Serum Of Animals Research Articles

A Global Collaborative Comparison of SARS-CoV-2 Antigenicity Across 15 Laboratories

Setting up a global SARS-CoV-2 surveillance system requires an understanding of how virus isolation and propagation practices, use of animal or human sera, and different neutralisation assay platforms influence assessment of SARS-CoV-2 antigenicity. In this study, with the contribution of 15 independent laboratories across all WHO regions, we carried out a controlled analysis of neutralisation assay platforms using the first WHO International Standard for antibodies to SARS-CoV-2 variants of concern (source: NIBSC). Live virus isolates (source: WHO BioHub or individual labs) or spike plasmids (individual labs) for pseudovirus production were used to perform neutralisation assays using the same serum panels. When comparing fold drops, excellent data consistency was observed across the labs using common reagents, including between pseudovirus and live virus neutralisation assays (RMSD of data from mean fold drop was 0.59). Utilising a Bayesian model, geometric mean titres and assay titre magnitudes (offsets) can describe the data efficiently. Titre magnitudes were seen to vary largely even for labs within the same assay group. We have observed that overall, live Microneutralisation assays tend to have the lowest titres, whereas Pseudovirus Neutralisation have the highest (with a mean difference of 3.2 log2 units between the two). These findings are relevant for laboratory networks, such as the WHO Coronavirus Laboratory Network (CoViNet), that seek to support a global surveillance system for evolution and antigenic characterisation of variants to support monitoring of population immunity and vaccine composition policy.

Progress in the application of auto-concentrated growth factor (CGF) in wound repair.

Auto-concentrated growth factor (CGF) constitutes the latest generation of plasma extract, and has high concentrations of growth factors and white blood cells. Due to the continuous variable speed centrifugation used during preparation, the tensile strength of the fibrin is also higher. CGF preparation does not involve the use of animal serum, minimizing the risk of infection and immune rejection. Therefore, it has wide potential applications in various fields of regenerative medicine. This paper summarizes the history behind CGF development, reviews the clinical applications and research progress concerning single CGF therapy and CGF used in combination with other treatments in multiple wound repair, and summarizes its potential value as therapeutic agent. Finally, some constructive suggestions and research perspectives for the application of CGF in wound healing are put forward. The available evidence indicates that CGF can promote the healing of chronic refractory wounds and acute wound, promote the growth of granulation, accelerate the speed and improve the quality of wound healing, reduce scar formation, minimize the need for repeated wound dressing, and ameliorate the pain experienced by patients.

In vivo Study of the Anti/Prooxidant Activity of a Metal-Organic Coordination Polymer Modified with Iron Oxides and Ascorbic Acid

Experiments on 39 male Wistar rats were performed. A metal-organic coordination polymer modified with iron oxide and ascorbic acid (composite) was administered intravenously to healthy rats at doses of 25 and 50 mg/kg. The anti/prooxidant activity of the composite was assessed 3 and 24 hours after administration. The composite in both doses was shown to have no significant effect on the homeostasis of prooxidants/antioxidants in the blood serum in healthy animals. In high doses, the composite enhances apoptosis of lymphocytes, which indicates its possible use as a catalyst for free radical processes for further use in biomedicine.

The green formulation of silver nanoparticles for the anticancer and lupus nephritis treatment applications and reducing the inflammatory cytokines in the rat periodontal model

In this study, a novel bio-inspired synthesis for the preparation of Ag NPs by Calendula persica flower extract has been researched. Also, the effects of the recent composite on lupus nephritis by following the TGF-β1 signaling pathway in mice and gingival amounts of IL1-β and TNF-α in the animal periodontal model were determined. To assess the crystal nature, morphology and size of the prepared nanoparticles, X-ray diffraction (XRD), transmission electron microscopy (TEM), ultraviolet–visible (UV–Vis) and field emission-scanning electron microscopy (FE-SEM) analyses were applied in this study. The prepared Ag NPs was employed for its effectiveness against colon cancer cell line (C26). The bio-synthesized Ag NPs had also cytotoxic property versus colon cancer based on MTT protocol. Collectively, the Ag NPs fabricated by biological way has potential applications in inflammatory treatment. In lupus nephritis section, indexes in animal tissues and serum were identified using specific kits, while pathological alterations in the mice kidney were examined through H&E staining. Additionally, qPCR was employed to determine the expression related to TGF-β 1 in kidney, thereby providing further insight into the silver nanoparticles mechanism. The findings indicated that silver nanoparticles reduced the mRNA expression levels of TGF-β 1 and NF-κB in the kidneys, while simultaneously increasing the Mn-SOD and IκB-α expression. Histological examination of H&E-stained sections revealed that silver nanoparticles mitigated the morphological abnormalities of the glomeruli and the inflammatory infiltration associated with nephritis. Silver nanoparticles demonstrated an inhibitory effect on the dsDNA positive rate in animal suffering from nephritis. Silver nanoparticles were found to reduce the increase in urinary protein and the pro-inflammatory cytokines concentrations in both the kidney and serum. In the periodontal model study, 0-3 ligatures were applied to induce the inflammatory periodontitis in right mandibular first molar neck in rats. The levels of inflammatory cytokines (IL1-β, CINC-1, IL-10 and TNF-α) were determined by common immunological test, ELISA. The findings revealed that Ag NPs administration could decease the IL1-β and TNF-α production in the rat periodontal model gum tissue. In addition, the outcome of this research indicated that IL1-β and TNF-α levels raised in the rat periodontal model gum tissue compared to control group. Ag NPs increased the levels of superoxide dismutase, catalase, and plasma bone alkaline phosphatase and reduced the IL1-β, TNF-α, inducible nitric oxide synthase and RANK genes mRNA expression. The results indicate that pre/post-treatment with Ag NPs because of its direct activity on pro-inflammatory cytokines inhibition improved the inflammatory symptoms in the periodontitis and lupus nephritis animals.

O USO DE SORO HIPERIMUNE NA CLÍNICA DE PEQUENOS ANIMAIS

The immune system is intended to defend the body against harmful invaders. Antibodies are largely responsible for defense against pathogenic antigens. The immunological defense of some conditions, both innate and acquired, may have nonspecific production and insufficient antibodies to combat them. The use of hyperimmune serum can be an alternative in defending the body against specific pathogens, a procedure which transfers antibodies ready for defense against antigens. This therapy consists of using plasma or serum extracted by centrifugation from the blood of an animal previously vaccinated or already cured of a specific disease. The serum is offered to an animal that has had a failure in the vaccination protocol or has contracted a certain disease. The objective of this study was to carry out a literature review regarding the use and application of hyperimmune serum in small animals. Also used as complementary therapy, hyperimmune serum has demonstrated excellent results in reducing free antigens in the serum of patients affected by certain pathologies. It proved to be relevant in clinically improving recovery from diseases such as distemper and parvovirus. This technique presents a promising prognosis, but there are many studies to be carried out to effectively prove the application of hyperimmune serum in the complementary treatment of certain diseases that affect dogs and cats.

Ameliorative Potency of Iridoid Glucoside Compound Catalpol on Inhibiting NF-κB-mediated Inflammation in Allergic Rhinitis-induced Mice

Background Over time, the prevalence of allergic rhinitis (AR) has surged due to various risk factors, notably attributed to global urbanization, rendering heightened levels of pollutants, including traffic-related emissions and particulate matter. About 25% of the global children population and 40% of the adult population were reported with AR. Even though AR is recognized as a systemic inflammatory disease, it often results in diverse other comorbidities, such as dermatitis, sinusitis, conjunctivitis, and otitis, requiring extensive and expensive treatment. Purpose We assessed the effectiveness of iridoid glucoside catalpol against AR in a mouse model. Catalpol is recommended in traditional Chinese medicine to treat diverse acute and chronic diseases. Methods AR was induced in mice with an ovalbumin-sensitized AR model and treated with 10 and 20 mg of catalpol. Nasal severity scoring was performed to confirm the AR induction in mice. Allergic mediators immunoglobulin E (IgE) and histamine were quantified in the serum to assess the anti-allergic response of catalpol. In nasal lavage fluid (NALF), the inflammatory mediators IgE Ab, prostaglandin D2, leukotriene C4, eosinophil cationic protein (ECP), and pro-inflammatory cytokines were measured to analyze the anti-inflammatory potency of catalpol. The binding capacity of nuclear factor-kappa B (NF-κB) to DNA was evaluated to assess the catalpol inhibitory potency against NF-κB-mediated inflammation in AR mice. To confirm the ameliorative potency of catalpol in AR mice, a histopathological analysis of nasal mucosa was performed. Results Catalpol treatment significantly decreased the nasal symptoms and reduced the allergic mediators in the serum of experimental animals. It effectively inhibited the synthesis of inflammatory mediators, ECP, and pro-inflammatory cytokines in the NALF, and also suppressed the NF-κB DNA-binding activity in AR mice. The decrease in ciliary loss, goblet cells, eosinophil infiltration, and vascular congestion observed with our nasal mucosa histopathological analysis confirmed the ameliorative potency of catalpol. Conclusion Our findings have proven catalpol inhibits NF-κB-mediated inflammatory response in AR mice. With further analysis, a potent natural compound, catalpol, can be formulated as a drug to treat AR.

The effect of exogenous melatonin on the dynamics of body weight and the level of Klotho protein in the blood of female rats subjected to prolonged dark deprivation

The study of the effect of exogenous melatonin (0.3 mg/kg, intragastrically in 2% starch mucus, 14-day course) on the dynamics of body weight and the level of Klotho protein in the blood of 4-month-old white mongrel female rats subjected to 30-day dark deprivation (light-dark mode 24/0 h, artificial lighting 300 Lux). It has been shown that prolonged dark deprivation leads to a decrease in the level of Klotho protein, which is a marker of premature aging; a decrease in melatonin levels, fluctuations in the rate of body weight gain in sexually mature female rats. Exogenous melatonin contributed to an increase in the level of Klotho protein and the restoration of melatonin levels in the blood serum of animals, as well as a decrease in body weight, which indicates its protective effect on the condition of animals with accelerated aging caused by dark deprivation.

Interferon as an immunoadjuvant to enhance antibodies following influenza B infection and vaccination in ferrets

Despite annual vaccination, influenza B viruses (IBV) continue to cause significant morbidity and mortality in humans. We have found that IBV infection resulted in a weaker innate and adaptive immune response than influenza A viruses (IAV) in ferrets. To understand and overcome the weak immune responses to IBV in ferrets, we administered type-I or type-III interferon (IFN) to ferrets following infection or vaccination and evaluated their effects on the immune response. IFN signaling following viral infection plays an important role in the initial innate immune response and affects subsequent adaptive immune responses. In the respiratory tract, IFN lambda (IFNL) has regulatory effects on adaptive immunity indirectly through thymic stromal lymphopoietin (TSLP), which then acts on immune cells to stimulate the adaptive response. Following IBV infection or vaccination, IFN treatment (IFN-Tx) upregulated gene expression of early inflammatory responses in the upper respiratory tract and robust IFN, TSLP, and inflammatory responses in peripheral blood cells. These responses were sustained following challenge or vaccination in IFN-Tx animals. Serum IFNL and TSLP levels were enhanced in IFN-Tx animals following challenge/rechallenge over mock-Tx; however, this difference was not observed following vaccination. Antibody responses in serum of IFN-Tx animals following IBV infection or vaccination increased more quickly and to higher titers and were sustained longer than mock-Tx animals over 3 months. Following rechallenge of infected animals 3 months post treatment, antibody levels remained higher than mock-Tx. However, IFN-Tx did not have an effect on antibody responses following challenge of vaccinated animals. A strong direct correlation was found between TSLP levels and antibody responses following challenge-rechallenge and vaccination-challenge indicating it as a useful tool for predicting adaptive immune responses following IBV infection or vaccination. The effects of IFN on strengthening both innate and adaptive responses to IBV may aid in development of more effective treatments following infection and improved influenza vaccines.

Antioxidant and anti-glycation activities of Mandevilla velutina extract and effect on parasitemia levels in Trypanosoma cruzi experimental infection: In vivo, in vitro and in silico approaches

Ethnopharmacological relevanceMandevilla velutina (Mart. Ex Stadelm.) Woodson, known in Brazil as "infalível" and "jalapa", is a medicinal plant native from the Cerrado region (Brazilian Savannah). The underground organ (xylopodium) of this species is prepared as ethanolic extract or infusion and it is commonly used in traditional medicine to treat snake venom. Although, locals and indigenous populations from Cerrado have used M. velutina for the treatment of infection by Trypanosoma cruzi (Chagas’ disease). Aim of the studyThis study aimed to evaluate the in vitro antioxidant and anti-glycation activities of the crude hydroethanolic extract of M. velutina xylopodium. Besides, it aimed to evaluate its effect on parasitemia levels in vivo T. cruzi experimental infection. In addition, this study aimed to determine possible interactions between the main compound of the extract and molecular targets associated with survival and virulence of T. cruzi in silico approaches. Materials and methodsDetermination of total polyphenols, flavonoids and steroidal aglycones content were performed. In addition, high performance liquid chromatography (HPLC) was carried out to identify main compounds of the extract. Antioxidant activity was determined by DPPH radical scavenging, ferric ion reducing power (FRAP), Thiobarbituric acid reactive species (TBARS) and Oxygen Radical Absorbance Capacity (ORAC) methods. Anti-glycation activity was demonstrated through relative mobility in electrophoresis (RME), determination of free amino groups and inhibition of AGEs formation. Determination of the action of extract in parasitemia levels was performed by T. cruzi experimental infection of mice and nitrite levels were measured in the serum of animals evaluated in this study. Molecular docking analyses of the main compound (Velutinol A) with DNA and molecular targets associated with survival and virulence of T. cruzi. ResultsPhytoconstituents evaluation exhibited the presence polyphenols, flavonoids and steroidal aglycone, and HPLC identified the major presence of Velutinol A. Antioxidant and anti-glycation evaluations showed that the extract present significant activity in all methods evaluated. In addition, extract reduced the number of trypomastigotes and increased the survival of treated animals. The treatment using extract showed an interference in the synthesis of physiological nitric oxide as an immune response to infection. In silico assays demonstrated interaction between Velutinol A and DNA and molecular targets of T. cruzi. ConclusionsThe results showed that the hydroethanolic extract of M. velutina xylopodium contains bioactive compounds including polyphenols, flavonoids and steroidal aglycones (mainly Velutinol A) of which may be responsible for the antioxidant, anti-glycation and anti-parasitic activity against T. cruzi. Trypanocidal activity of M. velutina compounds may be linked to their influence on NO synthesis during infection and/or their capacity to bind and inhibit molecules associated to virulence and survival of T. cruzi.

Hypoazotemic action of dry extracts from the waste of Tribulus terrestris

5 samples of dry extract were obtained from the processing waste of the aerial part of Tribulus terrestris L., from the meal after extraction with 70% ethanol. Based on pharmacological studies, it was established that dry extract from Tribulus terrestris meal, containing 22% polysaccharides and 31.3% of the total phenolic compounds, exhibits a hypoazotemic effect, which has a positive effect on kidney function. In the experimental model of acute kidney failure caused by the introduction of a 50% aqueous solution of glycerol and acute kidney pathology caused by iodine tincture, Triburenal substance obtained from Tribulus terrestris meal has a significant hypoazotemic action. This prevents a sharp increase in the end products of nitrogen metabolism in the blood serum of animals and reduces these indicators more than the reference drug Cynaroside. The urea content in the blood serum of rats receiving Triburenal on the third day of pathology, the difference from the control was 52.0%. Similar changes were noted in the creatinine content in the blood serum; the difference with the control was 49.0%. Regarding the degree of reduction of urea and creatinine in the blood, the Triburenal substance from Tribulus terrestris meal is superior to the hypoazotemic drug Cynaroside. A technological scheme has been proposed for the production of a pharmaceutical substance in the form of a dry extract with a hypoazotemic action from Tribulus terrestris meal with a final product yield of 5.65% by weight of the raw material. The technology is as follows: after extraction of the aerial part of Tribulus terrestris with 70% ethanol, the meal is dried in a drying apparatus with forced air ventilation with a layer of plant raw material on a drying baking tray of 20 mm thick, supplying air at a temperature of 70°C at a speed of 15 m/s for 5 hours, the dried meal is extracted with 40% ethanol at a temperature of 60°C four times at a hydro modulus of 1:18, infusing for 3 hours, the combined extract is concentrated to a content of 15% dry weight and dried. The results obtained make it possible to use the aerial part of Tribulus terrestris rationally.

Detection of replication-competent circular single stranded DNA from Deoni cattle

Bovine meat and milk factors (BMMF) having structural similarity to plasmids of Acinetobacter baumanni and some single stranded (ss) DNA viruses are free circulating replication-competent circular ssDNA molecules which were first isolated from serum of Bos taurus animals. Isolation of BMMF from tissues of multiple sclerosis as well as some cancers led researchers to conclude that the consumption of these agents through milk and meat have a positive correlation with these diseases and that the absence of such molecules in B. indicus animals could account to the low occurrence of cancer in those areas consuming products of indigenous animals. Hence, the present study was designed to investigate its presence in Deoni, one of the indigenous cattle breeds of India. The DNA isolated from milk samples were subjected to rolling circle amplification (RCA) using exo-resistant random primers. The products of RCA were then used as template for PCR for amplification of circular DNA. The amplicons obtained of size around 2000bp were cloned in E. coli and sequenced. The BLASTn, multiple sequence alignment and phylogenetic analysis of the obtained sequences were carried out to identify the similarities of these isolates. The sequence showed maximum homology with either BMMF2 or plasmids of Acinetobactor spp. Replication initiation protein (Rep) which showed similarity to bacterial plasmids was present in the sequence. This is the pioneer report on the presence of BMMF in milk of indigenous cattle of India. Keywords: BMMF, replication-competent, circular ssDNA, Deoni

Lifetime minimally invasive assessment of iron in the pig liver

Microelements are an integral part of the mammalian body, and their content in organs and tissues is associated with other components of a complex biological system. Based on this, it is feasible to evaluate the concentration of specific chemical elements within the structures of the body in non-invasive or minimally invasive methods. The meat and by-products of farm animals serve as a readily assimilateable source of iron, which is one of the reasons for potentially defining the quality of agricultural products in conditions of widespread iron deficiency. Landrace pigs were raised in standard conditions at an industrial complex located in the Altai Territory in order to fatten up to a live weight of 100 kg. Venous blood was collected using the acute method from the jugular vein in accordance with the principles of asepsis and pre-analytical guidelines. The hematologic and biochemical examination of the blood and serum of animals was performed by apparatus. After slaughter, liver samples were collected, and the method of atomic emission spectral analysis using inductively coupled plasma on iCAP-PRO equipment (Thermo Fisher Scientific) was used to estimate the iron level in them. To manipulate the data, Microsoft Office Excel software and RStudio data analysis environment version 2023.03.1 (RStudio, PBC) were employed. For regression analysis, the least squares approach was used. The model was fitted using a stepwise selection of predictors in both directions using the Akaike information criterion, Bayesian information criterion, and adjusted coefficient of determination. The linear regression assumptions were evaluated. The final regression model used for determining iron levels in pig liver contains mean hemoglobin content in erythrocytes, hemoglobin, and serum inorganic phosphorus as predictors. There is no evidence that there is multicollinearity between the predictors of the final model. The proposed model satisfies the requirements for a normal distribution of residuals, the absence of their correlation, and influential observations. The proposed multiple regression model has the capability to estimate iron levels in pig liver in vivo for various purposes.

Ultrasensitive and highly selective Co2+ detection based on the chiral optical activities of L-glutathione-modified gold nanoclusters.

Developing highly sensitive and selective detection methods is crucial for environmental and healthcare monitoring. In this study, the chiral and fluorescent signals of L-glutathione-modified gold nanoclusters (L-GSH-Au NCs) were discovered to be responsive to Co2+, which displayed linear correlations with the concentration changes of Co2+. Notably, the chiral signal was more sensitive than the FL signal, whose limit of detection (LOD) was calculated to be 0.37μM and 3.93 times lower than the LOD obtained with fluorescent signals. Moreover, the chiral signals exhibited unexpectedly high selectivity towards Co2+, effectively avoiding interference from other metal ions and biomolecules. Furthermore, the concentrations of Co2+ in various samples, such as Taihu water, tap water, bottled water, and animal serum, were accurately quantified using the chiral signals of L-GSH-Au NCs without complex pretreatment, with recoveries ranging between 95.64% and 103.22%. This study not only provides an innovative approach for Co2+ detection but also highlights the detection capabilities of chiral signals in complex environments.

Mesoporous Silica Administration as a New Strategy in the Management of Warfarin Toxicity; In Vitro and In Vivo Study

Purpose: Warfarin is one of the most widely used anticoagulants that function by inhibiting vitamin K epoxide reductase. Warfarin overdose, whether intentional or unintentional, can cause life-threatening bleeding. Here, we present a novel warfarin adsorbent based on mesoporous silica that can act as an antidote to warfarin toxicity. Method: Amino-functionalized mesoporous silica (MS-NH2) was synthesized based on co-condensation method through a soft template technique followed by template removal. The prepared structure and functional group were studied by FT-IR, XRD. The morphology was checked by SEM and TEM. The capacity of MS-NH2 in the adsorption of warfarin was evaluated in vitro, at pH=7.4 and pH=1.2. In vivo evaluation was performed in control and warfarin-overdosed animal models. Overdosed animals were treated with MS-NH2 by oral gavage. Biomarkers of organ injury were assessed in animal serum. Results: The MS-NH2 were relatively uniform, spherical with defined diameters (400 nm) and homogeneous structure. synthesized particles had a large surface area (1015 m2 g-1) and mean pore diameter 2.4 nm which leaded considerable adsorption capacity for warfarin 1666 mg/g. In vivo studies revealed that oral administration of MS-NH2 in mice poisoned with warfarin caused a significant difference (p < 0.05) on International Normalized Ratio (INR) and prothrombin time (PT). Moreover, the warfarin with MS-NH2 group demonstrated a notable decrease in biomarkers associated with tissue damage, such as bilirubin, lactate dehydrogenase (LDH), alanine aminotransferase (ALT), and aspartate aminotransferase (AST). Conclusion: The results confirm that MS-NH2 administration can be an effective treatment for warfarin toxicity and could potentially mitigate the adverse effects of warfarin poisoning.

B-246 Pab to mab conversion via mass spectrometry: direct antibody discovery from serum

Abstract Background Navigating the intricacies of discovering unique and functional antibody binders for in vitro diagnostic development comes with several technological challenges, such as limited diversity of antibody repertoires, functional screening challenges, and the absence of suitable in vitro models. A proteomics-driven strategy to antibody discovery is a promising solution to overcome these obstacles by facilitating polyclonal to monoclonal antibody conversion. De novo polyclonal antibody sequencing utilizes mass spectrometry and machine-learning driven bioinformatics to obtain complete antibody sequences from immunoserum or purified protein samples. This approach investigates the entire circulating antibody repertoire at the protein level, thereby broadening the spectrum of potential candidates. It further facilitates functional selection by enabling antibody purification and enrichment techniques. This capability facilitates the identification of antibodies possessing unique functions, crucial for subsequent downstream development. We demonstrate the application and de novo polyclonal antibody sequencing using mass spectrometry and machine learning bioinformatics to enable the next generation of antibody discovery. Methods In this study, we employed a novel mass spectrometry-based approach, REpAb, to de novo sequence polyclonal antibodies directly from the immunized serum of animals or human patients. REpAb integrates mass spectrometry with machine learning-based bioinformatics to de novo sequence and assemble monoclonal antibodies from a polyclonal mixture. Protein lysates underwent analysis using an Orbitrap EclipseTM Series instrument (ThermoFisher Scientific, CA, US) coupled with the LC Evosep One (Evosep, Denmark). Results De novo polyclonal sequencing with REpAb yielded unique antibody binders not found in splenocytes or peripheral blood mononuclear cells (PBMCs). De novo antibody sequencing-discovered antibody sequences from naturally exposed humans showed unrestricted germline usage, compared to restricted germline used in antigen-immunized animals. Additionally, a proteomics-based approach for antibody discovery enables functional antibody selection through standard protein A/G purification, followed by negative and positive selection enrichment strategies to isolate antibody clones specific to the region of interest on the antigen. Conclusions Sequencing antibodies at the protein level directly reflects the circulating antibody repertoires, which expands the antibody diversity from the BCR repertoire. Proteomic-based antibody discovery empowers the functional selection of antibodies via meticulously designed enrichment strategies. This approach allows for the identification of a diverse pool of candidates possessing specific functions, such as anti-idiotypic features.

Tectorigenin Inhibits Glycolysis-induced Cell Growth and Proliferation by Modulating LncRNA CCAT2/miR-145 Pathway in Colorectal Cancer.

Colorectal cancer (CRC) places a heavy burden on global health. Tectorigenin (Tec) is a type of flavonoid-based compound obtained from the Chinese medical herb Leopard Lily Rhizome. It was found to exhibit remarkable anti-tumor properties in previous studies. However, the effect and molecular mechanisms of Tec in colorectal cancer have not been reported. The objective of this study was to explore the action of Tec in proliferation and glycolysis in CRC and the potential mechanism with regard to the long non-coding RNA (lncRNA) CCAT2/micro RNA-145(miR-145) pathway in vitro and in vivo . The anti-tumor effect of Tec in CRC was examined in cell and animal studies, applying Cell Counting Kit-8 (CCK-8) assay as well as xenograft model experiments. Assay kits were utilized to detect glucose consumption and lactate production in the supernatant of cells and animal serum. The expression of the glycolysis-related proteins was assessed by Western Blotting, and levels of lncRNA CCAT2 and miR-145 in CRC tissue specimens and cells were assessed by realtime quantitative PCR (RT-qPCR). Tec significantly suppressed cell glycolysis and proliferative rate in CRC cells. It could decrease lncRNA CCAT2 in CRC cells but increase the expression of miR-145. LncRNA CCAT2 overexpression or inhibition of miR-145 could abolish the inhibitive effects of Tec on the proliferation and glycolysis of CRC cells. The miR-145 mimic rescued the increased cell viability and glycolysis levels caused by lncRNA CCAT2 overexpression. Tec significantly inhibited the growth and glycolysis of CRC xenograft tumor. The expression of lncRNA CCAT2 decreased while the expression of miR-145 increased after Tec treatment in vivo. Tec can inhibit the proliferation and glycolysis of CRC cells through the lncRNA CCAT2/miR-145 axis. Altogether, the potential targets discovered in this research are of great significance for CRC treatment and new drug development.

THE IMPACT OF HOP EXTRACT ACTIVE COMPONENTS ON METABOLIC DISTURBANCES IN ESTROGEN DEFICIENT FEMALE RATS WITH PREDIABETES

A number of studies have shown a greater cardiovascular risk in women with type 2 diabetes compared to men. The latter may be due to more excessive manifestations of metabolic disorders in women, in particular postmenopausal, that develop even at the stage of prediabetes. Since the use of hormone replacement therapy in women after menopause had adverse results on the progression of cardiovascular events, an alternative method of reducing cardiovascular risk in postmenopausal women with prediabetes may be the use of phytoestrogens, which possess estrogen-like activity without negative side effects specific to exogenous estrogens. The aim of our study was to evaluate the effect of hop extract on metabolic disturbances in female rats with prediabetes under estrogen deficiency. Materials and methods. The study was conducted on three-month-old intact and ovariectomized Wistar rats with metabolic syndrome (MS) induced by high calorie diet (15% lard, 25% sucrose, 1% bile salts, 59% standard feed) for eight weeks. The test substances were administered intragastrically once per day in a dose of 20 μg/kg or 200 μg/kg b.w. in recalculation on 8-prenylnaringenin for hop extract and 200 μg/kg b.w. in recalculation on estradiol for the reference drug, from the fifth week after induction of metabolic syndrome. Results. It was revealed that administration of hop extract in a dose of 200 μg/kg, similar to estradiol, led to improvement in glucose tolerance and decrease in insulin resistance (p˂0.05) in estrogen deficient rats with MS. In contrast to estradiol, hop extract had no effect on the mass of uterus in ovariectomized animals. The use of hop extract only in the dose of 200 μg/kg, similar to estradiol, was accompanied by a significant decrease in body weight gain and visceral fat, as well as normalization of the total lipids content in serum of experimental animals (p˂0.05). It is of note, that hop extract resulted in decrease of hypertriglyceridemia (p˂0.05), but did not affect the level of total cholesterol, while estradiol reduced the concentration of total cholesterol (p˂0.05), and had no effect on the level of triglycerides. Hop extract in a dose of 200 μg/kg, as well as estradiol, led to a decrease in the intensity of lipid peroxidation induced by MS under estrogen deficiency, as evidenced by reduced concentration of conjugated dienes in serum (p˂0.05). At the same time, the total antioxidant activity of serum in these animals remained elevated compared to intact group. In rats with MS and hypoestrogenia, hop extract in the highest dose and estradiol resulted in complete normalization (p˂0.05) of the decreased level of NO stable metabolites in serum and the reduced activity of NO-synthase in heart. Conclusions. The revealed cardioprotective effect of hop extract that is realized due to the improved lipid profile and reduced insulin resistance, visceral obesity, oxidative stress and endothelial dysfunction in estrogen deficient female rats with MS indicates the prospects of its use for prevention and treatment of cardiovascular complications in postmenopausal women with prediabetes.

A nanohybrid-based smartphone-compatible high performance electrochemical glucose sensor

A cost-effective and low-potential smartphone-compatible electrochemical sensor was constructed using a nanohybrid for the sensing of glucose in human and animal serum. The nanohybrid composed of gold nanoparticle (GNP) decorated cobalt hexacyanoferrate (CHCF) modified ZIF-67 (cobalt metal organic framework, CMOF) was synthesized and characterized by FTIR, XRD, and SEM with EDX analysis. The electrochemical characteristics of the nanohybrid were investigated by depositing the nanohybrid over a conventional glassy carbon electrode (GCE) and performing cyclic voltammetry, which revealed a stable redox peak with a formal potential of +0.23 V, corresponding to Co2+/3+ redox couple in GNP–CHCF–CMOF. Thus, developed sensor was utilized for the electrochemical glucose detection, which showed exceptional electrocatalytic activity over a linear detection range from 8.33 to 3793 μM with a low detection limit of 0.96 μM at a low potential of +0.35 V. Furthermore, the GNP–CHCF–CMOF/GCE sensor was employed for the detection of glucose spiked in human and rabbit serum samples, which showed excellent recoveries. A portable measurement device was fabricated which showed the real-time monitoring of glucose in a smartphone. This novel approach paves the way for the design and fabrication of cost-effective, low-potential sensors, which would reduce overall costs and enhance the performance of sensing devices.

Вплив комплексу біопротекторів на динаміку активності амінотрансфераз у сироватці крові за умов окремої та одночасної дії свинцю і фтору (експериментальні дослідження)

Lead and fluoride enter the human body individually and simultaneously through water, food, and air, disrupting metabolic processes in liver tissue. The sequential application of a bioprotective complex that reduce the negative effects of lead and fluoride has been insufficiently studied. The aim of the study is to investigate the changes over time in the activity of alanine aminotransferase and aspartate aminotransferase in the blood serum of laboratory animals under prolonged separate and simultaneous exposure to lead and fluoride, without application of bioprotective agents and during the sequential addition of pectin, pectin with calcium, and a complex of pectin, calcium, and antioxidants to the standard animal diet. Materials and Methods. Four clinical trial series were conducted on white rats, each following an orthogonal design 22. Aqueous solutions of Pb(NO3)2 and NaF were administered orally separately and simultaneously over 30 days: in the first series without bioprotective agents, in the second with pectin, in the third with pectin and calcium, and in the fourth with a complex of pectin, calcium, and antioxidants – C, E, and β-carotene vitamins, and selenium. The activity of alanine and aspartate aminotransferases was measured in blood serum on days 3, 15, and 30 in the first and second series of experiments, and on day 30 in the third and fourth series. Results. Separate and simultaneous oral administration of lead and fluoride to white rats leads to increased activity of alanine and aspartate aminotransferases in blood serum. The maximum changes were observed at the end of the experiment. The protective role of pectin alone under long-term fluoride expos ure and simultaneous exposure to lead and fluoride was not sufficiently effective. Concurrent intake of calcium and pectin significantly reduced the increase in transaminase activity, indicating an improvement in the metabolic and functional state of the liver. The application of the bioprotective complex restored the aminotransferase activity in blood serum. Conclusions. The complex of pectin with calcium and antioxidants can be considered an optimal means of correcting aminotransferase activity in blood serum under prolonged separate and simultaneous exposure to lead and fluoride.

The genetic resistance of sows to Escherichia coli F4 adhesion reduces their response to a vaccine containing F4 fimbriae but does not affect the preweaning performance of their susceptible piglets.

Pigs without intestinal receptors for F4 fimbriae are congenitally resistant to F4 fimbriae-bearing enterotoxigenic Escherichia coli (ETEC F4). In general, 50 % and 100 % of piglets born to resistant (RR) sows crossed with hetero- or homozygous susceptible (SR, SS) boars, respectively, are susceptible but do not receive colostral antibodies against F4 fimbriae unless the sows have been vaccinated. The question arises as to whether resistant sows produce protective amounts of F4 antifimbrial antibodies after vaccination. The serum and colostrum antibody titres of 12 resistant and 12 susceptible vaccinated gilts were compared. The effect of the receptor status of the dam and sire on the preweaning performance of 5027 piglets was evaluated using Agroscope's recordings. The sows of the experimental herd, where ETEC F4 was circulating, were vaccinated against ETEC twice during the first pregnancy and once during each following pregnancy. The log2 transformed F4 antibody titres in the serum obtained after the second vaccine injection as well as in the colostrum of the 12 resistant animals were lower than the titres of the susceptible animals (serum: F4ab 11,19 ± 1,44 vs. 12,18 ± 1,33, P = 0,096; F4ac 10,03 ± 1,58 vs. 11,59 ± 1,43, P = 0,019; colostrum: F4ab 12,20 ± 2,41 vs. 14,02 ± 1,31, P = 0,033; F4ac 10,93 ± 2,46 vs. 13,03 ± 5,21, P = 0,006). The heat labile enterotoxin (LT) antibody titres after vaccination did not differ between susceptible and resistant animals (p > 0,10). Preweaning mortality in the offspring of RR sows × SS boars was slightly lower than in the offspring of SS sows × RR boars (P = 0,04), suggesting that the disease risk of susceptible piglets born to vaccinated resistant sows was not increased, even though they received colostrum with a slightly reduced content of antibody against F4 fimbriae.