Serum Activity Research Articles

Total antioxidant capacity status in non-obese adolescent females with PCOS: a cross-section study

Abstract Background The endocrine disease polycystic ovarian syndrome (PCOS) has a number of complications. Teenagers all over the globe are becoming increasingly concerned about the syndrome. It has been suggested that oxidative stress plays a significant role in the pathogenesis of PCOS. Aim The purpose of the present work was to estimate total antioxidant capacity (TAC) in non-obese adolescents and decide if the TAC could be a potential marker for the prediction and diagnosis of PCOS. Methods Serum samples were collected to assess the levels of follicle stimulant hormone (FSH), luteinizing hormone (LH), serum lipids, and serum total antioxidant capacity (TAC) in 50 non-obese patients with PCOS and 50 controls; correlation analysis was made between serum lipid, hormonal parameters, and TAC. Results Adolescents with PCOS had significantly lower levels of TAC compared with controls and showed significant higher values of LH, FSH, and LH/FSH ratio. There were significant negative correlations between TAC levels with LH, FSH, and LH/FSH. Linear regression analysis revealed statistically significant correlations between TAC and LH and FSH levels in non-obese adolescents with PCOS. Conclusion Serum TAC level was diminished in non-obese PCOS patients and was related to increase of LH/FSH ratio. So, quantification of TAC activity in serum could be of value in assessment of cases at higher risk for development of PCOS, suggesting its possible role in regulating sex hormones and could have potential roles in the etiopathogenesis of PCOS in non-obese adolescent females.

Lactobacillus paracasei 259 alleviates hyperuricemia in rats by decreasing uric acid and modulating the gut microbiota

Hyperuricemia (HUA) is a metabolic disease arising from abnormal purine metabolism. It contributes to an increased risk of kidney damage. The present study aimed to investigate the uric acid (UA)-lowering effects of Lactobacillus paracasei 259 isolated from yak yogurt and explore its underlying mechanisms. Our results revealed that L. paracasei 259 decreased the UA levels in rats and inhibited the serum activities of xanthine oxidase. In addition, L. paracasei 259 reduced the levels of pro-inflammatory cytokines (tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6) in the kidney and altered the expressions of UA transporters (ABC transporter 2 (ABCG2), PDZ domain containing 1 (PDZK1), urate transporter 1 (URAT1), and sodium-phosphate cotransporter type 4 (NPT4)) to near normal levels. Moreover, it increased the abundance of beneficial bacteria in the gut and recovered the gut microbiota composition, promoting the production of short-chain fatty acids (SCFAs). These findings suggested that L. paracasei 259 can potentially be used to decrease UA levels, repair kidney damage, regulate gut microbiota, and alleviate HUA.

Efficacy of Feed-Based Genome-Free Bacterial Vaccine Against Aeromonas hydrophila Infection in Red Tilapia (Oreochromis sp.)

Aeromonas hydrophila causes motile Aeromonas septicemia (MAS), a disease with a high mortality rate in tilapia culture. Feed-based vaccines with the incorporation of inactivated whole-cell bacteria into the feed offer promising tools to control MAS. Currently, the incorporation of genome-free bacteria as bacterial vaccine through the implementation of SimCells® technology into the feed has become a particular interest. Background/Objectives: This study investigates the efficacy of a feed-based vaccine incorporating genome-free A. hydrophila (FBV-GFAH) against MAS infection in red tilapia. Methods: The vaccine was prepared and delivered at 5% fish body weight for three consecutive days in weeks 0 (prime vaccination) and 2 (first booster vaccination), orally. Throughout a five-week experimental period, the immune-related genes (IL-1β, MHC-II, CD4, IgT, and IgM) expression in the hindgut and head kidney of the fish was determined using RT-qPCR assay. Lysozyme (serum) and overall IgM (serum, gut lavage, and skin mucus) productions were also detected. Results: Fish vaccinated with FBV-GFAH showed a significant (p ≤ 0.05) improvement in relative percent survival compared with unvaccinated fish following bacterial challenge. FBV-GFAH induced the expression of immune-related genes in the hindgut and head kidney, especially after booster vaccination. Furthermore, serum lysozyme activity and overall IgM production in serum, skin mucus, and gut lavage were also significantly (p ≤ 0.05) improved in the FBV-GFAH vaccinated fish than the unvaccinated fish. Conclusions: This study showed that FBV-GFAH is a promising feed-based vaccine technology to control MAS in cultured tilapia.

Biochemical Studies of Averrhoa carambola Ethanol Leaf and Fruit Extracts on Alloxan-induced Diabetic Rats

The use of herbal medicines has always been an option to treat diseases such as diabetes cancer and other ailments. This study aimed to evaluate the effect of Averrhoa carambola leaf and its fruit ethanol extracts on biochemical parameters and histopathology features. Forty five albino rats were assigned into 9 groups of 5 rats each. Except for the normal control group, others were induced to be diabetic by a single intraperitoneal injection of alloxan. Group 1 (normal control) and Group 2 (diabetic untreated) received water. Group 3 received 2.5mg/kg glibenclamide, Groups 4-9 received graded doses (250mg/kg, 500mg/kg and 750mg/kg) of leaf and fruit extracts of Averrhoa carambola. Treatment lasted 14days. At the end of the treatment, blood samples and organ (pancreas) were collected from the animals for biochemical and histopathological evaluation. Treatment of alloxan-induced diabetic rats significantly (p<0.05) reduced the glucose concentration across all treated groups. Following induction of diabetic mellitus, the serum activities of liver enzymes (Alanine amino transferase, Aspartate aminotransferase and Alkaline phosphatase), low-density lipoprotein cholesterol (LDL-C) and triglyceride were significantly (p<0.05) elevated in diabetic untreated group with corresponding reduction in serum protein, albumin and high-density lipoprotein cholesterol (HDL-C) compared to the normal control and treated groups. However, the fruit extracts exhibited a high increase in urea and creatinine levels especially at dose of 750mg/kg body weight when compared with the leaf though not significantly higher than the diabetic untreated group. From histopathology studies, the groups treated with the leaf extract of Averrhoa carambola showed greater structural improvement and an increase in pancreatic islet cells. Therefore this study showed that although ethanol extracts of Averrhoa carambola leaf and fruit have antidiabetic properties, the leaf extract possesses more antidiabetic effect.

A Novel Cell- and Virus-Free SARS-CoV-2 Neutralizing Antibody ELISA Based on Site-Specific Labeling Technology.

The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) led to the global spread of coronavirus disease 2019 (COVID-19), creating an urgent need for updated methods to evaluate immune responses to vaccines and therapeutic strategies. In this study, we introduce a novel cell-free, virus-free SARS-CoV-2 neutralizing antibody ELISA (NAb-ELISA), which is based on competitive inhibition of the receptor binding domain (RBD) of spike protein binding to the angiotensin-converting enzyme 2 (ACE2) receptor. In this method, site-specific biotinylated hACE2-Fc-Avi recombinant protein is immobilized onto a 96-well plate for capture, and the RBD-Fc-vHRP recombinant proteins serve as detection probes. Evaluation of sera from wild type (WT) or Delta RBD-immunized mice using the NAb-ELISA and pseudovirus neutralization tests (pVNTs) demonstrated strong correlations between assays (R2 = 0.91 and 0.90 for the WT and Delta groups, respectively). Additionally, the NAb-ELISA successfully detected cross-neutralizing activity in sera, though with slightly lower correlation to pVNT (R2 = 0.70-0.83). By employing NAb-ELISA instead of an indirect ELISA for hybridoma screening, five monoclonal antibodies (mAbs) with neutralizing activities against WT, Delta, and BA.2 pseudoviruses were obtained. This assay offers a straightforward, rapid, and safe approach to characterizing vaccine-induced antibody responses and mAb neutralization activity. Notably, the NAb-ELISA platform can be quickly adapted to assess neutralizing antibody responses against emerging mutant strains, addressing the rapid mutation of the virus.

Serum Glutamate dehydrogenase activity enables sensitive and specific diagnosis of hepatocellular injury in humans.

Serum activities of alanine- and aspartate- aminotransferases (ALT and AST) are considered the "gold standard" biomarkers of hepatocyte injury in clinical practice and drug development. However, due to expression of ALT and AST in myocytes, the diagnosis of hepatocellular injury in patients with underlying muscle diseases, including drug-induced muscle injury, is severely limited. Thus, we proposed glutamate dehydrogenase (GLDH) as a liver-specific alternative to serum ALT and AST. In fact, our exploratory studies showed that GLDH has comparable performance to ALT for detecting hepatocyte injury without interference from concomitant muscle injury. Here, we report the results of studies confirming the reference intervals in a healthy human population and sensitivity and specificity of GLDH for detection of hepatocyte injury in human subjects. In human subjects, we could not perform liver biopsies due to ethical reasons, we also confirmed the relationship of GLDH and histopathologic lesions using 32 model toxicants in rats. Furthermore, we have shown that injury to tissues that are known to express appreciable levels of GLDH does not affect serum GLDH measurements, indicating excellent liver specificity of serum GLDH. Finally, we observed faster elimination of GLDH than ALT in humans, indicating that decreasing GLDH values could be considered an early sign of recovery. This study provides comprehensive evidence of excellent sensitivity and liver specificity of GLDH for diagnosis of hepatocellular injury, including evaluation of reference intervals that is essential for interpretation of serum GLDH in human subjects.

Effects of Replacing Fish Meal with Rubber Seed Cake on Growth, Digestive, Antioxidant and Protein Metabolism of Juvenile Asian Red-Tailed Catfish (Hemibagrus wyckioides).

An 8-week feeding trial was conducted to assess the impact of replacing fish meal (FM) with rubber seed cake (RSC) on growth performance, digestive enzyme activity of the gut, antioxidant capacity and protein metabolism-related parameters of plasma and liver in juvenile Asian red-tailed catfish (Hemibagrus wyckioides) with an average body weight of 3.21 g. Four isonitrogenous (41.08 ± 0.17) and isolipidic (10.07 ± 0.11) diets were formulated, incorporating RSC at a level of 0%, 15%, 30%, and 45% (w/w) to progressively replace FM. Dietary inclusion of 15-30% RSC did not significantly affect growth performance (p > 0.05). However, the replacement of FM with RSC resulted in linear reductions in intestinal trypsin and amylase activities, with a significant reduction in intestinal amylase activity observed at the 45% RSC inclusion level. Dietary inclusion of 30-45% RSC decreased plasma glutamate dehydrogenase and aspartate aminotransferase activities, but increased plasma adenosine monophosphate deaminase and hepatic γ-glutamyltransferase activities. This may indicate that the high substitution of FM by RSC may affect the homeostasis of amino acid metabolism. Dietary inclusion of 15% RSC did not affect serum and hepatic antioxidant enzyme activities; however, dietary inclusion of 45% RSC significantly increased plasma malondialdehyde level, but decreased catalase activity. Furthermore, dietary inclusion of 30-45% RSC significantly increased hepatic lactate dehydrogenase activity. These results demonstrated that replacing a portion of FM with RSC is feasible, and dietary inclusion of 15% RSC did not have significant adverse effects on growth performance, digestive function and antioxidant capacity of H. wyckioides.

Mulberry galacto-oligosaccharides ameliorates non-alcoholic fatty liver disease in mice induced by a high-fat diet

A type of oligosaccharide named mulberry galacto-oligosaccharides (MGO), was isolated from mulberry with a galactose-only, weighs 987 Da and displays diverse biological properties. The objective of this work was to investigate the protective effects of MGO in non-alcoholic fatty liver disease (NAFLD) mice. The results showed that MGO can reduce the body weight and liver weight of NAFLD mice, and reduce the levels of NEFA, TG, TC and LDL-C in serum and liver as well as the activities of AST and ALT. Increase HDL-C content, SOD, and GSH activity in serum and improve pathological damage. MGO regulates the AMPK lipid metabolism pathway, upregulates the expression of ATGL, downregulates the expression of SREBP-1, inhibits lipogenesis, and promotes lipolysis. MGO alleviates liver function damage in NAFLD mice, improves blood lipids, liver metabolism, and increases the body’s antioxidant capacity, suggesting MGO potential as a dietary supplement and functional food to combat HFD-induced NAFLD.

Docosahexaenoic acid alleviated liver lipid deposition and health damage induced by cholesterol accumulation in hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂)

Cholesterol accumulation can be a critical pathological marker and, while, the cholesterol-lowering potential of docosahexaenoic acid (DHA) has been documented extensively in mammals, its effects in fish are unclear. The present study investigated the adverse effects of cholesterol accumulation on liver health, and evaluated DHA as a potential cholesterol-lowering agent to mitigate liver damage induced by high cholesterol (HC) intake in hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂). Grouper were fed either a control diet or an HC (1.6 %) diet supplemented with varying levels of DHA (0, 0.5, 1, 2 %) for eight weeks. The HC diet significantly increased condition factor (CF), mesenteric fat index (MFI), and lipid, cholesterol and triglyceride accumulation in liver. Supplementing the HC diet with 0.5 % DHA significantly reduced CF, MFI and liver lipid contents. Moreover, HC intake increased aspartate transaminase (AST) and alanine transaminase (ALT) activities in serum, and malondialdehyde (MDA) content and the expression of gene related to inflammatory responses and apoptosis in liver. In addition, HC intake significantly reduced mitochondrial biogenesis and total antioxidant capacity in liver. However, supplementing the HC diet with 0.5 % DHA significantly decreased liver cholesterol accumulation by inhibiting cholesterol synthesis and promoting cholesterol efflux, and significantly reduced liver triglyceride content by inhibiting the expression of genes associated with lipogenesis and lipid droplet formation. Furthermore, supplementation of the HC diet with 0.5 % DHA significantly reduced the activities of ALT and AST in serum, and MDA content and the expression of genes associated with inflammation, apoptosis and antioxidation in liver, and enhanced liver mitochondrial biogenesis. Overall, the results indicated that DHA supplementation mitigated liver cholesterol accumulation by inhibiting cholesterol synthesis and promoting cholesterol efflux, and enhanced mitochondrial function, boosted antioxidant capacity, and reduced inflammation in liver of grouper fed the HC diet, ultimately alleviating liver damage caused by cholesterol accumulation. The present study elucidated the detrimental effects of cholesterol accumulation on liver health in aquatic animals and proposed DHA as a potential intervention for mitigating health issues associated with cholesterol overload.

PRELIMINARY ASSESSMENT OF THE ACUTE TOXICITY EFFECTS OF FRIED CAT FISH EXTRACT ON BIOCHEMICAL, HEMATOLOGICAL AND HISTOPATHOLOGICAL PARAMETERS OF WISTAR RATS

Some food processing techniques have been linked with formation of toxic chemicals known as Polycyclic Aromatic Hydrocarbons (PAHs), which are found in some processed foods and have been widely attributed to the prevalence of cancer of liver and stomach in Nigeria. However, the toxicological effects of the consumption of fried fish have not been adequately documented. Therefore, this study investigated the toxicological effects of PAHs in fried cat fish on Wistar rats. Fifty five rats were used in the study and they were divided into control and ten treatment groups with daily administration of fried fish extract at dosage of 200, 400, 600, 800 and 1000 mg/kg body weight to the rats for 14 days. The results showed that the activities of serum, alanine aminotransferase (ALT), alkaline phospherase (ALP), asparte aminotransferase (AST) as well as urea and creatinine concentration were affected across the treated groups of animals which were indications for hepatorenal damage. White blood cell (WBC) and mean corpuscular volume (MCV) showed significant changes in response to the administered dosage compared to the control. The liver showed no significant histopathological changes between the test rats and control. The architecture of sections of kidney treated groups was similar to that of the control in which the renal corpuscles maintained their normal size of urinary space and normal tubular structures with no necrosis observed. The result revealed that there can be potential toxicity if there is repeated indulgence to fried cat fish at the temperatures between 160 – 230 oC.

Alginate-chitosan hydrogel formulations for VEGFA expressed baculovirus delivery promoting angiogenesis for wound healing and revascularization

Abstract Background Baculoviruses, engineered to express growth factors, offer a promising avenue for short-term gene therapy, such as wound dressings, due to their safety and specificity. Encapsulation in natural polymers like alginate and chitosan addresses limitations like serum inactivation and fragility, while promoting sustained delivery and shielding from immune inactivation. Wound healing involves complex processes that can be enhanced by maintaining an antimicrobial, moist environment, which promotes cell migration and angiogenesis. Vascular endothelial cell growth factor A (VEGFA) is known to be one of the most potent pro-angiogenic factors and plays a key role in wound healing. Purpose This investigation seeks to enhance wound healing and angiogenesis, support the revascularization process, and provide anti-inflammatory and anti-microbial effects. Additionally, it aims to assess the preclinical efficacy and safety of the approach. Methods Ionic cross-linking was used for virus encapsulation under aseptic conditions. The capsules were assessed for their surface morphology, particle size, zeta potential, and in vitro release profile. Additionally, their therapeutic potential was studied using cell lines. The efficacy of hydrogel delivery of VEGFA-expressing baculoviruses in promoting cell migration and angiogenesis for wound healing applications was investigated on Human Umbilical Vein Endothelial Cells (HUVECs). Hydrogel morphology, swelling, and encapsulation efficiency were evaluated, along with endothelial tube formation assays and hemocompatibility studies on HUVECs. Results Encapsulation of baculovirus expressing the VEGFA gene in alginate and chitosan–alginate hydrogels achieve a high encapsulation efficiency of 99.9%. This allows for prolonged virus delivery and an increased therapeutic window. The encapsulated baculovirus maintains activity and is released over eight days, with a transduction efficiency of over 40%. This promotes tube formation, cell proliferation, and cell migration for angiogenesis, particularly beneficial for chronic wound-healing applications. The hydrogels also prevented E. coli and C. albicans growth directly below and inhibited C. albicans growth surrounding the alginate–chitosan hydrogels. The hydrogels demonstrated no cytotoxicity and good blood compatibility. Conclusion This proof of concept underscores the potential of encapsulated baculovirus delivery systems for various gene therapy applications including cardiovascular tissue regeneration and revascularization.

Short-term reduction in feed intake by dairy cows in the postpartum period leads to subclinical ketosis development

Subclinical ketosis is widespread in highly productive dairy cows after calving and often remains undiagnosed, leading to reduced productivity. Physiologically controlled feeding in the first weeks after calving and during the intensive lactation period can reduce the incidence of ketosis. The study aimed to determine how a short-term reduction of feed after calving affects the formation of ketone bodies in blood, urine and milk of dairy cows. The group of ten Ukrainian black-spotted dairy breed cows after calving aged from 4 to 6 years was involved in the experiment lasted for 72 h. In 24 h the amount of compound feed, haylage, and silage received by cows was gradually reduced until complete exclusion in the diet. Blood, urine, and milk samples were collected three times a day. The content of ketone bodies, β-hydroxybutyrate and glucose was estimated with the corresponding sets of indicator strips. The content of total bilirubin, cholesterol, albumin and enzymes activity in the blood serum were determined on biochemical analyzer. It was found that in 24 h after the beginning of feed reduction the level of blood glucose decreased, persistent hypoglycemia within 48 and 72 h was developed, the concentration of β-hydroxybutyrate in blood and milk and of ketone bodies in urine was elevated. The increase in total bilirubin concentration and liver enzymes activity in the blood serum with a simultaneous decrease in albumin level and delayed sodium propionate conversion into glucose were observed. Thus, short-term reduction in feed intake by dairy cows after calving causes ketosis development and violation of liver functions. Keywords: dairy сows, feeding level, glucose, ketosis, liver damage, β-hydroxybutyrate

The effect of a probiotic drug on an enterosorbent on the change in the functional activity of humoral factors of nonspecific immunity of the Russian Lena sturgeon (Acipenser gueldenstadtii × Acipenser baeri)

In modern aquaculture, probiotic drugs are used to increase the immunologically determined natural resistance of the fish body, regulate their immune homeostasis, prevent stress, prevent diseases and form normal intestinal microflora of fish at all stages of development. The use of these drugs in compound feeds can solve a number of problems related to the immunological and physiological state of the fish body, affecting the antimicrobial activity expressed by the secretion of antimicrobial peptides, suppression of bacterial invasion, and prevention of adhesion of pathogenic microorganisms to epithelial cells. In addition, the drugs will help to improve the barrier function, expressed by an increase in mucus production, strengthening the integrity of the epithelial barrier. This article presents information about the experimental study of the effect of a probiotic drug on the enterosorbent “Ecoflor” on the dynamics of bactericidal activity of blood serum, the concentration of nonspecific immune complexes in blood serum and immunocompetent organs, the number of immunodeficient individuals, as well as on the level of C-reactive protein of the Russian Lena sturgeon (Acipenser gueldenstadtii x Acipenser baeri). It was found that the use of a probiotic drug in experimental dosages of 5–9 g/kg of compound feed has an effect on the indicators of nonspecific humoral immunity. Thus, the Russian- Lena sturgeon, grown on compound feeds with probiotic on enterosorbent, had an increased level of BABS and the absence of IMD of individuals. In addition, the probiotic contributed to a decrease in the concentration of nonspecific immune complexes in the blood serum and immunocompetent organs of the Russian-Lena sturgeon, and the results of the analysis for SRB allowed us to conclude that the drug used does not cause inflammatory reactions in the body of sturgeon fish.

A Practical Study on the Relationship between Serum Iron Elevation and Liver Enzymes Plus Other Factors in Patients with Beta-Thalassemia

Thalassemia is a genetic condition characterized by faulty hemoglobin synthesis and low hemoglobin concentration. Overload of iron is an unavoidable condition experienced by severe thalassemia patients as a result of excessive blood transfusions. We looked at impaired liver function in thalassemia individuals with Iraq as a result of these issues. Our findings demonstrated serum activity in hepatocytes, as well as a large rise in GOT and AIP in patients with elevated iron levels and a substantial rise in TSB. All of these elements may work together or independently to create chronic liver disease by causing damage in cells via oxidative mechanisms.

Exploring the Relationship between Canine Paraoxonase-1 (PON-1) Serum Activity and Liver Disease Classified by Clinico-Pathological Evaluation.

Paraoxonase-1 (PON-1), a liver-synthesized enzyme, acts as a negative acute-phase reactant during systemic inflammation in dogs. Given the hepatic synthesis of this enzyme, the presence of liver diseases may influence PON-1, thus affecting its reliability as a biomarker for inflammatory/oxidative systemic diseases. The aim of this study is to investigate PON-1 activity variations among dogs suspected of liver injury or failure, evaluating the influence of hepatic diseases on PON-1 activity. A total of one-hundred-sixty dogs were retrospectively enrolled and categorized into three groups based on clinical presentation and laboratory results: control (C = 20), suspected liver injury (INJ = 114), and suspected liver failure (FAIL = 26). The INJ group was further divided into subgroups based on the severity of the alanine aminotransferase (ALT) increase. Both the INJ and FAIL groups were further divided based on serum macroscopic appearance. The PON-1 activity was quantified using a paraoxon-based method, which is already validated in dogs. No significant difference in PON-1 activity was observed between the C and INJ groups, despite a significant increase in the subgroups with moderate and severe elevations of ALT. The dogs with icteric serum exhibited decreased PON-1 activity, while lipemic serum was associated with an increased PON-1 activity. A significant reduction in PON-1 activity was noted in the FAIL group, compared to both C and INJ groups (p < 0.0001), regardless of serum appearance. Given the retrospective nature of this study, additional evaluations (e.g., histopathology, imaging) were not performed. The results obtained here suggest the importance of interpreting PON-1 activity cautiously in dogs with suspected liver disease.

Palygorskite improves growth performance and prevents liver damage in avian pathogenic Escherichia coli-challenged broiler chickens at an early age.

Avian pathogenic Escherichia coli (APEC) is a major bacterial infection that causes economic losses in the global poultry industry. Palygorskite (PAL) has been shown to enhance growth performance while improving antioxidative and anti-inflammatory properties of broilers. This study evaluated the protective effects of PAL on growth performance and liver function in broilers subjected to APEC challenge. A total of 320 one-day-old male Arbor Acres chicks were divided into four groups with eight replicates of ten birds each, based on a 2×2 factorial arrangement (basal diet or 5g/kg PAL-supplemented diet) and inoculation (bacterial culture medium or APEC). PAL increased body weight gain (BWG) prior to APEC challenge (P < 0.05). However, APEC caused losses in BWG, feed intake (FI), and feed efficiency, along with increased relative hepatic weight, hepatic pathology scores, and hepatic-cell apoptosis rate (P < 0.05). Compared to normal birds, APEC increased interleukin (IL)-1β, interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), malondialdehyde (MDA), and nitric oxide (NO) levels, as well as lysozyme (LZM) and myeloperoxidase (MPO) activities, while decreasing total antioxidant capacity (T-AOC) and IL-10 levels, and total superoxide dismutase (T-SOD) and catalase (CAT) activities in both serum and liver, APEC also raised alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, but reduced total protein (TP), albumin (ALB), immunoglobulin (Ig) A, IgG, and IgM levels in serum (P < 0.05). Moreover, APEC increased hepatic mRNA level of IL-1β, IFN-γ, TNF-α, nuclear factor kappa B, and inducible nitric oxide synthase (iNOS), while inhibited mRNA level of IL-10 (P < 0.05). In contrast, PAL increased BWG and FI, alleviated hepatic-cell apoptosis rate during the challenge period (P < 0.05). An incorporation of PAL reduced triglyceride and NO contents, ALT and AST activities, while increasing TP, ALB, IL-10, IgG, and IgM levels in serum, enhancing serum T-SOD and CAT activities, elevating hepatic T-AOC and CAT activities, inhibiting hepatic MDA accumulation, and reducing IL-1β levels and LZM activity in both liver and serum (P < 0.05). An interactive effect was found for hepatic TNF-α and iNOS mRNA expression, in which PAL inhibited their mRNA expression in APEC-challenged birds (P < 0.05). Overall, PAL addition partially mitigated the negative impact of APEC challenge on growth performance and liver function of broiler chicks at an early age.

Qiwei Jinggan Ling regulates oxidative stress and lipid metabolism in alcoholic liver disease by activating AMPK

BackgroundAlcoholic liver disease (ALD) is a severe public health concern worldwide and there is still a lack of effective treatments. Qiwei Jinggan Ling (QJL) has protective effects against various liver injuries, but its pharmacological action on ALD has received little attention. PurposeTo investigate the effect and mechanism of QJL on ALD in vivo and in vitro. MethodsIn vivo, an ALD mouse model was established by alcohol combined with a high-fat diet (HFD) and treated with QJL. Biochemical indicators, HE staining, and Oil Red O staining were employed to assess hepatic oxidative stress, steatosis, and alcohol metabolism. RNA sequencing analysis was performed, and the results were verified by qRT-PCR and Western blot to elucidate the hepatoprotective mechanism of QJL. In vitro, HepG2 cells were co-stimulated with NaOA (sodium oleate) and EtOH (ethanol), followed by intervention with Compound C (CC, AMPK inhibitor) and QJL-containing serum. Oil Red O, BODIPY (boron-dipyrromethene), and ROS (reactive oxygen species) staining were applied to validate the efficacy and mechanism of QJL-containing serum. The expression of AMP-activated protein kinase (AMPK) pathway-related factors was analyzed through qRT-PCR and Western blot for additional corroboration. Moreover, the key pharmacodynamic components of QJL were identified by UPLC-MS/MS and molecular docking. ResultsIn vivo, QJL ameliorated liver structural disorders, steatosis, oxidative stress, and impaired alcohol metabolism, as indicated by biochemical indicators and histopathological assays. RNA sequencing analysis revealed that QJL reversed the expression of genes related to alcohol metabolism, fatty acid metabolism, and cholesterol metabolism. The results of qRT-PCR and Western blot were in line with those of RNA sequencing. Furthermore, it was discovered that QJL significantly upregulated the expression of p-AMPK and downregulated the expression of sterol regulatory element binding transcription factor 1 (SREBP-1c). In vitro, biochemical indicators and staining assays demonstrated that QJL-containing serum inhibited lipid accumulation and oxidative stress. The qRT-PCR and Western blot analysis revealed that QJL-containing serum markedly enhanced the expression of p-AMPK and carnitine palmitoyltransferase 1a (Cpt1a), while suppressing the expression of SREBP-1c, fatty acid synthase (Fasn), and acetyl-coenzyme A carboxylase 1 (ACC-1). However, CC inhibited the above pharmacological activities of QJL-containing serum. Additionally, (2S)-Liquiritigenin, Glycyrrhetinate, Isovitexin, Taxifolin, and Yohimbine were proved to be the key active components of QJL. ConclusionQJL had the potential to be a therapeutic drug for ALD by activating the AMPK pathway, thereby regulating lipid metabolism and inhibiting oxidative stress.

Serotype 15C Streptococcus pneumoniae resistant to classical complement deposition and agglutination by polyclonal rabbit anti-capsular 15B sera

BackgroundS. pneumoniae (SP) serotypes 15B and 15C are frequent causative agents of invasive pneumococcal disease (IPD), otitis media, and nasopharyngeal colonization in the post PCV13 era. The principal difference between serotypes 15B and 15C is the presence of an O-acetyl group on the pentasaccharide repeating unit of 15B polysaccharide. It remains unclear if antibodies against SP15B polysaccharide demonstrate functional cross reaction with SP15C strains. We compared functional activity of polyclonal rabbit anti-capsular 15B sera against SP15B and SP15C isolates. MethodsUsing flow cytometry we measured complement factors C3c and C4d deposition on SP15B and 15C in the presence of polyclonal rabbit anti capsular 15B sera. We measured the binding of C3c, common to all complement pathways, and C4d, specific to classical pathway, on SP serotypes 15B and 15C when co-incubated with polyclonal rabbit anti capsular 15B sera and antibody depleted complement. Both the proportion of bacteria with complement deposition and the fluorescence intensity were measured. We also measured agglutination as the increase in forward and side scatter. ResultsPolyclonal rabbit anti-capsular 15B sera activated classical pathway resulting in deposition of C4d and C3c at high intensity on all SP15B cells but only achieved limited deposition and intensity of C4 with SP15C. Similarly, polyclonal rabbit anti-capsular 15B sera induced agglutination of SP15B strains in a dose dependent manner and limited agglutination of SP15C. ConclusionsAnti-capsular 15B sera induce limited C4d and C3c deposition, and minimal agglutination with SP15C strains, reflecting lower classical pathway activation in contrast to high C4d and C3c deposition and agglutination of SP15B. These observations support limited functional cross reactivity of anti-15B to SP15C strains and are consistent with the reduction in opsonophagocytic killing of SP15C reported following immunization with vaccines containing 15B polysaccharide.

A-083 Performance evaluation of Sentinel Diagnostics’ ACE assay on the Abbott Alinity c system and ARCHITECT c16000 System

Abstract Background The analytical performances of the Angiotensin Converting Enzyme (ACE) assay for quantitative determination of ACE activity in human serum and plasma have been evaluated on the Abbott Alinity c system and ARCHITECT c16000 System. ACE (dipeptidyl carboxypeptidase) is a glycoprotein peptidyldipeptide hydrolase that cleaves histidylleucine dipeptide from angiotensin I, a relatively inactive decapeptide. The latter is converted to the potent vasoconstrictor, angiotensin II. ACE also inactivates bradykinin. Elevated levels of ACE activity occur in serum of patients with active sarcoidosis, and occasionally in premature infants with respiratory distress syndrome, in adults with tuberculosis, Gaucher’s disease, leprosy, and in many other pathologic conditions involving lung and liver diseases. Methods ACE hydrolyses urylacryloylphenylalanine-glycylglycine (FAPGG) to furylacryloylphenylalanine (FAP) and glycylglycine. Hydrolysis of FAPGG results in a decrease in absorbance at 340 nm. The ACE activity in the sample is determined by comparing the sample reaction rate to that obtained with the Sentinel ACE calibrator. Performances evaluation have been done following the latest CLSI guidelines protocols available. Results Summary of the analytical performances for the tested assay is reported in table below. Conclusions Analytical performances of Sentinel Diagnostics’ ACE assay demonstrate that quantitative determination of ACE activity in human serum and plasma are suitable for the routine measurement of the analyte on Abbott Alinity c system and ARCHITECT c16000 System.

Determined Transglutaminase 1 Activity in Blood Serum for Brain Cancer Patients and Healthy People

Cancer is defined as a group of abnormal cells that grow and spread unusually, and sometimes their proliferation cannot be controlled. These tumours are usually called cancer. Cancer may affect damaged parts of the body, including the brain. This study focused on the activity of transglutaminase 1 enzyme in the blood serum of brain cancer patients and then compared it with the control group. The study also included the effect of age, gender, body mass index, and glucose on the transglutaminase 1 enzyme in the blood serum of patients with brain cancer. The study sample consisted of 45 samples of brain cancer patients, (45) samples as a control group. The results of this study showed a significant decrease in the activity of transglutaminase 1 in brain cancer patients (902.66 ± 14.27 ng/L) compared to the control group (1594.66 ± 32.95 ng/L). Gender and age also impact the enzyme; in the control group only, males had lower enzyme activity than females, and the enzyme's activity decreased with age in both groups (brain cancer patients and controls). The findings also demonstrate that there was a discernible difference in the serum glucose concentration of brain cancer patients when compared to the control group, although it was still within normal allowable limits, and that there was a decrease in the enzyme's activity with an increase in body mass index values in both the patient and control groups. The investigation of transglutaminase 1 is a useful marker for identifying brain cancer and can be utilized in early diagnosis.