Restoration of fertility in light-deprived female hamsters by chronic melatonin treatment

Summary Blinding young adult female hamsters was followed by functional involution of the ovaries and uteri and by the cessation of cyclic vaginal phenomena. Light deprivation was also accompanied by elevated plasma and pituitary levels of luteinizing hormone and depressed levels of prolactin in both the blood and the pituitary gland. Only one of 15 blinded hamsters became pregnant when they were exposed to fertile males for 30 days. Both pinealectomy or chronic melatonin treatment (1 mg melatonin implanted subcutaneously per week in beeswax) prevented the changes in the reproductive organs and in pituitary hormone levels attendant on light-deprivation. Both treatment also returned vaginal cycles to normal and restored plasma prolactin titers. Unlike hamsters that were blinded only, light deprived hamsters that were either pinealectomized or melatonin treated were capable of reproducing when they were caged with fertile males. The reproductive capability (i.e., percent of animals that become pregnant and the sizes of their litters) of these animals was equivalent to that of the untreated control hamsters. This is the first report that chronic melatonin treatment restores fertility in blinded female hamsters.Supported by Grant GB-43233X from the National Science Foundation     

Restoration of fertility with gonadotropin and bromocriptine in a hypogonadal man after removal of macroprolactinoma

A case of male hypogonadism after removal of macroprolactinoma was successfully treated with gonadotropin. A 35-year-old man treated surgically for pituitary adenoma had elevated plasma prolactin and impaired pituitary function after the operation. He was on replacement of hydrocortisone, levothyroxine and testosterone depot along with bromocriptine. Normal plasma testosterone levels were achieved with HCG, 3,000 IU three times a week. The addition of 75 IU of FSH daily restored spermatogenesis and the sperm count reached the fertile range at the 11th month. Doses of HCG and FSH were cut in half at the 10th month without affecting the plasma testosterone levels. His wife was impregnated at the 12th month and gave birth to a normal baby girl.     

Preservation of fertility in females treated for cancer

Advancements of diagnosis and treatment have substantially improved cancer survival rates in the last few decades. The increasing number of survivors focuses attention on long-term effects caused by cancer treatment and its impact on quality of life. Ovarian failure is one of the major sequelae of cytotoxic chemotherapy and/or radiotherapy in female children and reproductive-age women. Oncologists should address the patients about fertility preservation options before therapy. Embryo cryopreservation is the only well-established method for females in preserving fertility; however other strategies including ovarian suppression, ovarian transposition and cryopreservation of oocytes and ovarian tissue are still experimental. Patients need advice and to know which are the most practical options for them. This article reviews the available fertility preservation methods in women, and the related issues including normal physiology of the ovary, effect of anticancer therapy on fertility, role of the oncologist and ethics. We performed a MEDLINE search from 1971 to 2011 in a similar way as Jensen et al. 2011, using the following MeSH terms: antineoplastic agents; ovarian failure; premature; infertility, female; fertility preservation; child and cancer; reproductive technologies, assisted.     

Skin cancer after X-ray treatment for scalp ringworm

Some 2,224 children given X-ray therapy for tinea capitis (ringworm of the scalp) have been followed for up to 50 years to determine cancer incidence, along with a control group of 1,380 tinea capitis patients given only topical medications. The study found a relative risk (RR) of 3.6 (95% confidence interval, 2.3-5.9) for basal cell skin cancer (BCC) of the head and neck among irradiated Caucasians (124 irradiated cases and 21 control cases), in response to a scalp dose of about 4.8 Gy. No melanomas of the head and neck have been seen, and only a few squamous cell carcinomas. About 40% of irradiated cases have had multiple BCCs, for a total of 328 BCCs. Although 25% of both the irradiated and control groups are African-American, only 3 skin cancers have been seen among them, all in the irradiated group, indicating the importance of susceptibility to UV radiation as a cofactor. Light complexion, severe sunburning and North European ancestry were predictive of BCC risk in the irradiated group, but chronic sun exposure was not. Children irradiated at young ages had the highest BCC risk. The RR for BCC risk is approximately constant with time since exposure, suggesting that risk will probably last for a lifetime.     

Restoration of fertility by germ cell transplantation requires effective recipient preparation

Spermatogonial transplantation provides access to the mammalian germline and has been used in experimental animal models to study stem cell/niche biology and germline development, to restore fertility, and to produce transgenic models. The potential to manipulate and/or transplant the germline has numerous practical applications that transcend species boundaries. To make the transplantation technology more broadly accessible, it is necessary to develop practical recipient preparation protocols. In the current study, mouse recipients for spermatogonial transplantation were prepared by treating pregnant females with the chemotherapeutic agent busulfan at different times during gestation. Donor germ cells were introduced into the testes of male progeny between 5 and 12 days postpartum. Analysis of recipient animals revealed that busulfan treatment of pregnant females on 12.5 days postcoitum was the most effective; male progeny transplanted with donor germ cells became fertile and passed the donor genotype to 25% of progeny. This approach was effective because 1) the cytoablative treatment reduced (but did not abolish) endogenous spermatogenesis, creating space for colonization by donor stem cells, 2) residual endogenous germ cells contributed to a healthy testicular environment that supported robust donor and recipient spermatogenesis, and 3) fetal busulfan-treated males could be transplanted as pups, which have been established as better recipients than adults. Laboratory mice provide a valuable experimental model for developing the technology that now can be applied and evaluated in other species.     

Restoration of fertility in sterilized Drosophila eggs by transplantation of polar cytoplasm

A technique for transplantation of cytoplasm between Drosophila eggs is described. Polar cytoplasm of newly laid eggs was first made ineffective for the determination of germ cells by UV irradiation. The sterility which results from this UV irradiation could be prevented by the injection of polar cytoplasm, but not by the injection of anterior cytoplasm from unirradiated donor eggs. The results provide the first demonstration of transplantation of agents causing determination in an insect and also provide a bioassay for these agents.Microscopical observations of UV irradiated eggs with and without transplanted polar cytoplasm revealed that UV irradiation delays the migration of cleavage nuclei into the posterior periplasm and prevents cytoplasmic protrusions at the posterior pole from becoming isolated from the periplasm to form pole cells. Transplantation of polar cytoplasm repairs these defects.     

Restoration of endocrine and ovarian function after stopping GnRH antagonist treatment in goats

We have tested if the high number of unfertilized ova and degenerated embryos found in superovulated goats previously treated with GnRH antagonist can be related to a prolongation of gonadotrophin down-regulation and/or alterations in follicular function during the period of administration of the superovulatory treatment, around 4 days after the end of the antagonist treatment. A total of 15 does were treated with intravaginal progestagen sponges and daily injections of 0.5mg of the GnRH antagonist Antarelix for 6 days, while 5 does acted as controls receiving saline. During the antagonist treatment, the mean plasma LH concentration was lower in treated than control goats (0.5 +/- 0.2 versus 0.7 +/- 0.5 ng/ml, P < 0.0005 ); however, the FSH levels remained unaffected (0.8 +/- 0.4 versus 0.8 +/- 0.5 ng/ml). In this period, treated does also showed an increase in the number of small follicles with 2-3 mm in size ( 10.7 +/- 0.7 versus 8.4 +/- 0.6, P < 0.05), and a decrease in both the number of follicles > or =4 mm in size ( 5.0 +/- 0.3 versus 6.8 +/- 0.5, P < 0.005) and the secretion of inhibin A (120.9 +/- 10.7 versus 151.6 +/- 12.6 pg/ml, P < 0.05). After cessation of the antagonist treatment, there was an increase in LH levels in treated goats from the day after the last Antarelix injection (Day 1), so that LH levels were the same as controls on Day 3 (0.6 +/- 0.1 versus 0.6 +/- 0.2 ng/ml). However, there were even greater numbers of small follicles than during the period of antagonist injections (15.4 +/- 0.6 in treated versus 8.9 +/- 0.7 in control, P < 0.0005 ). Moreover, the number of > or =4 mm follicles and the secretion of inhibin A remained lower in treated goats (3.9 +/- 0.3 follicles and 84.4 +/- 7.0 pg/ml versus 5.4 +/- 0.5 follicles, P < 0.05 and 128.9 +/- 14.2 pg/ml, P < 0.05 ). These results indicate that pituitary secretion of gonadotrophins is restored shortly after the end of antagonist treatment, but activity of ovarian follicles is affected.     

Restoration of the bone marrow pluripotent stem cells in AKR mice after arabinosylcytosine treatment

The hypothesis of repression of multipotent stem cells (CFU) by leukemic cells to explain their depletion, previously demonstrated in AKR leukemic mice, was tested. Using arabinosylcytosine to destroy leukemic cells, it was shown that the bone marrow CFU pool was acutely depressed between 2 h and 12 h after treatment. However, 5 to 7 days later, this pool was restored, surpassing the value of the bone marrow pool in normal mice. This seems to indicate that the CFU pool in leukemic mice is potentially capable of proliferating but is repressed by leukemic cells.     

Endogenous hormone response and fertility in dairy heifers after treatment with Lutalyse and GnRH

Fifty-five dairy heifers were given two injections of Lutalyse 11 days apart. Twenty-one of the heifers were also given an injection of GnRH 48 hr after the second Lutalyse injection (Group G). Of the remaining 34 animals, 19 were randomly allotted to be inseminated 12 hr after observed estrus following Lutalyse (Group E), while 15 were inseminated 80 hr after the second Lutalyse injection (Group P). The intervals from second Lutalyse injection to occurrence of both estrus and peak gonadotropin concentrations were variable among animals receiving only Lutalyse. GnRH injections reduced variation (P<.01) in the interval from second Lutalyse injection to occurrence of peak gonadotropin concentrations, but did not improve fertility. Pregnancy rates did not differ (P>.05) among treatment groups. The failure of GnRH administration following Lutalyse to improve pregnancy rates indicates that GnRH administration followed by insemination 12 hr later is not effective in increasing pregnancy rates above those attained in animals inseminated at either 12 hr post estrus or 80 hr after second Lutalyse injection.     

Breast-conservation treatment for early invasive breast cancer: prognostic factors for survival after salvage treatment of local recurrence

The aim of the study was to investigate prognosis of patients who develop an isolated local recurrence (ILR) after conservative surgery (CS) for early-stage invasive breast cancer. Between 1983 and 1987, 415 patients with stage I-II breast cancer were treated with CS. Of these patients, 68 developed an ILR. The mean follow-up time after ILR was 167 months. Cox models taking potential prognostic factors into account were used to estimate the risk of death. On univariate analysis, age (< or =40 vs. >40 years) at first treatment, time to ILR (< or =24 vs. >24 months), type of recurrence (true vs. new primary tumor, NP), and extent of recurrence (operable vs. inoperable) were, but initial tumor stage (pT1 vs. pT2), initial lymph node stage (pN-negative vs. -positive), adjuvant radiotherapy (yes vs. no), type of salvage surgery (wide excision vs. mastectomy), and recurrent tumor grade (1-2 vs. 3) were not significant predictors of post-recurrence survival. On multivariate analysis only time to ILR proved independent predictor of survival (relative risk: 3.2; 95% confidence interval: 1.4-7.3; p = 0.0051), and the age of the patients showed borderline significance (p = 0.0659). The 15-year actuarial rate of cause-specific survival after ILR was 58% for all patients, 60% and 0% for patients with operable or inoperable recurrence, 30% and 71% for patients with age < or =40 or >40 years, 25% and 72% for patients with time to ILR < or =24 or >24 months, 54% and 88% for patients with true recurrence or NP, and 92% for patients with age >40 years with NP, respectively. The rate of second local recurrence after salvage mastectomy or repeated wide excision was 16% and 28%, respectively (p = 0.2265). As a conclusion, many patients with ILR have good prognosis, particularly those with operable recurrence with prolonged time to ILR, or with NP. Salvage mastectomy is not mandatory for all CS patients.     

Restoration of spermatogenesis and male fertility by transplantation of dispersed testicular cells in the chicken

Transplantation of male germ cells into sterilized recipients has been widely used in mammals for conventional breeding and transgenesis purposes. This study presents a workable approach for germ cell transplantation between male chickens. Testicular cells from adult and prepubertal donors were dispersed and transplanted by injection directly into the testes of recipient males sterilized by repeated gamma irradiation. We describe the repopulation of the recipient seminiferous epithelium up to the production of heterologous sperm in about 50% of transplanted males. In comparison to males transplanted with testicular cell preparations from adult donors, in which the first ejaculates with sperm were recovered about 5 wk after transfer, a substantial interval (about 10 wk) was necessary to obtain ejaculates after the transfer of testicular cells from prepubertal donors. However, in both cases, recipient males produced ejaculates capable of fertilizing ova and producing progeny expressing donor genes.     

Superovulatory response, embryo quality and fertility after treatment with different gonadotrophins in native cattle

We studied native Mertolengo cattle to evaluate superovulatory (SOV) treatments, subsequent fertility of donors and pregnancy rate of recovered embryos. In Experiment 1 we compared superovulatory response (SR), embryo quality and plasma progesterone (P4) levels between donors treated with eCG (10 cows and 5 heifers) vs. FSH (pure, FSH-1, n=10 cows and crude, FSH-2, n=10 cows), during progestagenic impregnation. We also compared fertilization rates and embryo quality of bred and inseminated eCG and FSH-1 donors. Significantly more viable embryos were yielded by FSH than by eCG treated donors. Less FSH-1 than FSH-2-treated donors showed SR, but the response was identical in responder donors of both groups. Fertilization rates were significantly higher in bred than in inseminated donors. Plasma P4 levels were only significantly different (higher) between responder and non-responder donors on the day of embryo recovery. Experiment 2 compared FSH treatments (FSH-2, crude, n=11 cows and FSH-3, pure, n=10 cows) started at the midluteal phase. The mean number of viable embryos was significantly higher in FSH-3 than in FSH-2 treated donors. Both FSH treatments exerted a similar luteotrophic effect upon injection. The FSH-2 donors treated during the midluteal phase yielded more ova and showed significantly higher plasma P4 levels at all sampling days than those treated during progestagenic impregnation. The pregnancy rates of recipient cows were 67% and 46% for fresh and frozen-thawed embryos respectively. In Experiment 3, the fertility of donors (n=20) after SOV treatments was compared with that of untreated cows (n=40). Time to conception of donors, after mating with a bull 14 days after embryo recovery, was identical to that of control cows. There was some delay to conception in eCG-treated cows, but the difference was not significant. These preliminary results suggest that response to SOV treatments in Mertolengo cattle might be affected by the type of gonadotrophin and by the treatment protocol. The fertility of a traditional breeding season after SOV treatments was not impaired. Cryopreserved embryo banking can be used to preserve the breed.     

Reconstruction after conservative treatment for breast cancer: cosmetic sequelae classification revisited

Patients with suboptimal results following breast conservative therapy are presenting more frequently to plastic surgeons as a difficult management problem. A three-type "cosmetic sequelae classification" is proposed to evaluate and manage these patients. From February of 1991 to November of 2001, 85 patients were treated for cosmetic sequelae of breast conservative therapy at the Institut Curie. The patients were followed up prospectively for 6 to 132 months (median, 33 months). They were assessed with regard to age, site and stage of tumor, type of initial breast conservative therapy undertaken, corrective operative procedures performed, complications, and cosmetic results. Forty-eight patients (56.5 percent) had type 1 cosmetic sequelae, 33 patients (38.8 percent) had type 2, and four patients (4.7 percent) had type 3. Type 1 was managed by contralateral symmetrizing procedures. Type 2 was the most difficult to manage by means of various procedures. Type 3 required mastectomy and immediate breast reconstruction. Type 1 had 97.6 percent good results compared with 82.7 percent for type 2. Three of the four type 3 patients had good results. This article reaffirms the validity of the cosmetic sequelae classification as a simple, practical guide for breast reconstructive surgeons. It discusses the various choices of reconstructive procedures available, the importance of preventing these cosmetic sequelae, and the role of the plastic surgeon in the planning of conservative treatment of breast cancers.