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Centaurea thracica (Janka) Hayek is a plant common in southern Bulgaria. The inflorescences were collected during June and September 2021, while their seeds were obtained in September 2021. The chemical and lipid composition of the inflorescences during the vegetation process of the plant were established. A significant decrease in total proteins (from 8.7 to 7.4%), glyceride oils (2.0-1.7%), and ash (4.5-4.2%) content was observed, while the amount of carbohydrates (72.3-77.2%) and fibers (28.7-35.8%) increased. During the vegetation of the plant, the content of oleic and linoleic acids increased up to 2-3 times, while the level of palmitic acid decreased. The lipids from the seeds were rich in oleic (53.0%) and palmitic (36.2%) acids. The tocopherol content in the oils of the inflorescences during vegetation increased from 58 to 110 mg/kg, and the content in the oil from the seeds was 260 mg/kg. The phospholipid content decreased during vegetation, and differences were observed in the composition between the inflorescences and the seeds. The high content of oleic acid, linoleic acid, tocopherols, and phospholipids determine the nutritional and biological value of the oils isolated from Centaurea thracica , and contribute to their potential use in various directions.

Chemerin and resistin are adipokines studied as potential markers for early diagnosis and disease severity in patients with knee osteoarthritis (KOA) Therefore, we aimed to investigate the associations serum and synovial levels of chemerin and resistin with inflammatory parameters and ultrasonographic scores (US) in KOA individuals. Serum was collected from 28 patients with KOA and synovial fluid was obtained from 16 of them. Another 31 age and sex matched cases with no joint disease were included as healthy controls. Concentrations of chemerin, resistin, interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) were determined with ELISA. Erythrocyte sedimentation rate (ESR), C-reactive protein, serum uric acid (UA) were measured in the patients group. Participants with KOA underwent US assessment using the Outcome Measures in Rheumatology (OMERACT) scores. Patients with KOA had statistically significant higher level of serum resistin than healthy controls [11.05 (3.78-24.13) ng/mL and 7.23 (3.83-12.19) respectively, p < 0.001]. A strong correlation was found between serum chemerin and ESR (r = 0.434, p = 0.021), uric acid (r = 0.573, p = 0.001) as well as the US (r=-0.872, p < 0.001). Serum resistin demonstrated significant association with TNF-alpha (r = 0.398, p = 0.044). In conclusion, both chemerin and resistin might contribute to inflammatory changes associated with KOA. Further studies are needed to elucidate their potential role in the pathogenesis of the disease., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Present scientific evidences about the biological activity and potential medical application of extracts derived from Marrubium friwaldskyanum Boiss. are limited. Therefore, our study was undertaken to define several main characteristics in this regard - in vitro cytotoxicity and antitumor properties, antibacterial activity and immunomodulatory potential. Extracts were obtained from different aerial parts of Marrubium friwaldskyanum - stems, leaves and flowers. The in vitro cytotoxicity and antitumor activity of the samples were evaluated by tetrazolium salt reduction tests and Neutral red uptake assays using four human cell lines (a normal fibroblastic and three adenocarcinoma cell lines/A549, HeLa, HT-29/) and by experiments with HT-29 tumor spheroids. Antibacterial activity toward Gram-negative ( Escherichia coli ) and Gram-positive ( Bacillus cereus ) species was assessed based on estimation of minimal inhibitory and minimal bactericidal concentrations as well as longitudinal studies on bacterial viability. Ex vivo assays with normal leukocytes were performed to define potential immunomodulatory activity of the extracts. Our results demonstrated selective antitumor activity of the extracts directed against colon adenocarcinoma HT-29 cells and cervical adenocarcinoma HeLa cell line. Metabolic activity of A549 lung adenocarcinoma cells was affected only by the sample derived from flowers. M. friwaldskyanum leaf and flower extracts showed the highest activity, which included reduction of HT-29 tumor spheroid growth and viability. The studied samples exhibited antibacterial activity against both bacterial species tested. Treatment with M. friwaldskyanum extracts affected specific leukocyte populations (HLA + , CD19 + , CD11b + , CD25 + cells). These results demonstrate for the first time complex biological effects of extracts derived from M. friwaldskyanum and their potential to serve as a source of valuable compounds for the pharmaceutical industry., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors. Published by Elsevier Ltd.)

Extracts from medicinal plants are widely used in the treatment and prevention of different diseases. Micromeria frivaldszkyana is a Balkan endemic species with reported antioxidant and antimicrobial characteristics; however, its phytochemical composition is not well defined. Here, we examined the metabolome of M. frivaldszkyana by chromatography-mass spectrometry (GC-MS), ultra-performance liquid chromatography-mass spectrometry (UPLC-MS-MS), and inductively coupled plasma mass spectrometry (ICP-MS). Amino acids, organic acids, sugars, and sugar alcohols were the primary metabolites with the highest levels in the plant extract. Detailed analysis of the sugar content identified high levels of sucrose, glucose, mannose, and fructose. Lipids are primary plant metabolites, and the analysis revealed triacylglycerols as the most abundant lipid group. Potassium (K), magnesium (Mg), zinc (Zn), and calcium (Ca) were the elements with the highest content. The results showed linarin, 3-caffeoil-quinic acid, and rosmarinic acid, as well as a number of polyphenols, as the most abundant secondary metabolites. Among the flavonoids and polyphenols with a high presence were eupatorin, kaempferol, and apigenin-compounds widely known for their bioactive properties. Further, the acute toxicity and potential anti-inflammatory activity of the methanolic extract were evaluated in Wistar rats. No toxic effects were registered after a single oral application of the extract in doses of between 200 and 5000 mg/kg bw. A fourteen-day pre-treatment with methanolic extract of M. frivaldszkyana in doses of 250, 400, and 500 mg/kg bw induced anti-inflammatory activity in the 1st, 2nd, and 3rd hours after carrageenan injection in a model of rat paw edema. This effect was also present in the 4th hour only in the group treated with a dose of 500 mg/kg. In conclusion, M. frivaldszkyana extract is particularly rich in linarin, rosmarinic acid, and flavonoids (eupatorin, kaempferol, and apigenin). Its methanolic extract induced no toxicity in male Wistar rats after oral application in doses of up to 5000 mg/kg bw. Additionally, treatment with the methanolic extract for 14 days revealed anti-inflammatory potential in a model of rat paw edema on the 1st, 2nd, and 3rd hours after the carrageenan injection. These results show the anti-inflammatory potential of the plant, which might be considered for further exploration and eventual application as a phytotherapeutic agent.

The prolonged consumption of a high-fat diet (HFD) leads to abnormal growth of the visceral adipose tissue (VAT), increased macrophage infiltration, and altered secretion of biologically active molecules. This is considered as a precondition for the development of obesity, inflammation, and obesity-related disorders. Therefore, we studied HFD-induced changes in the tissue levels of the inflammatory markers C-reactive protein, serum amyloid-A, and interleukin-4 in healthy male Wistar rats. The animals were first divided at random into two groups subjected to either a standard or a high-fat diet. The initial effect of the diet was evaluated after fourteen weeks. In order to study the diet duration effect, the standard diet was given to twelve animals from the HFD group, while the remaining continued with the HFD for an additional four weeks. Our results showed that the HFD barely affected body mass index, conicity, relative fat mass, and Lee indices, whereas it provoked adipocyte hypertrophy and gradually increased the levels of both the pro- and anti-inflammatory markers. The switch from the high-fat to the standard diet resulted in the comparatively fast restoration of the baseline levels of the studied molecules. Although, the prolonged consumption of an HFD causes adipocyte hypertrophy in healthy male animals, the inflammatory process in VAT is well-coordinated, time-dependent, and reversible.

Objective: Trace elements are essential for the biochemistry of the cell. Their reference values have been found to differ considerably in pregnant women stratified by age, place of residence, anthropometric status, and length of pregnancy. In optimal amounts, these elements reduce the risk of pregnancy complications. Subclinical hypothyroidism in pregnancy is associated with adverse maternal and neonatal outcomes. The aim of the study was to determine the effects of zinc (Zn), copper (Cu), magnesium (Mg), and rubidium (Rb) on pregnant women in an iodine deficiency region and find the relationship with the thyroid status and nutrition., Methods: We evaluated the iodine status of 61 healthy pregnant women from an iodine deficient region in Bulgaria. Thyroid stimulating hormone (TSH) and thyroxin free (FT4) levels were measured using ELISA., Results: We found elevated levels of copper that differed the most between the first and second trimesters; Cu and TSH were found to be positively correlated (р < 0.05). Lower Cu levels were found in pregnant women consuming pulses more than 2-3 times a week (р = 0.033). The women consuming fish more than 2-3 times a week had higher levels of Rb. We found a pronounced iodine deficiency in more than half of the examined women in the first to third trimesters, without any effect of pregnancy on the ioduria (р=0.834). All second and third trimester cases were associated with severe ioduria (< 150 µg/L)., Conclusion: The high Cu levels were associated with subclinical hypothyroidism (SCH) and less pulse consumption during pregnancy in an iodine deficiency endemic area. SCH was found in 24% of the pregnant women in such an area while in 13% of them SCH had progressed to overt hypothyroidism.

Marrubium species have been used since ancient times as food additives and curative treatments. Their phytochemical composition and various pharmacological activities were the focus of a number of scientific investigations but no comprehensive metabolome profiling to identify the numerous primary and secondary metabolites has been performed so far. This study aimed to generate a comprehensive picture of the total metabolite content of two Marrubium species- M. peregrinum and M. friwaldskyanum -to provide detailed information about the main primary and secondary metabolites. In addition, the elemental composition was also evaluated. For this purpose, non-targeted metabolomic analyses were conducted using GC-MS, UPLC-MS/MS and ICP-MS approaches. Nearly 500 compounds and 12 elements were detected and described. The results showed a strong presence of phenolic acids, flavonoids and their glucosides, which are generally of great interest due to their various pharmacological activities. Furthermore, tissue-specific analyses for M. friwaldskyanum stem, leaves and flowers were carried out in order to outline the sources of potentially important bioactive molecules. The results generated from this study depict the Marrubium metabolome and reveal its dual scientific importance-from one side, providing information about the metabolites that is fundamental and vital for the survival of these species, and from the other side, defining the large diversity of secondary substances that are a potential source of phytotherapeutic agents.

Introduction : Numerous studies to date have shown that the development of dysbiotic gut microbiota is a characteristic finding in chronic kidney disease (CKD). A number of uremic toxins progressively accumulate in the course of CKD, some of them generated by the intestinal microbiome, such as indoxyl sulfate (IS) and p-cresyl sulfate (p-CS). They are found to be involved in the pathogenesis of certain complications of uremic syndrome, including low-grade chronic inflammation and oxidative stress. The aim of the present study is to research the serum concentration of IS and p-CS in end stage renal disease (ESRD) patients undergoing conventional hemodialysis, as well as to study the possibilities of influencing some markers of inflammation and oxidative stress after taking a synbiotic. Materials and Methods : Thirty patients with end-stage renal disease (ESRD) undergoing hemodialysis treatment who were taking a synbiotic in the form of Lactobacillus acidophilus La-14 2 × 10 11 (CFU)/g and prebiotic fructooligosaccharides were included in the study. Serum levels of total IS, total p-CS, Interleukin-6 (IL-6), and Malondialdehyde (MDA) were measured at baseline and after 8 weeks. Results . The baseline values of the four investigated indicators in the patients were significantly higher-p-CS (29.26 ± 58.32 pg/mL), IS (212.89 ± 208.59 ng/mL), IL-6 (13.84 ± 2.02 pg/mL), and MDA (1430.33 ± 583.42 pg/mL), compared to the results obtained after 8 weeks of intake, as we found a significant decrease in the parameters compared to the baseline-p-CS (6.40 ± 0.79 pg/mL, p = 0.041), IS (47.08 ± 3.24 ng/mL, p < 0.001), IL-6 (9.14 ± 1.67 pg/mL, p < 0.001), and MDA (1003.47 ± 518.37 pg/mL, p < 0.001). Conclusions : The current study found that the restoration of the intestinal microbiota in patients with CKD significantly decreases the level of certain uremic toxins. It is likely that this favorably affects certain aspects of CKD, such as persistent low-grade inflammation and oxidative stress.

Objectives: Roma population is one of the major ethnic groups in the Central and Eastern Europe, having high rates of chronic diseases and associated risk factors related to their poor social conditions, unhealthy lifestyle and low educational level. The purpose of our study was to assess the health status of Roma from South Bulgaria by means of blood indicators and determine the prevalence of some cardiovascular (CV) risk factors in the Roma population sample., Methods: The study group consisted of 60 Roma (23 men and 37 women), mean age 53.7 ± 15.9 years, and the control group consisted of 68 non-Roma from the majority population (29 men and 38 women), mean age 45.8 ± 12.2 years. The data were collected via questionnaire, anthropometric measures, and venous blood samples analyses after an overnight fasting., Results: The Roma population subjects were slightly but significantly older compared to the non-Roma group and both study groups included more women. The fasting glucose, body mass index (BMI), triglycerides (TG), total cholesterol (TC), and LDL-cholesterol (LDL-C) levels were significantly higher, and HDL-cholesterol (HDL-C) levels were significantly lower in Roma compared to the control non-Roma group. The values of cardiovascular risk markers such as TC/HDL-C and TG/HDL-C ratios, atherogenic index of plasma (AIP) and lipoprotein combine index (LCI) were significantly higher in Roma compared to non-Roma subjects. The prevalence of obesity in Roma was 35%, diabetes mellitus was recorded in 16.7% of the entire Roma sample, and hyperglycaemia in non-diabetics was recorded in 32%. Hypercholesterolaemia was found in 90% and hypertriglyceridaemia was found in 88.3%. The prevalence of cardiovascular diseases (CVD) was high and was encountered in 71.7% of the Roma participants and most of the subjects (96.7%) reported family history of CVD. The studied population showed high smoking rates with 33.3% active smokers., Conclusions: Our study confirmed high prevalence of CV risk factors among Roma population, such as abnormal lipid profile parameters, obesity and heavy smoking and very high cardiovascular morbidity rate. Therefore, adequate measures and healthcare programmes aiming at the early identification, treatment and prevention of CVD risks among Roma are necessary.

Matrix metalloproteinases (MMPs) are a large family of Ca 2+ and Zn 2+ dependent proteolytic enzymes, able to cleave the various components of the extracellular matrix (ECM), as well as a range of other regulatory molecules. Several reports have proven the important role of both MMPs and their endogenous inhibitors, TIPMs, in oral health, the initial development of the tooth, and during enamel maturation. In this mini-review, we aim to summarize the literature information about the functions of MMPs, paying more attention to MMP-8 (collagenase-2 or neutrophil collagenase) in the development and progression of periodontitis, peri-implantitis, and carious lesions. We also emphasize the role of particular gene variants in MMP8 as predisposing factors for some oral diseases.

Introduction: Stress is a phenomenon accompanying everyday life. Various reactions of the body are activated during stress, including activation of the hypothalamo-pituitary-adrenal endocrine axis and the autonomous nervous system. Their activity could be assessed by the measurement of biological stress marker molecules - salivary α-amylase and cortisol - on one side and heart rate, and blood pressure monitoring, on the other. The aim of the present study is to determine whether the daily professional routines cause stress development among general practitioners (GPs) who work in rural, distant areas in the territory of the Republic of Bulgaria., Methods: One hundred and twenty-eight GPs were asked to answer a questionnaire to assess their health conditions, habits, priority objectives and interests, as well as their moral and ethical relationships with the patients. On this basis 40 participants were selected and outlined for the present survey (n=40, mean age 55.92±8.8). Four salivary samples were collected from each participant during 1 week: two on Monday (one in the morning and one after work) and two on Friday, by the same mechanism. The salivary samples collection was followed by measurement of blood pressure and pulse. This non-invasive survey was based on an ELISA method for quantitative determination of the marker molecules in saliva. The blood pressure and pulse were measured by blood pressure monitor., Results: The levels of the salivary α-amylase were significantly higher at the end of the working day, especially on Friday (142.28±23.34 U/mL, p=0.018), but not between the beginning and the end of the week. The normal cortisol awakening response, characterized by a peak in the levels of cortisol after wake-up, followed by a slow decrease during the day, was detected only at the beginning of the week. The data show a significant impairment of this regularity at the end of the week. Moreover, the levels of cortisol, measured for the morning samples, show a decrease at the end of the week (25.73±10.51 ng/mL) in comparison to the beginning of the week (30.1±10.84, p=0.033). The analyses on the effect of smoking (p=0.002) and alcohol consumption (p=0.036) on stress development show a significant increase in the levels of the salivary α-amylase, but not on the levels of salivary cortisol. The changes in the blood pressure indicate stress development at the end of the week (p=0.04), while the pulse showed changes within a day rather than during the week. The values of the pulse were higher at the end of the day., Conclusion: The professional lives of the GPs who work in distant and rural places are associated with stress development. Different habits from the daily routine, such as alcohol consumption, smoking and physical activity, could be considered as modulators of stress development.

Type 1 diabetes mellitus is characterized with decreased microbial diversity. Gut microbiota is essential for the normal physiological functioning of many organs, especially the brain. Prebiotics are selectively fermentable oligosaccharides [xylooligosaccharides (XOS), galactooligosaccharides, etc.] that promote the growth and activity of gut microbes and influence the gut-brain axis. Aerobic exercise is a non-pharmacological approach for the control of diabetes and could improve cognitive functions. The potential beneficial effect of XOS and/or aerobic training on cognition, the lipid profile and oxidative stress markers of experimental rats were evaluated in this study. Male Wistar rats were randomly divided into three streptozotocin-induced diabetic groups and a control group. Some of the rats, either on a XOS treatment or a standard diet, underwent aerobic training. The results showed that the aerobic training independently lowered the total cholesterol levels compared to the sedentary diabetic rats ( p = 0.032), while XOS lowers the malondialdehyde levels in the trained diabetic rats ( p = 0.034). What is more the exercise, independently or in combination with XOS beneficially affected all parameters of the behavioral tests. We conclude that aerobic exercises alone or in a combination with the prebiotic XOS could ameliorate the dyslipidemia, oxidative stress, and cognitive abilities in experimental type 1 diabetic animals., Competing Interests: Conflict of interest: The authors state no conflict of interest., (© 2022 Mariya Choneva et al., published by De Gruyter.)

Background: Extracts obtained from different Betonica species have been shown to possess important biological properties. The present study aimed to investigate the cytotoxicity, antitumor and immunomodulatory potential of the endemic plant Betonica bulgarica (Lamiaceae) and thus, reveal new aspects of its biological activity. Methods: Methanolic extract obtained from inflorescences was analyzed for cytotoxicity against mammalian cell lines. The antitumor potential of the sample was determined using human cervical and lung adenocarcinoma cells (HeLa and A549). Programmed cell death-inducing effects against HeLa cells and peripheral blood lymphocytes, as well as immunomodulatory properties of the extract were determined by flow cytometry analysis. Results: The research results demonstrated that the extract has significant inhibitory potential against HeLa cells (mean IC 50 value 119.2 μg/mL). The sample selectively induced apoptotic death in tumor cells. Cytotoxic effects towards mouse cell lines were detected following treatment with high concentrations of Betonica bulgarica extract (200 and 250 μg/mL). Twenty-four-hour ex vivo incubation of peripheral blood leucocytes in growth medium containing plant extract induced prominent effects in distinct immune cell populations. They included elevated levels of CD25 + and CD56 + T cells' lymphocytes, particularly CD4 + CD25 + and CD8 + CD56 + cells. Conclusions: The present study demonstrates that Betonica bulgarica inflorescence extract possesses potential beneficial antitumor and immunomodulatory activity and could serve as a source of bioactive compounds with biomedical application.

Prebiotics, gut microbiota-fermentable substances, delay the development of type I diabetes. In the present study, we investigated the effect of two prebiotics (galacto-oligosaccharides and xylo-oligosaccharides) on the antioxidant protection, lipid profile, and inflammatory activity of rats with streptozotocin-induced diabetes. The following markers were studied - malondialdehyde, 8-hydroxy-2'-deoxyguanosine, ferric reducing ability of plasma (FRAP), triacylglycerols, total cholesterol (TC), high-density lipoproteins, C-reactive protein (CRP), and interleukin-6. Diabetes was induced in male Wistar experimental rats by streptozotocin injection, while the non-diabetic controls were injected with saline. Afterward the oligosaccharides were administered orally to the experimental animals. The blood collected following the decapitation was analyzed by ELISA. A modified protocol was used only for measuring the FRAP values. The galacto-oligosaccharides and xylo-oligosaccharides lowered the malondialdehyde levels in the diabetic rats ( p  < 0.05). The galacto-oligosaccharides decreased the serum levels of 8-hydroxy-2'-deoxyguanosine ( p  = 0.01), while the xylo-oligosaccharides increased the FRAP ( p  < 0.05) in the experimental animals. None of the oligosaccharides affected triacylglycerol and interleukin-6 concentrations, but the galacto-oligosaccharides decreased the TC and CRP levels in the diabetic animals. Both oligosaccharides exert a beneficial effect on the antioxidant protection of the diabetic rats, but have a minor effect on their lipid and inflammatory profiles., (© 2022 Krasimir O. Boyanov et al., published by De Gruyter, Berlin/Boston.)

The current study investigates the content of sugars, organic acids, phenolic acids and flavonoids, as well as antioxidant and antimicrobial activity of Balkan-endemic Micromeria frivaldszkyana . Glucose was the most abundant sugar in the plant (2.77%), followed by fructose (1.18%) and galactose (0.82%). Eight organic acids were detected with quinic acid being in the highest content-556.3 mg/100 g DW. From the individual phenolic acids, rosmarinic acid was found in the most significant amounts (2040.1 ± 1.97 mg/100 g) and hesperidin was the major representative of flavonoids with content 131.2 ± 5.6 mg/100 g DW. The antioxidant activity of the plant was studied by six methods: 2,2'-diphenylpicrylhydrazyl (DPPH)-286.4 ± 10.43 mM TE/g, 2,2'azinobis (3)-ethylbenzthiazoline-6-sulfonic acid (ABTS)-358.4 ± 10.4 mM TE/g, ferric reducing antioxidant power (FRAP)-388.0 ± 32.4 mM TE/g, cupric reducing antioxidant capacity (CUPRAC)-905.6 ± 19.2 mM TE/g, Oxygen Radical Absorbance Capacity (ORAC)-3250.5 ± 208.1 µmol TE/g and Hydroxyl Radical Averting Capacity (HORAC)-306.1 ± 23.5 µmol GAE/g. In vitro antimicrobial activity against nine microorganism was evaluated but the extract displayed antimicrobial activity only against Listeria monocytogenes ATCC 19111 with inhibition zone diameter 9 mm and minimal inhibitory concentration (MIC) 10 mg/mL.

Introduction: Obesity is linked to the development of low-grade, chronic inflammation. Obesity-related inflammation appears to be a different type of inflammation, mainly due to excessive food intake and unusual homeostasis. It can be evaluated by measuring the concentration of pro- and anti-inflammatory marker molecules – C-reactive protein (CRP), serum amyloid-A (SAA) and interleukin-4., Aim: The aim of the present study is to evaluate the rate of the inflammatory process in heart, provoked by the consumption of a high-fat diet., Materials and Methods: Sixty 8-week-old male Wistar rats were used in this experiment. The laboratory animals were fed orally with two different types of rodent food for 14 or 18 weeks – a high-fat diet (experimental groups) and standard rodent food (control groups). They all were kept under standard housing conditions. The levels of the pro- and anti-inflammatory markers in tissue homogenates from heart were analyzed using ELISA. Their expression in tissue samples was detected immunohistochemically by the biotin-streptavidin-peroxidase method. The total protein concentration was determined by the Lawry method., Results: CRP levels showed no significant differences when the control group was compared with the groups fed with a high-fat diet (p>0.05). The SAA levels detected were also insignificantly changed. Only the IL-4 tissue levels showed tendency to increase (p<0.05) in the high-fat diet group., Conclusions: Our experiment indicates that there is a specific reaction of the heart to a high-fat diet. It also refers to the existence of adaptive mechanisms allowing the heart to counteract the development of dietary induced inflammation., (This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Aim: To find correlations between high thiocyanate and nitrate levels and low selenium levels and the indicators of the iodine and iron status of postpartum women., Materials and Methods: The study included 41 mothers aged 26.4±5.9 yrs from Asenovgrad and nearby villages. Urinary iodine was determined by the Sandell-Kolthoff reaction and thiocyanate - by the interaction of these ions with acidic solution of KMnO4; for serum nitrates we used the colorimetric method; serum selenium was assessed by electro-thermal atomic-absorption spectrophotometry; thyroxin (FT4), the thyroid stimulating hormone (TSH), serum ferritin (SF), and serum transferrin receptor (sTfR) were determined using ELISA; Hb levels were determined by hematology analyzer., Results: Assessing the iodine status, we found a negative correlation between the levels of iodine and thiocyanates in urine (R=-0.717, р<0.0001), a positive correlation between nitrates and TSH (R=0.487, р=0.003) and a negative correlation between nitrates and FT4 (R=-0.312, р=0.06). For the iron status, we found a negative correlation between nitrates and SF (R=-0.429, р=0.009) and between nitrates and Hb (R=-0.383, р=0.021). The Mann-Whitney U-test showed that in women with nitrate levels higher than the mean value there was low FT4 level (р=0.06), high TSH level (р=0.013), low Hb concentration (р=0.061) and low SF concentration (р=0.005). The combined effects of environmental factors (elevated nitrate levels and low selenium level) on the iodine and iron status are manifested by low concentrations of FT4 (р=0.033), Hb (р=0.06) and SF (р=0.05) and high level of TSH (р=0.05). In conclusion, we found that environmental factors, especially when combined, have a negative impact on the iron and iodine status of females.

The aim of this study was to assess the thyroid status of pregnant women on the basis of biochemical indicators and to evaluate the potential risk of developing iodine deficiency as a result of tobacco smoke exposure by assessing the association between urinary thiocyanate levels and the manifestation of iodine deficiency. The study included 219 pregnant women from the town of Plovdiv and Plovdiv District in Southern Bulgaria. The levels of urinary iodine, thyroid-stimulating hormone (TSH), free thyroxine (FT4), and urinary thiocyanate as indicators of tobacco smoke exposure were measured. Most of the pregnant women (60.1 %) were found to have iodine deficiency, 10.6 % of them had TSH values greater than 4 mIU/L, and 16.4 % had FT4 below 9 pmol/L. There were negative correlations between urinary iodine levels and thiocyanate/creatinine ratio (R = -0.148, р = 0.034) and between thiocyanate/creatinine ratio and FT4 (R = -0.379, p < 0.0001); thiocyanate/creatinine ratio and serum TSH were positively correlated (R = 0.169, p = 0.019). Logistic regression analysis showed that pregnant women in whom the thiocyanate/creatinine ratio was greater than the median value of 3.57 mg/g had a 3.882-fold higher risk of developing iodine deficiency (urinary iodine <150 μg/L) than the pregnant women with lower thiocyanate levels (OR = 3.882, 95 % CI 1.402-10.751, p = 0.009). Higher levels of urinary thiocyanate were found in women exposed to tobacco smoke, and quantification of these ions in urine provided a fast non-invasive method to monitor thiocyanate load. Due to the competitive inhibition of iodine intake by thiocyanates, their levels should be carefully monitored, especially in cases of severe iodine deficiency.

Objective: This study highlights the relationship between some eating habits and iron status during pregnancy., Subjects: The study included 219 healthy pregnant women aged 27.6 ± 5.7 years from southern Bulgaria., Methods: Subjects' iron status was assessed on the basis of the following iron indicators: hemoglobin (Hb), serum ferritin (SF), serum transferrin receptor (sTfR), and body iron index (mg/kg)., Results: Severe anemia among the women from southern Bulgaria was not observed. Advanced pregnancy and some eating habits are factors that deteriorate iron status. Women who had consumed fish at least 3 times a week had lower levels of sTfR (р = 0.008), higher levels of SF (р = 0.05), and lower levels of body iron (р = 0.018). Frequent legume consumption was related to increased levels of sTfR (р = 0.036). Pregnant women with a high frequency of coffee consumption had lower values of body iron (р < 0.0001). Women who had consumed cow's milk at least 3 times a week had lower levels of SF (р = 0.026) and body iron (р = 0.042)., Conclusions: Regular consumption of fish and legumes, rarely drinking coffee, and milk consumption during the intervals between food intake are conditions for optimization of iron status during pregnancy.