Designing a novel photoinduced electron transfer-based small-molecule fluorescent probe specific for CYP3A isozymes.

Cytochrome P450 (CYPs) are oxidoreductases distributed in various tissues in plants and animals. Among the CYP families, CYP3A is the most abundant in vivo, particularly in humans, and it is involved in the metabolism of many drugs. It is crucial to measure CYP3A activity for both pharmaceuticals and agrochemicals because inhibition or induction of this enzyme can seriously affect the occurrence of toxicity or efficacy. In the present study, a novel fluorescent probe, 6-(2,5-bis(trifluoromethyl)benzyloxy)-9-(4-methoxy-2-methylphenyl)-3H-xanthen-3-one (BMX, quantum efficiency: 21%), was designed and synthesized. The design was done by photoinduced electron transfer strategy. BMX was specifically metabolized only using CYP3A to generate 2-Me-4-MeO TokyoGreen (quantum efficiency: 85%), resulting in strong fluorescence in the presence of CYP3A isozymes. Protein assays using recombinant human, rat, and mouse CYP isozymes demonstrated the selective metabolism of BMX and production of fluorescence only by CYP3A in all species.

Below-the-elbow intervention for Buerger's disease.

An increasing attention has been paid to endovascular therapy for lower limb ischemia in patients with Buerger's disease. However, critical hand ischemia in Buerger's disease patients has been underappreciated despite a tremendous advancement of endovascular therapy for peripheral arterial disease. Herein, we describe endovascular "hand" salvage with a below-the-elbow intervention.

Effect of single tibial artery revascularization on microcirculation in the setting of critical limb ischemia.

BACKGROUND: Benefits of 2-dimensional (2D) angiosome-oriented infrapopliteal revascularization remain controversial. The aim of this retrospective study was to clarify the effect of single tibial artery revascularization on the dorsal and plantar microcirculation of critically ischemic limbs based on skin perfusion pressure (SPP). METHODS AND RESULTS: Fifty-seven interventions that only involved either anterior tibial artery (ATA) or posterior tibial artery (PTA) revascularization were included in this study. SPP was measured on the dorsal side (theoretically ATA perfusion area) and the plantar side (theoretically PTA perfusion area) before and after the procedure. Dorsal and plantar SPP increased significantly, from 33 (IQR 23-40.5) to 52 (IQR 32.5-65) mm Hg (P<0.0001) and 31.6±16.1 to 44.8±19.2 mm Hg (P=0.001) after ATA revascularization, respectively, and from 29.3±14.0 to 42.4±19.7 mm Hg (P=0.003) and 29.3±9.8 to 43.5±15.9 mm Hg (P<0.001) after PTA revascularization, respectively. Both ATA and PTA revascularization were not associated with any significant differences in ΔSPP between the dorsal and the plantar regions of the foot. Only 64% and 58% of ATA revascularization cases showed higher post-SPP and ΔSPP on the dorsal side than on the plantar side, respectively. Also, only 47% and 40% of PTA revascularization cases showed higher post-SPP and ΔSPP on the plantar side than on the dorsal side, respectively. CONCLUSIONS: Single tibial artery revascularization, whether of the ATA or PTA, yielded comparable improvements in microcirculation of the dorsal and plantar foot. Approximately half of the feet revascularized had a change in microcirculation that was not consistent with the 2D angiosome theory.

Vasospastic limb ischemia presenting acute and chronic limb ischemia.

Vasospastic limb ischemia might have been underappreciated compared to vasospasm in other territories such as heart and brain. However, an increasing awareness of this vascular disorder can be translated to an improved patients' care. Herein, we report a case of vasospasm presenting acute and chronic limb ischemia in four extremities.

Contemporary crossing techniques for infrapopliteal chronic total occlusions.

The synergism of technical refinement and advanced technology has significantly increased the popularity of infrapopliteal intervention. Since chronic total occlusion (CTO) is a common disorder among patients with symptomatic infrapopliteal artery disease, infrapopliteal CTO intervention is now evolving rapidly in the field of endovascular intervention. Guidewire crossing through the CTO is essential for a successful procedure. We review up-to-date infrapopliteal CTO crossing techniques based on the current literature.

Impact of aortorenal morphology on renal artery stent procedures: significance of aortic tortuosity and renal artery derivation.

PURPOSE: To clarify the impact of aortorenal morphology on renal artery stenting procedures. METHODS: A retrospective study evaluated 142 consecutive renal artery stenting procedures performed for de novo atherosclerotic renal artery stenosis in 119 patients (62 men; mean age 72±9 years, range 41-93). All procedures were done via a transfemoral approach without distal protection. Aortorenal morphology was classified into 3 types based on the relationship between abdominal aortic tortuosity and renal artery derivation. Using a straight reference line centered on the most angulated point of the inner curve of the infrarenal abdominal aorta, type 1 referred to a renal artery ostium that was more than half of the aortic diameter distance from the reference line in the greater curvature and less than half in the lesser curvature. Type 2 referred to a renal artery ostium that was less than half of the aortic diameter distant from the reference line in the greater curvature and more than half in the lesser curvature. Type 3 referred to a renal artery ostium that was beyond the reference line in the greater curvature or more than one aortic diameter from the reference line in the lesser curvature. The technical success rate, procedure time, final engagement technique, shape of the guide catheter used, and any adverse events were analyzed. RESULTS: Type 1 aortorenal morphology was observed in 91 cases, type 2 in 30, and type 3 in 21. All cases were successfully treated; there were no technical complications, in-hospital cardiovascular events, or deaths. Procedure time differed significantly (p<0.001) among the 3 types (type 1: 19.6±5.6 minutes, type 2: 23.3±6.8 minutes, and type 3: 32.3±9.6 minutes; p<0.05 for type 1 vs. 2, p<0.001 for type 2 vs. 3, and p<0.001 for type 1 vs. 3). There were also significant differences among types in terms of engagement technique and guide catheter shape. CONCLUSION: Aortorenal morphology was significantly associated with procedure time and the selection of engagement technique and guide catheter shape.

Endovascular recanalization of failed distal bypass in Buerger's disease.

In the treatment of Buerger's disease, bypass surgery with the use of an autologous vein has served as a treatment option in cases in which distal target vessel has been available. However, failed bypass occlusion can result in a devastating clinical scenario. Herein, we report a successful endovascular revascularization of failed distal bypass graft as a last resort for a patient with Burger's disease with ischemic rest pain and extensive tissue loss.

A novel selective androgen receptor modulator, NEP28, is efficacious in muscle and brain without serious side effects on prostate.

Age-related androgen depletion is known to be a risk factor for various diseases, such as osteoporosis and sarcopenia. Furthermore, recent studies have demonstrated that age-related androgen depletion results in accumulation of ß-amyloid protein and thereby acts as a risk factor for the development of Alzheimer's disease. Supplemental androgen therapy has been shown to be efficacious in treating osteoporosis and sarcopenia. In addition, studies in animals have demonstrated that androgens can play a protective role against Alzheimer's disease. However, androgen therapy is not used routinely for these indications, because of side effects. Selective androgen receptor modulators (SARMs) are a new class of compounds. SARMs maintain the beneficial effects of androgens on bone and muscle while reducing unwanted side effects. NEP28 is a new SARM exhibiting high selectivity for androgen receptor. To investigate the pharmacological effects of NEP28, we compared the effects on muscle, prostate, and brain with mice that were androgen depleted by orchidectomy and then treated with either placebo, NEP28, dihydrotestosterone, or methyltestosterone. We demonstrated that NEP28 showed tissue-selective effect equivalent to or higher than existing SARMs. In addition, the administration of NEP28 increased the activity of neprilysin, a known Aß-degrading enzyme. These results indicate that SARM is efficacious for the treatment of not only osteoporosis and sarcopenia, but also Alzheimer's disease.

Subintimal angioplasty of lengthy femorotibial total occlusion in Buerger's disease.

PURPOSE: To report successful subintimal angioplasty of a lengthy femorotibial occlusion in a patient with Buerger's disease, with wound healing and limb salvage. CASE REPORT: A 38-year-old female heavy smoker was referred to our hospital for treatment of extensive infectious tissue loss, with severe foot pain 1 month after early failure of a distal bypass graft. Angiography revealed total occlusion in the femoropopliteal and infrapopliteal arteries. Endovascular recanalization was attempted in order to establish "straight-line flow" to the foot on the verge of limb loss. The subintimal angioplasty technique with a 0.014-inch hydrophilic guidewire facilitated successful crossing of the occlusive femoropopliteal and posterior tibial arteries. The lesions were serially dilated (standard and cutting balloons). Angiography demonstrated antegrade flow to the foot without flow-limiting dissection, and the serious pain dramatically disappeared. Complete wound healing was observed 5 months after initial revascularization with the assistance of repeat angioplasty for restenosis. CONCLUSION: Contemporary endovascular therapy using the subintimal angioplasty technique could represent a viable option for Buerger's disease.

Discovery of potent and orally bioavailable 17ß-hydroxysteroid dehydrogenase type 3 inhibitors.

We have previously reported the discovery of a new class of potent inhibitors of 17ß-hydroxysteroid dehydrogenase type 3 (17ß-HSD3) derived from benzylidene oxazolidinedione and thiazolidinedione scaffolds. In this study, these analogs were designed, synthesized, and evaluated in a human cell-based assay. The detailed structure-activity relationship (SAR) surrounding this pharmacophore were developed, and consequently a number of compounds from this series demonstrated single-digit nanomolar 17ß-HDS3 inhibitory activity in vitro. Subsequent optimization work in pursuit of the improvement of oral bioavailability demonstrated in vivo proof-of-concept by prodrug strategy based on phosphate esters for these 17ß-HSD3 inhibitors. When a phosphate ester 16 was administered orally at a high dose of 100mg/kg, 16 showed approximately two times more potent testosterone (T)-lowering effect against a positive control in the luteinizing hormone-releasing hormone (LH-RH)-induced T production assay. The T-lowering effect continued at ca 10% level of control over 4h after administration. The nonsteroidal molecules based on this series have the potential to provide unique and effective clinical opportunities for treatment of prostate cancer.

Identification of oxazolidinediones and thiazolidinediones as potent 17ß-hydroxysteroid dehydrogenase type 3 inhibitors.

Novel and potent inhibitors of 17ß-hydroxysteroid dehydrogenase type 3 (17ß-HSD3) were identified based on oxazolidinedione and thiazolidinedione derivatives, starting from a high-throughput screening hit, 5-(3-bromo-4-hydroxybenzyl)-3-(4-methoxyphenyl)-1,3-thiazol-2-one. 5-(3-Bromo-4-hydroxybenzylidene)-3-(4-methoxyphenyl)-2-thioxo-1,3-thiazolidin-4-one exhibited a promising activity profile and demonstrated significant selectivity over the related 17ß-HSD isoenzymes and nuclear receptors.

Coumarins as novel 17beta-hydroxysteroid dehydrogenase type 3 inhibitors for potential treatment of prostate cancer.

The synthesis and SAR studies of 3- and 4-substituted 7-hydroxycoumarins as novel 17beta-HSD3 inhibitors are discussed. The most potent compounds from this series exhibited low nanomolar inhibitory activity with acceptable selectivity versus other 17beta-HSD isoenzymes and nuclear receptors.

Crossveinless-2 controls bone morphogenetic protein signaling during early cardiomyocyte differentiation in P19 cells.

Increasing evidence indicates that bone morphogenetic proteins (BMPs) are crucial for cardiac induction, specification, and development. Although signaling of BMPs is tightly regulated through soluble BMP-binding proteins, how they regulate BMP signaling during cardiac differentiation remains unknown. To identify molecules responsible for BMP signaling during early cardiomyocyte differentiation of P19 cells, cDNA subtraction was performed. We found a bimodal expression of the BMP-binding protein Crossveinless-2 (Cv2) during cardiomyocyte differentiation; Cv2 is temporally expressed earlier than cardiac transcription factors such as Nkx2.5 and Tbx5 and acts as a suppressor for BMP signaling in P19 cells. We established a P19 clonal cell line harboring a cardiac alpha-myosin heavy chain promoter-driven enhanced green fluorescent protein gene to monitor cardiac differentiation by flow cytometry. Treatment with BMP2 during the first 2 days of differentiation suppressed cardiomyocyte differentiation through activation of down-stream targets Smad1/5/8 protein and Id1 gene, whereas treatment with Cv2 conversely inhibited Smad1/5/8 activation and Id1 expression, leading to increased generation of cardiac cells. RNA interference-mediated knockdown (KD) of endogenous Cv2 showed increased Smad1/5/8 activation and impaired cardiomyocyte differentiation. Expression of cardiac mesoderm markers was reduced, whereas expression of Id1 and endoderm markers such as Sox7, Hnf4, and E-cadherin was induced in Cv2-kinase dead cells. These phenotypes were rescued by the addition of Cv2 protein to the culture media during the first 2 days of differentiation or co-culture with parental cells. These data suggest that Cv2 may specify cardiac mesodermal lineage through inhibition of BMP signaling at early stage of cardiogenesis.