Prohormone Research Articles

Neurosteroid Metabolites of Gonadal Steroid Hormones in Neuroprotection: Implications for Sex Differences in Neurodegenerative Disease.

Gonadal steroid hormones are neurotrophic and neuroprotective. These effects are modulated by local metabolism of the hormones within the brain. Such control is necessary to maintain normal function, as several signaling pathways that are activated by gonadal steroid hormones in the brain can also become dysregulated in disease. Metabolites of the gonadal steroid hormones—particularly 3α-hydroxy, 5α-reduced neurosteroids—are synthesized in the brain and can act through different mechanisms from their parent steroids. These metabolites may provide a mechanism for modulating the responses to their precursor hormones, thereby providing a regulatory influence on cellular responses. In addition, there is evidence that the 3α-hydroxy, 5α-reduced neurosteroids are neuroprotective in their own right, and therefore may contribute to the overall protection conferred by their precursors. In this review article, the rapidly growing body of evidence supporting a neuroprotective role for this class of neurosteroids will be considered, including a discussion of potential mechanisms that may be involved. In addition, we explore the hypothesis that differences between males and females in local neurosteroid production may contribute to sex differences in the development of neurodegenerative disease.

POMC: The Physiological Power of Hormone Processing.

Pro-opiomelanocortin (POMC) is the archetypal polypeptide precursor of hormones and neuropeptides. In this review, we examine the variability in the individual peptides produced in different tissues and the impact of the simultaneous presence of their precursors or fragments. We also discuss the problems inherent in accurately measuring which of the precursors and their derived peptides are present in biological samples. We address how not being able to measure all the combinations of precursors and fragments quantitatively has affected our understanding of the pathophysiology associated with POMC processing. To understand how different ratios of peptides arise, we describe the role of the pro-hormone convertases (PCs) and their tissue specificities and consider the cellular processing pathways which enable regulated secretion of different peptides that play crucial roles in integrating a range of vital physiological functions. In the pituitary, correct processing of POMC peptides is essential to maintain the hypothalamic-pituitary-adrenal axis, and this processing can be disrupted in POMC-expressing tumors. In hypothalamic neurons expressing POMC, abnormalities in processing critically impact on the regulation of appetite, energy homeostasis, and body composition. More work is needed to understand whether expression of the POMC gene in a tissue equates to release of bioactive peptides. We suggest that this comprehensive view of POMC processing, with a focus on gaining a better understanding of the combination of peptides produced and their relative bioactivity, is a necessity for all involved in studying this fascinating physiological regulatory phenomenon.

Effects of Dietary β-Carotene on the Melanization Response and Growth Rate of Trichoplusia ni (Lepidoptera: Noctuidae).

Animals rely on carotenoids as fundamental precursors for hormones and antioxidants, and animals must acquire carotenoids from their diet. Previous research has shown that insects often absorb carotenoids in amounts proportional to those in their diet, and that carotenoids play key roles in multitrophic interactions. The consumption of diets that provide high levels of antioxidant compounds is associated with high levels of immune responses; however, it is unknown whether individual carotenoids directly influence immune response. Here, the objective of this study was to examine the effect of the carotenoid β-carotene on melanization, a measure of immune response, and growth rate of Trichoplusia ni Hübner (Lepidoptera: Noctuidae). To fulfill the objective, a low, medium, and high concentration of β-carotene, representing the range found in typical host plants, were mixed in an artificial diet, and immune response and growth rate were assessed in fifth instar larvae. Immune response was induced by injection of chromatography beads in to the abdomen of the larvae, and percent melanization was measured after injection. Melanization was greatest when larvae were reared on high β-carotene diets. Mass was measured at 5 and 10 d to assess growth rate. Larvae reared on high β-carotene diets initially gained little mass, but after 10 d larvae reared on no and high β-carotene diets were larger than those reared on other diets. This research has shown that β-carotene has the potential to influence the immune response and growth rate of T. ni.

150. Uteroplacental acute atherosis, cardiovascular biomarkers and vasoactive hormones in pregnancy

Introduction Cardiovascular disease (CVD) is the leading cause of death for women. A history of hypertensive disorders of pregnancy increases the risk. Uteroplacental spiral arterial.acute atherosis is a pregnancy-specific lesion resembling early atherosclerosis, frequently found in preeclampsia. The renin-angiotensin-system (RAS) plays a role in atherosclerosis and is dysregulated in preeclampsia. RAS and natriuretic peptides are physiological opponents in the regulation of arterial blood pressure and intravascular volume control. We previously reported increased concentrations of the natriuretic hormone precursor midregional proatrial natriuretic peptide (MR-proANP) in preeclampsia and suggested a cardiac origin. Objective To assess whether uteroplacental acute atherosis correlates with cardiovascular biomarkers and vasoactive hormones in maternal circulation at delivery in preeclamptic and normotensive pregnancies. Methods Third trimester maternal blood samples were analyzed for renin, prorenin and MR-proANP [normotensive pregnancies n = 50, preeclampsia (PE) n = 71]. Decidua basalis tissue for acute atherosis diagnosis (immunohistochemistry) was available for 53 of these pregnancies (normotensives n = 25 and PE n = 28). Statistical analyses: Skewed data were log-transformed. Linear regression and Fisher’s Exact test were applied as appropriate. A p-value Results A negative correlation between renin and MR-proANP concentrations was confirmed for both study groups. The presence of acute atherosis was not associated with maternal levels of circulating renin, prorenin or MR-proANP. Presence of acute atherosis was neither associated with MR-proANP/renin ratio nor maternal renin in the lower 2 quartiles in combination with MR-proANP in the upper 2 quartiles (normotensives p = 1.0 and PE p = 0.7). Discussion This is the first report on RAS and vasoactive hormones in pregnancy in relation to uteroplacental acute atherosis. We find that maternal circulating biomarkers previously shown to indicate hemodynamic stress in preeclampsia are not increased in pregnancies with acute atherosis. Further studies exploring the longitudinal association between maternal circulating cardiovascular biomarkers and postpartum CVD surrogate markers are underway.

Reduction in voluntary food intake, but not fasting, stimulates hypothalamic gonadotropin-inhibitory hormone precursor mRNA expression in chicks under heat stress

Heat stress is an issue of rising concern across the globe. Recently, we found that mRNA expression of gonadotropin-inhibitory hormone (GnIH), an orexigenic neuropeptide, was increased in the heat-exposed chick brain when food intake was reduced. The aim of the current study was to examine mRNA expression of GnIH and of the glucocorticoid receptors (GRs) in the hypothalamus as well as the plasma corticosterone (CORT) and metabolites in 14-d-old chicks exposed to a high ambient temperature (HT; 40 ± 1 °C for 1 or 5 h) or a control thermoneutral temperature (CT; 30 ± 1 °C), either with free access to food or fasted. Heat stress caused a voluntary reduction of food intake and reduced plasma triacylglycerol concentration, but increased rectal temperature and plasma CORT and glucose concentrations (P < 0.05). Heat stress also increased (P < 0.05) the expression of diencephalic GnIH mRNA in chicks when they reduced food intake voluntarily, but did not do so under fasting conditions. Although the expression of GR mRNA was not altered as a result of heat stress, its expression was decreased (P < 0.05) in fasted chicks at 5 h in comparison with fed chicks. In addition, the rectal temperature of fasted chicks was lower than that of fed chicks under both CT and HT. In conclusion, voluntary reduction of food intake caused an increase in brain GnIH mRNA expression, plasma CORT, and body temperature in chicks under heat stress. Interestingly, brain GnIH mRNA expression was not induced by heat stress in fasted chicks and was not accompanied by a decrease in rectal temperature. These results suggest that the increased expression of brain GnIH mRNA in chicks under heat stress could be a consequence of a mechanism mediated by the voluntary reduction of food intake, but that it is not a consequence of fasting.

Prescribing in pregnancy

Perfluorooctanoic acid (PFOA) is a persistent organic pollutant, which may possess endocrine disrupting properties. Herein, we investigated the possible mechanism(s) of toxicity and steroidogenesis in mouse Leydig cells. MLTC-1 (mouse Leydig tumour cells) cells were exposed to 0, 50, 100 or 200 μM PFOA for 48 h to ascertain their effects on the nuclear (membrane) receptor responses, steroidogenesis pathway and related regulated gene expression and steroid hormone secretion profiles. Our results reveal that nuclear receptors PXR, SR-B1 and LHR are sensitive to PFOA exposure. PFOA can accumulate in mitochondria and alter cholesterol precursor (fatty acid) mitochondrial transport process-related gene expression and thus inhibit steroid hormone precursor (cholesterol) production. In particular, PFOA exhibits biphasic effects on testosterone and progesterone production at differing levels of exposure. These findings indicate the potential endocrine-related effects of PFOA on steroid hormone secretion in Leydig cells and point to a novel disruption model.

Antiproliferative Activity of Non-Calcemic Vitamin D Analogs on Human Melanoma Lines in Relation to VDR and PDIA3 Receptors.

Vitamin D is a precursor for secosteroidal hormones, which demonstrate pleiotropic biological activities, including the regulation of growth and the differentiation of normal and malignant cells. Our previous studies have indicated that the inhibition of melanoma proliferation by a short side-chain, low calcemic analog of vitamin D—21(OH)pD is not fully dependent on the expression of vitamin D receptor (VDR). We have examined the effects of classic vitamin D metabolites, 1,25(OH)2D3 and 25(OH)D3, and two low calcemic vitamin D analogs, (21(OH)pD and calcipotriol), on proliferation, mRNA expression and vitamin D receptor (VDR) translocation in three human melanoma cell lines: WM98, A375 and SK-MEL-188b (subline b of SK-MEL-188, which lost responsiveness to 1,25(OH)2D3 and became VDR−/−CYP27B1−/−). All tested compounds efficiently inhibited the proliferation of WM98 and A375 melanoma cells except SK-MEL-188b, in which only the short side-chain vitamin D analog—21(OH)pD was effective. Overall, 21(OH)pD was the most potent compound in all three melanoma cell lines in the study. The lack of responsiveness of SK-MEL-188b to 1,25(OH)2D3, 25(OH)D3 and calcipotriol is explained by a lack of characteristic transcripts for the VDR, its splicing variants as well as for vitamin D-activating enzyme CYP27B1. On the other hand, the expression of VDR and its splicing variants and other vitamin D related genes (RXR, PDIA3, CYP3A4, CYP2R1, CYP27B1, CYP24A1 and CYP11A1) was detected in WM98 and A375 melanomas with the transcript levels being modulated by vitamin D analogs. The expression of VDR isoforms in WM98 cells was stimulated strongly by calcipotriol. The antiproliferative activities of 21(OH)pD appear not to require VDR translocation to the nucleus, which explains the high efficacy of this noncalcemic pregnacalciferol analog in SK-MEL-188b melanoma, that is, VDR−/−. Therefore, we propose that 21(OH)pD is a good candidate for melanoma therapy, although the mechanism of its action remains to be defined.

An automated screening method for detecting compounds with goitrogenic activity using transgenic zebrafish embryos.

The knowledge on environmentally relevant chemicals that may interfere with thyroid signaling is scarce. Here, we present a method for the screening of goitrogens, compounds that disrupt the thyroid gland function, based on the automatic orientation of zebrafish in a glass capillary and a subsequent imaging of reporter gene fluorescence in the thyroid gland of embryos of the transgenic zebrafish line tg(tg:mCherry). The tg(tg:mCherry) reporter gene indicates a compensatory upregulation of thyroglobulin, the thyroid hormone precursor, in response to inhibition of thyroid hormone synthesis. Fish embryos were exposed to a negative control compound (3,4-dichloroaniline), or a concentration series of known goitrogenic compounds (resorcinol, methimazole, potassium perchlorate, 6-propyl-2-thiouracil, ethylenethiourea, phloroglucinol, pyrazole) with maximum exposure concentration selected based on mortality and/or solubility. Exposure to 3,4-dichloroaniline decreased the fluorescence signal. All goitrogenic compounds exhibited clear concentration-dependent inductions of reporter fluorescence 1.4 to 2.6 fold above control levels. Concentration-response modelling was used to calculate goitrogenic potencies based on EC50 values. The new automated method offers an efficient screening approach for goitrogenic activity.

Plasma membrane cholesterol trafficking in steroidogenesis.

Cholesterol is an important component of plasma membranes (PMs) and the precursor of all steroid hormones. In steroidogenic tissues, upon hormone stimulation, there is a rapid transfer of cholesterol to the mitochondria, which is the site of the initial step in steroidogenesis. In the current study, we examined PM cholesterol trafficking for steroidogenesis. In a mitochondrial reconstitution assay, adrenal PMs supported steroidogenesis in the absence of additional transport proteins. Depletion of cholesterol in PMs by 50% eliminated the membranes' ability to support steroidogenesis in vitro and reduced steroid production in intact Y1 adrenocortical cells. Syntaxin (STX)-5 and α-soluble N-ethylmaleimide-sensitive factor attachment protein (α-SNAP) are enriched in adrenal PMs, and adrenocorticotropic hormone treatment of rats recruited STX5 and α-SNAP to adrenal PMs and mitochondria. Immunodepletion of STX5 and α-SNAP from PMs decreased steroidogenesis supported by PMs in vitro. Protease digestion of PMs decreased, whereas recombinant STX5 or α-SNAP restored, the PMs' ability to support steroidogenesis. Knockdown of either STX5 or α-SNAP in Y1 cells decreased stimulated steroidogenesis. These results indicate that STX5 and α-SNAP facilitate cholesterol trafficking from PMs to mitochondria for adrenal steroid synthesis and underscore the importance of vesicular trafficking of PM cholesterol for steroidogenesis.-Deng, B., Shen, W.-J., Dong, D., Azhar, S., Kraemer, F. B. Plasma membrane cholesterol trafficking in steroidogenesis.

Plant Sources, Extraction Methods, and Uses of Squalene

Squalene (SQ) is a natural compound, a precursor of various hormones in animals and sterols in plants. It is considered a molecule with pharmacological, cosmetic, and nutritional potential. Scientific research has shown that SQ reduces skin damage by UV radiation, LDL levels, and cholesterol in the blood, prevents the suffering of cardiovascular diseases, and has antitumor and anticancer effects against ovarian, breast, lung, and colon cancer. The inclusion of SQ in the human diet is recommended without causing health risks; however, its intake is low due to the lack of natural sources of SQ and efficient extraction methods which limit its commercialization. Biotechnological advances have developed synthetic techniques to produce SQ; nevertheless, yields achieved are not sufficient for global demand for industrial or food supplement purposes. The effect on the human body is one of the scientific issues still to be addressed; few research studies have been developed with SQ from seed or vegetable sources to use it in the food sector even though squalene was discovered more than half a century ago. The aim of this review is to provide an overview of SQ to establish research focus with special reference to plant sources, extraction methods, and uses.

Absence of a correlation between intra-follicular androgen and AMH levels and the oocyte count in poor responder patients undergoing in vitro fertilization

Androgens are male hormones and precursors of female hormones. Testosterone (T), dehydroepiandrostenedione (DHEA), and anti-mullerian hormone (AMH) enhance follicular recruitment and promote follicular growth and development. Clinical reports have demonstrated that co-treatment with androgens, such as DHEA and Androderm (transdermal testosterone) may increase both quantity and quality of oocytes and embryos and improve pregnancy outcomes in women with diminished ovarian function or premature ovarian failure. This study was designed to find correlations between intra-follicular androgen and AMH levels and the amount of oocytes retrieved from ovarian responders and poor-responders. We enrolled 40 patients undergoing in vitro fertilization (IVF) and tested their follicular fluid for testosterone, DHEA, and AMH levels. AMH levels were significantly different between the groups. However, no correlations were observed between the levels of testosterone, DHEA, or AMH and the amounts of retrieved oocytes. No correlations were observed between androgens and AMH levels with the amount of oocytes in either group. Thus, there is no correlation between intra-follicular androgen and AMH levels and the amount of oocytes in both responders and poor responders. A larger sample size and multi-center study design is needed to confirm this.

Resveratrol Inhibits Key Steps of Steroid Metabolism in a Human Estrogen-Receptor Positive Breast Cancer Model: Impact on Cellular Proliferation.

The role of resveratrol (RES) in preventing breast cancer is controversial, as low concentrations may stimulate the proliferation of estrogen-receptor alpha positive (ERα+) breast cancer cells. As metabolism is the key factor in altering cellular estrogens, thereby influencing breast tumor growth, we investigated the effects of RES on the formation of estrogen metabolites, namely 4-androstene-3,17-dione (AD), dehydroepiandrosterone (DHEA), dehydroepiandrosterone-3-O-sulfate (DHEA-S), estrone (E1), estrone-3-sulfate (E1-S), 17β-estradiol (E2), 17β-estradiol-3-O-(β-D-glucuronide) (E2-G), 17β-estradiol-3-O-sulfate (E2-S), 16α-hydroxy-17β-estradiol (estriol, E3), and testosterone (T) in ERα- MDA-MB-231 and ERα+ MCF-7 cells. Incubation of both of the cell lines with the hormone precursors DHEA and E1 revealed that sulfation and glucuronidation were preferred metabolic pathways for DHEA, E1 and E2 in MCF-7 cells, compared with in MDA-MB-231 cells, as the Vmax values were significantly higher (DHEA-S: 2873.0 ± 327.4 fmol/106 cells/h, E1-S: 30.4 ± 2.5 fmol/106 cells/h, E2-S: 24.7 ± 4.9 fmol/106 cells/h, E2-G: 7.29 ± 1.36 fmol/106 cells/h). RES therefore significantly inhibited DHEA-S, E1-S, E2-S and E2-G formation in MCF-7, but not in MDA-MB-231 cells (Kis: E2-S, 0.73 ± 0.07 μM < E1-S, 0.94 ± 0.03 μM < E2-G, 7.92 ± 0.24 μM < DHEA-S, 13.2 ± 0.2 μM). Suppression of these metabolites subsequently revealed twofold higher levels of active E2, concomitant with an almost twofold increase in MCF-7 cell proliferation, which was the most pronounced upon the addition of 5 μM RES. As the content of RES in food is relatively low, an increased risk of breast cancer progression in women is likely to only be observed following the continuous consumption of high-dose RES supplements. Further long-term human studies simultaneously monitoring free estrogens and their conjugates are therefore highly warranted to evaluate the efficacy and safety of RES supplementation, particularly in patients diagnosed with ERα+ breast cancer.

A rapid LC-MS method for qualitative and quantitative profiling of plant apocarotenoids

Carotenoid cleavage products (apocarotenoids; APOs) exert important biological functions in light perception and as vitamin A source, signaling molecules, hormone precursors, pigments and volatiles. However, an analytical method that allows simultaneous profiling of these diverse compounds is still missing. We developed an efficient method to analyze APOs present in plant tissues, which is based on ultra-high performance liquid chromatographic separation and high-resolution hybrid quadrupole-Orbitrap (Q-Orbitrap) mass spectrometry (MS). Our approach allowed unambiguous identification and quantification of volatile and non-volatile APOs in a single run. Modified sample preparation and optimized ultra-high performance liquid chromatography (UHPLC)-MS parameters permitted the measurement of APOs in Oryza sativa seedlings and Spinacia oleracea leaves, unraveling 20 endogenous APOs with chain lengths ranging from C10 to C30, confirmed by high-resolution MS, MS/MS data and using synthetic standards. Our experimentation demonstrates that the usage of methanol with 0.1% butylated hydroxytoluene facilitates the extraction of both short-chain and long-chain APOs from plant materials. In addition, our validated analytical method allows the quantitative analysis of APOs with a wide content range from 2.5 pg/mg to 10 ng/mg dried weight. The adoption of the analytical protocol, as described in this study, realizes the measurement of volatile APOs by using a LC-MS method, hence, allowing informative and reliable profiling of APOs, which is important for determining the content of these compounds in food and crucial for understanding their function and metabolism in plants.

Precursors of neuropeptides and peptide hormones in the genomes of tardigrades

Tardigrades are a key group for understanding the evolution of the Ecdysozoa, a large clade of molting animals that also includes arthropods and nematodes. However, little is known about most aspects of their basic biology. Neuropeptide and peptide hormone signaling has been extensively studied in arthropods and nematodes (particularly regarding their roles in molting in arthropods), but very little is known about neuropeptide signaling in other ecdysozoans. In this work, different strategies were used to search for neuropeptide and peptide hormone precursors in the genomes of the tardigrades Hypsibius dujardini and Ramazzottius varieornatus. In general, there is a remarkable similarity in the complement of neuropeptides and their sequences between tardigrades and arthropods. The precursors found in tardigrades included homologs of achatin, allatostatins A, B and C, allatotropin, calcitonin, CCHamide, CCRFa, corazonin, crustacean cardioactive peptide, diuretic hormone 31, diuretic hormone 44, ecdysis triggering hormone, eclosion hormone, gonadotropin-releasing hormone (GnRH), GSEFLamide, insulin-like peptides, ion transport peptide, kinin, neuropeptide F, orcokinin, pigment dispersing hormone, proctolin, pyrokinin, RYamide, short neuropeptide F, sulfakinin, tachykinin, trissin and vasopressin. In most cases, homologs of known cognate receptors for each neuropeptide family could only be identified when the precursors were also present in the genome, further supporting their identification. Some neuropeptide precursor genes have undergone several duplications in tardigrades, including allatostatin A and C, corazonin, GnRH, eclosion hormone, sulfakinin and trissin. Furthermore, four novel families of candidate neuropeptide precursors were identified (two of which could also be found in several arthropods). To the best of my knowledge, this work represents the first genome-wide search for neuropeptide precursors in any ecdysozoan species outside arthropods and nematodes, and is a necessary first step towards understanding neuropeptide function in tardigrades.

The role of steroid metabolome analysis for the diagnosis and follow-up of adrenocortical tumors.

The diagnostic work-up of adrenal tumors, often incidentally discovered, has emerged as an ever-increasing diagnostic problem for clinical endocrinologists. No imaging modality has sufficiently high sensitivity and specificity at differentiating benign from malignant adrenal lesions. It has long been observed that adrenocortical carcinomas (ACCs) present an immature pattern of steroidogenesis, dominated by steroid hormone precursors. Modern mass spectrometry-based assays can generate multi-steroid metabolite profiles in urine collections, which can detect differences between ACCs and benign adrenocortical adenomas (ACAs). This review summarizes the promising results of studies which have applied steroid metabolite profiling in biological fluids as a novel diagnostic tool for patients with adrenal tumors, as well as the challenges and limitations of this approach. It also discusses the potential role of steroid profiling as a biochemical surveillance tool to detect recurrence in patients who have undergone resection of an ACC.

A synthetic diosgenin primary amine derivative attenuates LPS-stimulated inflammation via inhibition of NF-κB and JNK MAPK signaling in microglial BV2 cells

Diosgenin, a precursor of steroid hormones in plants, is known to exhibit diverse pharmacological activities including anti-inflammatory properties. In this study, (3β, 25R)‑spirost‑5‑en‑3‑oxyl (2‑((2((2‑aminoethyl)amino)ethyl)amino)ethyl) carbamate (DGP), a new synthetic diosgenin derivative incorporating primary amine was used to investigate its anti-inflammatory effects and underlying mechanisms of action in lipopolysaccharide (LPS)-stimulated microglial BV2 cells. Pretreatment with DGP resulted in significant inhibition of nitric oxide (NO) synthesis, and down-regulation of nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in LPS-stimulated microglial BV2 cells. In addition, DGP decreased the production of reactive oxygen species (ROS) and pro-inflammatory cytokines such as interleukin (IL)-6, IL-1β, and tumor necrosis factor alpha (TNF-α). The inhibitory effects of DGP on these inflammatory mediators in LPS-stimulated microglial BV2 cells were regulated by NF-κB signaling through blocking p65 nuclear translocation and NF-κB p65/DNA binding activity. DGP also blocked the phosphorylation of c-Jun amino-terminal kinase (JNK), but not p38 kinase or extracellular signal-regulated kinases (ERK). The NF-κB inhibitor JSH-23 and JNK-specific inhibitor SP600125 significantly decreased NO production and IL-6 release in LPS-stimulated BV2 cells, respectively. The overall results demonstrate that DGP has anti-inflammatory effects on LPS-stimulated BV2 cells via inhibition of NF-κB and JNK activation, suggesting that DGP is a potential prophylactic agent in various neurodegenerative disorders.

Neuroendocrine mechanism of food intake and energy regulation in Japanese quail under differential simulated photoperiodic conditions: Involvement of hypothalamic neuropeptides, AMPK, insulin and adiponectin receptors

Neuroendocrine coordination between the reproductive and energy regulatory hypothalamic circuitries not only tightly regulates food intake and energy expenditure but also maintains the body weight and reproduction. The effect of different simulated photoperiodic conditions on food intake and neuroendocrine mechanism of energy homeostasis in Japanese quail is not investigated till date. Hence, our present study is designed to elucidate the effect of different simulated photoperiodic conditions on food consumption and neuroendocrine mechanism(s) of energy regulation in this poultry species. The alterations in hypothalamic energy balancing neuropeptides (NPY/AgRP/CART), polypeptide hormone precursor (POMC), protein kinase (AMPK-p-AMPK) as well as receptors of insulin and adiponectin [Insulin Receptor (IR), Adiponectin Receptor 1 & 2] have been investigated in photosensitive (PS), scotorefractory (SR),photorefractory (PR) and scotosensitive (SS) quail. Immunofluorescence and western blotting were used to quantify the expression of these peptides and proteins. Results showed increased food consumption and body weight gain, along with increased expression of NPY, AgRP, IR, adiponectin receptors and p-AMPK, decreased CART and POMC in the hypothalamus of photosensitive and scotorefractory quail. While, opposite findings were observed in photorefractory and scotosensitive quail. Hence, this study may suggest the hypothalamic energy channelization towards reproductive axis in photosensitive and scotorefractory quail to support the full breeding conditions, while hypothalamic energy deprivation in photorefractory and scotosensitive quail leads to reproductive quiescence.

Prediction of Molecular Mechanisms for LianXia NingXin Formula: A Network Pharmacology Study.

Objectives: Network pharmacological methods were used to investigate the underlying molecular mechanisms of LianXia NingXin (LXNX) formula, a Chinese prescription, to treat coronary heart disease (CHD) and disease phenotypes (CHD related diseases and symptoms).Methods: The different seed gene lists associated with the herbs of LXNX formula, the CHD co-morbid diseases and symptoms which were relieved by the LXNX formula (co-morbid diseases and symptoms) were curated manually from biomedical databases and published biomedical literatures. Module enrichment analysis was used to identify CHD-related disease modules in the protein–protein interaction (PPI) network which were also associated to the targets of LXNX formula (LXNX formula’s CHD modules). The molecular characteristics of LXNX formula’s CHD modules were investigated via functional enrichment analysis in terms of gene ontology and pathways. We performed shortest path analysis to explore the interactions between the drug targets of LXNX formula and CHD related disease phenotypes (e.g., co-morbid diseases and symptoms).Results: We identified two significant CHD related disease modules (i.e., M146 and M203), which were targeted by the herbs of LXNX formula. Pathway and GO term functional analysis results indicated that G-protein coupled receptor signaling pathways (GPCR) of M146 and cellular protein metabolic process of M203 are important functional pathways for the respective module. This is further confirmed by the shortest path analysis between the drug targets of LXNX formula and the aforementioned disease modules. In addition, corticotropin releasing hormone (CRH) and natriuretic peptide precursor A (NPPA) are the only two LXNX formula target proteins with the low shortest path length (on average shorter than 3) to their respective CHD module and co-morbid disease and symptom gene groups.Conclusion: G-protein coupled receptor signaling pathway and cellular protein metabolic process are the key LXNX formula’s pathways to treat CHD disease phenotypes, in which CRH and NPPA are the two key drug targets of LXNX formula. Further evidences from Chinese herb pharmacological databases indicate that Pinellia ternata (Banxia) has relatively strong adjustive functions on the two key targets.

Nutritional quality of fish faeces is enhanced by highly unsaturated fatty acid-producing heterotrophic protozoa

Highly unsaturated fatty acids such as 20:5n3 (EPA) are both hormone precursors and cell membrane components, making them important nutrients for aquatic animals. Many animals must obtain EPA from their diets because they cannot synthesize enough EPA to meet their requirements, and algae are the main source of EPA in aquatic ecosystems. In a previous study, we detected EPA in the faeces of Danio rerio, a freshwater fish, even though the fish consumed a green algae diet that did not contain EPA. The objective of this study was to determine why EPA was detected in fish faeces. A significant positive relationship was detected between the number of heterotrophic protozoa and the concentration of EPA in the faeces, which suggests that this EPA was of protozoan origin. In addition, another experiment showed that protozoa adhered to faeces far more than the green algal diet remnants, which indicates that protozoa preferred to swarm on faeces. Furthermore, we cultured protozoa in an EPA-free medium and fed them a bacterial diet also lacking EPA, and found that Cyclidium sp. synthesized EPA de novo. The results demonstrate that protozoa produce essential fatty acids and enhance the nutritional quality of animal faeces in detritus-based food webs in freshwater ecosystems.

Transcriptomic and Neuropeptidomic Analysis of the Stick Insect, Carausius morosus.

One of the most thoroughly studied insect species, with respect to locomotion behavior, is the stick insect Carausius morosus. Although detailed information exists on premotor networks controlling walking, surprisingly little is known about neuropeptides, which are certainly involved in motor activity generation and modulation. So far, only few neuropeptides were identified from C.morosus or related stick insects. We performed a transcriptome analysis of the central nervous system to assemble and identify 65 neuropeptide and protein hormone precursors of C.morosus, including five novel putative neuropeptide precursors without clear homology to known neuropeptide precursors of other insects ( Carausius neuropeptide-like precursor 1, HanSolin, PK-like1, PK-like2, RFLamide). Using Q Exactive Orbitrap and MALDI-TOF mass spectrometry, 277 peptides including 153 likely bioactive mature neuropeptides were confirmed. Peptidomics yielded a complete coverage for many of the neuropeptide propeptides and confirmed a surprisingly high number of heterozygous sequences. Few neuropeptide precursors commonly occurring in insects, including those of insect kinins and sulfakinins, could neither be found in the transcriptome data nor did peptidomics support their presence. The results of our study represent one of the most comprehensive peptidomic analyses on insects and provide the necessary input for subsequent experiments revealing neuropeptide function in greater detail.