Spleen Weight Research Articles

Allergic Disposition of IVF-Conceived Mice

With the increased utilization of assisted reproductive technology (ART), concerns about the potential health risks for ART-conceived babies have also been raised. Increased prevalences of allergic and metabolic diseases have been reported among ART offspring. This study aimed to evaluate the impact of IVF on the tendency to develop allergic responses following ovalbumin (OVA) sensitization in IVF-conceived mice. Mice were divided into four groups (non-OVA naturally conceived, OVA naturally conceived, non-OVA IVF-conceived, and OVA IVF-conceived). In the OVA groups, the mice were subjected to intraperitoneal and intranasal immunization with OVA. Two days after the final immunization, blood samples were taken, and the serum levels of IgE and IL-4 were detected by ELISA. The mice were sacrificed by cervical dislocation, their spleens and lungs were removed, and their weights were measured and recorded. Sensitization with OVA resulted in significantly increased concentrations of IL-4 and total IgE, as well as increased lung and spleen weights, among offspring from both natural and IVF conception. The concentrations of IgE and IL-4 and the lung and spleen weights in IVF-conceived mice were significantly higher compared to those in naturally conceived mice before and after sensitization with OVA. It is concluded that compared to naturally conceived mice, IVF-conceived mice exhibit a greater tendency to develop allergic responses against OVA.

Soy Protein and Safflower-Seed Oil Attenuate Inflammation and Immune Dysfunction in Rats with Hyperuricemia

A plant-based diet is considered a promising approach for managing hyperuricemia (HUA). This study examined the effects of soy protein and plant-based oils on HUA-induced inflammation and immune dysfunction. Male Wistar rats, induced with HUA using oxonic acid and uric acid (UA), were fed casein or soy protein with palm or safflower oil (2 × 2 factorial design) for 8 weeks. HUA rats had lower serum albumin and T cell percentages in peripheral blood leukocytes (PBLs) and splenocytes, along with increased leukocyte counts and spleen weights, compared to healthy rats (p < 0.05). Soy protein improved HUA-induced reductions in albumin, while safflower-seed oil ameliorated reductions in albumin, plasma interleukin (IL)-4, and T-suppressor splenocytes, and mitigated elevated serum UA, plasma IL-6, and B leukocytes (two-way ANOVA, p < 0.05). In PBL, soy protein alleviated HUA-induced decreases in TNF-α, casein and palm oil increased IL-6, and casein further reduced IFN-γ production. Under Con A stimulation, casein and safflower-seed oil alleviated decreases in IL-6 and IL-10, respectively, while under LPS stimulation, casein further increased TNF-α production. In splenocytes, soy protein and safflower-seed oil reduced HUA-induced increases in TNF-α and increased IL-10, and safflower-seed oil increased IL-6 production. Under Con A stimulation, soy protein and safflower-seed oil reduced TNF-α and increased IL-10 production in splenocytes. The findings suggest that soy protein and safflower-seed oil may counteract HUA-related inflammation, alleviate monocyte activation, and enhance Th2 immune response in HUA. A plant-based diet rich in soy protein and safflower-seed oil may help manage HUA and associated inflammation and immune dysfunction.

Inhibition of alternative complement system and prolyl hydroxylase ameliorates anaemia of inflammation.

Chronic diseases associated with inflammation cause early destruction of RBCs. Complement system, part of innate immunity, is involved in such RBC destruction. Persistent inflammation causes kidney injury, leading to reduced erythropoietin release and functional iron deficiency, causing anaemia. We have investigated effect of iptacopan, a factor B inhibitor, and desidustat, a prolyl hydroxylase inhibitor in anaemia induced by peptidoglycan polysaccharide (PGPS) treatment in rats. Inflammation, haemolysis and its diagnostic markers (LDH, total bilirubin, and RBC lifespan) were evaluated after three days of PGPS challenge. Haemoglobin, RBC, iron homeostasis, and RBC destruction were evaluated fourteen days after PGPS challenge. Desidustat (15mg/kg) and iptacopan (20mg/kg) were given along with PGPS and continued for two weeks. Iptacopan and its combination with desidustat prevented LDH, total bilirubin, complement protein-C3a and haemolysis. Combination treatment caused an early normalization of haemoglobin and RBC. Combination also reduced WBC, alkaline phosphatase, aspartate aminotransferase, and rat paw volume. Serum iron was increased by desidustat and its combination treatment. Spleen weight, tissue iron, and serum hepcidin were reduced by combination treatment. Effect of desidustat alone was prominent on iron (serum and tissue) and hepcidin. Thus, combination of iptacopan and desidustat can be a potentially useful therapeutic option for treatment of anaemia of inflammation.

ORGAN-SPARING AND ORGAN- REMOVING SURGICAL PROCEDURES FOR PATHOLOGICAL CONDITIONS OF THE SPLEEN IN CHILDREN. QUESTIONS AND ANSWERS.

Introduction. Nowadays splenic surgery is a rather urgent problem in pediatric surgery. There are diseases where only splenectomy allows a child to live a full life. These are: sickle cell anemia (SCA), idiopathic thrombocytopenic purpura, hereditary spherocytosis (Minkowski- Chauff ard disease), thalassemia, and autoimmune hemolytic anemia. The spleen is involved in 25-30 % of cases of blunt abdominal trauma in children. The loss of more than 45 % of the circulating blood volume leads to an unstable hemodynamic state in the child. The question of surgical treatment and preservation of the spleen in hemoperitoneum due to its injury is of vital importance. Aim: To study the possibilities of surgical treatment of diseases and injuries of the spleen in children. Materials and methods. An analysis of surgical interventions for diseases and injuries of the spleen in children aged 2 to 17 years from 2019 to 2024 was performed. The study was conducted in the surgical departments of the «Clinical Center of Pediatric Medicine» of the «Okhmatdyt Hospital» of the «Western Ukrainian Specialized Center» (Lviv) and the «Chernivtsi City Children’s Clinical Hospital» (Chernivtsi). 27 children (17 girls, 10 boys) had spleen diseases and 43 children (23 girls, 20 boys) had spleen injuries. Splenectomy was performed for the following splenic diseases: sickle cell anemia (9 children), idiopathic thrombocytopenic purpura (4 children), hereditary spherocytosis (Minkowski- Chauff ard disease) (6 children), thalassemia (1 child), and autoimmune hemolytic anemia (1 child). Six children had splenic cysts: 3 children had posttraumatic cysts and 3 children had parasitic cysts. Twenty children underwent surgery for splenic injuries. The following surgical procedures were performed: laparoscopic abdominal cavity repair with spleen preservation followed by conservative therapy (13 children); laparoscopic splenectomy (2 children); laparotomy, splenic suture (2 children); and open splenectomy (3 children).Results. Conservative management of splenic injuries of varying degrees according to the American Association for the Surgery of Trauma (AAST) was performed in 23 (53.49 %) children. Resorption of hemoperitoneum in cases of conservative treatment of splenic injuries occurred 11-14 days after the start of treatment. Safe, full physical activity was resumed after 6 weeks of conservative treatment of splenic injury. Surgical procedures were performed in 20 (46.51 %) children using various techniques. 17 patients underwent laparoscopy. 2 children underwent laparoscopic splenectomy for grade IV AAST injury and hemodynamic instability. In 15 cases, treatment was started with conservative therapy, but during monitoring an increase in hemoperitoneum was observed, indicating the need for laparoscopy. Laparoscopy was performed in 13 children. A minimum of 100 ml and a maximum of 400 ml of blood was collected laparoscopically in the children. The abdominal cavity was examined and hemostasis was confi rmed. In cases where the spleen was «wrapped» with the omentum, the omentum was not separated from the spleen. The operation was completed by placing a drainage tube in the perisplenic space. In 2 children, grade IV AAST injuries were converted to an open procedure in which the spleen was sutured to stop bleeding and a drain was left near the splenic bed. Grade V AAST splenic injury in 3 children was an indication for left subcostal laparotomy and splenectomy. For splenic disease, laparoscopic splenectomy was performed in 12 (44.44 %) children, with high effi cacy in Minkowski- Chauff ard disease combined with gallstones (3 children) where simultaneous cholecystectomy was possible. Laparoscopic removal of splenic cysts was also justifi ed in 3 (11.11 %) children. However, removal of parasitic cysts required open splenectomy in 3 patients. Open splenectomy was performed in 9 children with splenic disease. In 5 children the inappropriateness of laparoscopy was due to severe perisplenitis, large spleen weight and young age of the children.Conclusions1. The morpho- functional structure of the spleen promotes spontaneous hemostasis, which is the basis for conservative treatment of spleen injuries in 53.49 % of children.2. The AAST classifi cation of splenic injuries from grade I to V, hemodynamic indicators and ultrasound monitoring of blood in the abdominal cavity can be used as a basis for choosing the method of treatment of closed blunt splenic injuries in children.3. Ineff ectiveness of conservative treatment, hemodynamic instability and increase of blood in the abdominal cavity in a child with splenic injury within 4 hours is an indication for laparoscopy.4. Splenic suturing and laparoscopic monitoring should be the preferred methods for treating closed splenic injuries in children to preserve the organ.5. The choice of surgical intervention for splenic diseases in children should be individualized and depend on the disease, the child’s age, the size of the spleen, and the presence of complications of the disease.

Effect of terracotta drinker and/or water addition of ASPRO‐C plus on behavior, growth, and physiological response of broiler chickens exposed to high temperature

AbstractThis study aimed to evaluate the effects of a terracotta drinker and/or water supplementation with ASPRO‐C Plus on the zootechnical performance of broiler chickens reared in a hot environment. A total of 160 Cobb 500 broiler chicks of 15 days old (240.2 ± 39.82 g) were divided into four treatment groups in a 2 x 2 factorial arrangement of drinker type (plastic or terracotta) and water with or without ASPRO‐C Plus (1 g/L) supplementation, each consisting of 4 replicate pens. Respiratory rate, water intake, feed intake, and live body weight were recorded weekly. At 49 days old, 12 birds per group were randomly selected, fasted for 12 h, weighed, and slaughtered for carcass evaluation and blood collection. The respiration rate of broilers decreased significantly (p < 0.01) with the terracotta drinker as compared to the plastic drinker. The water intake, the feed intake and the body weight gain increased significantly (p < 0.01) with the terracotta drinker as compared to the plastic drinker. The water addition of ASPRO‐C Plus significantly increased (p < 0.01) the relative weight of abdominal fat and spleen in broilers. The alpha‐amylase activity was significantly decreased (p < 0.01) with the water addition of ASPRO‐C Plus. The serum content of total cholesterol was significantly increased (p < 0.01) with the terracotta drinker. It can be concluded that ASPRO‐C Plus can slightly improve liveability, but using the terracotta drinker can be more efficient in reducing the behavioral response to heat stress and can improve the growth performance.

The Protective Effect of Esculentoside A on MPL/lpr Mice by Upregulating the Expression of CD19+IL-35+Breg Cells and Interleukin-35

Introduction: Esculentoside A (EsA) is one of the main components of the traditional Chinese medicine Phytolacca esculenta. The possible mechanism of action of EsA in the treatment of lupus nephritis (LN) was explored by observing the effects of EsA on CD19+ IL-35+regulatory B (IL-35+Breg) cells. Methods: Twenty-four MRL/lpr mice were randomly divided into control, EsA, and EsA+IL-12p35 antibody groups. Mice were administered the respective treatments intraperitoneally once a day for 4 weeks. The urine protein/creatinine ratio (UPCR) and blood creatinine (Cr) and IL-35, IL-10, and IL-17 expression levels were measured. Body and spleen weight were measured to calculate the splenic index (SI). Flow cytometry was performed to determine the proportion of CD19+ IL-35+ Breg cells in the spleen. Hematoxylin-eosin and PASM-Masson staining of renal tissues were performed, and the “Austin” acute index (AI) system for LN was determined. Results: The most severe conditions were seen in mice in the control group, with the highest UPCR, Cr, and IL-17 levels and SI and AI scores; the most severe renal histopathology, and the lowest proportion of CD19+ IL-35+ Breg cells and IL-35 and IL-10 levels. This was followed by the EsA+IL-12p35 antibody group. The EsA group had the lowest UPCR, Cr, and IL-17 levels and SI and AI scores; the mildest renal lesions; and the highest proportion CD19+ IL-35+ Breg cells and IL-35 and IL-10 levels. Conclusion: EsA delayed the progression of LN by promoting the proliferation of CD19+ IL-35+ Breg cells, upregulating the expression of IL-35, and decreasing the secretion of IL-17.

Effects of Centella asiatica-isolated pectic polysaccharide on dextran sulfate sodium-induced colitis

Dietary supplementation of polysaccharides demonstrates strong therapeutic actions on inflammatory bowel disease (IBD). Centella asiatica (CA) has traditionally been used in Ayurveda and Chinese medicine. However, the effects of CA-isolated pectic polysaccharides on IBD remain unknown. This study determined the effect of a CA-isolated pectic polysaccharide on IBD. The crude polysaccharide (CA-CP), isolated from a hot-water extract of CA, is a typical pectic polysaccharide composed mainly of galacturonic acid (40.6 %), galactose (27.6 %), arabinose (13.5 %), and rhamnose (8.5 %). CA-CP improved clinical symptoms in a DSS-induced colitis murine model, including weight change (9.9–12.0 %), disease activity index (31.6–51.9 %), colon length (13.2–21.5 %), and spleen weight (21.8–26.3 %). CA-CP effectively regulated the levels of inflammatory and junctional factors by mediating the MAPK and NF-κB pathways. CA-CP partially alleviated DSS-induced crypt destruction, submucosal edema, inflammatory infiltration, and mucin secretion. The content of total short-chain fatty acids increased substantially (cecum 29.8–53.7 %, feces 75.4–109.3 %) with oral CA-CP administration compared to the DSS group. Cecal microbial community analysis revealed that CA-CP administration regulated DSS-induced colitis by reducing the abundance of Escherichia (5.0–10.9 %) and Clostridium (0.3–0.4 %), while increasing the abundance of Bacteroidetes (6.8–8.5 %), Ligilactobacillus (2.7–6.0 %), and Bilophilia (0.3–0.6 %). The findings provide fundamental data for developing novel functional therapeutic agents for the prevention and treatment of colitis, using CA-isolated pectic polysaccharides. Furthermore, to the best of our knowledge, our study is the first to demonstrate the effects of pectic polysaccharides isolated from CA on IBD.

Caloric Vestibular Stimulation Alleviates Cold Water Swimming Stress Induced Changes in BDNF and Haematological Parameters

Introduction: Stress is inevitable in daily life. Stress in long term has been implicated in the development of physical and neuronal disorders. Various studies have described the levels and role of BDNF in the brain following stress. Studies representing the changes in serum BDNF and wet weight of organs in stress are limited and controversial. Aim: The aim of the present study is to find the effect of stress on serum BDNF concentration, wet weight of selected organs, haematological parameters and to evaluate the effectiveness of caloric vestibular stimulation on stress-induced changes in these parameters. Methods and material: Male Wistar rats were exposed to cold water swimming stress for 14 days for a duration of 30 minutes per day. Serum corticosterone and serum BDNF were measured using the ELISA method. Weight of liver, spleen, thymus and lymph nodes were measured and their relation to body weight was calculated. Total RBC count, total WBC count, platelet count and haemoglobin concentration were analysed. Results: A statistically significant increase in serum corticosterone and BDNF was observed following stress. Liver weight, thymus weight and lymph nodes weight did not show any significant changes but spleen weight has reduced in the normal recovery group following stress. Total RBC count, total WBC count, platelet count and haemoglobin levels were significantly higher in the stress group. Caloric vestibular stimulation was effective in managing stress-induced changes in serum corticosterone, BDNF and spleen weight. Conclusion: Caloric vestibular stimulation is effective in restoring the stress-induced changes.

In Silico and In Vivo Evaluation of Anti-Arthritic Effects of Bakuchiol from Psoralea corylifolia Seeds in Experimental Rat Model.

Rheumatoid arthritis is an autoimmune disease mainly affecting the joints categorized by inflammation, swelling of the synovium and decrease in the joint movement. Bakuchiol, a meroterpene class of natural product present in Psoralea corylifolia known to possess anti-inflammatory effects by a variety of mechanisms. However, its effects in rheumatoid arthritis still remain unclear. In the present investigation, we studied the anti-arthritic effects of bakuchiol via in silico and in vivo experiments. It also showed antioxidant effects measured using DPPH assay where it showed free radical scavenging activity with IC50 value 468.26 μg/ml. Molecular Docking studies carried out on COX-1 (PDB ID: 3 N8Z), COX-2 (PDB ID: 4PH9) and TNF-α (PDB ID: 7JRA), proteins involved in inflammation in arthritis. Bakuchiol showed the maximum binding affinity for TNF-α with binding affinity score is -7.29 kcal/moland less affinity was observed for COX-1 and 2. In vivo antiarthritic effects were studied in arthritic female wistar rats model prepared by intradermal injection of freund's complete adjuvant. Bakuchiol was administered orally at dose of 10,20 and 40 mg/kg for 21 days. Our treatment showed that bakuchiol at 20 and 40 mg/kg exhibited significant anti-inflammatory effects (p<0.001) showed by significant decrease in paw volume, paw diameter, spleen and thymus weight and increase in pain threshold and body weight in arthritic rat model. A significant decrease in hematological parameters such as total leukocyte count (TLC), platelet count, CRP and rheumatoid arthritis factor (RF) and increase in red blood cells count, ESR and hemoglobin further demonstrated that bakuchiol treatment suppresses the progression of adjuvant induced arthritis (AIA) in arthritic rat model. Histological analysis further revealed that bakuchiol ameliorates the pathological manifestations of AIA and reverse the abnormality induced by AIA in rats shown by protection against bone necrosis involved with low influx of inflammatory cells. Therefore, in silico and in vivo results revealed that bakuchiol has the potential to be developed as potent antiarthritic agent.

Wire injury induced moderate aortic valve stenosis in mice is accompanied by a chronic systemic inflammatory reaction

Abstract Background Patients with aortic valve stenosis (AS) have elevated serum levels of inflammation markers, and blood cytokine levels correlate with ventricular recovery after transcatheter aortic valve replacement. While presence of inflammatory processes in stenotic aortic valves is acknowledged, no systematic characterization of the systemic immune reaction upon AS has been performed, yet. Purpose Hypothesis of this study was that AS induces a systemic inflammatory reaction linked with local processes in the heart. Methods AS was induced by wire injury (WI) or sham surgery was performed in 3-4 months old male C57Bl/6J mice. AS severity, left ventricular (LV) function and hypertrophy indices were assessed with echocardiography (echo). After four weeks, explanted organs were weighted, and flow cytometry identified total leukocyte numbers in blood, heart, and peripheral immune organs (spleen, liver, lymph nodes) and uptake of Cy5-labelled perfluorocarbon nanoemulsions by whole blood leukocytes. Costimulatory molecules (flow cytometry, RT-qPCR) and cytokines (RT-qPCR, Multiplex Immunoassay) were analysed in heart, peripheral immune organs and blood. Results R-values were positive for spleen weight correlation with AS severity, cardiac function and mass suggesting cardiac hypertrophy to preserve heart function. Myocardial myeloid and T cell r-values were positive or negative, respectively, for correlation with spleen and liver weight, hinting at T cell recruitment from peripheral stores. Immune cell numbers in peripheral lymph nodes and spleen showed negative r-values for correlation with heart hypertrophy indices; number of liver myeloid cells correlated negatively with LV wall thickness (monocytes=-0.8571, p=0.0238) suggesting immune cell recruitment to hypertrophic hearts. Cytokine mRNA levels trended mainly towards increase in heart and regional lymph nodes and reduction in spleen and liver after WI. Correlation with echo was more homogeneous after WI, with r-values mainly positive, except of T cell activation markers, for aortic valves, but negative for myocardium, hinting at mechanisms preserving heart function. Correlating unchanged cytokine protein levels in myocardium and plasma with echo, r-values were mostly positive for myocardium, and were more positive than in sham for plasma. Echo and costimulatory molecule correlation showed a more homogeneous pattern in WI, with mostly positive r-values in myocardium and periphery, while most r-values were negative in sham. Together with reduced PFC-uptake by lymphatic cells, this hints at systemic immune cell activation in AS. Conclusion The results suggest that number and activation state of leukocytes in peripheral organs are directly linked to cardiac processes in AS. Considering the pathological value of inflammation, it is crucial to in future studies find out if modulation of the systemic inflammatory reaction relieves severity of AS and prevents development of heart failure.

Alterations in tissue content of iron and zinc in mice bearing hepatoma 22a and their correction by zinc sulphate supplementation

It is known that many tumors induce iron and zinc deficiency in the organism. We studied the content of these metals, as well as the specific activity of two antioxidant metal-dependent enzymes – catalase and superoxide dismutase of three distal organs (thymus, liver and spleen) in animals bearing transplantable hepatoma 22a. These alterations were compared to weight changes of organs. On day 21 of tumor growth, as compared to control group, nonheme iron content in all three organs was decreased, and zinc content – only in the thymus. The specific activities of catalase and superoxide dismutase were both increased in the thymus, while in the liver activity of superoxide dismutase decreased. At the same time point thymic involution and splenomegaly were developed. In order to normalize metal content mice bearing hepatoma 22a were supplemented with 22 mkg of zinc sulphate per ml of drinking water during 3 weeks. Zinc sulphate supplementation partly compensated zinc deficiency in the thymus, increased zinc content in the liver and restored iron content in three organs. It also normalized superoxide dismutase activity in the liver and had no influence on enzymes in other organs. Zinc supplementation did not influence the weight of spleen and liver, but prevented the development of thymic involution. Moreover, metal deficiency in the thymus was restored while the activity of antioxidant enzymes remained unchanged. Based on this we can conclude that thymus involution in hepatoma 22a mice was associated with iron and zinc deficiency in this organ and was not linked with antioxidant enzyme activity, while splenomegaly had no relation to both types of parameters in the spleen. Thus, zinc sulphate positively influences metabolism of two vital trace elements – zinc and iron in animals bearing hepatoma 22a, what contributes to maintaining of the central immune organ – the thymus, and along with this it improves antioxidant system of the liver.

Therapeutic oligonucleotide ASC1R shows excellent tolerability and remarkable efficacy in reducing SARS-CoV-2 mRNA levels in C57BL/6 mice

The coronavirus pandemic has resulted in over 775 million cases and 7 million deaths worldwide, driving efforts to develop therapeutic strategies to control the viral infection. Therapeutic oligonucleotides have shown promise in treating many pathological conditions, including those of viral origin. The present study assessed the in vivo efficacy and safety of ASC1R, a novel therapeutic oligonucleotide of unconventional design targeting the conserved viral RdRp sequence essential for replication. In functional studies, ASC1R was administered to transfected C57BL/6 mice at doses of 1 and 10 mg/kg. Safety assessments included acute toxicity evaluations at doses ranging from 30 to 100 mg/kg, and subacute toxicity evaluations of repeated doses of 1 and 10 mg/kg. Evaluations included general clinical observations, findings at necropsy, measurements of organ weight, and histopathological examinations of the liver, lungs, spleen, and kidneys. ASC1R effectively reduced RdRp levels >94 % within 24 hours following a single 1 mg/kg dose, with no observed organ toxicity. Acute and subacute toxicity assessments found that mice receiving high (≥30 mg/kg) or repeated (10 mg/kg for 7 days) doses of ASC1R showed an increase in relative spleen weight, without histopathological changes. The marked ability of a single low dose of ASC1R (1 mg/kg) to reduce viral RNA suggests its potential for clinical applications, balancing therapeutic efficacy with minimal side effects. Our findings indicate that ASC1R has promise as a viable treatment option for patients with COVID-19.

Cardiometabolic effects of sacubitril/valsartan in a rat model of heart failure with preserved ejection fraction

The promising results obtained in the PARADIGM-HF trial prompted the approval of sacubitril/valsartan (SAC/VAL) as a first-in-class treatment for heart failure with reduced ejection fraction (HFrEF) patients. The effect of SAC/VAL treatment was also studied in patients with heart failure with preserved ejection fraction (HFpEF) and, although improvements in New York Heart Association (NYHA) class, HF hospitalizations, and cardiovascular deaths were observed, these results were not so promising. However, the demand for HFpEF therapies led to the approval of SAC/VAL as an alternative treatment, although further studies are needed. We aimed to elucidate the effects of a 9-week SAC/VAL treatment in cardiac function and metabolism using a preclinical model of HFpEF, the Zucker Fatty and Spontaneously Hypertensive (ZSF1) rats. We found that SAC/VAL significantly improved diastolic function parameters and modulated respiratory quotient during exercise. Ex-vivo studies showed that SAC/VAL treatment significantly decreased heart, liver, spleen, and visceral fat weights; cardiac hypertrophy and percentage of fibrosis; lipid infiltration in liver and circulating levels of cholesterol and sodium. Moreover, SAC/VAL reduced glycerophospholipids, cholesterol, and cholesteryl esters while increasing triglyceride levels in cardiac tissue. In conclusion, SAC/VAL treatment improved diastolic and hepatic function, respiratory metabolism, reduced hypercholesterolemia and cardiac fibrosis and hypertrophy, and was able to modulate cardiac metabolic profile. Our findings might provide further insight into the therapeutic benefits of SAC/VAL treatment in obese patients with HFpEF.

Jianpi-Huatan-Huoxue-Anshen formula ameliorates gastrointestinal inflammation and microecological imbalance in chemotherapy-treated mice transplanted with H22 hepatocellular carcinoma.

Jianpi-Huatan-Huoxue-Anshen formula [Tzu-Chi cancer-antagonizing & life-protecting II decoction (TCCL)] is a Chinese medical formula that has been clinically shown to reduce the gastrointestinal side effects of chemotherapy in cancer patients and improve their quality of life. However, its effect and mechanism on the intestinal microecology after chemotherapy are not yet clear. To discover the potential mechanisms of TCCL on gastrointestinal inflammation and microecological imbalance in chemotherapy-treated mice transplanted with hepatocellular carcinoma (HCC). Ninety-six mice were inoculated subcutaneously with HCC cells. One week later, the mice received a large dose of 5-fluorouracil by intraperitoneal injection to establish a HCC chemotherapy model. Thirty-six mice were randomly selected before administration, and feces, ileal tissue, and ileal contents were collected from each mouse. The remaining mice were randomized into normal saline, continuous chemotherapy, Yangzheng Xiaoji capsules-treated, and three TCCL-treated groups. After treatment, feces, tumors, liver, spleen, thymus, stomach, jejunum, ileum, and colon tissues, and ileal contents were collected. Morphological changes, serum levels of IL-1β, IL-6, IL-8, IL-10, IL-22, TNF-α, and TGF-β, intestinal SIgA, and protein and mRNA expression of ZO-1, NF-κB, Occludin, MUC-2, Claudin-1, and IκB-α in colon tissues were documented. The effect of TCCL on the abundance and diversity of intestinal flora was analyzed using 16S rDNA sequencing. TCCL treatment improved thymus and spleen weight, thymus and spleen indexes, and body weight, decreased tumor volumes and tumor tissue cell density, and alleviated injury to gastric, ileal, and colonic mucosal tissues. Among proteins and genes associated with inflammation, IL-10, TGF-β, SIgA, ZO-1, MUC-2, and Occludin were upregulated, whereas NF-κB, IL-1β, IL-6, TNF-α, IL-22, IL-8, and IκB-α were downregulated. Additionally, TCCL increased the proportions of fecal Actinobacteria, AF12, Adlercreutzia, Clostridium, Coriobacteriaceae, and Paraprevotella in the intermediate stage of treatment, decreased the proportions of Mucipirillum, Odoribacter, RF32, YS2, and Rikenellaceae but increased the proportions of p_Deferribacteres and Lactobacillus at the end of treatment. Studies on ileal mucosal microbiota showed similar findings. Moreover, TCCL improved community richness, evenness, and the diversity of fecal and ileal mucosal flora. TCCL relieves pathological changes in tumor tissue and chemotherapy-induced gastrointestinal injury, potentially by reducing the release of pro-inflammatory factors to repair the gastrointestinal mucosa, enhancing intestinal barrier function, and maintaining gastrointestinal microecological balance. Hence, TCCL is a very effective adjuvant to chemotherapy.

Aflatoxins and fumonisins co-contamination effects on laying hens and use of mycotoxin detoxifiers as a mitigation strategy.

This study examined the effects of fumonisins (FBs) and aflatoxin B1 (AFB1), alone or in combination, on the productivity and health of laying hens, as well as the transfer of aflatoxins (AFs) to chicken food products. The efficacy and safety of mycotoxin detoxifiers (bentonite and fumonisin esterase) to mitigate these effects were also assessed. Laying hens (400) were divided into 20 groups and fed a control, moderate (54.6µg/kg feed) or high (546µg/kg feed) AFB1 or FBs (7.9mg/kg feed) added diets, either alone or in combination, with the mycotoxin detoxifiers added in selected diets. Productivity was evaluated by feed intake, egg weight, egg production, and feed conversion ratio whereas health was assessed by organ weights, blood biochemistry, and mortality. Aflatoxins residues in plasma, liver, muscle, and eggs were determined using UHPLC-MS/MS methods. A diet with AFB1 at a concentration of 546µg/kg feed decreased egg production and various AFB1-contaminated diets increased serum uric acid levels and weights of liver, spleen, heart, and gizzard. Interactions between AFB1 and FBs significantly impacted spleen, heart, and gizzard weights as well as AFB1 residues in eggs. Maximum AFB1 residues of 0.64µg/kg and aflatoxin M1 (below limits of quantification) were observed in liver, plasma, and eggs of layers fed diets with AFB1. The mycotoxin detoxifiers reduced effects of AFB1 and FBs on egg production, organ weights, blood biochemistry, and AFB1 residues in tissues. This study highlights the importance of mycotoxin detoxifiers as a mitigation strategy against mycotoxins in poultry production.

Ferroptosis-associated alterations in diabetes following ischemic stroke: Insights from RNA sequencing

ObjectiveFerroptosis is an iron-dependent form of programmed cell death associated with lipid peroxidation. Though diabetes worsens cerebral injury and clinical outcomes in stroke, it is poorly understood whether ferroptosis contributes to diabetes-exacerbated stroke. This study aimed to identify ferroptosis-associated differentially expressed genes in ischemic stroke under diabetic condition and then explore their roles using comprehensive bioinformatics analyses. MethodsType 1 diabetes (T1D) model was established in male mice at 8–10 weeks of age by one intraperitoneal injection of streptozotocin (110 mg/kg). Ischemic stroke was induced by a transient 45-minute middle cerebral artery occlusion and evaluated three days thereafter. Ischemic brain cortex was dissected 24 h after the reperfusion and subjected to bulk tissue RNA sequencing followed by bioinformatics analysis and verification of key findings via quantitative real-time PCR. ResultsEnlarged infarct size was seen in diabetic, as compared with non-diabetic mice, in conjunction with worsened neurological behaviors. Both body and spleen weights were reduced in diabetic as compared with non-diabetic mice. There was a trend for reduced survival rate in diabetic mice following the stroke. In RNA sequencing analysis, we identified 1299 differentially expressed genes in ischemic brain between diabetic and non-diabetic mice, with upregulation and downregulation for 732 and 567 genes, respectively. Among these genes, 27 genes were associated with ferroptosis. Further analysis reveals that solute carrier family 25 member 28(SLC25A28) and sterol carrier protein 2(SCP2) were the top genes associated with ferroptosis in diabetic mice following ischemic stroke. In several bioinformatics analyses, we found SLC25A28, one of the top ferroptosis-related genes, is involved in several metabolic and regulatory pathways as well as the regulatory complexity of microRNAs and circular RNAs, which demonstrates the potential role of SLC25A28 in diabetes-exacerbated stroke. Drug network analysis suggests SLC25A28 as a potential therapeutic target for ameliorating ischemic injury in diabetes. ConclusionsOur bulk RNA sequencing and bioinformatics analyses show that altered ferroptosis signaling pathway was associated with the exacerbation of experimental stroke injury under diabetic condition. Especially, additional investigation into the mechanisms of SLC25A28 and SCP2 in diabetes-exacerbated stroke will be explored in the future study.

Comparative Analysis of Drowning Index: Relevance in Drowning Deaths versus Non-Drowning Asphyxial Deaths

Drowning remains a significant cause of mortality worldwide, necessitating accurate diagnostic tools for forensic investigation. This paper conducts a comparative analysis of the Drowning Index (DI) to ascertain its relevance in distinguishing drowning deaths from non-drowning asphyxial deaths. Drowning deaths present a challenge for forensic pathologists, because the autopsy findings may occur in many non-drowning scenarios. Previous studies have attempted to identify patterns in organ weights that may be specific for drowning. The drowning index has been defined as the weight ratio of the lungs and pleural effusion fluid to the spleen. We compared the lung and pleural effusion weight, spleen weight, and DI from 124 autopsies of asphyxia deaths including both drowning deaths and non-drowning asphyxia deaths such as hanging, strangulation, suffocation and mechanical asphyxia at the department of Forensic Medicine and Toxicology at the Gandhi Medical College, Bhopal from January 2021 to August 2022. Findings highlight the potential utility of the DI as a valuable adjunctive tool in forensic investigations, providing insights into its comparative efficacy in differentiating drowning from other modes of asphyxial deaths. Such insights are critical for enhancing accuracy in forensic diagnoses and contributing to advancements in medicolegal death investigations.

In ovo betaine and heat acclimation affect hatching results, growth performance and immunity of the broilers under cyclicheatstress.

This study investigated the effects of in ovo betaine and thermal manipulation during incubation on growth performance, and some immune parameters of broilers under cyclic heat stress (CHS). Eggs were divided into 5 groups and incubated at 1) 37.8°C and 60% relative humidity (Control incubation and not-injected, CI); 2) eggs were incubated at CI and in ovo betaine injected into yolk sac on d 11 (E11) (CI + In ovo); 3) eggs were exposed to 38.8°C for 8h between 10 and 18d of incubation (heat acclimation, HA); 4) eggs were incubated at HA and in ovo betaine applied (HA + In ovo); 5) positive control: eggs were incubated at CI and injected with saline. Hatched chicks were raised under standard management conditions until 21 d, between 21 and 42 d half of the chickens in each incubation treatment were kept either at optimum (OPT) or at CHS. In ovo and HA did not affect hatching performance. In ovo increased thymus and spleen weights of chicks. Serum IgG was higher in HA + In ovo chicks. From d 7 to 21, in ovo chicks were heavier body weights, consumed more feed, and better feed conversion than those from CI. The body weights of HA chickens were similar in OPT and CHS on d 28 and 35. CHS reduced the body weight of CI chickens which was compatible with their feed consumption. Moreover, feed intake of HA + In ovo chickens exposed to CHS was higher than those not injected indicating that HA + In ovo enhanced thermoregulation of chickens under CHS.

Evaluation of inhibitory potential of the flavonoid-rich fraction of Myrica esculenta Buch.-Ham. ex D. Don against DSS-induced colonic inflammation in mice

ObjectiveMyrica esculenta (family Myricaceae), a valuable plant species in China and India, has been traditionally used by many tribes and local communities to manage gut disorders. Scientific validation of its anti-ulcerative colitis activity was aimed. MethodsThe ethyl acetate fraction of Myrica esculenta (MeEa) was prepared and evaluated for its potency against DSS-induced ulcerative colitis (UC) in mice at 200 and 400 mg/kg BW oral dose. The effective dose of MeEa was determined through its effect on DSS-induced UC and was further analyzed through its effects on disease activity index (DAI), colon length, colon weight/length ratio, spleen weight, serum and colon tissue cytokine level, cell count and hemoglobin content. Further, the effect was determined through histopathology and FITC-dextran-induced membrane permeability assay. ResultsBetween the two doses, MeEa at 400 mg/kg BW was found to be the most effective dose in terms of reduced DAI scores, protected colon length from shortening, decreased colon weight/length ratio, reduced spleen weight, decreased pro-inflammatory cytokine level and stabilized the anti-inflammatory cytokine level in serum and colon tissue. MeEa 400 reduced cell counts and increased hemoglobin content and platelet count. MeEa 400 also prevented the colon by protecting epithelial cells and crypts. MeEa 400 provided significant protection from intestinal leakage and reduced FITC dextran level in serum. DiscussionMeEa 400 possesses significant anti-inflammatory potential and acts via attenuation of DSS-induced UC and inhibition of DAI scores. It reduces pro-inflammatory cytokines and stabilizes anti-inflammatory cytokine levels, reduces cell count, and protects epithelial tissue and crypts in the colon, as well as intestinal membrane leakage that occurred due to FITC-dextran administration in mice.

8624 Maternal ZIKV Infection Causes Oxidative Damage in Placenta of Both Female and Male Mice

Abstract Disclosure: V.M. Nascimento: None. I.S. Andrade: None. C.B. Andrade: None. R.S. Alves: None. R.P. Sousa: None. A.C. Valim: None. L.S. Dias: None. K.N. Fontes: None. I.F. Miranda: None. S.A. Coelho: None. L.B. Arruda: None. R.S. Fortunato: None. T.M. Ortiga-Carvalho: None. The placenta is a maternal-fetal organ with nutritional, exchanging, hormonal and protecting functions that are vital for fetal growth and development during gestation. However, these functions can be disturbed by vertically transmitted pathogens such as Zika virus (ZIKV). In 2015, ZIKV was responsible for an increasing number of newborns with congenital abnormalities in Brazil, known as Congenital Zika Syndrome (CZS). Nonetheless, the extension of ZIKV infection in the placenta, leading to CZS, is still unclear. It is known that ZIKV causes oxidative stress in trophoblasts. However, no one has ever studied placental oxidative stress caused by ZIKV infection in vivo. Here, we investigated whether maternal ZIKV infection could cause sex-specific oxidative stress in murine placenta. At gestational day (GD)12.5, pregnant mice (C57BL/6; 8-12 weeks old) were infected with ZIKV-BRPE243 (5x107 PFU; ZIKV group, n=22) or Mock (supernatants of noninfected C6/36 cells; control group, n=28). At GD18.5, c-section was performed and maternal spleens, placentas, fetuses and fetal brains were collected. We measured placental carbonylated protein levels (n=7), by 2D OxyBlot, nitrotyrosine residues (n=5), by immunohistochemistry, reduced thiol content (n=9), total superoxide dismutase (SOD; n=9) and total glutathione peroxidase (GPx; n=9) activities, by spectrophotometry. Data were analyzed by unpaired Student’s t test and Two-Way ANOVA with Tukey’s post test and represented as mean±SEM. ZIKV infection increased maternal spleen weight (p&amp;lt;0.0001) and decreased both female (p=0.0429) and male (p=0.0171) placental weight. Although placental weights were affected by the infection, female and male fetal weight and fetal crown-rump length were not changed. Conversely, maternal ZIKV infection increased only female fetal brain weight (p=0.0028). Moreover, we found that ZIKV infection increased nitrotyrosine residues in the placental labyrinth region of both female (p&amp;lt;0.0001) and male (p=0.0007) fetuses. Meanwhile, nitrotyrosine residues were increased by the infection only in the placental junctional zone of female fetuses (p=0.0131). Although ZIKV increased this oxidative damage biomarker, placental carbonylated protein levels were not changed by the infection in both sexes. Also, reduced thiol content and total SOD and GPx activities were not affected. Together, our data show that maternal ZIKV infection caused oxidative stress in both placental regions of female and male fetuses, as we saw that ZIKV increased oxidative damage, although antioxidant defenses were not changed by the infection, at least at term (GD18.5). ZIKV infection may have disturbed placental growth, as we found that placental weight was decreased in both sexes. Moreover, we believe that the increase in the female fetal brain weight could be a suggestive feature of cerebral edema or hydrocephalus. Presentation: 6/2/2024