependent manner and conidial germ tube length only decreased at 5 mg PA/mL. Conidia germination was not affected by the PA treatment. Necrotic lesions caused by PA were observed on leaves of non‐inoculated plants at 3 and 5 mg PA/mL. Symptoms of the brown spot were reduced on plants sprayed with 1 and 3 mg PA/mL compared to the control treatment. Brown spot lesions and those originating from PA toxicity overlapped for inoculated plants sprayed with 3 and 5 mg PA/mL. The photochemical performance of non‐inoculated plants was hampered by treatments with 3 and 5 mg PA/mL. Greater concentration of photosynthetic pigments and less impairment on the photosynthetic performance of inoculated plants sprayed with 1 mg PA/mL were noticed based on the values of Fv/Fm, Y(II), Y(NPQ), Y(NO), and ETR compared to inoculated plants non‐sprayed with PA. In conclusion, spraying rice plants with a low concentration of PA could decrease brown spot severity while preserving the photosynthetic capacity of the infected plants. The cellular damage generated by spraying the plants with the highest PA concentration did not favour the infection process of B. oryzae.

LGG-03. EXPLORING THE SIGNALING NETWORK INVOLVED IN MAPK PATHWAY INHIBITION BY THE MEK INHIBITOR TRAMETINIB IN BRAF-FUSION-DRIVEN PEDIATRIC PILOCYTIC ASTROCYTOMA

Abstract INTRODUCTION Pilocytic astrocytomas (PA) are the most common pediatric brain tumors. They are characterized by MAPK pathway alterations, leading to its constitutive activation and modulating the balance between proliferation and oncogene-induced senescence (OIS) sustained by senescence-associated secretory phenotype (SASP). Little is known about the molecular implications of MAPK pathway inhibition in the proliferating and senescent tumor compartments. METHODS DKFZ-BT66 cells derived from a primary KIAA:BRAF-fusion positive PA cell line and BT40 cells derived from pleomorphic xanthoastrocytoma with a BRAFV600E mutation and CDKN2A/B deletion, were used as model systems. RNA-sequencing and phospho-/proteomic datasets were generated in both the proliferative and senescent cells, and treated with the MEKi trametinib for different time-spans. A multi-omics factor analysis tool (MEFISTO) was used to identify key OIS/proliferation effectors. RESULTS Differential gene expression analysis revealed that MEK inhibition leads to the inhibition of the OIS/SASP gene program in senescent DKFZ-BT66. In addition, the protein level of several SASP factors was decreased. This translated in reduced sensitivity towards senolytics drugs, indicating inhibition of senescence features upon MEKi. MEFISTO analysis allowed to identify key transcription factors, genes and proteins involved in MAPK-induced OIS in the senescent PA cells, that were mapped using a prior knowledge network approach. Finally, single sample geneset enrichment analysis showed that most MAPK-related signatures were downregulated upon MEKi treatment, while pathways related to upstream MAPK activators (including several RTK pathways) were predicted to be upregulated, in both proliferating and senescent cells. CONCLUSION This data suggests that MAPKi reverses OIS in senescent PA cells, while inducing the activation of MAPK upstream regulators, identifying putative co-targets for the treatment of PA. Further validation of the targetability of these pathways is pending. Furthermore, the identification of the MAPK-related OIS/SASP genes provide insight about the regulation of OIS/SASP by the MAPK pathway.

GLP‑1 receptor agonist protects palmitate-induced insulin resistance in skeletal muscle cells by up-regulating sestrin2 to promote autophagy

In this study, we aimed to determine whether liraglutide could effectively reduce insulin resistance (IR) by regulating Sestrin2 (SESN2) expression in L6 rat skeletal muscle cells by examining its interactions with SESN2, autophagy, and IR. L6 cells were incubated with liraglutide (10–1000 nM) in the presence of palmitate (PA; 0.6 mM), and cell viability was detected using the cell counting kit-8 (CCK-8) assay. IR-related and autophagy-related proteins were detected using western blotting, and IR and autophagy-related genes were analyzed using quantitative real-time polymerase chain reaction. Silencing SESN2 was used to inhibit the activities of SESN2. A reduction in insulin-stimulated glucose uptake was observed in PA-treated L6 cells, confirming IR. Meanwhile, PA decreased the levels of GLUT4 and phosphorylation of Akt and affected SESN2 expression. Further investigation revealed that autophagic activity decreased following PA treatment, but that liraglutide reversed this PA-induced reduction in autophagic activity. Additionally, silencing SESN2 inhibited the ability of liraglutide to up-regulate the expression of IR-related proteins and activate autophagy signals. In summary, the data showed that liraglutide improved PA-induced IR in L6 myotubes by increasing autophagy mediated by SESN2.

Effects of water additive probiotic, Pediococcus acidilactici on growth performance, feed utilization, hematology, gene expression and disease resistance against Aspergillus flavus of Nile tilapia (Oreochromis niloticus)

Probiotics are live beneficial Micro-organism introduced into the gastrointestinal tract through food or water, probiotics confer several benefits and play important roles in improving growth performances, disease resistance, immunity, health status, gut microbiome, and water quality. The present study examined the effects of water additive probiotic, Pediococcus acidilactici, on growth performance, feed utilization, hematology, gene expression, and disease resistance in tilapia Oreochromis niloticus fingerlings challenged with fungus (Aspergillus flavus). Triplicate groups of Nile tilapia fingerlings (20.0 ± 0.4 g) were fed a commercial diet in four water treatments control (0 g/m3), 0.1PA (0.1 g//m3), 0.2PA (0.2 g/m3) and 0.3PA (0.3 g/m3) of P. acidilactici respectively. The water quality parameters particularly total ammonia nitrogen (TAN) and ammonia (NH3) decreased significantly (P ≤ 0.05) in all three water treatments 0.3PA treatment was the lowest) when compared to the control treatment. The highest significant values (P ≤ 0.05) of growth performance, feed utilization and hematology recorded of either 0.2PA or 0.3PA group, in addition, the expression of innate immune genes were significantly upregulated (P ≤ 0.05) of tilapia fingerling at 0.3PA group compared to the control. The results also showed that juveniles reared in three water treatments with additive probiotics had a significantly lower (P ≤ 0.05) mortality when challenged with Aspergillus flavus. Hence, the water additive probiotic used in this study is a promising solution as an alternative product to improve the water quality, growth, and health of O. niloticus.

Adipsin alleviates cardiac microvascular injury in diabetic cardiomyopathy through Csk-dependent signaling mechanism

BackgroundMicrovascular complications are associated with an overtly increased risk of adverse outcomes in patients with diabetes including coronary microvascular injury which manifested as disruption of adherens junctions between cardiac microvascular endothelial cells (CMECs). However, particular mechanism leading to diabetic coronary microvascular hyperpermeability remains elusive.MethodsExperimental diabetes was induced in mice with adipose tissue-specific Adipsin overexpression (AdipsinLSL/LSL-Cre) and their respective control (AdipsinLSL/LSL). In addition, cultured CMECs were subjected to high glucose/palmitic acid (HG + PA) treatment to simulate diabetes for a mechanistic approach.ResultsThe results showed that Adipsin overexpression significantly reduced cardiac microvascular permeability, preserved coronary microvascular integrity, and increased coronary microvascular density. Adipsin overexpression also attenuated cardiac dysfunction in diabetic mice. E/A ratio, an indicator of cardiac diastolic function, was improved by Adipsin. Adipsin overexpression retarded left ventricular adverse remodeling, enhanced LVEF, and improved cardiac systolic function. Adipsin-enriched exosomes were taken up by CMECs, inhibited CMECs apoptosis, and increased CMECs proliferation under HG + PA treatment. Adipsin-enriched exosomes also accelerated wound healing, rescued cell migration defects, and promoted tube formation in response to HG + PA challenge. Furthermore, Adipsin-enriched exosomes maintained adherens junctions at endothelial cell borders and reversed endothelial hyperpermeability disrupted by HG + PA insult. Mechanistically, Adipsin blocked HG + PA-induced Src phosphorylation (Tyr416), VE-cadherin phosphorylation (Tyr685 and Tyr731), and VE-cadherin internalization, thus maintaining CMECs adherens junctions integrity. LC-MS/MS analysis and co-immunoprecipitation analysis (Co-IP) unveiled Csk as a direct downstream regulator of Adipsin. Csk knockdown increased Src phosphorylation (Tyr416) and VE-cadherin phosphorylation (Tyr685 and Tyr731), while abolishing Adipsin-induced inhibition of VE-cadherin internalization. Furthermore, Csk knockdown counteracted Adipsin-induced protective effects on endothelial hyperpermeability in vitro and endothelial barrier integrity of coronary microvessels in vivo.ConclusionsTogether, these findings favor the vital role of Adipsin in the regulation of CMECs adherens junctions integrity, revealing its promises as a treatment target against diabetic coronary microvascular dysfunction.Graphical Graphical abstract depicting the mechanisms of action behind Adipsin-induced regulation of diabetic coronary microvascular dysfunction.

Effects of Dietary L-Proline and L-Alanine on Growth Performance, and Flesh Quality of Common Carp (Cyprinus carpio) Juveniles

Abstract The purpose of the present study was to investigate the effects of amino acids L-proline and L-alanine on growth performance, amino acid, and fatty acid levels in the fillet of juvenile common carp. Four hundred and fifty juvenile common carp were randomly distributed in 30 tanks and fed with three levels of proline (5 [P5], 10 [P10], and 15 [P15] g/kg), three levels of alanine [A] (5 [A5], 10 [A10], and 15 [A15] g/kg), three levels of proline-alanine combination [PA] (2.5 + 2.5 [2.5PA], 5 + 5 [5PA], 7.5 + 7.5 [7.5PA] g/kg feed) and basal diet (control). The highest body weight gain (25.85 ± 0.1 g) and survival rate (91.11 ± 3.84) were observed in 7.5 PA treatment. The highest protein content (64.58 ± 0.49) was noticed in 2.5 PA; however, compared to combined treatments (5 PA and 2.5 PA), it did not show any significant difference (P&lt;0.05). The highest total amount of essential amino acids (48.30 ± 48.3) was observed in 7.5 PA treatment. The highest amount of DHA (5.65 ± 0.08), total EPA, and DHA (8.91 ± 0.13) were in 7.5 PA treatment. Finally, it can be concluded that two amino acids L-proline and L-alanine at the combined level of 5 PA can improve the growth performance, survival, and fillet composition in the juvenile common carp.