Sex Hormones, Growth Hormone, and the Cornea.

The growth and maintenance of nearly every tissue in the body is influenced by systemic hormones during embryonic development through puberty and into adulthood. Of the ~130 different hormones expressed in the human body, steroid hormones and peptide hormones are highly abundant in circulation and are known to regulate anabolic processes and wound healing in a tissue-dependent manner. Of interest, differential levels of sex hormones have been associated with ocular pathologies, including dry eye disease and keratoconus. In this review, we discuss key studies that have revealed a role for androgens and estrogens in the cornea with focus on ocular surface homeostasis, wound healing, and stromal thickness. We also review studies of human growth hormone and insulin growth factor-1 in influencing ocular growth and epithelial regeneration. While it is unclear if endogenous hormones contribute to differential corneal wound healing in common animal models, the abundance of evidence suggests that systemic hormone levels, as a function of age, should be considered as an experimental variable in studies of corneal health and disease.

Development of extraction separation technology based on deep eutectic solvent and magnetic nanoparticles for determination of three sex hormones in milk.

A rapid, reliable and eco-friendly method for the determination of three sex hormones in five kinds of milk was developed and validated by combining vortex-assisted liquid-liquid microextraction (VALLME) and magnetic solid-phase extraction (MSPE). Deep eutectic solvents (DESs) such as choline chloride/urea were considered as the extraction solvent in VALLME and multi-walled carbon nanotubes (MMWCNTs) were used as the adsorbent which could adsorb DESs on the surface. The optimum experimental conditions were as follows: amount of MMWCNTs for 10 mg, volume of acetone for 4 mL, no sodium chloride and extraction pH at 7. After the optimization of several main variables, satisfactory sensitivity levels were achieved as low as 1.0-1.3 ng mL-1 and 2.5-4.5 ng mL-1 for the limit of method detections and the limit of method quantitation, respectively. The recoveries of the three hormones in different milk samples were in the range of 80.1%-116.4%. Consequently, this method is suitable for monitoring the trace amount of sex hormones in milk matrices.

Seasonal changes in reproductive anatomy and gonadal hormone concentrations of African penguins (Spheniscus demersus).

Several standard descriptions of the avian male and female reproductive tract have been reported, including effects of age, stage of reproductive maturity and gonadal hormone concentrations. Limited information on penguin reproductive biology and a lack of information on the African penguin (Spheniscus demersus) necessitated a detailed description of salient structural features of this species and provided an opportunity to evaluate seasonal changes in gonadal steroid hormone concentrations. Tissues from 36 males (adults and juveniles) and 29 females (adults and juveniles) were used for macro-anatomical descriptions and histology of the testes and ovaries. In addition, concentrations of gonadal steroid hormones for eight captive African penguins (four females and four males) were quantified during two breeding and one non-breeding season. The testes were asymmetrical, with the right testis having smaller dimensions compared to the left testis. Marked spermatogenic cellular associations and spermatid developmental stages were present in adult testes only during the breeding season. There was variation in the dimensions of the single ovary during follicular development related to the age and breeding status of the females. Testosterone, dihydrotestosterone, and estradiol concentrations fluctuated during the breeding and non-breeding periods, with males and females having similar steroid concentrations. The results from this study confirm that the breeding status in African penguins can be deduced based on testicular and ovarian histological structures. The results from the present study focused on African penguin reproductive biology should be considered in management strategies for the conservation of the species.

Targeted LC-MS/MS analysis of steroid glucuronides in human urine.

Conjugation with glucuronic acid is one of the major metabolic reactions in human steroid hormone catabolism. Recently, increasing interest has been raised concerning the biological roles of steroid glucuronides. We have therefore developed and validated a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous quantification of 15 urinary steroid hormone glucuronides in human urine: androsterone glucuronide (An-G), etiocholanolone glucuronide (Etio-G), epiandrosterone glucuronide (epiAn-G), dihydrotestosterone glucuronide (DHT-G), dehydroepiandrosterone glucuronide (DHEA-G), testosterone glucuronide (T-G), epitestosterone glucuronide (epiT-G), estrone glucuronide (E1-3 G), 17ß-estradiol 17-glucuronide (E2-17 G), 17ß-estradiol 3-glucuronide (E2-3 G), estriol 16-glucuronide (E3-16 G), pregnenolone glucuronide (Preg-G), tetrahydro-11-deoxycorticosterone 3-glucuronide (THDOC-3 G), cortisol 21-glucuronide (F-G) and pregnanediol glucuronide (PD-G). Sample workup included protein precipitation and solid phase extraction. Internal standards were used to correct for the loss of analytes during sample preparation and analysis. The method showed good linearity (R2≥0.99) and recovery ranged from 89.6 % to 113.8 %. Limit of quantification ranged from 1.9 nmol/L for F-G to 21.4 nmol/L for An-G. Intra-day and inter-day accuracy and precision were below 15 % for all quality controls. The method was successfully applied to 67 urine samples from children and adolescents in whom total concentrations of free and conjugated steroids had been previously determined by GC-MS after enzymatic hydrolysis. Free and sulfated steroids were also measured by LC-MS/MS. In general, the sums of the respective glucuronidated, sulfated and free forms of an analyte corresponded well with its total amount determined after enzymatic hydrolysis by GC-MS. Regarding the most prominent steroid metabolites, the total mean levels of androsterone and etiocholanolone showed an increase up to 5820.0 nmol/L and 4017.8 nmol/L in the group of 15-20 year-old children, respectively. Glucuronide conjugates (4374.3 nmol/L and 3588.5 nmol/L, respectively) dominated. DHEA was excreted mostly as sulfate (0-1 month of age: 184.5 nmol/L; 15-20 years of age: 1618.4 nmol/L) in all age groups. Cortisol was present predominantly as sulfate (mean: 173.8 nmol/L) in newborns. Levels of sulfated cortisol decreased with age, its glucuronidated form increased. The levels of free cortisol were relatively constant throughout childhood. Sex hormones were preferably excreted as glucuronides. In general, steroid hormone metabolites were conjugated to various extents with glucuronic acid or sulfuric acid and their ratio changed over lifetime.

Advancements in the gold standard: Measuring steroid sex hormones by mass spectrometry.

Sex hormones, such as testosterone and estrogens, play an essential role in regulating physiological and reproductive development throughout the lifetime of the individual. Although variation in levels of these hormones are observed throughout the distinct stages in life, significant deviations from reference ranges can result in detrimental effects to the individual. Alterations, by either an increase or decrease, in hormone levels are associated with physiological changes, decreased reproductive capabilities, and increased risk for diseases. Hormone therapies (HTs) and assisted reproductive technologies (ARTs) are commonly used to address these factors. In addition to these treatments, gender-affirming therapies, an iteration of HTs, are also a prominent treatment for transgender individuals. Considering that the effectiveness of these treatments relies on achieving therapeutic hormone levels, monitoring of hormones has served as a way of assessing therapeutic efficay. The need for reliable methods to achieve this task has led to great advancements in methods for evaluating hormone concentrations in biological matrices. Although immunoassays are the more widely used method, mass spectrometry (MS)-based methods have proven to be more sensitive, specific, and reliable. Advances in MS technology and its applications for therapeutic hormone monitoring have been significant, hence integration of these methods in the clinical setting is desired. Here, we provide a general overview of HT and ART, and the immunoassay and MS-based methods currently utilized for monitoring sex hormones. Additionally, we highlight recent advances in MS-based methods and discuss future applications and considerations for MS-based hormone assays.

A polyaromatic receptor with high androgen affinity.

Biological receptors distinguish and bind steroid sex hormones, e.g., androgen-, progestogen-, and estrogen-type hormones, with high selectivity. To date, artificial molecular receptors have been unable to discriminate between these classes of biosubstrates. Here, we report that an artificial polyaromatic receptor preferentially binds a single molecule of androgenic hormones, known as "male" hormones (indicated with m), over progestogens and estrogens, known as "female" hormones (indicated with f), in water. Competitive experiments established the binding selectivity of the synthetic receptor for various sex hormones to be testosterone (m) > androsterone (m) >> progesterone (f) > ß-estradiol (f) > pregnenolone (f) > estriol (f). These bindings are driven by the hydrophobic effect, and the observed selectivity arises from multiple CH-π contacts and hydrogen-bonding interactions in the semirigid polyaromatic cavity. Furthermore, micromolar fluorescence detection of androgen was demonstrated using the receptor containing a fluorescent dye in water.

Detection and analysis of 17 steroid hormones by ultra-high-performance liquid chromatography-electrospray ionization mass spectrometry (UHPLC-MS) in different sex and maturity stages of Antarctic krill (Euphausia superba Dana).

A sensitive and accurate method for determination of 17 endogenous and exogenous steroid hormones in Antarctic krill was developed. The method utilized UHPLC-MS in electrospray ionization mode (ESI). Samples were prepared by alkaline hydrolysis; sequential vortex extraction with ethyl acetate, methanol and acetonitrile; followed by a QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) clean-up method. The system suitability tests including theoretical plate number, resolution, repeatability, tailing factor proved the system's resolution and reproducibility that can meet the requirements of sample analysis. The developed method resulted in satisfactory recoveries that varied from 75.4%-110.6% and relative standard deviations (RSDs) that ranged from 3.1%-10.5%. The ranges of the limits of detection (LODs) and the limits of quantitation (LOQs) were 2-30 ng kg-1 and 10-100 ng kg-1, respectively. 14 hormones including cortisone, aldosterone, testosterone propionate, estriol, megestrol acetate, cortisone acetate, dexamethasone, testosterone, hydroxyprogesterone, nandrolone, prednisolone, cortisol, progesterone and estradiol were found in Antarctic krill. Other 3 hormones (Diethylstilbestrol, norethisterone and androsterone) were not detected. The levels of exogenous steroid hormones were much greater than those of endogenous steroid hormones, and the levels of exogenous glucocorticoids were much greater than those of exogenous sex hormones. The changes of hormones in different sex and maturity stages were also explored. Endogenous hormones might regulate the reproductive and development of Antarctic krill. The detected exogenous hormones suggests the potential for hormonal contamination in Antarctic waters that can affect organisms even affect human beings by food chain.

Watershed scale patterns in steroid hormones composition and content characters at a typical eutrophic lake in southeastern China.

Natural steroid hormones in the aquatic environment have attracted increasing attention because of their strong endocrine disrupting potency. Seven steroid hormones (estrone, 17α-estradiol, 17ß-estradiol, estriol, testosterone, androstenedione, and progesterone) were analyzed from surface water and sediment sampled from Chaohu Lake, its upstream rivers (the Hangbu River, Nanfei River, Shiwuli River, and Pai River), drainage from the adjacent farmland, and treated and untreated municipal sewage. Concentrations of the seven target steroid hormones ranged from below the detection limit (ND) to 69.5 ng L-1 in the water of Chaohu Lake and the upstream rivers. Three steroids-estrone, estriol, and 17α-estradiol-were found in relatively high residual concentrations in the water, with maximum concentrations of 69.5 ng L-1, 51.5 ng L-1, and 23.3 ng L-1, respectively. All of the target steroid hormones except estriol were detected in the sediment in concentrations of ND-16344 ng kg-1. The dominant steroid hormone in the sediment of Chaohu Lake and the upstream rivers was 17α-estradiol. In the Shiwuli River and the Pai River, the dominant steroid hormones (estrone and estriol) were the same as those in the untreated municipal sewage. This confirmed the deduction that untreated municipal sewage was the major source of steroid hormone residues in these two rivers. The main steroid hormone in the water of the Hangbu River and Chaohu Lake was 17α-estradiol, the same as that in the farmland drainage. In addition, 17α-estradiol was verified as the major factor in the contribution of farmland drainage to the pollution in these rivers. The water in the Nanfei River had high concentrations of estriol and 17α-estradiol. This indicates that both untreated municipal sewage and farmland drainage were the major sources. The discharge of steroid hormones from the four rivers to Chaohu Lake was approximately 75.1 kg year-1, with the highest contributor being 17α-estradiol (24 kg year-1). Therefore, based on the results of this study, the farmland drainage should be controlled to reduce the steroid hormone pollution in Chaohu Lake.

Human versus non-human sex steroid use in hormone replacement therapies part 1: Preclinical data.

Prior to 2002, hormone replacement therapy (HRT) was considered to be an important component of postmenopausal healthcare. This was based on a plethora of basic, epidemiological and clinical studies demonstrating the health benefits of supplementation with human sex steroids. However, adverse findings from the Women's Health Initiative (WHI) studies that examined the 2 major forms of HRT in use in the US at that time - Premarin (conjugated equine estrogens; CEE) and Prempro (CEE + medroxyprogesterone acetate; MPA), cast a shadow over the use of any form of HRT. Here we review the biochemical and physiological differences between the non-human WHI study hormones - CEE and MPA, and their respective human counterparts 17ß-estradiol (E2) and progesterone (P4). Preclinical data from the last 30 years demonstrate clear differences between human and non-human sex steroids on numerous molecular, physiological and functional parameters in brain, heart and reproductive tissue. In contrast to CEE supplementation, which is not always detrimental although certainly not as optimal as E2 supplementation, MPA is clearly not equivalent to P4, having detrimental effects on cognitive, cardiac and reproductive function. Moreover, unlike P4, MPA is clearly antagonistic of the positive effects of E2 and CEE on tissue function. These data indicate that minor chemical changes to human sex steroids result in physiologically distinct actions that are not optimal for tissue health and functioning.

Characterization of distinct ovarian isoform of crustacean female sex hormone in the kuruma prawn Marsupenaeus japonicus.

The eyestalk hormone, crustacean female sex hormone (CFSH), regulates the development of female secondary sexual characteristics in the blue crab Callinectes sapidus. After its discovery, several CFSH gene orthologs have been identified in some species of the suborder Pleocyemata as well. Similarly, in species of another suborder (Dendrobranchiata), an ortholog (Maj-CFSH) has been characterized as an eyestalk factor expressed in both females and males of the kuruma prawn, Marsupenaeus japonicus. In this study, another novel CFSH isoform was identified in the same species using cDNA cloning, expression analysis, and recombinant protein production. The isoform has "CFSH-family" structural characteristics but is dominantly expressed in the ovary, and was therefore designated as Maj-CFSH-ov. Its mRNA and protein levels in vitellogenic ovaries are higher than those in non-vitellogenic ovaries. In the vitellogenic ovary, both mRNA and protein expression of Maj-CFSH-ov are localized to oogonia and previtellogenic oocytes that occupy a small portion of vitellogenic ovaries, but not to the major developing oocytes. A vitellogenesis-inhibiting peptide of M. japonicus (Pej-SGP-I) reduced the expression of vitellogenin in incubated ovarian fragments, but not that of Maj-CFSH-ov. These results indicate that M. japonicus possesses two CFSH isoforms that are derived from distinct tissues, the central X-organ/sinus gland complex and peripheral ovaries. The expression profile of Maj-CFSH-ov suggests its involvement in some reproductive process other than vitellogenesis.

Attenuation of antimicrobial activity by the human steroid hormones.

Upon entering the human host, Staphylococcus aureus is exposed to endogenous steroid hormones. The interaction between S. aureus and dehydroepiandosterone (DHEA) results in an increased resistance to the host cationic defense peptide, ß-1 defensin, as well as vancomycin and other antibiotics that have a positive charge. The increased resistance to vancomycin is phenotypic and appears to correlate with a DHEA-mediated alteration in cell surface architecture. DHEA-mediated cell surface changes include alterations in: cell surface charge, surface hydrophobicity, capsule production, and carotenoid production. In addition, exposure to DHEA results in decreased resistance to lysis by Triton X-100 and lysozyme, indicating activation of murien hydrolase activity. We propose that DHEA is an interspecies quorum-like signal that triggers innate phenotypic host survival strategies in S. aureus that include increased carotenoid production and increased vancomycin resistance. Furthermore, this DHEA-mediated survival system may share the cholesterol-squalene pathway shown to be statin sensitive thus, providing a potential pathway for drug targeting.

[Aging and homeostasis. Sex hormones and aging.]

The decline in sex hormones along with aging is suggested to be involved in age-associated diseases such as cardiovascular diseases, osteoporosis, and dementia. The decrease of estrogen level after menopause is related to osteoporosis in addition to climacteric disturbance in women, while that of testosterone to sarcopenia, osteoporosis and late-onset hypogonadism in men. New treatment strategy is expected for targeting age-associated diseases against the decreased level of sex hormones.

Sex Hormones Function as Sex Attractant Pheromones in House Mice and Brown Rats.

Sex hormones of mammals control the expression of sexual characteristics and bodily functions. The male hormone testosterone and the female hormones progesterone and estradiol are known to occur in urine markings of mice. Here, we show that all three hormones are also present in urine of brown rats, and that they are effective sexual communication signals (pheromones) that elicit attraction behavior of prospective mates in both brown rats and house mice. When added as lures to trap boxes in field experiments, synthetic testosterone, for example, increased captures of adult female mice 15-fold, and a blend of progesterone and estradiol increased captures of male mice eightfold and male rats 13-fold. Remarkably, these hormones increased captures even though the food- and pheromone-based baits to which they were added had previously been shown to be superior to current commercial rodent attractants. We predict that these sex hormones will function as sex attractant pheromones in diverse taxa.

Androgens in women are essentially made from DHEA in each peripheral tissue according to intracrinology.

The objective is to review how the cell-specific amounts of intracellular androgens are all made in women from circulating dehydroepiandrosterone (DHEA) in each peripheral tissue, independently from the rest of the body. Following 500 million years of evolution, approximately three dozen cell-specific intracrine enzymes have been engineered in human peripheral tissues whereby the inactive sex steroid precursor DHEA mainly of adrenal origin is transformed into the appropriate minute intracellular amounts of androgens. These intracellular androgens are inactivated in the same cells, with no biologically significant release of active androgens in the circulation. The best estimate is that approximately 50% as much androgens are synthesized in women, compared to men of the same age. The problem with DHEA, however, the exclusive source of androgens in women of all ages, is that DHEA secretion has already decreased by an average of 60% at time of menopause and continues to decrease thereafter. The human-specific and highly sophisticated mechanisms of intracrinology permit each cell to control androgen availability according to its own needs independently from the remaining of the body. Such a mechanism is completely different from classical endocrinology well understood in men where testosterone of testicular origin is transported through the blood and has indiscriminate access to the androgen receptor (AR) in all AR-containing cells of the body. In men, both the endocrine and intracrine mechanisms are in operation while, in women, only the intracrine mechanisms responsible for intracellular formation from DHEA provide androgens.

Quantum-mechanical parameters for the risk assessment of multi-walled carbon-nanotubes: A study using adsorption of probe compounds and its application to biomolecules.

This work forwards new insights into the risk-assessment of multi-walled carbon-nanotubes (MWCNTs) while analysing the role of quantum-mechanical interactions between the electrons in the adsorption of probe compounds and biomolecules by MWCNTs. For this, the quantitative models are developed using quantum-chemical descriptors and their electron-correlation contribution. The major quantum-chemical factors contributing to the adsorption are found to be mean polarizability, electron-correlation energy, and electron-correlation contribution to the absolute electronegativity and LUMO energy. The proposed models, based on only three quantum-chemical factors, are found to be even more robust and predictive than the previously known five or four factors based linear free-energy and solvation-energy relationships. The proposed models are employed to predict the adsorption of biomolecules including steroid hormones and DNA bases. The steroid hormones are predicted to be strongly adsorbed by the MWCNTs, with the order: hydrocortisone > aldosterone > progesterone > ethinyl-oestradiol > testosterone > oestradiol, whereas the DNA bases are found to be relatively less adsorbed but follow the order as: guanine > adenine > thymine > cytosine > uracil. Besides these, the developed electron-correlation based models predict several insecticides, pesticides, herbicides, fungicides, plasticizers and antimicrobial agents in cosmetics, to be strongly adsorbed by the carbon-nanotubes. The present study proposes that the instantaneous inter-electronic interactions may be quite significant in various physico-chemical processes involving MWCNTs, and can be used as a reliable predictor for their risk assessment.

Magnetic porous carbon derived from a metal-organic framework as a magnetic solid-phase extraction adsorbent for the extraction of sex hormones from water and human urine.

An iron-embedded porous carbon material (MIL-53-C) was fabricated by the direct carbonization of MIL-53. The MIL-53-C possesses a high surface area and good magnetic behavior. The structure, morphology, magnetic property, and porosity of the MIL-53-C were studied by scanning electron microscopy, transmission electron microscopy, vibrating sample magnetometry, and N2 adsorption. With the use of MIL-53-C as the magnetic solid-phase extraction adsorbent, a simple and efficient method was developed for the magnetic solid-phase extraction of three hormones from water and human urine samples before high-performance liquid chromatography with UV detection. The developed method exhibits a good linear response in the range of 0.02-100 ng/mL for water and 0.5-100 ng/mL for human urine samples, respectively. The limit of detection (S/N = 3) for the analytes was 0.005-0.01 ng/mL for water sample and 0.1-0.3 ng/mL for human urine sample. The limit of quantification (S/N = 10) of the analytes were in the range of 0.015-0.030 and 0.3-0.9 ng/mL, respectively.

[Naming and classification of steroids and human stress ulcers. Articles of historic significance published by Hans Selye 70 years ago]. / A szteroidok elnevezése, felosztása és az emberi stresszfekélyek. Selye János 70 éve megjelent történelmi jelentoségu cikkei.

The name of Hans Selye is mostly known worldwide as the discoverer of stress reaction. Yet, he made numerous other seminal and clinically relevant discoveries. Namely, since he had a focused research on steroid hormones originating from the adrenal cortex that play a crucial role in stress response, he was the first who introduced about 70 years ago the first classification of steroids that is still valid nowadays. This is based on three objective facts: (a) the names of steroid groups are identical with their organ of origin (e.g., corticoids from the adrenal cortex, testoids/androgens from the testis); (b) chemical structures of the steroids are identical within a group (e.g., all corticoids have pregnane nucleus with 21 carbon atoms); and (c) the biological effects are homogenous within a group (e.g., all glucocorticoids exert catabolic effect, while androgens are anabolic). It should be emphasized that Selye also discovered in animal models the pro-inflammmatory effect of mineralocorticoids and the anti-inflammatory properties of glucocorticoids, about 8-10 years before Nobel Prize was awarded to a physician for the first clinical use of adrenocorticotrop hormone and cortisone. Last, but not least, Selye was the first who recognized about 70 years ago the occurence of stress ulcers in humans, based on clinical reports on the huge increase in the number of perforated gastric anti-duodenal ulcers during bombings of London in World War II. The subsequent ulcer research by Selye`s former students and their contemporaries resulted in the recognition of anti-duodenal ulcer effect of dopamine, and the central gastroprotective actions of thyreotrop releasing hormone and endogenous opioids. Thus, Hans Selye made much more contributions to medical science and clinical practice than 'just' the discoverer of biologic stress response.

Determination of steroid sex hormones in real matrices by bar adsorptive microextraction (BAµE).

In the present work, the development of a novel analytical approach which combines a miniaturized bar adsorptive microextraction device with a micro-liquid desorption in one single step, followed by high performance liquid chromatography with diode array detection (BAµE-µLD/HPLC-DAD), is proposed for the determination of trace levels of nine steroid hormones (estriol, 17ß-estradiol, 17α-estradiol, 19-northisterone, 17α-ethynylestradiol, estrone, D-(-)-norgestrel, progesterone and mestranol) in environmental and biological matrices. From the comparison of ten different coating phases (five polymeric and five activated carbon sorbents), the modified pyrrolidone polymer (P2) showed the best compromise between selectivity and efficiency. Assays performed through BAµE(P2, 1.3mg)-µLD(100µL)/HPLC-DAD on 25mL of ultrapure water samples spiked at the 6.0µg/L level, yielded recoveries ranging from 93±9% to 101±8%, under optimized experimental conditions. The analytical performance showed convenient detection (50.0-100.0ng/L) and quantification limits (165.0-330.0ng/L), as well as good linear dynamic ranges (0.2-24.0µg/L) with remarkable determination coefficients (r(2)>0.9968). Excellent repeatability were also achieved through intraday (RSD<14%) and interday (RSD<12%) experiments. The application of the proposed analytical approach on environmental water and urine samples, using the standard addition methodology (SAM), revealed good linearity and sensitivity at trace level, with the detection of some of the target compounds. In short, the miniaturization of the analytical device for microextraction combined with the minimization of the solvent volume for back-extraction in one single step demonstrated remarkable performance, increasing the enrichment factor, being simultaneously more easier to implement and environment friendly.

[Toxic effects of mixture of volatile organic compounds on mice testis related enzymes and hormones].

OBJECTIVE: To evaluate the toxic effects of mixture of volatile organic compounds (VOCs) on Mice Testis related enzymes and hormones. METHODS: After determining the median lethal dose (LD50) of VOCs using the acute toxicity test, 40 male clean inbred Kunming mice were assigned to 1/8 LD50 VOCs exposure group, 1/4 LD50 VOCs exposure group, and 1/2 LD50 VOCs exposure group, as well as positive control group with cyclophosphamide (60 mg/kg) and negative control group with tea oil, with 8 mice in each group. The mice were intraperitoneally injected with respective agents for 5 days. The levels of testis testosterone, estradiol, follicle stimulating hormone, and luteinizing hormone were determined by ELISA. Meanwhile, the activity of testicular marked enzymes such as lactate dehydrogenase, gamma-glutamyl transpeptidase, acid phosphatase, and glucose-6-phosphate dehydrogenase were examined. RESULTS: Compared with the negative control group, the 1/8 LD50 exposure group had a significantly increased testis coefficient (P<0.05). Both the activity of testicular marked enzymes and the levels of testicular sex hormones in all exposure groups showed significant downward trends with increasing VOC doses compared with those in the negative control group (P<0.05). CONCLUSION: VOCs have obvious toxicity to mouse testis by changing the levels of testicular sex hormones and the activity of testicular marked enzymes.

Proteomic-coupled-network analysis of T877A-androgen receptor interactomes can predict clinical prostate cancer outcomes between White (non-Hispanic) and African-American groups.

The androgen receptor (AR) remains an important contributor to the neoplastic evolution of prostate cancer (CaP). CaP progression is linked to several somatic AR mutational changes that endow upon the AR dramatic gain-of-function properties. One of the most common somatic mutations identified is Thr877-to-Ala (T877A), located in the ligand-binding domain, that results in a receptor capable of promiscuous binding and activation by a variety of steroid hormones and ligands including estrogens, progestins, glucocorticoids, and several anti-androgens. In an attempt to further define somatic mutated AR gain-of-function properties, as a consequence of its promiscuous ligand binding, we undertook a proteomic/network analysis approach to characterize the protein interactome of the mutant T877A-AR in LNCaP cells under eight different ligand-specific treatments (dihydrotestosterone, mibolerone, R1881, testosterone, estradiol, progesterone, dexamethasone, and cyproterone acetate). In extending the analysis of our multi-ligand complexes of the mutant T877A-AR we observed significant enrichment of specific complexes between normal and primary prostatic tumors, which were furthermore correlated with known clinical outcomes. Further analysis of certain mutant T877A-AR complexes showed specific population preferences distinguishing primary prostatic disease between white (non-Hispanic) vs. African-American males. Moreover, these cancer-related AR-protein complexes demonstrated predictive survival outcomes specific to CaP, and not for breast, lung, lymphoma or medulloblastoma cancers. Our study, by coupling data generated by our proteomics to network analysis of clinical samples, has helped to define real and novel biological pathways in complicated gain-of-function AR complex systems.