Endocrine Compounds Research Articles

6951 European survey of diagnosis and management of the polycystic ovary syndrome: full report on the ESE PCOS Special Interest Group’s 2023 Questionnaire

Abstract Disclosure: S. Livadas: None. B.O. Yildiz: None. G. Mastorakos: None. A. Gambineri: None. D.L. Pignatelli: None. F. Giorgino: None. M. Andersen: None. B.M. Obermayer-Pietsch: None. D.P. Macut: None. Background: Although polycystic ovary syndrome (PCOS) constitutes a common endocrinopathy, there are several issues which confuse clinicians during everyday practice.Objective: To define the current status of knowledge of the full spectrum of PCOS among European endocrinologists. Methods: A detailed questionnaire comprising 41 items covering various aspects of diagnosis and management of women with PCOS was shared via web among members of European Society of Endocrinology. Results: 505 European endocrinologists (64% females), with a mean age of 47±11.6 years answered the questionnaire. Rotterdam criteria were used by 85% of endocrinologists. The most frequent age at referral was between 20-40 years (87.1%). A quarter of doctors have access to mass spectrometry for the evaluation of androgen levels. Extended metabolic profile was part of workup by the vast majority, but there was significant uncertainty regarding the diagnosis of chronic anovulation. Diabetes, gestational or type 2, was considered a major consequence for women with PCOS and screening is carried out regardless of BMI status. Lifestyle modification and metformin are considered a standard approach of all participants and oral contraceptives are a standard therapeutic modality, but there is significant discrepancy on the duration of treatment.Conclusions: Rotterdam diagnostic criteria are currently an established diagnostic approach within the European endocrine community. This updated survey showed an advance in using steroid profiling for diagnosis and strong position on recognizing PCOS as a metabolic condition with potential broader consequences. Therapeutic aspects are currently changed into the need for more lifestyle intervention involvement and the use of metabolic therapies either as monotherapy or in combination with standard hormonal compounds. Presentation: 6/3/2024

6951 European survey of diagnosis and management of the polycystic ovary syndrome: full report on the ESE PCOS Special Interest Group’s 2023 Questionnaire

Abstract Disclosure: S. Livadas: None. B.O. Yildiz: None. G. Mastorakos: None. A. Gambineri: None. D.L. Pignatelli: None. F. Giorgino: None. M. Andersen: None. B.M. Obermayer-Pietsch: None. D.P. Macut: None. Background: Although polycystic ovary syndrome (PCOS) constitutes a common endocrinopathy, there are several issues which confuse clinicians during everyday practice.Objective: To define the current status of knowledge of the full spectrum of PCOS among European endocrinologists. Methods: A detailed questionnaire comprising 41 items covering various aspects of diagnosis and management of women with PCOS was shared via web among members of European Society of Endocrinology. Results: 505 European endocrinologists (64% females), with a mean age of 47±11.6 years answered the questionnaire. Rotterdam criteria were used by 85% of endocrinologists. The most frequent age at referral was between 20-40 years (87.1%). A quarter of doctors have access to mass spectrometry for the evaluation of androgen levels. Extended metabolic profile was part of workup by the vast majority, but there was significant uncertainty regarding the diagnosis of chronic anovulation. Diabetes, gestational or type 2, was considered a major consequence for women with PCOS and screening is carried out regardless of BMI status. Lifestyle modification and metformin are considered a standard approach of all participants and oral contraceptives are a standard therapeutic modality, but there is significant discrepancy on the duration of treatment.Conclusions: Rotterdam diagnostic criteria are currently an established diagnostic approach within the European endocrine community. This updated survey showed an advance in using steroid profiling for diagnosis and strong position on recognizing PCOS as a metabolic condition with potential broader consequences. Therapeutic aspects are currently changed into the need for more lifestyle intervention involvement and the use of metabolic therapies either as monotherapy or in combination with standard hormonal compounds. Presentation: 6/3/2024

Gibberellin 2-oxidase 1(CsGA2ox1) involved gibberellin biosynthesis regulates sprouting time in camellia sinensis

BackgroundTea is an important cash crop and buds are its main product. To elucidate the molecular mechanism of the sprouting time of tea plants, ‘Yuchunzao’, which was an early sprouting tea cultivar, was studied. ‘Echa 1’, sprout one week later than ‘Yuchunzao’ in spring, was used as the control.ResultsA total of 26 hormonal compounds and its derivatives in tea plants were qualified by using Ultra Performance Liquid Chromatography-Tandem mass spectrometry (UPLC-MS/MS). The result showed that GA20, GA3 and ICA were significantly different in ‘Yuchunzao’ than in ‘Echa 1’, with GA20 and GA3 up-regulated and ICA down-regulated. Based on the Illumina platform, transcriptome analysis revealed a total of 5,395 differentially expressed genes (DEGs). A diterpenoid biosynthesis related gene, gibberellin 2-oxidase 1 (CsGA2ox1), was downregulated in ‘Yuchunzao’ compared to ‘Echa 1’. CsGA2ox1 regulate the transformation of GA different forms in plants. The relative expression of CsGA2ox1 showed an adverse trend with the content of GA20 and GA3. Our results suggest that down regulation of CsGA2ox1 resulted in the accumulation of GA3 and GA20, and then promoted sprout of ‘Yuchunzao’.ConclusionThis study provides theoretical basis of tea plants sprout and guides the tea breeding in practice.

Simultaneous determination of 61 hormones in water by solid phase extraction-ultra performance liquid chromatography-tandem mass spectrometry

Concerns over the emergence of steroid hormones as pollutants in water have grown. Steroid hormone compounds present challenges in the simultaneous detection of total residual hormones owing to their analogous structures and diverse types. In this study, we established a rapid and high-throughput continuous online method based on solid phase extraction (SPE) coupled with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for the simultaneous determination of 61 hormone components, including 48 glucocorticoids, 1 mineralocorticoid, 4 androgens, and 8 progesterones, in water. Various SPE columns were investigated to assess their extraction efficiency for enriching and purifying target compounds in a large sample volume (1 L). An HC-C18 SPE column was selected because of its superior performance. Acetonitrile was used as a washing solution during SPE to ensure that the majority of the tested substances achieved recoveries exceeding 70% and effectively avoid interferences from water-soluble components. Various C8 and C18 columns were tested, and the optimal HPLC conditions for hormone retention were established. We systematically evaluated different UPLC columns and mobile phases, including methanol-water and acetonitrile-water systems with 0.1% formic acid added to the aqueous phase. The optimized UPLC conditions were as follows: BEH C18 column (100 mm×2.1 mm, 1.7 μm); column temperature, 40 ℃; flow rate, 0.3 mL/min; injection volume, 5 μL; mobile phase A: 0.1% formic acid aqueous phase; mobile phase B: acetonitrile. Gradient elution was performed as follows: 0-0.5 min, 30%B; 0.5-15.0 min, 30%B-75%B; 15.0-18.0 min, 75%B-98%B; 18.0-19.0 min, 98%B; 19.0-19.1 min, 98%B-30%B; 19.1-20.0 min, 30%B. Both positive- and negative-ion modes were explored in the UPLC-MS/MS experiment to obtain the full scan of the parent ions, and positive mode was finally selected for electrospray ionization (ESI). Two product ions exhibiting strong signals and minimal interference were selected for quantitative and qualitative ion analyses, using an external standard method for quantification. MS/MS was performed in positive-ion (ESI+) mode with multiple reaction monitoring (MRM) scanning. The MS/MS parameters were as follows: atomizing gas pressure, 379 kPa; curtain air pressure, 241 kPa; spray voltage, 5500 V; desolvation temperature, 550 ℃; collision exit voltage (CXP), 13 V; intake voltage (EP), 10 V; and residence time of each ion pair, 0.5 ms. Other instrument settings, such as the collision energy and declustering voltage, were also optimized. The 61 hormones exhibited excellent linear relationships within their corresponding concentration ranges, with correlation coefficients greater than 0.99. The method detection limits (MDLs) were in the range of 0.05-1.50 ng/L. The average recoveries of the 61 hormones across three spiked levels ranged from 62.3% to 125.2%, with relative standard deviations (RSDs, n=6) of 1.1%-10.5%. Using this method, we successfully detected 10 hormone components (cortisone, fluticasone propionate, ciclesonide, betamethasone dipropionate, clobetasone butyrate, diflucortolone valerate, halobetasol propionate, isoflupredone, difluprednate, and hydroxyprogesterone caproate) in various surface water and groundwater samples collected from the Taihu Basin region. The SPE-UPLC-MS/MS method presented in this paper is simple, highly sensitivity, and exceptionally accurate. Thus, it exhibits promising potential for tracing targeted hormone residues in water and will be of great value in monitoring and ensuring water safety. Finally, a regional analysis was conducted on the hormone levels in water, and suggestions were made for the targeted treatment of hormone residues in future sewage treatment processes.

European survey of diagnosis and management of the polycystic ovary syndrome: full report on the ESE PCOS Special Interest Group's 2023 Questionnaire.

Although polycystic ovary syndrome (PCOS) is a very common endocrinopathy, there are several issues related to this disorder which perplex clinicians in their everyday practice. To determine the current state of knowledge among European endocrinologists concerning the full spectrum of PCOS. An online survey comprising 41 items covering various aspects of PCOS diagnosis and management was distributed to members of the European Society of Endocrinology. A total of 505 European endocrinologists (64% females), with a mean age of 47 ± 11.6 years, participated in the survey. The Rotterdam criteria were the primary diagnostic tool for 85% of respondents. Most referrals (87.1%) occurred between ages 20 and 40 years. Twenty-five percent of physicians have access to mass spectrometry for the evaluation of androgen levels. While an extended metabolic profile was commonly employed as part of the workup, there was uncertainty regarding chronic anovulation diagnosis. Diabetes, including gestational or type 2, was recognized as a significant risk factor with universal screening irrespective of BMI status. Lifestyle modification and metformin were considered as standard interventions by all participants alongside oral contraceptives, though there was significant discrepancy in treatment duration. The Rotterdam diagnostic criteria are widely adopted for PCOS diagnosis among European endocrinologists. The current updated survey shows an emphasis on steroid profiling as an important part of diagnostic workup and a strong position held for recognition of PCOS as a metabolic condition with potentially serious implications. Current therapy thus shifted to the demand for prioritizing lifestyle interventions and metabolic therapies, either as monotherapy or in combination with standard hormone compounds.

Elucidating the effect of levothyroxine and triiodothyronine on methylglyoxal derived stress.

Methylglyoxal (MG) is the most potent precursor during the formation of the advanced glycation end products (AGEs). MG-dependent glycative stress contributes to pathogenesis of diabetes, age-related disorders, and cancer. There is a great need to study the reduction process of glycative stress for effective management of metabolic disorders. From natural compounds to synthetic drugs, each element contributes to the reduction of glycative stress. Previously, it was established that the lowering of uric acid, low-density lipoprotein cholesterol, and urine albumin excretion rate, as well as reducing total oxidative stress, were all achieved more effectively with a levothyroxine regimen. Still, there is no such study found that supports the MG-dependent glycative stress reduction with thyroid hormone compound. Our study aims to investigate the effects of T3 and T4 on MG-dependent glycative stress. The antiglycation effect was assayed through NBT assay, DNPH assay, ELISA, and fluorescence spectrophotometer. The intracellular reduction in reactive oxygen species (ROS) has been estimated through confocal microscopy. The results revealed an effective reduction in the formation of AGEs adducts and intracellular ROS formation. The investigation concludes AGEs formation was suppressed using these compounds, although in vivo and rigorous clinical trials are required in order to verify these findings.

Sugammadex and oral contraceptives.

This review article explores the evidence regarding sugammadex (MSD Australia) and its potential interaction with hormonal contraceptives. The impact of recent clinical trials and review articles is examined. Recent clinical data suggest that the interaction between sugammadex and estrogen and progesterone concentrations may not be clinically significant and may confer some protection against ovulation. There are no clinical trials reporting interactions between sugammadex and the exogenous hormonal compounds found in oral contraceptive pills. The method of contraception is an important consideration, as sugammadex theoretically affects oral and nonoral, and combined versus single agent methods differently. Two large retrospective database studies have reported two cases of pregnancy postoperatively in patients on hormonal contraceptives whose anesthetic included sugammadex. Strong clinical evidence to support or refute claims of a significant impact of sugammadex on contraceptive efficacy in women on contraception is lacking. The existing evidence does not suggest a basis for concern regarding the impact of sugammadex on contraception in the perioperative setting.

Sex chromosome complement interacts with gonadal hormones in determining regional-specific neuroactive steroid levels in plasma, hippocampus, and hypothalamus. A study using the four core genotype mouse model

An important aspect of the neuromodulatory and neuroprotective actions exerted by neuroactive steroids is that they are sex-specific, as determined by the sexually dimorphic levels of these molecules in plasma and the nervous tissue. Thus, the identification of the factors that generate the sex-dimorphic levels of neuroactive steroids may be crucial from a neuroprotectant perspective. The main driver for sex determination in mammals is the SRY gene and the subsequent presence of a specific gonad: testes for males and ovaries for females, thus producing hormonal compounds, primarily androgens and estrogens, respectively. Nowadays, it is well established that despite the relevance of gonads, other factors control sexual features, and, among them, sex chromosome complement is highly relevant. In this study, neuroactive steroids were evaluated by liquid chromatography-tandem mass spectrometry in the hypothalamus, the hippocampus, and plasma of the four core genotype mouse model, to determine the relative contribution of sex chromosome complement and gonads in determining their sex dimorphic levels. The data obtained reveal that although gonads are the main contributing factor for sex differences in neuroactive steroid levels, the levels of some neuroactive steroids, including testosterone, are also influenced in brain and plasma by tissue-specific actions of sex chromosomes. The data presented here adds a new piece to the puzzle of steroid level regulation, which may be useful in designing sex-specific neuroprotective approaches to pathological conditions affecting the nervous system.

(191) Contraception: How Ecology and Gender Equality are the New Deal

Abstract Introduction Since more than 10 years health care providers often hear from their patients that they want a natural contraception without hormones because of their potentials side-effects on bodies and environment by water contamination. In fact numerous scientific articles available on the web etablish that steroid hormones in wastewater are micropollutants and endocrine disruptors, especially wastewater treatment plants can not eradicate synthetic progestins after excretion. Nevertheless the situation remains paradoxical since the morning-after pill contains up to 50 times more hormones than the daily pill and is widely used, in France 2,2 millions of morning-after pills are sold each year. Looking for "green solutions", users are turning to menstruations apps, but are they aware of the pollution generated by materials and operations of digital industries? This is another paradox in the new contraceptive behaviours. Objective know different possible causes of contraceptive refusal take into account potential environmental effects when prescribing contraceptives and opt for the least polluting solutions. be aware of the environmental impact of digital health techniques take account of claims for gender-equal contraception Methods Literature review on PubMed for the 12 last months, Google Scholar since 2 years and mainstream media with the keywords: hormonal contraception and water pollution contraception and ecology Contraceptives residues and health Data Centers pollution contraception and gender equality Male contraception Results On Google Scholar numerous articles deal with the pollution of waste water and drinking water by steroid hormones at concentrations ranging from ng/L to g/L. Synthetic hormone compounds toxicity is widely recognized and can affect cell metabolism, causing various diseases such as obesity, infertility, endocrine disorders, even neoplasia or neuro-degenerative disease, although the effect of environmental synthetic hormones on the nervous system is still poorly documented. These data are public and can influence contraceptive behaviours On PubMed the focus is on endocrine-disrupting chemicals such as phtalates, dioxins, polychlorinated-biphenyl, biphenol A and analogs, I found only 2 surveys about residues of synthetic human hormones. 21 relevant surveys are found on PubMed about gender equality in contraception: these studies show an emerging interesr among men in controlling their fertilizing power, but a low support of pharmaceutical industries, leading them to use restrictive methods such as thermal contraception. Conclusions This work aims to highlight the contraceptive shift in progress. Consumers claim an ecological position, but use methods that contradict their stated convictions, such as morning-after pill or menstruations app causing digital pollution. Men want to involve themself in their reproductive health but are they heard enough? A low-risk, inexpensive and non-pollutante method such as vasectomy is still subject to false beliefs due to a lack of information and education, male hormonal contraception remains anecdotal (only in France). These observations highlight the role of the health care providers in sexual and reproductive health education and counselling, and of the industry's support, listening to people. Disclosure No.

Biochemical and allelopathic features of Adonis vernalis, Allium ursinum, and Leucojum vernum in the M.M. Gryshko National Botanical Garden of the NAS of Ukraine

The article presents the results of a study on the content and dynamics of the accumulation of biogenic elements and brassinolides in plants of Adonis vernalis, Allium ursinum, and Leucojum vernum in Kyiv, Ukraine. Data is provided on allelopathic activity, content of macro- and microelements, phenolic compounds, and laccase activity in plants and the rhizosphere soil under the conditions of the M.M. Gryshko National Botanical Garden of the National Academy of Sciences of Ukraine (NBG). The plants from the collection of the NBG were used as objects of study in field experiments. The content of biogenic elements in plant tissues and soil was analyzed using an inductively coupled plasma spectrometer. The allelopathic analysis of soil was conducted using a direct bioassay method with Lepidium sativum seedlings as the test object. Phenolic compounds were extracted from the soil using the ion exchange (desorption) method. The content of brassinosteroids was measured spectrophotometrically at a wavelength of 450 nm. The content of laccase was measured spectrophotometrically at a wavelength of 530 nm.The results demonstrate that model plant species employ a wide range of physiological mechanisms throughout the vegetation period to enhance their resistance to abiotic factors. These mechanisms include maintaining potassium and calcium balance and utilizing hormonal compounds. Plants have been proven to have compensatory mechanisms in response to stress factors, substituting one biochemical marker of resistance with another. Both, brassinosteroids and silicon, contribute to the adaptive capacity of organisms.

Attenuated total reflection Fourier-transform infrared spectroscopy for the prediction of hormone concentrations in plants.

Plant hormones are important in the control of physiological and developmental processes including seed germination, senescence, flowering, stomatal aperture, and ultimately the overall growth and yield of plants. Many currently available methods to quantify such growth regulators quickly and accurately require extensive sample purification using complex analytic techniques. Herein we used ultra-performance liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS) to create and validate the prediction of hormone concentrations made using attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectral profiles of both freeze-dried ground leaf tissue and extracted xylem sap of Japanese knotweed (Reynoutria japonica) plants grown under different environmental conditions. In addition to these predictions made with partial least squares regression, further analysis of spectral data was performed using chemometric techniques, including principal component analysis, linear discriminant analysis, and support vector machines (SVM). Plants grown in different environments had sufficiently different biochemical profiles, including plant hormonal compounds, to allow successful differentiation by ATR-FTIR spectroscopy coupled with SVM. ATR-FTIR spectral biomarkers highlighted a range of biomolecules responsible for the differing spectral signatures between growth environments, such as triacylglycerol, proteins and amino acids, tannins, pectin, polysaccharides such as starch and cellulose, DNA and RNA. Using partial least squares regression, we show the potential for accurate prediction of plant hormone concentrations from ATR-FTIR spectral profiles, calibrated with hormonal data quantified by UHPLC-HRMS. The application of ATR-FTIR spectroscopy and chemometrics offers accurate prediction of hormone concentrations in plant samples, with advantages over existing approaches.

Construction and analysis of the tapping panel dryness-related lncRNA/circRNA-miRNA-mRNA ceRNA network in latex of Hevea brasiliensis

Tapping panel dryness (TPD) results in a severe reduction in latex yield in Hevea brasiliensis. However, the molecular regulatory mechanisms of TPD occurrence are still largely unclear. In this study, whole-transcriptome sequencing was carried out on latex from TPD and healthy trees. In total, 7078 long noncoding RNAs (lncRNAs), 3077 circular RNAs (circRNAs), 4956 miRNAs, and 25041 mRNAs were identified in latex, among which 435 lncRNAs, 68 circRNAs, 320 miRNAs, and 1574 mRNAs were differentially expressed in the latex of TPD trees. GO and KEGG analyses indicated that plant hormone signal transduction, MAPK signaling pathway, and ubiquitin-mediated proteolysis were the key pathways associated with TPD onset. Phytohormone profiling revealed significant changes in the contents of 28 hormonal compounds, among which ACC, ABA, IAA, GA, and JA contents were increased, while SA content was reduced in TPD latex, suggesting that hormone homeostasis is disrupted in TPD trees. Furthermore, we constructed a TPD-related competitive endogenous RNA (ceRNA) regulatory network of lncRNA/circRNA-miRNA-mRNA with 561 edges and 434 nodes (188 lncRNAs, 5 circRNAs, 191 miRNAs, and 50 mRNAs) and identified two hub lncRNAs (MSTRG.11908.1 and MSTRG.8791.1) and four hub miRNAs (hbr-miR156, miR156-x, miRf10477-y, and novel-m0452-3p). Notably, the lncRNA-miR156/157-SPL module containing three hubs probably plays a crucial role in TPD onset. The expression of network hubs and the lncRNA-miR156/157-SPL module were further validated by qRT-PCR. Our results reveal the TPD-associated ceRNA regulatory network of lncRNA/circRNA-miRNA-mRNA in latex and lay a foundation for further investigation of molecular regulatory mechanisms for TPD onset in H. brasiliensis.

Extraction and determination methods of phthalates

Phthalic acid esters (PAEs) are environmental hormone compounds, commonly known as plasticizers. After long-term accumulation, it will lead to deformities, cancer, mutations and other diseases in the human body, which will seriously endanger human health. Due to the increasing attention paid to phthalate pollutants in society, high efficiency, accuracy and no secondary pollution are difficult points in extraction and determination methods. This paper summarizes the research progress of extraction and determination methods of phthalates at home and abroad from 2010 to 2022, mainly summarizes the extraction methods of phthalates in various environments and the methods of chromatography, Measurement methods such as mass spectrometry and spectroscopy. Compared the advantages and disadvantages of several extraction and determination methods, and proposed the prospect of improving extraction and detection methods.

The Configuration of Some Hormonal Compounds Play an Important Role in Pharmacological Action (Agonist, Antagonist, Active, More Active)

In this research we explain the configuration of some hormonal compounds and its effect of these configurations on the activity for example: Dexamethasone and Betamethasone, where the difference on the carbon number 16, where the methyl group take alpha position and beta position respectively where the beta is more active than dexa, estradiol and alfatradiol have Hydroxyl group at carbon number 17 in beta position and alpha position respectively, where the estradiol is natural estrogen while alfatradiol is in contrary to estradiol has no estrogenic effect, clomiphene have 2 configurations Z (cis) and E (trans), zuclophene and eneclophene, the enclophene isomer has antiestrogenic and the zuclophene has estrogenic activity, tamoxifen also has 2 configuration cis and trans which have estrogenic and antiestrogenic respectively, Alpha-Medroxyprogestrone is more active than Beta-Medroxyprogestrone in birth control, fluticasone and budesonide both are used in R and S epimers.

National medical eligibility criteria for contraceptive use 2023: focus on women's reproductive health: A review

Reproductive health care involves helping families in this area with the provision of the necessary information. This is recognized as a key step in improving the health of men, women and children and is a fundamental human right. Everyone has the right to have access to a correct understanding of reproductive health care, choice of contraceptive method and the opportunity to benefit from scientific progress in this area. Over the past 35 years, significant progress has been made in the development of new contraceptives: the content of hormones in combined oral contraceptives has significantly decreased, inert intrauterine contraceptives have been replaced by more progressive, levonorgestrel-containing ones including low-dose generation. Combined hormonal compounds were synthesized in the form of injections, patches and vaginal rings, progestogen implantable and injectable contraceptives. Women who use contraceptives must take into account the state of health (including the presence of endocrine pathology) in order to use them without harm to their health. Based on the World Health Organization Medical Eligibility Criteria for Contraceptive Methods 2015, the Russian Medical Eligibility Criteria for Contraceptive Methods 2023 is designed to become an important tool in daily practice of doctors.

The role of local expression of hormone receptors in the genesis of idiopathic tracheal stenosis.

Tracheal stenosis in adults is usually the result of mechanical injuries either from direct trauma, tracheotomy or intubation. Idiopathic stenosis in the cricotracheal section is a rare condition and occurs almost exclusively in females. Therefore, an influence of the female sexual hormones estrogen and progesterone has been assumed previously. Tracheal specimens of 27 patients who received tracheal resection for either idiopathic tracheal stenosis (ITS) (n=11) or posttraumatic tracheal stenosis (PTTS) (n=16) between 2008 and 2019 in our surgical department were included and retrospectively analyzed. Immunohistochemical staining of tracheal specimens concerning the hormone receptor status of progesterone and estrogen was performed. While post-tracheotomy stenosis occurred in males (n=6) as well as in females (n=10), none of the patients with idiopathic stenosis were males. All of the idiopathic stenosis (n=11; 100%) showed a strong expression of the estrogen receptors (ERs) in the fibroblasts and expression of progesterone receptors (PRs) in fibroblasts in 8 of 11 (72.7%). In the post-tracheotomy patients, only 3/16 (18.8%) showed slight staining of PRs and 6/16 (37.5%) of ERs. Of those, only one male patient presented with expression of ERs and PRs and another male patient presented with isolated PRs. Oral intake of hormone compounds was seen in 11/27 (40.7%) patients: 7/11 (63.6%) in the ITS group and 4/16 (25%) in the PTTS (noteworthy that the PTTS group included 6 male patients). Although the number of patients is small, our results show that the expression of female sexual hormone receptors in the fibroblasts of the trachea is a persistent finding in ITS. Surgery provided good results with a favorable long-term outcome without recurrence of stenosis for ITS and PTTS. Further investigation with a special focus on hormones is needed to assist in the prevention of this rare disease.

A novel perspective to investigate how nanoselenium and melatonin lengthen the cut carnation vase shelf

Nano-selenium (nano-Se) and melatonin (MT) applications confirmed to boost plant growth and resistance. The mechanism of various ratios of nano-Se and MT foliar application postpone the senescence of fresh cut carnation flowers and improve vase life remains unclear. In this study, a combined effect with nano-Se (nano-Se5, 5 mg/L) and MT(MT1, 1 mg/L) was preferable to the control, nano-Se, and MT treatment alone when it came to delaying flower senescence. They enhance the antioxidant ability of carnation flowers by lowering MDA and H2O2 levels, raising SOD and POD concentrations, and lowering procyanidins biosynthesis (catechins and epicatechin). Inducing the biosynthesis of hormonal compounds (salicylic acid, jasmonic acid, and abscisic acid), their combination also boosted the growth of carnations. Biofortification with nano-Se and MT substantially increased the amounts of key lignin biosynthesis pathway metabolites (L-phenylalanine, p-hydroxycinnamic acid, p-coumaric acid, perillyl alcohol, p-Coumaryl alcohol, and cinnamic acid), which may increase stem cellular thickness and facilitate water absorption and transmission. The study hypothesizes that nano-Se and MT synergistic applications act as a new efficient non-toxic preservative to extend the vase life and improve the decorative value of carnations.

ENZYME ACTIVITY OF CARBOHYDRATE AND AMINOACID METABOLISM, LIPID PEROXIDATION IN CARP TISSUES UNDER THE EFFECT OF 19-NORTESTOSTERONE

The aim of the study was to investigate the activity of a number of enzymes of carbohydrate metabolism, individual amino acids, creatine phosphate, inorganic phosphorus and lipid peroxidation (LPO) in the hepatopancreas and blood plasma of carp (Cyprinus carpio L.) under the influence of the synthetic steroid 19-norethisterone. The experiments were conducted on two-year-old carps, which were kept for 24 hours in aquariums with different concentrations of 19-nortestosterone in water at optimal temperature, oxygen content and pH. The activity of lactate dehydrogenase (LDH), alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatine phosphokinase (CPK) in the supernatant fraction of hepatopancreatic homogenate was monitored by UV-kinetic and colourimetric methods. The activity of isocitrate dehydrogenase (ICDH) was determined spectrophotometrically by the accumulation of NADPH, and lactate dehydrogenase (LDH), glucose-6-phosphate dehydrogenase (Gl-6-PDH) and glucose-6-phosphatase (Gl-6-Phase) - by the accumulation of NADH in the reactions of conversion of the corresponding substrates. Conventional biochemical methods were used to determine the concentration of diene conjugates, malondialdehyde (MDA), lipid hydroperoxides, catalase (CAT) and superoxide dismutase (SOD) activity in blood plasma and supernatant fraction of hepatopancreatic homogenate. Conventional biochemical methods were used to determine the concentration of diene conjugates, malondialdehyde (MDA), lipid hydroperoxides, catalase (CAT) and superoxide dismutase (SOD) activity in blood plasma and the supernatant fraction of hepatopancreatic homogenate. It was found that fish exposed to water with a concentration of synthetic steroid 19-nortestosterone of 50 µg/dm3 for 24 hours had higher levels of MDA and diene conjugates, increased activity of CAT and SOD in blood plasma, increased activity of ALP, ALT, AST, CK, CAT and SOD in the hepatopancreatic supernatant fraction, and LDH remained unchanged, as well as the level of lipid hydroperoxides in blood plasma. Under the influence of 19-nortestosterone, the activity of Gl-6-FDG and IDDG in the mitochondrial fraction of carp hepatopancreas increases, and that of Gl-6-phase in the supernatant. At a concentration of 19-nortestosterone in water of 200 μg/dm3, the content of lipid hydroperoxides in the blood plasma of carp increases, and in the hepatopancreatic supernatant fraction - diene conjugates and SOD activity, while ALP and CPK decrease relative to their values in fish with a low level of xenobiotic in water. The content of MDA, diene conjugates, as well as the activity of CAT, SOD, ALP, CPK and LDH in blood plasma, IDH and Gl-6-PDH in the mitochondrial fraction and Gl-6-Phase and LDH in the hepatopancreatic supernatant did not change, but remained higher than in the control. It was concluded that 19-nortestosterone stimulates carbohydrate metabolism, amino acids alanine and aspartate transamination, creatine phosphate conversion, antioxidant enzyme activity and lipid peroxidation in the hepatopancreas of two-year-old carp. The obtained results deepen the understanding of the mechanism of action of synthetic steroids in bony fish, and the differences in enzyme activity and lipid peroxidation can be important criteria for ecotoxicological assessment of water bodies contaminated with hormonal compounds.

Expression of estrogen receptors, PELP1, and SRC in human spermatozoa and their associations with semen quality

Sperm cells are target cells for both estrogens and xenoestrogens. Due to the specific structure of spermatozoa, these hormonal compounds may act on sperm in a non-genomic mechanism only. However, the ESR-mediated signaling pathways are still poorly understood. In this study, we obtained 119 samples from male participants of Caucasian descent who donated semen for standard analysis. We analyzed gene expression of estrogen receptors (ESR1 and ESR2) and their coregulators—proline-, glutamic acid-, and leucine-rich protein 1 (PELP1), and cellular kinase c-Src (SRC). RNA level was established using reverse-transcribed RNA as a template, followed by a polymerase chain reaction. Proteins’ presence was confirmed by western blot and immunocytochemistry techniques. “Normal” values of semen parameters were defined as follows: > 32% sperm with progressive motility, > 4% sperm cells with normal morphology, > 15 × 106 sperm per mL, > 58% live spermatozoa and leukocyte amount < 106 cells per mL, according to WHO 2010 reference. Semen parameters that deviated from these “normal” values were labeled as “abnormal”. Gene expression ratios revealed significant, moderate, and negative correlations for ESR1/ESR2 and weak, negative ESR2/PELP1 correlations in the subgroup of patients with abnormal values of semen parameters. In addition, SRC/PELP1 was moderately and positively correlated in the subgroup with parameters within the reference values established by WHO 2010. Our study showed that both PELP1 scaffolding protein and SRC kinase might influence semen quality via ESRs. It seems that not the expression of a single gene may affect the sperm quality, but more gene-to-gene mutual ratio. Characterization of estrogen-signaling pathway-related genes’ modulated expression in sperm cells could aid in better understanding sperm biology and quality.

Development of pretreatment method for analysis of fungicides and hormones in seawater using LC-MS/MS

A simultaneous analytical method for two prohibited fungicides and their metabolites was developed alongside a simultaneous analytical method for two hormone compounds. For the fungicides, conditions such as the SPE cartridge type, pH conditions, and additives were varied while for the hormones, the elution solvents and pH conditions were varied, and finally, the optimal conditions were selected for each compound group. Target fungicide compounds and medroxyprogesterone acetate were ionized in positive mode while diethylstilbestrol was ionized in negative mode. For the fungicides, with the injection of additives and buffer, the MCX cartridge showed more stable recoveries (85.1-140.0%) compared to the HLB (82.3-251.7%) and combined cartridges (56.1-277.2%). The accuracy and precision of the analytical method were 83.2-130.1% and 6.8-15.0%, respectively. The method detection limit and limit of quantification from this study were lower than those reported in previous studies. Meanwhile, for hormones, methanol was the most appropriate elution solvent in the presence of ascorbic acid additive and at pH 2.5. Accuracy and precision ranged from 75.3-106.6% and 4.3-9.1%, respectively. This study's optimized simultaneous analytical methods can be used to proactively detect prohibited substances in seawater and aquaculture farms.