Beyond transport: cytokinin ribosides are translocated and active in regulating the development and environmental responses of plants.

MAIN CONCLUSION: Riboside type cytokinins are key components in cytokinin metabolism, transport, and sensitivity, making them important functional signals in plant growth and development and environmental stress responses. Cytokinin (CKs) are phytohormones that regulate multiple processes in plants and are critical for agronomy, as they are involved in seed filling and plant responses to biotic and abiotic stress. Among the over 30 identified CKs, there is uncertainty about the roles of many of the individual CK structural forms. Cytokinin free bases (CKFBs), have been studied in great detail, but, by comparison, roles of riboside-type CKs (CKRs) in CK metabolism and associated signaling pathways and their distal impacts on plant physiology remain largely unknown. Here, recent findings on CKR abundance, transport and localization, are summarized, and their importance in planta is discussed. The history of CKR analyses is reviewed, in the context of the determination of CK metabolic pathways, and research on CKR affinity for CK receptors, all of which yield essential insights into their functions. Recent studies suggest that CKR forms are a lot more than a group of transport CKs and, beyond this, they play important roles in plant development and responses to environmental stress. In this context, this review discusses the involvement of CKRs in plant development, and highlight the less anticipated functions of CKRs in abiotic stress tolerance. Based on this, possible mechanisms for CKR modes of action are proposed and experimental approaches to further uncover their roles and future biotechnological applications are suggested.

cAMP Biosensor Assay Using BacMam Expression System: Studying the Downstream Signaling of LH/hCG Receptor Activation.

Cyclic adenosine monophosphate (cAMP) serves as a second messenger for numerous G-protein-coupled receptors. Changes in cellular cAMP levels reflect the biological activity of various GPCR-specific agents, including protein hormones. cAMP biosensors based on detection of Förster-type resonance energy transfer (FRET) offer unique advantages including the ratiometric nature of measurement, adjustable affinity toward detected molecule, capability of monitoring kinetics of cAMP release, and compatibility with the multi-well format and fluorescence plate reader platforms. In this chapter, we introduce the optimized version of the previously reported method to achieve sufficient and reproducible level of cAMP biosensor protein expression with the means of BacMam transduction system. As a practical challenge, we address the applicability of the designed assay for screening of biological activity of human hormones, including human chorionic gonadotropin (hCG) bearing different posttranslational modifications.

Transcriptome Analysis of Female and Male Conopomorpha sinensis (Lepidoptera: Gracilariidae) Adults With a Focus on Hormone and Reproduction.

Conopomorpha sinensis Bradley is the dominant borer pest of litchi and longan in the Asian-pacific area. Reduction or interference of reproduction and mating of adult moths is one of the most used strategies to control C. sinensis. Insect reproduction is a critical biological process closely related to endocrine control. Conopomorpha sinensis genome and transcriptome information is limited, hampering both our understanding of the molecular mechanisms underlying hormone activity and reproduction and the development of control strategies for this borer pest. To explore the sex differences in gene expression profiles influencing these biological processes, de novo transcriptomes were assembled from female and male adult C. sinensis specimens. This analysis yielded 184,422 unigenes with an average length of 903 bp and 405,961 transcripts after sequencing and assembly. About 45.06, 22.41, 19.53, 34.05, 35.82, 36.42, and 19.85% of the unigenes had significant matches in seven public databases. Subsequently, gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis revealed comprehensive information about the function of each gene and identified enriched categories and pathways that were associated with the 2,890 female-biased genes and 2,964 male-biased genes. In addition, we identified some important unigenes related to hormone activity and reproduction among the sex-differentially expressed genes (DEGs), including unigenes coding for ecdysone-induced protein 78C, juvenile hormone (JH)-regulated gene fatty acyl-CoA reductase, vitellogenin, etc. Our findings provide a more comprehensive portrait of the sex differences involved in the relationship of two important physiological features-hormone activity and reproduction in C. sinensis and members of the family Gracillariidae.

[Adrenal incidentaloma : Diagnostic and therapeutic concept from an endocrinological perspective]. / Inzidentalome der Nebennieren : Diagnostisches und therapeutisches Konzept aus endokrinologischer Sicht.

An adrenal incidentaloma is an adrenal mass detected on imaging that was not performed for suspected adrenal disease. The prevalence is approximately 3% and increases up to 10% in older people. The risk of malignancy and a hormone excess have to be evaluated. Approximately 15% of incidentalomas harbor an overproduction of hormones, in particular primary aldosteronism (Conn's syndrome), hypercortisolism (Cushing's syndrome) and pheochromocytoma. Primary aldosteronism is the main cause of endocrine hypertension. It is characterized by an overproduction of aldosterone usually due to a unilateral adenoma or an idiopathic, often bilateral hyperplasia. The aldosterone to renin ratio is an established screening parameter for the diagnosis. If the ratio is elevated a confirmatory test, e. g. saline infusion test, should follow. Usually an adrenal venous catheter has to be used to discriminate between unilateral and bilateral aldosterone overproduction. In the case of unilateral overproduction an adrenalectomy is recommended, otherwise treatment is carried out with an aldosterone antagonist. For the diagnosis of an adrenal Cushing's syndrome a dexamethasone suppression test and a suppressed or in the lower limit of normal ACTH is required. The rare pheochromocytoma is a catecholamine-producing tumor. The diagnosis is carried out by determination of metanephrines in plasma or in 24 h urine samples. Unilateral adrenal tumors leading to clinically significant hormone excess or tumors with suspicion of malignancy should be surgically removed. A minimally invasive adrenalectomy is normally the method of choice in patients with a unilateral adrenal tumor <6 cm and without local tumor invasion. In unilateral, clearly benign, non-functioning, small adrenal tumors (<4 cm) surgery is not required.

Comparison of in vitro hormone activities of novel flame retardants TBB, TBPH and their metabolites TBBA and TBMEPH using reporter gene assays.

The anti-androgenic and anti-thyroid hormonal activities of the two novel brominated flame retardants, TBB and TBPH and of their metabolites TBBA and TBMEPH have been compared using the luciferase reporter gene assays. Only the parent compounds TBB and TBPH exhibited anti-glucocorticoid activity with IC50 values of 1.9 µM and 0.3 µM. Furthermore, mode of action for these two compounds is by direct competing to the glucocorticoid receptor (GR) with IC50 values of 0.03 µM and 0.002 µM. All four tested compounds possess anti-androgenic and anti-thyroid hormonal activities, without agonist activities on the respective receptors. Anti-androgenic activities with IC50 values of 43.5 µM, 0.1 µM, 47.5 µM and 1.3 µM were found for TBB, TBPH, TBBA and TBMEPH. The anti-thyroid hormonal IC50 values of 37.5 µM, 0.1 µM, 22.8 µM and 32.3 µM for TBB, TBPH, TBBA and TBMEPH, together with the above quoted results, indicate that metabolism can modify anti-androgenic, anti-glucocorticoid and anti-thyroid hormonal effects of these novel brominated flame retardants. Furthermore, the parent flame retardants are shown to be able to disrupt the function of the GR as antagonists by direct competition to the receptor.

Assessment of multiple hormone activities of a UV-filter (octocrylene) in zebrafish (Danio rerio).

In this study, zebrafish (Danio rerio) were exposed to a UV-filter-octocrylene (OCT) with elevated concentrations for 28 d. The total body accumulation of OCT in zebrafish was found to reach 2321.01 ("L" level), 31,234.80 ("M" level), and 70,593.38 ng g(-1) ("H" level) when the average OCT exposure concentration was controlled at 28.61, 505.62, and 1248.70 µg L(-1), respectively. Gross and histological observations as well as RT-qPCR analysis were conducted to determine the effects of OCT accumulation on zebrafish. After exposure, the gonad-somatic index and percentage of vitellogenic oocytes were found to increase significantly in the ovaries of female zebrafish at the H accumulation level. Significant up-regulation of esr1 and cyp19b were observed in the gonads, as well as vtg1 in the livers for both female and male zebrafish. At M and H accumulation levels, apparent down-regulation of ar was observed in the ovaries and testis of the female and male zebrafish, respectively. Although the extent of the effects on zebrafish differed at different accumulation levels, the induction of vtg1 and histological changes in the ovaries are indications of estrogenic activity and the inhibition of esr1 and ar showed antiestrogenic and antiandrogenic activity, respectively. Thus, as OCT could easily accumulate in aquatic life such as zebrafish, one of its most of concern hazards would be the disturbance of the histological development and its multiple hormonal activities.

A critical review finds styrene lacks direct endocrine disruptor activity.

The European Commission lists styrene (S) as an endocrine disruptor based primarily on reports of increased prolactin (PRL) levels in S-exposed workers. The US Environmental Protection Agency included S in its list of chemicals to be tested for endocrine activity. Therefore, the database of S for potential endocrine activity is assessed. In vitro and in vivo screening studies, as well as non-guideline and guideline investigations in experimental animals indicate that S is not associated with (anti)estrogenic, (anti)androgenic, or thyroid-modulating activity or with an endocrine activity that may be relevant for the environment. Studies in exposed workers have suggested elevated PRL levels that have been further examined in a series of human and animal investigations. While there is only one definitively known physiological function of PRL, namely stimulation of milk production, many normal stress situations may lead to elevations without any chemical exposure. Animal studies on various aspects of dopamine (DA), the PRL-regulating neurotransmitter, in the central nervous system did not give mechanistic explanations on how S may affect PRL levels. Overall, a neuroendocrine disruption of PRL regulation cannot be deduced from a large experimental database. The effects in workers could not consistently be reproduced in experimental animals and the findings in humans represented acute reversible effects clearly below clinical and pathological levels. Therefore, unspecific acute workplace-related stress is proposed as an alternative mode of action for elevated PRL levels in workers.