Blood Serum Of Mice Research Articles

Supplementation of syringic acid-rich Phrynium pubinerve leaves imparts protection against allergic inflammatory responses by downregulating iNOS, COX-2, and NF-κB expressions

BackgroundThe present study was designed to characterize the role of ethanolic leaf extract of Phrynium pubinerve Blume (EPP) supplement in attenuating allergic inflammation, encouraged by the presence of syringic acid in it, as this phenolic acid is reportedly promising in suppressing serum immunoglobulin E (IgE) and inflammatory cytokine levels. Materials and methodsHPLC-DAD dereplication analysis was performed to determine the presence of the vital polyphenolic metabolites. The efficacy of EPP against lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 cells was evaluated by measuring its inhibitory effects on NO and ROS/RNS production. The expressions of major inflammation-associated molecules (iNOS, COX-2, NF-κB, IL-6, and TNF-α) in RAW 264.7 cells were assessed through Western blot. Physiological and behavioral changes, BMI, and different biochemical parameters in mice blood serum were investigated in the toxicological assays. Formaldehyde-induced paw edema test in mice was conducted using established animal model. TDI-induced allergic model in mice was carried out to determine different allergy-like symptoms, and differential white blood cell (WBC) counts in blood and bronchoalveolar lavage (BAL) fluid. The intermolecular interaction analysis of the identified major metabolite of EPP with H1R and iNOS was studied by molecular docking. ResultsHPLC-DAD analysis showed the presence of syringic acid (89.19 mg/100 g EPP) and a few other compounds. LPS-induced NO generation was reduced by EPP in a concentration-dependent manner, showing IC50 of 28.20 ± 0.27 μg/mL. EPP exhibited a similar inhibitory effect on ROS/RNS production with IC50 of 29.47 ± 2.19 μg/mL. Western blotting revealed that EPP significantly downregulated the expressions of iNOS, COX-2, NF-κB, IL-6, and TNF-α in RAW 264.7 cells when challenged with LPS. The toxicological assays confirmed the dosage and organ-specific safety of EPP. In the formaldehyde-induced paw edema test, EPP caused a 66.41% reduction in mice paw volume at 500 mg/kg dose. It ameliorated TDI-induced allergy-like symptoms and decreased different inflammatory WBCs in mice's blood and BAL fluid in a dose-dependent manner. Finally, syringic acid demonstrated mentionable intermolecular binding affinity towards H1R (−6.6 Kcal/moL) and iNOS (−6.7 Kcal/moL). ConclusionsCollectively, considerable scientific reasoning was obtained in favor of the suppressive potential of EPP against allergic inflammatory responses that are proposed to be exerted via the downregulation of iNOS, COX-2, and NF-κB expressions, H1R antagonism and suppression of cytokines, such as IL-6, and TNF-α.

Investigation of the biological activity of the water-soluble C<sub>60</sub>/poly-<i>N</i>-vinylpyrrolidone complex

Objectives. The study aimed to investigate the biological activity of the C60/poly-N-vinylpyrrolidone (C60/PVP) complex representing a water-soluble fullerene derivative. In vitro and in vivo techniques were used to analyze the effect of the C60/PVP complex on the activity of lactate dehydrogenase (LDH) and evaluate changes in the biochemical parameters of blood serum when per os administered to mice.Methods. In order to determine the activity of a commercial LDH preparation and study the kinetics of this process, the standard Warburg photometric method was used. To assess the effect of polyvinylpyrrolidone (PVP) and the C60/PVP complex on some biochemical parameters in vivo, a study was conducted on two-month-old male white mongrel mice weighing 20 ± 3 g. Determination of biochemical parameters of blood serum was carried out using a semi-automatic biochemical analyzer according to standard methods.Results. The effect of the C60/PVP complex on LDH activity was studied along with changes in the biochemical parameters of mouse blood serum characterizing carbohydrate metabolism. As well as increasing the glucose and pyruvic acid content, the C60/PVP complex was found to reduce lactate content and LDH activity in blood serum along with in vitro LDH activity according to the type of mixed inhibition.Conclusions. The C60/PVP complex and PVP were shown to exhibit biological activity in vitro and in vivo. The C60/PVP complex, representing a mixed-type LDH inhibitor, was shown to inhibit LDH activity, as well as contributing to a decrease in lactate concentration and an increase in the concentration of pyruvic acid and glucose in blood serum when administered per os to mice. The inhibitory effect of PVP on LDH activity was revealed in both in vivo and in vitro investigations. In vivo, PVP contributes to a decrease in the concentration of lactate in the blood. The less pronounced effect of the C60/PVP complex as compared to PVP alone may be due to the fact that C60 molecules are “hidden” in cavities formed in PVP molecules.

Ag85A, As an S2 Vaccine Carrier, Reduces the Toxicity of the S2 Vaccine and Enhances the Protective Ability of Mice against Brucella.

Brucella is a globally distributed zoonotic disease that can cause abortion and changes in immune function in humans and animals. At present, there is no good treatment plan for Brucella, and animals can only be treated harmlessly once they become ill, resulting in huge economic losses. Therefore, the prevention of Brucella infection is a very crucial step. Although a variety of Brucella vaccines have been widely used, they have varying degrees of shortcomings. For example, some Brucella vaccines have residual virulence, which leads to the emergence of Brucella in animals during the immunization process. Bacillus infection and other conditions occur. To further reduce the toxicity of the Brucella vaccine and enhance its protective effect on animals, this study used Antigen 85A (Ag85A) as a carrier of the Brucella vaccine to fuse with the Brucella S2 vaccine. The results of the study found that the S2-Ag85A oral Brucella vaccine could effectively reduce the toxicity residue of the S2 vaccine, stimulate the mice to produce a better immunogenic response, and effectively activate the expression levels of Brucella heterozygous IgG1 and IgG2a. Experiments have shown that the expression of IFN-γ in the peripheral blood serum and spleen of mice is significantly increased, and the expression levels of IL-1β, TNF-α, and IL-6 are significantly reduced, which may indicate that S2-Ag85A oral Brucella vaccine could induce the expression of IFN-γ, thus downregulating the expression levels of IL-6 and TNF-α in the spleen tissue. The above results indicate that the S2-Ag85A oral vaccine is an effective attenuated vaccine for preventing Brucella infection.

Role of hypothalamic-pituitary-adrenal axis and catecholamine factors in independent and primary multiple types of tumor growth

Purpose of the study. To study the levels of adrenal axis factors in the hypothalamus, adrenal glands, blood serum of mice and catecholamines in the adrenal glands during the independent growth of B16 melanoma and Lewis lung carcinoma (LLC) and their combined growth in female mice, and in males – with independent growth of B16 and combined growth of B16 and LLC.Materials and methods. Male and female BALB/c Nude mice were divided into groups, n = 7 each: group 1 involved intact animals, group 2 involved mice with B16/F10 melanoma, group 3 – mice with LLC, group 4 – synchronous growth of melanoma and LLC. Levels of corticotropin releasing, noradrenaline and dopamine were determined in homogenates of the hypothalamus and adrenal glands and in the blood serum of all animals by ELISA, and levels of 17‑hydroxyprogesterone (17-OHP), dehydroepiandrosterone sulfate (DHEA-S) and cortisol were determined by RIA. Statistical processing of results was performed using the Statistica 10.0 program.Results. All tumor-bearing females showed elevated corticotropin releasing in the hypothalamus together with an increase of all stress-characterizing parameters: cortisol, the cortisol/DHEA-S ratio, and noradrenaline. However, an increase in serum levels of cortisol was blocked by high levels of DHEA-S, and as a result, the cortisol/DHEA-S ratio was either within the normal range (B16 melanoma and B16+LLC combination) or reduced (LLC). Levels of corticotropin releasing in the hypothalamus of tumor-bearing males decreased, together with opposite changes in stress-characterizing parameters in the adrenal glands: cortisol increased, the cortisol/DHEA-S ratio did not differ significantly from the control values, and noradrenaline decreased. An increase in serum levels of cortisol was not blocked by high levels of DHEA-S, and as a result, the cortisol/DHEA-S ratio was sharply elevated in B16 melanoma and B16+LLC combination.Conclusion. At independent and primary multiple types of tumor growth, the sex-specific features of the functioning of the adrenal axis at the central and peripheral levels are observed, which determines a more pronounced stressful state of the body with B16+LLC combination growth, realized by various mechanisms.

Analysis Protein APOB and TroponinT in Obese Mice (Musmusculus) Induced by Static Magnetic Field as a Marker of Coronary Heart

Obesity is closely related to cardiovascular disease. APOB protein was a better predictor of LDL particle count. Increased troponin T protein is also indicated as an indication of the risk of CHD disease. Static Magnetic Field (SMF) is an electromagnetic applied in therapy because only a simple magnetic disk is used to generate it both in vitro and in vivo. The use of SMF is considered to affect the biological state of cells so that it can affect weight loss in mice. The method used was male c57bl/6j mice with groups of Obes0, Obes2, Obes7, Obes 14, and Obes21 fed high-fat diet and then exposed to SMF with an intensity of 2mT for 1 hour. The results obtained were that there was a significant difference p=0.015 (p<0.05) in the protein concentration of APOB and there was no significant difference in the concentration of cTnT taken from the blood serum of mice. The increase in APOB protein levels in each control group was not found to have an increase in cTnT protein so that the two proteins did not have a linear correlation between the two proteins 0.598 (p>0.05). Most of the protein concentrations of APOB and cTnT decreased in concentration at longer days. It can be seen morphologically in the heart there is a decrease in the number of adipose cells.

Analysis of Mouse Blood Serum in the Dynamics of U87 Glioblastoma by Terahertz Spectroscopy and Machine Learning

In this research, an experimental U87 glioblastoma small animal model was studied. The association between glioblastoma stages and the spectral patterns of mouse blood serum measured in the terahertz range was analyzed by terahertz time-domain spectroscopy (THz-TDS) and machine learning. The THz spectra preprocessing included (i) smoothing using the Savitsky–Golay filter, (ii) outlier removing using isolation forest (IF), and (iii) Z-score normalization. The sequential informative feature-selection approach was developed using a combination of principal component analysis (PCA) and a support vector machine (SVM) model. The predictive data model was created using SVM with a linear kernel. This model was tested using k-fold cross-validation. Achieved prediction accuracy, sensitivity, specificity were over 90%. Also, a relation was established between tumor size and the THz spectral profile of blood serum samples. Thereby, the possibility of detecting glioma stages using blood serum spectral patterns in the terahertz range was demonstrated.

Free-standing vertically aligned tin disulfide nanosheets for ultrasensitive aptasensor design toward Alzheimer’s diagnosis applications

Despite a single drug, Aducanumab, recently approved by the U.S. Food and Drug Administration for Alzheimer’s disease (AD)-modifying treatments, the development of non-invasive, highly sensitive, and early screening devices to halt the disease remains a critical challenge. In this study, an electrochemical biosensor for amyloid-β (42) oligomers (AβOs) detection based on immobilized DNA aptamer onto vertical aligned tin disulfide nanosheets (SnS2 NSs) is developed for the first time. Highly crystalline, uniform, and vertical aligned SnS2 NSs were synthesized via a facile chemical vapor deposition (CVD) process. The sensing platform, comprising of Aβ (42)-specific capture aptamer covalently immobilized onto self-assembled-3-mercaptopropyl-trimethoxysilane (MPTMS)-SnS2 binds specifically toward AβOs. The biosensor can detect the AβOs at wide linear concentration range from 10-4 to 103 ng/mL with a low limit of detection (LOD) of 238.9 fg/mL and 56.9 fg/mL in physiological buffer and human serum (HS), respectively. The developed biosensor was successfully used to detect AβOs in the mice blood serum samples indicating its potential for clinical application in Alzheimer’s biomarker detection. Moreover, the SnS2-aptasensor shows good stability, selectivity, and interference-resistance. The predominant edge-plane active site and energetically preference for vertical SnS2 over its planar structure were unveiled by density-functional theory calculations which emphasize the advantages of applying nanomaterial with vertical orientated configuration into electrocatalytic reaction. The as-designed SnS2-aptasensor could shed light not only on multifunctional electrochemical biosensor design, but also for advancements on future Alzheimer’s diagnosis.

Neuroprotective and antioxidant activities of different polarity parts of the extracts of the Ginkgo biloba leaf and Zingiber officinale rhizome from Yongzhou.

In order to make better use of the two local dominant plant resources of Ginkgo biloba and Zingiber officinale from Yongzhou in Hunan province, the in vitro neuroprotective and antioxidant activities of extracts from the G. biloba leaf and Z. officinale rhizome, and the correlation between these two kinds of activities, were analyzed. The in vivo effects of these two plant extracts on aged mice blood physiology and central neuron cell activity were then determined after continuous gavage with the best polarity part at different concentrations (2, 4, 8 mg/ml). The results showed that the cell survival rate and superoxide dismutase (SOD) activity of the induced injury central neurons increased, although the malondialdehyde (MDA) content decreased gradually with the extract concentrations increasing in a certain range. Different polarity parts performed differently, even though they had the same concentration, with G. biloba always performing better than the Z. officinale rhizome at the same concentration and polarity. The order of G. biloba extract from superior to inferior was ethanol, ethyl acetate, n-butanol, chloroform, water, and petroleum ether (except that the petroleum ether part performed slightly better than the water part at 0.4 and 0.5 mg/ml), while the order of Z. officinale rhizome extract from superior to inferior was ethanol, chloroform, n-butanol, ethyl acetate, water, and petroleum ether. These two plant extracts demonstrated good in vitro effect against oxygen free radicals; the scavenging rate of superoxide free radicals had a significant positive correlation with the cell survival rate. The in vivo central nerve cell activity and SOD, glutathione peroxidase (GSH-PX) activity in aged mice blood serum increased while the MDA content decreased with treatment with these two extracts (p < 0.05). There were no significant changes in the number of leukocytes, lymphocytes, red blood cells, hemoglobin content, blood urine nitrogen, uric acid, creatinine, and the enzyme activity of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) (p > 0.05). G. biloba had a better in vivo effect than Z. officinale rhizome even though their concentration and polarity part were same. These results could provide some references for better development of these two plant extracts from Yongzhou in the field of neuroprotection.

RAC1 is involved in uterine myometrium contraction in the inflammation-associated preterm birth.

In briefVarious etiologies can cause uterine myometrium contraction, which leads to preterm birth. This study demonstrates a new functional relationship between the Ras-related C3 botulinum toxin substrate 1 (RAC1) and uterine myometrium contraction in preterm birth.Preterm birth (PTB) is a public health issue. The World Health Organization has recommended the use of tocolytic treatment to inhibit preterm labour and improve pregnancy outcomes. Intrauterine inflammation is associated with preterm birth. RAC1 can modulate inflammation in different experimental settings. In the current study, we explored whether RAC1 can modulate spontaneous uterine myometrium contraction in a mouse model of lipopolysaccharide (LPS)-induced intrauterine inflammation. Subsequently, we recorded uterine myometrium contraction and examined uterine Rac1 expression in a mouse model of preterm birth and a case in pregnant women by Western blotting analysis. We also measured progesterone levels in the blood serum of mice. Murine myometrium was obtained 12 h post LPS treatment. Human myometrium was obtained at the time of caesarean section. We found that in the LPS-treated group of mice, uterine myometrium contraction was enhanced, protein levels and activation of RAC1 were increased and serum progesterone levels were decreased. The protein levels of RAC1 were also increased in preterm birth and in pregnant women. NSC23766, a RAC1 inhibitor, attenuated uterine myometrium contraction and diminished RAC1 activation and COX-2 expression. Furthermore, silencing of RAC1 suppressed cell contraction and COX-2 expression in vitro. In conclusion, our results suggested that RAC1 may play an important role in modulating uterine myometrium contraction. Consequently, intervening with RAC1 represents a novel strategy for the treatment of preterm birth.

A Novel Method for Detecting Duchenne Muscular Dystrophy in Blood Serum of mdx Mice.

Duchenne muscular dystrophy (DMD) is the most common form of muscular dystrophy, typically affecting males in infancy. The disease causes progressive weakness and atrophy of skeletal muscles, with approximately 20,000 new cases diagnosed yearly. Currently, methods for diagnosing DMD are invasive, laborious, and unable to make accurate early detections. While there is no cure for DMD, there are limited treatments available for managing symptoms. As such, there is a crucial unmet need to develop a simple and non-invasive method for accurately detecting DMD as early as possible. Raman spectroscopy with chemometric analysis is shown to have the potential to fill this diagnostic need.

Arachidonic acid supplementation attenuates adipocyte inflammation but not adiposity in high fat diet induced obese mice

Obesity is associated with low-grade chronic inflammation and has a remarkable role in the pathophysiology of metabolic complications. In triggering these inflammatory responses, the arachidonic acid (AA) cascade plays a key role. However, there is a lack of data on how supplementary AA would affect obesity, adipose tissue inflammation, and the AA cascade in obesity. This study aims to investigate how AA supplementation affects obesity, adipocyte morphology, inflammation, and AA cascade signaling. Male Swiss Albino mice were used in our experiment. The mice were fed high-fat diets to induce obesity, and these obese mice were treated with two different doses of AA for 3 weeks. A normal diet non-obese group and an untreated obese group were kept as controls. Bodyweight and daily food intake data were recorded during that period. After the treatment period, blood serum and white adipose tissue of the experimental mice were collected for colorimetric lipid profile tests, histology, and mRNA extraction. The ΔΔCT method was employed for calculating the relative mRNA expression of target genes. The findings of our study suggest that AA has no significant effects on body weight, visceral adiposity, adipose tissue morphology, and serum lipid profile. However, AA treatment has resulted in a significant down-regulation of pro-inflammatory markers as well as the COX pathway. Besides, up-regulation of 12/15-LOX has been observed, indicating the metabolism pathway of supplementary AA through the LOX pathway. Our findings indicate that AA treatment may not provide significant benefits in terms of body weight, visceral fat mass, or serum lipid profile. However, it has effectively alleviated obesity-induced adipocyte inflammation in high-fat diet-induced obese mice.

Application of the method of dielectric spectroscopy to study theproperties of blood serum of mice with malignant ascites

The article presents the results of a study by the method ofdielectric spectroscopy of high-frequency relaxation processes in the bloodserum of intact mice and mice vaccinated with an oncological disease --- Ehrlich's ascites carcinoma. Using the formalism of the electrical module,the relaxation parameters were calculated for the serum samples of the twostudied systems. Keywords: electrical module, relaxation parameters, oncological disease.\

Применение метода диэлектрической спектроскопии для исследования свойств сыворотки крови мышей со злокачественным асцитом

The article presents the results of a study by the method of dielectric spectroscopy of high-frequency relaxation processes in the blood serum of intact mice and mice vaccinated with an oncological disease - Ehrlich's ascites carcinoma. Using the formalism of the electrical module, the relaxation parameters were calculated for the serum samples of the two studied systems.

Effect of Virgin Coconut Oil from Green Coconut on High-Density Lipoprotein (HDL) Levels in Blood Serum of White Mice

This study aims to determine the effect of High-Density Lipoprotein (VCO) dose on mice's blood serum High-Density Lipoprotein (HDL) levels. This study is an experimental study using 24 mice divided into four groups. Control (-), Control (+) were fed egg yolks 0.025 mL/g body weight, Dose 0.02 and Dose 0.025 were experimental groups, given egg yolks 0.025 mL/g mice body weight and VCO at a dose of 0.020 mL, and 0.025 mL/g body weight of mice. HDL levels were determined on days 7, 14, and 21 by enzymatic method using 20 D* spectrophotometry. The statistical test found that the length of time giving VCO significantly affected blood serum HDL levels in mice with Fcount &gt; Ftable (Fcount = 7.128 and Ftable = 5.14). In contrast, variations in VCO dose did not significantly affect blood serum HDL levels. mice with Fcount &lt; Ftable (Fcount = 3.33 and Ftable = 4.76). The highest HDL level was at a dose of 0.020 mL/g body weight of mice with a treatment period of 14 days, namely 164,033 mg/dL of blood.

Cholesterol Regulates Exosome Release in Cultured Astrocytes.

Exosomes are vesicles secreted by various kinds of cells, and they are rich in cholesterol, sphingomyelin (SM), phosphatidylcholine, and phosphatidylserine. Although cellular sphingolipid-mediated exosome release has been reported, the involvement of other lipid components of cell membranes in the regulation of exosome release is poorly understood. Here, we show that the level of exosome release into conditioned media is significantly reduced in cultured astrocytes prepared from apolipoprotein E (ApoE) knock-out mice when compared to those prepared from wild-type (WT) mice. The reduced level of exosome release was accompanied by elevated levels of cellular cholesterol. The addition of cholesterol to WT astrocytes significantly increased the cellular cholesterol levels and reduced exosome release. PI3K/Akt phosphorylation was enhanced in ApoE-deficient and cholesterol-treated WT astrocytes. In contrast, the depletion of cholesterol in ApoE-deficient astrocytes due to treatment with β-cyclodextrin recovered the exosome release level to a level similar to that in WT astrocytes. In addition, the reduced levels of exosome release due to the addition of cholesterol recovered to the control levels after treatment with a PI3K inhibitor (LY294002). The cholesterol-dependent regulation of exosome release was also confirmed by in vivo experiments; that is, exosome levels were significantly reduced in the CSF and blood serum of WT mice that were fed a high-fat diet and had increased cholesterol levels when compared to those in WT mice that were fed a normal diet. These results suggest that exosome release is regulated by cellular cholesterol via stimulation of the PI3K/Akt signal pathway.

Effect of Tilorone on the Dynamics of Viral Load and the Levels of Interferons and Interleukin-1β in the Lung Tissue and Blood Serum of Mice with Experimental Influenza.

We studied the effect of tilorone on the dynamics of IFNα, IFNγ, and IL-1β levels in the lung tissue and blood serum in relation to viral load in the lungs of BALB/c mice with pneumonia caused by influenza virus A/Aichi/2/68 (H3N2). Tilorone was administered per os in doses of 40, 150, and 540 μg per mouse 6, 30, and 78 h postinfection, which simulated the drug regimen used in the clinic for the treatment of influenza and acute respiratory viral infections in Russia and post-Soviet countries. Tilorone reduced viral load with the maximum amplitude (2-3 lg) after 1-2 administrations. The results of studying the dynamics of the cytokine levels in the infected animals in general support the previous hypothesis that, in repeated dosing, tilorone enhances the IFN response (compensates for its deficiency) at the early stages of acute respiratory viral infections and suppresses (damps) excessive production of IFN and proinflammatory cytokines at the later stages.

Overexpression of Hepatocyte Growth Factor in Dental Pulp Stem Cells Ameliorates the Severity of Psoriasis by Reducing Inflammatory Responses.

Psoriasis is an autoimmune disease still lacking standard treatment, and it has been demonstrated that mesenchymal stem cells (MSCs) are capable of immunoregulation. The underlying mechanism might involve the secretion of soluble cytokines, such as hepatocyte growth factor (HGF). This study aims to investigate the therapeutic effect of HGF-overexpressed dental pulp stem cells (DPSCs) [DPSCs; HGF overexpressed DPSCs (HGF-DPSCs)] on imiquimod-induced psoriasis. DPSCs were isolated and transfected by adenovirus vector carrying HGF gene (Ad-HGF). The immunoregulatry abilities of DPSCs and HGF-DPSCs were investigated by coculture of the MSCs with peripheral blood mononuclear cells (PBMCs) under appropriated stimulation. The psoriatic mice were treated with saline control, DPSCs, or HGF-DPSCs. Then the mice spleens were collected and weighted. The psoriatic skin lesions were analyzed by Hematoxylin/Eosin and immunohistochemical staining for histopathological changes, and quantitative real-time polymerase chain reaction to detect the expression levels of CD4+ T cell-related transcription factors and cytokines. The mice blood serum was measured by MILLIPLEX analysis and enzyme-linked immunosorbent assay to evaluate the expression levels of inflammation cytokines. The coculture experiments showed HGF overexpression enhanced the immunoregulation abilities of DPSCs not by suppressing PBMCs' proliferation, but by downregulating T helper 1 (Th1), Th17 cells, and upregulating regulatory T (Treg) cells. In psoriatic skin lesions, the psoriasis-like erythema, scaling, and thickening were ameliorated; and the expression of cytokeratin 6 (CK6), and cytokeratin 17 (CK17) were downregulated by DPSCs and HGF-DPSCs treatment. HGF overexpression enhanced the decrease of spleen masses; enhanced the downregulation of the expression levels of interferon-gamma (IFN-γ), tumor necrosis factor-α, and interleukin (IL)-17A in the blood serums; enhanced the downregulation of T-box transcription factor 21 (T-bet), IFN-γ, retinoic acid-related orphan receptor-γt (RORγt), IL-17A, IL-17F, IL-23, and upregulation of Foxp3 and IL-10 in the psoriatic skin lesions. Therefore, HGF overexpression enhanced DPSCs' treatment effect on psoriasis mainly by reducing inflammatory responses. These findings might provide new immunoregulation strategies for psoriasis treatment.

Abstract 314: Highly polymeric grape seed proanthocyanidins: A call for establishing the safe dose

Abstract Introduction: Proanthocyanidins (PC) are the oligomers and polymers of flavan-3-ols, especially catechin and epicatechin. PC in general are proven to be beneficial against cancers and widely available as over the counter (OTC) supplements. However, role of highly polymeric PC (HP-PC) on human cancer prevention remain unknown. Therefore, the effects of HP-PC supplementation on hepatic pathology in the context of hepatocellular carcinoma (HCC) were evaluated. HP-PC supplementation on mice gut microbiota was also evaluated considering the association of dysbiosis-mediated inflammation with HCC. Methodology: Crude PC was extracted from grape seeds (GS) using 70% acetone (in water) as the extraction solvent. PC mixture was purified and fractionated by flash chromatography. Monomeric and oligomeric PC was eluted with 100% ethanol and the HP-PC was collected by eluting with 70% acetone. C57BL/6 mice (49 - 56 days old) were supplemented with HP-PC (200 mg/kg body weight per day) for 14 days and sacrificed to collect livers and blood serum. Feces samples were collected from mice colon to evaluate the composition of gut microbiota using the 16S rRNA sequencing technology. Mice liver histopathology was examined by staining with hematoxylin and eosin. The level of alanine aminotransferase (ALT) in mice blood serum was measured as a marker of liver damage. Results: Evaluation of liver histopathology revealed signs of steatosis in the mice supplemented with HP-PC. Interestingly, rare foci of monocytic lobular inflammation was visible in the hepatic tissues of the mice supplemented with HP-PC. The alpha diversity of the mice gut microbiome was evaluated in terms of Shannon Index, Pielou's evenness, Faith's phylogenic diversity, and observed operational taxonomic units. A significant difference was not observed for the alpha diversity between the HP-PC supplemented and control group mice gut microbiota. The levels of ALT in the mice blood serum were not significantly different between HP-PC supplemented and control group mice. Conclusions: The preliminary experimental data suggests the potential of HP-PC to promote steatosis and hepatic inflammation. However, further studies must be conducted to understand potential harmful effect of HP-PC (if any) in hepatic toxicity and safer dose to use for potential health benefits. Furthermore, the results of this study emphasize the critical importance of establishing the safe dose of HP-PC or any OTC supplements. Citation Format: Wasitha Thilakarathna, H.P. Vasantha Rupasinghe. Highly polymeric grape seed proanthocyanidins: A call for establishing the safe dose [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 314.

Snail mucus suppresses anti dsDNA antibodies in lupus mice

Lupus is closely related to weather changes. Changes in extreme weather due to climate changes can trigger lupus flares and cause death. Therefore, many studies are trying to find a drug that can cure lupus. Snail mucus has the properties of suppressing the immune system. The aim of this study is to see how snail mucus affects the levels of anti-dsDNA antibodies in lupus model mice. Experimental study uses a posttest-only group design. The control group was mice with 0.5 cc intraperitoneal (IP) saline. The lupus nephritis group was mice with pristane 0.5 cc IP, and the therapy group was mice with 0.5 cc IP pristane and 0.5 cc snail mucus per day at the 4th month of treatment for 14 days and after that the mice blood serum was taken and examined in the laboratory. The dsDNA antibody examination used the ELISA method. Statistical test with Anova followed by post hoc test. P is significant if it is less than 0.05. There was an increase in the levels of antibody dsDNA (p = 0.016) in the lupus group (75.13 + 22.5mg/dL) compare with the control group (56.96 ± 11.77 mg/dL), but there was a decrease in anti-dsDNA antibodies in the mice with snail mucus (56.01 ± 8.29 mg/dL; p = 0.02), and the methyl prednisolone group (53.47 ± 13.53 mg/dL; p = 0.009) when compared to the lupus group. Snail mucus suppresses anti dsDNA antibody levels in lupus mice.

Serum exosomes accelerate diabetic wound healing by promoting angiogenesis and ECM formation.

Nonhealing wounds in diabetes remain a global clinical and research challenge. Exosomes are primary mediators of cell paracrine action, which are shown to promote tissue repair and regeneration. In this study, we investigated the effects of serum derived exosomes (Serum-Exos) on diabetic wound healing and its possible mechanisms. Serum-Exos were isolated from blood serum of normal healthy mice and identified by transmission electron microscopy and western blot. The effects of Serum-Exos on diabetic wound healing, fibroblast growth and migration, angiogenesis and extracellular matrix (ECM) formation were investigated. Our results showed that the isolated Serum-Exos exhibited a sphere-shaped morphology with a mean diameter at 150 nm, and expressed classical markers of exosomes including HSP70, TSG101, and CD63. Treatment with Serum-Exos elevated the percentage of wound closure and shortened the time of healing in diabetic mice. Mechanistically, Serum-Exos promoted granulation tissue formation and increased the expression of CD31, fibronectin and collagen-ɑ in diabetic mice. Serum-Exos also promoted the migration of NIH/3T3 cells, which was associated with increased expression levels of PCNA, Ki67, collagen-α and fibronectin. In addition, Serum-Exos enhanced tube formation in human umbilical vein endothelial cells and induced the expression of CD31 at both protein and messenger RNA levels. Collectively, our results suggest that Serum-Exos may facilitate the wound healing in diabetic mice by promoting angiogenesis and ECM formation, and show the potential application in treating diabetic wounds.