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Hyperactivation of ribosome biogenesis leads to hepatocyte transformation and plays pivotal roles in hepatocellular carcinoma (HCC) development. We aimed to identify critical ribosome biogenesis proteins that are overexpressed and crucial in HCC progression. HEAT repeat containing 1 (HEATR1) expression and clinical correlations were analyzed using The Cancer Genome Atlas and Gene Expression Omnibus databases and further evaluated by immunohistochemical analysis of an HCC tissue microarray. Gene expression was knocked down by small interfering RNA. HEATR1 -knockdown cells were subjected to viability, cell cycle, and apoptosis assays and used to establish subcutaneous and orthotopic tumor models. Chromatin immunoprecipitation and quantitative polymerase chain reaction were performed to detect the association of candidate proteins with specific DNA sequences. Endogenous coimmunoprecipitation combined with mass spectrometry was used to identify protein interactions. We performed immunoblot and immunofluorescence assays to detect and localize proteins in cells. The nucleolus ultrastructure was detected by transmission electron microscopy. Click-iT (Thermo Fisher Scientific) RNA imaging and puromycin incorporation assays were used to measure nascent ribosomal RNA and protein synthesis, respectively. Proteasome activity, 20S proteasome foci formation, and protein stability were evaluated in HEATR1 -knockdown HCC cells. HEATR1 was the most up-regulated gene in a set of ribosome biogenesis mediators in HCC samples. High expression of HEATR1 was associated with poor survival and malignant clinicopathologic features in patients with HCC and contributed to HCC growth in vitro and in vivo. HEATR1 expression was regulated by the transcription factor specificity protein 1, which can be activated by insulin-like growth factor 1–mammalian target of rapamycin complex 1 signaling in HCC cells. HEATR1 localized predominantly in the nucleolus, bound to ribosomal DNA, and was associated with RNA polymerase I transcription/processing factors. Knockdown of HEATR1 disrupted ribosomal RNA biogenesis and impaired nascent protein synthesis, leading to reduced cytoplasmic proteasome activity and inhibitory-κB/nuclear factor-κB signaling. Moreover, HEATR1 knockdown induced nucleolar stress with increased nuclear proteasome activity and inactivation of the nucleophosmin 1-MYC axis. Our study revealed that HEATR1 is up-regulated by insulin-like growth factor 1–mammalian target of rapamycin complex 1–specificity protein 1 signaling in HCC and functions as a crucial regulator of ribosome biogenesis and proteome homeostasis to promote HCC development. [Display omitted] Nucleolar protein HEAT repeat containing 1 is up-regulated by mechanistic target of rapamycin complex 1 signaling and contributes to hepatocellular carcinoma progression through dominating ribosome biogenesis and proteome homeostasis. [ABSTRACT FROM AUTHOR]

• A new diphenylethane (1) and a new dibenz[ b , f ]oxepin (2) were isolated from the stems and leaves of Dioscorea oppositifolia L. • Compounds 1 , 3 , 7 , 9 , and 10 exhibted estrogenic activity at doses of 10 μ M were mediated by ER β. • The diphenylethanes might belong to undescribed phytoestrogens. A new diphenylethane, 2′,4-dihydroxy-2,5-dimethoxybibenzyl (1), and a new dibenz[ b,f ]oxepin, 10,11-dihydro-2-methoxydibenz[ b,f ]oxepin-4-ol (2), as well as eight known compounds (3 – 10) were isolated from the stems and leaves of Dioscorea oppositifolia L. Their structures were elucidated by detailed analysis of comprehensive spectroscopic data. Estrogenic activity of all isolated compounds were evaluated using a proliferation assay in MCF-7 cells. Compounds 1 , 3 , 7 , 9 , and 10 increased the proliferation of MCF-7 cell and the expression of ER β in the MCF-7 cell line. In addition, blockage of ER β completely abolished cell proliferation and expression of ER β in MCF-7cells, suggesting that the proliferation effects of Compounds 1 , 3 , 7 , 9 , and 10 were ER β -mediated. The diphenylethanes might belong to undescribed phytoestrogens. [ABSTRACT FROM AUTHOR]

• CREB1 was expressed in granulosa cells and theca cells of mature follicles. • Methylation of DMR of promoter were increased in HP group than that in LP group. • The non-CGI methylation of CREB1 promoter may regulate gene expression on sheep ovaries. This study aimed to investigate the effect of different methylated regions of cyclic-AMP response element binding protein 1 (CREB1) by comparing the high prolificacy (HP) group and low prolificacy (LP) group, which was detected in our previous study. The expression level of CREB1 mRNA in the ovaries of the HP group was higher than in the LP group (P < 0.05). The differential methylated region (DMR) had 4 methylated CG dinucleotides(CGs): −1546, −1544, −1494 and −1464. The DNA methylation levels of -1546 CGs and −1464 CGs were significantly higher in the HP group than in the LP group (P < 0.05). The activity from −1296 to +26 (without DMR) was significantly higher than the activity from −1598 to +26 (with DMR) (P < 0.05). The result of 5-aza-2′-deoxycytidine treatment indicated that the inhibition DNA methylation of DMR reduced the transcription of CREB1. The bioinformatics predictive analysis were found that the −1546 CG site was located in the CCAAT/enhancer-binding protein alpha (CEBPA) binding site and the -1464 CG site was located in the Sp1 binding site. Finally, this study revealed the relationship between the methylation of non-CpG sites of the promoter and transcription of CREB1. This study will provide a theoretical basis of the Hu sheep ovaries associated with DNA methylation. [ABSTRACT FROM AUTHOR]

Abstract: As-prepared single-wall carbon nanotubes (SWCNTs) usually aggregate into bundles that have diameters of tens of nanometers, and these bundles are very difficult to separate by post-synthesis treatment for characterization and applications. In this study, we applied a constant electric field during the synthesis of SWCNTs by arc discharge method. As a result, ∼98% of the SWCNT bundles obtained have diameters less than 10nm, and more than 50% of the bundles are less than 3nm in diameter, indicating a large population of isolated single or pairs of SWCNTs. [Copyright &y& Elsevier]

Abstract: Three new ursane-type triterpenes, glutinosalactone A–C (1–3), were isolated from the 50% aqueous acetone extract of the leaves of Rehmannia glutinosa. Their structures were elucidated on the basis of spectral analysis (IR, NMR and MS spectroscopy). The cytotoxic effects of compounds 1–3 against three human cancer cell lines (MCF-7, MG63 and HepG2) were also evaluated. Compound 3 showed cytotoxic activities with IC50 values of 8.35–39.25μM. [Copyright &y& Elsevier]

Abstract: The lung-stage schistosomulum has been regarded as the main target of protective immunity induced by radiation-attenuated vaccines (RAV) in the mouse model of schistosomiasis, and immune mechanisms mediated by the CD4+ Th1 response play a major role in the RAV model. To identify Th1 epitopes rapidly within molecules from the lung schistosomulum of Schistosoma japonicum, in the present study we analyzed transcriptome data from normal and radiation-attenuated lung schistosomula of S. japonicum and Schistosoma mansoni. We selected six genes with high levels of expression of their transcripts as sample sequences from the lung schistosomula. From these six sequences, by using different algorithms, we predicted six promiscuous Th cell epitopes that are capable of binding to both murine and human MHC class II molecules. To validate our in silico prediction experimentally, first, the gene expressions of the six sequences in day 3 lung-stage schistosomula were assessed using reverse-transcription PCR (polymerase chain reaction) analysis. The result showed that all six sequences predicted can be expressed in normal day 3 schistosomula. Second, we measured the direct binding of the four peptides predicted above to APCs (Antigen Presenting Cells) from the BALB/c mouse strain using a fluorometric method, and found that the four peptides could bind to both I-Ad and I-Ed molecules of the mice. Finally, the proliferation and profiles of cytokine production by spleen lymphocytes from the BALB/c mice immunized with the six predicted peptides were detected in vitro using modified MTT (Methyl Thiazolyl Tetrazolium), and flow cytometry methods, respectively. The results showed that three of the six predicted peptides could induce a recall CD4+ Th1 response in vitro. These results demonstrate that potential Th1-type epitopes can be identified rapidly by a combination of in silico analysis of transcriptomes of lung-stage schistosomula with experimental validation. [Copyright &y& Elsevier]

Abstract: The objective was to investigate the effects of the transgenic donor cell synchronization method, oocyte sources, and other factors, on production of hLF-gene nucleus transfer dairy goats. Three transfected cell lines from ear biopsies from three 3-mo-old Saanen dairy goats (designated Number 1, Number 2, and Number 3, respectively) were selected as karyoplast donors for somatic cell nuclear transfer (SCNT) after detailed identification (including PCR and sequencing of PCR products). In donor cell cycle synchronization studies, the apoptosis rate of hLF transgenic fibroblasts was not different (P > 0.05) after 3 days of serum starvation or 2 days of contact inhibition. Additionally, there was no effect (P > 0.05) on developmental capacity of reconstructed embryos; however, the kidding rate of recipients in the serum starvation group was higher than that in the contact inhibition group (18 vs. 0%, respectively). The production efficiency of the transgenic cloned goats using donor cells from the Number 1 dairy goat cell line was higher than those using the Number 2 and the Number 3 cell lines (kidding rates were 18, 2, and 0%, respectively, P < 0.05). The oocyte source did not significantly affect the pregnancy rate of hLF-transgenic cloned dairy goats, but more fetuses were aborted when using in vitro matured oocytes compared to in vivo matured oocytes. In summary, utilizing transfected 3-mo-old dairy goat fibroblasts as donor cells, seven live offspring were produced, and the hLF gene was successfully integrated. This study provided additional insights into preparation of donor cells and recipient oocytes for producing transgenic cloned goats through SCNT. [Copyright &y& Elsevier]

In this letter, we investigate the general rational solutions and breather solutions of a higher-order integrable nonlinear Schrödinger (NLS) equation based on Darboux transformation (DT). The rational solutions including W-shape traveling wave solution that is not reported and rogue wave solution are constructed. Time-periodic Kuznetsov–Ma breather, space-periodic Akhmediev breather and time–space periodic breather solutions are obtained. Besides, the interaction properties of two breather solutions are also displayed through numerical simulation. The results exhibit the new dynamical properties in extended nonlinear integrable physical models. [ABSTRACT FROM AUTHOR]

Background & Aims Agents designed to block or alter cytokinesis can kill or stop proliferation of cancer cells. We aimed to identify cytokinesis-related proteins that are overexpressed in hepatocellular carcinoma (HCC) cells and might be targeted to slow liver tumor growth. Methods Using the Oncomine database, we compared the gene expression patterns in 16 cancer microarray datasets and assessed gene enrichment sets using gene ontology. We performed immunohistochemical analysis of an HCC tissue microarray and identified changes in protein levels that are associated with patient survival times. Candidate genes were overexpressed or knocked down with small hairpin RNAs in SMMC7721, MHCC97H, or HCCLM3 cell lines; we analyzed their proliferation, viability, and clone-formation ability and their growth as subcutaneous or orthotopic xenograft tumors in mice. We performed microarray analyses to identify alterations in signaling pathways and immunoblot and immunofluorescence assays to detect and localize proteins in tissues. Yeast 2-hybrid screens and mass spectrometry combined with co-immunoprecipitation experiments were used to identify binding proteins. Protein interactions were validated with co-immunoprecipitation and proximity ligation assays. Chromatin immunoprecipitation, promoter luciferase activity, and quantitative real-time polymerase chain reaction analyses were used to identify factors that regulate transcription of specific genes. Results The genes that were most frequently overexpressed in different types of cancer cells were involved in cell division processes. We identified 3 cytokinesis-regulatory proteins among the 10 genes most frequently overexpressed by all cancer cell types. Rac GTPase activating protein 1 (RACGAP1) was the cytokinesis-regulatory protein that was most highly overexpressed in multiple cancers. Increased expression of RACGAP1 in tumor tissues was associated with shorter survival times of patients with cancer. Knockdown of RACGAP1 in HCC cells induced cytokinesis failure and cell apoptosis. In microarray analyses, we found knockdown of RACGAP1 in SMMC7721 cells to reduce expression of genes regulated by yes-associated protein (YAP) and WW domain containing transcription regulator 1 (WWTR1 or TAZ). RACGAP1 reduced activation of the Hippo pathway in HCC cells by increasing activity of RhoA and polymerization of filamentous actin. Knockdown of YAP reduced phosphorylation of RACGAP1 and redistribution at the anaphase central spindle. We found transcription of the translocated promoter region, nuclear basket protein (TPR) to be regulated by YAP and coordinately expressed with RACGAP1 to promote proliferation of HCC cells. TPR redistributed upon nuclear envelope breakdown and formed complexes with RACGAP1 during mitosis. Knockdown of TPR in HCC cells reduced phosphorylation of RACGAP1 by aurora kinase B and impaired their redistribution at the central spindle during cytokinesis. STAT3 activated transcription of RACGAP in HCC cells. Conclusions In an analysis of gene expression patterns of multiple tumor types, we found RACGAP1 to be frequently overexpressed, which is associated with shorter survival times of patients. RACGAP1 promotes proliferation of HCC cells by reducing activation of the Hippo and YAP pathways and promoting cytokinesis in coordination with TPR. Graphical abstract [ABSTRACT FROM AUTHOR]

Electronic structure and magnetoelectric coupling effect for M/BiFeO 3 (M = Co, Fe) multiferroic superlattices are predicted by first-principles within the GGA + U. Hybridizations of O-2p and M-3d orbital are found on the interface of superlattices. The charge redistribution and ferroelectric polarization make magnetic moment different between the bottom and top interface, which results in the ME coupling effect. The magnetoelectric coupling coefficient of Co/BiFeO 3 is bigger than that of Fe/BiFeO 3 by comparing with their ferroelectric and magnetic properties. Electronic structure and magnetoelectric coupling effect for M/BiFeO 3 (M = Co, Fe) multiferroic superlattices are predicted by first-principles within the GGA + U. Image 1 • Hybridization and charge transfer between interfaces of M/BiFeO 3 superlattices are found. • Magnetoelectric coupling coefficient of Co/BiFeO 3 is bigger than that of Fe/BiFeO 3 multiferroic superlattices. • The electronic structure and magnetic properties of system have been calculated by the first-principles. [ABSTRACT FROM AUTHOR]

Maternal prenatal hypoxia is an important contributor to intrauterine growth restriction (IUGR), which impedes fetal lung maturation and leads to the development of chronic lung diseases. Although evidence suggests the involvement of pyroptosis in IUGR, the molecular mechanism of pyroptosis is still unclear. Nuclear factor erythroid 2-related factor 2 (Nrf2) has been found to potentially interact with gasdermin D (GSDMD), the key protein responsible for pyroptosis, indicating its crucial role in inhibiting pyroptosis. Therefore, we hypothesized that Nrf2 deficiency is a key molecular responsible for lung pyroptosis in maternal hypoxia-induced IUGR offspring mice. Pregnant WT and Nrf2-/- mice were exposed to hypoxia (10.5 % O 2) to mimic IUGR model. We assessed body weight, lung histopathology, pulmonary angiogenesis, oxidative stress levels, as well as mRNA and protein expressions related to inflammation in the 2-week-old offspring. Additionally, we conducted a dual-luciferase reporter assay to confirm the targeting relationship between Nrf2 and GSDMD. Our findings revealed that offspring with maternal hypoxia-induced IUGR exhibited reduced birth weight, catch-up growth delay, and pulmonary dysplasia. Furthermore, we observed impaired nuclear translocation of Nrf2 and increased GSDMD-mediated pyroptosis in these offspring with IUGR. Moreover, the dual-luciferase reporter assay demonstrated that Nrf2 could directly inhibit GSDMD transcription; deficiency of Nrf2 exacerbated pyroptosis and pulmonary dysplasia in offspring with maternal hypoxia-induced IUGR. Collectively, our findings suggest that Nrf2 deficiency induces GSDMD-mediated pyroptosis and pulmonary dysplasia in offspring with maternal hypoxia-induced IUGR; thus highlighting the potential therapeutic approach of targeting Nrf2 for treating prenatal hypoxia-induced pulmonary dysplasia in offspring. [Display omitted] • Antenatal maternal hypoxia triggers GSDMD-mediated pyroptosis and results in pulmonary dysplasia in IUGR offspring. • Nrf2 nuclear translocation is decreased in lung tissues of IUGR offspring. • Nrf2 inhibits GSDMD transcription and its deficiency aggravated pyroptosis and pulmonary dysplasia in IUGR offspring. [ABSTRACT FROM AUTHOR]

ATP is essential for mammalian sperm to maintain fertilizing capacity. Metformin (Met) can activate 5′-AMP-activated protein kinase (AMPK) to maintain energy homeostasis. Thus, the aim of the present study was to investigate whether Met can improve testis function, semen quality, antioxidant and autophagy capacity through AMPK mediation of energy metabolism in goats. Twelve adult goats were randomly divided into three dietary treatments. All goats were fed a basal diet for 3 weeks and then assigned to a Met supplementation diet containing 0, 150, or 300 mg/kg for 8 weeks. The results showed that sperm viability, sperm membranal functional integrity, and acrosome integrity increased (P < 0.05) relative to the other treatments in the 300 mg/kg Met group. Growth hormone (GH) and insulin-like growth factor (IGF-1) in the 300 mg/kg Met group significantly decreased (P < 0.05) relative to the control group. Estrogen levels (E 2) in the 300 mg/kg Met group remarkably improved (P < 0.05) compared with the control group. The activities of the antioxidant enzymes catalase (CAT), glutathione peroxidase (GSH-px), and superoxide dismutase (SOD) significantly increased (P < 0.05) in the 300 mg/kg Met group relative to the control group. A significant increase in AMPK and p-AMPK protein expression in the 300 mg/kg Met group was observed relative to the control group (P < 0.05). Belicin-1 and LC3II/I protein expression was significantly increased by adding Met to the diet (P < 0.05) and reached a maximum in the 300 mg/kg Met group. In addition, differentially expressed genes (DEGs) of goat testis were confirmed by RNA-seq. GO enrichment analysis revealed that DEGs were enriched in testicular metabolism and sperm development-related functional pathways. Overall, the results indicate that Met may play an important role in the regulation of testis function, semen quality, antioxidant, and autophagy capacity. These findings will help elucidate the role of Met in goat testis development. • The addition of Met improves reproductive performance of goats, significantly improved semen acrosome integrity and plasma membrane integrity. • The expression of AMPK and p-AMPK proteins were significantly increased. • The autophagy marker proteins and the activities of antioxidant enzymes were significantly increased. [ABSTRACT FROM AUTHOR]

Abstract: Oocyte aging severely decreases the quality of oocytes, which hampers fertilization and subsequent embryo development. In the present study, age-dependent molecular changes in goat oocytes were investigated. First, the quality of goat oocytes with various in vitro culture times (24, 30, 36, 48, and 60 hours) was evaluated on the basis of developmental rates of parthenogenetically activated embryos and apoptosis of cumulus cells (CCs). Second, relative gene expression of six genes (mitochondrial genes: PGC-1α and NRF-1; epigenetic modification genes: SNRPN and HAT1; mitotic spindle checkpoint protein: SMAD2; and hyaluronan synthase gene: HAS3) were analyzed during oocyte aging. Third, we further studied the changes of seven genes (PGC-1α and NRF-1; apoptotic-related genes: BAX and BCL2; hyaluronan synthase gene: HAS2; metabolism-related gene: STAR; and superoxide dismutase gene: SOD1) in CCs during oocyte aging. In these studies, the blastocyst rate gradually decreased and the number of apoptotic cells significantly increased as the culture time increased (P < 0.05). Moreover, relative gene expressions of PGC-1α, NRF-1 and SMAD2 significantly decreased from 24 to 36 hours (P < 0.05), whereas the levels of HAT1 and HAS3 slowly increased as culture was prolonged. Furthermore, the levels of PGC-1α, BCL2, HAS2 and SOD1 quickly reduced, and BAX significantly increased from 24 to 36 hours in aged CCs (P < 0.05). In conclusion, goat oocytes started to age at 30 hours in vitro culture, and gene expression patterns of oocytes and CCs significantly changed as the oocytes aged. Gene expression pattern changes in CCs may provide a convenient and effective way to detect oocyte aging without compromising oocyte integrity. [Copyright &y& Elsevier]

Abstract: It has not so far been possible to identify rapidly and effectively the anti-schistosomiasis Th cell epitopes that are capable of simulating IFN-γ (Interferon-gamma)-mediated Th1-type protective immunity in response to radiation-attenuated schistosome cercaria. With the advance of the omics studies of schistosomes, an approach that used reverse vaccinology probably resolved the above problems. In this “proof-of-principle” study, first, we selected 31 secreted or transmembrane protein sequences sampled from sequences of the transcriptome of Schistosoma japonicum, and analyzed characteristics of these proteins by using conventional bioinformatics tools. Second, putative promiscuous Th cell epitopes within these proteins were predicted using three to four different immuno-informatics algorithms for the prediction of MHC (Major Histocompatibility Complex) class-II binding peptides. We predicted using these in silico approaches promiscuous Th cell epitopes that are capable of binding to both murine and human MHC class-II molecules. To validate our in silico prediction experimentally, BALB/c mice were immunized with the five predicted peptides, and the proliferative responses and cytokine production of lymphocytes from the immunized BALB/c mice were assessed in vitro by modified MTT (Methyl Thiazolyl Tetrazolium), ELISA (Enzyme-linked Immunosorbent Assay) and flow cytometry methods. The results showed that two of the five predicted peptides could induce a Th1-type response in vitro. These results suggest that promiscuous Th1 cell epitopes from secreted or transmembrane proteins of S. japonicum can be identified using a strategy of reverse vaccinology. [Copyright &y& Elsevier]

Abstract: A series of novel phenacylimidazolium derivatives, bearing an aryl or alkyl substituent at position-1 and a phenacyl substituent at position-3 of the imidazole ring, has been prepared and evaluated in vitro against a panel of human tumor cell lines. Phenacylimidazolium bromides bearing a highly sterically hindered aryl group at position-1 and an electron-rich phenacyl or naphthylacyl substituent at position-3 of imidazole ring proved to be more active than imidazolium bromides with other substituted groups. In particular, compound 5j was found to be the most potent compounds with IC50 values lower than 5.0μM against 8 strains human tumor cell lines and more active than cisplatin (DDP). [Copyright &y& Elsevier]

Abstract: Parkinson’s disease is the second most common neurodegenerative disorders after Alzheimer’s disease in the elderly. Abundant evidence showed that proinflammatory factors were involved in the pathogenesis of sporadic Parkinson’s disease (SPD). Interleukin-1 (IL-1) is a cytokine that plays an important role in neurodegenerative disease. Previous association studies between genetic polymorphisms of IL-1 alpha and PD have showed conflicting results and no such study was done in Chinese. We recruited 533 SPD patients and 530 controls in Chinese Han population to investigate the association of IL-1α C-899T allele and risk for PD. Real-time PCR was used to detect the polymorphism, and multiple logistic regression, Chi square test and survival analysis were performed to explore the association. The distribution of IL-1α alleles was significantly different between the cases and controls, and the T allele was associated with a reduced risk of PD (OR: 0.72, 95%CI: 0.54–0.97, ρ =0.033). However, survival analysis showed that the T allele did not delay the onset age of PD (T allele vs. non-T allele log rank: χ 2 =0.14, p =0.70). Our data suggest that the T allele carriers have less inclination to have PD in Chinese Han population. [Copyright &y& Elsevier]

Nanos3 is from a highly conserved family of genes with putative RNA-binding activity and plays an important role in germ cell development. Yet, its function in Leydig cells (LCs) for supporting germ cell development, is still poorly understood. In this study, we investigated the expression pattern and function of Nanos3 in the sheep testis. The Nanos3 mRNA and protein were highly expressed in the ram testis. The sheep Nanos3 amino acid sequence was 93.6%–88.6% homologous with those of other mammalian species. The expression of Nanos3 in the ram testis was significantly lower in 9 and 24-month-old sheep than in at 3-month-old sheep (P < 0.05). Immunohistochemistry analysis revealed that sheep Nanos3 was not only localized at spermatogenesis cell but also at LCs. Thus the functions of Nanos3 in LCs were further investigated by using siRNA-Nanos3 in vitro. The secretion of T hormone and expression of testosterone synthesis-related enzymes in LCs were significantly inhibited (P < 0.05) by siRNA-Nanos3. Moreover, the BAX/Bcl-2 mRNA and protein levels were significantly increased (P < 0.05) by siRNA-Nanos3. In addition, we examined the expression levels of Nanos3 mRNA and protein in energy-restricted(ER) rams and in vitro ER LCs. Their levels were also significantly decreased (P < 0.05) and were accompanied by an increased BAX/Bcl-2 mRNA expression compared with the control group (P < 0.05). To the best of our knowledge, our study is the first to show that Nanos3 is an important regulator of testosterone biosynthesis in rams for supporting germ cell development. • The secretion of T hormone and testosterone synthesis-related enzymes in LCs were inhibited by siRNA-Nanos3. • The BAX/Bcl-2 mRNA and protein increased by inhibiting Nanos3. • Nanos3 regulates the apoptosis of the LCs. [ABSTRACT FROM AUTHOR]

Peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PPARGC1A) acts as a powerful coactivator of many transcriptional factors that relate to granulosa cell (GC) apoptosis. In this study, the miRNAs mediating goat follicular atresia and luteinized granulosa cell (LGC) apoptosis induced by hydrogen peroxide (H 2 O 2) via PPARGC1A were investigated. Our results showed that miR-1197–3p targeted PPARGC1A was predicted by bioinformatics algorithm and verified by luciferase reporter assay. In addition, miR-1197–3p promoted goat LGC apoptosis via PPARGC1A through mitochondrial-dependent apoptosis pathway, and these effects could be restored by PPARGC1A overexpression. Moreover, H 2 O 2 -induced LGC apoptosis significantly upregulated miR-1197–3p expression and downregulated PPARGC1A level. Pretreatment of miR-1197–3p inhibitor alleviated LGC apoptosis induced by 400 μM H 2 O 2 for 12 h, and preserved the mitochondrial membrane potential by increasing PPARGC1A expression. In conclusion, miR-1197–3p might act as an essential regulator of goat LGC apoptosis potentially via the mitochondrial-dependent apoptosis pathway by targeting PPARGC1A. • miR-1197–3p promotes goat LGC apoptosis via directly targeting PPARGC1A. • 400 μM H 2 O 2 for 12 h treatment was the optimal condition for H 2 O 2 induced goat LGC apoptosis model. • Suppression of miR-1197–3p attenuated H 2 O 2 -induced LGC apoptosis by targeting PPARGC1A. [ABSTRACT FROM AUTHOR]

Seven new lignanamides, lyciumamides D–J ( 1 – 4 and 9 – 11 ), together with nine known analogues ( 5 – 8 and 12 – 16 ), were isolated from the root bark of Lycium chinense . The structures of the isolated compounds were elucidated on the basis of NMR spectroscopic and HRESIMS data. All isolated compounds were evaluated for antihyperlipidemic activities in HepG2 cells. The primary structure-activity relationships were discussed. [ABSTRACT FROM AUTHOR]

A flavanone C-glycoside, steppogenin-5′-C- β -D-glucopyranoside, six prenylated 2-arylbenzofuran derivatives, moracin O-3″-O- β -D-glucopyranoside, moracin O-3′-O- β -D-xylopyranoside, moracin P-2″-O- β -D-glucopyranoside, moracin P-3′-O- β -D-glucopyranoside, moracin P-3′-O- α -L-arabinopyranoside and moracin P-3′-O-[ β -D-glucopyranosyl-(1 → 2)]- α -L-arabinopyranoside, two phenolic acids, 2,4-dihydroxy-5-(4-hydroxybenzyl) benzoic acid and 2,4-dihydroxy-5-(3,4-dihydroxybenzyl) benzoic acid, as well as three known compounds, moracinoside C, moracin O, and moracin P were isolated from the root bark of Morus alba L. Their structures were ascertained on the basis of spectroscopic evidence. The protective effects of the compounds against doxorubicin-induced cardiomyopathy in H9c2 cells was investigated in vitro . Of all of the isolated compounds, moracin P-3′-O- β -D-glucopyranoside, moracin O and moracin P had a strong protective influence against doxorubicin-induced cell death with EC 50 values of 9.5 ± 2.6, 4.5 ± 1.3, and 8.8 ± 2.4 μM, respectively. [ABSTRACT FROM AUTHOR]

Fiber tractography techniques in diffusion magnetic resonance imaging have become a primary tool for studying the fiber architecture of biological tissues both noninvasively and in vivo. Streamline tracking, as a simple and efficient tractography technique, is widely used to reconstruct fiber pathways. It is however very sensitive to noisy estimation of local fiber orientations. In this paper, we propose a bundle constrained streamline method to accurately reconstruct multifiber pathways. The method introduces neighboring fiber consistency constraint in the tracking process and reconstructs fiber pathways that have optimal tradeoff between consistency with local fiber orientation estimations and similarity with neighboring fiber segment orientations. Results on synthetic, physical phantom and real human brain DW images show that the proposed method allows regular fiber pathways to be reconstructed and outperforms existing techniques. [ABSTRACT FROM AUTHOR]

Ovarian cancer remains the disease with the highest associated mortality rate of gynecologic malignancy due to cancer metastasis. Rearrangement of actin cytoskeleton by cytoskeleton protein plays a critical role in tumor cell metastasis. MICAL-L2, a member of MICAL family, can interact with actin-binding proteins, regulate actin cross-linking and coordinate the assembly of adherens junctions and tight junctions. However, the roles of MICAL-L2 in tumors and diseases have not been explored. In this study, we found that MICAL-L2 protein is significantly up-regulated in ovarian cancer tissues along with FIGO stage and associated with histologic subgroups of ovarian cancer. Silencing of MICAL-L2 suppressed ovarian cancer cell proliferation, migration and invasion ability. Moreover, silencing of MICAL-L2 prevented nuclear translocation of β-catenin, inhibited canonical wnt/β-catenin signaling and induced the mesenchymal-epithelial transition (MET). Taken together, our data indicated that MICAL-L2 may be an important regulator of epithelial-mesenchymal transition (EMT) in ovarian cancer cells and a new therapeutic target for interventions against ovarian cancer invasion and metastasis. [ABSTRACT FROM AUTHOR]

24-h PM 2.5 samples were simultaneously collected at six sites in a subtropical city of South China during November–December, 2009. Particle-phase concentrations of polycyclic aromatic hydrocarbons (PAHs) and organic tracers such as hopanes for vehicular emissions (VE), levoglucosan for biomass burning (BB) and picene for coal combustion (CC) were determined. Meanwhile, their gas-phase concentrations were calculated from gas/particle (G/P) partitioning theory using the particle-phase concentrations. The 4 ring PAHs (fluoranthene to chrysene) had lower particle-phase fractions (10%–79%) than other species. Estimated BaP eq and lifetime cancer risk for particle-only (P-only) vs gas + particle (G + P) data sets showed similar values, indicating PAHs with 5–7 rings dominated the carcinogenicity of PAHs. Positive Matrix Factorization (PMF) was applied on both P-only and G + P data sets to estimate the source contributions to PAHs and their toxicity. Three common sources were identified: VE, BB and CC, with CC as the most significant source for both particulate (58%) and total (G + P, 40%) PAHs. While CC exhibited consistent contributions to BaP eq for P-only (66%) vs G + P (62%) solutions, VE and BB contributions were under- and overestimated by 68% and 47%, respectively by the P-only solution, as compared to the G + P solution. The results provide an insight on the impact of G/P partitioning on the source apportionment of PAHs and their toxicity. [ABSTRACT FROM AUTHOR]

Background & Aims: Monoamine oxidase A (MAOA), a catecholamine neurotransmitter degrading enzyme, is closely associated with neurological and psychiatric disorders. However, its role in cancer progression remains unknown. Methods: Hepatocellular carcinoma (HCC) tissue arrays (n=254) were used to investigate the correlation between MAOA expression and clinicopathological findings. In vitro invasion and anoikis assays, and in vivo intrahepatic and lung metastasis models were used to determine the role of MAOA in HCC metastasis. Quantitative real-time PCR, western blotting, immunohistochemical staining and HPLC analysis were performed to uncover the mechanism of MAOA in HCC. Results: We found that MAOA expression was significantly downregulated in 254 clinical HCC samples and was closely correlated with cancer vasoinvasion, metastasis, and poor prognoses. We then demonstrated that MAOA suppressed norepinephrine/epinephrine (NE/E)-induced HCC invasion and anoikis inhibition, and uncovered that the effects of NE/E on HCC behaviors were primarily mediated through alpha 1A (ADRA1A) and beta 2 adrenergic receptors (ADRB2). In addition to the canonical signaling pathway, which is mediated via adrenergic receptors (ADRs), we found that ADR-mediated EGFR transactivation was also involved in NE-induced HCC invasion and anoikis inhibition. Notably, we found that MAOA could synergize with EGFR inhibitors or ADR antagonists to abrogate NE-induced HCC behaviors. Conclusions: Taken together, the results of our study may provide insights into the application of MAOA as a novel predictor of clinical outcomes and indicate that increasing MAOA expression or enzyme activity may be a new approach that can be used for HCC treatment. [Copyright &y& Elsevier]

Abstract: Somatic cell nuclear transfer (SCNT) is a promising technique to produce mammalian transgenic clones. Only a small proportion of manipulated embryos, however, can develop into viable offspring. The abnormal growth and development of cloned animals, furthermore, are accompanied by aberrant lung development. Our objective was to investigate molecular background of lung developmental problems in transgenic (random insertion of exogenous DNA) cloned goats. We examined expression of 15 genes involved in growth regulation, imprinting, and epigenetic transcription in lung tissue of deceased transgenic cloned and normal goats of various ages. Compared with normal goats of the same age from conventional reproduction, expression of 13 genes (BMP4, FGF10, GHR, HGFR, PDGFR, RABP, VEGF, H19, CDKN I C, PCAF, MeCP 2 , HDAC1, and Dnmt3b) decreased in transgenic cloned goats that died at or shortly after birth; Expression of eight genes (FGF10, PDGFR, RABP, VEGF, PCAF, HDAC1, MeCP 2 , and Dnmt3b) decreased in fetal death of transgenic cloned goats. Expression of two epigenetic transcription genes (PCAF and Dnmt3b) decreased in disease death of transgenic cloned goats (1–4 months old). Disruptions in gene expression might be associated with the high neonatal mortality in transgenic cloned animals. These findings have implications in understanding the low efficiency of transgenic cloning. [Copyright &y& Elsevier]

Abstract: Myostatin, a member of the transforming growth factor-β family, acts as a negative regulator of skeletal muscle mass. In this study, myostatin-targeted caprine fibroblasts were obtained and subjected to SCNT to determine whether myostatin-knockout goats could be created. Fibroblasts from a 2-mo-old goat were transfected with a myostatin-targeted vector to prepare transgenic donor cells for nuclear transfer. After serum-starvation (for synchronization of the cell cycle), the percentage of transgenic fibroblasts in the G0/G1 phase increased (66.2% vs. 82.9%; P < 0.05) compared with that in the control group, whereas the apoptosis rate and mitochondrial membrane potential were unaffected (P > 0.05). There were no significant differences between in vivo- and in vitro-matured oocytes as recipient cytoplasts for rates of fusion (86.5% vs. 78.4%), pregnancy (21.6% vs. 16.7%), or kidding (2.7% vs. 0%). One female kid from an in vivo-matured oocyte was born, but died a few hours later. Microsatellite analysis and polymerase chain reaction identification confirmed that this kid was genetically identical to the donor cells. Based on Western blot analysis, myostatin of the cloned kid was not expressed compared with that of nontransgenic kids. In conclusion, SCNT using myostatin-targeted 2-mo-old goat fibroblasts as donors has potential as a method for producing myostatin-targeted goats. [Copyright &y& Elsevier]