An alternative miRISC targets a cancer-associated coding sequence mutation in FOXL2.

Recent evidence suggests that animal microRNAs (miRNAs) can target coding sequences (CDSs); however, the pathophysiological importance of such targeting remains unknown. Here, we show that a somatic heterozygous missense mutation (c.402C>G; p.C134W) in FOXL2, a feature shared by virtually all adult-type granulosa cell tumors (AGCTs), introduces a target site for miR-1236, which causes haploinsufficiency of the tumor-suppressor FOXL2. This miR-1236-mediated selective degradation of the variant FOXL2 mRNA is preferentially conducted by a distinct miRNA-loaded RNA-induced silencing complex (miRISC) directed by the Argonaute3 (AGO3) and DHX9 proteins. In both patients and a mouse model of AGCT, abundance of the inversely regulated variant FOXL2 with miR-1236 levels is highly correlated with malignant features of AGCT. Our study provides a molecular basis for understanding the conserved FOXL2 CDS mutation-mediated etiology of AGCT, revealing the existence of a previously unidentified mechanism of miRNA-targeting disease-associated mutations in the CDS by forming a non-canonical miRISC.

miR-22-3p and miR-30e-5p Are Associated with Prognosis in Cervical Squamous Cell Carcinoma.

Alteration in expression of miRNAs can cause various malignant changes and the metastatic process. Our aim was to identify the miRNAs involved in cervical squamous cell carcinoma (SqCC) and metastasis, and to test their utility as indicators of metastasis and survival. Using microarray technology, we performed miRNA expression profiling on primary cervical SqCC tissue (n = 6) compared with normal control (NC) tissue and compared SqCC that had (SqC-M; n = 3) and had not (SqC-NM; n = 3) metastasized. Four miRNAs were selected for validation by qRT-PCR on 29 SqC-NM and 27 SqC-M samples, and nine metastatic lesions (ML-SqC), from a total of 56 patients. Correlation of miRNA expression and clinicopathological parameters was analyzed to evaluate the clinical impact of candidate miRNAs. We found 40 miRNAs differentially altered in cervical SqCC tissue: 21 miRNAs were upregulated and 19 were downregulated (≥2-fold, p < 0.05). Eight were differentially altered in SqC-M compared with SqC-NM samples: four were upregulated (miR-494, miR-92a-3p, miR-205-5p, and miR-221-3p), and four were downregulated (miR-574-3p, miR-4769-3p, miR-1281, and miR-1825) (≥1.5-fold, p < 0.05). MiR-22-3p might be a metastamiR, which was gradually further downregulated in SqC-NM > SqC-M > ML-SqC. Downregulation of miR-30e-5p significantly correlated with high stage, lymph node metastasis, and low survival rate, suggesting an independent poor prognostic factor.

Concurrent activation of OsAMT1;2 and OsGOGAT1 in rice leads to enhanced nitrogen use efficiency under nitrogen limitation.

Nitrogen (N) is a major factor for plant development and productivity. However, the application of nitrogenous fertilizers generates environmental and economic problems. To cope with the increasing global food demand, the development of rice varieties with high nitrogen use efficiency (NUE) is indispensable for reducing environmental issues and achieving sustainable agriculture. Here, we report that the concomitant activation of the rice (Oryza sativa) Ammonium transporter 1;2 (OsAMT1;2) and Glutamate synthetase 1 (OsGOGAT1) genes leads to increased tolerance to nitrogen limitation and to better ammonium uptake and N remobilization at the whole plant level. We show that the double activation of OsAMT1;2 and OsGOGAT1 increases plant performance in agriculture, providing better N grain filling without yield penalty under paddy field conditions, as well as better grain yield and N content when plants are grown under N llimitations in field conditions. Combining OsAMT1;2 and OsGOGAT1 activation provides a good breeding strategy for improving plant growth, nitrogen use efficiency and grain productivity, especially under nitrogen limitation, through the enhancement of both nitrogen uptake and assimilation.

Synergism of AZD6738, an ATR Inhibitor, in Combination with Belotecan, a Camptothecin Analogue, in Chemotherapy-Resistant Ovarian Cancer.

Epithelial ovarian cancer remains the leading cause of mortality among all gynecologic malignancies owing to recurrence and ultimate development of chemotherapy resistance in the majority of patients. In the chemotherapy-resistant ovarian cancer preclinical model, we investigated whether AZD6738 (an ataxia telangiectasia and Rad3-related (ATR) inhibitor) could synergize with belotecan (a camptothecin analog and topoisomerase I inhibitor). In vitro, both chemotherapy-resistant and chemotherapy-sensitive ovarian cancer cell lines showed synergistic anti-proliferative activity with a combination treatment of belotecan and AZD6738. The combination also demonstrated synergistic tumor inhibition in mice with a chemotherapy-resistant cell line xenograft. Mechanistically, belotecan, a DNA-damaging agent, increased phospho-ATR (pATR) and phospho-Chk1 (pChk1) in consecutive order, indicating the activation of the DNA repair system. This consequently induced G2/M arrest in the cell cycle analysis. However, when AZD6738 was added to belotecan, pATR and pChk1 induced by belotecan alone were suppressed again. A cell cycle analysis in betotecan showed a sub-G1 increase as well as a G2/M decrease, representing the release of G2/M arrest and the induction of apoptosis. In ascites-derived primary cancer cells from both chemotherapy-sensitive and -resistant ovarian cancer patients, this combination was also synergistic, providing further support for our hypothesis. The combined administration of ATR inhibitor and belotecan proved to be synergistic in our preclinical model. This combination warrants further investigation in a clinical trial, with a particular aim of overcoming chemotherapy resistance in ovarian cancer.

Development and Validation of a Method for Determining the Quercetin-3-O-glucuronide and Ellagic Acid Content of Common Evening Primrose (Oenothera biennis) by HPLC-UVD.

Toward the standardization of common evening primrose (Oenothera biennis) sprout extract (OBS-E), we aimed to obtain indicator compounds and use a validated method. HPLC-UVD allowed simultaneous quantification of the indicator compounds quercetin-3-O-glucuronide and ellagic acid. The method was validated in terms of specificity, linearity, precision, accuracy, and limit of detection/limit of quantification (LOD/LOQ). High specificity and linearity was demonstrated, with correlation coefficients of 1.0000 for quercetin-3-O-glucuronide and 0.9998 for ellagic acid. The LOD/LOQ values were 0.486/1.472 µg/mL for quercetin-3-O-glucuronide and 1.003/3.039 µg/mL for ellagic acid. Intra-day and inter-day variability tests produced relative standard deviation for each compound of <2%, a generally accepted precision criterion. High recovery rate were also obtained, indicating accuracy validation. The OBS-E prepared using various concentrations of ethanol were then analyzed. The 50% ethanol extract had highest content of quercetin-3-O-glucuronide, whereas the 70% ethanol extract possessed the lowest. However, the ellagic acid content was highest in the 70% ethanol extract and lowest in the 90% ethanol extract. Thus, quercetin-3-O-glucuronide and ellagic acid can be used industrially as indicator compounds for O. biennis sprout products, and our validated method can be used to establish indicator compounds for other natural products.

Large-Scale Assembly of Peptide-Based Hierarchical Nanostructures and Their Antiferroelectric Properties.

An effective strategy is developed to create peptide-based hierarchical nanostructures through the meniscus-driven self-assembly in a large area and fabricate antiferroelectric devices based on these nanostructures for the first time. The diphenylalanine hierarchical nanostructures (FF-HNs) are self-assembled by vertically pulling a substrate from a diphenylalanine (FF) solution dissolved in a miscible solvent under precisely controlled conditions. Owing to the unique structural properties of FF nanostructures, including high crystallinity and α-helix structures, FF-HNs possess a net electrical dipole moment, which can be switched in an external electric field. The mass production of antiferroelectric devices based on FF-HNs can be successfully achieved by means of this biomimetic assembly technique. The devices show an evident antiferroelectric to ferroelectric transition under dark conditions, while the ferroelectricity is found to be tunable by light. Notably, it is discovered that the modulation of antiferroelectric behaviors of FF-HNs under glutaraldehyde exposure is due to the FF molecules that are transformed into cyclophenylalanine by glutaraldehyde. This work provides a stepping stone toward the mass production of self-assembled hierarchical nanostructures based on biomolecules as well as the mass fabrication of electronic devices based on biomolecular nanostructures for practical applications.

Control of Epitaxial BaFe2As2 Atomic Configurations with Substrate Surface Terminations.

Atomic layer controlled growth of epitaxial thin films of unconventional superconductors opens the opportunity to discover novel high temperature superconductors. For instance, the interfacial atomic configurations may play an important role in superconducting behavior of monolayer FeSe on SrTiO3 and other Fe-based superconducting thin films. Here, we demonstrate a selective control of the atomic configurations in Co-doped BaFe2As2 epitaxial thin films and its strong influence on superconducting transition temperatures by manipulating surface termination of (001) SrTiO3 substrates. In a combination of first-principles calculations and high-resolution scanning transmission electron microscopy imaging, we show that Co-doped BaFe2As2 on TiO2-terminated SrTiO3 is a tetragonal structure with an atomically sharp interface and with an initial Ba layer. In contrast, Co-doped BaFe2As2 on SrO-terminated SrTiO3 has a monoclinic distortion and a BaFeO3- x initial layer. Furthermore, the superconducting transition temperature of Co-doped BaFe2As2 ultrathin films on TiO2-terminated SrTiO3 is significantly higher than that on SrO-terminated SrTiO3, which we attribute to shaper interfaces with no lattice distortions. This study allows the design of the interfacial atomic configurations and the effects of the interface on superconductivity in Fe-based superconductors.

Therapeutic effects of endoscopic ablation in patients with Hunner type interstitial cystitis.

OBJECTIVE: To investigate the efficacy of endoscopic ablation of Hunner lesions (HLs) in patients with interstitial cystitis (IC) and to find predictors of early recurrence of HLs. PATIENTS AND METHODS: A prospective study was performed of patients with Hunner type IC who underwent transurethral ablation. We repeated endoscopic ablation when symptoms and HLs recurred during the follow-up period. The primary endpoint was recurrence-free time. Secondary endpoints were a change in frequency, nocturia, and urgency episodes, and changes in visual analogue scale (VAS) pain scores and other symptom indices at follow-up visits. RESULTS: A total of 72 patients were analysed. The median (range) follow-up period was 29.5 (12.0-50.0) months. After primary ablation treatment, HLs recurred in 75.0% (54/72) of patients, and the median (sd) recurrence-free time was 12.0 (1.6) months. Amongst the 54 patients with recurrence, 50 underwent a second ablation treatment. HLs occurred in 44.0% (22/50) of individuals after the second operation, and the median (sd) recurrence-free time was 18.0 (5.1) months. Lower maximal cystometric capacity (odds ratio 1.01, 95% confidence interval 1.001-1.013) was the predictive factor for early recurrence. There were significant improvements in the VAS pain scores, O'Leary-Sant Interstitial Cystitis Symptom Index and Problem Index, Pelvic Pain and Urgency/Frequency Patient Symptom Scale after treatment (all, P < 0.05). CONCLUSIONS: Endoscopic ablation is an effective treatment option for HLs and significantly reduces pain and improves voiding symptoms. Repeat ablation upon recurrence could help symptom control and bladder preservation only if the bladder capacity is maintained.

Diagnosing bladder outlet obstruction using the penile cuff test in men with lower urinary tract symptoms.

AIM: The current standard diagnostic test to evaluate bladder outlet obstruction (BOO) is pressure-flow study (PFS). The penile cuff test (PCT) was introduced as a non-invasive alternative to PFS to determine the isovolumetric bladder pressure and also flow rate. The aim of the study was to evaluate the diagnostic accuracy and acceptability of the PCT, compared to those of PFS, in the assessment of BOO in men. METHODS: A total of 146 consecutive men with an International Prostate Symptom Score (IPSS) >12 and lower urinary tract symptoms (LUTS) for >6 months were included in this single-institution, prospective, diagnostic study. The primary outcome was to evaluate the diagnostic accuracy of PCT. Secondary outcomes included pain severity measured on a visual analog scale (VAS), procedure time, and adverse events. In addition, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were estimated. RESULTS: Based on the PFS results, the sensitivity of PCT was 89.7%, the PPV 54.2%, the specificity 71.8%, and the NPV 94.9%. The mean pain VAS for the PFS and PCT were 5.04 ± 2.17 and 1.83 ± 1.98, respectively (P < 0.0001). The mean procedure time for PCT (6.3 ± 0.6 min) was shorter than that of PFS (23.3 ± 2.2 min) (P < 0.0001). There were no adverse events reported during PCT. CONCLUSIONS: Given its high NPV, PCT may be an efficient screening test for BOO in men. In addition, PCT is advantageous over PFS with regard to its short procedure time and acceptable tolerability.

Long-term outcomes of primary implantation and revisions of artificial urinary sphincter in men with stress urinary incontinence.

AIMS: To evaluate long-term outcomes of primary implantation and revisions of artificial urinary sphincter (AUS) in men with stress urinary incontinence (SUI). METHODS: The medical records of 155 consecutive patients who underwent AMS 800™ implantation from 2003 to 2015 were reviewed. Treatment success (TS) was defined as no need for pads, and social continence (SC) was defined as a need for pad ≤1/day at last follow-up. The efficacy, complication rates, and associated risk factors with reoperation and durability of primary AUS implantation, as well as treatment outcomes following AUS revision were evaluated. RESULTS: The median age was 69.0 years (range 27-85), and median follow-up was 45.1 months (range 9-126). The rates of TS and SC were 63.2% and 84.5%, respectively. The reoperation rate of AUS was 26.4%. Non-mechanical failure (70.7%) was a dominant etiology of reoperation. The 5-year device survival rate without reoperation was 67.0%. The immediate TS rates of primary AUS without reoperation (pAUS) and AUS revision without secondary reoperation (rAUS) groups were 88.6% and 79.2% (P = 0.352), respectively. At last follow-up, the SC rate was higher in patients with pAUS (92.1%) than with rAUS (62.5%) (P = 0.001). CONCLUSIONS: Although there are appreciable rates of reoperation, the AMS 800™ offers high rates of continence in men with SUI. The immediate TS rate was comparable in patients with pAUS and rAUS. At the final follow-up, however, the SC rate was significantly higher in patients with pAUS than with rAUS.

Imprint Control of BaTiO3 Thin Films via Chemically Induced Surface Polarization Pinning.

Surface-adsorbed polar molecules can significantly alter the ferroelectric properties of oxide thin films. Thus, fundamental understanding and controlling the effect of surface adsorbates are crucial for the implementation of ferroelectric thin film devices, such as ferroelectric tunnel junctions. Herein, we report an imprint control of BaTiO3 (BTO) thin films by chemically induced surface polarization pinning in the top few atomic layers of the water-exposed BTO films. Our studies based on synchrotron X-ray scattering and coherent Bragg rod analysis demonstrate that the chemically induced surface polarization is not switchable but reduces the polarization imprint and improves the bistability of ferroelectric phase in BTO tunnel junctions. We conclude that the chemical treatment of ferroelectric thin films with polar molecules may serve as a simple yet powerful strategy to enhance functional properties of ferroelectric tunnel junctions for their practical applications.

Notch3-specific inhibition using siRNA knockdown or GSI sensitizes paclitaxel-resistant ovarian cancer cells.

Notch signaling plays an important role in ovarian cancer chemoresistance, which is responsible for recurrence. Gamma-secretase inhibitor (GSI) is a broad-spectrum Notch inhibitor, but it has serious side effects. The efficacy of Notch3-specific inhibition in paclitaxel-resistant ovarian cancers was assessed in this study, which has not yet been evaluated relative to GSI. To analyze the effect of Notch3-specific inhibition on paclitaxel-resistant ovarian cancers, we compared cell viability, apoptosis, cell migration, angiogenesis, cell cycle, and spheroid formation after treatment with either Notch3 siRNA or GSI in paclitaxel-resistant SKpac cells and parental SKOV3 cells. Expression levels of survival, cell cycle, and apoptosis-related proteins were measured and compared between groups. Notch3 was significantly overexpressed in chemoresistant cancer tissues and cell lines relative to chemosensitive group. In paclitaxel-resistant cancer cells, Notch inhibition significantly reduced viability, migration, and angiogenesis and increased apoptosis, thereby boosting sensitivity to paclitaxel. Spheroid formation was also significantly reduced. Both Notch3 siRNA-treated cells and GSI-treated cells arrested in the G2/M phase of the cell cycle. Proteins of cell survival, cyclin D1 and cyclin D3 were reduced, whereas p21 and p27 were elevated. Both GSI and Notch3 siRNA treatment reduced expression of anti-apoptotic proteins (BCL-W, BCL2, and BCL-XL) and increased expression of pro-apoptotic proteins (Bad, Bak, Bim, Bid, and Bax). These results indicate that Notch3-specific inhibition sensitizes paclitaxel-resistant cancer cells to paclitaxel treatment, with an efficacy comparable to that of GSI. This approach would be likely to avoid the side effects of broad-spectrum GSI treatment. © 2015 Wiley Periodicals, Inc.

Comparison of the efficacy and safety of tolterodine 2 mg and 4 mg combined with an α-blocker in men with lower urinary tract symptoms (LUTS) and overactive bladder: a randomized controlled trial.

OBJECTIVE: To evaluate the efficacy and safety of low-dose (2 mg) tolterodine extended release (ER) with an α-blocker compared with standard-dose (4 mg) tolterodine ER with an α-blocker for the treatment of men with residual storage symptoms after α-blocker monotherapy. PATIENTS AND METHODS: The study was a 12-week, single-blind, randomized, parallel-group, non-inferiority trial that included men with residual storage symptoms despite receiving at least 4 weeks of α-blocker treatment. Inclusion criteria were total International Prostate Symptom Score (IPSS) ≥12, IPSS quality-of-life item score ≥3, and ≥8 micturitions and ≥2 urgency episodes per 24 h. The primary outcome was change in the total IPSS score from baseline. Bladder diary variables, patient-reported outcomes and safety were also assessed. RESULTS: Patients were randomly assigned to addition of either 2 mg tolterodine ER (n = 47) or 4 mg tolterodine ER (n = 48) to α-blocker therapy for 12 weeks. Patients in both treatment groups had a significant improvement in total IPSS score (-5.5 and -6.3, respectively), micturition per 24 h (-1.3 and -1.7, respectively) and nocturia per night (-0.4 and -0.4, respectively). Changes in IPSS, bladder diary variables, and patient-reported outcomes were not significantly different between the treatment groups. All interventions were well tolerated by patients. CONCLUSIONS: These results suggest that 12 weeks of low-dose tolterodine ER add-on therapy is similar to standard-dose tolterodine ER add-on therapy in terms of efficacy and safety for patients experiencing residual storage symptoms after receiving α-blocker monotherapy.

Drug persistence and compliance affect patient-reported outcomes in overactive bladder syndrome.

PURPOSE: To evaluate persistence and compliance for patients receiving antimuscarinics for overactive bladder (OAB), and to assess patient-reported outcomes (PROs) according to persistence and compliance. METHODS: This was a 24-week, multicenter, prospective, observational study that included 952 OAB patients who had newly started antimuscarinics. Patients aged ≥18 years with a total OAB Symptom Score (OABSS) ≥3 and an urgency score of OABSS ≥2 were eligible for the study. Drug persistence and compliance were evaluated at 4, 12, and 24 weeks. Changes in scores on PROs were compared between groups (persistence vs. non-persistence and compliance vs. non-compliance) after 24 weeks. Factors contributing to persistence were examined using multivariate logistic regression. RESULTS: After 24 weeks, 56.8 % of patients remained on treatment. The persistence rates were 85.6 and 71.4 % after 4 and 12 weeks, respectively. The compliance rates were 75.6, 53.8, and 34.3 % after 4, 12, and 24 weeks, respectively. Patients who were persistent in taking antimuscarinics resulted in significant improvements in OABSS and OAB questionnaire short form score compared with those who were non-persistent (all p < 0.05). Changes from baseline in OABSS (p = 0.735) and the EuroQoL five-dimensions score (p = 0.384) were not significantly different between compliant and non-compliant groups. Predictors of high persistence included older age (OR 1.017, p = 0.007) and dry OAB (OR 1.422, p = 0.013). CONCLUSIONS: Patients who were persistent with antimuscarinics showed significant improvements in PROs compared to those who were non-persistent.

Diffusion-weighted magnetic resonance imaging for prediction of insignificant prostate cancer in potential candidates for active surveillance.

OBJECTIVES: To investigate whether the apparent diffusion coefficient (ADC) from diffusion-weighted magnetic resonance imaging (DW-MRI) could help improve the prediction of insignificant prostate cancer in candidates for active surveillance (AS). METHODS: Enrolled in this retrospective study were 287 AS candidates who underwent DW-MRI before radical prostatectomy. Patients were stratified into two groups; Group A consisted of patients with no visible tumour or a suspected tumour ADC value > 0.830 × 10(-3) mm(2)/sec and Group B consisted of patients with a suspected tumour ADC value < 0.830 × 10(-3) mm(2)/sec. We compared pathological outcomes in each group. RESULTS: Group A had 243 (84.7 %) patients and Group B had 44 (15.3 %) patients. The proportion of organ-confined Gleason ≤ 6 disease and insignificant prostate cancer was significantly higher in Group A than Group B (61.3 % vs. 38.6 %, p = 0.005 and 47.7 % vs. 25.0 %, p = 0.005, respectively). On multivariate analysis, a high ADC value was the independent predictor of organ-confined Gleason ≤ 6 disease and insignificant prostate cancer (odds ratio = 2.43, p = 0.011 and odds ratio = 2.74, p = 0.009, respectively). CONCLUSION: Tumour ADC values may be a useful marker for predicting insignificant prostate cancer in candidates for AS. KEY POINTS: • ADC from DW-MRI can help assess prostate cancer aggressiveness in potential AS candidates. • There was a closed correlation between higher ADC values and insignificant prostate cancer. • The absence of lesions on DWI/DWI can help select potential AS candidates.

Surgical outcome of a repeat midurethral sling procedure after failure of a first procedure.

INTRODUCTION AND HYPOTHESIS: Although there is no consensus on the management of persistent or recurrent stress urinary incontinence (SUI) after placement of a midurethral synthetic sling (MUS), a repeat MUS procedure is commonly performed with favorable results. The aim of this study was to evaluate the efficacy of a repeat MUS procedure compared to the primary procedure in women with SUI, and to investigate factors associated with the failure of the repeat procedure. METHODS: We retrospectively analyzed data from 53 women who underwent a repeat MUS procedure and 102 women who underwent a primary MUS procedure at a single center. Success was defined as no urine leakage during physical activity based on the Sandvik questionnaire. Outcomes were assessed using the Sandvik Severity Index and Incontinence-Quality of Life (I-QOL) questionnaire. Multivariate logistic regression analysis was used to determine the factors predicting failure of the repeat procedure. RESULTS: The success rate was 76.5 % for the primary MUS procedure (78/102 patients) and 69.8 % for the repeat MUS procedure (37/53 patients; p = 0.369). The mean follow-up duration was significantly longer for the primary procedure (83.8 months vs. 54.6 months, p < 0.001). SUI and all domain scores of the I-QOL were significantly better following the repeat MUS procedure than following the primary procedure. In the multivariate analysis, SUI grade 3 was the only independent factor predicting failure of the repeat qq (odds ratio 7.610, p = 0.023). CONCLUSIONS: A repeat MUS procedure after a failed primary MUS procedure was shown to be effective. However, a repeat procedure may be unsuccessful in patients with grade 3 SUI.

SUVmax of 18F-FDG PET/CT in the differential diagnosis of benign and malignant thyroid nodules according to tumor volume.

BACKGROUND: The aim of this study was to investigate whether the maximum standardized uptake value (SUVmax) measured on fluorine-18 fluorodeoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) could be used as the primary means of differential diagnosis of thyroid nodules when tumor volume is assessed. METHODS: We studied 192 patients who underwent preoperative 18F-FDG PET/CT followed by thyroidectomy. We evaluated the correlation between the volume of thyroid nodules, 18F-FDG uptake on visual analysis, and the mean SUVmax measured on 18F-FDG PET/CT. RESULTS: When stratified by tumor volume, the mean SUVmax was higher in malignant than in benign nodules in nodules≥1 cm3 (p<0.001). However, it did not differ between benign and malignant nodules smaller than 1 cm3. At a cut-off value of SUVmax of 6, the respective sensitivities of 18F-FDG PET/CT, ultrasonography, and fine needle aspiration cytology were 60.8, 96.4, and 99.1%, and the respective specificities were 95.9, 98.2, and 96.8%. CONCLUSIONS: 18F-FDG PET/CT is limited as a primary modality in the differential diagnosis of benign and malignant thyroid nodules because of its low sensitivity.

MicroRNA expression profiling and Notch1 and Notch2 expression in minimal deviation adenocarcinoma of uterine cervix.

BACKGROUND: MicroRNA (miRNA) expression is known to be deregulated in cervical carcinomas. However, no data is available about the miRNA expression pattern for the minimal deviation adenocarcinoma (MDA) of uterine cervix. We sought to detect deregulated miRNAs in MDA in an attempt to find the most dependable miRNA or their combinations to understand their tumorigenesis pathway and to identify diagnostic or prognostic biomarkers. We also investigated the association between those miRNAs and their target genes, especially Notch1 and Notch2. METHODS: We evaluated miRNA expression profiles via miRNA microarray and validated them using.real-time PCR assays with 24 formalin-fixed, paraffin-embedded tissue blocks of MDA and 11 normal proliferative endocervical tissues as control. Expression for Notch1 and 2 was assessed by immunohistochemistry. RESULTS: MiRNA-135a-3p, 192-5p, 194-5p, and 494 were up-regulated, whereas miR-34b-5p, 204-5p, 299-5p, 424-5p, and 136-3p were down-regulated in MDA compared with normal proliferative endocervical tissues (all P<0.05). Considering the second-order Akaike Information Criterion consisting of likelihood ratio and number of parameters, miR-34b-5p showed the best discrimination power among the nine candidate miRNAs. A combined panel of miR-34b-5p and 194-5p was the best fit model to discriminate between MDA and control, revealing 100% sensitivity and specificity. Notch1 and Notch2, respective target genes of miR-34b-5p and miR-204-5p, were more frequently expressed in MDA than in control (63% vs. 18%; 52% vs. 18%, respectively, P<0.05). MiR-34b-5p expression level was higher in Notch1-negative samples compared with Notch1-positive ones (P<0.05). Down-regulated miR-494 was associated with poor patient survival (P=0.036). CONCLUSIONS: MDA showed distinctive expression profiles of miRNAs, Notch1, and Notch2 from normal proliferative endocervical tissues. In particular, miR-34b-5p and 194-5p might be used as diagnostic biomarkers and miR-494 as a prognostic predictor for MDA. The miR-34b-5p/Notch1 pathway as well as Notch2 might be important oncogenic contributors to MDA.

Quantitative determination of enzalutamide, an anti-prostate cancer drug, in rat plasma using liquid chromatography-tandem mass spectrometry, and its application to a pharmacokinetic study.

This report details a method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) that allows one to determine the concentration of an atypical anticancer drug, enzalutamide, in rat plasma. Specifically, this method involves the addition of an acetonitrile and bicalutamide (internal standard) solution to plasma samples. Following centrifugation of this mixture, an aliquot of the supernatant was directly injected into the LC-MS/MS system. Separation was achieved using a column packed with octadecylsilica (5 µm, 2.1 × 50 mm) with 10 mM ammonium acetate in acetonitrile as the mobile phase; detection was accomplished using MS/MS by multiple-reaction monitoring via an electrospray ionization source. This method demonstrated a linear standard curve (r = 0.997) over a concentration range of 0.001-1 µg/mL, as well as an intra- and inter-assay precision of 2.7 and 5.1%, respectively, and an accuracy range from 100.8 to 105.6%. The lower limit of quantification was 1.0 ng/mL in 50 µL of rat plasma sample. We also demonstrated that this analytical method could be successfully applied to the pharmacokinetic study of enzalutamide in rats.