Vladimir Isachenko was awarded a PhD in the field of human and animal physiology from Kharkov Institute in the former USSR. He has been working in the area of reproductive biology since 1985, beginning with studies on the oocytes and embryos of agricultural and laboratory animals before moving on to study humans in 1990. Now his main work focuses on the development of new methods of preservation of the genome of cancer patients through vitrification of spermatozoa, oocytes, follicles and ovarian tissue. His specific area of interest is the effect of cold on the genome. Abstract This investigation compared conventional freezing of human ovarian tissue using either spontaneous or initiated ('seeded') ice formation. Biopsies of ovarian tissue were obtained from women with indications for chemotherapy or radiotherapy. Small pieces of experimental tissue were randomly distributed into three groups that were then subjected to different treatments prior to culture in vitro for 16 days: the control group, no treatment, cultured immediately after biopsy (group 1); cryopreservation/ thawing with spontaneous ice formation (group 2); and cryopreservation/thawing with initiated ice formation (group 3). Follicle viability and hormonal activity were then evaluated. There was no significant difference between groups regarding the concentration of oestradiol 17-β in the culture supernatant, whereas progesterone concentration was significantly (P < 0.05) higher in group 1 compared with group 2 or 3. There was a significant (P < 0.05) difference in primordial and primary follicle density between all of the groups (group 1 having the highest and group 2 having the lowest) and group 2 had significantly (P < 0.05) fewer normal grade follicles than the other two groups. For optimal cryopreservation of human ovarian tissue, the protocol of conventional freezing should therefore include a step of initiated ice formation.
Human milk (HM) contains antioxidant enzymes such as superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase. Melatonin (MT) shows antioxidative properties both directly by stimulating the activities of antioxidant enzymes and indirectly by scavenging free radicals. The aim of this study was to evaluate the effect of MT status on the level of SOD, GSH-Px 3 and the total antioxidant capacity (TAC) of HM.
Material and Methods
114 samples of HM were collected from 22 healthy women during daytime (DT; 10a.m.-9.59p.m.) and nighttime (NT; 10p.m.-9.59a.m.). MT, SOD, GSH-Px3 levels and TAC were assayed and compared between DT-HM and NT-HM. Moreover, the Pearson’s correlation coefficient was calculated between MT and SOD, GSH-Px3 and TAC.
Results
MT showed a circadian rhythm with high levels at NT and low levels at DT (mean± SEM; 9.2±1.2 vs. 2.0±0.3 pg/ml, respectively, p<0.0001), while SOD, GSH-Px3 and TAC had no circadian changes in HM (p>0.05). There was no correlation between MT levels and SOD, GSH-Px3 levels and TAC.
Conclusion
The MT status influence neither SOD and GSH-Px3 levels nor TAC of HM. It should be pointed that our results represent only the effect of MT on the extracellular component of the antioxidant system and it does not exclude the possibility that high MT concentrations in HM may induce the intracellular and mitochondrial antioxidant enzyme system in the gastrointestinal tract.
The aim of this study was to evaluate the influence of different maternal and fetal albumin concentrations on the transplacental transfer and the placental tissue accumulation of digoxin. Digoxin passage across the isolated lobules of 15 human placentae was calculated from repeated fetal and maternal perfusate samples, and placental tissue digoxin concentrations were measured at the end of the experiments. Metildigoxin (Lanitop) was added to the maternal medium at a concentration of 5.70 +/- 0.73 ng mL-1, and maternal and fetal perfusate albumin (BSA) concentrations were kept equal either at a high concentration of 21 g L-1 (Group I; n = 5) or at a low concentration of 3 g L-1 (Group III; n = 5), or differed with a materno-fetal gradient of 21:3 g L-1 (Group II; n = 5). In the experiments with low maternal albumin concentrations (Group III), digoxin concentrations in the maternal circuit decreased to 3.56 ng mL-1, whereas digoxin concentrations in the fetal circuit reached 2.59 ng mL-1 over a 3-h period. With maternal BSA concentrations of 21 g L-1 (Group I and Group II), the decrease in digoxin concentration in the maternal circuit was lower (P < 0.05), and digoxin tissue concentrations at the end of the experiments were smaller (0.45 +/- 0.07 and 0.42 +/- 0.03 v. 0.82 +/- 0.32 ng mg-1 protein, Group I and Group II v. Group III respectively; P < 0.05). Comparing only those lobules with similar high concentrations of maternal protein, fetal BSA concentrations of 21 g L-1 resulted in a greater increase in digoxin concentrations in the fetal circuit (end-feto to initial-maternal digoxin concentrations of 0.44 +/- 0.08 v. 0.37 +/- 0.04 ng mg-1 protein (Group I v. Group II respectively), although this was not significant. The data suggest that maternal and fetal serum albumin concentrations may have an influence on transplacental digoxin transfer, and this should be considered when treating fetuses with cardiac disease transplacentally with glycosides.
How do anti-Müllerian hormone (AMH) serum concentrations and follicle densities (FDs) change with age and disease and what are the implications for fertility preservation?AMH concentrations and FD do not correlate in young women, and AMH but not FD is reduced in some diseases, limiting the value of AMH as a predictive parameter of ovarian tissue transplantation.AMH is widely used as a parameter to estimate the ovarian reserve. However, the reliability of AMH to predict total number of follicles and the FD is questionable. Women with lymphoma and leukaemia have been shown to have reduced AMH concentrations, but it is unknown if the FD is also reduced. In fertility preservation it is essential to estimate the correct total number of follicles and the FD, as ovarian tissue should only be cryopreserved if ovarian reserve is high. Furthermore, the amount of tissue to be transplanted should be based on the estimation of the real FD.This retrospective observational study included 830 women (mean ± SD age, 28.2 ± 6.81 years; range, 4-43 years) with malignant (n = 806) and benign (n = 24) diseases who cryopreserved tissue in a single centre as part of a national fertility preservation programme. Females with ovarian surgery or known predispositions for a reduced ovarian reserve were excluded. AMH concentrations and FD were evaluated from March 2011 to September 2016.AMH concentrations were analysed before gonadotoxic therapies. Standardized biopsies, obtained from different areas of ovarian cortex, were collected. FD was analysed after tissue digestion and calcein staining and was expressed as average number of primordial and primary follicles count per 3 mm biopsy and per cubic millimeter tissue. AMH concentrations and FD were analysed in relation to age and diagnosis group. Both parameters were age adjusted, and associations between the different diagnosis groups and AMH versus FD were assessed.Mean ± SD AMH concentration was 3.1 ± 2.81 g/ml, mean FD per 3 mm biopsy was 137 ± 173.9 and 19.4 ± 24.60 per mm3. Maximum AMH concentrations were found in children and teenagers at the age of 6-10 years (5.71 ng/ml) and in adults at the age of 21-25 years (3.33 ng/ml). FD was highest in young children up to an age of 15 years and decreased with increasing age. AMH and FD were not correlated in women ≤20 years and weakly to moderately correlated in women 21-40 years (r = 0.24-0.39). Age-adjusted correlations between AMH and FD were demonstrated in several diagnosis groups such as breast cancer, leukaemia, sarcoma, gastrointestinal cancer and gynaecological cancer but not in the groups exhibiting Hodgkin's and non-Hodgkin's lymphoma, cerebral cancer, other types of malignancies and other types of benign diseases. Further statistical analysis supported the finding that, in some diagnosis groups such as Hodgkin's lymphoma and in gynaecological cancer, AMH concentrations but not FDs are reduced, questioning the prognostic accuracy of AMH for the FD in these diseases.Even though biopsies were taken from different sites, heterogenous distribution of follicles might have had some effect on the accuracy of the analysis.AMH should be used with care to estimate the total ovarian reserve and FD of cancer patients in young women in some diseases. Therefore, calculating the amount of ovarian tissue to be transplanted based solely on AMH might be inaccurate whereas FD might be a better parameter.The study did not receive any exterior funding.
Abstract Very high concentrations of CA 125 have been found in some ovarian cancer patients after repeated radioimmunodetection with anti-CA 125 antibodies [OC125-F(ab')2]. In one patient we measured a CA 125 concentration of 135,000 kilo-arb. units/L, using an enzyme immunoassay involving OC125 antibodies. With an immunoradiometric assay involving use of two new anti-CA 125 antibodies (B43.13 and B27.1), the CA 125 concentration was 34 kilo-arb. units/L, indicating a discrepancy. The component responsible for the high result in the enzyme immunoassay could be purified by immunoaffinity chromatography on Protein A-Sepharose. Furthermore this component bound to anti-human IgG-Sepharose in the same manner as did the serum IgG fraction. Adsorption of human anti-mouse antibodies present in the serum did not decrease the CA-125-like material. Binding of whole OC125 antibodies to the purified CA-125-like material was inhibited completely in the presence of CA 125 antigen. We infer that the false-positive CA 125 activity is ascribable to a human IgG directed against an idiotope of the OC125 antibody, which was induced by repeated application of OC125 antibodies. To avoid falsely positive results in patients receiving OC125 antibodies, CA 125 should be measured by an assay in which other antibodies are used.
