Sriubienė, Margarita

Parazitologijos mokslas Lietuvoje turi ilgą ir reikšmingą istoriją, glaudžiai susijusią su šalies gamtos ypatumais, žemės ūkio tradicijomis ir visuomenės sveikatos poreikiais. Šiandienos pasaulyje, susiduriant su klimato kaita, globalizacija ir naujų infekcinių ligų atsiradimu, parazitologijos reikšmė Lietuvoje dar labiau padidėja. Klimato ir geografinės sąlygos sudaro palankias sąlygas veistis Įvairiems parazitams, plėstis parazitų pernešėjų arealams, o tai kelia naujus iššūkius visuomenės sveikatai ir biologinei Įvairovei. Parazitologijos studijų dalykas mūsų Universitete nuėjo ilgą evoliucijos kelią, keitėsi studijų programos, mokymo priemonės, gilėjo patirtis. Ši patirtis (buvusių ir esamų) Biologinių sistemų ir genetinių tyrimų instituto darbuotojų perduodama Jums, mieli skaitytojai. Mūsų parengta mokomoji knyga skiriama pirmojo kurso studentams, studijuojantiems biomedicinos mokslus - mediciną, odontologiją, burnos higieną, slaugą ir visuomenės sveikatą. Leidinyje pateikiamas parazitologijos kursas, apimantis svarbiausius žmogaus parazitus, jų morfologiją, sukeliamas ligas, diagnostikos principus ir profilaktikos priemones. Šias teorines žinias padės įtvirtinti praktikos darbai. Kiekvieno praktikos darbo pabaigoje pateikiami saviruošos klausimai, skirti suprasti ir Įsiminti praktikos darbų metu analizuojamus parazitus. Tikimės, kad šis leidinys bus naudingas ne tik pratybų metu, bet ir savarankiškai mokantis, ruošiantis atsiskaitymams arba rengiant baigiamuosius darbus.

Tai, kas atrodė neįmanoma prieš trisdešimt metų, tampa realybe. Šios mokomosios knygos autoriai mano, kad tikrai žengiame individualizuotos medicinos link. 1990 metais pradėtas žmogaus genomo sekoskaitos projektas. Nuo 2014 metų atliekama viso genomo sekoskaita atskiriems pacientams. Klinikine rutina tampa viso egzomo ar net genomo tyrimai. CRISPR/Cas9 genų redagavimas leidžia plačiau taikyti genų terapiją, o Lietuvoje genų terapija pradedami gydyti pirmieji pacientai. Nepaisant įspūdingų galimybių, norėtųsi, kad nepamirštume istorijos, todėl įžangą tęsiame tais pačiais žodžiais, kuriais prasidėjo 2006, 2009, 2013 ir 2021 metais parašytų „Genetikos pratybų“ pratarmės: „Genetikos dėstymo pradžia ne tik Lietuvos sveikatos mokslų universitete, bet ir Lietuvoje sietina su prof. Tado Ivanausko (1882-1970) vardu. Profesorius T. Ivanauskas į paskutinę semestro paskaitą atsinešdavo plakatus su „Mendelio žirneliais“ ir paaškindavo studentams „uždraustuosius paveldimumo dėsnius.“ Genetikos studijų dalykas mūsų Universitete nuėjo ilgą evoliucijos kelią. Vadovaujant buvusiems KMI (vėliau KMA ir KMU) Biologijos katedros vadovams doc. J. Lapinskaitei ir prof. A. Bertuliui, citogenetikos mokslo pradininkams Lietuvoje prof. A. Sinkui ir doc. L Andriuškevičiūtei, keitėsi studijų programos, mokymo priemonės, gilėjo patirtis. Ši patirtis (buvusių ir esamų) instituto darbuotojų ir yra perduodama Jums, mieli skaitytojai. Ši mokomoji knyga skiriama visiems, studijuojantiems mediciną. Bet pravers ir Slaugos, Odontologijos, Visuomenės sveikatos fakultetų studentams. Visi praktikos darbai peržiūrėti ir pataisyti pagal naują Medicinos fakulteto studentams patvirtintą programą. Kiekvieno praktikos darbo pabaigoje nurodyta pagrindinė ir papildoma literatūra, interneto tinklalapiai, užduotys, skirtos studentų praktikos darbams.

Purpose: This study aimed to evaluate the associations of GJD2 (rs634990, rs524952) and RASGRF1 (rs8027411, rs4778879, rs28412916) gene polymorphisms with refractive errors.Methods: The study included 373 subjects with refractive errors (48 myopia, 239 myopia with astigmatism, 14 hyperopia, and 72 hyperopia with astigmatism patients) and 104 ophthalmologically healthy subjects in the control group. A quantitative real-time polymerase chain reaction (qPCR) method was chosen for genotyping. Statistical calculations and analysis of results were performed with IBM SPSS Statistics 27 software.Results: The correlations in monozygotic (MZ) twin pairs were higher compared to DZ pairs, indicating genetic effects on hyperopia and astigmatism. The heritability (h2) of hyperopia and astigmatism was 0.654 for the right eye and 0.492 for the left eye. The GJD2 rs634990 TT genotype increased the incidence of hyperopia with astigmatism by 2.4-fold and the CT genotype decreased the incidence of hyperopia with astigmatism by 0.51-fold (p < 0.05). The GJD2 rs524952 AT genotype reduced the incidence of hyperopia with astigmatism by 0.53-fold (p < 0.05). Haplotype analysis of SNPs in the GJD2 gene revealed two statistically significant haplotypes: ACTAGG for rs634990 and TTTAGA for rs524952, which statistically significantly reduced the incidence of hyperopia and hyperopia with astigmatism by 0.41-fold (95% CI: 0.220-0.765) and 0.383-fold (95% CI: 0.199-0.737), respectively (p < 0.05). It was also found that, in the presence of haplotypes ACTAGG for rs634990 and TATAGA for rs524952, the possibility of hyperopia was reduced by 0.4-fold (p < 0.05).Conclusions: the heritability of hyperopia and hyperopia with astigmatism was 0.654-0.492, according to different eyes in patients between 20 and 40 years. The GJD2 rs634990 was identified as an SNP, which has significant associations with the co-occurrence of hyperopia and astigmatism. Patients with the GJD2 gene rs634990 TT genotype were found to have a 2.4-fold higher risk of develop hyperopia with astigmatism.

Introduction Most of our information about the world around us comes from our visual system. When this system functions well, relevant objects in our field of vision are imaged and focused on the retina. Refractive errors are among the most frequent treatable “diseases” [1]. Refractive errors occur when the refractive system of the eye fails to correctly focus rays of light from an object onto the retinal plane [2]. Refractive errors are the most common vision defect affecting all age groups and their number is always growing [3]. Knowledge of the prevalence of refractive errors and risk factors can help plan effective measures to reduce them. Aim To identify GJD2 (rs634990, rs524952) and RASGRF1 (rs8027411, rs4778879, rs28412916) gene variants in subjects with refractive errors and control subjects. To evaluate the significance of GJD2 (rs634990, rs524952) and RASGRF1 (rs8027411, rs4778879, rs28412916) gene variants for ocular refractive errors. To evaluate the associations of GJD2 (rs634990, rs524952) and RASGRF1 (rs8027411, rs4778879, rs28412916) gene variants with the degree of myopia and hyperopia. Methods The study included 373 individuals with refractive errors and 104 ophthalmologically healthy subjects. DNA was extracted from peripheral blood leukocytes using the salting- out method. The quantitative real-time polymerase chain reaction (qRT-PCR) method was chosen for genotyping. Statistical calculations and analysis of the results were performedusing ”IBM SPSS Statistics“ software.

Tai, kas atrodė fantastika prieš trisdešimt metų, tampa realybe – šios knygelės autoriai mano, kad tikrai žengiame individualizuotosios medicino link. 1990 metais pradėtas žmogaus genomo projektas truko trylika metų ir kainovo apie vieną milijardą dolerių. 2014 metais Europos Žmogaus genetikos kongrese Milane dr. S. Kingsmore'as pristatė pacientus, kuriems buvo įtariama reta genetinė liga. Jų genomo sekoskaita ir analizė truko dvi-šešias dienas ir kainavo apie 1000 dolerių. Tačiau turime nepamiršti istorijos ir įžangą tęsiame tais pačiais žodžiais, kuriais prasidėjo 2006, 2009 ir 2013 metais parašytos knygos "Genetikos pratybos pratarmės: "Genetikos dėstymo pradžia ne tik Lietuvos sveikatos mokslų universitete, bet ir Lietuvoje, sietina su prof. Tado Ivanausko (1882-1970) vardu. Profesorius T. Ivanauskas į paskutinę semestro paskaitą atsinešdavo plakatus su "Mendelio žirneliais" ir paiškindavo studentams uždraustuosius paveldimumo dėsnius". Genetikos studijų kursas mūsų Universitete nuėjo ilgą evoliucijos kelią. Vadovaujant buvusiems KMI (vėliau KMA ir KMU) Biologijos katedros vadovams doc. J. Lapinskaitei ir prof. A. Bertuliui, citogenetikos mokslo pradininkui Lietuvoje prof. A. Sinkui, keitėsi studijų programos, mokymo priemonės, gilėjo patirtis. Ši patirtis (buvusių ir esamų) instituto darbuotojų ir yra perduodama Jums, skaitytojai. Ši mokomoji knyga skiriama biomedicinos mokslus, pirmiausia mediciną, slaugą, odontologiją, visuomenės sveikatą studijuojantiems studentams. Visi praktikos darbai peržiūrėti ir pataisyti, nurodyta pagrindinė bei papildoma literatūra ir interneto tinklalapiai. Atsirado ir naujas praktikos darbas, kuriame sužaindinama su molekulinės genetikos metodais, taikomais genetinių ligų diagnostikai.

This laboratory activity book was designed at the Biology Systems and Genetic Research Institute of the Lithuanian University of Health Sciences for the first year biomedical science students of the Faculties of Medicine and Odontology. Together with the step-by-step guide for the parasitology labs, the workbook provides some theoretical background, which can also be used for individual studies, practical training, studying of other disciplines such as Pharmacy, Oral Hygiene, Occupational Therapy, Public Health, Nursing and Infectious Diseases and for the preparation of Bachelor's and Master's theses. We would like to thank Prof. Dalia Pangonytė and Dr. Milda Kuprytė (Clinic of Pathological Anatomy), Prof. Algimantas Kriščiukaitis and Dr. Robertas Petrolis (Department of Physics, Mathematics and Biophysics) and everyone who contributed to this publication with their insights, suggestions and technical support. Also we thank Rūta Slavinskienė for the translation of this whole book from Lithuanian into English language, and Mindaugas Mickis for the translation of one capter. The authors thank the students, teachers, and colleagues who helped to improve this workbook. We believe it would largely contribute to the development of other educational materials in the future.

PURPOSE: This study aimed to find associations between miR-328 expression in whole blood, polymorphism at 3'UTR of the PAX6 gene (paired box homeotic gene 6) and myopia. METHODS: We evaluated 451 individuals (142 individuals with low, 49 with moderate and 13 with high-degree myopia, and 247 healthy individuals). DNA and RNA were extracted from peripheral blood samples. Expression of miR-328 was assessed and genotyping of single-nucleotide polymorphisms (SNPs) of the PAX6 (rs662702) performed using the Applied Biosystems 7900HT Real-Time Polymerase Chain Reaction System. RESULTS: Moderate and high degree myopia showed significant differences between TT and CT genotypes of the PAX6 gene (p < 0.001). In the myopia group, 71.4% of the subjects had the TT genotype and 28.6% had the CT genotype; meanwhile in the control group, 97.1% had the TT genotype and 2.9% had the CT genotype. The odds ratio of having moderate and/or high degree myopia for individuals with the CT genotype was 13.6 (2.865-64.55) 95% CI versus TT genotype (p = 0.001). MiR-328 results showed that ∆Ct values differed statistically significantly between the myopia and control groups. Patients with myopia in the peripheral blood cells had a higher expression of miR-328 than controls (p < 0.05). CONCLUSIONS: Significant differences were detected between the PAX6 gene (rs662702) TT and CT genotypes in moderate and high degree myopia; the risk C allele increased the risk for myopia. The expression level of miR-328 in peripheral blood cells was higher in patients with myopia than controls. We did not find the association between expression of mir-328 in the peripheral blood cells and PAX6 gene (rs662702) polymorphism comparing myopia and control groups.

BACKGROUND: This study aimed to assess heritability of myopia in Lithuania and evaluate both genes GJD2 (Gap Junction Protein, Delta 2) and RASGRF1 (RAS protein-specific guanine nucleotide-releasing factor 1) relation with myopia. METHODS: In this study Lithuanian twin population aged between 18 and 40 (n = 460) were examined. Single-nucleotide polymorphisms of the RASGRF1 (rs8027411) and GJD2 (rs634990) genes were assessed by real-time polymerase chain reaction method. RESULTS: Intrapair correlations for spherical equivalent in all twin pairs were significantly higher in MZ twin pairs r = 0.539 (p < 0.001, 95% CI 0.353-0.684) than in DZ twin pairs r = 0.203 (p < 0.01, 95% CI 0.0633-0.442) in myopia group. Correlations for spherical equivalent in emmetropia group were not significant in MZ twin pairs r = 0.091 (p > 0.05, 95% CI -0.215-0.381) and in DZ twin pairs r = - 0.220 (p > 0.05, 95% CI -0.587-0.222). The odds ratio (95% CI) were 2.7 (1.018-7.460) for combinations of genotypes of rs634990 CC and rs8027411 GT (p = 0.046). CONCLUSIONS: Our studies have shown that the heritability of myopia makes 67.2% in Lithuania. Persons with combinations of genotypes rs634990 CC and rs8027411 GT have 2.7 times higher odds to have myopia.

Background. Myopia is a significant and increasing public health problem over time. It′s known that genetic factors in disease development are important [1]. A genomewide linkage study reveals that PAX6 gene is a candidate gene for the development of myopia [2, 3]. It′s reported that the 3′UTR single nucleotide polymorphism (SNP) rs662702 of the PAX6 is associated with extreme myopia [4]. This study aimed to find associations between polymorphism at 3‵UTR of the PAX6 gene (rs662702) and myopia in Lithuania. Methods. We evaluated 418 individuals (195 with myopia and 223 healthy). The study participants were between the ages of 18 and 40 years. SNP of the PAX6 were assessed by using the Applied Biosystems 7900HT Real-Time Polymerase Chain Reaction System. Results. We found that 90.3% of the subjects had TT genotype, CT – 9.69%, and only one had a CC genotype in the miopia group with low, moderate and high degree. In the control group, 87.9% of the subjects had TT genotype and 12.1 % CT. Repeated calculations by separating only moderate and high degree myopia showed significant differences between CC and CT genotypes of PAX6 gene (p<0.001). 71.4% of the subjects had TT genotype, 28.6% CT and 97.1% TT genotype and 2.9% CT in myopia and control groups, respectively. The odds ratio having moderate and high degree myopia for individuals with a CT genotype was 13.6 (2.865-64.55) 95% C.I. versus TT genotype (p=0.001). Conclusions. Significant differences between the PAX6 gene TT and CT genotypes were detected only higher degree myopia. The risk C allele increases the risk for myopia.

Ši Lietuvos sveikatos mokslų universiteto Biologinių sistemų ir genetinių tyrimų instituto mokomoji knyga skiriama studijuojantiems biomedicinos mokslus – mediciną, odontologiją, burnos higieną, visuomenės sveikatą, slaugą – studentams. Leidinyje aprašomas parazitologijos ir ląstelės biologijos pratybų turinys bei pateikiama ir tam tikra teorinių žinių dalis, kuri gali būti naudinga ir pratyboms, ir savarankiškoms studijoms, ir praktikai bei studijuojant kitas disciplinas, pvz., farmaciją, ergoterapiją, kineziterapiją, užkrečiamąsias ligas ir pan., taip pat rengiant bakalauro bei magistro darbus.