Determination Of Serum Proteins Research Articles

Proteomic biomarker evaluation using antibody microarrays: association between analytical methods such as microarray and ELISA.

To evaluate the associations between analytical methods, such as microarray and enzyme-linked immunosorbent assay (ELISA); expedient cutoffs; and the lowest possible number of microarrays in analysis for target biomarker estimation in case-control studies. This study included 321 serum specimens, gathered in different case-control studies to test for atherosclerosis and atrial fibrillation. Among them, 48 serum specimens were analyzed using microarray technology. We used ELISA and commercial kits for confirmation of the results. Three proteins-cadherin-P, neuronal nitric oxide synthase, and adenovirus fiber-were shown to have distinctly different values in the case group vs the control group. As a result, we used those proteins as the target for confirmation using our alternative analytical method. Also, these protein values represented the limiting range between the highest and lowest differences in case-control groups. The results of microarray assay were confirmed using ELISA and commercial kits in the same specimens, in which microarray profiling was performed, and also in separate large case-control groups. A 1.5-fold difference in the protein content, as measured using microarray technology, was shown to be sufficient for further investigation of the candidate proteins. As few as 3 microarrays were considered sufficient for perspective evaluation of the target proteins. Microarray serum profiling, therefore, provides semiquantitative determination of protein in serum.

Smartphone digital image colorimetry for quantification of serum proteins.

Colorimetric determination of proteins in serum is proposed based on the biuret method and replacing ultraviolet-visible spectrometric (UV-Vis) detection with a simple and affordable smartphone digital image colorimetric (SDIC) method. Optimum SDIC conditions were found as a detection wavelength of 555 nm, a region of interest of 1600 px2, and 9.0 cm between the detection camera and sample solution. Under the optimum conditions, the coefficient of determination was 0.9982 within a linear dynamic range of 0.022 to 0.35 g dL-1. The precision of the method based on the percent relative standard deviation was below 5%. The limit of detection and limit of quantitation were found to be 0.007 and 0.022 g dL-1 respectively, which were sufficient for the quantification of the total protein, albumin, and globulin in serum. The method was validated with an independent experiment using a UV-Vis method and both methods showed good statistical agreement, indicating the accuracy of the proposed SDIC method.

M251 Potential reference measurement procedure— Determination of c-reactive protein in serum by isotope dilution mass spectrometry based on enzyme digestion

M251 Potential reference measurement procedure— Determination of c-reactive protein in serum by isotope dilution mass spectrometry based on enzyme digestion

Determination of Interferon Gamma Protein in Serum of Breast Cancer Patients Using the ELISA

Interferon gamma (IFN-γ) is a cytokine involved in the induction and modulation of various immune responses. INF-γ has recently been associated with breast cancer and the stage of the disease. Also, it can be used as a cancer marker in determining disease activity. The ELISA test is a technique for identifying antigens in biological materials. The ELISA test is used for the detection of antibodies against target antigen through highly antibody-antigen interactions. In this study, the blood samples were taken from 88 Iraqis, 30 healthy controls, and 58 breast cancer patients (ranging from 35 to 52 years of age). During a visit to the AL Amal National Hospital for Cancer Treatment in Baghdad, Iraq, these patient samples were taken October 2020 - January 2021. Depending on the color changes to yellow, the results of this study were determined under wavelength of 450 nm to reveal the optical density (OD) via spectrophotometrically. The OD value is proportional to the amount of human IFN-γ in the sample of the healthy control and the cancer patients. The blood concentration of IFN-γ was substantially greater in women with breast cancer than in the control group (43.15 ± 4.68 pg/ml vs. 18.8 ± 3.89 pg/ml), respectively. Our results showed an elevation of Interferon-gamma (IFN-γ) in most of the serum samples from women with breast cancer. The conclusions of this study are the main findings, INF-y protein was employed as a marker and assistance in diagnosing breast cancer based on the results of the calculation of protein concentration from the ELISA test.

The content of the total protein, protein fractions and blood serum proteins in patients with different forms of lichen ruber planus

Purpose of the study. Establishing the role of processes of proteolysis of mixed saliva in the development and course of lichen planus of the oral mucosa.
 Materials and methods. A comprehensive examination of 102 patients with lichen planus aged 21 to 70 years and 20 people in the control group, whose age and sex composition corresponded to that in the study group. BioRad (USA) reagent kits were used to determine the total protein content of mixed saliva. The content of protein fractions of mixed saliva was determined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Determination of serum proteins in mixed saliva was performed by quantitative (cross) immunoelectrophoresis.
 Results. In patients with lichen planus, the overall proteolytic activity of mixed saliva increases with a significant increase in the concentration of α1proteinase inhibitor, especially in exudative hyperemic and erosiveulcerative forms of the disease. Diffusion of α1proteinase inhibitor into mixed saliva increases its antiproteolytic potential and has a protective character. The content of albumin and ceruloplasmin in the mixed saliva of patients with lichen planus increases depending on the severity of the disease: typical, hyperkeratotic, exudativehyperemic, erosiveulcerative.
 Conclusions. Mixed saliva of patients with lichen planus in contrast to patients in the control group is characterized by the predominance of low molecular weight proteins (20–79 kDa) over high molecular weight. The level of albumin, α1proteinase inhibitor and ceruloplasmin in the mixed saliva of patients with lichen planus increases and correlates with the severity of the disease. The content of IgA in the mixed saliva of patients with lichen planus increases, depending on the form of the disease.

Temporal changes in blood oxidative stress biomarkers across the menstrual cycle and with oral contraceptive use in active women.

To examine the temporal changes in blood oxidative stress biomarkers in recreationally-trained women that were naturally-cycling (WomenNC) or using oral contraceptives (WomenOC) across one month. Blood samples were acquired at three timepoints of the menstrual cycle (1: early-follicular, 2: late-follicular and 3: mid-luteal) and oral contraceptive packet (1: InactiveOC, 2: Mid-activeOC and 3: Late-activeOC) for determination of estradiol, progesterone, oxidative stress, C-reactive protein (CRP) and other cardiometabolic biomarkers in plasma and serum. There was a Group by Time effect on estradiol (p < 0.001, partial η2 = 0.64) and progesterone (p < 0.001, partial η2 = 0.77). Malondialdehyde, lipid hydroperoxides and CRP concentrations were higher in WomenOC during Late-activeOC compared to InactiveOC (+ 96%, + 23% and + 104%, respectively, p < 0.05). However, there were no changes in these biomarkers across the menstrual cycle in WomenNC (p > 0.05). At all timepoints (i.e., 1, 2 and 3), WomenOC had elevated lipid hydroperoxides (+ 28, + 48% and + 50%) and CRP (+ 71%, + 117% and + 130%) compared to WomenNC (p < 0.05, partial η2 > 0.25). There was no Group by Time effect on non-enzymatic antioxidants or glutathione peroxidase; however, glutathione peroxidase was lower in WomenOC, i.e., main effect of group (p < 0.05, partial η2 > 0.20). These findings demonstrate that WomenOC not only have higher oxidative stress and CRP than WomenNC, but also a transient increase across one month of habitual oral contraceptive use. Since changes in oxidative stress and CRP often relate to training stress and recovery, these outcomes may have implications to workload monitoring practices in female athletes.

Intranasal oxytocin affects the stress response to social isolation in sheep

Oxytocin (OT) is a neuropeptide hormone that modulates several social behaviors and can affect the anxiety and stress response. The aim of this study was to determine if administration of intranasal OT affects the stress response to social isolation in sheep. Twenty adult Merino ewes were assigned to two groups; 1) Control group (CG, n = 10), which received an intranasal administration of isotonic saline and 2) Oxytocin-treated group (OTG, n = 10), which received an intranasal administration of OT (24 IU) 40 min before the animals were placed in the social isolation test. During the social isolation test (10 min), the behavior of the sheep was recorded, and blood samples were obtained before and after the test for the determination of cortisol, glucose and serum proteins, and heart rate and surface temperature were recorded. The OTG ewes had a higher cortisol concentration (P = 0.04) after social isolation, tended to vocalize more (P = 0.06) during isolation, and tended to have lower globulin concentrations (P = 0.10) than the CG ewes. Contrary to what we expected, the administration of intranasal OT increased the stress response to social isolation in ewes, which was evidenced by endocrine (greater increase in cortisol concentration), physiological (a tendency to present lower concentration of globulins in blood) and behavioral (a tendency to vocalize more) indicators. This study suggests that the administration of intranasal OT increased the stress response to isolation possibly by strengthening the social bond among ewes.

A comparison of novel inactivation methods for production of a vaccine against Streptococcus agalactiae in Nile tilapia Oreochromis niloticus

Disease prevention and control are crucial for the development of intensive commercial aquaculture. For some fish species, several vaccines are available to contain the effects of pathogens. However, in Nile tilapia there are just a few vaccines available. The current study compared the efficacy of three inactivated vaccines against Streptococcus agalactiae; in Nile tilapia. Formaldehyde, hydrogen peroxide and pH manipulation were used to inactivate the bacteria. Nile tilapia fingerlings were distributed into six experimental groups in duplicate: T1: naïve group; T2: sham group (PBS-injected, non-challenged); T3: PBS-injected control group (PBS-injected, challenge-infected); T4: vaccinated with hydrogen peroxide-inactivated vaccine (vaccinated, challenge-infected); T5: vaccinated with formaldehyde-inactivated vaccine (vaccinated, challenge-infected); T6: vaccinated by pH manipulation (vaccinated, challenge-infected). Twenty-eight days after vaccination, groups T3, T4, T5 and T6 were challenged intraperitoneally with 0.1 mL of the bacteria and monitored for 21 days. Additionally, replicates of non-challenged fish were used for determination of total serum protein and anti-Streptococcus agalactiae IgM antibodies. The percentage of survival of the groups T1, T2, T3, T4, T5 and T6 were 100%, 100%, 62.5%, 80%, 75% and 85%, respectively. The group vaccinated by pH manipulation presented slight higher survival when compared to the control group (P > .05). However, there were no differences in survival between vaccinated groups. The relative percentage survival (RPS) in groups T4, T5 and T6 were 46.6%, 33.3% and 60%, respectively. No differences were observed for total serum proteins and anti-Streptococcus. agalactiae IgM antibodies. The results observed in this study indicate equivalent moderate vaccine efficacy between alternative inactivated (hydrogen peroxide and pH), and conventional formaldehyde vaccines, with slight higher values for the pH manipulated vaccine, which suggests that future studies should be performed for each alternative method in order to improve their performance.

Signal multi-amplified electrochemical biosensor for voltammetric determination of tau-441 protein in biological samples using carbon nanomaterials and gold nanoparticles to hint dementia.

A signal multi-amplified electrochemical biosensor was fabricated for tau-441 protein, a dementia biomarker. It utilizes a carbon nanocomposite film modified gold electrode. The carbon nanocomposite film was composed of multi-walled carbon nanotubes (MWCNTs), reduced graphene oxide (rGO), and chitosan (CS). For the nanocomposite film, rGO improved the dispersibility of MWCNTs, and the effective surface area of MWCNTs was increased. On the other hand, MWCNTs also increased the interlayer spacing of rGO, resulting in a thinner rGO layer. MWCNTs-rGO had a better conductivity than that of MWCNTs and rGO due tothe synergy effect. Biocompatible CS was employed for immobilization of the specific antibody. Tau-441 protein was modified with gold nanoparticles (AuNPs) for signal amplification again. The response of the electrochemical biosensor is linear in the range0.5-80fM (0.5, 1.5, 5, 10, 40, 80fM) with a limit of detection (LOD) of 0.46 fM, using differential pulse voltammetry (DPV) in a potential range of - 100-500mV. The biosensor was successfully applied to the analysis ofserum samples of 14 normal people, 14 mild cognitive impairment (MCI) patients, and 14 dementia patients. Graphical abstract Schematic representation of signal multi-amplified electrochemical biosensor for determination of tau-441 protein in human serum.

Application of Monoclonal Antibodies and Phage Display Technology for YB-1 Protein Analysis

Monoclonal antibodies against the recombinant YB-1 protein were obtained and characterized. These antibodies are capable of specific detection of YB-1 protein in the enzyme-linked immunosorbent assay, Western blot analysis, and immunocytochemistry. Using phage display technology, a peptide interacting with the YB-1 protein was found. Based on monoclonal antibodies МАYB-1-2 and a peptide as part of the bacteriophage structural protein, a test system was developed for the quantitative determination of YB-1 protein in blood serum. The detection limit of the developed test system is 0.3 ng/mL.

Acute bronchitis: answers to key questions.

The publication is devoted to the etiology and pharmacotherapy of acute bronchitis in adults. The authors obtained data on the etiology of the disease, indicating that in the clear majority of cases acute bronchitis is caused by various respiratory viruses (adenoviruses, influenza viruses and parainfluenza, and combinations of different viruses). The results of the study show that the starting prescription of antibacterial drugs for acute bronchitis is impractical, and the decision on their use should be based on objective data indicating the development of the bacterial process. A good tool for «distinguishing» a viral and bacterial infection is the quantitative determination of C-reactive protein in serum, the level of which is less than 20 mg/l, which is in favor of viral infection.

Monoclonal Gammopathy Of Unknown Significance - A Common Incidental Finding - What is to be thought of? What has to be done?

Monoclonal gammopathy of unclear significance (MGUS) is detected at high frequency in a variety of disciplines as an incidental finding. MGUS can be associated with non-malignant diseases, but it also can be a precursor of malignant lymphoproliferative disorders (multiple myeloma, Waldenstroem's disease, other non-Hodgkin's lymphoma, light chain (AL) - amyloidosis). However, many of these patients remain asymptomatic throughout their lives. Screening is performed by serum protein electrophoresis, immunofixation and determination of quantitative immunoglobulins and free light chains in serum. Currently, general population screening is not recommended. There are three subtypes with different rates and types of progression: IgM-MGUS, non-IgM-MGUS, and light-chain MGUS. The scope of further diagnostics and follow-up is based on the clinical findings and risk stratification (monoclonal protein in serum < or ≥ 15 g/l and normal or abnormal free light chain ratio in serum). If paraprotein-associated disease is detected, gammopathy is of clinical significance and should not be referred to as MGUS.

Acute behavioural and physiological responses of sheep to S/C ivermectin injection

Acute behavioural and physiological responses of sheep to S/C ivermectin injection

Effect of nutritional status of pregnant women on birth weight of newborns at Butajira Referral Hospital, Butajira, Ethiopia.

Back ground:Maternal nutritional status influences the developmental environment of the fetus which consequently affects the birth weight of the newborn. However, the association between maternal nutritional factors and birth weight is complex and is not well characterized in Ethiopia.Objective:To assess the effect of maternal anthropometry and biochemical profile on birth weight of babies at Butajira Referral Hospital, Butajira, Ethiopia.Methods and materials:Laboratory-based cross-sectional study was conducted among 337 pregnant women at the hospital. Socio-demographic and obstetric characteristics were collected using pre-tested questionnaires. Blood sample was collected from each pregnant women for determination of total serum protein, total serum cholesterol and hemoglobin level. However, maternal dietary habits were not assessed in this study.Results:A total of 337 pregnant women were involved in the study. The mean (standard deviation) birth weight of the newborns was 3.14 ± 0.46 kg. After adjusting for different maternal factors, parity (p = 0.013), hemoglobin level (p = 0.046), pre-pregnancy body mass index (p < 0.001) and weight gain during pregnancy (p < 0.001) were positively associated with birth weight of the newborns, while the associations with total protein (p = 0.822) and total cholesterol (p = 0.423) were not significant.Conclusion:This study has shown that nutritional status of pregnant women as indicated by maternal anthropometry and hemoglobin level was associated with birth weight of the baby. Therefore, nutritional status of the pregnant women should be improved to reduce the risk of low birth weight.

The indices of metabolismin rats with periodontitis on the background of hyper- and hypothyroidism

Introduction. The multifaceted action of thyroid hormones on the rate of metabolic processes reflected on the tissues of periodontium has been known for a long time. In this case, the prevalence of periodontitis in the population, the complexity of the implementation of prevention and treatment of the disease, ambiguity in the interpretation of the main pathogenetic mechanisms makes this problem extremely relevant.The aim of the study – to learn the indices of metabolismin blood serum of rats with periodontitis without concomitant pathology and against the background of hyper- and hypothyroidism.Research Methods. The study was carried out on 48 white non-linear male rats. Determination of glucose, total protein, albumin, urea, creatinine, total cholesterol, α-cholesterol and β-cholesterol in serum was carried out on a semi-automatic biochemical analyzer Humalyzer 2000 (Human, Germany).Results and Discussion. Experimental periodontitis is accompanied by a decrease in the concentration of glucose in the blood serum, hypoalbuminemia, an increase in the concentration of urea, total cholesterol, β-cholesterol and a decrease in the concentration of α-cholesterol. The imbalance of thyroid hormones has a multifaceted effect on metabolism in experimental periodontitis. Thus, in rats with periodontitis in case of hyperthyroidism, the concentration of glucose and total protein increased, albumin concentration decreased, urea concentration increased, total cholesterol and α-cholesterol concentrations decreased, and no significant changes in β-cholesterol concentration were observed. In rats with periodontitis, combined with hypothyroidism, glucose concentration decreased, total protein concentration increased, albumin concentration decreased, urea concentration increased, total cholesterol and β-cholesterol concentrations increased, and the concentration of α-cholesterol decreased.Conclusions. Reasonable changes in all major spectra of metabolism – carbohydrate, protein and lipid in case of experimental periodontitis indicate that periodontitis is a local process that affects the general state of the organism. The imbalance of thyroid hormones has a pronounced effect on the overall metabolic processes, especially on proatherogenic changes in serum, which are more pronounced in case of an experimental periodontitis in the background of hypothyroidism.

The electrophoretic pattern of serum proteins in dairy cows with inflammatory diseases

AbstractThe objective of the study was to evaluate the electrophoretic pattern of serum proteins in dairy cows suffering from various inflammatory diseases, and to study the influence of these diseases on the concentrations of protein fractions. Fourty dairy cows with clinical signs of various inflammatory diseases were used in the study, including cows withpost-partummetritis (n=10), mastitis (n=7), and hoof diseases (n=23). The cows were of a black pied Holstein-Friesian breed, Slovak spotted breed and their crossbreeds at the age of 3.5 to 8 years. Fourteen clinically healthy dairy cows were taken as the control group. The cows were blood sampled for the determination of total serum proteins and serum protein fractions. The protein fractions were divided into albumin, α1-, α2-, β1-, β2-, and γ-globulins. In cows withpost-partummetritis we found significantly lower concentrations of albumin (P&lt;0.001) and significantly higher values of α1-globulins (P&lt;0.01) compared with healthy animals. Significantly higher concentrations of β1-globulins were observed in cows with mastitis (P&lt;0.001), while the γ-globulin fraction was non-significantly higher. In cows with hoof diseases we recorded significantly lower values of albumin (P&lt;0.001), significantly higher concentrations of α1- and β1-globulins (P&lt;0.001), and non-significantly higher β2- and γ-globulins. Moreover, in the electrophoretic pattern of serum proteins we found β-γ bridging in thirteen cows with hoof diseases. Presented data suggest marked influence of inflammatory diseases on the concentrations of serum protein fractions in dairy cows. However, further investigations are needed to establish the diagnostic utility of serum protein electrophoresis in bovine clinical practice in relation to inflammatory diseases.

Effect of Egyptian propolis on cryptosporidiosis in immunosuppressed rats with special emphasis on oocysts shedding, leukogram, protein profile and ileum histopathology.

To investigate the activity of Egyptian propolis extracts (ethanol and water) on cryptosporidiosis in experimentally infected dexamethasone-immunosuppressed rats. A total of 180 male rats (190-220) g BWt were randomly divided into 9 equal groups (G1-G9). Groups of rats were kept as (G1): normal control, (G2-G9): immunosuppressed with dexamethasone and (G3-G9): infected with Cryptosporidium oocysts. Rats from (G4-G9) were given orally ethanol and water extract of propolis (at a dose of 50mg/kg BWt) and nitazoxanide (standard anti-cryptosporidial drug at a dose of 100mg/kg BWt) to infected rats with different regimes. Faecal pellets were collected from all groups to monitor oocysts shedding from the 2nd to the 15th day post infection. At the end of the experiment, blood was collected from all groups for determination of leukogram and serum proteins. Ileum specimens were also examined histopathologically. The highest reduction of oocysts shedding in faecal samples was 88% in rats prophylactically treated with propolis ethanol extract at the 4th dpi, and in rats prophylactically treated with water extract of propolis, was 91% at the 6th dpi. There was a marked increase in neutrophils count and α2- and β-globulins levels in infected rats treated with both extracts, while a significant decrease was detected in lymphocytes compared to the infected non treated group. β-Globulin level markedly increased in the rats administered nitazoxanide. Histopathological changes were observed in the ileum of rats infected with Cryptosporidium. Egyptian propolis extracts have an activity on cryptosporidiosis in rats. Moreover, propolis modulated the immunity in dexamethasone-immunosuppressed rats.

Innovative gold-free carbon nanotube/chitosan-based competitive immunosensor for determination of HIV-related p24 capsid protein in serum

The first competitive immunosensor for the determination of HIV-related p24 capsid protein, realized and validated in undiluted human serum.

Serum proteins and their diagnostic utility in veterinary medicine: a review

The evaluation of serum proteins and their electrophoretic pattern is a well-established laboratory method in the diagnosis of many diseases in humans, which has replaced the biochemical determination of the concentrations of albumin and the ratio of albumin to globulins. The measurement of serum proteins may be an important diagnostic tool for the detection, diagnosis, and monitoring of various diseases and pathological processes. The results of serum protein electrophoresis can be one of the most useful diagnostic aids in a wide spectrum of diseases, including infectious and inflammatory diseases, renal or gastrointestinal disorders, immunodeficiency states, as well as paraproteinaemias caused by lymphoid or plasma cell neoplasia. Although many studies have been carried out to determine the usefulness of the determination of serum proteins and their electrophoretic pattern in various disease conditions and disorders also in animals, serum protein evaluation is still a relatively little-used diagnostic tool in veterinary medicine. In this article, methods of serum protein determination, their diagnostic utility in animal care practice and their different patterns in dysproteinaemias and paraproteinaemias are reviewed.

Determination of Proteins in Human Serum by Near-Infrared Spectroscopy with Partial Least Square Analysis

ABSTRACTNear-infrared spectroscopy with interval and synergy interval partial least squares were used to determine total proteins, albumin, and globulin in human serum. Quantitative models of the relationships between the near-infrared spectra and these proteins were established. The root mean square error of prediction (RMSEP) was used to evaluate the models. The results demonstrate that synergy interval partial least squares was superior to interval partial least squares. The optimum wavelength ranges using synergy interval partial least squares for total proteins, albumin, and globulin in serum were 1668–1772 and 1772–1874 nm; 812–1000 and 1636–1800 nm; and 1228–1364 and 1642–1772 nm, respectively. The RMSEP of these proteins were 0.6905, 0.6686, and 0.5355 g/L. Waveband optimization was necessary to enhance the accuracy. The synergy interval partial least square model for total proteins, albumin, and globulin in serum offered higher accuracy and overcomes shortcomings of interval partial least squares. Furthermore, the optimized spectrum region may allow the design of near-infrared spectrophotometers for specific applications.