Angiotensin II-responsive element is the insulin-responsive element in the adipocyte fatty acid synthase gene: role of adipocyte determination and differentiation factor 1/sterol-regulatory-element-binding protein 1c.

We have previously shown that angiotensin II (Ang II) increases the expression of the gene encoding adipocyte fatty acid synthase (FAS). Here we investigate the mechanism responsible for increased FAS gene transcription by Ang II. We demonstrate that Ang II increased luciferase activity by 3-fold in 3T3-L1 adipocytes transfected with fusion constructs linking the FAS promoter to the luciferase reporter gene. Interestingly, we mapped the Ang II regulatory sequences to the insulin-responsive region (E box) in the proximal FAS promoter. The E box alone was able to mediate Ang II responsiveness when linked to a heterologous promoter. However, this response was lost when mutations that abolished the binding of the E box to its transcription factors were introduced. Using adenoviral overexpression of a dominant-negative form of adipocyte determination and differentiation factor 1 (ADD1), a transcription factor that binds to the insulin-responsive E box, we demonstrated that ADD1 was required for Ang II regulation of the FAS gene in 3T3-L1 adipocytes. Furthermore, ADD1 expression was also up-regulated by Ang II. With the use of transfections as well as glucose transport assays, we further demonstrated that Ang II stimulation of the FAS gene was dependent on glucose. In conclusion, this is the first report that Ang II regulates adipocyte FAS gene transcription via insulin response sequences in a glucose-dependent manner and that this regulation is mediated at least in part via the ADD1 transcription factor.

Healing bittersweet legacies: revisiting contextual family therapy for grandparents raising grandchildren in crisis.

Over the past decade, grandparents have become primary caregivers for their grandchildren at an unprecedented rate. Drug/alcohol abuse, poor mental health, financial instability, chronic illness, and early death have left parents unavailable for childrearing, leaving their elder generation responsible for tending vulnerable children. While grandparents' resources have been overtaxed with this arrangement, family therapists have been caught off guard in adapting traditional family therapy methods to this population. This paper revisits Contextual Family Therapy for what it has to offer grandparents in crisis. The purpose is to restore appropriate "give and take" while compensating for a "skipped" parenting generation.

Diazoxide down-regulates leptin and lipid metabolizing enzymes in adipose tissue of Zucker rats.

We have previously reported that attenuation of hyperinsulinemia by diazoxide (DZ), an inhibitor of glucose-mediated insulin secretion, increased insulin sensitivity and reduced body weight in obese Zucker rats. These findings prompted us to investigate the effects of DZ on key insulin-sensitive enzymes regulating adipose tissue metabolism, fatty acid synthase (FAS), and lipoprotein lipase (LPL), as well as on circulating levels of leptin. We also determined the direct effects of diazoxide on FAS in 3T3-L1 adipocytes. Seven-week-old female obese and lean Zucker rats were treated with DZ (150 mg/kg/d) or vehicle (C, control) for a period of 6 wk. Changes in plasma parameters by DZ include significant decreases in triglycerides, free fatty acids, glucose, and insulin, consistent with our previous reports. DZ obese rats exhibited lower plasma leptin levels (P<0.03) compared to their C animals. DZ significantly reduced adipose tissue FAS activity in both lean (P<0.0001) and obese (P<0.01) animals. LPL mRNA content was also decreased significantly in DZ-treated obese animals (P<0.009) as compared to their respective controls without a significant effect on lean animals. The possibility that DZ exerted a direct effect on adipocytes was further tested in cultured 3T3-L1 adipocytes. Although diazoxide (5 microM) alone did not change FAS activity in cultured 3T3-L1 adipocytes, it significantly attenuated insulin's effect on FAS activity (P<0.001). We demonstrate that DZ regulates key insulin-sensitive enzymes involved in regulation of adipose tissue metabolism. These findings suggest that modification of insulin-sensitive pathways can be therapeutically beneficial in obesity management.

Glucose induces expression of stearoyl-CoA desaturase in 3T3-L1 adipocytes.

Stearoyl-CoA desaturase (SCD; EC 1.14.99.5) is a key enzyme in the synthesis polyunsaturated fatty acids. Liver and ose tissue are the predominant sites of SCD expression. Regulation of tic SCD by various nutritional and hormonal ors, such as insulin, dietary carbohydrates and polyunsaturated fatty s, has been well documented. Little is known, ver, about adipocyte SCD regulation despite high levels of SCD activity adipose tissue. The present study was gned to investigate SCD regulation in adipocytes by examining the cts of glucose and insulin on SCD expression. We rt here that glucose availability directly increased SCD gene scription in 3T3-L1 adipocytes. This response was pendent of insulin, and insulin alone in the absence of glucose had no ct on SCD mRNA levels. SCD thus represents a l model in which to investigate the mechanisms of direct regulation of expression by glucose in adipose cells.

Insulin increases fatty acid synthase gene transcription in human adipocytes.

The purpose of this study was to investigate the molecular mechanism whereby insulin increases expression of a key de novo lipogenic gene, fatty acid synthase (FAS), in cultured human adipocytes and hepatoma cells. RNA isolated from cultured adipocytes or from Hep G2 cells treated with or without insulin (20 nM) was analyzed. In addition, run-on transcription assays and measurements of RNA half-life were performed to determine the controlled step in FAS gene regulation by insulin. We demonstrated that FAS mRNA was expressed in both Hep G2 cells and human adipocytes. Insulin induced an approximately five- and three-fold increase in FAS mRNA content in adipocytes and hepatoma cells, respectively. Similar regulation of FAS was observed in adipocytes from lean and obese human subjects. Furthermore, we demonstrated that the induction of human FAS expression by insulin was due to increased transcription rate of the FAS gene in human adipocytes, whereas mRNA stabilization accounted for increased FAS mRNA content in hepatoma cells. In conclusion, we report here for the first time expression of human FAS mRNA and its specific transcriptional induction by insulin in cultured human adipocytes.

Angiotensinogen gene expression in adipose tissue: analysis of obese models and hormonal and nutritional control.

Synthesis of angiotensin II (ANG II) has recently been described in adipose cells and has been linked to regulation of adiposity. Angiotensinogen (AGT), the substrate from which ANG II is formed, was previously shown to be elevated in adipose tissue of obese (ob/ob and db/db) mice and regulated by nutritional manipulation. It is unknown, however, whether overexpression of adipose AGT can be extended to other models of obesity and whether hormonal and/or nutritional factors directly regulate AGT expression in adipocytes. We investigated these possibilities by analyzing AGT mRNA levels in adipose tissue of obese Zucker rats, viable yellow (Avy) mice, and humans and by treating 3T3-L1 adipocytes with insulin, glucose, and a beta-adrenergic agonist. We demonstrate that AGT mRNA is decreased by approximately 50 and 80%, respectively, in adipose tissue of obese vs. lean Zucker rats and Avy mice. We also report that AGT is expressed at variable levels in human adipose tissue. Finally, we show that AGT mRNA is upregulated by insulin and downregulated by beta-adrenergic stimulation in adipocytes.

Angiotensin II increases lipogenesis in 3T3-L1 and human adipose cells.

Angiotensin II (Ang II) is one of numerous hormones recently shown to be synthesized and secreted by adipose cells. Although the function of Ang II in adipose tissue is unknown, several studies indirectly suggest that it may be involved in control of adiposity. Little is known, however, about direct actions of Ang II in adipose cells. To further investigate this issue, we first characterized the type of Ang II receptors in 3T3-L1 adipocytes. We then tested the hypothesis that Ang II exerted direct actions on adipocyte metabolism using both 3T3-L1 and human adipocyte models. We report here that Ang II significantly increased triglyceride content and the activities of two key lipogenic enzymes (fatty acid synthase, FAS and glycerol-3-phosphate dehydrogenase, GPDH) in 3T3-L1 adipocytes, and that these effects were mediated through the type-2 Ang II receptor. We also report that Ang II exerted similar effects in human adipose cells maintained in primary culture. Finally, we demonstrate that Ang II increased the transcription rate of the FAS and ob genes in 3T3-L1 and human adipose cells. These results indicate that Ang II may be involved in control of adiposity through regulation of lipid synthesis and storage in adipocytes.

Combining total quality management and simulation with application to family therapy process design.

This case study shows how a new combined total quality management and simulation procedure was used to design, test, and implement a unique clinical process for brief systems family therapy. A process design procedure was developed that combines total quality management and simulation strategies and tactics. The six strategic check-points of the new procedure emphasize the use of both designed experimentation and process modeling and simulation to find process faults, evaluate process improvement alternatives, and test selected process improvements. Other information needed for process design was gathered from families in therapy, the family therapy literature and observations of student and experienced therapists. Simulation helped identify and compare alternative sequences of therapy process steps. Designed experimentation revealed improvements resulting from the new Brief Systems Family Therapy process as used by doctoral-level family therapy practicum students in a university clinic.

A paradigm for construction of family therapy tasks.

Typically, family therapy task interventions involve direct assignment of specific behaviors, which may or may not encourage client compliance. To enhance the clinician's probability of bringing about progress in treatment, this article broadens the parameters of task intervention to include (a) Direct versus Indirect, (b) Behavioral versus Nonbehavioral, and (c) Paradoxical versus Nonparadoxical dimension alternatives. The resulting 2 x 2 x 2 model, the Family Therapy Task Construction Paradigm, yields eight possible choices for end-of-the-session punctuation. Each option is linked to corresponding treatment contexts and illustrated with examples from productive case vignettes.