Browsing by Author "Konrade, Ilze"
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Item Assessment of iodine and selenium nutritional status in women of reproductive age in Latvia(2021-11-05) Veisa, Vija; Kalere, Ieva; Zake, Tatjana; Strele, Ieva; Makrecka, Marina; Upmale-Engela, Sabine; Skesters, Andrejs; Rezeberga, Dace; Lejnieks, Aivars; Pudule, Iveta; Grinberga, Daiga; Velika, Biruta; Dambrova, Maija; Konrade, Ilze; Department of Obstetrics and Gynaecology; Department of Internal Diseases; Institute of Occupational Safety and Environmental Health; Bioķīmijas zinātniskā laboratorijaBackground and Objectives: Adequate dietary intake of iodine and selenium is essential during pregnancy. While iodine is vital for maternal thyroid function and fetal development, selenium contributes to the regulation of thyroid function and thyroid autoimmunity. This study aimed to assess the consumption of iodine-and selenium-containing products by women of reproductive age and the iodine and selenium nutritional status of pregnant women in Latvia. Materials and Methods: Population health survey (2010–2018) data were used to characterize dietary habits in women of reproductive age. Additionally, 129 pregnant women in the first trimester were recruited; they completed a questionnaire and were tested for thyroid function, urinary iodine concentration (UIC), and serum selenium and selenoprotein P levels. Results: The use of some dietary sources of iodine (e.g., milk and dairy products) and selenium (e.g., bread) has decreased in recent years. Less than 10% of respondents reported the use of iodized salt. The use of supplements has become more common (reported by almost 50% of respondents in 2018). Dietary habits were similar in pregnant women, but the use of supplements was even higher (almost 70%). Nevertheless, most supplements used in pregnancy had insufficient contents of iodine and selenium. Thyroid function was euthyreotic in all women, but 13.9% of participants had a thyroid peroxidase antibodies (TPO-ab) level above 60 IU/mL. The median UIC (IQR) was 147.2 (90.0–248.1) µg/gCr, and 52.8% of pregnant women had a UIC below 150 µg/gCr. The mean selenium (SD) level was 101.5 (35.6) µg/L; 30.1% of women had a selenium level below 80 µg/L. The median selenoprotein P level was 6.9 (3.1–9.0) mg/L. Conclusions: Iodine nutrition in Latvian population of pregnant women was near the lower limit of adequate and a third of the population had a selenium deficiency. Supplements were frequently used, but most did not contain the recommended amounts of iodine and selenium.Item Case Report : Micro-RNAs in Plasma From Bilateral Inferior Petrosal Sinus Sampling and Peripheral Blood From Corticotroph Pituitary Neuroendocrine Tumors(2022-04-22) Niedra, Helvijs; Peculis, Raitis; Konrade, Ilze; Balcere, Inga; Romanovs, Mihails; Steina, Liva; Stukens, Janis; Sokolovska, Jelizaveta; Klovins, Janis; Rovite, Vita; Department of Internal DiseasesObjective: Circulating miRNAs are found in bodily fluids including plasma and can serve as biomarkers for diseases. The aim of this study was to provide the first insight into the landscape of circulating miRNAs in close proximity to the adrenocorticotropic hormone (ACTH) secreting PitNET. To achieve this objective next-generation sequencing of miRNAs in plasma from bilateral inferior petrosal sinus sampling (BIPSS) - a gold standard in diagnosing ACTH-secreting PitNETs was carried out and selected miRNA candidates were further tested by RT-qPCR in independent patient cohorts. Methods: Sinistral (left) and dextral (right) BIPSS blood samples of the patient were collected in three time points: before the administration of corticotropin-releasing hormone, 5 and 15 minutes after stimulation. In differential expression analysis, sinistral plasma was compared with dextral. The selected miRNA candidates were tested in plasma by RT-qPCR in two patient groups: 1) in five ACTH secreting PitNET patients with plasma samples taken before and 24 hours after surgery, 2) in 12 ACTH secreting PitNET patients vs. 9 non-functioning PitNET patients. Results: BIPSS concluded that the highest amount of ACTH was released in the sinistral side at the 5th minute mark indicating a presence of a tumor. The highest amount of differentially expressed miRNAs was observed 5 minutes after stimulation (20 upregulated, 14 downregulated). At the 5th minute mark in sinistral plasma, two miRNAs were identified: hsa-miR-7-5p and hsa-miR-375-3p that were highly upregulated compared to other BIPSS samples and peripheral plasma samples. Further testing by qPCR revealed significant reduction of miR-7-5p in plasma 24 hours after surgery and upregulation in plasma of ACTH secreting PitNET patients compared to non-functioning PitNET patients (P =0.0013). Conclusions: By stimulating the ACTH secreting PitNET with CRH a rapid increase of two miRNAs (hsa-mir-7-5p, hsa-mir-375-3p) and ACTH can be observed in sinistral inferior petrosal (tumor side). A decrease of miR-7-5p in plasma after surgery and upregulation in plasma of ACTH secreting PitNET patients was discovered implying that further studies of this miRNA as diagnostic marker is needed.Item Consumption of thyroid medications as an indicator of increase of thyroid morbidity in Latvia from 2011 to 2014(2019-08-01) Kalere, Ieva; Strele, Ieva; Miglinieks, Martiņš; Repša, Ilze; Pildava, Santa; Romanovs, Mihails; Pirags, Valdis; Konrade, Ilze; Rīga Stradiņš UniversityThe most common autoimmune disorders with clinically opposite manifestations are hypothyroidism in Hashimoto's thyroiditis and hyperthyroidism in Graves' disease. The healthcare burden of thyroid disease is substantial, resulting in substantial health care costs. The aim of the present analysis is to assess the use of thyroid medications in Latvia from 2011 to 2014 by age and gender. Our study used reimbursed medication prescriptions data, collected by the National Health Service of Latvia. The main indicator was the number of prevalent users of thyroid medications each year from 2011 to 2014, stratified by age and gender. From 2011 to 2014, the number of thyroxine users per 100 000 revealed a statistically significant increase in all age and gender groups, except in 0- to 9-year-old girls. The number of Thiamazole users among men increased in the age group from 40 to 89 years and in women age groups above 49 years. Increasing sales of both thyroid hormones and antithyroid medications are also observed in Estonia and Lithuania, indicating that growing thyroid morbidity is an issue in the whole region. The substantial increase in number of patients highlights the necessity for national guidelines on the use of thyroid function tests and standards of medical care.Item A cross-sectional survey of urinary iodine status in Latvia(2014) Konrade, Ilze; Neimane, Lolita; Makrecka, Marina; Strele, Ieva; Liepinsh, Edgars; Lejnieks, Aivars; Vevere, Parsla; Gruntmanis, Ugis; Pirags, Valdis; Dambrova, Maija; Rīga Stradiņš UniversityBackground and objective: A nationwide survey of schoolchildren was conducted to detect regional differences in urinary iodine excretion in Latvia and to compare the results with data from the newborn thyroid-stimulating hormone (TSH) screening database as well with the results of a similar study performed in Latvia 10 years ago. Materials and methods: We conducted a cross-sectional school-based cluster survey of 915 children aged 9-12 years in 46 randomly selected schools in all regions of Latvia. Urine samples, questionnaires on the consumption of iodized salt and information on socioeconomic status were collected. TSH levels in newborns were also measured. Results: The median creatinine-standardized urinary iodine concentration (UIC) in our study was 107.3 mg/g Cr. UIC measurements indicative of mild iodine deficiency were present in 31.6%, moderate deficiency in 11.9% and severe deficiency in 2.8% of the participants. The prevalence of iodine deficiency was the highest in the southeastern region of Latgale and the northeastern region of Vidzeme. The prevalence of TSH values >5 mIU/L followed a similar pattern. The self-reported prevalence of regular iodized salt consumption was 10.2%. Children from urban schools had a significantly lower UIC than children from rural schools. Conclusions: Our findings suggest that although the overall median UIC in Latvian schoolchildren falls within the lower normal range, almost 50% of the schoolchildren are iodine deficient, especially in urban schools and in the eastern part of Latvia. The absence of amandatory salt iodization program puts a significant number of children and pregnantwomen at risk.Item Erratum : Methylglyoxal modification of Na v 1.8 facilitates nociceptive neuron firing and causes hyperalgesia in diabetic neuropathy (Nature Medicine (2012) 18 (926-933))(2012-09) Bierhaus, Angelika; Fleming, Thomas; Stoyanov, Stoyan; Leffler, Andreas; Babes, Alexandru; Neacsu, Cristian; Sauer, Susanne K.; Eberhardt, Mirjam; Schnölzer, Martina; Lasischka, Felix; Neuhuber, Winfried L.; Kichko, Tatjana I.; Konrade, Ilze; Elvert, Ralf; Mier, Walter; Pirags, Valdis; Lukic, Ivan K.; Morcos, Michael; Dehmer, Thomas; Rabbani, Naila; Thornalley, Paul J.; Edelstein, Diane; Nau, Carla; Forbes, Josephine; Humpert, Per M.; Schwaninger, Markus; Ziegler, Dan; Stern, David M.; Cooper, Mark E.; Haberkorn, Uwe; Brownlee, Michael; Reeh, Peter W.; Nawroth, Peter P.; Department of Doctoral StudiesItem Evaluation of the Possibility to Detect Circulating Tumor DNA From Pituitary Adenoma(2019-09-18) Megnis, Kaspars; Peculis, Raitis; Rovite, Vita; Laksa, Pola; Niedra, Helvijs; Balcere, Inga; Caune, Olivija; Breiksa, Austra; Nazarovs, Jurijs; Stukens, Janis; Konrade, Ilze; Pirags, Valdis; Klovins, Janis; Department of Internal DiseasesObjective: Circulating free DNA (cfDNA) in general and circulating tumor DNA (ctDNA) in particular is becoming an increasingly used form of liquid biopsy biomarkers. In this study, we are investigating the ability to detect ctDNA from the plasma of pituitary adenoma (PA) patients. Design: Tumor tissue samples were obtained from planed PA resections, before which blood plasma samples were taken. Somatic variants found in PA tissue samples were evaluated in related cfDNA, isolated from plasma samples. Methods: Sanger sequencing, as well as previously obtained whole-exome sequencing data, were used to evaluate somatic variants composition in tumor tissue samples. cfDNA was isolated from the same PA patients and competitive allele-specific TaqMan PCR and amplicon-based next-generation sequencing (NGS) approach were used for targeted detection of variants found in corresponding tumor tissue samples. Results: Using NGS-based analysis, we detected five out of 17 somatic variants in 40 to 60% of total reads, three variants in 0.50–5.00% of total read count, including GNAS c.601C>T, which was detected using ultra-deep NGS (1.78 million X) in 0.77% of amplicons reads. Nine variants were not detected. We also detected We were not able to detect variant found in PA tissue in cfDNA using cast-PCR, indicating that the portion of variant-containing ctDNA in total isolated cfDNA is too small to be detected with this method. Conclusions: For the first time, we demonstrate the possibility to detect somatic variants of PA in cfDNA isolated from patients' blood plasma. Whether the source of variant detected in cfDNA is PA should be further tested.Item Fasting-Mimicking Diet Reduces Trimethylamine N-Oxide Levels and Improves Serum Biochemical Parameters in Healthy Volunteers(2022-03-01) Videja, Melita; Sevostjanovs, Eduards; Upmale-Engela, Sabine; Liepinsh, Edgars; Konrade, Ilze; Dambrova, Maija; Faculty of Pharmacy; Department of Internal DiseasesElevated plasma levels of trimethylamine N-oxide (TMAO) have been proposed as a diet-derived biomarker of cardiometabolic disease risk. Caloric restriction is the most common dietary intervention used to improve cardiometabolic health; however, novel trends suggest a fasting-mimicking diet (FMD) as a more feasible alternative. FMD is a variation of intermittent fasting, based on caloric restriction and limitation of protein sources of animal origin, applied in daily cycles during a 5-day period. As TMAO is intensively produced by gut microbiota after the consumption of animal-derived products, we aim to investigate whether a 5-day FMD affects plasma TMAO levels and markers of metabolic health. To investigate whether an increase in vegetable intake possesses similar effects on TMAO levels and metabolic parameters, healthy volunteers (n = 24) were subjected to a 5-day FMD and 19 volunteers served as a reference group (VEG). This group of volunteers consumed an additional four servings of vegetables per day, but otherwise stayed on their usual diet. FMD resulted in a twofold decrease in plasma TMAO levels, which was not evident in the volunteers from the VEG group. Moreover, FMD led to a weight loss of 2.8 ± 0.2 kg and a subsequent reduction in BMI compared to baseline. The FMD group exhibited a significant elevation in plasma ketone bodies (14-fold compared to baseline) and a decrease in IGF-1 levels by 37 ± 8 ng/mL. Since fasting glucose and C-peptide levels decreased, all volunteers in the FMD group showed improved insulin sensitivity and a decreased HOMA-IR index. In contrast, in the VEG group, only a slight reduction in plasma levels of fasting glucose and triglycerides was noted. In conclusion, we show that FMD is a viable strategy to reduce plasma levels of TMAO by limiting caloric intake and animal-derived protein consumption. The reduction in the level of TMAO could be an additional benefit of FMD, leading to a reduced risk of cardiometabolic diseases.Item Genome wide analysis of circulating miRNAs in growth hormone secreting pituitary neuroendocrine tumor patients’ plasma(2022-09-09) Niedra, Helvijs; Peculis, Raitis; Litvina, Helena Daiga; Megnis, Kaspars; Madrika, Ilona; Balcere, Inga; Romanovs, Mihails; Steina, Liva; Stukens, Janis; Breiksa, Austra; Nazarovs, Jurijs; Sokolovska, Jelizaveta; Liutkeviciene, Rasa; Vilkevicute, Alvita; Konrade, Ilze; Rovite, Vita; Department of Internal DiseasesBackground: Circulating plasma miRNAs have been increasingly studied in the field of pituitary neuroendocrine tumor (PitNET) research. Our aim was to discover circulating plasma miRNAs species associated with growth hormone (GH) secreting PitNETs versus assess how the plasma levels of discovered miRNA candidates are impacted by SSA therapy and whether there is a difference in their levels between GH secreting PitNETs versus other PitNET types and healthy individuals. Design: We compared plasma miRNA content and levels before and after surgery focusing on GH secreting PitNET patients. Selected miRNA candidates from our data and literature were then tested in a longitudinal manner in somatostatin analogues (SSA) treatment group. Additionally, we validated selected targets in an independent GH secreting PitNET group. Methods: miRNA candidates were discovered using the whole miRNA sequencing approach and differential expression analysis. Selected miRNAs were then analyzed using real-time polymerase chain reaction (qPCR). Results: Whole miRNA sequencing discovered a total of 16 differentially expressed miRNAs (DEMs) in GH secreting PitNET patients’ plasma 24 hours after surgery and 19 DEMs between GH secreting PitNET patients’ plasma and non-functioning (NF) PitNET patients’ plasma. Seven miRNAs were selected for further testing of which miR-625-5p, miR-503-5p miR-181a-2-3p and miR-130b-3p showed a significant downregulation in plasma after 1 month of SSA treatment. mir-625-5p was found to be significantly downregulated in plasma of GH secreting PitNET patients vs. NF PitNET patients. miR-625-5p alongside miR-130b-3p were also found to be downregulated in GH PitNETs compared to healthy individuals. Conclusions: Our study suggests that expression of plasma miRNAs miR-625-5p, miR-503-5p miR-181a-2-3p and miR-130b-3p in GH secreting PitNETs is affected by SSA treatment. Additionally, miR-625-5p can distinguish GH secreting PitNETs from other PitNET types and healthy controls warranting further research on these miRNAs for treatment efficacy.Item Heterogeneity of tissue IL-17 and tight junction proteins expression demonstrated in patients with autoimmune thyroid diseases(2018-06-01) Zake, Tatjana; Skuja, Sandra; Kalere, Ieva; Konrade, Ilze; Groma, Valerija; Department of Internal Diseases; Institute of Anatomy and AnthropologyTh17 cells together with their hallmark cytokine interleukin (IL)-17 were identified as crucial contributing factors in the pathogenesis of thyroid autoimmunity. The cytokine-regulated tight junction (Tj) disruption is thought to be essential in the initiation and/or development of several diseases. Still, the role of IL-17 maintaining Tj integrity in autoimmune thyroid diseases (AITDs) has not yet been evaluated. We aimed to investigate integrity of the thyroid follicle by studying immunoexpression of cellular Tj - zonula occludens (ZO)-1 and claudin-1 proteins coupled to IL-17A and CD68 detection in AITD patients compared with controls. Thirty-five adult patients undergoing thyroidectomy and presenting 18 cases of Hashimoto thyroiditis (HT), 7 of Graves' disease (GD) as well as 10 subjects of colloid goiter without autoimmune component served as controls were enrolled in this study. An immunohistochemical analysis including IL-17A, ZO-1, claudin-1, and CD68 detection was performed in each case. The correlation of IL-17A with Tj and CD68 in patients with AITD was also analyzed. Apart from inflammatory cells, we evidenced a stronger expression level of IL17A in the thyroid follicular cells in HT patients when compared with GD or colloid goiter. A significant reduction of ZO-1 immunoreactivity was observed in the thyrocytes in HT patients, whereas no significant differences were found in claudin-1 expression in HT and GD compared with colloid goiter patients. A significantly higher number of thyroid follicles with CD68-positive cells was found in HT patients than that in patients with GD or colloid goiter. In HT patients, the expression of IL-17A in the follicular cells was positively correlated with CD68 immunopositivity, whereas no association with claudin-1 or ZO-1 expression was found. GD patients did not reveal any significant correlation of IL-17A with Tj and CD68. Strong overexpression of IL-17A observed in the thyroid epithelial cells is associated with the presence of intrafollicular CD68-positive cells in HT patients. We evidenced the changes in molecules of thyrocyte junctional complexes highlighting impairment of the thyroid follicle integrity in HT, but no association with IL-17A was found.Item Immunological mechanisms of autoimmune thyroid diseases : A shift in the traditional TH1/TH2 paradigm(2019-05-01) Zaķe, Tatjana; Skuja, Sandra; Lejnieks, Aivars; Groma, Valerija; Konrade, Ilze; Institute of Anatomy and Anthropology; Department of Internal DiseasesAutoimmune thyroid diseases (AITD) mainly include Hashimoto's thyroiditis (HT) and Graves' disease (GD), which are characterised by the presence of circulating antibodies against various thyroid autoantigens and infiltration of the thyroid gland by autoreactive lymphocytes. Despite the significant advancement in the knowledge of AITD pathogenesis in the last decade, the specific immunological mechanisms responsible for development of the disease are not thoroughly understood. Classically, HT has long been considered as a T helper (Th)1-mediated disease, while a Th2-driven autoimmune response is dominant for GD development. However, this classification has changed due to the description of Th17 lymphocytes, which suggested participation of these cells in AITD, particularly HT pathogenesis. Moreover, a shift in the balance between Th17 and T regulatory (Treg) cells has been observed in thyroid autoimmunity. We have observed overexpression of IL-17, the prominent effector cytokine of Th17, within thyroid tissues from HT and GD patients in our studies. The present review will focus on recent data regarding the role of Treg and Th17 lymphocytes in AITD pathogenesis. In addition, the impact and proposed mechanisms of the predominant environmental factors triggering the autoimmune response to the thyroid will be discussed.Item Iodine deficiency in Latvia : Current status and need for national recommendations(2017-12) Konrade, Ilze; Kalere, Ieva; Strele, Ieva; Makrecka-Kuka, Marina; Veisa, Vija; Gavars, Didzis; Rezeberga, Dace; Pirags, Valdis; Lejnieks, Aivars; Gruntmanis, Ugis; Neimane, Lolita; Liepiņš, Edgars; Dambrova, Maija; Rīga Stradiņš UniversityIn the absence of a mandatory salt iodisation programme, two nationwide cross-sectional cluster surveys revealed persisting iodine deficiency among Latvian schoolchildren during the spring season and a noteworthy iodine deficiency in pregnant women in Latvia; these deficiencies warrant intervention. The consequences of mild-to-moderate iodine deficiency during pregnancy and lactation can adversely affect foetal brain development. Data from a Latvian population survey revealed the consumption of approximately 100 μg of iodine per day through foods and iodised salt. Therefore, strategies to increase the consumption of iodine-containing products should be implemented, particularly for children. In addition, to meet the increased iodine requirement during pregnancy, pregnant women should take daily supplements containing 150 μg iodine from the earliest time possible. All women of childbearing age should be advised to increase their dietary iodine intake by using iodised table salt and iodine-rich products: seafood, milk and milk products. For women with pre-existing thyroid pathologies, the medical decision should be considered on a case-by-case basis. Urinary iodine concentration monitoring among schoolchildren and pregnant women and neonatal thyrotropin registry analysis every five years would be an appropriate strategy for maintaining iodine intake within the interval that prevents iodine deficiency disorders.Item Medication for Acromegaly Reduces Expression of MUC16, MACC1 and GRHL2 in Pituitary Neuroendocrine Tumour Tissue(2021-02-15) Saksis, Rihards; Silamikelis, Ivars; Laksa, Pola; Megnis, Kaspars; Peculis, Raitis; Mandrika, Ilona; Rogoza, Olesja; Petrovska, Ramona; Balcere, Inga; Konrade, Ilze; Steina, Liva; Stukens, Janis; Breiksa, Austra; Nazarovs, Jurijs; Sokolovska, Jelizaveta; Pirags, Valdis; Klovins, Janis; Rovite, Vita; Rīga Stradiņš UniversityAcromegaly is a disease mainly caused by pituitary neuroendocrine tumor (PitNET) overproducing growth hormone. First-line medication for this condition is the use of somatostatin analogs (SSAs), that decrease tumor mass and induce antiproliferative effects on PitNET cells. Dopamine agonists (DAs) can also be used if SSA treatment is not effective. This study aimed to determine differences in transcriptome signatures induced by SSA/DA therapy in PitNET tissue. We selected tumor tissue from twelve patients with somatotropinomas, with half of the patients receiving SSA/DA treatment before surgery and the other half treatment naive. Transcriptome sequencing was then carried out to identify differentially expressed genes (DEGs) and their protein–protein interactions, using pathway analyses. We found 34 upregulated and six downregulated DEGs in patients with SSA/DA treatment. Three tumor development promoting factors MUC16, MACC1, and GRHL2, were significantly downregulated in therapy administered PitNET tissue; this finding was supported by functional studies in GH3 cells. Protein–protein interactions and pathway analyses revealed extracellular matrix involvement in the antiproliferative effects of this type of the drug treatment, with pronounced alterations in collagen regulation. Here, we have demonstrated that somatotropinomas can be distinguished based on their transcriptional profiles following SSA/DA therapy, and SSA/DA treatment does indeed cause changes in gene expression. Treatment with SSA/DA significantly downregulated several factors involved in tumorigenesis, including MUC16, MACC1, and GRHL2. Genes that were upregulated, however, did not have a direct influence on antiproliferative function in the PitNET cells. These findings suggested that SSA/DA treatment acted in a tumor suppressive manner and furthermore, collagen related interactions and pathways were enriched, implicating extracellular matrix involvement in this anti-tumor effect of drug treatment.Item Melatonin concentrations and sleep quality in patients with type 2 diabetes and obesity(2019-05-01) Kalere, Ieva; Konrade, Ilze; Proskurina, Anna; Upmale, Sabine; Zaķe, Tatjana; Limba, Normunds; Krieviņa, Gita; Lejnieks, Aivars; Tretjakovs, Peteris; Department of Human Physiology and Biochemistry; Department of Internal DiseasesThere is a close relationship between melatonin as a circadian regulator and insulin, glucagon and somatostatin production. This study aimed to describe subgroups of type 2 diabetes mellitus (T2DM) patients that may benefit from melatonin clock-targeting properties. The study involved 38 participants: 26 T2DM patients, and 12 participants without diabetes in the control group. Subjects were asked to complete the questionnaire of Pittsburgh Sleep Quality Index (PSQI). Standard biochemical venous sample testing was performed, and a sample of saliva was collected for melatonin testing. Melatonin concentration in participants without obesity (body mass index (BMI) < 30 kg/m 2 ) was significantly higher than in obese participants: 13.2 (6.4; 23.50) pg/ml vs 5.9 (0.78; 13.1) pg/ml, p = 0.035. Subjects with BMI 30 kg/m 2 had a significantly higher PSQI score than non-obese subjects: 7 (4.5; 10) vs 5.5 (3; 7), p = 0.043. T2DM patients showed significantly lower levels of melatonin than the control group: 6.1 (0.78; 12.2) pg/ml vs 17.8 (8.2; 25.5) pg/ml, p = 0.003. T2DM patients using short-acting insulin analogues showed a significantly higher PSQI score than patients not using insulin: 9 (6; 10) vs 6 (3; 8), respectively (p = 0.025). Poor sleep quality was more prevalent in patients with diabetic retinopathy than in those without this complication (p = 0.031). Lower melatonin levels were detected in T2DM and obese patients. Furthermore, poor sleep quality was observed in T2DM patients using short-acting insulin analogues and those with diabetic retinopathy, and obese individuals.Item Metformin strongly affects transcriptome of peripheral blood cells in healthy individuals(2019-11-01) Ustinova, Monta; Silamikelis, Ivars; Kalnina, Ineta; Ansone, Laura; Rovite, Vita; Elbere, Ilze; Radovica-Spalvina, Ilze; Fridmanis, Davids; Aladyeva, Jekaterina; Konrade, Ilze; Pirags, Valdis; Klovins, Janis; Rīga Stradiņš UniversityMetformin is a commonly used antihyperglycaemic agent for the treatment of type 2 diabetes mellitus. Nevertheless, the exact mechanisms of action, underlying the various therapeutic effects of metformin, remain elusive. The goal of this study was to evaluate the alterations in longitudinal whole-blood transcriptome profiles of healthy individuals after a one-week metformin intervention in order to identify the novel molecular targets and further prompt the discovery of predictive biomarkers of metformin response. Next generation sequencing-based transcriptome analysis revealed metformin-induced differential expression of genes involved in intestinal immune network for IgA production and cytokine-cytokine receptor interaction pathways. Significantly elevated faecal sIgA levels during administration of metformin, and its correlation with the expression of genes associated with immune response (CXCR4, HLA-DQA1, MAP3K14, TNFRSF21, CCL4, ACVR1B, PF4, EPOR, CXCL8) supports a novel hypothesis of strong association between metformin and intestinal immune system, and for the first time provide evidence for altered RNA expression as a contributing mechanism of metformin’s action. In addition to universal effects, 4 clusters of functionally related genes with a subject-specific differential expression were distinguished, including genes relevant to insulin production (HNF1B, HNF1A, HNF4A, GCK, INS, NEUROD1, PAX4, PDX1, ABCC8, KCNJ11) and cholesterol homeostasis (APOB, LDLR, PCSK9). This inter-individual variation of the metformin effect on the transcriptional regulation goes in line with well-known variability of the therapeutic response to the drug.Item Mildronate treatment alters γ-butyrobetaine and l -carnitine concentrations in healthy volunteers(2011-09) Liepinsh, Edgars; Konrade, Ilze; Skapare, Elina; Pugovics, Osvalds; Grinberga, Solveiga; Kuka, Janis; Kalvinsh, Ivars; Dambrova, Maija; Rīga Stradiņš UniversityObjectives In this study, we aimed to investigate the effects of long-term administration of the cardioprotective drug mildronate on the concentrations of l-carnitine and γ-butyrobetaine in healthy volunteers. Methods Mildronate was administered perorally, at a dosage of 500 mg, twice daily. Plasma and urine samples were collected weekly. Daily meat consumption within an average, non-vegetarian diet was monitored. l-Carnitine, γ-butyrobetaine and mildronate concentrations were measured using the UPLC/MS/MS method. Key findings After 4 weeks, the average concentrations of l-carnitine in plasma significantly decreased by 18%. The plasma concentrations of γ-butyrobetaine increased about two-fold, and this effect was statistically significant in both the male and female groups. In urine samples, a significant increase in l-carnitine and γ-butyrobetaine levels was observed, which provides evidence for increased excretion of both substances during the mildronate treatment. At the end of the treatment period, the plasma concentration of mildronate was 20 μm on average. There were no significant differences between the effects observed in female and male volunteers. Meat consumption partially reduced the l-carnitine-lowering effects induced by mildronate. Conclusions Long-term administration of mildronate significantly lowers l-carnitine plasma concentrations in non-vegetarian, healthy volunteers.Item Milk as an essential source of iodine in Latvian population(2017-12) Neimane, Lolita Vija; Konrade, Ilze; Avotiņa, Gita; Klaviņa, Aneka; Zagorska, Jeļena; Martinsone, Inese; Cauce, Vinita; Kalere, Ieva; Baylon, Vincenzo; Lejnieks, Aivars; Department of Sports and Nutrition; Department of Internal Diseases; Laboratory of Hygiene and Occupational Diseases; Department of PhysicsMilk and dairy products are studied as alternative iodine sources, because salt iodisation is controversial due to high salt consumption leading to cardiovascular diseases. However, the iodine concentration in milk markedly varies. This study evaluated the iodine concentration in cow's milk available in the Latvian market. Iodine and fat concentration was analysed with a spectrophotometer "Varian Cary 50" based ISO 2446:2008 in 20 milk samples. Data from the Central Statistical Bureau and survey among pregnant women were used to analyse milk product consumption and its impact on iodine status. Average iodine concentration in milk samples was 457.6 (179.6) μg/L, winter samples had a higher concentration of iodine than summer samples: 563.4 (329.6) μg/L and 469.2 (162.0) μg/L, but this is not statistically significant p < 0.05. Iodine concentration in skimmed milk was 490 μg/L, milk with the reduced fat content 501.7 (174.8) μg/L, and whole milk - 422.6 (192.1)1 μg/L. Milk consumption decreased from 2002 to 2014, while yogurt and cheese consumption increased. Higher consumption of milk and milk products was related to higher urinary iodine concentration ρ = 0.115; p = 0.003. Milk and milk products are an important iodine source in Latvia and their consumption should be promoted.Item Novel susceptibility loci identified in a genome-wide association study of type 2 diabetes complications in population of Latvia(2021) Ustinova, Monta; Peculis, Raitis; Rescenko, Raimonds; Rovite, Vita; Zaharenko, Linda; Elbere, Ilze; Silamikele, Laila; Konrade, Ilze; Sokolovska, Jelizaveta; Pirags, Valdis; Klovins, Janis; Faculty of MedicineBackground: Type 2 diabetes complications cause a serious emotional and economical burden to patients and healthcare systems globally. Management of both acute and chronic complications of diabetes, which dramatically impair the quality of patients' life, is still an unsolved issue in diabetes care, suggesting a need for early identification of individuals with high risk for developing diabetes complications. Methods: We performed a genome-wide association study in 601 type 2 diabetes patients after stratifying them according to the presence or absence of four types of diabetes complications: diabetic neuropathy, diabetic nephropathy, macrovascular complications, and ophthalmic complications. Results: The analysis revealed ten novel associations showing genome-wide significance, including rs1132787 (GYPA, OR = 2.71; 95% CI = 2.02–3.64) and diabetic neuropathy, rs2477088 (PDE4DIP, OR = 2.50; 95% CI = 1.87–3.34), rs4852954 (NAT8, OR = 2.27; 95% CI = 2.71–3.01), rs6032 (F5, OR = 2.12; 95% CI = 1.63–2.77), rs6935464 (RPS6KA2, OR = 2.25; 95% CI = 6.69–3.01) and macrovascular complications, rs3095447 (CCDC146, OR = 2.18; 95% CI = 1.66–2.87) and ophthalmic complications. By applying the targeted approach of previously reported susceptibility loci we managed to replicate three associations: MAPK14 (rs3761980, rs80028505) and diabetic neuropathy, APOL1 (rs136161) and diabetic nephropathy. Conclusions: Together these results provide further evidence for the implication of genetic factors in the development of type 2 diabetes complications and highlight several potential key loci, able to modify the risk of developing these conditions. Moreover, the candidate variant approach proves a strong and consistent effect for multiple variants across different populations.Item Novel variant of the androgen receptor gene in a patient with complete androgen insensitivity syndrome and polyorchidism(2019) Konrade, Ilze; Zavorikina, Julija; Fridvalde, Aija; Rots, Dmitrijs; Kalere, Ieva; Strumfa, Ilze; Dambrova, Maija; Gailite, Linda; Department of Internal Diseases; Scientific Laboratory of Molecular Genetics; Faculty of Pharmacy; Department of Pathology; Department of Human Physiology and BiochemistryIntroduction: Complete androgen insensitivity (CAIS) in 65–95% cases is caused by pathogenic allelic variants (mutations) in the gene encoding androgen receptor (AR gene) and is characterized by female phenotype development with a male karyotype (46, XY). Patients are usually diagnosed during puberty and undergo gonadectomy due to increased testicular germ cell tumor risk. Only a few outcomes have been reported in older individuals with postponed gonadectomy. Case presentation: A 48-year-old CAIS patient presented with polyorchidism (four testes) without gonadal malignancies. Genetic testing identified a novel allelic variant in the AR gene [c.2141T>G (p.Phe805Cys)] causing the clinical symptoms. Conclusion: We have described a unique patient with CAIS and polyorchidism without malignancies in her late 40's bearing a novel likely pathogenic variant in the AR gene.Item Plasma circulating microRNAs in patients with stable coronary artery disease - Impact of different cardiovascular risk profiles and glomerular filtration rates(2021-04-22) Trusinskis, Karlis; Lapsovs, Maris; Paeglite, Sandra; Knoka, Evija; Caunite, Laima; Mazule, Mairita; Briede, Ieva; Jegere, Sanda; Kumsars, Indulis; Narbute, Inga; Konrade, Ilze; Erglis, Andrejs; Lejnieks, Aivars; Department of Internal DiseasesBackground and Aim: Plasma circulating microRNA (miRNA)-126, -145, and -155 are associated with vascular remodeling, atherosclerotic lesion formation, and plaque vulnerability. In this study, we evaluated the levels of plasma circulating miRNAs in patients with stable coronary artery disease (CAD), different cardiovascular risk profiles, and different glomerular filtration rates (GFR). Methods and Results: Forty patients with stable CAD admitted for elective percutaneous coronary intervention (PCI) were enrolled in a prospective study. Before PCI, fasting blood samples were obtained to evaluate clinical parameters and miRNA-126 and miRNA-155 expression. The GFR was calculated by the MDRD and CKD-EPI formulas, and the severity of CAD was calculated according to the SYNTAX score. All these parameters were correlated with miRNAs. The association between miRNA levels and clinical characteristics was evaluated. The expression of miRNA-126 positively correlated with a higher SYNTAX score (r = 0.337; p=0.034); however, no significant correlations between miR-126, GFR, and clinical characteristics were observed. Higher plasma levels of miRNA-155 correlated with increased levels of triglycerides (r = 0.317; P = 0.049), C-peptide (r = 0.452; P = 0.011), and the HOMA index (r = 0.447; P = 0.012) and a higher body mass index (BMI) (r = 0.385; P = 0.015). GFR and miRNA-155 (MDRD - Rho=0.353; P = 0.027. CKD-EPI - Rho=0.357; P = 0.026) were found to have a moderate correlation, although miRNA-155 had no correlation with the SYNTAX score. Conclusion: Plasma circulating miRNA-126 levels were increased in patients with severe atherosclerosis as determined by the SYNTAX score. Elevated miRNA-155 expression was observed in patients with Stage 1 GFR but was lower in patients with Stages 2 and 3 GFR. Plasma circulating miRNA-155 had positive correlations with higher levels of BMI, HOMA index, C-peptide, and triglycerides. Relevance for Patients: Although further investigations are needed to confirm the role of miRNA-155 and miRNA-126, they may serve as potential biomarkers detecting severity of CAD, lowering of kidney function and metabolic syndrome.Item Significantly altered peripheral blood cell DNA methylation profile as a result of immediate effect of metformin use in healthy individuals(2018-12-13) Elbere, Ilze; Silamikelis, Ivars; Ustinova, Monta; Kalnina, Ineta; Zaharenko, Linda; Peculis, Raitis; Konrade, Ilze; Ciuculete, Diana Maria; Zhukovsky, Christina; Gudra, Dita; Radovica-Spalvina, Ilze; Fridmanis, Davids; Pirags, Valdis; Schiöth, Helgi B.; Klovins, JanisBackground: Metformin is a widely prescribed antihyperglycemic agent that has been also associated with multiple therapeutic effects in various diseases, including several types of malignancies. There is growing evidence regarding the contribution of the epigenetic mechanisms in reaching metformin's therapeutic goals; however, the effect of metformin on human cells in vivo is not comprehensively studied. The aim of our study was to examine metformin-induced alterations of DNA methylation profiles in white blood cells of healthy volunteers, employing a longitudinal study design. Results: Twelve healthy metformin-naïve individuals where enrolled in the study. Genome-wide DNA methylation pattern was estimated at baseline, 10 h and 7 days after the start of metformin administration. The whole-genome DNA methylation analysis in total revealed 125 differentially methylated CpGs, of which 11 CpGs and their associated genes with the most consistent changes in the DNA methylation profile were selected: POFUT2, CAMKK1, EML3, KIAA1614, UPF1, MUC4, LOC727982, SIX3, ADAM8, SNORD12B, VPS8, and several differentially methylated regions as novel potential epigenetic targets of metformin. The main functions of the majority of top-ranked differentially methylated loci and their representative cell signaling pathways were linked to the well-known metformin therapy targets: regulatory processes of energy homeostasis, inflammatory responses, tumorigenesis, and neurodegenerative diseases. Conclusions: Here we demonstrate for the first time the immediate effect of short-term metformin administration at therapeutic doses on epigenetic regulation in human white blood cells. These findings suggest the DNA methylation process as one of the mechanisms involved in the action of metformin, thereby revealing novel targets and directions of the molecular mechanisms underlying the various beneficial effects of metformin. Trial registration: EU Clinical Trials Register, 2016-001092-74. Registered 23 March 2017, https://www.clinicaltrialsregister.eu/ctr-search/trial/2016-001092-74/LV.