Enzyme Immunoassay Method Research Articles

Effect of Concomitant Chronic Obstructive Pulmonary Disease on Markers of Collagen Metabolism in Patients with Ischemic Heart Disease

To study parameters of collagen myocardial metabolism in patients with ischemic cardiomyopathy (ICMP) and to assess the effect of concomitant chronic obstructive pulmonary disease (COPD) on their levels. We examined 130 patients with ICMP (mean age 55.5 [43; 63] years) and 42 patients with ICMP and COPD (mean age 54.8 [41; 63] years). Determination of serum levels of matrix metalloproteinase-1 (MMP-1), its tissue inhibitor (TIMP-1), transforming growth factor-β1 (TGF-β1), and autoantibodies to collagen types I and III was carried out with the help of commercial test systems based on the method of solidphase enzyme immunoassay. Calculation of the volume fraction of interstitial collagen (VFIC) of the myocardium was carried out by the method of J. Shirani. In patients with ICMP combined with COPD, as well as in patients with ICMP we found disbalance in the metabolism of collagen of the extracellular matrix. At the same time, in patients with ICMP+COPD, processes of collagen degradation at the background of extracellular matrix fibrosis were found to be even more pronounced than in patients with ICMP. This was evidenced by the absence of statistically significant differences in the myocardial VFIC (p=0.703), and the level of TGF-β1 (p=0.074), as well as statistically significant higher activity of MMP-1 (p=0.037), MMP-1 / TIMP-1 ratio (p=0.045), and production of autoantibodies to collagen types I and III (p=0.042, p=0.039, respectively). The presence of COPD in patients with ICMP is associated with increased disbalance of the proteolysis-antiproteolysis system in the direction of collagen breakdown that can lead to more pronounced structural changes of the myocardium, and, as consequence, to aggravate functional impairment of the organ.

Rumination time as an indicator of stress in the first thirty days after calving.

The objectives of this study were to examine the option of being able to use rumination time (RT) as a form of stress indicator in the first thirty days after calving, and to determine the relationship between rumination time, blood cortisol levels, and lactate concentration levels in dairy cows during the first thirty days after calving. Ninety cows which produced milk (DIM) within 1-30 days were selected and categorised into the following groups: the first group (1) fell within 1-7 days after parturition (dpp) (n=30); the second group (2) fell within 8-14dpp (n=30); and the third group (3) fell within 15-30dpp (n=30) after calving. The cows were milked using Lely Astronaut® A3 milking robots with free traffic. The blood samples were tested using the fluorescence enzyme immunoassay method for cortisol analysis. Lactate concentrations were tested with a Lactate Pro2 ®. The RT increased during all of the exploratory periods (with readings between 1.12-4.90%). A decrease was also observed in the lactate levels (by 1.10 times) and cortisol levels (by 1.98 times, p⟨0.05) of cows which fell within the 8-14dpp group, when compared to an average of 1-7dpp in the previous study period (15-30dpp). However, lactate concentrations increased (by 1.84 times, p⟨0.05) as well as cortisol levels (by 2.09 times, p ⟨0.01) when compared with a figure between 8-14 dpp on the average. The results obtained indicate that, RT increased during all exploratory periods, while a decrease by 1.10 times and 1.98 times was observed in lactate levels and cortisol levels, respectively. During the entire period of the study RT was positively correlated with the lactate concentration levels, and negatively correlated with cortisol levels. Within a period of 1-14 days, a negative correlation was determined with lactate levels along with a 15-30dpp-positive correlation coefficient. In conclusion, RT can be used as a kind of stress indicator for cows in the first thirty days after calving; however, further research is required to ascertain this conclusion.

Comparison study between two enzyme immunoassay methods for the determination of mycophenolate acid

Comparison study between two enzyme immunoassay methods for the determination of mycophenolate acid

Relationship between insulin-like growth factor-1 (IGF-1) concentrations and body trait measurements and climatic factors in prepubertal goat kids.

This study aimed to investigate relations between insulin-like growth factor-1 (IGF-1) concentrations and some body trait measurements (body weight, withers height, rump height, body length, chest depth, chest width, chest girth and cannon bone circumference) and climatic factors in prepubertal male and female White (75 % Saanen and 25 % Kilis goat) and Angora goat kids. For this purpose, blood samples were regularly taken from the vena jugularis, and body trait measurements were regularly carried out (every 15 d for 5 months) on each kid. The IGF-1 analysis on the blood serum was performed using the enzyme immunoassay (EIA) method. Climatic values and the length of the photoperiod were obtained from the Turkish State Meteorological Service for the experimental period, and the temperature–humidity index (THI) was calculated using these values. Statistical analysis showed that the IGF-1 concentrations were higher () in female White goat kids. Furthermore, differences in IGF-1 concentrations were found () between periods and between the gender groups for both the White and the Angora goat kids. Moreover, the difference between the IGF-1 concentrations between genders was higher () in White goat kids. Additionally, positive and significant correlations were found between IGF-1 concentrations and some body trait measurements in prepubertal kids, except for in female White goat kids. In summary, it was found that there was a significant relationship between IGF-1 concentrations and growth characteristics of the goat kids. Furthermore, IGF-1 concentrations in the goat kids were significantly influenced by climatic factors such as photoperiod, temperature and the temperature–humidity index, with the release of IGF-1 increasing due to increases in the photoperiod and the environmental temperature.

THE EFFECTIVENESS OF IMMUNOVAC VP-4 FOR IMMUNOLOGICAL PARAMETERS IN FREQUENTLY AND LONG-TERM ILL CHILDREN

Aim. To study the level of specific antibodies of different isotypes to the antigens of Staphylococcus aureus and Klebsiella рneumoniae in the serum, saliva and nazal secret and the concentration of IgA,sIgA,IgG in saliva from frequently and long-term ill children in nasal-oral administration of Immunovac VP-4. Materials and methods. Specific antibodies to S. aureus and K.pneumoniae, contained in saliva, nasal, and serum were determined by the method of enzyme immunoassay. Concentrations of immunoglobulins of classes G, A and sА in saliva were determined by radial immunodiffusion using a commercial kit produced by the NPC «Medical immunology». Results. The high level of specific antibodies contained in the serum and nasal secretions, the level of antibodies in saliva is negligible. The serum is dominated by IgG-isotype antibodies, saliva and nasal secret — antibodies of IgA-isotope. After the introduction of Immunovac VP-4 there was a statistically significant increase in the level of specific antibacterial antibodies in serum, saliva and nasal secret, and increasing levels of IgG and sIgA in saliva. Conclusion. Obtained data demonstratet that the nasal-oral scheme of administration of Immunovac VP-4 frequently and long-term ill children allowed to increase the level of specific antibodies in serum, saliva and nasal secret to bacterial antigens that are part of the vaccine and to normalize the local synthesis of IgG and sIgA, which play a major role in the protection of the respiratory tract and mucous membranes of the upper respiratory tract.

Protein Induced by Vitamin K Absence II (PIVKA-II) as a potential serological biomarker in pancreatic cancer: a pilot study.

IntroductionProtein induced by vitamin K absence II (PIVKA-II) is an abnormal prothrombin increased in gastrointestinal malignancy. We aimed to evaluate PIVKA-II in comparison to established pancreatic cancer (PC) biomarkers (CA 19-9, carcinoembryonic antigen (CEA) and CA 242) measured in PC patients and in patients with benign pancreatic diseases.Materials and methodsWe studied 26 PC patients (Group 1) and 20 patients with benign pancreatic diseases (Group 2). PIVKA-II and CEA were measured by chemiluminescent enzyme immunoassay method (CLEIA) on LUMIPULSE G1200 (Fujirebio-Europe, Gent, Belgium), CA 19-9 and CA 242 were measured by ELSA (CisBio Bioassays, Codolet, France) and EIA (Fujirebio Diagnostics AB, Göteborg, Sweden), respectively. Receiver operating characteristic (ROC) analysis was performed to assess biomarkers’ diagnostic characteristics in both groups.ResultsMedian and interquartile range (IQR) in Group 1 and Group 2 were: 1749.0 (320.2 – 3921.0) vs. 31.0 (23.0 – 43.0) mAU/mL (P < 0.001) for PIVKA-II, 260.0 (158.7 – 272.0) vs. 45.2 (9.0 – 58.0) U/mL (P = 0.034) for CA 19-9, 104.0 (30.2 – 150.0) vs. 7.2 (4.8 – 26.0) U/mL (P < 0.050) for CA 242, 9.4 (5.3 – 37.5) vs. 4.5 (1.8 – 7.0) ng/mL (P = 0.021) for CEA. Areas under the ROC curve of PIVKA-II, CA 19-9, CA 242, CEA were 0.86 (95% CI: 0.71 – 1.00), 0.58 (95% CI: 0.38 – 0.78), 0.73 (95% CI: 0.54 – 0.92), 0.64 (95% CI: 0.44 – 0.85), respectively.ConclusionsPIVKA-II is significantly higher in PC than in benign pancreatic diseases. PIVKA-II shows a rather good diagnostic performance compared to CA 19-9, CEA and CA242, thus its determination could help PC management.

Seroprevalence of hepatitis B, hepatitis C, and HIV in pregnant women from Eastern Turkey.

The vertical transmission of hepatitis B virus, hepatitis C virus (HCV), and human immunodeficiency virus (HIV) infections are essential public health problems. In this study, we aimed to investigate the seroprevalence of the aforementioned infections among pregnant women. This study was done retrospectively on pregnant women who presented for antenatal follow-up and delivery between 2013 and 2016. Data were collected from the hospital's electronic health records and patient files. Blood samples were analyzed at the microbiology laboratory of the hospital. HBsAg, anti-HBs, anti-HCV, and anti-HIV titers were tested using the chemiluminescence enzyme immunoassay method (Architect, Abbott Laboratories, USA). HBsAg and anti-HBs levels were tested in 35,295 pregnant women aged 18-45 years. The HBsAg and anti-HBs levels were positive in 425 (1.2%) and 9583 (27.7%) patients, respectively. From 2013 to 2016, the HBV carrier rates have continuously decreased from 1.4% to 0.8%, whereas the anti-HBs positivity has increased from 25.4% to 30.2%. Anti-HCV was detected in 6 of the 9709 (0.06%) patients. All the 7113 pregnant women screened for HIV showed negative results. Hepatitis B carrier rates among pregnant women gradually decreased with a simultaneous increase in the immunity rates. HCV seroprevalence was low and HIV positivity was not encountered in the study population.

Exposures to L‐felinine Suppressed Plasma Testosterone in Laboratory Rats

Felinine is a unique sulfur‐containing amino acid found in the urine of domestic cats and select members of the Felidae family (Rutherfurd et al. 2002). L‐felinine and its sulfur‐containing volatile derivatives excreted with urine in sex‐ and age‐dependent manner (Miyazaki et al. 2006). These cat‐specific volatile compounds may represent pheromones used as territorial markers for conspecific recognition or reproductive purposes by mature cats (Miyazaki et al. 2008). Species‐specific compounds may be used also by other species to recognize potential predators and their physiological status. In our previous studies we showed that exposures of rats from laboratory heterogenous population during gestation to the cat urine or L‐felinine significantly affected reproductive output. We observed significantly lower weight of newborn pups and skewed sex ratio in favour in males. At the behavioral level, exposed to cat urine/L‐felinine male rats showed fewer mountings and mountings with intromissions in presence of receptive females. In the current study we examined the influence of domestic cat‐specific compound L‐felinine on testosterone secretion in male rats. Intact cat urine was used as positive control; tap water was used as negative control. Test subjects were 3–4 month old Norway rats Rattus norvegicus from laboratory population. Males were placed singly two weeks prior to experiment. We used concentration of L‐felinine comparable to naturally occurring in the adult cat urine (Rutherfurd et al. 2002). Cotton balls soaked with L‐Felinine (0.05% w/v, 0.2 ml, US Biologicals), cat urine (0.2 ml) or tap water (0.2 ml) in plastic perforated containers were placed directly into home cages of rats. We renewed test solutions twice a week. Experimental animals were randomly assigned to one of treatment groups. Blood samples (50 μl) were obtained from rear foot pad (Miller et al. 1997). Plasma testosterone was assayed (in duplicate) by enzyme immunoassay method (EIA‐1559, DRG). Exposure of male rats to cat urine (0.2 ml) during two weeks significantly suppressed plasma testosterone (p=0. 0020, n=10, t‐test). Analogous exposures to L‐felinine (0.05 %, 0.2ml) produced quite similar effect (p =0.049, n=10, t‐test) in male rats. The effect was dose dependent: we did not observe suppression of plasma testosterone when lower amount of L‐felinine solution (0.05 %, 0.1ml.) was applied to home cage of male rats. It appears to be that altered sexual behavior of male rats exposed to L‐felinine/cat urine could be explained by significantly lowered testosterone secretion. The expression of the secondary defense reactions by laboratory naive rats and the failure tohabituate at the hormonal level may indicate an innate nature of the response.Support or Funding InformationSupported by Russian Foundation for Basic Research grant 19‐34‐70024 mol_a_mosThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

THE RESPONSE OF THE ORGANISM OF ADOLESCENTS TO THE INHALATION CHEMICAL LOAD AND INFESTATION WITH GIARDIA

Introduction. The anthropogenic load affects the human body, creating the prerequisites for the development of adaptation or maladaptation processes, in which the immune system plays a major role. Giardia infection also affects the immune system. The aim of the study was to assess the levels of cytokines and immunoglobulin A in adolescents with giardiasis living in conditions of the different inhalation chemical load Material and Methods. 319 adolescents were included in the study. The individual inhalation chemical load on the adolescents was calculated taking into account data on the content of impurities in the atmospheric air, indoor air, personalized data of schoolchildren. Three groups were selected in accordance with the individual values of the hazard index of the immunity disorders in students. Cases with and without Giardia infection were identified in each group. The content of antibodies to lamblia, interleukins-2 and -10, interferons-alpha and-gamma, immunoglobulin A in schoolchildren was studied by the method of enzyme immunoassay. Results. The chronic inhalation chemical load with immunotropic substances have been established to cause a decrease in the levels of INF-α, INF-γ, and IgA. Giardia was found to induce the increase in the levels of IgA, IL-2, INF-α, and INF-γ, at hazard indices of the development of pathology of the immune system less than 2. Discussion. Giardia invasion causes an increase in INF-α, INF-γ, IL-2, IgA levels, which indicates the activation of the immune protection when hazard indices of a danger of impaired immunity were less of 2. Differences in cytokine levels in adolescents with and without giardiasis are leveled when hazard indices were more than 3, which is probably the result of changes in the immune system due to a high level of the chemical stress. Conclusion. The presence of parasitic infestation and chemical air pollution have a synergistic inhibitory effect on the immune system, namely, they reduce IL-10, Ig A, INF-α, and INF-γ levels, with the higher inhalation load by substances tropic to the immune system

Platelet Aggregation and Interleukins Indicators Impacting the Outcomes of Ischemic Stroke

Aim: Relative assessment and relationship between of platelet aggregation and the level of interleukins IL-1β, IL-4, IL-6, and IL-18 among ischemic stroke (IS) patients. Methods: A prospective clinical cohort study involved 108 IS patients, classified into group 1—surviving (93; 86.1%) and group 2—lethal outcomes (15; 13.9%). The studies were conducted in the most acute (first day of hospitalization) and acute (7 days of hospitalization) phases of IS. The level of interleukins was defined by enzyme immunoassay method. The platelet aggregation research was performed by the nephelometric method. Results: In the acute phase of the IS the first group, patients showed reduction in the level of IL-1β and IL-6, and increase in the level of IL-4, compared to the most acute period, whereas in patients of the second group further increase in IL-1β and IL-6, and reduction of IL-4 levels were recorded. Both in the most acute and acute phases of the IS, in parallel to the aggravation of platelet aggregation (PA) disturbances, the lethal outcome probability grew. The latter also grew together with increase in the synthesis of IL-1β and IL-6, in the most acute phase of the IS. In the acute phase of the IS, lethal outcome was recorded in all patients with high values of IL-1β and IL-6, and all patients with low values of IL-1β showed improvement. Conclusions: Vivid disturbances of PA, IL-1β, IL-6, and IL-18 synthesis, in the most acute phase of the IS, signal higher probability of lethal outcome. Reduction in the levels of IL-1β, IL-6, and IL-18, and increase of IL-4 in the acute phase, means improvement in conditions.

Direct competitive enzyme immunoassay method for the determination of ochratoxin a in feeds and foods

A reagent kit EIA-OCHRATOXIN A for the determination of mycotoxin ochratoxin A in feeds and foods by a direct competitive enzyme immunoassay using microtitration plate has been developed and tested. The conjugate of ochratoxin A with horseradish peroxidase was synthesized using the mycotoxin aminoderivative joined to the carbohydrate chain of the enzyme. A monoclonal antibody to the mycotoxin was immobilized on the surface of plate wells through sheep anti-mouse antibodies. The evaluated parameters of the kit and metrological characteristics of the technique of measurements correspond to the modern level of immunoassay development and provide the determination of ochratoxin A content of agricultural products in a range of 5 to 375 mg/kg with proper accuracy and precision. The limit of quantitative determination of ochratoxin A in grain and cereal foods does not exceed 5 mg/kg.

Adipose-Derived Mesenchymal Stem Cells Can Ameliorate Airway Remodelling and Improve Oxidative Stress: An Open Label and Randomized Clinical Trial Study on Sulfur Mustard Exposed Patients

Background: Lung is a primary target for sulfur mustard (SM) toxicity. Here, we investigated the potential efficacy and safety of adipose derived mesenchymal stem cell (AdMSC) on respiratory quality, lung regeneration, oxidative stress, and airway remodelling in patients with mustard lung. Methods: In this clinical trial study, 10 patients received 100x106 cells every 20 days for 4 injections over 8 weeks. The pulmonary function tests (PFT), chronic obstructive pulmonary disease (COPD) Assessment Test (CAT), Borg Scale Dyspnea Assessment (BSDA), and 6 Minute Walk Test (6MWT) were evaluated. Total antioxidant capacity (TAC), malondialdehyde (MDA) and glutathione (GSH) were measured in the sputum of all samples. Changes in expression of metallothionein 3 (MT3), glutathione reductase (GR), glutathione peroxidase (GPX), and lacto peroxidase (LPO), as well as matrix metallopeptidase 2 (MMP2), matrix metallopeptidase 9 (MMP9), transforming growth factor-b1 (TGF-b), vascular endothelial growth factor (VEGF), metallopeptidase inhibitor 1 (TIMP1), and metallopeptidase inhibitor 2 (TIMP2) were assessed using real-time PCR. Clara cell protein 16 (CC16) and Mucin-1 protein (KL-6) were measured in the sputum all samples as biomarkers of lung injury using enzyme immunoassay method. This study is registered with ClinicalTrials.gov (NCT02749448). Findings: Spirometry test results were slightly improved after each injection. A gradual improvement was observed in the mean of 6MWT scores. CASA-Q and VAS scores were increased, while means of CAT and Borg-Score were decreased. A significant improvement was found for the mean of KL-6 (p=0.022) and CC16 (p=0.005) levels. Sputum levels of TAC and GSH had a tendency to be higher after each injection, while MDA content was decreased. An improvement was observed for antioxidants genes, especially for GSR, MT3 and LPO, compared to the baseline. A gradual upregulation was found for VEGF and TGF-β expression after MSCs therapy. MSCs caused a balance between MMP9/TIMP1 and MMP2/TIMP2 ratio. The MMP2/TIMP2 ratio was decreased from 4.9 on baseline to 0.71 on day 180 (p<0.001), while the MMP9/TIMP1 ratio was declined from 16.35 on baseline to 0.64 on day 180 (p<0.001). Interpretation: The findings suggest that MSCs therapy can be safely used for improvement of lung injury and regeneration in these patients without any adverse effects. Trial Registration Number: This study is registered with ClinicalTrials.gov (NCT02749448). Funding: Iran National Science Foundation and Veterans Foundation. Declaration of Interest: The authors confirm that there are no conflicts of interest. Ethical Approval: The ethics review board of the Baqiyatallah University of Medical Sciences (Cod: IR.BMSU.REC.1393.32) approved this survey and all patients signed an informed consent letter before the study.

Severe 25-OH vitamin D deficiency as a reason for adverse pregnancy outcomes

Objective: There is a growing concern about the unfavorable effects of vitamin D deficiency in general population, including pregnant women, worldwide. The aim of this study is to evaluate the effect of severe maternal serum 25-OH vitamin D levels on adverse pregnancy outcomes in first trimester.Material and methods: Serum samples of 86 pregnant women in first trimester were collected prospectively from May 2017 to June 2017. Serum 25-OH vitamin D levels were analyzed by enzyme immunoassay method. Patients were classified according to maternal serum 25-OH vitamin D levels as group 1 (n = 34) <10 ng/ml and group 2 (n = 52) >10 ng/ml. The two groups were compared in terms of adverse pregnancy outcomes.Results: The mean 25-OH vitamin D levels of the total 86 pregnant women were 13.83 (6–48) ng/ml. 40% of the pregnant women had low level of 25-OH vitamin D levels (<10 ng/ml). The adverse pregnancy outcomes were significantly increased in group 1 (p<.018).Conclusions: Maternal serum 25-OH vitamin D levels <10 ng/ml is a risk factor for adverse pregnancy outcomes. 25-OH vitamin D levels should be screened in high-risk pregnant women and treated in case of deficiency.

Performance of Metabolic Parameters in Formation of Sleep Disorders in Menopausal Women

Background. It is known that menopausal women suffer from sleep disorders. As additional diagnostic methods, it is possible to use the equations of linear classification functions.Aim: to determine the most informative parameters among the lipid profile, the “lipid peroxidation – antioxidant protection” system parameters and the melatonin secretion chronobiological rhythms in menopausal women with sleep disorders.Materials and methods. 56 perimenopausal women and 70 postmenopausal ones in accordance with the inclusion criteria participated in this study. Diagnoses of insomnia and obstructive sleep apnea syndrome were made according to the results of specialized somnological questionnaires and polysomnographic monitoring. Spectrophotometric methods for studying of lipid metabolism and the “lipid peroxidation – antioxidant protection” system parameters, an enzyme immunoassay method for determining melatonin were also used. A multidimensional discriminant analysis to reveal the most informative parameters among the parameters studied was used.Results. The most informative indicators of the metabolic system for sleep disorders in perimenopausal women with insomnia are melatonin 06.00–07.00 h, melatonin 18.00–19.00 h, melatonin 23.00–00.00 h, ketodienes and conjugated trienes, oxidized glutathione; with insomnia and OSAS – melatonin 06.00–07.00 h, melatonin 12.00–13.00 h, melatonin 23.00–00.00 h, substrates with conjugated double bonds, total cholesterol; in postmenopausal women with insomnia – diene conjugates, α-tocopherol, total antioxidant capacity, active products of thiobarbituric acid, reduced glutathione; with insomnia and OSAS – total antioxidant capacity, α-tocopherol, substrates with conjugated double bonds.Conclusions. The results obtained indicate a great strain in the antioxidant protection system in menopausal women with sleep disorders in response to changes in free radical homeostasis.

A Study to Assess the Significance of Urinary Collagen IV Excretion to Predict Preeclampsia in Early Pregnancy

It was a prospective study with a case control design. The purpose of the study was to measure the urinary collagens IV (U-coll IV) and microalbumin (MA) level in early pregnancy and to explore the role of excretion U-coll IV in prediction of preeclampsia (PE). A total number of 119 pregnant women at 10-14 weeks of pregnancy were selected on the basis of availability. MA by immunoturbidimetry assay and U-coll IV by enzyme immunoassay method were measured in these subjects and they were followed up to the term for the possible development of PE. MA was defined by albumin-creatinine ratio (ACR) above 32mg/g and high U-coll IV was defined by values above the cut-off point 2.74 ng/ml determined by the median value of the (controls). The data were analyzed by grouping the subjects who developed PE in later stages of pregnancy (the PE group) and those who did not develop PE in later stages (Control group). From the total subjects, 10 developed PE which shows a prevalence of about 8.4%. The PE group had higher value of ACR as compared to Control [ACR, mg/g, median (range), 42.3]. The sensitivity of ACR in predicting PE was 80%, specificity 49.54%, PPV 12.69% and NPV 96.42% respectively. The sensitivity of high U-coll IV in predicting PE was 70%, specificity 50.5%, PPV 11.5% and NPV 94.8%. It may be concluded that early pregnancy levels of MA and U-coll IV can be used as predictors of PE with high negative predictive value; and U-coll IV has no added advantage over MA in this respect.

Detection of Cortisol Level via Enzyme Immunoassay Influenced by Exercise

Cortisol is among the human body’s most necessary hormones. On any given day, cortisol’s primary function is to control how our body creates and utilizes energy. This study examined the influence of exercise on the cortisol levels.Twenty female subjects had their cortisol levels determined before their morning exercise (Pre) and after their morning exercise (Post) using microplate enzyme immunoassay method. The result of study showed that there was significant difference (P&lt;0.05) in the cortisol levels before exercise 13.49 ± 0.90µg/dl Compared with after exercise 15.10±1.08µg/dl. It was also observed that there was no significant difference in the cortisol level (P&gt;0.05) obtained from the female subjects before and after exercise between the ages of 21-23, 24- 26 and 27-29 years. Collectively, these results showed that exercise provoked increased in cortisol level. Keyword: Cortisol, Immunoassay, Exercise, Hormone

INVESTIGATION OF PREBIOTIC, IMMUNOSTIMULATING PROPERTIES OF FUCOSE AND ITS EFFECT ON REPRODUCTIVE FUNCTION

Aim. Investigation of the effect of fucose in the diet on the gastrointestinal microflora of experimental animals with experimental dysbiosis, the humoral factors of nonspecific immunity, as well as the degree of fucosylation of oocytes and the proportion of oocytes that can be fertilized. Materials and methods. Prebiotic properties of fucose were studied by analyzing the luminal microflora of experimental mice against the background of experimental dysbiosis. Investigation of factors of nonspecific immunity was carried out after immunization of mice according to the level of antibody formation in blood serum by the method of enzyme immunoassay. The degree of fucosylation of oocytes was assessed by the intensity of their luminescence upon microscopy of oocytes of experimental mice on a fluorescent microscope. Results. The use of fucose in all tested doses led to the restoration of the composition and quantity of the gastrointestinal microflora. For the correction of dysbiosis, the optimal concentration of fucose was 0.02% of the body weight of the experimental animals. Inclusion of fucose in a diet of experimental animals in the amount of 0.008% to the body weight provided the highest level of immune response. The degree of fucosylation of oocytes, the proportion of oocytes capable of fertilization was increased when fucose were introduced in the amount of 0.008% to the body weight of the mice. Conclusion. Bifidogenic and lactogenic activity of fucose is established. The ability of fucose to stimulate an increase in the level of antibodies in in blood serum is shown. The tendency of positive effect of fucose in the diet of mice on the degree of fucosylation of oocytes was revealed.

Peculiarities of Changing Bioregulators and Organic Acids in the Serum of Blood and Amniothic Fluid of Women with Spontaneous Preterm Labor

Background: Premature birth is currently a serious problem for obstetrician-gynecologists. Most researcher consider this problem from the standpoint of metabolome disturbances.Aims: determination of the level of bioactive regulators and organic acids with physiological and complicated pregnancy, ending in timely and spontaneous preterm labor, in blood serum and amniotic fluid and revealing the presence of their relationship for the development of premature birth and the condition of newborns.Materials and methods: 72 women were examined, 40 of them with a physiological pregnancy (control group) and 32 with premature labor (the main group). All studies were carried out in the blood serum and amniotic fluid of women of these groups. The content of TGFβ was determined by the method of solid-phase enzyme immunoassay. Identification and quantitative determination of citric and succinic acids were carried out by the method of capillary electrophoresis.Results: TNFα production in the blood serum of women with preterm labor, decreased by 38%, and the level of citric and succinic acids by 33 and 30%, respectively, compared with the control group. In the amniotic fluid the level of TNFα was increased 2.5 times and TGFβ ― 2.2 times, citric acid ― 79%.Conclusions: The revealed imbalance in the level of cytokines and organic acids studied in that biological fluids can evidently affect the fetal condition, which can be considered as possible markers of perinatal injuries and prognostic indicators of the state of the newborn.

Morphine Concentrations in Human Urine Following Poppy Seed Paste Consumption

Papaver somniferum (opium poppy) is one of the world’s oldest medicinal plants which are widely used for medicinal, nutritive and scientific purposes. Turkey is one of the major legal opium poppy producer countries in the world and the seed paste of the poppies is consumed in great deal, even more than 100g per meal. The main objective of this study is to investigate the influence of poppy seed paste ingestion on urine tests for opiates whether or not could lead to opiate positive urine test results. For this purpose, a variety of poppies were used and the morphine content of white, yellow and blue-black poppies were determined as 1.9, 4.0 and 2.6mg/kg, respectively. 100g of these seed pastes were consumed in the breakfast by ten healthy adults enrolled in the study over three days and urine samples were collected before and after the breakfast. Opiate screening analysis was carried out by enzyme immunoassay method and the results were evaluated by two different cut-off values (300 and 2000ng/mL). Morphine confirmation analysis was made by GC–MS system and the chromatographic method was validated in terms of selectivity, extraction efficiency, linearity (25–2000ng/ml), intra-assay precision, accuracy, limit of detection (LOD) and limit of quantitation (LOQ) (3 and 10ng/ml), carryover, matrix effect, dilution integrity and stability. According to cut-off value 300ng/ml, opiate concentrations were found positive up to 48h. For cut-off value 2000ng/mL; this time was up to 12h in collected urine samples after consumption of three different colored poppy seed pastes. In all urine samples, thebaine was detected while the heroin abuse metabolite 6-acetyl morphine (6-AM) was not. Urine drug testing legislation was revised on 2016 in Turkey and opiate screening cut-off values increased from 300 to 2000ng/mL. Overall results have shown that poppy seed paste as food consumption could lead to opiate positive urine test result even if increased cut off levels are used. It can also be deduced that thebaine can be taken as supportive biomarker for poppy seed paste consumption. Awareness of interpretation of urine test results and defining the procedures especially for forensic drug testing must be done in legal aspect to ensure justice for each individual (workplace, traffic, court etc.).

L-Carnitine Supplementation Increases Trimethylamine-N-Oxide but not Markers of Atherosclerosis in Healthy Aged Women

Background: L-carnitine can be metabolized to trimethylamine N-oxide (TMAO), a molecule that promotes atherogenesis through its interaction with macrophages and lipid metabolism. Objective: The aim of the present study was to assess whether L-carnitine supplementation may promote changes in selected serum biomarkers of atherosclerosis. Methods: Before the start, in the mid-point and after completing the 24-weeks supplementation protocol, fasting blood samples were taken from the antecubital vein. Plasma free L-carnitine and TMAO were determined by the UPLC/MS/MS method. Serum proteins were determined by the enzyme immunoassay method using commercially available kits. Total cholesterol, high-density lipoprotein-cholesterol, low-density lipoprotein-cholesterol, and triglycerides have been determined using standard automatic analyzer. Results: L-carnitine supplementation elevated fasting plasma carnitine in the mid-point of our study and it remained increased until the end of supplementation period. Moreover, it induced tenfold increase in plasma TMAO concentration but did not affect serum C-reactive protein, interleukin-6, tumour necrosis factor-α, L-selectin, P-selectin, vascular cell adhesion molecule-1, intercellular adhesion molecule-1 or lipid profile markers. Conclusion: We demonstrated that ­although oral L-carnitine supplementation significantly ­increased plasma TMAO concentration, no lipid profile changes or other markers of adverse cardiovascular events were detected in healthy aged women over the period of 24 weeks.