Dual blockade of BRD4 and ATR/WEE1 pathways exploits ARID1A loss in clear cell ovarian cancer.

ARID1A, an epigenetic tumor suppressor, is the most common gene mutation in clear-cell ovarian cancers (CCOCs). CCOCs are often resistant to standard chemotherapy and lack effective therapies. We hypothesized that ARID1A loss would increase CCOC cell dependency on chromatin remodeling and DNA repair pathways for survival. We demonstrate that combining BRD4 inhibitor (BRD4i) with DNA damage response inhibitors (ATR or WEE1 inhibitors; e.g. BRD4i-ATRi) was synergistic at low doses leading to decreased survival, and colony formation in CCOC in an ARID1A dependent manner. BRD4i-ATRi caused significant tumor regression and increased overall survival in ARID1AMUT but not ARID1AWT patient-derived xenografts. Combination BRD4i-ATRi significantly increased γH2AX, and decreased RAD51 foci and BRCA1 expression, suggesting decreased ability to repair DNA double-strand-breaks (DSBs) by homologous-recombination in ARID1AMUT cells, and these effects were greater than monotherapies. These studies demonstrate BRD4i-ATRi is an effective treatment strategy that capitalizes on synthetic lethality with ARID1A loss in CCOC.

High anti-Müllerian hormone levels might not reflect the likelihood of clinical pregnancy rate in IVF/ICSI treatment.

OBJECTIVE: To investigate if high anti-Müllerian hormone (AMH) concentration is a useful tool to predict the outcome of assisted reproductive treatment. METHODS: Retrospective cohort study involving 520 patients who underwent IVF/ICSI procedures in a university hospital. We measured the serum AMH level on day 3 of the menstrual cycle. Based on AMH levels, we divided the patients into three groups as follows: low (<25th percentile) AMH group, average (25th to 75th percentile) AMH group and high (>75th percentile) AMH group. We recorded the fertilization rate (FR), the number of oocytes retrieved, the number of good quality embryos (GQEs) and the clinical pregnancy rate (CPR). RESULTS: There was no difference between the three AMH groups in terms of maternal age, body mass index (BMI), follicle-stimulating hormone (FSH), estradiol (E2), luteinizing hormone (LH) and testosterone (T) in the IVF/ICSI cycles. The women in the high serum AMH group had a higher number of retrieved oocytes than those in the low or average AMH groups (p < 0.01) in the IVF/ICSI cycles. Compared with the low or average AMH groups, the women with high AMH levels had a higher number of good quality embryos (GQEs) in the IVF/ICSI cycles (p < 0.01). However, high AMH women had no significantly higher clinical pregnancy rate (CPR) compared to the women in the low or average AMH groups. In addition, for the prediction of CPR, the AMH levels alone were not an independent predictor of CPR for IVF and ICSI cycles in the ROC curve analysis. CONCLUSIONS: High anti-Müllerian hormone levels are an independent predictor of the number of retrieved oocytes and good quality embryos (GQEs), but might not reflect the likelihood of higher clinical pregnancy rates (CPR) in IVF/ICSI treatment.

Histological comparison of two special methods of staining melanin in human skin / Comparación histológica de dos métodos especiales de tinción de melanina en piel humana

SUMMARY: Human skin melanin was stained using the Fontana's silver nitrate method and Schmorl method. The results showed that, in the Fontana's silver nitrate method, melanin and silver-bound cells were black and other tissues were red. When stained using the Schmorl method, effects on melanin differed based on whether the nuclei were stained. When the nucleus was stained, melanin appeared blue-black or blue-green, and other tissue structures were purple. When the nucleus was not stained, melanin was orange and other structures were pink. Comparing the two staining methods, we concluded that Fontana's silver nitrate method takes a long time; in contrast, the Schmorl method showed two different types of results depending on whether the nucleus was stained, and it takes less time than Fontana staining, so we here consider the Schmorl method more suitable for special staining of melanin than Fontana's silver nitrate method.

RESUMEN: La melanina de la piel humana se tiñó utilizando el método del nitrato de plata de Fontana y el método Schmorl. Los resultados mostraron que, en el método del nitrato de plata de Fontana, la melanina y las células unidas a plata eran negras y otros tejidos eran rojos. Cuando se tiñó con el método de Schmorl, los efectos sobre la melanina difirieron en función de si se tiñeron los núcleos. Cuando se tiñó el núcleo, la melanina apareció de color azul-negro o azul-verde, y otras estructuras de tejido fueron de color púrpura. Cuando el núcleo no estaba teñido, la melanina era naranja y otras estructuras eran rosadas. Al comparar los dos métodos de tinción, llegamos a la conclusión de que el método del nitrato de plata de Fontana lleva mucho tiempo; por el contrario, el método Schmorl mostró dos tipos diferentes de resultados dependiendo de si el núcleo estaba teñido, y lleva menos tiempo que la tinción de Fontana, por lo que aquí consideramos que el método Schmorl es más adecuado para la tinción especial de melanina que el método del nitrato de plata de Fontana.

The effect of heavy metal contamination on the bacterial community structure at Jiaozhou Bay, China

Abstract In this study, determination of heavy metal parameters and microbiological characterization of marine sediments obtained from two heavily polluted sites and one low-grade contaminated reference station at Jiaozhou Bay in China were carried out. The microbial communities found in the sampled marine sediments were studied using PCR-DGGE (denaturing gradient gel electrophoresis) fingerprinting profiles in combination with multivariate analysis. Clustering analysis of DGGE and matrix of heavy metals displayed similar occurrence patterns. On this basis, 17 samples were classified into two clusters depending on the presence or absence of the high level contamination. Moreover, the cluster of highly contaminated samples was further classified into two sub-groups based on the stations of their origin. These results showed that the composition of the bacterial community is strongly influenced by heavy metal variables present in the sediments found in the Jiaozhou Bay. This study also suggested that metagenomic techniques such as PCR-DGGE fingerprinting in combination with multivariate analysis is an efficient method to examine the effect of metal contamination on the bacterial community structure.

The effect of heavy metal contamination on the bacterial community structure at Jiaozhou Bay, China

In this study, determination of heavy metal parameters and microbiological characterization of marine sediments obtained from two heavily polluted sites and one low-grade contaminated reference station at Jiaozhou Bay in China were carried out. The microbial communities found in the sampled marine sediments were studied using PCR-DGGE (denaturing gradient gel electrophoresis) fingerprinting profiles in combination with multivariate analysis. Clustering analysis of DGGE and matrix of heavy metals displayed similar occurrence patterns. On this basis, 17 samples were classified into two clusters depending on the presence or absence of the high level contamination. Moreover, the cluster of highly contaminated samples was further classified into two sub-groups based on the stations of their origin. These results showed that the composition of the bacterial community is strongly influenced by heavy metal variables present in the sediments found in the Jiaozhou Bay. This study also suggested that metagenomic techniques such as PCR-DGGE fingerprinting in combination with multivariate analysis is an efficient method to examine the effect of metal contamination on the bacterial community structure.(AU)

The effect of heavy metal contamination on the bacterial community structure at Jiaozhou Bay, China.

In this study, determination of heavy metal parameters and microbiological characterization of marine sediments obtained from two heavily polluted sites and one low-grade contaminated reference station at Jiaozhou Bay in China were carried out. The microbial communities found in the sampled marine sediments were studied using PCR-DGGE (denaturing gradient gel electrophoresis) fingerprinting profiles in combination with multivariate analysis. Clustering analysis of DGGE and matrix of heavy metals displayed similar occurrence patterns. On this basis, 17 samples were classified into two clusters depending on the presence or absence of the high level contamination. Moreover, the cluster of highly contaminated samples was further classified into two sub-groups based on the stations of their origin. These results showed that the composition of the bacterial community is strongly influenced by heavy metal variables present in the sediments found in the Jiaozhou Bay. This study also suggested that metagenomic techniques such as PCR-DGGE fingerprinting in combination with multivariate analysis is an efficient method to examine the effect of metal contamination on the bacterial community structure.

Biotransformation of isoimperatorin by rat liver microsomes and its quantification by LC-MS/MS method.

The aim of the present research was to establish a comprehensive strategy to identify the metabolites of isoimperatorin after biotransformation with rat liver microsomes in vitro, and further describe metabolic kinetic characteristics of isoimperatorin and its main metabolites. Utilizing liquid chromatography with time of flight mass spectrometry (LC-TOF-MS), 18 metabolites (M 1-18) were characterized according to the typical fragment ions and literature data. Among them, M-2, 3, 5, 9, 10, and 15 were new compounds. To further verify structures of the metabolites, five main metabolites were obtained from the magnifying biotransformation incubation system, and their chemical structures were elucidated as 8-hydroxyoxypeucedanin (M-3), hydroxypeucedanin hydrate (M-4), E-5-(4-hydroxy-3-methyl-2-alkenyloxy)-psoralen (M-11), Z-5-(4-hydroxy-3-methyl-2-alkenyloxy)-psoralen (M-12), and oxypeucedanin (M-16) by various spectroscopy methods including IR, MS and NMR. A simple new liquid chromatography with triple quadrupole tandem mass spectrometry (LC-QqQ-MS) method was developed for the simultaneous determination of isoimperatorin and its main metabolites. The analysis was performed on a Diamonsil™ ODS C18 column with acetonitrile-water containing 0.1% formic acid as mobile phase. Total run time was 20.0 min. The results suggested that the method we exhibited was successfully applied for analysis of isoimperatorin and its metabolites. The study provides essential data for proposing metabolite pathway and further pharmacological study of isoimperatorin.

Venezuelines A-G, new phenoxazine-based alkaloids and aminophenols from Streptomyces venezuelae and the regulation of gene target Nur77.

Five new phenoxazine-based alkaloids venezuelines A-E (1-5) and two new aminophenols venezuelines F-G (6-7), as well as three known analogues exfoliazone, chandrananimycin D and carboxyexfoliazone were isolated from the fermentation broth of the marine-derived bacterium Streptomyces venezuelae. The structures of new compounds were determined on the basis of extensive spectroscopic analysis. The cytotoxic activity of these compounds against a panel of tumor cell lines were tested, while the regulation of gene target Nur77 of 2 and exfoliazone (8) were evaluated.