Changes in backscattered ultrasonic envelope statistics as a function of thrombus age: an in vitro study.

It is necessary to determine the age of thrombi in planning clinical treatment for thrombolysis. Ultrasound imaging can potentially be used to evaluate thrombus age in real time. The backscattered signals from thrombi may contain useful information regarding their age. On the basis of the randomness of ultrasound backscattering, this study explored changes in backscattered US statistics as a function of thrombus age. Porcine blood samples were used for the in vitro induction of fresh thrombi (day 0) with hematocrits ranging from 0%-40% and aged thrombi (days 0-8) with a hematocrit of 40%. Each thrombus was imaged using a pulse-echo ultrasound scanner equipped with a 7.5-MHz linear array transducer to acquire raw backscattered signals for B-mode and Nakagami imaging, by which the backscattered statistics were visualized. Hematoxylin and eosin staining and scanning electron microscopy were used to observe the histology of fresh and aged thrombi. The results indicated that a decrease in the number of red blood cells in the thrombus caused by the aging effect was observed in the in vitro model, indicating that the proposed model could simulate the structural changes in the thrombus during aging. Compared with fresh thrombi with various hematocrits, the aged thrombi exhibited a trend toward more substantial decreases in the Nakagami parameter with increasing thrombus age (the Nakagami parameter decreased from 1.1 to 0.6 as thrombus age increased from day 0 to day 8), indicating that thrombus aging causes the backscattered statistics to follow a pre-Rayleigh distribution to a high degree. This finding may be applied to the determination of thrombus age using conventional ultrasound imaging in the future.

Multi-criteria analysis via the VIKOR method for prioritizing land-use restraint strategies in the Tseng-Wen reservoir watershed.

It is important to adopt proper water and soil conservation and land-use planning in a watershed for lowering adverse impacts on reservoir water quality. Although reservoir watersheds occupy a large amount of land in Taiwan, high population density has exerted development pressures on such land. Therefore, the priority ranking of land-use restrictions for the subdivisions with different degrees of environmental vulnerability is necessary in watershed management. Since there are several criteria for evaluating the potential environmental impact from the subdivisions, multi-criteria analysis was applied as a technique for solving these problems in this study. The VIKOR method was applied to determine the best feasible solution according to the selected criteria, including geographical and meteorological factors. The objective of this study was to establish the priority ranking of land-use restrictions in the Tseng-Wen reservoir wastershed in southern Taiwan. The results show that subdivisions close to the outlet or reservoir area should have the priority of land-use restrictions.

The orphan nuclear receptor chicken ovalbumin upstream promoter-transcription factor II is a critical regulator of adipogenesis.

The orphan nuclear receptor chicken ovalbumin upstream promoter-transcription factor II (COUP-TFII; Nr2f2) is expressed in adipose tissue in vivo and declines during differentiation. Overexpression of COUP-TFII prevents adipogenesis, whereas shRNA-mediated reduction of COUP-TFII promotes differentiation, as shown by increased lipid accumulation and elevated expression of fat cell marker proteins. Furthermore, reduction of COUP-TFII allows uncommitted fibroblasts to be differentiated into fat cells. COUP-TFII represses the expression of a number of proadipogenic factors in adipocytes, with direct action noted at the CAAT enhancer-binding protein alpha promoter. We show that COUP-TFII acts downstream of hedgehog signaling and is required for the full antiadipogenic effect of this pathway. This effect is mediated in part by interaction with GATA factors. COUP-TFII and GATA2 are physically associated and repress target gene expression in an additive manner. Taken together, our data demonstrate that COUP-TFII represents an endogenous suppressor of adipogenesis, linking antiadipogenic extracellular signals to the core transcriptional cascade.

Interferon regulatory factors are transcriptional regulators of adipogenesis.

We have sought to identify transcriptional pathways in adipogenesis using an integrated experimental and computational approach. Here, we employ high-throughput DNase hypersensitivity analysis to find regions of altered chromatin structure surrounding key adipocyte genes. Regions that display differentiation-dependent changes in hypersensitivity were used to predict binding sites for proteins involved in adipogenesis. A high-scoring example was a binding motif for interferon regulatory factor (IRF) family members. Expression of all nine mammalian IRF mRNAs is regulated during adipogenesis, and several bind to the identified motifs in a differentiation-dependent manner. Furthermore, several IRF proteins repress differentiation. This analysis suggests an important role for IRF proteins in adipocyte biology and demonstrates the utility of this approach in identifying cis- and trans-acting factors not previously suspected to participate in adipogenesis.

The adipokine lipocalin 2 is regulated by obesity and promotes insulin resistance.

OBJECTIVE: We identified lipocalin 2 (Lcn2) as a gene induced by dexamethasone and tumor necrosis factor-alpha in cultured adipocytes. The purpose of this study was to determine how expression of Lcn2 is regulated in fat cells and to ascertain whether Lcn2 could be involved in metabolic dysregulation associated with obesity. RESEARCH DESIGN AND METHODS: We examined Lcn2 expression in murine tissues and in 3T3-L1 adipocytes in the presence and absence of various stimuli. We used quantitative Western blotting to observe Lcn2 serum levels in lean and obese mouse models. To assess effects on insulin action, we used retroviral delivery of short hairpin RNA to reduce Lcn2 levels in 3T3-L1 adipocytes. RESULTS: Lcn2 is highly expressed by fat cells in vivo and in vitro. Expression of Lcn2 is elevated by agents that promote insulin resistance and is reduced by thiazolidinediones. The expression of Lcn2 is induced during 3T3-L1 adipogenesis in a CCAAT/enhancer-binding protein-dependent manner. Lcn2 serum levels are elevated in multiple rodent models of obesity, and forced reduction of Lcn2 in 3T3-L1 adipocytes improves insulin action. Exogenous Lcn2 promotes insulin resistance in cultured hepatocytes. CONCLUSIONS: Lcn2 is an adipokine with potential importance in insulin resistance associated with obesity.

Targeted elimination of peroxisome proliferator-activated receptor gamma in beta cells leads to abnormalities in islet mass without compromising glucose homeostasis.

The nuclear hormone receptor peroxisome proliferator-activated receptor gamma (PPAR gamma) is an important regulator of lipid and glucose homeostasis and cellular differentiation. Studies of many cell types in vitro and in vivo have demonstrated that activation of PPAR gamma can reduce cellular proliferation. We show here that activation of PPAR gamma is sufficient to reduce the proliferation of cultured insulinoma cell lines. We created a model with mice in which the expression of the PPARG gene in beta cells was eliminated (beta gamma KO mice), and these mice were found to have significant islet hyperplasia on a chow diet. Interestingly, the normal expansion of beta-cell mass that occurs in control mice in response to high-fat feeding is markedly blunted in these animals. Despite this alteration in beta-cell mass, no effect on glucose homeostasis in beta gamma KO mice was noted. Additionally, while thiazolidinediones enhanced insulin secretion from cultured wild-type islets, administration of rosiglitazone to insulin-resistant control and beta gamma KO mice revealed that PPAR gamma in beta cells is not required for the antidiabetic actions of these compounds. These data demonstrate a critical physiological role for PPAR gamma function in beta-cell proliferation and also indicate that the mechanisms controlling beta-cell hyperplasia in obesity are different from those that regulate baseline cell mass in the islet.

C/EBPalpha induces adipogenesis through PPARgamma: a unified pathway.

PPARgamma and C/EBPalpha are critical transcription factors in adipogenesis, but the precise role of these proteins has been difficult to ascertain because they positively regulate each other's expression. Questions remain about whether these factors operate independently in separate, parallel pathways of differentiation, or whether a single pathway exists. PPARgamma can promote adipogenesis in C/EBPalpha-deficient cells, but the converse has not been tested. We have created an immortalized line of fibroblasts lacking PPARgamma, which we use to show that C/EBPalpha has no ability to promote adipogenesis in the absence of PPARgamma. These results indicate that C/EBPalpha and PPARgamma participate in a single pathway of fat cell development with PPARgamma being the proximal effector of adipogenesis.