Regulation of LKB1 expression by sex hormones in adipocytes.

OBJECTIVE: In the adipose tissue, activation of AMP-activated protein kinase (AMPK) by phosphorylation favours local fatty acid oxidation and inhibition of lipogenesis. We have previously shown that the potent androgen dihydrotestosterone (DHT) can inhibit phosphorylation of AMPK in adipose tissue and 3T3-L1 adipocytes in a dose-dependent manner. This negative effect of DHT was reversed by oestrogen treatment. The purpose of this current study was to determine the underlying mechanisms whereby androgens and oestrogens can regulate AMPK phosphorylation in adipocytes, and whether this mechanism is receptor dependent. RESULTS: Phosphorylation of AMPK was assessed by western blot in cells treated for 24 h with testosterone or DHT (1-1000 nM). Testosterone and DHT significantly inhibited basal phosphorylation of AMPK. Addition of the androgen receptor antagonist Flutamide (1 µM) to the media reversed the negative effect of testosterone and DHT by returning AMPK phosphorylation levels to those of basal. To further dissect the mechanism underlying AMPK inhibition by testosterone or DHT, we examined the mRNA expression of the upstream activator of AMPK, namely LKB1. Testosterone and DHT treatment of murine 3T3-L1 or human SGBS adipocytes for 24 h significantly decreased the mRNA expression of LKB1. In contrast, 17ß-estradiol treatment increased LKB1 mRNA, an effect mediated by oestrogen receptor alpha. CONCLUSION: We conclude that regulation of AMPK phosphorylation by androgens and oestrogens is receptor-dependent, and demonstrate for the first time that LKB1 is regulated by sex hormones in adipocytes.

Estrogen and adiposity--utilizing models of aromatase deficiency to explore the relationship.

Estrogen has an important role to play in energy homeostasis in both men and mice. Lack of estrogen results in the development of a metabolic syndrome in humans and rodents, including excess adiposity, hepatic steatosis (in male but not female aromatase knockout (ArKO) mice) and insulin resistance. Estrogen replacement results in a prompt reversal of the energy imbalance symptoms associated with estrogen deficiency. A corollary to the perturbed energy balance observed in the ArKO mouse is the death by apoptosis of dopaminergic neurons in the hypothalamic arcuate nucleus of male ArKO mice, an area of the brain pivotal to the regulation of energy uptake, storage, and mobilisation. An extension of our work exploring the relationship between estrogen and adiposity has been to examine the role played by androgens in energy balance. We have demonstrated that an increased androgen to estrogen ratio can promote visceral fat accumulation in the rodent by inhibiting AMPK activation and stimulating lipogenesis. Therefore, understanding the regulation of energy homeostasis is becoming an increasingly fascinating challenge, as the number of contributors, their communications, and the complexity of their interactions, involved in the preservation of this equilibrium continues to increase. Models of aromatase deficiency, both naturally occurring and engineered, will continue to provide valuable insights into energy homeostasis.

Influence of salinity on bioremediation of oil in soil.

Spills from oil production and processing result in soils being contaminated with oil and salt. The effect of NaCl on degradation of oil in a sandy-clay loam and a clay loam soil was determined. Soils were treated with 50 g kg(-1) non-detergent motor oil (30 SAE). Salt treatments included NaCl amendments to adjust the soil solution electrical conductivities to 40, 120, and 200 dS m(-1). Soils were amended with nutrients and incubated at 25 degrees C. Oil degradation was estimated from the quantities of CO(2) evolved and from gravimetric determinations of remaining oil. Salt concentrations of 200 dS m(-1) in oil amended soils resulted in a decrease in oil mineralized by 44% for a clay loam and 20% for a sandy-clay loam soil. A salt concentration of 40 dS m(-1) reduced oil mineralization by about 10% in both soils. Oil mineralized in the oil amended clay-loam soil was 2-3 times greater than for comparable treatments of the sandy-clay loam soil. Amending the sandy-clay loam soil with 5% by weight of the clay-loam soil enhanced oil mineralization by 40%. Removal of salts from oil and salt contaminated soils before undertaking bioremediation may reduce the time required for bioremediation.

Adsorption and desorption of metolachlor and metolachlor metabolites in vegetated filter strip and cultivated soil.

Previous studies have indicated that dissolved-phase metolachlor [2-chloro-N-(2-ethyl-6-methylphenyl)-N-(methoxy-1-methylethyl) acetamide] transported in surface runoff is retained by vegetative filter strips to a greater degree than either metolachlor oxanilic acid 12-[(2-ethyl-6-methylphenyl) (2-methoxy-1-methylethyl)amino]-2-oxo-acetic acid] (OA) or metolachlor ethanesulfonic acid [2-[(2-ethyl-6-methylphenyl) (2-methoxy-1-methylethyl-1)amino]-2-oxoethanesul-fonic acid] (ESA), two primary metabolites of metolachlor. Adsorption-desorption of ESA and OA in vegetated filter strip soil (VFSS) has not been evaluated, yet these data are required to assess the mobility of these compounds in VFSS. The objective of this experiment was to compare metolachlor, ESA, and OA adsorption and desorption parameters between VFSS and cultivated soil (CS). Adsorption and desorption isotherms were determined using the batch equilibrium procedure. With the exception of a 1.7-fold increase in organic carbon content in the VFSS, the evaluated chemical and physical properties of the soils were similar. Sorption coefficients for metolachlor were 88% higher in VFSS than in CS. In contrast, sorption coefficients for ESA and OA were not different between soils. Relative to metolachlor, sorption coefficients for ESA and OA were at least 79% lower in both soils. Metolachlor desorption coefficients were 59% higher in the VFSS than in the CS. Desorption coefficients for ESA and OA were not different between soils. Relative to metolachlor, desorption coefficients for ESA and OA were at least 66% lower in both soils. These data indicate that the mobility of ESA and OA will be greater than metolachlor in both soils. However, higher organic carbon content in VFSS relative to CS may limit the subsequent transport of metolachlor from the vegetated filter strip.

Water flow patterns in subsurface flow constructed wetlands designed for on-site domestic wastewater treatment.

Bypass flow in subsurface flow constructed wetlands, which may be related to several wetland characteristics, reduces detention time and may result in inadequately treated wastewater. Subsurface flow constructed wetlands, 2.3 m wide by 4.8 m long and containing a gravel matrix, were used to investigate the impact of wetland depth, inlet pipe location, loading volume, and plants on water flow. Flow patterns were determined using blue dye or bromide as tracers. The blue dye adsorbed to the gravel and was not an effective tracer for following water movement. Water dispersed as it flowed through the wetland, and approximately two pore volumes of added water were required to displace approximately 99% of the bromide tracer added as a pulse. In 17 and 25 cm deep wetlands, water flow was uniform with depth, and inlet depth had little influence on water flow patterns. Water flow in a 40 cm deep wetland was not uniform with depth for either inlet pipe placement. The presence of plants caused preferential water flow around root masses, thereby, limiting their potential to interface with wastewater. Water mixing by dispersion from the surface to deeper depths may enhance aeration. Bromide was first detected in effluent after only 0.5 pore volumes of tap water had been added. This indicates that detention time for some wastewater would be less than expected, since plug flow is usually assumed in subsurface flow constructed wetlands. This occurrence should be considered for time-dependent treatments, such as fecal coliform and biological oxygen demand reduction.

Surfacing of domestic wastewater applied to soil through drip tubing and reduction in numbers of Escherichia coli.

Drip tubing is a technology that is increasing in use. The effectiveness of such systems in distributing the wastewater uniformly through the soil matrix, providing adequate removal of bacteria from wastewater, and keeping wastewater from reaching the soil surface has not been adequately evaluated. Experiments were conducted at two sites that had used drip tubing for approximately 3 years. This 3-year-old drip tubing and newly installed tubing were used in this investigation. A solution containing Brilliant Blue FCF dye and Escherichia coli, at an approximate concentration of 1 x 10(6) cells ml(-1), was applied to the sites through drip emitters. Reduction i n Escherichia coli populations reaching the soil surface was generally less than 10%. The route of travel for the solution reaching the soil surface was consistently along preferential flow paths and not uniformly through the soil matrix. Instances of water reaching the soil surface for drip tubing installed at 15 cm was nearly 50%. Increasing burial of the drip tubing from 15 to 30 cm nearly eliminated water surfacing. A 31 per emitter dose of water, immediately following drip line installation later increased instances of water reaching the soil surface for drip tubing buried at 30 cm. The volume of water applied per dose had little effect on the number of times water reached the soil surface. Inherent soil structural characteristics limited the drip tubing's ability to uniformly distribute water and adsorb bacteria. Drip tubing installation to 30 cm may be an important practice to reduce public health hazards from the likelihood of wastewater surfacing.

Endometrial Intracrinology--generation of an estrogen-dominated microenvironment in the secretory phase of women.

CONTEXT: The human endometrium is a complex multicellular tissue subject to cyclical fluctuations in ovarian-derived steroid hormones. Fertile cycles are characterized by differentiation (decidualization) of endometrial stromal cells (ESC). OBJECTIVE: To determine the impact of human stromal cell decidualization on biosynthesis and secretion of estrogens. DESIGN: Primary cell culture was used. Cells were decidualized in vitro. Some cultures were treated with an aromatase inhibitor. SETTING: A University Research Institute. PATIENTS: Primary ESC were derived from women with normal menstrual cycles (n = 12). None of the women were receiving hormonal therapy or suffering from endometriosis. MAIN OUTCOME MEASUREMENTS: Expression of mRNA and protein encoded by the aromatase (CYP19A1) and 3-ß-hydroxysteroid dehydrogenase (HSD3B1) genes was assessed. Aromatase activity was measured using a tritiated water assay; steroid metabolism was determined using thin layer chromatography. Estrone (E1) and estradiol (E2) in cell culture media were measured by ELISA. RESULTS: Decidualization induced a two-fold increase in aromatase mRNA. Aromatase protein was only detected in decidualized ESC. 3-ß-Hydroxysteroid dehydrogenase protein was present in ESC both before and after decidualization; concentrations appeared unchanged. The existence of functional aromatase in decidualized ESC was confirmed; E1 and E2 were secreted into culture media in decidualized ESC and concentrations were reduced when cells were incubated with an aromatase inhibitor. Decidualization resulted in reduced metabolism of E2 and an increase in the ratio of E2:E1. CONCLUSIONS: Decidualization is characterized by an increase in aromatase expression/activity favoring the generation of an E2-dominated estrogen microenvironment within the endometrial stroma.