Bone Mineral Accrual in Physically Active Girls. With Special Reference to Reduction in Physical Activity Level and Use of Oral Contraceptives

Adolescence is an important time for acquiring high peak bone mass. Physical activity is known to be beneficial to bone development. The effect of estrogen-progestin contraceptives (EPC) is still controversial. Altogether 142 (52 gymnasts, 46 runners, and 42 controls) adolescent women participated in this study, which is based on two 7-year (n =142), one 6-year (n =140) and one 4-year (n =122) follow-ups. Information on physical activity, menstrual history, sexual maturation, nutrition, living habits and health status was obtained through questionnaires and interviews. The bone mineral density (BMD) and content (BMC) of lumbar spine (LS) and femoral neck (FN) were measured by dual- energy X-ray absoptiometry. Calcaneal sonographic measurements were also made. The physical activity of the athletes participating in this study decreased after 3-year follow-up. High-impact exercise was beneficial to bones. LS and FN BMC was higher in gymnasts than in controls during the follow-up. Reduction in physical activity had negative effects on bone mass. LS and FN BMC increased less in the group having reduced their physical activity more than 50%, compared with those continuing at the previous level (1.69 g, p=0.021; 0.14 g, p=0.015, respectively). The amount of physical activity was the only significant parameter accounting for the calcaneal sonography measurements at 6-year follow-up (11.3%) and reduced activity level was associated with lower sonographic values. Long-term low-dose EPC use seemed to prevent normal bone mass acquisition. There was a significant trend towards a smaller increase in LS and FN BMC among long-term EPC users. In conclusion, this study confirms that high-impact exercise is beneficial to bones and that the benefits are partly maintained even after a clear reduction in training level at least for 4 years. Continued exercise is needed to retain all acquired benefits. The bone mass gained and maintained can possibly be maximized in adolescence by implementing high-impact exercise for youngsters. The peak bone mass of the young women participating in the study may be reached before the age of 20. Use of low-dose EPCs seems to suppress normal bone mass acquisition.

Regulation of Spermatogenesis: Differentiation of GFP-labeled Stem Cells and the Function of Cytoplasmic Bridges

During spermatogenesis, different genes are expressed in a strictly coordinated fashion providing an excellent model to study cell differentiation. Recent identification of testis specific genes and the development of green fluorescence protein (GFP) transgene technology and an in vivo system for studying the differentiation of transplanted male germ cells in infertile testis has opened new possibilities for studying the male germ cell differentiation at molecular level. We have employed these techniques in combination with transillumination based stage recognition (Parvinen and Vanha-Perttula, 1972) and squash preparation techniques (Parvinen and Hecht, 1981) to study the regulation of male germ cell differentiation. By using transgenic mice expressing enhanced-(E)GFP as a marker we have studied the expression and hormonal regulation of beta-actin and acrosin proteins in the developmentally different living male germ cells. Beta-actin was demonstrated in all male germ cells, whereas acrosin was expressed only in late meiotic and in postmeiotic cells. Follicle stimulating hormone stimulated b-actin-EGFP expression at stages I-VI and enhanced the formation of microtubules in spermatids and this way reduced the size of the acrosomic system. When EGFP expressing spermatogonial stem cells were transplanted into infertile mouse testis differentiation and the synchronized development of male germ cells could be observed during six months observation time. Each colony developed independently and maintained typical stage-dependent cell associations. Furthermore, if more than two colonies were fused, each of them was adjusted to one stage and synchronized. By studying living spermatids we were able to demonstrate novel functions for Golgi complex and chromatoid body in material sharing between neighbor spermatids. Immunosytochemical analyses revealed a transport of haploid cell specific proteins in spermatids (TRA54 and Shippo1) and through the intercellular bridges (TRA54). Cytoskeleton inhibitor (nocodazole) demonstrated the importance of microtubules in material sharing between spermatids and in preserving the integrity of the chromatoid body. Golgi complex inhibitor, brefeldin A, revealed the great importance of Golgi complex i) in acrosomic system formation ii) TRA54 translation and in iii) granule trafficking between spermatids.

Novel Roles of HCG/LH Signalling in the Mouse Mammary Gland and its Tumorigenesis.

Breast cancer is the most common cancer in women, and its development is intimately related to hormonal factors, but how hormones affect breast physiology and tumorigenesis is not sufficiently known. Pregnancy elicits long-term protection from breast cancer, but during the first ten years after pregnancy, breast cancer risk is increased. In previous studies, there has been conflicting data on the role of human chorionic gonadotropin (HCG) and the functionality of its receptor in extragonadal tissues. The aim of this study was to elucidate the role of chronically elevated HCG in mouse physiology. We have created a transgenic (TG) mouse model that overexpresses HCG. HCG is similar to lutenizing hormone (LH), but is secreted almost solely by the placenta during pregnancy. HCG and LH both bind to the LH receptor (LHR). In the current study, mammary gland tumors were observed in HCG TG mice. We elucidated the role of HCG in mammary gland signalling and the effects of LHR mediated signalling in mouse mammary gland gene expression. We also studied the effects of HCG in human breast epithelial cell cultures. Several endocrine disturbances were observed in HCGβ TG female mice, resulting in precocious puberty, infertility, obesity and pituitary and mammary gland tumors. The histology of the mammary gland tumors of HCGβ TG females resembled those observed in mouse models with activated Wnt/β-catenin signalling pathway. Wnts are involved in stem cell regulation and tumorigenesis, and are hormonally regulated in the mammary gland. We observed activated β-catenin signalling and elevated expression of Wnt5b and Wnt7b in TG tumors and mammary glands. Furthermore, we discovered that HCG directly regulates the expression of Wnt5b and Wnt7b in the mouse mammary gland. Pharmacological treatment with HCG also caused upregulation of several Wnt-pathway target genes in ovariectomized wild type (WT) mice in the presence of physiological concentrations of estradiol and progesterone. In addition, differential expression of several metabolic genes was observed, suggesting that HCG affects adipocyte function or glucose metabolism. When WT mice were transplanted with LHR deficient or wild type WT mammary epithelium, differential expression of several genes affecting the Wnt-signalling pathway was observed in microarray analysis. Diminished expression of several genes associated with LHR function in other tissues, such as the ovary, was observed in mammary glands deficient of epithelial LHR. In cultured human mammary epithelial cells HCG upregulated the expression of WNT5B, WNT7B similar to mouse, suggesting that the observations found are relevant in human physiology. These studies suggest that HCG/LHR signalling affects gene expression in non-gonadal tissues, and that Wnt-signalling is regulated by HCG/LH in human and mouse mammary glands.

Venäläistä rulettia – Nuorten perhesuunnittelun esteet ja tarpeet Pietarissa ja Riiassa

Latviassa ja Venäjällä heikko demografinen tilanne on lisännyt keskustelua seksuaali- ja kansalaisoikeuksista sekä valtion merkityksestä niiden toteuttamisessa. Väestöpolitiikassa näkyy yhteiskunnallinen tilanne sekä sen kyky ja mahdollisuudet huomioida nuorten perhesuunnitteluun liittyviä tarpeita. Huomion kohteena on myös palvelujärjestelmä ja siinä toteutuva yksilöllinen taso. Tutkin perhesuunnittelua kokonaisvaltaisesti, mikä ilmenee erilaisten polkuriippuvuuksien tarkasteluina. Mielenkiintoiseksi kohteen tekee entisten sosialististen valtioiden erityislaatuisen murroksen läpikäyminen. Perhesuunnittelu on osa seksuaali- ja ihmisoikeuksia. Siihen kuuluu seksuaaliterveydestä tiedottaminen ja ohjaus sekä siihen liittyvät toimenpiteet. Perhesuunnittelu sisältää niin sosiaaliset, kasvatukselliset kuin lääketieteellisetkin näkökulmat. Siihen liitetään myös moraalisia ja taloudellisia näkemyksiä, sekä politiikkaa ja henkilökohtaisuutta. Tutkimustehtäväni on nuorten perhesuunnittelun esteet ja tarpeet Riiassa ja Pietarissa. Sovellan metodologisena lähestymistapana sosiaalihistoriallista ja etnografista tutkimusotetta, jolloin perinteen vaikutusten, kulttuuristen rakenteiden ja yksilöllisten toimintatapojen ymmärtäminen mahdollistuu. Aineisto koostuu viranomaishaastatteluista sosialismin kaudella toteutuneesta sekä nykyisestä perhesuunnittelusta ja nuorten haastatteluista Pietarissa ja Riiassa. Perehdyn myös lisääntymisterveyteen liittyviin sanomalehtiartikkeleihin sekä Latviassa että Venäjällä. Lisäksi havainnoin seksuaalineuvontatilanteita Pietarissa. Sekä sosialismin aikana että nykyään painottuvat väestönkasvun odotukset, joita tuetaan sukupuoli- ja moraalikasvatuksen avulla. Perhesuunnittelun esteiden analyysin tuloksissa on nähtävissä sosiaalisten olosuhteiden ja rakenteiden merkitys. Jälkisosialistisesta riskiyhteiskunnasta puuttuvat yhteiskuntaan integroitumisen mahdollistavat instituutiot. Tämä ilmenee yhteiskunnallisena vastuuttomuutena. Universaalit palvelut ovat suurelta osin peräisin sosialismin ajalta. Kaupalliset yritykset tekevät lähinnä teknisiä, yksittäisiä ehkäisyvälineisiin liittyviä interventioita nuorten elämään. Teini-ikäiset jätetään oman harkintansa varaan. Latviassa perhesuunnitteluun liittyvä ennalta ehkäisevä toiminta on huomioitu uudessa lisääntymisterveyslaissa, mutta sen heikko toteutus tuo sen lähelle neuvostoaikaista kulttuurikonventiota, jossa määrällisillä suoritteilla ja sekundaaripreventiolla on arvoa. Venäjällä voimavarat kohdentuvat jälkihuollon palveluihin. Foucaultin valta-analyysi kertoo strategiasta, jossa ihminen sisäistää vallankäytön osaksi omaa ajatteluaan ja toimintaansa. Tässä näkökulmassa yksilöllinen, yhteiskunnallinen ja historiallinen ulottuvuus limittyvät toisiinsa. Vallan muodot ovat yhteiskunnan rakenteissa. Väestöpolitiikan avulla tuotetaan tietoa ja muovataan hallitsemisen kohteita. Osa haastatelluista nuorista omaksui yhteiskunnan passiivisuuden osaksi omaa toimintaansa. He eivät tiedostaneet palvelujärjestelmän puutteita eivätkä kyseenalaistaneet sen oikeutusta. Myös lehtiartikkelit ja viranomaishaastattelut tukivat osittain seksuaalikasvatuksen sivuuttamista. Silloin hyväksyttävänä totuutena pidettiin väestöpolitiikkaa, jossa on annettu tilaa populistisille suuntauksille. Sekä Latviassa että Venäjällä ilmaistiin tavoitteita väestön kasvusta. Kansainvälisten seksuaalioikeuksien maihinnousu ei ole poistanut ääri uskonnollisten ja osittain kansalaisjärjestöjen puitteissa tapahtuvaa perhesuunnittelun vastustusta. Näissä suuntauksissa vaikutetaan ihmisten moraaliin ja vastustetaan yhteiskunnan interventioita kieltämällä seksuaalikasvatus. Sosialismin aikana sosiaali- ja terveydenhuoltoa toteutettiin totalitaristisessa, suljetussa yhteiskunnassa tiukasti rajatuilla voimavaroilla. Tuolta ajalta peräisin oleva viranomaisten yleinen medikalisoitunut ja tekninen lähestymistapa estää osittain nuoren valtaistumista. Vieläkin heikko tiedottaminen ja puutteellinen koulutus vaikuttavat ammattilaisten keskuudessa. Yksi haitallisista perhesuunnitteluun kuuluvista uskomuksista oli hormonaalisen ehkäisyn aiheuttamat ongelmat. Lisäksi pelättiin heikkotasoisesti tehtyjen aborttien aiheuttavan hedelmättömyyttä. Uskomukset ovat eläneet vieläkin, kun osa asiantuntijoista kannusti ensimmäisen raskauden päättymistä synnytykseen. Näillä käsityksillä on ollut taipumus siirtyä sukupolvelta toiselle myös kansalaisten keskuudessa. Sukuyhteisöjen ja verkostojen oleellinen merkitys teini-ikäisen arjessa ja selviytymisessä painotti perinteistä sosialisaatiota. Uskomusten lisäksi nuorten heikko taloudellinen tilanne on ohjannut heitä ehkäisyssä luonnonmenetelmien käyttöön. Neuvostoaikainen seksuaalikielteisyyden perinne on vaikuttanut myös sukupolvien kyvyttömyyteen keskustella aiheesta. Yleisen ennaltaehkäisevän neuvonnan puuttuminen on suunnannut suurta osaa nuoria mallioppimiseen, jossa esimerkit ovat nousseet satunnaisista löydöistä. Toisaalta työntekijöiden neuvokkuus ja aloitteellisuus erilaisissa ohjaamistilanteissa, toimintatavoissa ja instituutioiden perustamisissa kuvaavat eettistä vastuunottoa ja paneutumista nuorten perhesuunnittelun toteutumiseen sekä sen mahdollisuuksiin. Perhesuunnitteluun liittyvän tiedonsaannin parantamiseksi viranomaiset ovat aloittaneet nuorten vertaisryhmiä. Lisäksi yhtenä uutena virallisena lähestymistapana oli neuvostoaikana kielletty psykoterapeuttisen suuntauksen avoin käyttöönotto. Myös nuorille suunnattuja palveluja niin lainsäädännön kuin instituutioiden tasolla on saatu aikaan. Nämä myönteiset tapaukset jäivät kuitenkin yksittäisiksi, osittain joidenkin työntekijöiden omakohtaisen sitoutumisen varaan toimiviksi. Tutkimuksessa nousee esiin yhteiskunnan vastuu erityisesti haavoittuvien ryhmien kohdalla. Nuoret ovat tiedoiltaan ja kokemuksiltaan heikossa asemassa. Sosiaalipoliittisesta näkökulmasta julkisilla ja ennaltaehkäisevillä palveluilla on keskeinen merkitys etenkin niiden kohdalla, jotka tarvitsevat runsaasti palveluja. Usein tähän ryhmään kuuluvat ovat passiivisia tiedon hakijoita. Tutkimus tuo esille perhesuunnittelun palvelujärjestelmään ja sen kohderyhmään kuuluvan moniulotteisuuden, jossa toisiinsa nivoutuvat kulttuuriset myytit, uskomukset, tabut, toimintatavat ja tietämys. Näiden polkujen avaaminen tässä tutkimuksessa edistää perhesuunnittelun esteiden tunnistamista ja niihin soveltuvien ratkaisukeinojen löytymistä.

The Effects of Estradiol, Androgens and the Selective Estrogen Modulators: Ospemifene, Raloxifene and Tamoxifen on Explant Cultures of Human Breast Tissue

The impact of menopausal hormone therapy (MHT) on increasing the risk for breast cancer (BC) remains controversial. To understand MHT-elicited cellular breast effects and the potential risks, included with using this therapy, a further investigation into this controversy is the subject of this thesis. In this thesis, to study the effects of estrogen, progestin, androgens and selective estrogen receptor modulators (SERMs), a modified tissue explant culture system was used. The different types of human breast tissues (HBTs) used in this study were normal HBTs, obtained from reduction mammoplasties of premenopausal women (prem-HBTs) or postmenopausal (postm-HBTs) women and peritumoral HBTs (peritum-HBTs) which were obtained from surgeries on postmenopausal BC patients. The explants were cultured up to three weeks in the presence or absence of estradiol (E2), medroxyprogesterone acetate (MPA), testosterone (T), dihydrotestosterone (DHT) and SERMs - ospemifene (OSP), raloxifene (RAL) and tamoxifen (TAM). The cultured HBTs maintained morphological integrity and responded to hormonal treatment in vitro. E2, MPA or E2/MPA increased proliferative activity and was associated with increased cyclin-D1 and caused changes in the cell cycle inhibitors p21 and p27, whereas the androgens T and DHT inhibited proliferation and increased apoptosis in HBT epithelia and opposed E2-stimulated proliferation and cell survival. The postm-HBTs were more sensitive to E2 than prem-HBTs. The effects of OSP, RAL and TAM on HBT epithelium were antiproliferative. E2, androgens and SERMs were associated with marked changes in the proportions of epithelial cells expressing steroid hormone receptors: E2 increased ERα expressing cells and decreased androgen receptor (AR) positive cells, whereas T and DHT had opposite effects. The OSP, RAL and TAM, also decreased a proportion of ERα positive cells in HBT epithelium. At 100 nM, these compounds maintained the relative number of AR positive cells, present at control level, which may partly explain proliferative inhibition. In conclusion, the proliferative activity of E2, in the epithelium of postm-HBTs, is opposed by T and DHT, which suggests that the inclusion of androgens in MHT may decrease the risk for developing BC.

Multiple sclerosis and pregnancy: clinical and immunological aspects

Background: Multiple Sclerosis (MS) is an autoimmune disease of the central nervous system that affects most commonly young women in their childbearing age. Previous studies have shown that MS relapse rate usually reduces during pregnancy and increases again after delivery. Patients with MS and their treating physicians are interested to know more about the risks the disease can cause to pregnancy and how pregnancy affects the disease. The reasons for increased relapse rate after delivery are not entirely clear, but loss of pregnancy related immune tolerance and changes in the hormonal status at the time of delivery seem to be of relevance. Aims and methods: The aims of this study were to follow the natural course of MS during and after pregnancy, evaluate pregnancy related risks among MS patients, follow the inflammatory response of MS patients during and after pregnancy and clarify the risk of relevant co-morbidities known to affect other autoimmune diseases after pregnancy and compare these results to healthy controls. This study was a part of a prospective nation-wide follow-up study of 60 Finnish MS patients. All eligible MS patients were enrolled in the study during the years 2003-2005. A prospective followup continued from early pregnancy until six months postpartum. MS relapses, EDSS scores and obstetric details were recorded. Blood samples were obtained from the patients at early, middle, and late pregnancy, after delivery and one month, three months and six months postpartum. Results: MS patients were no more likely to experience pregnancy or delivery complications than the Finnish mothers in general. The need of instrumental assistance, however, was higher among mothers with MS. Disease activity followed the course seen in previous studies. The majority of mothers (90.2%) breastfed their babies. Contrary to previous results, breastfeeding did not protect MS patients from disease worsening after delivery in present study. Mothers with active pre-pregnancy disease chose to breastfeed less frequently and started medication instead. MS patients presented with higher prevalence of elevated thyroid autoantibodies postpartum than healthy controls, but the rate of thyroid hormonal dysfunction was similar as that of healthy controls. The mode of delivery nor the higher rate of tissue damage assessed with C-reactive protein concentration were not predictive of postpartum relapses. The prevalence of gestational diabetes was slightly higher among mothers with MS compared to Finnish mothers in general, but postpartum depression was observed in similar rates. MS patients presented with significantly lower serum concentrations of vitamin D during pregnancy and postpartum than healthy controls. Conclusions: Childbearing can be regarded as safe for mothers with MS as it is for healthy mothers in general. Breastfeeding can be recommended, but it should be done only after careful evaluation of the individual risk for postpartum disease activation. Considering MS patients tend to develop thyroid antibody positivity after delivery more often than healthy controls and that certain treatments can predispose MS patients to thyroid hormonal dysfunction, we recommend MS mothers to be screened for thyroid abnormalities during pregnancy and after delivery. Increased risk for gestational diabetes should be kept in mind when following MS mothers and glucose tolerance test in early pregnancy should be considered. Adequate vitamin D supplementation is essential for MS mothers also during pregnancy and postpartum period.

Oxidized LDL lipids as a risk factor for atherosclerosis

University of Turku, Faculty of Medicine, Department of Clinical Medicine, Department of Physical Activity and Health, Paavo Nurmi Centre, Doctoral Programme of Clinical Investigation, University of Turku, Turku, Finland. Annales Universitatis Turkuensis. Medica – Odontologica, Turku, Finland, 2014. Background: Atherosclerosis progression spans an entire lifetime and has a wide pool of risk factors. Oxidized LDL (oxLDL) is a crucial element in the progression of atherosclerosis. As a rather new member in the atherosclerosis risk factor family, its interaction with the traditional pro-atherogenic contributors that occur at different ages is poorly known. Aims: The aim of this study was to investigate oxLDL and its relation to major contributing risk factors in estimating atherosclerosis risk in data consisting mostly of adult men. The study subjects of this study consisted of four different sets of data, one of which contained also women. The age range of participants was 18-100 years and totaled 2337 participants (of whom 69% were men). Data on anthropometric and hormonal parameters, laboratory measures and medical records were assessed during 1998-2009. Results: Obesity was paralleled with high concentrations of oxLDL, which consequentially was reduced by weight reduction. Importantly, successful weight maintenance preserveed this benefit. A shift from insulin sensitivity to insulin resistance increased oxLDL. Smokers had more oxLDL than non-smokers. A combination of obesity and smoking, or smoking and low serum total testosterone,resulted in even higher levels of oxLDL than any of the three conditions alone. Proportioning oxLDL to HDL-c or apoA1 stood out as a risk factor of all-cause mortality in the elderly. Conclusions: OxLDL was associated with aging, androgens, smoking, obesity, insulin metabolism, weight balance and other circulating lipid classes. Through this variety of metabolic environments containing both constant conditions (aging and gender) as well as lifestyle issues, these findings supported an essential and multidimensional role that oxLDL plays in atherosclerosis pathogenesis.

Biomarkers, medical treatment and fetal intrapartum surveillance in intrahepatic cholestasis of pregnancy

Intrahepatic cholestasis of pregnancy (ICP) is a pregnancy-specific disorder characterized by maternal pruritus and elevated liver enzymes. It usually begins in the third trimester of pregnancy and resolves spontaneously after delivery. ICP is considered benign for the pregnant woman, but it is associated with an increased risk for unexplained term stillbirth and preterm delivery. There are no specific laboratory markers to diagnose ICP. The diagnosis is currently based on the presence of maternal pruritus and elevated values of alanine aminotransaminases (ALT) and serum bile acids (BA). Recently, ursodeoxycholic acid (UDCA) has been used for treatment. Mechanisms leading to intrauterine fetal death (IUFD) may be multifactorial and are unknown at present. For this thesis, 415 pregnant women with ICP were studied. The aim was to evaluate the value of the liver enzyme glutathione S-transferase alpha (GSTA) as a specific marker of ICP and to assess the effect of maternal UDCA therapy on maternal laboratory values and fetal outcome. The specific markers predisposing the fetus to heart arrhythmia were studied by comparing waveform analysis of fetal electrocardiograms (FECG) during labor in pregnancies complicated by ICP with controls. The levels of maternal GSTA were high and the values correlated with the value of ALT in patients with ICP. UDCA therapy reduced the values of the liver enzymes and alleviated maternal pruritus, but it did not influence maternal hormonal values. Although the newborns experienced an uneventful perinatal outcome, severe ICP was still associated with preterm birth and admission to the neonatal intensive care unit (NICU). There were no significant differences in intrapartum FECG findings between fetuses born to ICP women and controls.

Diel patterns and tissue-specificity of environmental responses in fish

Humans are profoundly changing aquatic environments through climate change and the release of nutrients and chemicals. To understand the effects of these changes on natural populations, knowledge on individuals’ environmental responses is needed. At the molecular level, the environmental responses are partly mediated by chances in messenger RNA and protein levels. In this thesis I study messenger RNA and protein responses to an assortment of environmental stressors in fish. As daily (diel) rhythms are known to be ubiquitous in different tissues, I particularly focus on diel patterns in the responses. The studied species are the three-spined stickleback (Gasterosteus aculeatus L.) and the Arctic char (Salvelinus alpinus L.), both of which have circumpolar distribution in the Northern hemisphere. In the first two studies, three-spined sticklebacks were exposed to both the non-steroidal anti-inflammatory drug diclofenac and low-oxygen conditions (hypoxia), and their responses measured at separate time points in the liver and gills. The results show how the seemingly unrelated environmental stressors, hypoxia and anti-inflammatory drugs, can have harmful combined effects that differ from the effects of each stressor alone. Moreover, both stressors disturbed natural diel patterns in gene expression. In the third study, I studied the responses of three-spined sticklebacks to two test chemicals: one used in hormonal medicine (17α-ethinyl-oestradiol) and one used as a plasticizer and solvent chemical (di-n-butyl phthalate). The results suggest that the phthalate can affect genes related to spermatogenesis in fish testes, while estrogen-mimicking compounds can lead to numerous disturbances in the endocrine system. In the final study, the temperature-dependence of diel rhythms in messenger RNA levels were evaluated in the liver tissue of the Arctic char, a cold-adapted salmonid. The results show that cold acclimation repressed diel rhythms in gene expression compared to warm-acclimated fish, in which the expression of hundreds of genes was rhythmic, suggesting the circadian clock of the Arctic fish species can be sensitive to temperature. Overall, the results of the thesis indicate that fishes’ responses to abiotic factors interact with their diel rhythms, and more studies on the consequences of these interactions are needed to comprehensively understand human impacts on ecosystems.