Dermal repeated dose toxicity study of the anti-breast cancer drug Formestane cream in Bama minipig.

Formestane (4-OHA) has been proven to be highly effective with high systemic tolerability in treating ER+ breast cancer. However, its intramuscular administration and associated side effects make it unsuitable for adjuvant treatment, leading to its withdrawal from the market. In contrast, Formestane cream may offer a solution by providing a more convenient route of administration and retaining its tumor-shrinking effects. This suggests that 4-OHA cream could have promising clinical applications. However, before clinical application, it is necessary to evaluate the potential toxicity of the cream in animals. This study evaluated the toxicity of 4-OHA cream on female Bama minipigs in vivo by analyzing hematology, biochemistry, and histopathology. The results showed that there was no significant difference between the cream-treated group and the control normal group for each parameter analyzed, indicating that 4-OHA cream was non-dermal toxic to minipigs. This finding provides a basis for the safe clinical use of the cream.

Short-term aromatase inhibition induces prostatic alterations in adult wistar rat: A biochemical, histopathological and immunohistochemical study.

PURPOSE: This study aimed to evaluate the effects of estrogen reduction on amyloid deposition, some lipid metabolism and oxidative stress markers, PSA-like production and p63 expression in the prostate of the adult rat. METHODS: Aromatase inhibitor: Formestane (4-OHA), was administrated to male rats, at a dose of 0.1 mg/kg b.w./day, for 10 days. The control group (CONT) received the same volume of placebo injection (NaCl 0.9%). RESULTS: 4-OHA treatment induced a significant accumulation of intraprostatic cholesterol (138.90 ±â€¯17.64 vs 85.12 ±â€¯2.87, p = 0.01); against an insignificant diminution of malondialdehyde (412.6 ±â€¯54.35 vs 842.70 ±â€¯336.50, p > 0.05) and glutathione (2.40 ±â€¯0.23 vs 3.65 ±â€¯0.88, p > 0.05). This was associated with a significant decrease of nitric oxide (31.76 ±â€¯7.07 vs 179.40 ±â€¯58.35, p = 0.024). Additionally, 4-OHA significantly increased the intraprostatic production of PSA-like (11.12 ±â€¯2.78 vs 3.91 ±â€¯0.43, p = 0.043). The prostatic histology revealed an amyloid deposition, in all prostatic lobes and a smooth muscle layer growth (p < 0.05); especially significant in the dorsal and lateral lobes. Theses lobes manifested a basal cells proliferation, with a 3-fold increase of p63 expression (p < 0.001). The ventral lobe presented epithelial atrophy (37.80 ±â€¯16.20 vs 167.60 ±â€¯5.16, p < 0.05); with occasional and significant proliferative foci (247.00 ±â€¯9.573 vs 167.60 ±â€¯5.16 p < 0.05). DISCUSSION AND CONCLUSION: Aromatase inhibition, in the adult male rat, alters the prostatic function by reducing nitric oxide availability and inducing amyloid deposition along with limiting the differentiation of basal cells, through a lobe-specific p63-overexpression.

TRPA1 Mediates Aromatase Inhibitor-Evoked Pain by the Aromatase Substrate Androstenedione.

Aromatase inhibitors (AI) induce painful musculoskeletal symptoms (AIMSS), which are dependent upon the pain transducing receptor TRPA1. However, as the AI concentrations required to engage TRPA1 in mice are higher than those found in the plasma of patients, we hypothesized that additional factors may cooperate to induce AIMSS. Here we report that the aromatase substrate androstenedione, unique among several steroid hormones, targeted TRPA1 in peptidergic primary sensory neurons in rodent and human cells expressing the native or recombinant channel. Androstenedione dramatically lowered the concentration of letrozole required to engage TRPA1. Notably, addition of a minimal dose of androstenedione to physiologically ineffective doses of letrozole and oxidative stress byproducts produces AIMSS-like behaviors and neurogenic inflammatory responses in mice. Elevated androstenedione levels cooperated with low letrozole concentrations and inflammatory mediators were sufficient to provoke AIMSS-like behaviors. The generation of such painful conditions by small quantities of simultaneously administered TRPA1 agonists justifies previous failure to identify a precise link between AIs and AIMSS, underscoring the potential of channel antagonists to treat AIMSS. Cancer Res; 76(23); 7024-35. ©2016 AACR.

Anabolic and androgenic activity of 19-norandrostenedione after oral and subcutaneous administration--analysis of side effects and metabolism.

One of the most frequently misused steroid precursors (prohormones) is 19-norandrostenedione (estr-4-ene-3,17-dione, NOR). Recently we have show that NOR stimulates skeletal muscle growth after s.c. administration in a highly selective manner but exhibits only weak androgenic activity in rats. Because most abusers take NOR orally, the aim of this study was to compare the anabolic and androgenic potency of NOR between s.c. and oral application. Orchiectomised rats were treated with NOR either s.c. (1 mg/kg BW/day) or orally (0.1, 1 and 10 mg/kg BW/day). The tissue weights of the levator ani, the seminal vesicle and the prostate were analysed to determine the anabolic and androgenic activity. Heart and liver wet weights were examined to identify side effects. Serum concentrations of NOR and its metabolite nandrolone (NT) were determined. GCMC analysis revealed that free and glucuronidated NOR and NT were detectable in the serum after oral and s.c. administration and that NOR was converted to NT in comparable amounts independent of the route of administration. In agreement to our previous study s.c. application of NOR stimulates skeletal muscle growth but has only weak androgenic effects. In contrast, after oral administration of NOR neither stimulation of the prostate nor the levator ani could be observed in the doses administered in this study. Interestingly, and in contrast to s.c. treatment, oral administration of NOR resulted in a dose-dependent decrease of body weight. In summary, oral administration of NOR, at least in the rat, seems to be a very ineffective strategy for stimulating skeletal muscle mass increases but may be associated with side effects.

Supplements and the endocrine system in athletes.

In the world of athletes' nutrition, there are many ethical concerns, because there is the suspicion that in practice, large doses of supplements in athletes are not taken for nutritional purposes. It is beyond the scope of this article to highlight the possible roles of supplements or methods of supplementation in the improvement of athletic performance in elite athletes. Instead, the author briefly reviews some of the substances taken by athletes, with particular attention to their mechanisms of action and the pathways involved. Very often, the effects of many supplements are hormone-related, or supplements influence hormone secretion. Examples of possible links between "supplements or ergogenic compounds" and the endocrine/metabolic system are addressed.

Testosterone precursors: use and abuse in pediatric athletes.

The dietary supplements androstenedione, dehydroepiandrosterone, and androstenediol are precursors in the endogenous production of testosterone. The efficacy and safety of these prohormones are not well established but are promoted to have the same androgenic effects on building muscle mass and strength as anabolic-androgenic steroids. Studies have demonstrated repeatedly that acute and long-term administration of these oral testosterone precursors does not effectively increase serum testosterone levels and fails to produce any significant changes in lean body mass, muscle strength, or performance improvement compared with placebo. The Anabolic Steroid Control Act of 2004 lists androstenedione as a schedule III controlled substance, and it is regulated by the U.S. Food and Drug Administration. Testosterone precursors are banned by most major sports organizations.

Concentrações hormonais e desenvolvimento folicular de vacas leiteiras em hipertermia sazonal e aguda / Hormone concentration and follicular development in dairy cows under seasonal and acute hyperthermia

Avaliaram-se as concentrações hormonais e os parâmetros de desenvolvimento folicular de vacas leiteiras expostas ao calor sazonal e agudo. Dividiram-se os animais em quatro grupos: verão (n=5), outono (n=5), inverno com hipertermia aguda (grupo câmara climática, (CC), n=5) e inverno (n=9). Os animais foram abatidos no sétimo dia após a ovulação, e os parâmetros de desenvolvimento folicular avaliados. O líquido folicular do maior folículo foi aspirado e armazenado para posterior análise de hormônios esteróides e inibina. O número de células da granulosa vivas no verão e no outono foi 40 e 45 por cento respectivamente, menor que no inverno (P<0,05). A concentração de estradiol (E2) no inverno foi 62 por cento maior que no outono (P<0,05) e 34 por cento superior ao grupo verão (P<0,06). Houve um aumento na quantidade de androstenediona no verão em relação aos grupos inverno (P<0,08) e outono (P<0,05). A concentração de inibina foi maior no inverno do que no verão e CC (P<0,05). A exposição ao calor sazonal e agudo modificou os parâmetros de desenvolvimento do folículo e as concentrações hormonais no líquido folicular, podendo explicar em parte a queda nas taxas de concepção no verão.

The present study evaluated the seasonal and acute heat stress on follicular development and steroid and inhibin concentrations in follicular fluid, in bovine dominant follicle. Cows were distributed into four treatments: summer (n=5), autumn (n=5), animals heat stressed during the winter (n=5) and winter (n=9). On day 7 of the estrous cycle, animals were slaughtered and parameters related to follicle development were evaluated. The follicular fluid (FF) was aspirated and stored for further hormonal analysis. During the summer, the number of viable granulosa cells was 40 percent lower than during the winter, and there was a 45 percent decrease in this parameter during the autumn (P<0.05). In the winter, estradiol concentration was 62 percent higher than during the autumn (P<0.05) and 42 percent higher than during the summer (P<0.06). There was an increase in androstenedione concentration in summer group, when compared to winter (P<0.08) and autumn (P<0.05) groups. Inhibin concentration was higher in winter groups than summer and winter heat stressed groups (P<0.05). Seasonal and acute heat stress altered developmental parameters in dominant follicle and hormonal concentration in follicular fluid, those effects can partially explain the decrease in conception rates during summer.

Testosterone prohormone supplements.

Testosterone prohormones such as androstenedione, androstenediol, and dehydroepiandrosterone (DHEA) have been heavily marketed as testosterone-enhancing and muscle-building nutritional supplements for the past decade. Concerns over the safety of prohormone supplement use prompted the United States Food and Drug Administration to call for a ban on androstenedione sales, and Congress passed the Anabolic Steroid Control Act of 2004, which classifies androstenedione and 17 other steroids as controlled substances. As of January 2005, these substances cannot be sold without prescription. Here, we summarize the current scientific knowledge regarding the efficacy and safety of prohormone supplementation in humans. We focus primarily on androstenedione, but we also discuss DHEA, androstenediol, 19-nor androstenedione, and 19-nor androstenediol supplements. Contrary to marketing claims, research to date indicates that the use of prohormone nutritional supplements (DHEA, androstenedione, androstenediol, and other steroid hormone supplements) does not produce either anabolic or ergogenic effects in men. Moreover, the use of prohormone nutritional supplements may raise the risk for negative health consequences.

Oral androstenedione-induced impotence and severe oligospermia.

OBJECTIVE: To report a case of androstenedione (A)-induced impotence and severe oligospermia. DESIGN: Case report. SETTING: Multispecialty tertiary referral center. PATIENT(S): A 29-year-old married male recreational bodybuilder using the legal dietary supplement A in an attempt to enhance athletic performance. INTERVENTION(S): Immediate cessation of oral A, Depo-T regimen with a taper, administered during 3 months, to alleviate loss of libido and impotence. MAIN OUTCOME MEASURE(S): Resolution of symptoms and normalization of semen parameters. RESULT(S): Resolution of symptoms and normalization of semen parameters at 6 months, and successful intrauterine pregnancy achieved 1 year after initial presentation. CONCLUSION(S): Oral A has a previously unrecognized side effect, which is the potential to suppress the hypothalamic pituitary gonadal axis.

The estrogen suppression after sequential treatment with formestane in advanced breast cancer patients.

In postmenopausal patients, estrogens have an important role in breast cancer growth and aromatase inhibitors (AI) suppress the aromatase enzyme system which converts androgens into estrogens. The aim of this study was to evaluate the effect on estrogen suppression of formestane 250 mg i.m. fortnightly, given immediately after the failure of a previous treatment with non-steroidal AI. Twenty-two advanced breast cancer patients progressing on letrozole, anastrozole and aminoglutethimide entered the study. At the beginning of the study, the serum estrogen levels were suppressed by the previous treatment with non-steroidal AI, and the following treatment with formestane moderately maintained this suppression; in four patients serum estrogen levels increased fivefold after 10 weeks. Neither complete nor partial responses were observed; 11 patients (50%) showed a stable disease lasting > or = 6 months, and the median time to progression was 6 months (range 3-9 months). No correlation was observed between clinical responses and serum estrogen suppression. Tolerability was satisfactory, and no patient withdrew from the study due to adverse events. In conclusion, formestane has demonstrated a moderate activity in estrogen suppression, and there is evidence that, at the failure of a previous treatment with non-steroidal AI, the sequential use of steroidal AI is feasible. This approach can be used in clinical practice in order to offer a disease control with a satisfactory quality of life.

Morphometric analysis of the hormonal effect on tissue-implant response associated with TCP bioceramic implants.

Recent studies in our laboratory have demonstrated the need for an experiment comparing the effect of steroid hormones on the cellular composition of the fibrous tissue capsules surrounding tricalcium phosphate (TCP) bioceramics. The effect of steroid hormones on inflammatory cells has been widely reported in the scientific literature. The purpose of this research is to contribute to the development of a long-term ceramic drug delivery system with human and veterinary applications. This study contributes information on the composition of fibrous tissue capsules surrounding hormone loaded TCP bioceramic implants at 90 days post-implantation. The data obtained in this study was gathered using protocols developed from ImagePro software. Sixteen animals in four experimental groups were implanted with one TCP bioceramic each. Group I animals were implanted with a sham TCP ceramic not containing a steroid hormone (control group). Group II animals received the testosterone loaded TCP ceramic. Group III animals were implanted with the dihydrotestosterone loaded ceramic. Group IV animals received the androstenedione ceramic. At 90 days post-implantation, the animals were euthanized. The implants and fibrous tissue capsules surrounding them were then extracted. The fibrous tissue capsules were evaluated microscopically using ImagePro software following routine II&E staining, modified Papanicolau, and Masson's trichrome. The research variables comparing the hormonal effects on fibrous tissue composition obtained by digital analysis were as follows: thickness of the fibrous tissue, quantification of macrophages, neutrophils, fibroblasts, vascularity, and area occupied by collagen. The results of this study support several conclusions. All three of the hormones in this study, particularly androstenedione, have significant affects on the thickness and cellular composition of the fibrous tissue capsules when compared to the control group. Digital analysis software enables more accurate and reproducible results to be obtained when specific procedures are followed.

Immunohistochemical detection of cytokine expression in tissue-implant response associated with TCP bioceramic implants loaded with steroid hormones.

Recent studies in our laboratory have demonstrated the need for an experiment comparing cytokine expression in fibrous tissue capsules surrounding tricalcium phosphate (TCP) bioceramics loaded with androstenedione, dihydrotestosterone, and testosterone to that of a control to further elucidate the mechanisms involved in the tissue-implant response. The effect of steroid hormones on inflammatory cells has been widely reported in the scientific literature. The purpose of this research is to contribute to the development of a long-term ceramic drug delivery system with human and veterinary applications. This study contributes information on the presence of cytokines in fibrous tissue capsules surrounding hormone loaded TCP bioceramic implants at 90 days post-implantation. Sixteen animals in four experimental groups were implanted with one TCP bioceramic each. Group I animals were implanted with a sham TCP ceramic not containing a steroid hormone (control group). Group II animals received the testosterone loaded TCP ceramic. Group III animals were implanted with the dihydrotestosterone loaded ceramic. Group IV animals received the androstenedione ceramic. At 90 days post-implantation, the animals were euthanized. The implants and fibrous tissue capsules surrounding them were then extracted. The fibrous tissue capsules were evaluated microscopically following routine H&E staining and IHC staining of antibodies to IL-1 beta, Il-2, IL-6, TNF infinity, and cell specific markers for CD-4 and CD14 positive cells. The results of this study indicate that these hormones, particularly androstenedione, limit the expression of cytokines and greatly affect the cellular composition of the tissue-implant response.

Oral andro-related prohormone supplementation: do the potential risks outweigh the benefits?

Androstenedione, 4-androstenediol, 5-androstenediol, 19-norandrostenediol and 19-norandrostenedione are commonly referred to as "Andro" prohormones. Over the last few years, supplementation using these prohormones has been aggressively marketed to the general public. Supplement manufacturers often claim that Andro use improves serum testosterone concentrations, increases muscular strength and muscle mass, helps to reduce body fatness, enhances mood, and improves sexual performance. However, to date, most studies contradict these claims. In contrast, several studies using oral Andro related prohormones show that Andro use can abnormally elevate estrogen related hormones as well as alterations in hormonal markers (i.e., abnormal elevations in serum estrogen) thought to increase a person's risk for developing prostate or pancreatic cancers. In addition, most studies also indicate that significant declines in high-density lipoproteins occur leading to an increased cardiovascular disease risk. Thus, to date, the current research base suggests that Andro prohormone use does not support manufacturer claims. But it does suggest there should be strong concerns regarding long-term oral Andro prohormone use, especially regarding its effects on blood lipids and estrogen hormone profiles.

[Clinical trial of lentaron for postmenopausal patients with advanced breast cancer].

OBJECTIVE: To evaluate the efficacy and adverse effects of lentaron for postmenopausal patients with recurrent and metastatic breast cancer. METHODS: Thirty-four patients with recurrent and metastatic breast cancer received 250 mg lentaron by intramuscular injection every 2 weeks for at least one month. RESULTS: In 34 patients who were evaluable for efficacy and toxicity, the complete response rate (CR), partial response rate (PR), disease stabilization rate (SD) and progressive disease rate (PD) were 0%, 14.7%, 58.8% and 26.5%. The clinical benefit rate (CR + PR + SD >/= 6 months) was 50.0%. (17/34) with 12 patients (35.3%) having SD for at least 6 months. The response rates for bone, soft tissue and visceral metastasis were 28.6% (3/14), 13.6% (3/22) and 5.3% (1/19), respectively. There were no severe adverse effects in the treatment bylentaron. CONCLUSION: Lentaron is a well tolerated agent with reasonable efficacy but low toxicity for postmenopausal patients with recurrent and metastatic breast cancer.

Evaluation of androstenedione as an androgenic component of river water downstream of a pulp and paper mill effluent.

This study evaluates a recent report indicating that androstenedione (4-androsten-3, 17-dione) contributes to the androgenicity of water downstream of a pulp and paper mill discharge on the Fenholloway River (FL, USA). Extraction and concentration of Fenholloway water with C18 solid-phase extraction columns followed by reverse-phase high-pressure liquid chromatography resulted in clearly defined fractions with in vitro androgenic activity in CV-1 cells that had been transiently cotransfected with human androgen receptor and reporter gene constructs. However, we were unable to detect androstenedione in the active fractions by gas chromatography/mass spectrometry. Mass spectrometry analyses of deionized and Fenholloway River water samples that had been spiked with androstenedione, then extracted and fractionated, revealed that the androgen was found only in inactive fractions. We conclude that, although androstenedione was present at easily detectable concentrations in the river water (> 100 ng/L), this compound is not associated with androgenic activity of water from the site.