Hormone-like Activity Research Articles

Estrogen-like activity of seafood related to environmental chemical contaminants

BackgroundA wide variety of environmental pollutants occur in surface waters, including estuarine and marine waters. Many of these contaminants are recognised as endocrine disrupting chemicals (EDCs) which can adversely affect the male and female reproductive system by binding the estrogen receptor and exhibiting hormone-like activities. In this study the estrogenic activity of extracts of edible marine organisms for human consumption from the Mediterranean Sea was assayed.MethodsMarine organisms were collected in two different areas of the Mediterranean Sea. The estrogenic activity of tissues was assessed using an in vitro yeast reporter gene assay (S. cerevisiae RMY 326 ER-ERE). Concentrations of polychlorinated biphenyls (PCBs) (congeners 28, 52, 101, 118, 138, 153, 180) in fish tissue was also evaluated.ResultsThirty-eight percent of extracts showed a hormone-like activity higher than 10% of the activity elicited by 10 nM 17b-estradiol (E2) used as control.Total PCB concentrations ranged from 0.002 up to 1.785 ng/g wet weight. Chemical analyses detected different levels of contamination among the species collected in the two areas, with the ones collected in the Adriatic Sea showing concentrations significantly higher than those collected in the Tyrrhenian Sea (p < 0.01).ConclusionThe more frequent combination of chemicals in the samples that showed higher estrogenic activity was PCB 28, PCB 101, PCB 153, PCB 180.The content of PCBs and estrogenic activity did not reveal any significant correlation.

Role of Resistin-Like Molecule (RELM)-β in Allergic Airway Inflammation

RATIONALE: RELM-β, a novel protein with both cytokine and hormone-like activities is produced under Th2-type immune regulation in the gut in mice. We hypothesized that this molecule is also involved in the pathogenesis of the allergic airway response.

Relationship between fatty acids and the endocrine and neuroendocrine system

Significant interactions exist between fatty acids and the endocrine system. Dietary fatty acids alter both hormone and neuropeptide concentrations and also their receptors. In addition, hormones affect the metabolism of fatty acids and the fatty acid composition of tissue lipids. The principal hormones involved in lipid metabolism are insulin, glucagon, catecholamines, cortisol and growth hormone. The concentrations of these hormones are altered in chronic degenerative conditions such as diabetes and cardiovascular disease, which in turn leads to alterations in tissue lipids. Lipogenesis and lipolysis, which modulate fatty acid concentrations in plasma and tissues, are under hormonal control. Neuropeptides are also involved in lipid metabolism in brain and other tissues. Polyunsaturated fatty acids are also precursors for eicosanoids including prostaglandins, leucotrienes, and thromboxanes, which have hormone-like activities. Fatty acids in turn affect the endocrine system. Saturated and trans fatty acids decrease insulin concentration leading to insulin resistance. In contrast, polyunsaturated fatty acids increase plasma insulin concentration and decrease insulin resistance. In humans, ω3 polyunsaturated fatty acids alter the levels of opioid peptides in plasma. Free fatty acids have been reported to inhibit glucagon release. Fatty acids also affect receptors for hormones and neuropeptides.

Endocrine disruptor contaminants in water and their adverse effects in humans

Since the early 1990s the occurrence of pharmaceuticals and chemicals with hormone and hormone-like activity in the environment, has received considerable attention. There are approximately 150 distinct substances being suspected to act as endocrine disruptors. Anumber of endocrine disruptors have been detected in ground and drinking water. Endocrine disrupting compounds are of particular concern as these cannot only interfere with the natural production and metabolism of hormones in the body, but also represent a serious carcinogenic risk. The hormone-like action of drinking water contaminants could affect the proliferation and cell cycle of cells expressing hormone receptors. Moreover, hormone-like agents may modulate the susceptibility of cancer cells to chemotherapy.

Bromophenols, both present in marine organisms and in industrial flame retardants, disturb cellular Ca 2+ signaling in neuroendocrine cells (PC12)

Bromophenols are present in polychaetes as well as in algae in marine environments including the North Sea. They are thought to cause the typical sea-like taste and flavour. The ecological function of brominated phenols is not clear yet, but they may play a role in chemical defence and deterrence [Kicklighter, C.E., Kubaneck, J., Hay, M.E., 2004. Do brominated natural products defend marine worms from consumers? Some do, most don’t. Limnol. Oceanogr. 49, 430–441]. Some brominated phenols are commercially used as industrial flame retardants as, e.g., 2,4,6-tribromophenol and are suspected to disrupt the humoral system by showing tyroid hormone-like activity [Legler, I., Brouwer, A., 2003. Are brominated flame retardants endocrine disruptors? Environ. Int. 29, 879–885]. In this study 2-bromophenol (2-BP), 4-bromophenol (4-BP), 2,4-dibromophenol (2,4-DBP), 2,6-dibromophenol (2,6-DBP) and 2,4,6-tribromophenol (2,4,6-TBP), all of which are present in marine organisms, were tested. Especially 2,4-DBP and 2,4,6-TBP showed a significant effect on the Ca 2+ homeostasis in endocrine cells (PC 12). The reduction of depolarization induced Ca 2+ elevations by 2,4-DBP and 2,4,6-TBP and the increase of intracellular Ca 2+ by both substances, partly released from intracellular stores, may suggest a link to the disrupting effect of endocrine systems by brominated phenols. 2,4-DBP was the most potent substance we tested in respect to inhibition of voltage dependent Ca 2+ currents as revealed in whole cell patch clamp experiments. Brominated phenols disturb cellular Ca 2+ signaling with differential efficacy, depending on the number and position of bromine.

T-screen to quantify functional potentiating, antagonistic and thyroid hormone-like activities of poly halogenated aromatic hydrocarbons (PHAHs)

The present study investigates chemical thyroid hormone disruption at the level of thyroid hormone receptor (TR) functioning. To this end the (ant)agonistic action of a series of xenobiotics was tested in the newly developed T-screen. This assay makes use of a GH 3 rat pituitary cell line, that specifically proliferates when exposed to 3,3′,5-triiodo- l-thyronine (T 3). The growth stimulatory effect is mediated via T 3-receptors. (Ant)agonistic and potentiating action of compounds was studied in absence and presence of T 3 at its EC 50 level (0.25 nM). The compounds tested included the specific TR-antagonist amiodarone, as well as a series of brominated diphenyl ethers (BDEs), including specifically synthesized BDEs with a structural resemblance to 3,5-diiodo- l-thyronine (T 2), T 3 and T 4 (3,3′,5,5′-tetraiodo- l-thyronine). The results obtained reveal that only BDE206 and amiodarone are specific antagonists. Interestingly some compounds which did not respond in the T-screen in absence of T 3, potentiated effects when tested in combination with T 3. This points at possibilities for disruption at the TR in vivo, where exposure generally occurs in presence of T 3. Altogether the results of the present study show that the newly developed T-screen can be used as a valuable tool for identification and quantification of compounds active in disturbing thyroid hormone homeostasis at the level of TR-functioning.

A New Frontier in Soy Bioactive Peptides that May Prevent Age-related Chronic Diseases.

During gastrointestinal digestion or food processing of proteins, small peptides can be released and may act as regulatory compounds with hormone-like activities. Numerous biologically active peptides (bioactive peptides) have been identified. Most bioactive peptides are derived from milk and dairy products, with the most common being angiotensin converting enzyme inhibitory peptides. Soybean protein and soybean derived peptides also play an important role in soybean physiological activities, particularly those related to the prevention of chronic diseases. However, the bioactive potential of soybean derived bioactive peptides is yet to be fully appreciated. After a general introduction of approaches and advances in bioactive peptides from food sources, this review focuses on bioactive peptides derived from soybean proteins and their physiological properties. Technological approaches to generate bioactive peptides, their isolation, purification, characterization, and quantification, and further application in food and drug design are also presented. Safety concerns, such as potential toxicity, allergenicity, and sensory aspect of these peptides are likewise discussed.

Teratogenic effects of retinoic acid on neurulation in mice embryos

Retinoic acids (RA) are natural chemicals that exert a hormone-like activity and a variety of biological effects on early development of mouse. In this study, the probable teratogenic effects of RA on CNS have been investigated in pregnant mice ( n = 20) divided into four groups: (1) untreated controls, (2) controls which received a single dose of DMSO, (3) a group that received 40 mg/kg, and (4) a group that received 60 mg/kg of all-trans RA in DMSO, respectively on the eighth day of gestation. Embryos whose dams had received 40 and 60 mg/kg doses of RA, showed malformations and decreased size. At 40 mg/kg dosage level, 50% of the embryos had closed neural tubes while at 60 mg/kg dosage level the neural tube failed to close. The neuroblast mantle layers were disorganized in the 40 mg/kg and even more in the 60 mg/kg exposed group compared to the controls. In mitosis, the density of chromatin was increased in the 60 mg/kg dose group. Compared to controls the 40 and 60 mg/kg dose groups of RA treated dams decreases in the luminal longitudinal and internal measures were observed. Also the thickness of ventricular, mantle and marginal layers was smaller. Wide intercellular spaces due to the degenerated cells at high doses of RA as well as an accumulation of intercellular fluid were observed. Therefore, the wedge shape of neuroepithelium was abolished, preventing the elevation of the neural wall.

Endocrine-Disrupting Chemicals Probed as Potential Pathways to Illness

SAN DIEGO—For the last 60 years, humans have been exposed to an increasing number of synthetic compounds in the environment. In fact, more than 80 000 chemicals are used commercially in the United States, found in products ranging from toys and detergents to pesticides and food packaging. Accumulating evidence that some of these widely used chemicals may have hormonelike effects on the body is heightening concerns about their potential long-term health risks, particularly when developing fetuses and neonates are exposed. At a day-long symposium at the Endocrine Society meeting here in June, leading investigators shared the latest findings on the effects of these synthetic chemicals—as well as of naturally occurring substances such as phytoestrogens—that have been reported to have hormonelike activity and may be associated with an increased susceptibility to disease and dysfunction. The potential danger is greatest for “the fragile fetus” (a term coined more than a decade ago by University of California at Berkeley endocrinologist Howard Bern, PhD) and the neonate, which are particularly sensitive to perturbations by endocrine-disrupting chemicals. But such effects are often not apparent for years. “Exposure to chemicals with endocrine-disrupting activity during critical stages of differentiation may have permanent consequences that may not be expressed or detected until later in life,” said Retha R. Newbold, MS, head of developmental endocrinology studies in the Laboratory of Molecular Toxicology at the National Institute of Environmental Health Sciences. There is now greater recognition, she added, that the developing organism may be especially vulnerable to endocrinedisrupting chemicals because of such factors as deficiencies in DNA repair mechanisms, detoxification enzymes that are not completely functional, and a blood-brain barrier that is still being formed.

Activation of nitric oxide synthesis in human endothelial cells by red clover extracts

The unexpected findings of the Women's Health Initiative trial, where surrogate cardiovascular risk markers have failed to predict the cardiovascular performance of hormone therapy, showing no reduction of cardiovascular disease in postmenopausal women receiving hormonal preparations inducing a favorable lipid profile, raise the interest on how molecules with hormone-like activity used for the treatment of menopausal symptoms act on vascular cells. This is particularly important for estrogen-like compounds such as phytoestrogens, whose mechanisms of action may significantly differ from those of other estrogenic compounds. Because endothelial-derived nitric oxide (NO) is a key regulator of vascular tone and atherogenesis as well as a well-characterized estrogen-regulated molecule, we studied the regulation of NO synthesis in cultured human endothelial cells by phytoestrogens contained in red clover extracts. We show that red clover extracts activate NO synthesis in endothelial cells by recruiting transcriptional pathways but are not capable of inducing rapid NO synthesis through nongenomic mechanisms. During prolonged exposures, red clover extracts enhance the expression as well as the activity of endothelial nitric oxide synthase. These effects are mediated by a recruitment of estrogen receptor-beta. Moreover, we show that red clover-derived isoflavones synergize with 17beta-estradiol in increasing endothelial nitric oxide synthase activity and expression, therefore being devoid of antiestrogenic effects in human endothelial cells. These results help to understand the mechanisms of action of phytoestrogens on the cardiovascular system and have relevant clinical implications.

Assessment of estrogenic activity of flavonoids from Mediterranean plants using an in vitro short-term test

Six isoflavones, daidzein (4',7,-dihydroxyisoflavone), genistein (4',5,7-trihydroxyisoflavone), genistin (genistein 7-O-beta-D-glucopyranoside), isoprunetin (4',7-dihydroxy, 5-metoxyisoflavone), isoprunetin 7-O-beta-D-glucopyranoside, isoprunetin 4',7-di-O-beta-D-glucopyranoside and four flavones, luteolin (3',4',5,7-tetrahydroxyflavone), luteolin 7-O-beta-D-glucopyranoside, luteolin 4'-O-beta-D-glucopyranoside, licoflavone C (4',5,7-trihydroxy,8-isoprenylflavone) were purified from Mediterranean plants (Genista morisii and Genista ephedroides) and their estrogenic activity was assessed by a yeast reporter gene assay (Saccharomyces cerevisiae RMY326 ER-ERE). Licoflavone C showed a powerful estrogenic activity at 10(-7) M (0.0338 microg/ml) and it was 47.45% than 10(-8) M 17beta-estradiol (0.00272 microg/ml). The estrogenicity of this flavone was found to be comparable to the activity showed by genistein at 10(-6) M (0.27 microg/ml). This study points out that a glucose substituent in flavones and isoflavones modulates the hormone-like activity in a different way. Isoflavone aglycones showed a more estrogenic activity than the corresponding glucosides. Conversely, the glucosidation made estrogenic the flavone luteolin and the position of substitution differently influenced the estrogenic activity of compounds.

Mechanisms of plant growth stimulation by humic substances: The role of organo-iron complexes

Stimulatory effects of humic substances (HS) on plant growth have been observed and widely documented. Studies have often shown positive effects on seed germination, root initiation and total plant biomass. The consistency of these observations has been uncertain, predominantly due to the lack of understanding of the plant growth promotion mechanism. Often these effects have been attributed to a direct effect of plant growth hormones; whereas in other instances the term “hormone-like activity” has been used to describe the plant growth stimulation (Chen and Aviad, Humic Substances in Soil and Crop Sciences: Selected Readings, American Society of Agronomy, Soil Science Society of America, 1990; Nardi et al., Humic Substances in Terrestrial Ecosystems, Elsevier Science B.V., 1996). Yet, investigators have been unable to prove that plant growth regulators are present in HS preparations, or the evidence provided remains unconvincing. An alternative hypothesis suggesting that growth enhancement of plants grown in nutrient solution (NS) containing HS is the result of improved micronutrient availability, Fe in particular, has been postulated and tested in the present study. Nutrient solutions containing N, P, K, Ca, Mg, S, B, Mo, Cu, Mn, Zn, and Fe at concentrations considered to be optimal for plant growth were tested for solubility of the Fe, Zn, and Mn, 7 days after preparation. In addition to control solutions at pH values of 5, 6, 7, and 7.5, 0 to 200 mg L−1 of leonardite humic acid (HA) were added to the solutions and they were tested for Fe and Zn solubility. The HA greatly enhanced the maintenance in solution of Fe, in all the tested solutions, and Zn at pH 7.5. Mn mostly remained in solution in its inorganic forms. Plant growth experiments were carried out on both dicotyledonous plants (melons and soybean) and monocotytedonous Poaceae plants (ryegrass), due to the major difference in their Fe uptake mechanism. Plants grown in the absence of Fe exhibited severe Fe deficiency that could only partially be corrected with the addition of mineral Fe salts. The addition of HA or fulvic acid (FA) without addition of Fe, and Zn resulted in partial growth enhancement and correction of Fe deficiency, or none of the two, in the various experiments. This suggests that the growth enhancement effect observed in solutions containing Fe, Zn, and HS was related to the micronutrients rather than to phytohormones. However, the addition of Fe, Zn and either EDTA, HA or FA resulted in healthy, chlorophyll rich plants and enhanced growth, thereby providing evidence that improved Fe, and possibly Zn nutrition is a major mechanism of plant growth stimulation by HS. The use of the term hormone-like activity could be the result of the similarity of the physiological effects obtained in plants enjoying sufficient supply of Fe and Zn.

Bisphenol A affects glucose transport in mouse 3T3‐F442A adipocytes

Recently, environmental chemicals have appeared in daily human life, and these chemicals have been incidentally taken in by humans. The serum concentrations of some of these chemicals have been found to be associated with the onset and incidence rate of diabetes mellitus. It has been suggested that one of the environmental chemicals, bisphenol A (BPA), has hormone-like activity. It has also been demonstrated that some hormones affect insulin resistance and fat distribution in the body. To study the effects of these environmental chemicals on glucose metabolism, the effect of BPA on glucose transport in mouse 3T3-F442A adipocytes was investigated. The 3T3-F442A adipocytes were incubated with various concentrations of BPA in a medium. Deoxyglucose uptake assay was performed with and without insulin. Immunoblot analysis was performed with a glucose transporter (GLUT) 4-specific antibody and antiphosphotyrosine antibody. The BPA treatment enhanced basal and insulin-stimulated glucose uptake, and caused an increased amount of GLUT4 protein. Thus, the enhanced glucose uptake resulting from the BPA treatment was at least partially due to the increased amount of GLUT4. Tyrosine phosphorylation of insulin receptor substrate-1 with insulin stimulation was not significantly affected. In conclusion, it was demonstrated that BPA, one of the chemicals that we intake incidentally, affects the glucose transport in adipocytes, and also that the environmental chemicals may be identified as one of the environmental factors that affect diabetes and obesity.

Biological activities of tethered equine chorionic gonadotropin (eCG) and its deglycosylated mutants.

Equine chorionic gonadotropin (eCG), which consists of highly glycosylated alpha- and beta-subunits, is a unique member of the gonadotropin family because it elicits response characteristics of both follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in species other than the horse. In this study, recombinant tethered-eCG as well as its deglycosylated mutants were produced to determine if alpha- and beta- subunits can be synthesized as a single polypeptide chain (tethered-eCG) and display biological activity. We found that tethered-eCG (T- betaalpha) had both LH- and FSH-like activities comparable to dimeric eCG. Luteinizing hormone-like activity of tethered-eCGs deglycosylated at Asn(56) (T-betaalpha56) was decreased. In contrast, LH-like activity of eCG without O-glycosylated carboxyl-terminal peptide (CTP) (T-betacalpha) was slightly decreased but still similar to T-betaalpha. Double mutation at Asn(56) and CTP (T-betacalpha56) caused marked decrease in the activity, indicating that both glycosylations at Asn(56) and CTP are involved in LH-like activity in the tethered form. Interestingly, FSH-like activity remained in all deglycosylated eCG mutants (T-betaalpha56, T-betacalpha and T-betacalpha56) as well as T-betaalpha. The biological roles of oligosaccharides at Asn(56) of eCG alpha-subunit and O-linked peptide of beta-subunit appear to be different in LH- and FSH-like activities in tethered-eCG.

Differential regulation of EIN3 stability by glucose and ethylene signalling in plants.

Glucose is a global regulator of growth and metabolism that is evolutionarily conserved from unicellular microorganisms to multicellular animals and plants. In photosynthetic plants, glucose shows hormone-like activities and modulates many essential processes, including embryogenesis, germination, seedling development, vegetative growth, reproduction and senescence. Genetic and phenotypic analyses of Arabidopsis mutants with glucose-insensitive (gin) and glucose-oversensitive (glo) phenotypes have identified an unexpected antagonistic interaction between glucose and the plant stress hormone ethylene. The ethylene-insensitive etr1 and ein2 mutants have glo phenotypes, whereas the constitutive ethylene signalling mutant ctr1 is allelic to gin4 (refs 4, 5). The precise molecular mechanisms underlying the complex signalling network that governs plant growth and development in response to nutrients and plant hormones are mostly unknown. Here we show that glucose enhances the degradation of ETHYLENE-INSENSITIVE3 (EIN3), a key transcriptional regulator in ethylene signalling, through the plant glucose sensor hexokinase. Ethylene, by contrast, enhances the stability of EIN3. The ein3 mutant has a glo phenotype, and overexpression of EIN3 in transgenic Arabidopsis decreases glucose sensitivity.

Acutely elevated vasopressin increases circulating concentrations of cortisol and aldosterone in fasting northern elephant seal (Mirounga angustirostris) pups

The physiological actions of vasopressin (VP) in marine mammals are not well defined. To help elucidate its hormonal and renal effects in this group of mammals, northern elephant seal (Mirounga angustirostris) pups (N=7; 99+/-4 kg) were first infused with 0.9% saline (control; 220 ml), followed 24 h later with VP (as a 20 ng kg(-1) bolus, then 2 ng kg(-1) min(-1) for approximately 35 min in 225+/-16 ml saline). During both control and VP periods, blood samples were collected prior to infusion, and 15, 30, 60, 120 min and 24 h after infusion to examine the hormonal responses of the pups to VP. Renal responses were quantified from 24 h urine samples obtained prior to infusion (control) and 24 h post-infusion. Compared to the control period, infusion of VP increased plasma concentrations of cortisol over a 120 min period and aldosterone over 30 min, while plasma renin activity (PRA) was decreased for a 120 min period. The plasma urea:creatinine ratio was elevated following infusion of VP. Urine output and osmotic clearance were increased by 69+/-18% (mean +/- S.E.M.) and 36+/-10%, respectively, but free water clearance and glomerular filtration rate were not significantly altered 24 h post-infusion of VP. Solute (osmolality, Na(+), K(+) and Cl(-)) excretion and fractional excretion of electrolytes were also increased when compared to control values. The increase in cortisol concentration suggests that VP may possess corticotropin releasing hormone-like activity in elephant seals. If osmotic diuresis and natriuresis are typical consequences of elevated [VP] in fasting pups, then not increasing VP normally during the fast may serve as a protective mechanism to avoid the potential loss of Na(+) induced by elevated [VP]. Therefore, under natural fasting conditions, pups may be highly sensitive to small changes in [VP], resulting in the maintenance of water and electrolyte balance.

Low-molecular-weight organic acids and hormone-like activity of dissolved organic matter in two forest soils in N Italy.

Concentrations of aliphatic acids, phenolic acid, and inorganic nutrients, as well as hormone-like activity, were determined in soil dissolved organic matter (DOM) from two forest sites in N Italy showing differing degrees of silver fir regeneration. In the site where silver fir recruitment was abundant, humification processes prevailed, and the soil DOM had a high content in aliphatic and phenolic acids. This enhanced the hormone-like activity in the soil, which could in turn promote growth of silver fir seedlings. In the site with poor fir recruitment, the soil DOM underwent rapid mineralization and was richer in inorganic nutrients, but had lower concentrations of aliphatic and phenolic acids, and lower hormone-like activity.

Health Effect of Polychlorinated Biphenyls and Related Compounds

Polychlorinated biphenyls (PCBs) and dioxins have polluted the environment for about 50 years and parts of them have transferred to human being through food chain. Humans are already contaminated with chlorinated compounds at relatively high levels. Yusho PCB poisoning occurred at Northern Kyushu in 1968 and the patients have been suffering from various symptoms for 35 years. High concentrations of toxic PCBs and polychlorinated dibenzofurans in the patients have been very gradually decreased to the levels of only 2 to 6 times higher than those of normal persons. Typical Yusho symptoms of acneiform eruption, dermal pigmentation and increased eye discharge were very gradually recovered with lapse of several years. However, enzyme and/or hormone-mediated signs of high serum triglyceride, high serum thyroxin, immunoglobulin disorder and others are persistently maintained for more than 30 years. Recent tolerable daily intake of dioxins were determined from their hormone-like activities, such as decreased sperm count, immune suppression, increased genital malformation and neurobehavioral effects in the offspring of animals. PCBs are important factors on considering dioxin toxicities to human for the reason that half the dioxin toxicity in normal persons consists of the toxicity of coplanar PCBs. Recent epidemiological studies have reported that higher levels of PCBs and related compounds in the human body associated with various health effects, such as lowering intelligence quotient levels, disorder of thyroid gland, higher rate of endometriosis in women, declining thyroid hormone levels, higher rate of diabetes in pregnant women, lowering age at menarche, and altering play behavior in children at school age.

Synthesis of a Castasterone/Ponasterone Hybrid Compound and Evaluation of Its Molting Hormone-Like Activity

Castasterone/ponasterone hybrid compound, (20R)-2α,3α,20, 22-tetrahydroxy-5α-cholestan-6-one, was newly synthesized from the corresponding α-alkoxystannane, in which the α-hydroxy group was protected with a benzyloxymethyl ether, and 6,6-ethylenedioxy-2α,3α-isopropylidenedioxy-5α-pregnan-20-one. The inhibition of the incorporation of [3H]ponasterone A into two insect cell lines was examined, and the concentrations of this hybrid compound required to give 50% inhibition were determined to be 0.29μM and 0.89μM for Kc and Sf-9 cells, respectively. This hybrid compound was 10 times more potent than ecdysone, but 100-200 times less potent than ponasterone A. The potency of this compound was equivalent to inokosterone against Sf-9 cells. The molting hormonal effect of this compound was also evaluated in the cultured integument system of the rice stem borer Chilo suppressalis for the induction of chitin synthesis, and the 50% effective concentration was determined to be about 10μM, equipotent to ecdysone.

Hormone-like activities of humic substances in different forest ecosystems.

• In order to understand the phytohormone-like activity of humic substances (HS) with respect to vegetation, different forest ecosystems were considered. For this study, 32 soil horizons from a large area of northern Italy typical for silver fir ( Abies alba ) were evaluated. • The development of organic and humic matter in the five silver fir forest types was assessed by quantifying chemical and biochemical parameters. Phytohormone-like activity of HS was evaluated by measuring auxin-like (indoleacetic acid (IAA)-like) and gibberellin-like (GA-like) activities as well as invertase and peroxidase activities. • Differences between ecosystems were obtained by comparing the five silver fir types with previous work in beech forests, which investigated three large groups: thermophilous, mesophilous and acidophilous. In these eight forests it was reconfirmed that acid conditions were essential for the release of the IAA-like activity, whereas neutral conditions promoted GA-like activity. • Humic substances are of ecological importance because their phytohormone-like activity stands out not only within the five silver fir forest types, but also between the silver fir and beech types. Our results demonstrated a different collocation of the hormone-like activity along the profile in accordance with the different growth conditions, seed germination and the first stage of seedling growth. Humic substances evoke the greatest response on seed germination and on the first stage of seedling growth, particularly in stressed environmental conditions.