EIA Kit Research Articles

Comparison of Cortisol Levels in the Hair of Male European Roe Deer at the Beginning and End of the Stalking Hunting Season.

Roe deer (Capreolus capreolus), as a representative of the Cervidae family, are particularly sensitive to negative environmental stimuli due to their need to maintain increased vigilance during feeding. Intensive hunting seasons are undoubtedly also a stressful factor for this species. The aim of this study was to analyze the cortisol levels in the hair of male roe deer collected at the beginning and the end of the hunting season in eastern Poland. In total, hair samples from 26 individuals collected in May and 31 individuals collected in September were analyzed. The concentration of cortisol in the hair was determined using the EIA kit. It was shown that the concentration of cortisol was significantly higher in samples collected in September compared to those collected in May (Pr. > |t| = 0.0017). Moreover, the age of animals and carcass mass did not significantly affect the concentration of the tested hormone. In summary, the concentration of cortisol in the fur of male roe deer depended on the season and was not influenced by the carcass mass or age of the animals studied.

Endogenous Cannabinoids in Crohn’s Disease

Background: An increasing number of people around the world suffer from Crohn’s disease (CD), one of the inflammatory bowel diseases (IBDs). Recent evidence suggests that the endogenous cannabinoid system plays an important role in IBD. The main endocannabinoids, anandamide (AEA) and 2-arachidonoylglycerol (2-AG), are ligands of the CB1 and CB2 receptors that are expressed on immune cells. These receptors as well as endocannabinoids are believed to mediate inflammatory activity and participate in the pathophysiology of CD. Objective: Very little is known about the secretion of endogenous cannabinoids throughout the course of CD. Therefore, the aim of this study was to determine and analyze the 2-AG and AEA concentrations in children who suffered from CD. Methods: We studied 34 adolescents with CD, mean age 13.5 years ± 2.0. Blood samples were collected three times in the active phase of the disease (during admission to hospital, before treatment), 2–4 weeks later, during the treatment and clinical improvement of the patients, and 3–6 months later, in the period of remission. The control group included 33 healthy teenagers of the same age who were examined once. In all patients, fasting blood samples were collected in the morning. 2AG and AEA concentrations in serum were measured using EIA kits (Abclonal, Woburn, MA, USA and ELK Biotechnology Co., Ltd. Wuhan, China, respectively). Results: The median 2-AG concentration in the study group was stable: acute phase 907.4 [379.3; 1300.5] pg/mL, during treatment 715.1 [416.7; 1302.5] pg/mL; remission 991.1 [381.1; 1652.5] pg/mL, and similar to the values observed in the control group 761.8 [504.3; 1497.0] pg/mL. No significant differences were found between the results obtained at all time points in the study group and compared to the control group (in all cases p > 0.44). In the case of AEA, in the study group before treatment, the median concentration was 2.63 [2.24; 2.79] ng/mL and similar to values obtained during treatment 2.56 [2.33; 3.06] ng/mL, and in remission, with 2.61 [2.46; 2.85] ng/mL. All these concentrations were lower compared to the values measured in the control group of 3.18 [2.57; 3.88] ng/mL (p = 0.023, p = 0.035, p = 0.056, respectively). Conclusions: While 2-AG levels remained stable throughout the disease course and were comparable to controls, AEA concentrations were consistently lower in CD patients. Therefore, therapeutic interventions aimed at increasing AEA-related signaling, such as administration of its analogues or fatty acid amide hydrolase inhibitors, may improve symptoms associated with Crohn’s disease in children.

Genetic factors related to aspirin resistance using the Multiplate® device in Hong Kong Chinese patients with stable coronary heart disease

ObjectiveAssociations between single nucleotide polymorphisms (SNPs) and aspirin resistance (AR) have been studied with variable results. The associations of genetic variants with AR may be helpful to explain why some individuals demonstrate aspirin insensitivity with this anti-platelet therapy. The purpose of this research was to investigate the effect of different genotypes in candidate genes on aspirin response in patients taking long-term aspirin therapy by measuring the serum thromboxane B2 (TXB2) and platelet function using the Multiplate® analyser. MethodsA total of 266 patients with stable coronary heart disease (CHD) taking low-dose aspirin for long periods of time and without any other anti-platelet drugs medications were enrolled into the study. They were required to take 80 mg of aspirin every morning for a week including the day before blood tests. Blood samples were collected 24 h after the last dose. The 80 mg dose of aspirin was taken orally and blood samples were collected again 1 h later. The serum TXB2 levels were measured in samples at 24 h post-dose and 1 h post-dose using the EIA kit and platelet activity was determined using the Multiplate® Impedance Platelet Aggregometry (ASPI) assay. Genotyping assays were performed by the TaqMan SNP genotyping technique. ResultsOf the 266 patients, only 251 patients were enrolled in the present study. The PTGS1/COX1-1676 A > G (rs1330344) and the PTGS2/COX2-765 G > C (rs20417) SNPs showed significant associations with the ASPI measurements in samples taken at 24 h post-dose, but not with the values at 1 h post-dose or with the TXB2 levels (P < 0.05). ConclusionsOur results suggest that polymorphisms in the PTGS1/COX1 and the PTGS2/COX2 genes may be associated with reduced anti-aggregatory effects and increased the risk of AR, but future larger-scale cohort studies are necessary for further validation.

Relationships of apolipoprotein E genotypes with a cluster of seven in persons with type 2 diabetes.

The objective of the study was to determine if there would be statistically significant differences or trends among apolipoprotein E genotypes (2/2, 2/3, 2/4, 3/3, 3/4, and 4/4) for each member of the cluster of seven associated with type 2 diabetes (T2D). The cluster of seven includes abdominal obesity, hypertension, platelet hyperaggregability, hyperglycemia, dyslipidemia (decreased plasma levels of high-density lipoprotein cholesterol (HDL-C) and increased plasma levels of triglycerides)), increased low-density lipoprotein (LDL) oxidation, and increased inflammation. Forty-six patients with well-controlled T2D participated in the study. Abdominal obesity (assessed by waist circumference), hypertension (measured by manual sphygmomanometry), platelet hyperaggregability (measured by bleeding time), hyperglycemia (by enzymatic kit and spectrophotometry), decreased plasma levels of HDL-C and increased plasma levels of triglycerides (by enzymatic kit and spectrophotometry), increased LDL oxidation (measured by LDL conjugated dienes using spectrophotometry) and increased inflammation measured by C-reactive protein (CRP) (by EIA kit) were determined. All genotypes, except 2/2 were found in the population studied. Abdominal obesity did not vary significantly across the five genotypes. However, glucose levels trended progressively higher going from 2/3 to 2/4 to 3/4 to 4/4. Systolic blood pressure was higher in 3/4 compared to 2/4 and trended higher in 3/4 compared to 3/3. Diastolic blood pressure trended higher in 3/3 vs 2/4 and significantly higher in 3/4 compared to 2/4. Triglycerides trended higher in 3/4 vs 3/3 while HDL-C came close to trending downward in 4/4 compared to 2/4. Bleeding time was unaffected by genotype. Plasma LDL conjugated dienes trended higher in 3/4 vs 2/4 and were significantly higher in 3/4 vs 3/3. CRP trended higher in 4/4 vs 2/3. We can conclude that those with at least one 4 allele in the presence of another allele being 2, 3 or 4 is potentially (in the case of trends) deleterious or is deleterious in terms of hyperglycemia, hypertension (systolic and diastolic blood pressure), dyslipidemia, LDL conjugated dienes and CRP levels.

A Case Control Study Evaluating the Relationship between Vitamin K2 Serum Level and Periodontitis.

Vitamin K2 (VK2) is an essential co-factor for bone metabolism. There is still very little data regarding possible VK2 relation to periodontitis. This study aimed to investigate any potential link between VK2 serum level and the severity of periodontitis in comparison to a control group of healthy individuals. The trial was performed on 100 patients among whom 50 were diagnosed with periodontitis. The patients underwent full clinical periodontal and radiological examination. The VK2 serum level was assessed using the ELISA kit (Gla-type osteocalcin EIA Kit, Takara, Kusatsu). Patients with periodontitis had mean serum levels of VK2 significantly lower (0.27 ± 0.06 nmol/L; p < 0.001) than the control group (0.43 ± 0.09 nmol/L; p < 0.001) regardless of the patient's age or sex. The VK2 serum level decreased with the severity of periodontitis with the lowest level in stage IV of the disease (0.19 ± 0.01 nmol/L; p < 0.001). Also, a significant drop was noticed between the grades of periodontitis. Individuals with localized forms of the disease had significantly lower VK2 levels (0.26 ± 0.006 nmol/L; p < 0.001) in comparison to subjects with generalized periodontitis (0.30 ± 0.01 nmol/L; p < 0.001). The VK2 serum levels were also associated with most of the clinical parameters such as bleeding on probing (-0.805, 95% CI: -0.894 to -0.654, p < 0.001), attachment loss (-0.752, 95% CI: -0.862 to -0.574, p < 0.001), and bone loss (-0.656, 95% CI: -0.801 to -0.439, p < 0.001). In the present study, the VK2 serum level was correlated to periodontitis, and its severity, complexity, extension, and grade. The range of VK2 was decreasing together with the worsening of all clinical parameters of periodontitis.

10 Extra-label bisphosphonate effects on bone turnover biomarkers in juvenile, exercising horses

Bisphosphonates, such as clodronate disodium (CD), modulate bone metabolism and, though their extra-label use in young horses is common, impacts on skeletal development and longevity are unknown. The objective of the current study was to determine the effects of CD use on systemic biomarkers of bone turnover in juvenile, exercising horses. We hypothesized that bone resorption would decrease, and bone formation would increase as a result of CD administration and repeat treatments would have a greater effect. To test this, 32 juvenile Quarter Horses were used in a 168-d trial. Horses were stratified by age (500 ± 13 d), BW (336 ± 26 kg), sex (n = 16 geldings; n = 16 fillies), and initial bone optical density and randomly allocated to one of 4 treatment groups receiving either 1.8 mg/kg BW CD (OSPHOS®) or isovolumetric saline (placebo). Investigators were blinded to treatments which included a control (CON; n = 8), single-dose CD on d 84 (1X; n = 8), 2-doses CD on d 0 and 84 (2X; n = 8), and 4-doses CD on d 0, 42, 84, and 126 (4X; n = 8). Horses were housed individually in stalls (3.6 × 7.3 m), fed 1% BW/d concentrate, and allowed ad libitum access to Coastal bermudagrass hay and water. Horses exercised 5 d/wk using a free stall exerciser with a phase-based progressive workload; phase I (d 0–84) simulated sales preparation and phase II (d 85–168) mimicked an early training program. Blood was collectedon d 0, 42, 84, 126, and 168, before treatment injections on days of administration. Serum was analyzed for receptor activator of nuclear factor kB ligand (RANKL), tartrate resistant acid phosphatase 5b (TRAP5b), c-terminal crosslinks of type I collagen (CTX-1) and bone-specific alkaline phosphatase (BAP) via commercial ELISA or EIA kits. All data were analyzed using PROC MIXED of SAS with a baseline covariate for BAP and BAP:TRAP5b. No differences were seen in osteoclastic differentiation signal RANKL (P ≥ 0.24). In contrast, a treatment × time interaction was noted for osteoclastic TRAP5b (P = 0.03); specifically, TRAP5b decreased in 4X from d 0 to 126 before returning to baseline at d 168 and decreased in 2X at d 84 compared with baseline, while CON and 1X increased or remained the same over time. Concentrations of CTX-1, a type I collagen degradation marker, increased over time (P < 0.01) in all treatments. Likewise, osteoblastic BAP (P < 0.01) and BAP:TRAP5b (P = 0.04) increased from d 42 to 84 and remained elevated until d 168. These results indicate that multiple doses of CD (2X and 4X) over 168 d reduced activity of osteoclasts despite having no measurable effect on bone formation or type I collagen degradation.

8-isoprostane as a marker of oxidative stress in experimental animals after implantation of surgical mesh with a coating based on tantalum and its derivatives

Nowadays, the world observes the problem of adhesion process after implantation of polypropylene surgical meshes, which occurs in about 30–40 % of patients in the postoperative period. Over the past 30 years, tantalum and its derivatives have proven themselves as materials for biomedical implants in dentistry and orthopedics. In the future, tantalum-based coatings may become an inhibitor of the development of adhesion and inflammatory processes after surgical mesh implantation. In order to better predict the implantation of biomedical materials, it is important to determine the markers of the development of oxidative stress. A change in the concentration of biomarkers of the state of the oxidants system, in particular 8-isoprostane, can indicate the development of pathological conditions under the conditions of oxidative stress, which can have a significant impact on the viability of cells after surgical intervention. The aim of the study is to determine the concentration of 8-isoprostane in the blood of rats after implantation of surgical meshes coated with tantalum and tantalum oxide. Materials and methods. The experimental group included 36 male rats of the WAG population weighing 240 ± 20 g. A 15x15 mm polypropylene surgical mesh was surgically implanted between the abdominal wall and various sections of the colon. Anesthesia was performed using the intraperitoneal route of administration of the drug «Relax» BioTestLab, Ukraine) at a dose of 8 mg/kg. After 28 days, the experimental animals were decapitated by cervical dislocation, and blood was immediately collected in sterile K2 EDTA VACUTAINER tubes (BD Vacutainer®). Blood samples were used to determine the content of 8-isoprostane in the blood by enzyme-linked immunosorbent assay using the «8-isoprostane EIA KIT» kit (Cayman Chemical Company, USA) according to the manufacturer’s instructions on the «Solar» PV-1251C spectrophotometer. Results and discussion. The content of 8-isoprostane in blood plasma was statistically 158,6 % higher in the experimental group implanted with uncoated surgical mesh and 171,5 % higher in the experimental group implanted with polypropylene surgical mesh with tantalum nitride coating in comparison with the results of the control group. The concentration of 8-isoprostane in the blood of rats after implantation of polypropylene surgical meshes with a coating based on tantalum and tantalum oxide was higher by 102,6 % and 111,1 %, respectively, compared to the results obtained in the group of intact animals, which indicates their better biocompatibility compared to the results of the groups implanted with uncoated and coated tantalum nitride surgical mesh. Conclusion. The obtained results indicate that coatings based on tantalum and tantalum oxide inhibit lipid peroxidation and demonstrate excellent biocompatibility and anti-inflammatory effect, which makes it possible to consider their future use in hernioplasty.

A Copeptin as a Predictor Marker for Insulin Resistance Among Women with Polycystic Ovary Syndrome

Background: A polycystic ovarian syndrome (PCOS) is a common endocrine syndrome in which women have a wide range of clinical presentations; insulin resistance was linked to its pathogenesis. Objective: We aimed to investigate the copeptin role as a predictive marker of insulin resistance among PCOS women. Material and Methods: In University Hospital, we included 280 women, with 140 of them being healthy controls. 140 out of 280 cases of PCOS subdivided into two groups depending on the insulin resistance; group 1 with homeostasis model assessment for the insulin resistance &lt; 2.5. Group 2 with homeostasis model assessment for the insulin resistance &gt;2.5. The evaluation of body mass index and blood pressure for all besides the blood sampling for estimation of a follicular stimulating hormone, luteinizing hormone, prolactin, estradiol, sex hormone-binding globulin, total testosterone, fasting insulin dehydroepiandrosterone sulfate, C-reactive protein, plasma glucose, free androgen index, and plasma copeptin using the Copeptin-Human EIA Kit besides the transvaginal ultrasound for ovarian assessment. Results: When compared to other groups, PCOS women with positive insulin resistance &gt;2.5 had a significantly higher plasma copeptin level. The ROC curve calculated a 1.94 pmol/L; plasma copeptin cutoff value for detecting the insulin resistance in PCOS with 88 % sensitivity value and 36 % specificity, AUC was 0.88. Conclusion: The significant positive relationship between serum copeptin and insulin resistance with high sensitivity implies its usefulness as a marker of insulin resistance among PCOS patients with a high prediction of its complication.

Salivary vitamin D levels among OSCC and normal Indian patients.

Oral Squamous Cell Carcinoma comprises nearly 90% of all Oral cancers. Recent studies on Oral Squamous Cell Carcinoma are done with a focus on identification of factors that affect the outcome of treatment, one of which is vitamin D levels. The aim of this study was to evaluate vitamin D3 levels in patients with Oral Squamous Cell Carcinoma and Normal Population with saliva as a biomarker. The study sample comprised of 40 subjects, of whom 20 were patients diagnosed with OSCC and 20 healthy subjects. The salivary samples obtained were assessed using Vitamin D3 ELISA kit. The mean salivary Vitamin D3 levels were 39.09 ng/dl among OSCC patients and 44.49 ng/dl among healthy subjects. Salivary Vitamin D levels in OSCC patients were found to be significantly less compared to the healthy controls. More than 90% of cancers of the oral cavity are Oral Squamous Cell Carcinoma (OSCC). Recent studies are done with a focus on identification of factors affecting the treatment of cancer, one of which is vitamin D levels. The aim of the study was to evaluate salivary vitamin D3 levels in patients with Oral Squamous Cell Carcinoma and in Normal Population. 40 patients were selected for the study. Salivary samples collected were assessed using Vitamin D3 ELISA Kit-EDI Total 25-OH Vitamin D EIA Kit to detect the salivary vitamin D levels. Mean Salivary Vitamin D3 levels were found to be 39.09 ng/dl in OSCC patients and 44.49 ng/dl in healthy subjects. A salivary Vitamin D level in OSCC patients was found to be significantly less compared to the healthy controls. Decreased Vitamin D3 levels in patients with OSCC indicate that Vitamin D3 deficiency increases the rate of cell proliferation, invasion, angiogenesis & metastasis in patients with OSCC. Vitamin D level is likely to reduce the severity of the disease and hence Vitamin D supplementation for OSCC patients in the early stages of treatment and after treatment can be done to enhance the prognosis.

Evaluation of the Aspergillus Galactomannan ag VircliaR Monotest test as an alternative to Platelia™ Aspergillus EIA kit

The Aspergillus Galactomannan Ag Virclia® (GMVClia) test is a monotest and automated galactomannan technique based on chemiluminescent immunoassay (CLIA). To evaluate the performance of the GM-VClia test in serum and bronchioalveolar lavage (BAL) samples previously processed with the Platelia ™ Aspergillus EIA kit (GM-Plat). 56 samples of serum 40 from BAL (some of them with galactomaman determination in both samples), from patients with pulmonary diseases, hematological diseases, SLE, Covid-19 and tumors, among others, were studied. Thirteen patients had invasive aspergillosis (1 proven and 12 probable). The correlation between both methods for serum and BAL was r = 0.8861 p < 0.0001 and r = 0.6368 p < 0.001, respectively. There was a global concordance of 67.7% (65/96), being 85.7% (48/56) in sera and 42.5.0% (14/49) in BAL. By raising the cut-off point in LBA by GM-VClia, the agreement increased to 85.7%. A greater correlation and concordance was observed in sera than in BAL. The GM-VClia kit had a higher sensitivity and NPV than the GM-Plat kit. The disadvantages of GM-VClia are the "doubtful" category, which makes interpretation difficult and that with the current cut-off points in LBA the correlation with GM-Plat is lower. The advantages are its greater sensitivity, ease of processing and faster results.

Impact of Indigestible Materials on the Efficiency of Fecal Corticosterone Immunoassay Testing in Pituophis Species.

Simple SummaryEvaluating stress in animals is important for improving animal welfare and husbandry. However, it has been challenging to establish reliable and noninvasive methods for quantifying stress. Steroid hormones released during prolonged periods of stress are metabolized by the liver and excreted in feces. In snakes, corticosterone is the primary hormone of stress and is often measured in fecal samples collected from these animals. Assays can be used to measure the metabolites of corticosterone from feces, but are limited by compounding factors (season, reproductive status, diet, etc.) that influence the ability to confidently conclude the results. Our overall objective is to standardize these methods by first improving the extraction methodology. We found that 75% of the fecal sample contained indigestible materials (hair, teeth, bone, etc.) and, therefore, interferes with the extraction process. After removing the indigestible material, we found that we had a 95% improvement in overall yield. These findings alleviate one limitation to using fecal samples to measure stress in animals. Measuring fecal glucocorticoid metabolites (FGM) has recently become a sought-after method for assessing stress in animals. While there are many benefits to this methodology, there are also recognized limitations, including the apprehensive interpretation of results. While many factors can influence FGM levels, we aimed to standardize and improve these methods in snakes. Fecal samples were collected from Pituophis species and FGMs were extracted by two different sample collection methods: (1) fecal sample containing undigested materials and (2) fecal samples with undigested materials removed. These extracts were then used to quantify FGM concentrations using a corticosterone EIA kit. The results indicated that the samples with the undigestible materials removed had a 95% increase in overall yield (p < 0.01). Since the collected fecal samples contain 75% undigestible materials by weight, these results support our hypothesis that removing these materials will improve extraction methods for a more reliable measurement of corticosterone. This is the first step towards standardizing the methods for assessing stress by measuring fecal glucocorticoid metabolites in snakes.

Effectiveness of immunization activities on measles and rubella immunity among individuals in East Sepik, Papua New Guinea: A cross-sectional study

ObjectivesThis study aimed to assess measles and rubella immunity by measuring virus-specific immunoglobulin G (IgG) prevalence among individuals and evaluate the effectiveness of recent supplementary immunization activities (SIAs) by comparing the antibody positivity rates of the SIA target age groups in 2015 with those in 2019 as measles and rubella are endemic in Papua New Guinea. MethodsA cross-sectional study. The measles- and rubella-specific IgG levels of patients aged ≥1 year at two clinics in East Sepik province, Papua New Guinea were assessed with commercially available virus-specific IgG EIA kits. ResultsIn total, 297 people participated in the study and 278 samples with sufficient volume, relevant information, and age inclusion criteria were analyzed. The overall IgG prevalence rates were 62.6% for measles and 82.0% for rubella. The age groups targeted in the 2019 SIAs had a higher IgG prevalence than those targeted in the 2015 SIAs for both the infectious diseases. Moreover, the IgG prevalence for rubella was higher than measles in these groups. ConclusionsThe anti-measles and anti-rubella IgG prevalence in the target groups were lower than those required for herd immunity. The immunization program should be emphasized to eliminate measles and rubella. Further population-based studies are warranted.

Factors influencing the welfare of dairy goats

A variety of stressful situations commonly occur on dairy farms which can impair the well-being of the animals. The aim of this study was to analyse the concentration of cortisol in the saliva of dairy goats and on the basis thereof to determine the degree of stress experienced by them in relation to selected situations on farms. The following situations were selected as stressful: first visit to the milking parlour; weaning off; loading and transport; deworming; and the disruption of social hierarchy. We examined 344 samples from 100 animals using cotton swabs for the saliva collection. Commercially available ELISA kits (Cortisol EIA Kit, BosterBio, California, USA) which can detect cortisol in the saliva of all animal species, were used for the analysis. During the first visit to the milking parlour, weaning off, deworming and disruption of social hierarchy there was a significant (P&lt; 0.05) increase in cortisol concentrations compared to the basal values. For loading and transport there was a highly significant (P&lt; 0.01) increase in cortisol concentrations compared to the basal values. Although these situations are inevitable on farms, efforts should be made to eliminate them as much as possible due to the stress the animals experience in them.

Fecal Cortisol Dynamics of Captive Giraffes in Zoos in Northern Japan

Animals in zoos are kept under environmental conditions differing from those of their original habitat. Recently, zoos have been attempting to reduce stress by introducing environmental enrichment. Adrenal-derived glucocorticoids are common stress indicators found in blood, saliva, urine, and feces. Fecal cortisol levels have been studied as a stress marker in captive giraffes, but the data are insufficient. In this study, we examined fecal cortisol in captive giraffes in northern Japan to increase basic data on fecal cortisol as an indicator of environmental enrichment. For one month, we collected fecal samples from 14 giraffes [Masai giraffes (Giraffa camelopardalis tippelskirchi) and reticulated giraffes (Giraffa camelopardalis reticulata)] from eight zoos. The fecal samples were stored at –20 °C until fecal cortisol levels were measured using a cortisol EIA kit after drying, grinding, and ethanol extraction. Cortisol levels were compared between males and females, pregnant and nonpregnant animals, by age, and by differences in temperature of the rearing environment. The fecal cortisol levels ranged from 37~10346 pg/g. Among females, pregnant individuals had the highest values. Among nonpregnant females, a female that was harassed by a male had higher fecal cortisol levels than the others. This may be due to chronic psychological distress. In males, values varied greatly among individuals. This study increases the volume of giraffe fecal cortisol data, which will contribute to establishing basic giraffe fecal cortisol values.

Sampling methods for determination of cortisol in pig saliva and their use in the assessment of pig welfare

This study focused on finding new information regarding the assessment of pig saliva cortisol samples in terms of practical effects of the sampling, sample storage conditions, and their laboratory analysis. The study was divided into two experiments. The first experiment was focused on finding the effect of sampling time on cortisol concentrations in pig saliva. The second experiment was focused on determining the effect of storage conditions on the value of salivary cortisol. Before the initiation of the study, we tested which one of the commercially available ELISA kits would be the most suitable for our experiments. Simultaneously, we carried out a pre-study to evaluate the effect of relocation and change in the housing type on the concentration of salivary cortisol in gestating sows. The samples were obtained by oral cavity swabbing, using a standard cotton swab. In the first study, piglets were examined at the age of 4 ± 1 days, and breeding management routine procedures were used as a stress factor. In the second study, the piglets were examined immediately after weaning (at 28 ± 2 days of age). The Cortisol EIA kit was found to be statistically more accurate and thus a more suitable ELISA kit for our experiment. Analysis of the relocation effect and the effect of change in the housing type showed that relocation does not seem to be a stress factor for gestating sows as no significant changes were observed in salivary cortisol concentration (P &gt; 0.5); however, the change in the housing type lead to a significant increase in salivary cortisol (P &lt; 0.001). In the first study, we determined using the ELISA method that the most significant difference occurred in 40 min (P &lt; 0.01), which suggests that the best time for a sampling in order to assess salivary cortisol concentration is 40 min after stress induction by routine procedures. The conclusion of the second study was that in the monitored period of 60 h (P &lt; 0.05), cortisol concentration decreased depending on the storage temperature. The decrease started between 48 and 60 h which showed that cortisol is stable in the saliva sample for at least 48 h. These findings will be further applied in our following studies focused on assessment of salivary cortisol concentration after stress induction.

Association of leukotrienes and prostaglandins with splenic 18F-FDG uptake in hepatobiliary cancer patients.

In this study, we aimed to evaluate the relationship between lipid metabolites and diffuse splenic 18F-FDG uptake with the means of leukotriene (LT) and prostaglandin (PG). We enrolled 36 patients with hepatobiliary malignancies who underwent fluorine-18-fluorodeoxyuglucose (18F-FDG) positron emission tomography/computed tomography (PET/CT) for staging workup. Patients were divided into two groups according to spleen to liver ratio (S/L ratio) of 18F-FDG uptake. Blood sample of each patient was collected on the day of conducting PET/CT scanning. Several types of LT and PG, including LTB4, LTC4, LTE4, PGD2, PGE2, and PGF2α were measured from blood plasma samples from 36 patients using enzyme immunoassay (EIA kit, Cayman Chemical Co.). White blood cell counts (P=0.0176) and C-reactive protein (P=0.0036) were higher in patients with splenic 18F-FDG uptake exceeding hepatic 18F-FDG uptake. Among the several types of PG and LT, PGD2 (P=0.0033) was higher in patients with hepatic 18F-FDG uptake exceeding splenic 18F-FDG uptake, however, LTC4 (P=0.0237) and LTE4 (P=0.0429) were higher in patients with splenic 18F-FDG uptake over hepatic uptake. Higher levels of LTC4 and lower levels of PGD2 are shown in patients with splenic 18F-FDG uptake. If the clinician incidentally finds splenic 18F-FDG uptake exceeding hepatic uptake, concurrent inflammation should be considered.

Individualized approach to predicting complications of acute myocardial infarction in patients with chronic obstructive pulmonary disease

Objective. Development of an algorithm for predicting the complicated course of acute MI (rhythm disturbances, acute left ventricular failure: pulmonary edema, cardiogenic shock) in patients with COPD.Materials and methods. 37 patients with acute myocardial infarction on the background of COPD were examined, undergoing inpatient treatment in the conditions of the regional vascular center of the State Budgetary Healthcare Institution of the Alexandro-Mariinsky Regional Clinical Hospital in 2017–2019. Clinical examination included assessment of complaints, life history and illness. The enzyme-linked immunosorbent assay was used to determine: the concentration of the HSP 70 protein using the HSP 70 High Sensitivity EIA Kits (Stressgen, USA), interleukins IL‑1β, IL‑2, IL‑6 with reagent kits of VEKTOR-BEST JSC (St. Novosibirsk, Russia) and neopterin test with the Neopterin ELISA kit (IBL International, Germany). Determination of the content of apoptotic cells from heparinized venous blood was performed using the Annexin-V-FITC / 7AAD reagent kit (Beckman Coulter, USA). Statistical data processing was carried out using the SPSS 26.0 (USA).Results. When analyzing the frequency of occurrence of the studied laboratory diagnostic signs, significant differences were found for the indices of circulating annexin V mononuclear cells at an early stage of apoptosis and neopterin. Based on the data obtained and the selection of predictors, the probability of complications (rhythm disturbances, acute left ventricular failure) was calculated using the logistic regression equation. Using ROC analysis, a cut-off was determined for the levels of circulating annexin V mononuclear cells at an early stage of apoptosis and neopterin.Conclusion. Information on the estimated high risk of developing complications of acute myocardial infarction, such as rhythm disturbances and acute left ventricular failure, will help to purposefully select the amount of preventive and therapeutic measures in patients with acute MI associated with COPD to minimize this risk.

Study on Efficacy Evaluation of Deramal Bioactive Properties of the Prunus tomentosa Fruits Fermented with Lactobacillus acidphilus

The materials fermented with natural products have been used in a variety of cosmetic applications as a whitening, wrinkle and inflammatory agent. &lt;i&gt;Prunus tomentosa&lt;/i&gt; fruits have been used as a traditional medicine for anti-oxidant and have been reported to possess anti-microbial properties. This study investigated the deramal bioactive properties of the &lt;i&gt;Prunus tomentosa&lt;/i&gt; fruits fermented with &lt;i&gt;Lactobacillus acidphilus&lt;/i&gt; (PTF). The antioxidant activity of the PTF was measured by 1,1-diphenyl-2-picryl hydrazyl (DPPH) method. The cytotoxicity of human dermal fibroblast cell, murine macrophage cell line RAW 264.7 and B16-F10 melanoma cells were evaluated by sulforhodamine B (SRB) assay. Inhibitory effect of the PTF on nitric oxide (NO) prodction was evaluated by Griess Reagent System. The collagen synthesis was evaluated by using procollagen type-IC peptide (PIP) EIA kit. The tyrosinase activity, inhibitory effect of elastase and tumor necrosis factor-α (TNF-α) induced matrixmelloproteinase-1 (MMP-1) were also evaluated. It found that the PTF showed antioxidant activity without showing significant cytotoxicity in tested cells. The PTF were downregulated NO production in LPS-stimulated macrophages with nocytotoxicity (p&lt;0.05). The PTF significantly induced collagen synthesis from the skin fibroblasts (p&lt;0.05) and also dramatically inhibited MMP-1 activity (p&lt;0.05). The PTF were inhibited activity of elastase and dramatically decreased melanin synthesis with inhibition of tyrosinase activity (p&lt;0.05) and didn’t show cytotoxicity in B16-F10 melanoma cells. Therefore, the PTF showed bioactive properties on anti-oxidant, whitening, skin elasticity and anti-wrinkle efficacy. The PTF may be an appropriate choice for cosmetic products that aim to provide dermal bioactive effects.

Vitamin D level, lipid profile, and vitamin D receptor and transporter gene variants in sickle cell disease patients from Kurdistan of Iraq.

IntroductionSickle cell disease (SCD) patients are susceptible to the development of vitamin D deficiency (VDD). Vitamin D through binding to vitamin D receptor (VDR) exerts its function and affects gene transcription in target tissues. VDR gene variants affect bone mineral density.MethodsIn a case‐control study, 101 SCD patients including 61 sickle cell anemia (SCA), 39 S/β‐thalassemia, and 1 HbS/HbD (SD) along with 110 healthy individuals from Kurdistan of Iraq were studied. The lipid profile, vitamin D level, FokI, and TaqI variants of VDR and group‐specific component (GC) were detected using the standard enzymatic method, the immunodiagnostic systems limited EIA kit and PCR‐RFLP methods, respectively.ResultsAround 93% and 82% of SCA and S/β‐thalassemia patients, respectively, had VDD compared to 83% of healthy individuals. Severe VDD (<10 ng/ml) was detected in 78.7% of patients with HbSS. Plasma levels of total cholesterol, HDL‐C, and LDL‐C in SCD patients were significantly lower compared to controls. Vitamin D levels were negatively correlated to TG and positively correlated to total cholesterol and HDL‐C. The frequencies of the C allele of FokI were 81.7% (p = 0.003), 80.3% (p = 0.034), and 84.6% (p = 0.011) in all SCD, SCA, and S/β‐thalassemia patients, respectively, compared to 69.1% in controls. However, no significant difference was detected comparing the frequencies of VDR TaqI and GC polymorphisms between SCD patients and controls.ConclusionIn the present study, we found hypocholesterolemia, high prevalence of VDR FokI C allele, and low vitamin D levels among children and adults with SCD from Kurdistan of Iraq.

The seroprevalence of rubella virus in children between 0-15 years old in a hospital in Ankara province before expanded immunization program

Aim: In this study, it was aimed to determine the seroprevalence of rubella in children aged 0-15, who was admitted to Ankara Training and Research Hospital Pediatric outpatient clinic for various reasons between December 2001 and July 2002. In this descriptive study, it is thought that knowing the seroprevalence before the national vaccination program will be important for the evaluation of the future data. Material and Method: In this descriptive study; the seroprevalence of rubella in 1170 children aged 0-15 years was investigated. The age, gender, socio-economic level and number of siblings of the 954 cases included in the study were recorded on the prepared forms, and the history of infection was taken from their families. Rubella specific IgG levels from serum samples of the cases were measured by using the RADIM rubella IgG EIA kit with Micro-ELISA technique. Results: The average age of the children included in the study was found to be 76 } 49 months. Considering their gender, 56.9% (n=543) were male and 43.1% (n=411) were female. Considering their socio-economic levels; 550 children were in the low-income group; 393 children were in the middle-income group and 11 children were in the high-income group. When the distributions are evaluated according to the number of siblings; 14% (n=134) of the cases were a single child, 45% (n=426) had one sibling, 28% (n=267) had two siblings, and 13% (n=127) of the cases had ≥ 3 siblings. When evaluated in terms of rubella seroprevalence, 47.2% (n=450) of the cases had negative rubella IgG and 52.8% (n=504) of them had positive rubella IgG. According to the age the seropositivity showed a significant increase with the increasing age and remained constant at around 91% in the 13-15 age range (p &lt;0.05). Conclusion: This study, conducted before the National Immunization Program in 2003, showed that a large part of our population is vulnerable to the risk of rubella infection. Considering that the seroprevalence increases with age in order to prevent rubella infection, which affects women of reproductive age and congenital rubella syndrome (CRS) seen in newborns in particular, it is important to vaccinate all children who have completed their 12th month and have entered the 48th month within the scope of the national vaccination program. The data of this study will guide us in evaluating the success of the program in the population, we provide healthcare services in the following years, as it reflects the situation before the Expanded Immunization Program (EIP).