Data mining for the identification of metabolic syndrome status

Metabolic syndrome (MS) is a condition associated with metabolic abnormalities that are characterized by central obesity (e.g. waist circumference or body mass index), hypertension (e.g. systolic or diastolic blood pressure), hyperglycemia (e.g. fasting plasma glucose) and dyslipidemia (e.g. triglyceride and high-density lipoprotein cholesterol). It is also associated with the development of diabetes mellitus (DM) type 2 and cardiovascular disease (CVD). Therefore, the rapid identification of MS is required to prevent the occurrence of such diseases. Herein, we review the utilization of data mining approaches for MS identification. Furthermore, the concept of quantitative population-health relationship (QPHR) is also presented, which can be defined as the elucidation/ understanding of the relationship that exists between health parameters and health status. The QPHR modeling uses data mining techniques such as artificial neural network (ANN), support vector machine (SVM), principal component analysis (PCA), decision tree (DT), random forest (RF) and association analysis (AA) for modeling and construction of predictive models for MS characterization. The DT method has been found to outperform other data mining techniques in the identification of MS status. Moreover, the AA technique has proved useful in the discovery of in-depth as well as frequently occurring health parameters that can be used for revealing the rules of MS development. This review presents the potential benefits on the applications of data mining as a rapid identification tool for classifying MS

A comprehensive medical decision–support framework based on a heterogeneous ensemble classifier for diabetes prediction

Funding Information: Funding: This work was supported by National Research Foundation of Korea-Grant funded by the Korean Government (Ministry of Science and ICT)-NRF-2017R1A2B2012337). Funding Information: This work was supported by National Research Foundation of Korea-Grant funded by the Korean Government (Ministry of Science and ICT)-NRF-2017R1A2B2012337).Peer reviewe

Data mining in biomedicine : current applications and further directions for research

Author name used in this manuscript: S. K. KwokAuthor name used in this manuscript: A. H. C. Tsang2009-2010 > Academic research: refereed > Publication in refereed journalAccepted ManuscriptPublishe

Understanding the Effect of Energy Density and Formulation Factors on the Printability and Characteristics of SLS Irbesartan Tablets-Application of the Decision Tree Model

Selective laser sintering (SLS) is a rapid prototyping technique for the production of three-dimensional objects through selectively sintering powder-based layer materials. The aim of the study was to investigate the effect of energy density (ED) and formulation factors on the printability and characteristics of SLS irbesartan tablets. The correlation between formulation factors, ED, and printability was obtained using a decision tree model with an accuracy of 80%. FT-IR results revealed that there was no interaction between irbesartan and the applied excipients. DSC results indicated that irbesartan was present in an amorphous form in printed tablets. ED had a significant influence on tablets' physical, mechanical, and morphological characteristics. Adding lactose mon-ohydrate enabled faster drug release while reducing the possibility for printing with different laser speeds. However, formulations with crospovidone were printable with a wider range of laser speeds. The adjustment of formulation and process parameters enabled the production of SLS tablets with hydroxypropyl methylcellulose with complete release in less than 30 min. The results suggest that a decision tree could be a useful tool for predicting the printability of pharmaceutical formulations. Tailoring the characteristics of SLS irbesartan tablets by ED is possible; however, it needs to be governed by the composition of the whole formulation

Papers on Anthropology XX

-M. Toomsalu, A. Arend. Medical Collections in Tartu Old Anatomical Theatre – from August Rauber’s anatomy museum to a multifunctional research and edutainment centre -H. Kaarma. A short overview of the work of anthropologists of the Old Anatomical Theatre -M. Viikmaa. Leiu Heapost – 75 -R. Allmäe. Human bones in Salme I boat-grave, the Island of Saaremaa; Estonia -N. Balciuniene, D. graf von Keyserlingk, A. Valanciute, I. Balnyte,A. Macas, A. Tamasauskas. Changes of the chicken chorioallantoic membrane and the behaviour of the transplanted glioblastoma -A. Barzdina, M. Pilmane, A. Petersons. Grap and NF expression in brain tissue in children and adults after fatal traumatic brain injury -D. Batulevicius, G. Skripkiene, V. Batuleviciene, V. Skripka, A. Dabuzinskiene, D. H. Pauza. Complexity of the frog intracardiac neurons. Intracellular injection study -A. Dabuzinskiene, A. Ratkevicius, D. Batulevicius, G. Skripkiene, V. Skripka, G. Liutkiene, U. A. Bagdonas, S. Razbadauskiene. Gender differences in the human cervicothoracic ganglia -C. Dogan, C. Raschka. Anthropometrical and sport constitutional comparison of German male soccer players and male students of sport sciences -D. Gudiene, I. Balnyte, J. Palubinskiene, A. Valanciute. Age related structural changes in human basilar artery -L. Heapost. Taste sensitivity to PTC and colour blindness in Estonians -P. Hussar, I. Tokin, G. Filimonova, I. Tokin, Ü. Hussar. Dexamethazone-induced T-lymphocyte apoptosis in different lymphoid organs -L. Kalichman, E. Kobyliansky. Hand osteoarthritis and aging: the results of large-scale cross-sectional study -S. Kana, T. Viik. Annual report of the Estonian Naturalists’ Society 2010 -B. Karmakar, E. Kobyliansky. Finger and palmar dermatoglyphics in Muzeina Bedouins from South Sinai: a qualitative traits -D. Kažoka, J. Vētra. Variations in some anthropometrical parameters of the women with the different iris colour in Latvia -J. Kasmel, T. Kasmel. Seventy years of the Anthropology Section of the Estonian Naturalists’ Society (Part III) -R. Kisenauskaite, D. Paskeviciene. The research of physical condition, physical activity and nutrition of teacher education students -R. Kleina, I. Franckevica, M. Sperga, D. Lutinska. The analysis of undiagnosed malignancies -G. Kolesova, J. Vētra. Sexual dimorphism of pelvic morphology variation in live humans -V. Kulvietis, V. Žalgeviečienė, J. Didžiapetriene,D. Bulotienė, R. Rotomskis. Distribution of nanoparticles in the pregnant rat:the morphologic and spectroscopic study -A. Kuzminienė, S. Šalomskaitė-Davalgienė, I. Balnyte, J. Palubinskienė, A. Valančiūte, V. Ulozas. Evaluation of the chicken embryo choriaollantoic membrane model for laryngeal tumor transplantation -J. Limbo. The tooth size in the end of the Estonian iron age -M. Lintsi, H. Kaarma, M. Aunapuu, A. Arend, R. Aule. Correlation between anthropometrical variables and body surface area -M. Matyk, C. Raschka. Body composition and somatotype of European top roller speed skaters -A. Miskova, M. Pilmane, D. Rezeberga. Immunochistochemical distribution of IGF-1, bFGF and their receptors in decidual embryonic and tubal human pregnancy tissue -E. Mozeika, M. Pilmane, J. Kisis. Distribution of human B-defensin 2,TNF-alpha, IL-1alpha, IL-6 and IL-8 in psoriatic skin -P. Männik, R. T. Kibur, A. Arend, M. Aunapuu. Trophinin and integrin β3 expression in the human endometrium. A pilot study -A. Namm, A. Arend, M. Aunapuu. Bone morphogenetic proteins as regulators of neural tube development -E. Pētersone-Gordina, G. Geshards. Dental disease in a 17th–18th century German community in Jelgava, Latvia -L. Pļaviņa. Assessment of the physical activity level for the staff military personnel -V. Russeva. Alternations of cervical vertebrae in two individuals from the Late Antiquty Necropolis from the “Big Mound” near Cabyle, Bulgaria -S. Skuja, V. Groma, R. Kleina. Chronic alcohol abuse is implicated in the oxidative stress and the changes in the neurotrophic factor receptor expression in the human CNS -R. Stamm, M. Stamm, N. Sorgina, S. Koskel. Training programme to develop young volleyballers’ jumping ability -G. Sumeraga, M. Pilmane. Distribution of neuropeptides in nasal and nasopharyngeal mucosa in patients with the post nasal drip syndrome -N. Szoldatits, B. L. Buda, G. A. Tóth. Body proportion changes of nursing home oligophrenics in western Hungary (1991–2011) -A. Valdovska, M. Pilmane. Relation between serum enzymes and liver histopathology in mink with hepatitis -G. Veldre, T. Kums, E. Salum, J. Eha. Relationship between soldiers’ body height-weight category and changes in their spinal column kyphotic curvature during a long-term military missio

Papers on anthropology XXI

Eelkäija: Tartu Ülikooli toimetised. Antropoloogia-alaseid töid, ISSN 0207-4575http://www.ester.ee/record=b1339521*es

Mediadores de la respuesta celular al desarrollo de obesidad y enfermedad metabólica

Obesity is the pandemic of the 21st century. Overall, about 15% of the world’s adult population is obese, and if the rising trend continues, it is estimated that global prevalence reaches 20% by 2025. Obesity poses a major public health issue due to its elevated risks for adverse health consequences, including several serious chronic diseases, such as insulin resistance (IR), type 2 diabetes (T2D), non-alcoholic fatty liver disease (NAFLD), cardiovascular disease, dyslipidemia, hypertension, stroke, hypercholesterolemia, hypertriglyceridemia, arthritis, asthma, neurodegenerative diseases, and even certain forms of cancer. Many factors and mechanisms have been implicated in obesity pathogenesis, but their trigger is uncertain, and the causal relationship between them and the complications of obesity remains in question. However, there is little doubt that obesity is closely associated with adipose tissue dysfunction, and that this condition leads to the development of metabolic diseases. Adipose tissue is a complex organ with primary roles in energy homeostasis control. Thus, adipose tissue not only acts as a reservoir for energy storage and utilization, but also it senses energy demands and secretes signalling factors to regulate other metabolic tissues. However, in obesity, adipose tissue may become severely dysfunctional and not expand properly to store the energy excess. This induces ectopic fat deposition in other tissues involved in the maintenance of glucose homeostasis, an event commonly defined as “lipotoxicity”. It has been extensively demonstrated that excessive lipid accumulation in ectopic tissues leads to local inflammation and IR. Numerous pathogenic processes have been associated with the unhealthy expansion of the adipose tissue, including inflammation, fibrosis, hypoxia, altered adipokines secretion, mitochondrial dysfunction, hyperinsulinemia, endoplasmic reticulum (ER), and oxidative stress. Dietary and lifestyle, as well as therapeutic interventions, can be adequate to treat obesity and prevent metabolic alterations. Nevertheless, the exact molecular and cellular mechanisms underlying adipose tissue dysfunction and its implications in the development of metabolic disturbances are broadly unknown. Improving our understanding in this field might lead to the development of new approaches and the identification of therapeutic targets for treating obesity. In this Doctoral Thesis, we aimed at identifying potential factors and pathway markers relevant to the loss of adipose tissue function in obesity and their relationship to obesityrelated IR/T2D. To this end, we analysed human adipose tissue samples in combination with both human and murine cell lines, using state-of-the-art methodologies to investigate adipose tissue functionality in obesity and in response to weight loss upon bariatric surgery (BS), and their corresponding mechanisms associated with metabolic alterations or recovery, respectively. Specifically, to reach this general goal, we have carried out two separate studies, which are depicted in detail below. 1. Study 1: Identification of pathogenic markers of metabolic disease in preadipocytes of obese individuals Adipose precursor cells, the preadipocytes, are essential for the maintenance of adipose tissue homeostasis, regeneration, and expansion. Preadipocytes differentiation into adipocytes (i.e., adipogenesis) enables adipocyte turnover and adipose tissue growth, and ensures adipose tissue plasticity to accommodate surplus energy. It has been proposed that the inability for recruiting new adipose cells, together with the functional impairment of hypertrophied adipocytes that occur in obesity, contributes to lipid spillover from the adipose tissue. Thus, increasing adipogenesis appears as a valuable strategy to facilitate healthy adipose tissue expansion and ensure metabolic health. Adipocyte differentiation relies on major changes in gene expression programs regulating mRNA and protein production. An increasing body of evidence shows that mRNA processing and, in particular, alternative splicing, is crucial for genome reprogramming during cell differentiation. However, the splicing components relevant to adipogenesis and the cellular events regulated by alternative splicing during adipocyte differentiation have been scarcely explored, and it is yet to be established whether alternative splicing is modified in human obesity. Another crucial mechanism preserving precursor cell function relates to protein homeostasis (i.e., proteostasis), which maintains the capacity of cells to expand in order to sustain tissue growth and regeneration. Several lines of evidence support an important role for the endoplasmic reticulum (ER) protein quality control system in the regulation of adipogenesis. In fact, the unfolded protein response (UPR) is perturbed in the obese adipose tissue, and it has been proposed to contribute to the pathology of obesity. By contrast, it is still unknown whether the other component of the protein control system, the ER-associated protein degradation (ERAD), which is crucial for protecting cells against the accumulation of misfolded/unfolded proteins and proteotoxicity, is altered in the obese adipose tissue. Here, in order to identify altered molecular pathways that may contribute to metabolic disease in obesity, we set out an iTRAQ-LC-MS/MS proteomic approach for the analysis of subcutaneous (SC) and omental (OM) preadipocytes from obese individuals with normoglycaemia (NG) and T2D. Down-regulation of multiple components of the splicing machinery was observed in SC preadipocytes from obese individuals with insulin resistance (IR) or T2D, as compared to NG obesity. This, together with the observation that adipogenesis can be modulated by regulating the expression levels of a key spliceosome component, PRFP8/PRP8, supports a role for alternative splicing in the development of obesity-associated metabolic complications. In addition, our studies show that not only the UPR is altered in human SC and OM preadipocytes from IR/T2D obese subjects, but also that the ERAD system is hyperactivated. This condition, when mimicked in vitro, prevented adipogenesis. Our results provide novel mechanistic explanations for the impaired adipogenic capacity observed in IR/T2D obesity that relates to both mRNA and ER-proteostasis disturbances. 2. Study 2: Characterization of miRNAs as markers of dysfunctional adipose tissue in type 2 diabetes (T2D) It is increasingly accepted that, besides adipokines, the adipose tissue is a major physiological source of circulating microRNAs (miRNAs). miRNAs, a class of small noncoding RNAs that post-transcriptionally regulate gene expression, have emerged as key molecules for cell function. Anomalous miRNA levels and alterations in their biogénesis machinery have been related to several metabolic diseases, including obesity as well as T2D, and dyslipidemia. miRNAs are also actively secreted into the circulation and have been proposed to act as messengers for intercellular communication. This characteristic has pointed out circulating miRNAs as potential biomarkers for disease and altered circulating levels of numerous miRNAs have been associated with metabolic disorders. In this scenario, in order to design targeted preventive therapeutic strategies, it is critical to identify the early mechanisms that precede disease onset. Recently, the predictive value of a number of miRNAs for the diagnosis of T2D incidence was assessed in the CORonary Diet Intervention with Olive oil and cardiovascular PREVention (CORDIOPREV) study (ClinicalTrial.gov ID: NTC00924937), a prospective study carried out in 1,002 patients with coronary heart disease and high cardiovascular risk. These studies demonstrated that, when combined with HbA1c, a group of nine miRNAs (miR-9, miR-28-3p, miR-29a, miR-30a-5p, miR- 103, miR-126, miR-150, miR-223-3p, and miR-375), provided a higher predictive value in T2D diagnosis than clinical parameters. The relationship between baseline levels of these miRNAs with markers of beta-cell function and systemic and peripheral IR was also investigated. However, their potential association with adipose tissue deregulation was not analyzed. This is of interest as adipose tissue dysfunction has been proposed as a major contributing factor for the development of T2D, and adipose tissue miRNA expression profile is altered in obesity and T2D. In this line, several cross-sectional human studies have shown that the obesity-related expression pattern of specific circulating miRNAs reflects their miRNA adipose tissue expression profiles, supporting a role for circulating miRNAs as adipose tissue biomarkers. However, the relationship between circulating miRNAs and adipose tissue functional state is not fully understood, yet it might be useful to identify adipose tissue-related metabolic complications. In this scenario, we aimed at analyzing the relationship between previously established predictive miRNAs for T2D onset in the CORDIOPREV cohort (Jiménez- Lucena et al., 2018) and the loss of adipose tissue insulin sensitivity. Once established that circulating miR-223-3p was dysregulated in relation to adipose tissue function, we performed functional analysis to elucidate both the potential of preadipocytes and adipocytes as miR-223-3p secreting cells, and the consequences of miR-223-3p dysregulation on adipocyte biology. Our results indicate that miR-223-3p secretion by preadipocytes and adipocytes is prevented under inflammatory conditions, and that its pathogenic accumulation leads to alterations in both glucose and lipid metabolism in these cells. These observations could answer to the lower circulating levels on this miRNA found in those patients who are going to develop T2D, as well as explain its underlying role as a potential predictor of adipose dysfunction related to T2D development. We have also demonstrated that the changes that occur in the circulating milieu after BS-induced weight-loss modulate miR-223-3p expression by adipocytes. The relevance of these observations was further supported by our functional studies in adipocytes exposed to pre- and post-BS serum, where the post-BS serum improved the insulininduced glucose uptake in adipocytes, in comparison with those cells exposed to pre- BS. These findings further support the notion that inflammatory mediators present in either the serum, or locally in the adipose tissue, may be responsible for the alterations in miR-223-3p regulation. In sum, when viewed together, our studies indicate that impaired splicing and disturbed ER-proteostasis are components of the pathogenic molecular fingerprint of preadipocytes that could be targeted to prevent and/or improve adipose tissue dysfunction in obesity and its related metabolic disorders. Likewise, our data support the notion that miR-223-3p may have a role as a potential predictor of adipose dysfunction related to T2D development, thus unveiling a novel molecular target that may be helpful to design novel therapeutic strategies to prevent T2D.La obesidad es la pandemia del siglo XXI. En conjunto, alrededor del 15% de la población adulta mundial es obesa. Si la tendencia al alza continúa, se estima que la prevalencia mundial alcanzará el 20% para el 2025. Así, la obesidad plantea un problema importante de salud pública debido a sus elevados riesgos para la salud, entre ellas varias enfermedades crónicas graves, como resistencia a insulina (IR), diabetes tipo 2 (T2D), enfermedad de hígado graso no alcohólico (NAFLD), enfermedad cardiovascular, dislipidemia, hipertensión, accidente cerebrovascular, hipercolesterolemia, hipertrigliceridemia, artritis, asma, enfermedades neurodegenerativas e incluso ciertas formas de cáncer. Diversos factores y mecanismos han sido implicados en la patogénesis de la obesidad, pero su desencadenante es incierto. Además, la relación causal entre ellos y las complicaciones de la obesidad permanece sin resolver. Sin embargo, hay pocas dudas de que la obesidad está estrechamente asociada con la disfunción del tejido adiposo, y que esta condición conduce al desarrollo de enfermedades metabólicas. El tejido adiposo es un órgano complejo con funciones principales en el control de la homeostasis energética. Por lo tanto, el tejido adiposo no solo actúa como un depósito para el almacenamiento y la utilización de energía, sino que también detecta las demandas de energía, y secreta factores de señalización para regular otros tejidos metabólicos. Sin embargo, en obesidad, el tejido adiposo puede volverse gravemente disfuncional, y no expandirse adecuadamente para almacenar el exceso de energía. Esto induce el depósito ectópico de grasa en otros tejidos involucrados en el mantenimiento de la homeostasis de la glucosa, un evento comúnmente definido como "lipotoxicidad". Se ha demostrado ampliamente que la acumulación excesiva de lípidos en tejidos ectópicos provoca inflamación local e IR. Numerosos procesos patogénicos se han asociado con la expansión no saludable del tejido adiposo, que incluyen: inflamación, fibrosis, hipoxia, secreción alterada de adipoquinas, disfunción mitocondrial, hiperinsulinemia, y estrés de retículo endoplásmico (ER) y oxidativo. Las intervenciones dietéticas y de estilo de vida, así como las terapéuticas, pueden ser adecuadas para tratar la obesidad y prevenir alteraciones metabólicas. Sin embargo, los mecanismos moleculares y celulares exactos que subyacen a la disfunción del tejido adiposo, y sus implicaciones en el desarrollo de alteraciones metabólicas son ampliamente desconocidos. Mejorar nuestra comprensión en este campo podría conducir al desarrollo de nuevos enfoques y a la identificación de dianas terapéuticas para tratar la obesidad. En esta Tesis Doctoral, nuestro objetivo fue identificar los posibles factores y marcadores de las vías relevantes para la pérdida de la función del tejido adiposo en obesidad, y su relación con IR/T2D asociadas. Para ello, analizamos muestras de tejido adiposo humano en combinación con líneas celulares, tanto humanas como murinas, utilizando metodologías de vanguardia para investigar la funcionalidad del tejido adiposo en obesidad, y su respuesta a la pérdida de peso tras cirugía bariátrica (BS). Así como, sus correspondientes mecanismos asociados con alteraciones metabólicas o de recuperación, respectivamente. Específicamente, para alcanzar este objetivo general, hemos llevado a cabo dos estudios separados, que se describen en detalle a continuación. 1. Estudio 1: Identificación de marcadores patogénicos de enfermedad metabólica en preadipocitos de individuos obesos Las células adiposas precursoras, los preadipocitos, son esenciales para el mantenimiento de la homeostasis, regeneración y expansión del tejido adiposo. La diferenciación de los preadipocitos en adipocitos (adipogénesis) permite el recambio de adipocitos y el crecimiento del tejido adiposo, y asegura la plasticidad de este tejido para acomodar el exceso de energía. Se ha propuesto que la incapacidad para reclutar nuevas células adiposas, junto con el deterioro funcional de los adipocitos hipertróficos que ocurre en obesidad, contribuye al derrame de lípidos del tejido adiposo. Por lo tanto, el aumento de la adipogénesis aparece como una estrategia valiosa para facilitar la expansión saludable del tejido adiposo y salvaguardar la salud metabólica. La diferenciación de adipocitos se basa en cambios importantes en los programas de expresión génica que regulan la producción de ARNm y proteínas. Un creciente número de evidencias muestra que el procesamiento del ARNm y, en particular, el empalme alternativo, es crucial para la reprogramación del genoma durante la diferenciación celular. Sin embargo, los componentes de empalme relevantes para la adipogénesis y los eventos celulares regulados por este empalme alternativo durante la diferenciación de adipocitos apenas se han explorado. Así mismo, aún no se ha establecido si el empalme alternativo se modifica en la obesidad humana. Otro mecanismo crucial que preserva la función de las células precursoras se relaciona con la homeostasis de las proteínas (proteostasis), que mantiene la capacidad de las células de expandirse para mantener el crecimiento y la regeneración de los tejidos. Varias líneas de evidencia apoyan un papel importante para el sistema de control de calidad proteico del retículo endoplásmico (ER) en la regulación de la adipogénesis. De hecho, la respuesta a proteínas mal plegadas (UPR) se perturba en el tejido adiposo obeso, y se ha propuesto que contribuye a la patología de la obesidad. Por el contrario, todavía se desconoce si otro componente del sistema de control de proteínas, la degradación de proteínas asociada a ER (ERAD), que es crucial para proteger las células contra la acumulación de proteínas mal plegadas/desplegadas y la proteotoxicidad, está alterado en el tejido adiposo obeso. Aquí, con el fin de identificar vías moleculares alteradas que pueden contribuir a la enfermedad metabólica en obesidad, realizamos un estudio proteómico de iTRAQ-LCMS/ MS para el análisis de preadipocitos subcutáneos (SC) y omentales (OM) de individuos obesos con normoglucemia (NG) y T2D. Se observó una disminución de múltiples componentes de la maquinaria de empalme en preadipocitos SC de individuos obesos con IR o T2D, en comparación con los obesos NG. Esto, junto con la observación de que la adipogénesis puede modularse regulando los niveles de expresión de un componente clave del espliceosoma, PRPF8/PRP8, respalda el papel de este proceso en el desarrollo de complicaciones metabólicas asociadas a la obesidad. Además, nuestros estudios muestran que no solo el sistema UPR está alterado en preadipocitos humanos SC y OM de sujetos obesos con IR/T2D, sino que también existe una hiperactivación del sistema ERAD. Esta condición, cuando se imita in vitro, previene la adipogénesis. Nuestros resultados proporcionan novedosas explicaciones mecanicistas para la capacidad adipogénica deteriorada observada en los obesos IR/T2D que se relaciona tanto con la alteración del procesamiento de ARNm como con la de la proteostasis del ER. 2. Estudio 2: Caracterización de miRNAs como marcadores de tejido adiposo disfuncional en la diabetes tipo 2 (T2D) Cada vez se acepta más que, además de las adipoquinas, el tejido adiposo es una importante fuente fisiológica de microRNAs (miRNAs) circulantes. Los miRNAs, una clase de pequeños ARN no codificantes que regulan la expresión génica posttranscripcionalmente, han surgido como moléculas clave para la función celular. Los niveles de miRNAs anómalos y las alteraciones en su maquinaria de biogénesis se han relacionado con varias enfermedades metabólicas, incluida la obesidad, así como la T2D y la dislipidemia. Los miRNAs también se secretan activamente a la circulación y se ha propuesto que actúen como mensajeros de la comunicación intercelular. Esta característica ha señalado a los miRNAs circulantes como posibles biomarcadores de enfermedades, y los niveles circulantes alterados de numerosos miRNAs se han asociado con trastornos metabólicos. En este escenario, para diseñar estrategias terapéuticas preventivas específicas, es fundamental identificar los mecanismos tempranos que preceden al inicio de la enfermedad. Recientemente, en el estudio CORonary Diet Intervention with Olive oil and cardiovascular PREVention (CORDIOPREV, ClinicalTrial.gov ID: NTC00924937), se evaluó el valor predictivo de varios miRNAs para el diagnóstico de incidencia de T2D, un estudio prospectivo realizado en 1,002 pacientes con enfermedad coronaria y alto riesgo cardiovascular. Estos estudios demostraron que, cuando se combina con HbA1c, un grupo de nueve miRNAs (miR-9, miR-28-3p, miR-29a, miR-30a-5p, miR-103, miR- 126, miR-150, miR-223-3p, y miR-375), proporcionaron un valor predictivo más alto en el diagnóstico de T2D que los parámetros clínicos. También se investigó la relación entre los niveles basales de estos miRNAs con marcadores de la función de las células beta y la IR sistémica y periférica. Sin embargo, su posible asociación con la desregulación del tejido adiposo no se analizó. Esto es de interés ya que se ha propuesto que la disfunción del tejido adiposo es un factor importante que contribuye al desarrollo de T2D, y el perfil de expresión de miRNAs del tejido adiposo está alterado en obesidad y T2D. En esta línea, varios estudios transversales en humanos han demostrado que el patrón de expresión de específicos miRNAs circulantes, relacionados con obesidad, refleja los perfiles de expresión de estos miRNAs en el tejido adiposo, lo que respalda el papel de los miRNAs circulantes como biomarcadores del tejido adiposo. Sin embargo, la relación entre los miRNAs circulantes y el estado funcional del tejido adiposo no se comprende completamente, lo cual podría ser útil para identificar las complicaciones metabólicas relacionadas con el tejido adiposo. En este escenario, nuestro objetivo fue analizar la relación entre los miRNAs predictivos para el inicio de T2D, previamente establecidos en la cohorte CORDIOPREV (Jiménez-Lucena et al., 2018), y la pérdida de sensibilidad a insulina del tejido adiposo. Una vez establecido que el miR-223-3p circulante estaba desregulado con relación a la función del tejido adiposo, realizamos un análisis funcional para dilucidar tanto el potencial de los preadipocitos y los adipocitos como células secretoras de miR-223-3p, como las consecuencias de la desregulación de este miRNA sobre la biología de los adipocitos. Así, nuestros resultados indican que la secreción de miR-223-3p por preadipocitos y adipocitos es inhibida en condiciones inflamatorias, y que su acumulación patológica provoca alteraciones tanto en el metabolismo glucídico como lip

Inimese endomeetriumi normaalne ja patoloogiline profiil proteoomika vaatevinklist

Väitekirja elektrooniline versioon ei sisalda publikatsiooneDNA ja RNA-põhiste meetodite kasutamine on reproduktiivmeditsiini arengut märkimisväärselt mõjutanud, suurendades arusaamist haiguste pärilikest tagamaadest ja tuues kliinilisse praktikasse uusi diagnostilisi meetodeid. Geenide peamiseks funktsionaalseks väljundiks on aga valgud ning nende uurimine on ainus viis haiguslikest protsessidest vahetu ja detailsema pildi saamiseks. Seda enam, et esmase geeniekspressiooni seos valkude tasemetega rakkudes, kudedes ja kehavedelikes on tihtilugu piiratud. Valkude uurimiseks kasutatav massispektromeetrial põhinev proteoomika on kiirelt arenev teadusharu, mis võimaldab bioloogilisest materjalist, st. nii kudedest, rakkudest kui kehavedelikest kõrge tundlikkusega määrata nende valgulist koostist ehk proteoomi. Antud uurimistöö eesmärgiks oligi rakendada kaasaegseid massispektromeetria-põhiseid proteoomika meetodeid, et leida täpsemaid vastuseid naiste reproduktiivtervisega seotud probleemide molekulaarsete mehhanismide selgitamiseks ja välja pakkuda uusi markerikandidaate diagnostikaks. Täpsemalt keskendus meie töö endometrioosile ja kehavälise viljastamise (IVF) efektiivsusega seotud biomarkerite otsingule. Meie tööst selgus, et endometrioosi korral on haiguskolde rakkudes toimunud mitmeid muutusi, mis sarnanevad vähi- ja tüvirakkudes kirjeldatud ainevahetuslike kohastumustega. Meie tulemused näitavad, et endometrioosikolde rakud kasutavad sõltumata hapniku juuresolust suurenenud määral anaeroobset ainevahetust. Samuti on kolderakkudes muutunud valkude avaldumine, mis on seotud rakkude levimise ja ellujäämisega. Meie tulemused pakuvad küll välja potentsiaalseid sihtmärk-valke haiguskulu mõjutamiseks, kuid rõhutavad ka vajadust edasisteks uuringuteks suuremates kohortides. Töö teises osas uurisime emakaõõne sekreedi valke, et leida biomarkereid kehavälise viljastamise optimeerimiseks. Selle töö tulemusena leidsime, et emakaõõne sekreet sisaldab valke, mida on võimalik rakendada embrüosiirdamise efektiivsemaks ajastamiseks. Samuti viitavad meie tulemused, et naistel, kellel on IVF korduvalt ebaõnnestunud, esineb häireid emaka limaskesta vastuvõtlikkuses embrüotele ehk retseptiivsuses. Leitud valguliste biomarkerite paneeli on võimalik arendada diagnostiliseks testiks, mis tõstaks IVFi tõhusust ja aitaks paremini tuvastada retseptiivsushäirega naisi.The impact of genomics and transcriptomics methods for reproductive medicine has been substantial by increasing the understanding behind heritable factors of diseases and by enabling new avenues for diagnostic testing. Nevertheless, the main functional output of genes are proteins, and, studying proteins is essential for providing further detail into the understanding of pathogenetic mechanisms. Furthermore, the correlation between early gene expression and the levels of proteins in cells, tissues and bodily fluids has been shown to be limited. Mass spectrometry-based proteomics is a progressively developing field that has finally reached to a point where measurement of proteomes or the full protein complement is now becoming feasible. The main objective of the current work was to apply modern proteomics methods to enhance the understanding behind reproductive diseases and to propose new biomarkers for diagnostics. More specifically, our work focused on endometriosis and the search for markers that could improve the in vitro fertilization (IVF) process. By analyzing cells from the uterine lining and endometriosis lesions, we found evidence for altered energy metabolism in endometriotic stromal cells and these changes are akin to changes previously seen in tumor and stem cells. In addition, lesion stromal cells appear to have changes in the expression of proteins that are associated with increased invasiveness and survival of cells. Our results propose novel candidates for future studies to see whether targeting these proteins could affect the course of the illness. In the second part of our work, we analyzed secreted proteins from uterine cells to find biomarkers for improving the success rates of IVF. As a result of our study, we found that the uterine secretome contains proteins that reflect the embryo receptivity status of the endometrium. Our results also underlined that women with recurrent implantation failure in IVF have altered uterine secretomes. The proposed biomarker panel of our study can be developed into a diagnostic test that would increase the effectiveness of the IVF procedure and would help to screen for women with pathological uterine receptivity.https://www.ester.ee/record=b534053