Abnormalities in steroid hormones are responsible for the development and prevention of endocrine diseases. Due to their biochemical roles in endocrine system, the quantitative evaluation of steroid hormones is needed to elucidate altered expression of steroids. Gas chromatographic-mass spectrometric (GC-MS) profiling of 70 urinary steroids, containing 22 androgens, 18 estrogens, 15 corticoids, 13 progestins, and 2 sterols, were validated and its quantitative data were visualized using hierarchically clustered heat maps to allow "steroid signatures". The devised method provided a good linearity (r 2 Ͼ 0.994) with the exception of cholesterol (r 2 ϭ 0.983). Precisions (% CV) and accuracies (% bias) ranged from 0.9% to 11.2% and from 92% to 119%, respectively, for most steroids tested. To evaluate metabolic changes, this method was applied to urine samples obtained from 59 patients with benign prostatic hyperplasia (BPH) versus 41 healthy male subjects. Altered concentrations of urinary steroids found and heat maps produced during this 70-compound study showed also differences between the ratios of steroid precursors and their metabolites (representing enzyme activity). Heat maps showed that oxidoreductases clustered (5␣-reductase, 3␣-HSD, 3␤-HSD, and 17␤-HSD, except for 20␣-HSD). These results support that data transformation is valid, since 5␣-reductase is a marker of BPH and 17␤-HSD is positively expressed in prostate cells. Multitargeted profiling analysis of steroids generated quantitative results that help to explain correlations between enzyme activities. The data transformation and visualization described may to be found in the integration with the mining biomarkers of hormone-dependent diseases. Many naturally occurring steroids with similar chemical structures could yield biological information [7]. Endogenous steroids are divided into five groups, namely, androgens, estrogens, corticoids, progestins, and sterols, which are generally synthesized from cholesterol in the adrenal cortex, ovaries, and testes (Scheme 1). In biosynthetic pathways of steroid hormone, two major types of enzymes are involved, cytochrome P450 and steroid oxidoreductase. Abnormalities of these enzymes often lead to hormonal imbalances that have serious consequences, and which are responsible for the development of hormone-dependent diseases (see Supplementary Table 1, which can be found in the electronic version of this article). For example, concentrations of corticoids and their metabolic ratios provide diagnostic evidence of apparent mineralocorticoid excesses caused by 11␤-HSD deficiency [8] and congenital adrenal hyperplasia, which are caused by deficiencies of enzymes like hydroxylase (at C-11, 17, and 21) or 3␤-HSD [9]. In addition, enhanced androgen activity generated by the conversion of testosterone to dihydrotestosterone (DHT) by 5␣-reductase was utilized to allow early therapeutic intervention in young men [10].Enzyme activity profiles can be used to describe the functional diversities of biological systems, which are d...

Bone morphogenetic protein (BMP) 7 counteracts physiological epithelial-to-mesenchymal transition, a process that is indicative of epithelial plasticity in developmental stages. Because epithelial-to-mesenchymal transition and its reversed process mesenchymal-to-epithelial transition (MET) are also involved in cancer progression, we investigated whether BMP7 plays a role in WM-266-4 melanoma cell growth and metastasis. An MTT assay was conducted in WM-266-4 and HEK293T cell lines to show the cell growth inhibition ability of BMP7 and cisplatin. Semiquantitative RT-PCR was used to determine MET in morphologically changed BMP7-treated melanoma cells. MET-induced cells expressed less a basic helix-loop-helix transcription factor (TWIST) in western blot analysis, and we confirm that BMP receptor (Alk2) siRNA transduction could restore TWIST protein expression via blocking of Smad 1, 5 and 8 signaling. Matrigel invasion and cell migration assays were done to investigate the BMP7-induced metastasis inhibition ability. BMP7 treatment only slightly reduced cell growth rate, but induced apparent MET. BMP7 also reduced the invasion and migration ability. Furthermore, BMP7 reduced the resistance of WM-266-4 cells to cisplatin. Collectively, our findings indicate that the metastatis inhibition ability of BMP7 is involved in MET, and that BMP7 could be used as a potential metastasis inhibitor in human melanoma cells. (Cancer Sci 2009; 100: 2218-2225 A lthough 90% of cancer deaths are caused by metastasis, (1) most chemotherapeutic agents cannot prevent tumor metastasis. The pathogenesis and mechanisms underlying this event are still poorly understood.(2-4) Metastasis is a 'hidden' event, which happens inside the body and is difficult to examine. It is believed to consist of four distinct steps: invasion, intravasation, extravasation, and metastatic colonization. Most carcinoma cells lose their cell-cell contact during the initial step of metastasis and move into the systemic circulation. For this, cells must acquire abilities of migration and invasion, together with cytoskeletal reorganization and active response to their microenvironment. In vivo video microscopy and quantitative approaches show that the first step, the acquisition of invasive ability and motility, is the rate-limiting step in the metastatic cascade (2,5) and clearly indicate that controlling this initial step of metastasis is critical for the development of novel strategies to prevent cancer metastasis. Epithelial-to-mesenchymal transition (EMT), a process vital for morphogenesis during embryonic development, is attracting attention as an important mechanism involved in the initial step of metastasis. During EMT, epithelial cells acquire fibroblast-like properties and show reduced intercellular adhesion and increased motility.(6,7) Conversely, mesenchymal cells possess remarkable plasticity and can eventually regain a fully differentiated epithelial phenotype via a mesenchymalto-epithelial transition (MET). (8,9) Because EMT involves the conversion of a sh...

PurposeDiagnosis and proper treatment of renal abscesses remains a challenge for physicians. We investigated the characteristics and comorbidity factors of renal abscesses measuring 5 cm or less and critically examined the effectiveness of conservative treatment.Materials and MethodsBetween February 2001 and March 2009 the records of 63 patients initially diagnosed at our hospital with renal or perirenal abscesses were retrospectively reviewed. In 63 patients with renal and perirenal abscesses, 51 abscesses measured 5 cm or less, and 49 abscesses were treated with intravenous antibiotics alone.ResultsMost patients were women (91.8%), and their mean age was 42.3 years. The mean size of renal abscesses was 3.6 cm. The most common predisposing condition was diabetes mellitus (DM) (46.9%). Common clinical features were fever (83.7%) and flank pain (53.1%). On urinalysis, 31 (64.6%) cases had positive bacterial cultures with Escherichia coli (50.0%) being the most common pathogen. All 49 patients were treated with broad-spectrum intravenous antibiotics alone. All patients showed complete clinical regression and resolution of the renal lesions shown by CT between 3 and 14 weeks. The average hospital stay was 15.3 days (range, 5-31 days). Significant predictors of a long hospital stay were age, abscess size, and DM.ConclusionMedium-sized as well as small-sized renal abscesses were treated successfully with intravenous antibiotics alone. DM was a significant predictor of prolonged hospital stay. If therapeutic drainage is believed to involve considerable risk, then intravenous antimicrobial therapy may be a good alternative treatment.