Razvoj radiomarkiranog β-humanog koriogonadotropina

-human chorionic gonadotropin (-hCG) was successively labeled with [67Ga] gallium chloride after conjugation with freshly prepared diethylenetriaminepentaacetic acid dianhydride (ccDTPA). After solid phase purification of the radiolabeled hormone, high performance liquid chromatography showed radiochemical purity higher than 95 % under optimized conditions (specific activity = 2223 TBq mM1, labeling efficiency 80 %). Preliminary in vivo studies (ID g1, %) in male wild-type rats showed marked gonadal uptake of the tracer after 240 minutes in agreement with the biodistribution studies and reported -hCG receptors. Target to blood ratios were 5.1 and 15.2 after 3 and 24 hours, respectively, while target to muscle ratios were 35 and 40 after 3 and 24 hours, respectively.Beta-humani korionski gonadotropin (beta-hCG) uspješno je markiran s [67Ga] galijevim kloridom nakon konjugacije sa svježe priređenim dianhidridom dietilentriaminpentaoctene kiseline (ccDTPA). Nakon čišćenja radiomarkiranog hormona na čvrstoj fazi, radiokemijska čistoća bila je prema HPLC veća od 95 % (specifična aktivnost = 22-23 TBq mM-1, učinkovitost markiranja 80 %). Preliminarni in vivo pokusi (ID g-1, %) na mužjacima divljeg tipa štakora pokazali su da obilježeni hormon značajno ulazi u gonade nakon 240 minuta, što je u suglasnosti s ispitivanjima biodistribucije i podacima o receptorima za beta-hCG. Omjer koncentracija u gonadama i krvi bio je 5,1, odnosno 15,2 nakon 3, odnosno 24 sata, dok je omjer koncentracija u gonadama i mišićima bio 35, odnosno 40 nakon 3, odnosno 24 sata

Early Developing Pig Embryos Mediate Their Own Environment in the Maternal Tract

The maternal tract plays a critical role in the success of early embryonic development providing an optimal environment for establishment and maintenance of pregnancy. Preparation of this environment requires an intimate dialogue between the embryo and her mother. However, many intriguing aspects remain unknown in this unique communication system. To advance our understanding of the process by which a blastocyst is accepted by the endometrium and better address the clinical challenges of infertility and pregnancy failure, it is imperative to decipher this complex molecular dialogue. The objective of the present work is to define the local response of the maternal tract towards the embryo during the earliest stages of pregnancy. We used a novel in vivo experimental model that eliminated genetic variability and individual differences, followed by Affymetrix microarray to identify the signals involved in this embryo-maternal dialogue. Using laparoscopic insemination one oviduct of a sow was inseminated with spermatozoa and the contralateral oviduct was injected with diluent. This model allowed us to obtain samples from the oviduct and the tip of the uterine horn containing either embryos or oocytes from the same sow. Microarray analysis showed that most of the transcripts differentially expressed were down-regulated in the uterine horn in response to blastocysts when compared to oocytes. Many of the transcripts altered in response to the embryo in the uterine horn were related to the immune system. We used an in silico mathematical model to demonstrate the role of the embryo as a modulator of the immune system. This model revealed that relatively modest changes induced by the presence of the embryo could modulate the maternal immune response. These findings suggested that the presence of the embryo might regulate the immune system in the maternal tract to allow the refractory uterus to tolerate the embryo and support its development

mRNA and protein expression of FGF-1, FGF-2 and their receptors in the porcine umbilical cord during pregnancy.

The fibroblast growth factors (FGFs) are multifunctional proteins that, among other roles, regulate structural reorganization of uterine and placental vascular bed during pregnancy. Thus, we analyzed mRNA and protein expression and immunohistochemical localization of FGF-1 and FGF-2, and their receptors (FGFR-1 and FGFR-2) in the developing umbilical cord (UC) on days 40, 60, 75 and 90 of pregnancy and after the physiological delivery in the pig (day 114). qPCR analysis demonstrated an increase in FGF-1 and FGF-2 mRNA levels beginning on day 75 and on day 114 of pregnancy, respectively. In addition, significantly increased FGFR-1IIIc mRNA expression was also found on day 114. On the other hand, no significant changes in FGFR-2IIIb mRNA expression were observed. Western Blot analysis revealed a decrease in FGF-1 and FGFR-2 protein expression after day 40. Beside an increased protein expression of FGF-2 on day 60, no significant changes in FGFR-1 protein expression were detected. Immunohistochemical staining enabled detection of FGF-FGFR system, with different intensity of immunoreaction in endothelial and tunica media cells of the umbilical vessels and in allantoic duct and amniotic epithelium as well as in myofibroblasts. In conclusion, our results show that members of FGF-FGFR system are expressed specifically in UC structures. Furthermore their day of pregnancy-related expression suggest that they may be an important players during UC formation and development

Effect of vaginal administration of prostaglandin E2 and/or 17Betta-estradiol on luteal function and histological characteristics of the cervix in cyclic pigs

The overall objective of this study was to examine the effect of vaginal administration of prostaglandin E₂ (PGE₂) and/or17β-estradiol (E₂) on luteal function maintenance and histological properties of the porcine cervix. For this purpose, crossbred gilts were divided into three groups (n=5 per group) supplied on days 11-16 of the estrous cycle with suppositories containing: (1) placebo (Group I, Control); (2) 0.4 mg of E₂ (Group II); (3) 0.4 mg of E₂ and 2 mg of PGE₂ (Group III). Blood samples were collected on days 11-19 of the estrous cycle to determine the concentration of progesterone (P₄). Additionally, to examine local effects of the hormones applied, segments from the uterine and vaginal parts of the cervix and from the ovaries were collected post-mortem. Prolonged luteal function and extended synthesis of P₄ were observed in 2 of 5 gilts receiving PGE₂ and E₂ simultaneously (Group III). Then, these gilts were subdivided into Group IIIA (n=2; presence of corpora lutea on the ovaries) and Group IIIB (n=3; lack of corpora lutea). Increased levels of plasma P₄ were observed in Group IIIA on days 15-19 compared to Group IIIB and on days 16-19 compared to Group I and Group II (P<0.05; P<0.01; P<0.001, respectively). In the cervix of gilts in Groups II and III, enlarged blood vessels in the lamina propria of both parts of the cervix were observed. Furthermore, in Group II the epithelium of the uterine part of the cervix was thicker (P<0.001). Our study confirmed the proposed luteotrophic/antiluteolytic actions of E₂ and PGE₂ applied intravaginally. These results are significant considering that very low doses of E₂ were used when compared to previous attempts. Despite the inadequate response to treatments in some of the gilts, the local effects of these hormones on the histological properties of the porcine cervix suggest that further improvements in the vaginal administration route might help to elaborate new methods for enhancing the luteal function in the pig

Effect of carazolol on the oestrous behaviour and concentrations of luteinising hormone and steroids in lactating cows during the periovulatory period

Twelve multiparous, cycling, lactating Holstein-Friesian cows were synchronised with prostaglandin F2α and treated with either 2-5 mg carazolol or saline. There were no differences between the peripheral blood concentrations of oestradiol or progesterone, but in the cows treated with carazolol the periovulatory surge of luteinising hormone was delayed, and oestrous behaviour was expressed later than in the control cows