Eleutherococcus divaricatus Fruits Decrease Hyaluronidase Activity in Blood Serum and Protect from Antioxidative Damages in In Vitro Model.

Fruits are very important dietary components and a source of biologically active compounds used in nutritional pharmacology. Particularly due to the presence of polyphenolic compounds, fruits play an important role in the prevention of diseases of civilization. Therefore, it is important to study the phytochemicals and biological activity of fruits, especially those with a long-standing use in ethnomedicine. In this study, we determined the chemical profile and biological activity of a methanolic extract of the Eleutherococcus divaricatus fruits. Amongst nine polyphenols studied, only chlorogenic acid, protocatechuic acid, and eleutheroside E have been detected. The extract showed a weak anti-hyaluronidase activity from bovine testicular in a range of 9.06-37.70% and quite high for human serum hyaluronidase from children diagnosed with acute leukemia in a range of 76-86%. A weak anti-tyrosinase activity was obtained in a range of 2.94-12.46%. Moreover, the extract showed antioxidant properties against DPPH radical, ABTS radical, and O2•-. In addition, the antioxidant activity of the extract was evaluated by FRAP assay and Fe2+ ion chelation assay. These preliminary studies partially justify the traditional use of the plant in inflammatory- and immune-related diseases, in which hyaluronidase and free radicals can participate. A difference in human serum hyaluronidase inhibition may result from the inter-patient variability. Regardless of that, the results mean that polyphenolic compounds may stimulate activity of hyaluronidase, as well as to protect cells from the oxidative damages. However, further studies in ex vivo and in vivo models are needed, including blood isolated from a larger number of patients.

Insights into the Serum Metabolic Adaptations in Response to Inspiratory Muscle Training: A Metabolomic Approach Based on 1H NMR and UHPLC-HRMS/MS.

Inspiratory muscle training (IMT) is known to promote physiological benefits and improve physical performance in endurance sports activities. However, the metabolic adaptations promoted by different IMT prescribing strategies remain unclear. In this work, a longitudinal, randomized, double-blind, sham-controlled, parallel trial was performed to investigate the effects of 11 weeks (3 days·week-1) of IMT at different exercise intensities on the serum metabolomics profile and its main regulated metabolic pathways. Twenty-eight healthy male recreational cyclists (30.4 ± 6.5 years) were randomized into three groups: sham (6 cm·H2O of inspiratory pressure, n = 7), moderate-intensity (MI group, 60% maximal inspiratory pressure (MIP), n = 11) and high-intensity (HI group, 85-90% MIP, n = 10). Blood serum samples were collected before and after 11 weeks of IMT and analyzed by 1H NMR and UHPLC-HRMS/MS. Data were analyzed using linear mixed models and metabolite set enrichment analysis. The 1H NMR and UHPLC-HRMS/MS techniques resulted in 46 and 200 compounds, respectively. These results showed that ketone body metabolism, fatty acid biosynthesis, and aminoacyl-tRNA biosynthesis were upregulated after IMT, while alpha linolenic acid and linoleic acid metabolism as well as biosynthesis of unsaturated fatty acids were downregulated. The MI group presented higher MIP, Tryptophan, and Valine levels but decreased 2-Hydroxybutyrate levels when compared to the other two studied groups. These results suggest an increase in the oxidative metabolic processes after IMT at different intensities with additional evidence for the upregulation of essential amino acid metabolism in the MI group accompanied by greater improvement in respiratory muscle strength.

Estimating IgG concentration directly by radial immunodiffusion or indirectly by refractometry measure of serum total protein lack precision.

OBJECTIVE: To establish and compare the precision of serum total protein (STP) measured by an optical refractometer to the precision of IgG concentrations measured using radial immunodiffusion (RID), the reference test for quantifying IgG in neonatal calves. SAMPLE: 6 sera with previously measured IgG concentration using RID from neonatal beef calves were selected from 3 stratum: low-serum IgG stratum between >5.0 and <15.0g/L(n = 4); moderate-serum IgG stratum between 35.0-45.0g/L(n = 1); high-serum IgG stratum between 60.0-70.0g/L(n = 1). METHODS: STP was measured 13 times with an optical refractometer. IgG concentrations were measured 28 times with a commercial bovine IgG RID for each sera. The homogeneity of variance within the tests was evaluated with the Levene test (α = 0.10). Unrestricted random sampling bootstrapping (5,000 repetitions) was used to calculate the coefficient of variation (CV) for each serum and test. The homogeneity of variance between simulated test CVs by serum was evaluated (α = 0.10). Differences between simulated test CV by serum were assessed with the Kruskal-Wallis test (α = 0.05). RESULTS: No difference was observed in the variance for STP between sera (P = .39). The average CV for STP was 4.2%, 10.1% for the low IgG stratum, and 15.5% for the moderate/high IgG stratum. Variance differed in serum IgG concentration (P < .0001). Serum with higher IgG concentrations had more variance. Simulated CV for STP and IgG had homogeneity of variance for only 1 sera (P = .31). STP had a smaller CV compared to IgG for every serum (P < .0001). CLINICAL RELEVANCE: Estimating IgG concentration directly by RID or indirectly by STP lacks the precision that might affect diagnostic interpretation regarding a calf's absorption of maternal antibodies.

The Role of Selenium on the Status of Mineral Elements and Some Blood Parameters of Blood Serum of Lambs.

Selenium is one of the compounds belonging to the trace minerals group, which needs less than 100 mg/day. This element is one of the main constituents of selenoproteins, and the function of selenoproteins is to help make DNA and protect cells from damage and infection. This experiment aimed to evaluate the effect of different sources of selenium on some mineral elements in the blood serum of lambs. This experiment was conducted using twenty 4-month-old lambs with an average weight of 37±2.2 kg, 4 treatments, and 5 replications in a completely randomized design (CRD). The treatments tested included control, sodium selenite, nano selenium, and VitEsel. The experiment duration was 30 days, and blood sampling of lambs was performed at the beginning of the experiment (zero), 15, and 30 days. Different sources of selenium significantly affected the concentrations of iron, copper, and zinc (P<0.05). Different sources of selenium in this experiment decreased the concentration of iron and copper and increased the concentration of zinc and plasma selenium in different periods (P<0.05). Using different sources of selenium changed the concentration of the studied elements and showed the difference in their bioavailability.

Effect of Laminaria digitata dietary inclusion and CAZyme supplementation on blood cells, serum metabolites and hepatic lipids and minerals of weaned piglets.

Seaweeds, such as Laminaria digitata, are a sustainable alternative to conventional feedstuffs for weaned piglet diets, improving their health and mitigating environmental impacts. L. digitata has a complex cell wall that can be difficult for monogastrics to digest. However, carbohydrate-active enzymes (CAZymes) such as Rovabio® Excel AP and alginate lyase can help break down these polysaccharides and render intracellular nutrients more accessible. This study aimed to evaluate the impact of 10% L. digitata feed inclusion and CAZyme supplementation on piglet blood cells, serum metabolites, liver lipid and mineral profiles. Forty weaned piglets were randomly assigned to one of four diets (n = 10 each): a control diet, 10% L. digitata (LA), 10% L. digitata + 0.005% Rovabio® Excel AP (LAR), and 10% L. digitata + 0.01% alginate lyase (LAL). After two weeks of trial, animals were slaughtered and liver and blood serum samples taken for analysis. The results showed that the LA and LAL diets increased blood lymphocytes, IgG and IgM, and decreased serum lipids, improving both cellular and humoral immune response and cardiovascular health. Dietary CAZymes reversed the anti-inflammatory and hematopoietic effects. Additionally, cortisol levels were reduced with seaweed inclusion compared to the control diet (P < 0.001). In the liver, total n-3 PUFA and n-6/n-3 ratio were increased and decreased, respectively, due to eicosapentaenoic acid and α-linolenic acid accumulation (P < 0.001). However, total liver mineral content was incorporated to a lesser extent with the combined seaweed and enzyme diets (P < 0.001), potentially indicating a negative effect on mineral bioavailability. Overall, results suggest that a 10% L. digitata inclusion can effectively improve piglet health by reducing stress during weaning, without the need for dietary CAZymes.

Metal concentrations in cerebrospinal fluid, blood, serum, plasma, hair, and nails in amyotrophic lateral sclerosis: A systematic review and meta-analysis.

BACKGROUND: Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease with progressive muscle wasting, paralysis, and respiratory failure. Whereas approximately 10-15 % of ALS cases are familial, the etiology of the remaining, sporadic ALS cases remains largely unknown. Environmental exposures have been suggested as causative factors for decades, and previous studies have found elevated concentrations of metals in ALS patients. PURPOSE: This meta-analysis aims to assess metal concentrations in body fluids and tissues of ALS patients. METHODS: We searched the MEDLINE and EMBASE databases on December 7th, 2022 for cross-sectional, case-control, and cohort studies which measure metal concentrations in whole blood, blood plasma, blood serum, cerebrospinal fluid (CSF), urine, erythrocytes, nail, and hair samples of ALS patients. Meta-analysis was then performed when three or more articles existed for a comparison. FINDINGS: Twenty-nine studies measuring 23 metals were included and 13 meta-analyses were performed from 4234 screened entries. The meta-analysis results showed elevated concentrations of lead and selenium. Lead, measured in whole blood in 6 studies, was significantly elevated by 2.88 µg/L (95 % CI: 0.83-4.93, p = 0.006) and lead, measured in CSF in 4 studies, was significantly elevated by 0.21 µg/L (95 % CI: 0.01 - 0.41, p = 0.04) in ALS patients when compared to controls. Selenium, measured in serum/plasma in 4 studies, was significantly elevated by 4.26 µg/L (95% CI: 0.73 - 7.79, p = 0.02) when compared to controls.Analyses of other metal concentrations showed no statistically significant difference between the groups. CONCLUSION: Lead has been discussed as a possible causative agent in ALS since 1850. Lead has been found in the spinal cord of ALS patients, and occupational exposure to lead is more common in ALS patients than in controls. Selenium in the form of neurotoxic selenite has been shown to geochemically correlate to ALS occurrence in Italy. Although no causal relationship can be established from the results of this meta-analysis, the findings suggest an involvement of lead and selenium in the pathophysiology of ALS. After a thorough meta-analysis of published studies on metal concentrations in ALS it can only be concluded that lead and selenium are elevated in ALS.

Spherotech-EDTA combined serum treatment reduces background more effectively as compared to One Lambda Adsorb Out™ and LIFECODES Serum Cleaner in Luminex-based solid-phase immunoassays for HLA antibody detection.

Spherotech (SPT) microparticles capture non-specific binding materials present in test serum, and EDTA removes the so called" prozone effect". This study presents a novel approach of combined SPT-EDTA serum treatment prior to Luminex HLA antibody testing to remove high background, and prozone effect in a single step process, and compared the efficacy of SPT-EDTA serum pre-treatment with AdsorbOut (ADS) and Serum Cleaner (SC) to reduce background in solid phase immunoassays (SPI). A total of 21 serum samples with a history of elevated negative control (NC) values ≥500, and 20 samples with normal NC values were included to assess the potential adverse effects. A problem of high background was noted in 25% of our samples in SPI. We observed 80% effectiveness in reducing NC values <500 with SPT-EDTA serum pre-treatment compared to 72%, and 67% for ADS and SC-treated sera, respectively. Interestingly, we found a strong correlation in antibody-binding levels between SPT versus ADS; and ADS versus SC-treated sera for both phenotype and single antigen bead assays (p < 0.001). No adverse effect was noted on NC, positive control (PC) values, PC/NC ratios in the upfront use of SPT-EDTA as compared to EDTA alone. Our data revealed that combined SPT-EDTA treated sera is more effective than ADS, and SC in reducing high background in SPI. Taken together, SPT-EDTA serum treatment prior to Luminex HLA Ab testing is cost-effective, our laboratory saves nearly 30% of the annual total cost for Ab testing and improved test turnaround time by two business days.

Thermodynamics-based rules of thumb to evaluate the interaction of chelators and kinetically-labile metal ions in blood serum and plasma.

Metal ions play a very important role in nature and their homeostasis is crucial. A lot of metal-related chemical research activities are ongoing that concern metal-based drugs or tools, such as chelation therapy, metal- and metabolite sensors, metallo-drugs and prodrugs, PET and MRI imaging agents, etc. In most of these cases, the applied chelator/ligand (L) or metal-ligand complex (M-L) has at least to pass the blood plasma to reach the target. Hence it is exposed to several metal-binding proteins (mainly serum albumin and transferrin) and to all essential metal ions (zinc, copper, iron, etc.). This holds also for studies in cultured cells when fetal calf serum is used in the medium. There is a risk that the applied compound (L or M-L) in the serum is transformed into a different entity, due to trans-metallation and/or ligand exchange reactions. This depends on the thermodynamics and kinetics. For kinetically-labile complexes, the complex stability with all the ligands and all metal ions present in serum is decisive in evaluating the thermodynamic driving force towards a certain fate of the chelator or metal-ligand complex. To consider that, an integrative view is needed on the stability constants, by taking into account all the metal ions present and all the main proteins to which they are bound, as well as the non-occupied metal binding site in proteins. Only then, a realistic estimation of the complex stability, and hence its potential fate, can be done. This perspective aims to provide a simple approach to estimate the thermodynamic stability of labile metal-ligand complexes in a blood plasma/serum environment. It gives a guideline to obtain an estimation of the plasma and serum complex stability and metal selectivity starting from the chemical stability constants of metal-ligand complexes. Although of high importance, it does not focus on the more complex kinetic aspects of metal-transfer reactions. The perspective should help for a better design of such compounds, to perform test tube assays which are relevant to the conditions in the plasma/serum and to be aware of the importance of ternary complexes, kinetics and competition experiments.

Fatty acid correlations with HOMA-IR and HOMA-% ß are differentially dictated by their serum free and total pools and flaxseed oil supplementation.

Objective. The intent of the present study was to test two hypotheses. The primary hypothesis was that there would be differences between blood serum individual free fatty acids (SIFFA) and serum individual total fatty acids (SITFA) in terms of their different relationships (correlations) to each of homeostatic model assessment-individual insulin resistance (HOMA-IR) and homeostatic model assessment-individual insulin resistance-percentage ß-cell function (HOMA-% ß) remaining in human type 2 diabetic patients with pre-flaxseed oil (FXO) and pre-safflower oil (SFO) administration. The secondary hypothesis was that FXO (rich in alpha-linolenic acid, ALA) supplementation would alter these correlations differently in the SIFFA and STIFFA pools in comparison with the placebo SFO (poor in ALA). Methods. Patients were recruited via a newspaper advertisement and two physicians. All patients came to visit 1 and three months later to visit 2. At visit 2, the subjects were randomly assigned (double-blind) to flaxseed or safflower oil (placebo) treatment for three months until visit 3. Results. There were pre-intervention differences in the SIFFA and STIFA pool's relationships with each of HOMA-IR and HOMA-% ß. These relatioships remained either unchanged or became significant after intervention (treatment or placebo). There was a negative correlation found between HOMA-IR and serum free ALA (SFALA) mol % after FXO. Serum total ALA (STALA) mol % had no significant correlations with HOMA-IR and HOMA- % ß before and after flaxseed oil administration. Conclusions. The SIFFA and SITFA pools have different relationships with HOMA-IR and HOMA-% ß for each of pre- and post-intervention. It is concluded that the data support both the primary and the secondary hypotheses indicating that they are correct.

Continuous In Vitro Culture of Babesia duncani in a Serum-Free Medium.

Human babesiosis is an emerging tick-borne disease, caused by haemoprotozoa genus of Babesia. Cases of transfusion-transmitted and naturally acquired Babesia infection have been reported worldwide in recent years and causing a serious public health problem. Babesia duncani is one of the important pathogens of human babesiosis, which seriously endangers human health. The in vitro culture systems of B. duncani have been previously established, and it requires fetal bovine serum (FBS) to support long-term proliferation. However, there are no studies on serum-free in vitro culture of B. duncani. In this study, we reported that B. duncani achieved long-term serum-free culture in VP-SFM AGTTM (VP-SFM) supplemented with AlbuMaxTM I. The effect of adding different dilutions of AlbuMaxTM I to VP-SFM showed that 2 mg/mL AlbuMaxTM I had the best B. duncani growth curve with a maximum percentage of parasitized erythrocytes (PPE) of over 40%, and it can be used for long-term in vitro culture of B. duncani. However, the commonly used 20% serum-supplemented medium only achieves 20% PPE. Clearly, VP-SFM with 2 mg/mL AlbuMaxTM I (VP-SFMA) is more suitable for the in vitro proliferation of B. duncani. VP-SFM supplemented with CD lipid mixture was also tested, and the results showed it could support the parasite growth at 1:100 dilution with the highest PPE of 40%, which is similar to that of 2 mg/mL AlbuMaxTM I. However, the CD lipid mixture was only able to support the in vitro culture of B. duncani for 8 generations, while VP-SFMA could be used for long-term culture. To test the pathogenicity, the VP-SFMA cultured B. duncani was also subjected to hamster infection. Results showed that the hamster developed dyspnea and chills on day 7 with 30% PPE before treatment, which is similar to the symptoms with un-cultured B. duncani. This study develops a unique and reliable basis for further understanding of the physiological mechanisms, growth characteristics, and pathogenesis of babesiosis, and provides good laboratory material for the development of drugs or vaccines for human babesiosis and possibly other parasitic diseases.

The association between vitamin D serum levels, supplementation, and suicide attempts and intentional self-harm.

OBJECTIVES: The purpose of this study is to determine the associations between Vitamin D supplementation, 25(OH) blood serum levels, suicide attempts, and intentional self-harm in a population of veterans in the Department of Veterans Affairs (VA). METHODS: A retrospective cohort study of US Veterans supplemented with Vitamin D. Veterans with any Vitamin D3 (cholecalciferol) or Vitamin D2 (ergocalciferol) fill between 2010 and 2018 were matched 1:1 to untreated control veterans having similar demographics and medical histories. Cox proportional hazards regression was used to estimate the time from the first Vitamin D3 (cholecalciferol) or Vitamin D2 (ergocalciferol) prescription fill to the first suicide attempt or intentional self-harm. Analyses were repeated in stratified samples to measure associations by race (Black or White), gender (male or female), blood levels (0-19 ng/ml, 20-39 ng/ml, and 40+ ng/ml), and average daily dosage. RESULTS: Vitamin D3 and D2 supplementation were associated with a 45% and 48% lower risk of suicide attempt and self-harm ((D2 Hazard Ratio (HR) = 0.512, [95% CI, 0.457, 0.574]; D3 HR = 0.552, [95% CI, 0.511, 0.597])). Supplemented black veterans and veterans with 0-19 ng/ml vitamin D serum levels were at ~64% lower risk relative to controls (Black Veteran HR: 0.362 [95% CI: 0.298,0.440]; 0-19 ng/ml HR: 0.359 [95% CI: 0.215,0.598]). Supplementation with higher vitamin D dosages was associated with greater risk reductions than lower dosages (Log Average Dosage HR: 0.837 [95% CI: 0.779,0.900]). CONCLUSIONS: Vitamin D supplementation was associated with a reduced risk of suicide attempt and self-harm in Veterans, especially in veterans with low blood serum levels and Black veterans.

Serum 25-hydroxyvitamin D and fatty acids in relation to the risk of microbial infections in children: The TRIGR Divia study.

BACKGROUND & AIMS: Nutrient status may affect the risk of microbial infections and play a role in modulating the immune response against such infections. The aim of this study was to determine whether serum 25-hydroxyvitamin D [25(OH)D] and serum fatty acids in infancy are associated with microbial infections by the age of 18 months. METHODS: Altogether 576 newborn infants from Trial to Reduce IDDM in the Genetically at Risk (TRIGR) born between 2002 and 2007 were included. The concentration of 25(OH)D vitamin and proportions of 26 fatty acids (presented as % of total fatty acids) were analyzed in cord blood serum and in sera taken at 6, 12, and 18 months of age. The cord blood samples and mean of 6-18-month values were used as exposures. Infections were detected by screening IgG antibodies against 10 microbes using enzyme immunoassay and antibodies against 6 coxsackievirus B serotypes by plaque neutralization assay in serum samples taken at 18 months of age. RESULTS: A higher proportion of n-3 polyunsaturated fatty acids (PUFAs) and especially long-chain n-3 PUFAs at birth and at the age of 6-18 months was associated with decreased risk of coxsackievirus B2 infection unadjusted and adjusted for region, case-control status, and maternal type 1 diabetes. Higher proportion of docosapentaenoic acid (DPA, 22:5 n-3) at birth was associated with a decreased risk of respiratory syncytial virus infection. 25(OH)D vitamin concentration was not consistently associated with the risk of infections. When only infected children were included docosahexaenoic acid (DHA, 22:6 n-3) and arachidonic acid (20:4 n-6) proportions were positively associated with IgG antibody levels against influenza A virus. 25(OH)D vitamin concentration showed an inverse association with rotavirus IgG levels among children with rotavirus seropositivity. CONCLUSIONS: In young children with increased susceptibility to type 1 diabetes, long-chain n-3 PUFAs may influence the risk of viral infections and immune response against the infections. However, this association may depend on the type of virus suggesting virus-specific effects.

A Pilot Study on the Association of Lead, 8-Hydroxyguanine, and Malondialdehyde Levels in Opium Addicts' Blood Serum with Illicit Drug Use and Non-Addict Persons.

While a large body of literature has shown the health problems of illicit drug use, research is needed on how substance abuse impacts DNA damage and contaminants in blood, especially given Pb-contaminated opium. This pilot study aimed to evaluate the levels of lead (Pb), 8-hydroxy di-guanine (8-oxo-Gua), and malondialdehyde (MDA) in the blood serum of opium addicts and non-addict people. The current study is a case-control study with a cross-sectional design. A sample of 50 opium-addicted and non-addict adults were chosen for this study using convenience and random sampling methods. Participants were divided into two groups: addicts and non-addicts. The atomic absorption spectroscopy method was used to measure the quantity of Pb, and the Enzyme-Linked Immunosorbent Assay (ELISA) method was used to measure the amount of 8-oxo-Gua and MDA. The data were analyzed using an independent t-test. The results show that the amount of Pb in the blood serum of addicted women and men was higher than levels in non-addict men and women, for the study participants (p-value = 0.001). Blood levels were not significantly different between addicts and non-addicts for men or women for 8-oxo-Gua (p-value = 0.647 for women and p-value = 0.785 for men) and MDA (p-value = 0.867 for women and p-value = 0.995 for men). In general, addicts' blood Pb levels were found to be substantially higher than those of normal non-addict persons in this pilot study. As a result, testing for blood Pb levels in addicts may be informative in instances when symptoms are inconclusive.

[Q-markers of Yuquan Capsules based on serum pharmacochemistry of Chinese medicine].

This study analyzed the quality markers(Q-markers) of Yuquan Capsules(YQC) based on serum pharmacochemistry of Chinese medicine and detected the components and metabolites of YQC absorbed into the blood by UPLC-Q-TOF-MS and UNIFI systems. As a result, 32 components of YQC were detected, including 17 prototype components and 15 metabolized components. Among them, 12 prototype components(ginsenoside Rh_2, genistein, formononetin, puerarin, daidzein, schizandrin A, schizandrin B, schizandrin C, schizandrol A, schizandrol B, gomisin D, and ononin) and 12 metabolized components(ginsenoside Rg_1, ginsenoside Rg_2, ginsenoside Rg_3, ginsenoside Ro, 3'-methoxypuerarin, daidzin, astragaloside Ⅱ, astragaloside Ⅳ, glycyrrhizic acid, liquiritigenin, isoliquiritin, and verbascoside) showed inhibitory effects and pharmacological activities against diabetes, and these 24 blood-entering components against diabetes were identified as Q-markers of YQC.

Extracts of Periplaneta americana alleviate hepatic fibrosis by affecting hepatic TGF-ß and NF-κB expression in rats with pig serum-induced liver fibrosis.

INTRODUCTION: Liver fibrosis is caused by continuous wound healing responses to various harmful stimuli, including viral infection, drugs, alcohol, and autoimmune liver disease. The purpose of this study was to examine the effects of extracts of Periplaneta americana (EPA) in rats with pig serum-induced liver fibrosis to preliminarily assess the antifibrotic effect of EPA. MATERIAL AND METHODS: Seventy rats were randomly divided into 7 groups (10 rats in each group): HC, the healthy control group; FC, the fibrotic control group; TL, low-dose EPA treatment group group; TM, medium-dose EPA group; TH, high-dose EPA treatment group; TC1, Panax notoginseng/Salvia mitiorrhiza treatment control group 1; TC2, colchicine treatment control group 2. TC1 and TC2 were used as the positive control to demonstrate the difference between EPA and the effects of other compounds. The liver fibrosis model was induced by intraperitoneal injection of 0.5 mL pig serum twice a week for 13 weeks in all groups except for the HC group. The hepatic fibrosis model was established at the 7th week, and followingly, the corresponding compounds were administered once a day in all groups for 6 weeks. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity was determined in rat blood serum. We also measured liver fibrosis-related serum markers, including hyaluronic acid (HA), mucin layer (LN), type III pre-collagen (PC-III) and type IV collagen (IV-C). Hematoxylin and eosin (H&E) and Masson stainings were used to assess liver morphology and determine the stage of fibrosis. Immunohistochemistry was used to detect the protein expression of NF-κB, α-smooth muscle actin (α-SMA), transforming growth factor-ß1 (TGF-ß1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in rat liver tissue. RESULTS: Compared with that of the HC group, the liver tissue of the FC group presented obvious liver damage and collagen deposition. The serum levels of ALT, AST, HA, LN, PC-III and IV-C and the expression of NF-κB, α-SMA, TGF-ß1 and TIMP-1 in the FC group were significantly higher than those in the HC group, the EPA treatment groups, the TC1 group and the TC2 group (P < 0.01). The levels of serum ALT, AST, HA, LN, PC-III and IV-C and the expression of α-SMA, NF-κB, TGF-ß1 and TIMP-1 in the TL, TC1 and TC2 groups were significantly higher than those TM and TH groups (P < 0.05). EPA treatment significantly improved liver function, decreased collagen deposition and reversed the pathological changes related to liver fibrosis. CONCLUSIONS: We found that EPA could reduce liver inflammation, suppress liver cell degeneration and necrosis, and reduce the formation of liver fibrous tissue. Its mechanism might be associated with inhibiting the expression of TGF-ß1, TIMP-1, NF-κB and α-SMA to block signal transduction pathways in the hepatic fibrosis process. Therefore, EPA, as a traditional Chinese medicine, might be potentially used to prevent and treat hepatic fibrosis in the future. However, further more experiments are necessary to verify its effectiveness and possible signaling pathways.

Verificación del desempeño general del método Elecsys® Chagas en la detección de anticuerpos específicos contra Trypanosoma cruzi / Verification of the general performance of the Elecsys® Chagas method in the detection of specific antibodies against Trypanosoma cruzi

Resumen La enfermedad de Chagas es una parasitosis producida por Trypanosoma cruzi, prevalente principalmente en el continente americano, y observada en regiones no endémicas, producto de viajes y migraciones. El objetivo de este estudio fue comparar el desempeño del ensayo Elecsys® Chagas (Roche Diagnostics Alemania) (ECLIA) para el diagnóstico de la infección chagásica crónica con el método estándar y evaluar su posible empleo en reemplazo del método automatizado existente. Se estudiaron 77 muestras de sueros pertenecientes a pacientes con diagnóstico presuntivo de enfermedad de Chagas, procesadas por los distintos métodos disponibles en la Sección Parasitología del Hospital Muñiz: inmunoensayo quimioluminiscente de micropartículas (CMIA) (Abbott), enzimoinmunoanálisis de adsorción (ELISA) (Wiener) y hemaglutinación indirecta (HAI) (Lab. Lemos S.R.L.). Los resultados de los métodos ELISA y HAI fueron comparados con los obtenidos en la prueba ECLIA, y estos a su vez con el método automatizado disponible. De las muestras analizadas, 22 (28,57%) presentaron IgG anti-T. cruzi y 55 (71,43%) resultaron negativas. Con el método ECLIA se logró un 100% en los parámetros de desempeño, con diferencias en los intervalos de confianza. La razón de verosimilitud positiva y la razón de verosimilitud negativa clasificaron al ensayo como excelente y la potencia global del test apoyó esa afirmación. Los métodos inmunológicos automatizados ayudan a la performance diagnóstica en la etapa crónica de la enfermedad de Chagas, permiten minimizar errores, favorecen la velocidad de emisión de los resultados y, debido a su alta sensibilidad y especificidad, en ciertos escenarios podrían proponerse para usar como única técnica.

Abstract Chagas disease is a parasitosis caused by Trypanosoma cruzi, prevalent mainly in the American continent, and observed in non-endemic regions as a result of travel and migration. The objective of this study was to compare the performance of the Elecsys® Chagas (Roche Diagnostics Alemania) (ECLIA) assay for the diagnosis of chronic Chagas infection with the diagnostic standard, and to evaluate its possible use as a replacement for the existing automated method. A total of 77 serum samples belonging to patients with a presumptive diagnosis of Chagas disease were evaluated, processed by the different methods available in the Parasitology Section of Hospital Muñiz: microparticle chemiluminescent immunoassay (CMIA) (Abbott), enzyme-linked immunosorbent assay (ELISA) (Wiener) and indirect hemagglutination (HAI) (Lab. Lemos S.R.L). The results of the ELISA and HAI methods were compared with those obtained in the ECLIA test, and these in turn with the available automated method. Of the samples analysed, 22 (28.57%) presented IgG anti-T. cruzi and 55 (71.43%) were negative. With the ECLIA method, 100% was achieved in the performance parameters, with differences in the confidence intervals. The positive likelihood ratio and the negative likelihood ratio classify the essay as excellent, and the overall power of the test supports this statement. Automated immunological methods help diagnostic performance in the chronic stage of Chagas disease, allow minimising errors, favour the speed of issuance of results, and due to the high sensitivity and specificity, in certain scenarios, they could be proposed for use as single technique.

Resumo A doença de Chagas é uma parasitose causada pelo Trypanosoma cruzi, prevalente principalmente no continente americano, e observada em regiões não endêmicas em decorrência de viagens e migrações. O objetivo deste estudo foi comparar o desempenho do ensaio Elecsys® Chagas (Roche Diagnostics Alemanha) (ECLIA) para o diagnóstico da infecção crônica de Chagas com o método padrão e avaliar seu possível uso em substituição do método automatizado existente. Foram avaliadas 77 amostras de soro pertencentes a pacientes com diagnóstico presuntivo de doença de Chagas, processadas pelos diferentes métodos disponíveis na Seção de Parasitologia do Hospital Muñiz: imunoensaio quimioluminescente de micropartículas (CMIA) (Abbott), ensaio imunoenzimático de adsorção (ELISA) (Wiener) e hemaglutinação indireta (HAI) (Lab. Lemos S.R.L). Os resultados dos métodos ELISA e HAI foram comparados com os obtidos no teste ECLIA, e estes por sua vez com o método automatizado disponível. Das amostras analisadas, 22 (28,57%) apresentaram IgG anti-T. cruzi e 55 (71,43%) foram negativos. Com o método ECLIA, foram obtidos 100% nos parâmetros de desempenho, com diferenças nos intervalos de confiança. A razão de verossimilhança positiva e a razão de verossimilhança negativa classificam o ensaio como excelente, e a potencia geral do teste conformou essa afirmação. Os métodos imunológicos automatizados auxiliam no desempenho diagnóstico na fase crônica da doença de Chagas, permitem minimizar erros, favorecem a rapidez na emissão dos resultados e, devido à alta sensibilidade e especificidade, em determinados cenários, poderiam ser propostos para uso como técnica única.

[Comparative study of rat serum pharmacochemistry between Puerariae Lobatae Radix and Puerariae Thomsonii Radix based on UPLC-Q-TOF-MS].

UPLC-Q-TOF-MS and serum pharmacochemistry were employed to study the migrating components in rat sera after intragastric administration of the water extracts of Puerariae Lobatae Radix(PLR) and Puerariae Thomsonii Radix(PTR). After the respective intragastric administration of PLR and PTR extracts, blood samples were collected from the orbital vein. The serum samples were treated by protein precipitation method with methanol and acetonitrile at a ratio of 1∶1 and then passed through Agilent ZORBAX RRHD SB-C_(18) column(3 mm×100 mm, 1.8 µm) and Agilent SB-C_(18) pre-column(3 mm×5 mm, 1.8 µm) with 0.1% formic acid aqueous solution(A)-acetonitrile(B) as the mobile phase. The elution was performed at the flow rate of 0.25 mL·min~(-1), the column temperature of 40 ℃, and the injection volume of 2 µL. By comparison of the total ion chromatogram and secondary fragment ion information of PLR and PTR water extracts, PLR-and PTR-containing sera, and blank serum, we found 42 migrating components(including 17 prototype components and 25 metabolites) in the sera of rats treated with PLR and 35 migrating components(including 15 prototype components and 20 metabolites) in the sera of rats treated with PTR. Thirty-three common components were shared by the two treatments, including 13 prototype components and 20 metabolites. The differences of migrating components in the PLR-and PTR-treated rat sera provide a scientific basis for further study of the active components and quality markers of PLR and PTR.

[The study of blood serum trace elements in comparison with the structural and functional characteristics of goiter and the carriage of antithyroid antibodies in some regions of Russia].

The study of the sufficiency with essential trace elements in the context of thyroid pathology is especially relevant, since the development and progression of thyroid diseases have various pathogenetic mechanisms, which largest proportion is provoked by insufficient intake of key trace elements, primarily iodine. The aim of the research was obtaining primary data on the sufficiency with essential trace elements of the population in the some regions of the Russian Federation, comparing the data obtained with the prevalence of goiter and the carriage of antibodies to thyroid peroxidase (AT-TPO). Material and methods. The Cross-Sectional Population Study was conducted in medical institutions of the Republics of Crimea, Republics of Tuva and Bryansk region (persons applied for outpatient consultations). The scope of the study was 387 people aged 18 to 65 years (the average age was 40±5 years). All of them underwent: collection of medical history, endocrinologist examination with palpation of the thyroid gland (TG), ultrasound examination of TG, blood sampling to assess the levels of thyroid-stimulating hormone (TSH), AT-TPO, iodine, selenium, zinc. Ultrasound of the TG was performed in the supine position using a portable ultrasound machine LOGIQe (China) with a multi-frequency linear probe 10-15 MHz. During the study, the volume of the thyroid gland, the presence of nodular formations and their characteristics according to the TIRADS classification, TG structure and its echogenicity were assessed. Iodine, selenium and zinc level were measured using tandem mass spectrometry with ionization in inductively coupled plasma (Agilent 8900 ICP-MS Triple Quad). TSH and Ab-TPO were determined by chemiluminescent immunoassay on the Architect i2000 automatic analyzer. Results. All regions are comparable in terms of iodine availability, but differ in the severity of natural iodine deficiency. The median concentration of iodine in blood serum practically coincided in all three regions, amounting to 39.8 µg/l in the Republic of Crimea; 38.8 µg/l in the Republic of Tyva and 43.4 µg/l in the Bryansk region. Comparing these results with foreign data, as well as with our results on urinary iodine level, obtained by routine cerium-arsenite method, we can assume that iodine content in blood serum corresponded to the lower limit of the normal target values. The median concentration of selenium in blood serum also practically coincided in all three regions, amounting to 68.2, 72.1 and 62.8 µg/l, respectively, and probably meets the criteria for optimality. The median serum concentration of zinc in Bryansk region residents corresponded to a deficiency (491.3 µg/l) and was significantly lower compared to the indicator in the Republics of Crimea and Tyva (1633.2 and 1667.6 µg/l, respectively, values above optimal). Among the examined, the proportion of people with AB-TPO carriership was 23.5% (Republic of Crimea - 20.9%, Republic of Tuva - 26.4%, Bryansk region - 20.7%), with impaired thyroid function - 9.6% (6.2% - hypothyroidism, 3.4% - thyrotoxicosis). The frequency of goiter varied from 10 to 13.8%. There were no significant differences in the frequency of occurrence, ultrasonic characteristics of goiter and thyroid nodules by regions. Conclusion. However, the results obtained do not allow to unequivocally judge the absence of micronutrient deficiency in the population of the regions of the Russian Federation, since used the ICP-MS method requires the development of our own reference values of trace elements; in the absence of the latter, data from similar foreign studies were taken as standards. In addition, in further studies it will be important to take into account additional criteria for assessing trace element deficiency, for example, activity of superoxide dismutase and alkaline phosphatase as indirect markers of zinc deficiency, etc. Correlation between blood serum concentration of trace elements and the frequency and structural characteristics of goiter, the carriage of Ab-TPO, and dysfunction of the TG was not revealed.

Effect of different levels of pomegranate peel powder and probiotic supplementation on growth, carcass traits, blood serum metabolites, antioxidant status and meat quality of broilers.

The present study aimed to investigate the effect of dietary pomegranate peel powder (PPP) and probiotic bacteria (PB) on the growth rate, carcass traits, blood serum metabolites, and meat quality of Ross broiler chickens during 1-5 weeks of age. A total of 480 unsexed Ross broiler chicks 7-days old with the beginning bodyweight of 110.58 ± 0.17 g were employed in a complete randomized purpose trail with eight groups; 60 chicks in six replicates (8 × 6 × 10). The dietary treatments were as follows: NC: negative group (without additives) group one; PC: positive control (main diet + 0.5 g Colostin antibiotic/kg feed); PPP 3-5: basal diet + 2, 3, 4 g pomegranate peel powder/kg diet and PPP 6-8: basal diet + 2, 3, 4 g pomegranate peel powder + 1 cm3 probiotic (Bacillus toyonensis (BT)/kg diet, respectively. The results showed that live body weight (LBW) at five weeks and body weight gain (BWG) during 1-5 weeks of age were affected by adding PPP in the ration and the good grads of PPP were 2 and 4 g PPP without PB/kg diet compared to NC and PC, respectively. Otherwise, daily feed conception (DFC) and feed conversion ratio (FCR) were not affected by adding the different grads of PPP with or without PB, except the first period of DFC (1-3 weeks of old) were affected. Results showed a significant effect on all carcass characteristics studied, except gizzard and abdominal fat ratio were not influenced by the treatment used. Likewise, the addition of PPP to broiler chicken diets has a good effect on almost the blood serum metabolites, immunological parameters and quality of meat studied. In the end, the outcome of this study concluded that the addition of PPP to broiler diets has a good effect on the growth rate, blood serum metabolites, immunological parameters and the quality of meat as well as the health aspects.

From urban dust and marine sediment to Ginkgo biloba and human serum-a top ten list of Standard Reference Materials (SRMs).

During the past 40 years, the National Institute of Standards and Technology (NIST) has developed over 180 natural matrix Standard Reference Materials® (SRMs) for the determination of trace organic constituents in environmental, clinical, food, and dietary supplement matrices. A list of the Top Ten SRMs intended for organic analysis was identified based on selection criteria including analytical challenge to assign certified values, challenges in material preparation, novel matrices, longevity, widespread use, and unique design concept or intended use. The environmental matrix SRMs include air particulate matter, marine sediment, mussel tissue, and human serum with the focus on contaminants such as polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), chlorinated pesticides, and polybrominated diphenyl ethers (PBDEs). Human serum and plasma SRMs for clinical diagnostic markers including vitamin D metabolites represent clinical analysis, whereas infant formula, multivitamin/multielement tablets, and Ginkgo biloba constitute the food and dietary supplement matrices on the list. Each of the SRMs on the Top Ten list is discussed relative to the selection criteria and significance of the material, and several overall lessons learned are summarized.