I. Patterns of gametogenesis and gonadogenesis of mammals and humans
- Fundamental issues of chronology, dynamics and patterns of development of male and female gonads and germ cells of humans and several species of mammals (cows, green monkeys, pigs, rats, mice) have been studied.
- Original schemes of oogenesis and spermatogenesis have been created (Kurilo L.F., 1983, 1989–85, 1989, 1996, which are included in relevant textbooks and monographs)
- They were first to prove with the use of quantitative method, that the formation of primordial follicles (for example, in humans, cows, mice, and rats) occurred after the completion of conjugation (in zygotene) and crossing-over (in pachytene): around oocytes at the stage of diplonema meiosis prophase 1 (Kurilo, 1980 1985; Kurilo, 1981; Teplyakova, Kurilo et al., 1984).
- First in the world a quantitative method traced the development of stages: the preleptotene condensation and de-condensation of chromosomes in oocytes in humans, cows, mice and rats, and in male germ cells of the embryo and the human fetus; condensation of chromosomes and their localization around the nucleolus (karyosphere stage) in oocytes before diakinesis from maturing follicles in humans, cows, green monkeys (Kurilo, 1981; 1982, 1984, 1985; Kurilo et al. 1983; 1986).
- Bragina E.E., Doctor of biological science, developed a method of sperm electron microscopic analysis (SEMA). The introduction of SEMA into the practice of andrological and medical-genetic examination of patients with reproduction disorders made it possible to obtain significantly new data on the nature of male fertility disorder and idiopathic infertility. SEMA made it possible to detect abnormalities in spermatozoa that were not previously available with light-optical microscopy: it was possible to establish the nature of microstructural anomalies of gametes, in particular, such forms of teratozoospermia as anomalies of the centrioles and chromatin structure, to detect intracellular viral infection.
II. Development of a testing system for gametotoxic effect, schemes and quantitative criteria for its evaluation (for oogenesis and spermatogenesis)
Kurilo L.F. developed a new direction in gameology, namely, the quantitative testing of gametes state according to the stages of their development (gametogenesis). The author's methods, systems and criteria for quantifying gamete-and gonadotoxic effects (physical, chemical and biological factors) on male and female sex cells and sex glands of humans and animals have been developed for this area of research. These methods are used in diagnosing the causes of infertility and gametopathies, in testing the damaging gonad and gamete factor, in analyzing the effectiveness of gametogenesis disorders treatment, as well as in preventing these conditions. They showed the damage of oogenesis and spermatogenesis in the offspring of the first generation (F1) of mice and rats after administration of a number of toxic compounds to pregnant females (ThioTEPA, alcohol, nicotine, oxytetracycline), and discovered the prolonged effect of the examined compounds on the oogenesis and spermatogenesis F1. Patents were obtained for three of the developed methods: 1) Kurilo L.F. "Method of cytological diagnosis of spermatogenesis disorders", patent for invention No. 2328736 of 1/2-2007; 2) Kurilo L.F. "Method for ovarian generative potency diagnosis ", patent for invention No. 2367949 of 03/4-2008; 3) Korolev Yu.N., Kurilo L.F., Nikulina L.A., Panova L.N., Geniatulina M.S., Shileiko L.V. "Method of secondary prevention of spermatogenesis disorders when exposed to radiation" patent for invention No. 2169571.
III. Analysis of the structure, causes and developmental mechanisms of the human reproductive system hereditary pathology
- Determination of frequency and types of chromosomal aberrations (CA) of sex chromosomes and autosomes (in peripheral blood lymphocytes and germ cells) for infertility in men and women, for disorders of sexual development in children of both genders.
- Identification of frequency and types of the long arm of Y-chromosome microdeletions (at Yq11.2 locus (AZF, Azoospermia factor) - azoospermia factor) in Russian men with infertility associated with various forms of impaired spermatogenesis and pathozoospermia (in particular, with developmental non-obstructive forms of azoospermia and severe oligozoospermia).
- Analysis of mutations and polymorphisms of some genes that control the development and function of the reproductive system (AR/HUMARA, CFTR) in patients with various forms of reproductive system disorders, and in fertile men.
- Analysis of genetic causes for impaired differentiation of the reproductive system in humans (at different levels of gender formation). Research on the relationship between genetic changes (chromosomal abnormalities, microstructural chromosome rearrangements, gene mutations and polymorphisms) associated with reproductive disorders and the phenotype in syndromic and non-syndromic abnormality of the reproductive system (cystic fibrosis, CBAVD syndrome, Klinefelter syndrome, Shereshevsky-Turner syndrome, de la Chapelle syndrome/46-XX-male, ovotestichulyarnaya form of sex development disorder, various forms of gonadal dysgenesis, gonosomic mosaicism and other genetically determined reproductive system disorders).
- Examination of nature and patterns of sex chromosomes (gonosome) various mutations and gonosomic mosaicism in various cell lines, genotyphenotypic relationship of sex development anomalies and reproductive system development, gametogenesis and fertility disorders in patients with some numerical, structural and microstructural aberrations of their sex chromosomes, their impacts on reproduction and human fertility.
- Development of new methods of molecular genetic diagnostics (chromosomal abnormalities and gene mutations) of disorders of reproductive system development and function in humans (jointly with DNA-diagnostics Lab., FSBI RCMG, Laboratory head is Prof. Polyakov A.V., Doctor of biological science).
- Study of meiosis process and formation of a synaptonemal complex in patients with impaired spermatogenesis and male infertility, including those associated with genetic causes (chromosomal abnormalities, microdeletions of the Y chromosome). Elaboration of approaches to identification of human meiotic mutations (jointly with the Cytogenetics Laboratory of the Research Institute of General Genetics named after N.I. Vavilov of RAS, Laboratory head is Prof. Kolomiets O.L., Dr. of biological science)
- Analysis of the impact of chromosomal aberrations (CA) in ejaculate germ cells on gametogenesis and fertility. Estimation of chromosomes nondisjunction frequency in gametes of karyotype constitutive anomalies carriers (balanced translocations, inversions and supernumerary marker chromosomes) with different fertility status (jointly with the Cytogenetics Laboratory of the FSBI RCMG, Laboratory head is Shilova N.V. Dr. of medical science).
- Identification of specific anomalies of various gamete ultrastructures (chromatin and DNA of sperm cells, acrosomes, flagellum, fibrous layer, mitochondria, etc.) and somatic cells of male and female reproductive systems in certain forms of male and female infertility. Among them are the absence of dynein arms or central duplet of microtubules axonemes of spermatozoa in primary ciliary dyskinesia and Kartagener syndrome; chromatin condensation disorder in "immature" chromatin syndrome, globuzoospermia (sperm cell acrosome) and sperm fibrosis dysplasia syndrome, Sertoli cells abnormalities in severe forms of pathozoospermia, and endometrial transformation abnormalities in embryonic implantation disorders.
- Analysis of genital infections (viral, bacterial, protozoal) impact on gametogenesis and reproductive function (including male and female infertility and miscarriage). We were the first in the world to identify intra-gamete viral infection in male germ cells.
- Establishing of a database (electronic resources) on genetically determined diseases of the reproductive system.
IV. Study of moral, ethical, and legal issues in genetics and human reproduction, and the use of biomedical technologies.
- The degree of awareness of the population, specialists in medical fertility science and patients (with childbearing problems) in the ethical and legal issues of human reproduction (by means of a survey).
- Ethical and legal issues in the use of reproductive and other biomedical technologies.
- “Human embryo status” – international and domestic (Russian) legislation.
- In 1997-2003, Kurilo L.F. has been an expert of the working group on protection of human embryos and fetuses (CDBI-co-GT3) under the Council of Europe's Steering Committee on Bioethics. A Report has been published (the title is given in the References).
- Kurilo L.F. is one of co-authors of the Federal Law on Temporary Prohibition of Human Cloning (2002). Since 2011, this law has been extended indefinitely.