Effect of a high-calorie diet and constant light exposure on female reproduction, metabolism and immune inflammation: A comparative study of different mouse models.

PROBLEM: Excess caloric intake and irregular circadian rhythm could severely impair female reproductive, metabolic, and immune function. However, the similarities and differences between their individual and combined effects and mechanisms have not been fully elucidated. Due to limitations and confounding factors in clinical research, we used these two kinds of unhealthy factors to intervene the mice singly or in combination to explore their effects on individuals. METHOD OF STUDY: We used a high-calorie diet (HCD), constant light exposure (CLE), and a high-calorie diet combined with constant light exposure (HCD + CLE) to build three different mouse models. During the 9 weeks modeling period, the estrous cycles were monitored, and after modeling, the indicators of glycolipid metabolism, inflammation, and reproductive endocrine function were tested. RESULTS: We found that both HCD and CLE alone could induce ovulatory disorders, obesity, and chronic low-grade inflammation and inhibit melatonin secretion. The difference was that HCD significantly reduced the serum luteinizing hormone (LH) and testosterone (T) levels, inhibited the expression of FSH ß and LH ß in pituitary, increased cytochrome P450 enzymes and LH receptor expression in ovary, as well causing impaired glucose tolerance and hyperlipidemia, and significantly promoted the secretion of leptin and inhibited the secretion of adiponectin. However, CLE significantly increased blood LH and T, prompted the expression of kisspeptin in hypothalamus and LH ß in pituitary, and had no effect on glycolipid metabolic indexes or the secretion of leptin or adiponectin. The phenotype of HCD + CLE model was basically the same as that of HCD model, associated with more severe visceral obesity and chronic inflammation. CONCLUSIONS: In conclusion, we found that unhealthy lifestyle determines the phenotype of reproductive endocrine, immune, and metabolic disorders. These findings can provide theoretical support for the subsequent study of PCOS-like features.

Evaluation of mitochondria in oocytes following γ-irradiation.

Standard cytotoxic cancer treatments, such as radiation, can damage and deplete the supply of oocytes stored within the ovary, which predisposes females to infertility and premature menopause later in life. The mechanisms by which radiation induces oocyte damage have not been completely elucidated. The objective of this study was to determine if γ-irradiation changes mitochondrial characteristics in oocytes, possibly contributing to a reduction in oocyte number and quality. Immature oocytes were collected from postnatal day (PN) 9-11 C57Bl6 mice 3, 6 and 24 hours after 0.1 Gy γ-irradiation to monitor acute mitochondrial changes. Oocytes were classified as small (>20 µm) or growing (40-60 µm). Mitochondrial membrane potential was lost in 20% and 44% of small oocytes (~20 µm) at 3 and 6 hours after γ-irradiation, respectively, consistent with the induction of apoptosis. However, mitochondrial mass, distribution and membrane potential in the surviving small oocytes were similar to the non-irradiated controls at both time points. At 24 hours after γ-irradiation, all mitochondrial parameters analysed within immature oocytes were similar to untreated controls. Mitochondrial parameters within growing oocytes were also similar to untreated controls. When mice were superovulated more than 3 weeks after γ-irradiation, there was a significant reduction in the number of mature oocytes harvested compared to controls (Control 18 ± 1 vs 0.1 Gy 4 ± 1, n = 6/16 mice, p < 0.05). There was a slight reduction in mitochondrial mass in mature oocytes after γ-irradiation, though mitochondrial localization, mtDNA copy number and ATP levels were similar between groups. In summary, this study shows that γ-irradiation of pre-pubertal mice is associated with loss of mitochondrial membrane potential in a significant proportion of small immature oocytes and a reduction in the number of mature oocytes harvested from adult mice. Furthermore, these results suggest that immature oocytes that survive γ-irradiation and develop through to ovulation contain mitochondria with normal characteristics. Whether the oocytes that survive radiation and eventually undergo meiosis can support fertility remains to be determined.

Light-exposure at night impairs mouse ovary development via cell apoptosis and DNA damage.

The alternation of light and dark rhythm causes a series of physiological, biochemical and metabolic changes in animals, which also alters the growth and development of animals, and feeding, migration, reproduction and other behavioral activities. In recent years, many studies have reported the effects of long-term (more than 6 weeks) illumination on ovarian growth and development. In the present study, we observed the damage, repair and apoptosis of ovarian DNA in a short period of illumination. The results showed that, in short time (less than 2 weeks) illumination conditions, the 24-h light treatment caused the reduction of total ovarian follicle number and down-regulation of circadian clock related genes. Furthermore, the changed levels of serum sex hormones were also detected after 24-h light exposure, of which the concentrations of LH (luteinizing hormone), FSH (follicle-stimulating hormone) and E2 (estradiol) were increased, but the concentration of PROG (progesterone) was decreased. Moreover, 24-h light exposure increased the expression of DNA damage and repair related genes, the number of TUNEL and RAD51 positive cells. These results indicated that 24-h light exposure for 4, 8 and 12 days increased DNA damage and cell apoptosis, thereby affecting the development of ovary.

Folliculogenic factors in photoregressed ovaries: Differences in mRNA expression in early compared to late follicle development.

The early stages of ovarian folliculogenesis generally progress independent of gonadotropins, whereas later stages require signaling initiated by FSH. In Siberian hamsters, cycles of folliculogenesis are mediated by changes in photoperiod which depress the hypothalamic pituitary gonadal axis. Reduced gonadotropins lead to decreases in mature follicle development and ovulation; however, early stages of folliculogenesis have not been explored in regressed ovaries. We hypothesized that intraovarian factors that contribute predominantly to later stages of folliculogenesis would react to changes in photoperiod, whereas factors contributing to earlier stages would not change. To probe if the early stages of folliculogenesis continue in the photoinhibited ovary while late stages decline, we measured the mRNA abundance of factors that interact with FSH signaling (Fshr, Igf1, Cox2) and factors that can function independently of FSH (c-Kit, Kitl, Foxo3, Figla, Nobox, Sohlh1, Lhx8). While plasma FSH, antral follicles, and corpora lutea numbers declined with exposure to inhibitory photoperiod, the numbers of primordial, primary, and secondary follicles did not change. Expression of factors that interact with FSH signaling changed with changes in photoperiod; however, expression of factors that do not interact with FSH were not significantly altered. These results suggest that the photoinhibited ovary is not completely quiescent, as factors important for follicle selection and early follicle growth are still expressed in regressed ovaries. Instead, the lack of gonadotropin support that characterizes the non-breeding season appears to inhibit only final stages of folliculogenesis in Siberian hamsters.

Estudios ultrasonográficos diagnósticos y de control de los ciclos en reproducción asistida / No disponible

No disponible

Very low doses of heavy oxygen ion radiation induce premature ovarian failure.

Astronauts are exposed to charged particles during space travel, and charged particles are also used for cancer radiotherapy. Premature ovarian failure is a well-known side effect of conventional, low linear energy transfer (LET) cancer radiotherapy, but little is known about the effects of high LET charged particles on the ovary. We hypothesized that lower LET (16.5 keV/µm) oxygen particles would be less damaging to the ovary than we previously found for iron (LET = 179 keV/µm). Adult female mice were irradiated with 0, 5, 30 or 50 cGy oxygen ions or 50 cGy oxygen plus dietary supplementation with the antioxidant alpha lipoic acid (ALA). Six-hour after irradiation, percentages of ovarian follicles immunopositive for γH2AX, a marker of DNA double strand breaks, 4-HNE, a marker of oxidative lipid damage and BBC3 (PUMA), a proapoptotic BCL-2 family protein, were dose dependently increased in irradiated mice compared to controls. One week after irradiation, numbers of primordial, primary and secondary follicles per ovary were dose dependently decreased, with complete absence of follicles in the 50 cGy groups. The ED50 for primordial follicle destruction was 4.6 cGy for oxygen compared to 27.5 cGy for iron in our previous study. Serum FSH and LH concentrations were significantly elevated in 50 cGy groups at 8 week. Supplementation with ALA mitigated the early effects, but not the ultimate depletion of ovarian follicles. In conclusion, oxygen charged particles are even more potent inducers of ovarian follicle depletion than charged iron particles, raising concern for premature ovarian failure in astronauts exposed to both particles during space travel.

Environmental impacts on the gonadotropic system in female Atlantic salmon (Salmo salar) during vitellogenesis: Photothermal effects on pituitary gonadotropins, ovarian gonadotropin receptor expression, plasma sex steroids and oocyte growth.

The gonadotropic system and ovarian growth and development were studied during vitellogenesis in female Atlantic salmon subjected to either simulated natural photoperiod and ambient water temperature (NL-amb), or an accelerating photoperiod (short day of LD8:16 from May 10) combined with either warmed (ca 2°C above ambient; 8L-warm) or cooled water (ca 2°C below ambient; 8L-cold) from May to September. Monthly samples were collected from 10 females/group for determination of transcript levels of pituitary gonadotropin subunits (fshb and lhb) and ovarian gonadotropin receptors (fshr and lhr), plasma sex steroids (testosterone: T and estradiol-17ß: E2), gonadosomatic index (GSI) and oocyte size. Short day in combination with either warmed or cooled water induced an earlier increase in pituitary fshb and lhb levels compared with NL-amb controls, and advanced ovarian growth and the seasonal profiles of T, E2. By contrast only minor effects were seen of the photothermal treatments on ovarian fshr and lhr. The 8L-cold had earlier increase in fshb, lhb and E2, but similar oocyte and gonadal growth as 8L-warm, suggesting that the 8L-cold group tried to compensate for the lower water temperature during the period of rapid gonadal growth by increasing fshb and E2 production. Both the 8L-warm and 8L-cold groups showed incomplete ovulation in a proportion of the females, possibly due to the photoperiod advancement resulting in earlier readiness of spawning occurring at a higher ambient temperature, or due to some reproductive dysfunction caused by photothermal interference with normal neuroendocrine regulation of oocyte development and maturation.

Full-term newborn after repeated ovarian tissue transplants in a patient treated for Ewing sarcoma by sterilizing pelvic irradiation and chemotherapy.

We report the first successful transplantation of cryopreserved ovarian cortical tissue into heavily irradiated tissues in a patient who had received sterilizing pelvic radiotherapy (54 Gy) and 40 weeks of intensive high-dose chemotherapy for the treatment of Ewing's sarcoma 14 years earlier. Repeated transplantation procedures were required to obtain fully functional follicular development. Enlargement of the transplants over time and increase of the size of the uterus were demonstrated on sequential ultrasonographic examinations. Eggs of good quality that could be fertilized in vitro were obtained only after a substantial incremental increase of the amount of ovarian tissue transplanted. Single embryo replacement resulted in a normal pregnancy and the birth of a healthy child by cesarean section at full-term. No neonatal or maternal postoperative complications occurred. Women facing high-dose pelvic radiotherapy should not be systematically excluded from fertility preservation options, as is currently the trend.

The role of ionizing radiaton on ovulation rate and oocyte morphology in mouse.

We investigated the effects of ionizing radiation on maturation ability and radiosensitivity of oocytes enclosed in preantral and antral follicles. Balb/c female mice received total body single dose gamma radiation (7.2 Gy) at the diestrous to proestrous transition period. In the first experiment, spontaneously ovulated oocytes were collected from irradiated animals. In the second experiment, irradiated animals were allowed to superovulate to assess the ovarian function. The spontaneous ovulation rate of the follicles exposed at antral stage was significantly lower than the sham-irradiated mice (p < 0.01), and most of the oocytes were found at the metaphase I stage. Oocyte morphology and the ovulation rate of the follicles exposed at preantral stage were similar to the sham-irradiated group. Minimal morphological abnormalities were observed in the oocytes and the polar body as well. The superovulation response of all the irradiated animals was lower than the respective control animals. The superovulation rate was significantly lower in the first ovulation after irradiation (p < 0.01). In conclusion, our findings indicate that total body gamma irradiation, on a basis of estrous cycle stages, leads to ovulation failure in the antral stage while causes abnormal oocyte morphology in the preantral stage follicles in mice.

Effects of spring season solar drying process on sanitation indicators in sewage sludge and potential as a method for fertilizer production.

The agricultural use of sewage sludge is possible on condition of maintaining microbiological and parasitological standards, and one of the most modern methods improving its sanitary state is solar drying. In the presented study, the effect of this process on the elimination of indicator microorganisms (Escherichia coli, Salmonella Senftenberg W775, Enterococcus spp.) and eggs of Ascaris suum introduced into the biomass of sludge was examined. The experiment was carried out in the spring period with a maximal temperature of 18 °C inside the drying plant. Bacteria and parasite eggs were introduced into special carriers (cylinders filled with sewage sludge) and placed at selected points of the drier. The carriers were removed every 7 days and subject to a research procedure in order to estimate the number of bacteria and percentage of live eggs of Ascaris suum. Sanitization of the material was not obtained, since after 28 days of the process the final product contained a large concentration of Enterococcus spp. and S. Senftenberg W775 (10(5) -10(9) MPNg(-1)). Only the number of E. coli decreased by 6 log. During the process, the fastest decrease in the number of bacteria was observed in E. coli (ca 0.2 log/day), slower in enterococci (0.02-0.081 log/day), and the slowest in bacilli of the genus Salmonella (0.011-0.061 log/day). Sludge after drying also still contained 57-66% of live eggs of A. suum. The study proved that the solar drying of sludge in the spring period results in a product which poses a hazard for public and animal health and environmental sustainability, and should not be used as a fertilizer.

Ovarian response to controlled ovarian hyperstimulation in cancer patients is diminished even before oncological treatment.

OBJECTIVE: To evaluate the results of controlled ovarian stimulation before chemotherapy for oocyte vitrification to preserve fertility in women diagnosed with cancer and compare them with a historical control group. DESIGN: A retrospective, multicenter, observational study performed between March 2007 and January 2011. SETTING: University-affiliated infertility clinics. PATIENT(S): Of 272 patients affected by cancer in our Fertility Preservation Program, 223 women underwent a stimulated cycle for oocyte vitrification according to our protocols before cancer treatment. Their results were compared with a historical control group of 98 patients diagnosed with male factor infertility who were stimulated for a conventional IVF cycle. INTERVENTION(S): Controlled ovarian stimulation and oocyte retrieval. MAIN OUTCOME MEASURE(S): Days of stimulation, total dose of gonadotropins, estrogen levels, and number of oocytes retrieved and vitrified. RESULT(S): No differences were found in days of stimulation, but significant differences in E(2) levels and the number of retrieved oocytes were measured, especially in the hormone-dependent cancer group. CONCLUSION(S): Patients with hormone-dependent cancer had a weaker response to controlled ovarian stimulation compared with patients with non-hormone-dependent cancer. Whether the oncological disease already affects the ovaries before chemo-/radiotherapy remains to be elucidated.

Effects of nutritional cues on the duration of the winter anovulatory phase and on associated hormone levels in adult female Welsh pony horses (Equus caballus).

BACKGROUND: Mares have an annual reproductive rhythm, with a phase of inactivity in midwinter. The aim of this study was to determine the impact of food restriction on physiological and metabolic hallmarks of this rhythm. METHODS: Over three successive years, 3 groups of 10 mares were kept under natural photoperiod. A 'well-fed' group was fed to maintain the mares in good body condition; a 'restricted' group received a diet calculated to keep the mares thin and a 'variable' group was fed during some periods like the 'restricted' group and during some other periods like the 'well-fed' group, with the aim of mimicking the natural seasonal variation of pasture availability, but a few months in advance of this natural rhythm. RESULTS: Winter ovarian inactivity always occurred and was long in the restricted group. In contrast, in the 'well-fed' group, 40% of mares showed this inactivity, which was shorter than in the other groups. Re-feeding the 'variable' group in autumn and winter did not advance the first ovulation in spring, compared with the 'restricted' group. Measurements of glucose and insulin concentrations in mares from the 'restricted' group during two 24 h periods of blood sampling, revealed no post-prandial peaks. For GH (Growth hormone), IGF-1 and leptin levels, large differences were found between the 'well-fed' group and the other groups. The glucose, insulin, GH and leptin levels but not melatonin level are highly correlated with the duration of ovulatory activity. CONCLUSIONS: The annual rhythm driven by melatonin secretion is only responsible for the timing of the breeding season. The occurrence and length of winter ovarian inactivity is defined by metabolic hormones.

Ovulatory activity of female goats adapted to the subtropics is responsive to photoperiod.

The objective of this study was to determine whether reproductive seasonality of local well-fed female goats from subtropical Mexico (26 degrees N) can be controlled by photoperiod. The control group (n=12) remained in an open pen under natural photoperiod variations. The two experimental groups (n=8 each) were placed in light-proof rooms and exposed for 2 yr to alternations of 3 months of long days and 3 months of short days. One group was first exposed to long days, Group 1, and the other one to short days, Group 2. Blood samples were obtained twice a week to determine ovulation status by progesterone plasma concentrations. Goats from the control group displayed a seasonality of ovulations. The mean (+/-SEM) dates of the onset and end of the ovulations were September 10+/-5 d and February 16+/-4 d, respectively. In contrast, in both experimental groups, ovulations were modified (P<0.05) by the photoperiodic treatments such that ovulations started and ended during short and long days, respectively. In Groups 1 and 2, ovulations started 67+/-2 and 66+/-2 d (P>0.05), respectively, after the transfer from long to short days. In contrast, the timing of the cessation of ovulations after the transfer from short to long days differed (P<0.05) between groups (19+/-3 and 31+/-3 d for Groups 1 and 2, respectively). Local female goats from subtropical Mexico are, therefore, sensitive to photoperiodic changes that they are exposed to at this latitude and this environmental cue may control the timing of the breeding season in natural conditions.

Reproductive activity of Pelibuey and Suffolk ewes at 19 degrees north latitude.

Under photoperiod regulated conditions, some Pelibuey ewes appear to express estrous cycles continuously throughout the year. Ovulatory activity of Suffolk ewes has not being determined at 19 degrees N. The aim of the present study was to describe the annual reproductive cycle of Pelibuey and Suffolk ewes under natural conditions at 19 degrees N. Ten adult ewes of each breed were maintained under natural environmental conditions, on a constant plane of nutrition and isolated from rams. The ovulatory activity of both breeds was monitored over 2 years (2003 and 2004) by quantifying progesterone concentrations in blood samples taken twice a week. For both breeds, the proportion of ovulating ewes per month and year was analyzed by chi(2). The duration of anestrus was compared between breeds and years by analysis of variance. Correlation of ambient temperature and weight changes with the ovulatory activity was evaluated by regression procedure. Ovulatory activity was different (P<0.05) between breeds from February to July. Pelibuey sheep (60%, 6 of 10) ovulated continuously during the entire study; one animal was eliminated in the second year, due to ovarian follicular cysts and three ewes had anestrous periods from 65+/-46 to 70+/-36 days. Suffolk ewes displayed reproductive seasonality similar to that observed in sheep of the same breed in latitudes greater than 35 degrees . Within breed, reproductive behavior of sheep was similar between years, with the only difference (P<0.05) observed in February for Pelibuey and in July for Suffolk relative to other months. Ambient temperature was not correlated with ovulatory activity. The body weight of Pelibuey ewes was constant during the entire study and was not correlated with ovulatory activity. There was, however, an increase on the body weight of Suffolk ewes, but this variation was not related to ovulatory activity. These findings lead to the conclusion that at 19 degrees N, a high proportion of Pelibuey ewes is capable of ovulating throughout the year, whereas Suffolk ewes undergo seasonal anestrous periods similar to those observed in individuals of the same breed at latitudes greater than 35 degrees . Identification of continuous ovulatory Pelibuey ewes may be the first step for developing a selection program against seasonality to increase lambing frequency.

Germ cell loss induced by 12C6+ ion irradiation in young female mice.

The ovaries of Kun-Ming strain mice (3 weeks) were irradiated with different doses of 12C6+ ion in the Bragg peak or the plateau region. At 10th day after irradiation, ovarian and uterine weights were measured; normal and atretic (identified with the oocyte to be degenerating or absent) primordial, primary and preantral follicles were identified in the largest cross-section of each ovary. Percentage (%) of normal follicles of each developmental stage of oogenesis was calculated. The data showed that compared to controls, there was a dose-related decrease in percentage of normal follicles in each developmental stage. And the weights of ovary and uterus were also reduced with doses of irradiation. Moreover, these effects were much more significant in the Bragg peak region and the region close to the Bragg peak than in the beam's entrance (the plateau region). Radiosensitivity varied in different follicle maturation stages. Primordial follicles, which are thought to be extremely sensitive to ionizing irradiation, were reduced by 86.6%, while primary and preantral follicles reduced only by 72.5% and 61.8% respectively, by exposure with 6 Gy of 12C6+ ion in the Bragg peak region and the region close to the Bragg peak. The data suggested that due to their optimal depth-dose distribution in the Bragg peak region, heavy ions are ones of the best particles for radiotherapy of tumors located next of vital organs or/and surrounded by normal tissues, especially radiosensitive tissues such as gonads.

Human oocyte radiosensitivity.

The teratogenic effects of ionizing radiation are well documented, but less is known about the radiosensitivity of the human oocyte. Women of childbearing age are understandably concerned about diagnostic radiology examinations--their risk/benefit ratio and their impact on the ability to conceive. Current research and evidence gathered from studies of the long-term effects of intra-abdominal therapeutic radiation shed light on this subject. With respect to this issue, radiographers can now give more practical advice to their nongravid female patients.

Spontaneous conception in a young woman who had ovarian cortical tissue cryopreserved before chemotherapy and radiotherapy for a Ewing's sarcoma of the pelvis: case report.

We report the case of a 14 year old girl who presented with a non-metastatic Ewing's sarcoma involving her superior pubic ramus. She received 14 courses of alkylating agent-based chemotherapy and direct radiation to her hemi-pelvis (55 Gy) and is alive and disease-free 8 years later. Multiple biopsies of ovarian cortical tissue were cryopreserved, with her written consent, before treatment began. Ovarian failure was confirmed on completion of treatment with cessation of menses and persistently elevated serum gonadotrophin and low estradiol levels on repeated measurement over 2 years. HRT was initiated. Irregular vaginal bleeding occurred due to radiation vaginitis. Reimplantation of ovarian cortical tissue was considered at 19 years as fertility was desired, but the decision deferred. A spontaneous conception occurred 1 year later and a healthy boy (birthweight 2.9 kg, 3rd-10th centile) was delivered at term by elective Caesarean section. This is the first case of a spontaneous conception occurring in a young woman with documented ovarian failure in whom ovarian cortical tissue had been cryopreserved. Clinicians should be aware of the possibility of spontaneous conception despite confirmed ovarian failure in young women successfully treated for cancer.

Effects of mobile phone radiation on reproduction and development in Drosophila melanogaster.

In this report we examined the effects of a discontinuous radio frequency (RF) signal produced by a GSM multiband mobile phone (900/1,900 MHz; SAR approximately 1.4 W/kg) on Drosophila melanogaster, during the 10-day developmental period from egg laying through pupation. As found earlier with low frequency exposures, the non-thermal radiation from the GSM mobile phone increased numbers of offspring, elevated hsp70 levels, increased serum response element (SRE) DNA-binding and induced the phosphorylation of the nuclear transcription factor, ELK-1. The rapid induction of hsp70 within minutes, by a non-thermal stress, together with identified components of signal transduction pathways, provide sensitive and reliable biomarkers that could serve as the basis for realistic mobile phone safety guidelines.

Laparoscopic ovarian diathermy in women with polycystic ovarian syndrome: a retrospective study on the influence of the amount of energy used on the outcome.

BACKGROUND: Currently, there is an uncertainty about the optimum number of punctures to be applied at laparoscopic ovarian diathermy (LOD). This retrospective study was undertaken to investigate the dose-response relationship of LOD. METHODS: The hospital records of 161 women with polycystic ovarian syndrome who underwent LOD were reviewed and the clinical data before and after LOD were documented. Subjects were divided into six groups according to the number of punctures made in their ovaries as follows: group 1, two punctures per ovary; group 2, three punctures; group 3, four punctures; group 4, five punctures; group 5, six punctures and group 6, seven to 10 punctures. Contingency table analysis and analysis of variance were used to compare the outcomes of the different groups. RESULTS: The rates of ovulation, conception and restoration of menstrual regularity after LOD were significantly lower in group 1 compared with other groups. There were no significant differences in the success rates between the other groups. CONCLUSION: Two punctures per ovary are associated with poor results. Three punctures per ovary seem to represent the plateau dose. The application of seven or more punctures per ovary may result in excessive destruction to the ovary without any improvement of the results and should therefore be discouraged.

[Problems with radiation protection for adjuvant radiotherapy of thyroid cancer]. / Problemy ochrony radiologicznej w izotopowym leczeniu uzupelniajacym raka tarczycy.

Ablative treatment of thyroid cancer with 131I is associated with the exposure to ionizing radiation of not only the target cells but also other cells and organs in the patient's body. Thermoluminescent dosimetry showed that the mean dose equivalent in ovaries equaled to several dozen mSv when the mean therapeutic dose was 2.8 GBq of 131I. In the treated young women the performed ovulation tests revealed no dysfunction of the ovaries. The highest dose equivalent in the clinic staff would not exceed 20 mSv per year. Accordingly, the dose equivalents to the public would not exceed the threshold limit of 1 mSv per year. In order to fulfill the requirements of regulations for disposal of radioactive waste and the necessity to store the waste for as long as 80 days it is obligatory to utilize setting tanks of very large capacity.