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Identification of vulnerable sites defined by broadly neutralizing antibodies (bNAbs) on HIV-1 envelope (Env) is crucial for vaccine design, and we present here a vulnerable site defined by bNAb M4008_N1, which neutralizes about 40% of a tier-2 virus panel. A 3.2 Å resolution cryo-EM structure of M4008_N1 in complex with BG505 DS-SOSIP reveals a large, shallow protein epitope surface centered at the V3 crown of gp120 and surrounded by key glycans. M4008_N1 interacts with gp120 primarily through its hammerhead CDR H3 to form a β-sheet interaction with the V3 crown hairpin. This makes M4008_N1 compatible with the closed conformation of the prefusion Env trimer, and thus distinct from other known V3 crown mAbs. This mode of bNAb approaching the immunogenic V3 crown in the native Env trimer suggests a strategy for immunogen design targeting this site of vulnerability., Mapping of the HIV Env surface epitopes targeted by broadly neutralizing antibodies (bNAbs) is of great interest for HIV-1 vaccine design. Here, the authors present the 3.2 Å cryo-EM structure of the bNAb M4008_N1 in complex with BG505 DS-SOSIP, an engineered native-like Env trimer and observe that the bNAb epitope is centered at the V3 crown and that M4008_N1 uses its CDR H3 to form an extended β-sheet with the β-hairpin of the V3 crown in a conformation stabilized in the prefusion trimer.

Background Ten–eleven translocation methylcytosine dioxygenase 1 (TET1) is involved in DNA demethylation and transcriptional regulation, plays a key role in the maintenance of stem cell pluripotency, and is dysregulated in malignant cells. The identification of cancer stem cells (CSCs) driving tumor growth and metastasis is the primary objective of biomarker discovery in aggressive prostate cancer (PCa). In this context, we analyzed TET1 expression in PCa. Methods A large-scale immunohistochemical analysis of TET1 was performed in normal prostate (NOR) and PCa using conventional slides (50 PCa specimens) and tissue microarrays (669 NOR and 1371 PCa tissue cores from 371 PCa specimens). Western blotting, RT-qPCR, and 450 K methylation array analyses were performed on PCa cell lines. Genome-wide correlation, gene regulatory network, and functional genomics studies were performed using publicly available data sources and bioinformatics tools. Results In NOR, TET1 was exclusively expressed in normal cytokeratin 903 (CK903)–positive basal cells. In PCa, TET1 was frequently detected in alpha-methylacyl-CoA racemase (AMACR)–positive tumor cell clusters and was detectable at all tumor stages and Gleason scores. Pearson’s correlation analyses of PCa revealed 626 TET1-coactivated genes (r > 0.5) primarily encoding chromatin remodeling and mitotic factors. Moreover, signaling pathways regulating antiviral processes (62 zinc finger, ZNF, antiviral proteins) and the pluripotency of stem cells were activated. A significant proportion of detected genes exhibited TET1-correlated promoter hypomethylation. There were 161 genes encoding transcription factors (TFs), of which 133 were ZNF-TFs with promoter binding sites in TET1 and in the vast majority of TET1-coactivated genes. Conclusions TET1-expressing cells are an integral part of PCa and may represent CSCs with oncogenic potential.

AIM To discuss the effects and mechanism of microRNA-34a in cell apoptosis induced by osteoarthritis. METHODS Collection of the normal and osteoarthritis synovial tissues and measurements of the miRNA-34a and TGIF2 gene expression. In the cell experiment, the cells were divided into Control, Blank and miRNA inhibitor group. The cell proliferation and apoptosis of the different groups were measured by MTT and flow cytometry and the TGIF2 protein expression in the different groups was evaluated by WB assay. The correlation between TGIF2 and miRNA-34a was analyzed by Double luciferase experiment. RESULTS Compared with normal synovial tissues, the miRNA-34a gene expression was significantly up-regulated and TGIF2 gene expression was significantly suppressed in osteoarthritis synovial tissues (p < 0.001, respectively). The cell proliferation was significantly depressed and the cell apoptosis rate was significantly increased in miRNA inhibitor group compared with the Control group (p < 0.001, respectively). Using the WB assay it was shown that the TGIF2 protein expression of miRNA inhibitor group was significantly suppressed compared with that of Control group (p < 0.01). By Double luciferase assay, TGIF2 gene was one target gene of miRNA-34a. CONCLUSION miRNA-34a could induce osteoarthritis synovial cell apoptosis via regulation of TGIF2 in vitro (Fig. 6, Ref. 29).

We demonstrate applications of photonic-force optical coherence elastography (PF-OCE) on imaging three-dimensional (3D) micro-scale mechanical heterogeneity in collagen phantoms and cell-modified extracellular matrix. In collagen, we observed strong correlations between the measured mechanical heterogeneity and local OCT intensity in the vicinity of beads, which could be related to the presence of fibrous structures. Moreover, we quantitatively imaged micro-scale localized stiffening and increased heterogeneities in fibers surrounding an isolated NIH-3T3 fibroblast enclosed in a 2.5 mg/mL fibrin gel with a volumetric field-of-view (FOV) of ~80μm×350μm×50μm. Our results show that PF-OCE is able to characterize 3D micromechanical properties of biomaterials, and has the potential to observe dynamics of cell-ECM interactions.

AIM To evaluate the mixed-reality (MR) hologram, a novel technology based on two-dimensional images, which simulates three-dimensional (3D) images and provides a dynamic and interactive alternative, for its usefulness in the diagnosis and surgical planning of double outlet of the right ventricle (DORV). MATERIALS and METHODS Thirty-four patients who were suspected of DORV based on ultrasound findings underwent cardiac computed tomography angiography (CTA). The patients were assigned randomly to the MR holographic guidance (MRHG) group or the control group. For the patients in the MRHG group, the CTA images were converted into Standard Template Library (STL) files after segmentation, 3D reconstruction, colourisation, and transparentisation, and then exported for MR holographic visualisation. The CTA images of the patients in the control group were analysed using routine 3D reconstruction only. Diagnostic accuracy and surgical planning were compared between the two groups based on visualisation at surgery. RESULTS In the MRHG group, the 3D hologram observation was in concordance with the actual anatomical findings, and the DORV type was classified accurately in all patients. The diagnostic accuracy for the malformation was 95.5% in the MRHG group and 89.7% in the control group, but the difference was not significant (p=0.3). All the procedures were exactly the same as planned based on the 3D MR holographic model. The surgical planning time was shorter for the MRHG group (51.65 ± 11.11 min) than that for the control group (65.71 ± 18.07 min, p CONCLUSION MR 3D holograms may provide a clear and deeper anatomical perception of DORV and improve surgical planning.

Triple-negative breast cancer (TNBC) is an aggressive subtype characterized by extensive intratumoral heterogeneity. To investigate the underlying biology, we conducted single-cell RNA-sequencing (scRNA-seq) of >1500 cells from six primary TNBC. Here, we show that intercellular heterogeneity of gene expression programs within each tumor is variable and largely correlates with clonality of inferred genomic copy number changes, suggesting that genotype drives the gene expression phenotype of individual subpopulations. Clustering of gene expression profiles identified distinct subgroups of malignant cells shared by multiple tumors, including a single subpopulation associated with multiple signatures of treatment resistance and metastasis, and characterized functionally by activation of glycosphingolipid metabolism and associated innate immunity pathways. A novel signature defining this subpopulation predicts long-term outcomes for TNBC patients in a large cohort. Collectively, this analysis reveals the functional heterogeneity and its association with genomic evolution in TNBC, and uncovers unanticipated biological principles dictating poor outcomes in this disease., Triple-negative breast cancer is highly heterogeneous and aggressive. Here, the authors utilise single-cell RNA sequencing to investigate this heterogeneity, and discover a subpopulation of cells associated with metastasis and treatment resistance signatures, and linked to long term survival outcomes.

Synovial sarcoma (SyS) is an aggressive neoplasm driven by the SS18–SSX fusion, and is characterized by low T cell infiltration. Here, we studied the cancer–immune interplay in SyS using an integrative approach that combines single-cell RNA sequencing (scRNA-seq), spatial profiling and genetic and pharmacological perturbations. scRNA-seq of 16,872 cells from 12 human SyS tumors uncovered a malignant subpopulation that marks immune-deprived niches in situ and is predictive of poor clinical outcomes in two independent cohorts. Functional analyses revealed that this malignant cell state is controlled by the SS18–SSX fusion, is repressed by cytokines secreted by macrophages and T cells, and can be synergistically targeted with a combination of HDAC and CDK4/CDK6 inhibitors. This drug combination enhanced malignant-cell immunogenicity in SyS models, leading to induced T cell reactivity and T cell–mediated killing. Our study provides a blueprint for investigating heterogeneity in fusion-driven malignancies and demonstrates an interplay between immune evasion and oncogenic processes that can be co-targeted in SyS and potentially in other malignancies. Single-cell transcriptional profiling of primary human synovial sarcoma tumors suggests that combinatorial treatment with HDAC and CDK4/CDK6 inhibitors could enhance tumor immunogenicity.

Synovial sarcoma is an aggressive mesenchymal neoplasm, driven by the SS18-SSX fusion, and characterized by immunogenic antigens expression and exceptionally low T cell infiltration levels. To study the cancer-immune interplay in this disease, we profiled 16,872 cells from 12 human synovial sarcoma tumors using single-cell RNA-sequencing (scRNA-Seq). Synovial sarcoma manifests antitumor immunity, high cellular plasticity and a core oncogenic program, which is predictive of low immune levels and poor clinical outcomes. Using genetic and pharmacological perturbations, we demonstrate that the program is controlled by the SS18-SSX driver and repressed by cytokines secreted by macrophages and T cells in the tumor microenvironment. Network modeling predicted that SS18-SSX promotes the program through HDAC1 and CDK6. Indeed, the combination of HDAC and CDK4/6 inhibitors represses the program, induces immunogenic cell states, and selectively targets synovial sarcoma cells. Our study demonstrates that immune evasion, cellular plasticity, and cell cycle are co-regulated and can be co-targeted in synovial sarcoma and potentially in other malignancies.

In a Drosophila model of ALS, neuronal defects are associated with altered endosomal traffic of growth factor receptors and loss of growth-promoting signals. Manipulation of an endosomal recycling pathway suppresses these neuronal defects. The findings suggest that rerouting membrane traffic could be therapeutic in ALS., TAR DNA-binding protein 43 (TDP-43) is genetically and functionally linked to amyotrophic lateral sclerosis (ALS) and regulates transcription, splicing, and transport of thousands of RNA targets that function in diverse cellular pathways. In ALS, pathologically altered TDP-43 is believed to lead to disease by toxic gain-of-function effects on RNA metabolism, as well as by sequestering endogenous TDP-43 and causing its loss of function. However, it is unclear which of the numerous cellular processes disrupted downstream of TDP-43 dysfunction lead to neurodegeneration. Here we found that both loss and gain of function of TDP-43 in Drosophila cause a reduction of synaptic growth–promoting bone morphogenic protein (BMP) signaling at the neuromuscular junction (NMJ). Further, we observed a shift of BMP receptors from early to recycling endosomes and increased mobility of BMP receptor–containing compartments at the NMJ. Inhibition of the recycling endosome GTPase Rab11 partially rescued TDP-43–induced defects in BMP receptor dynamics and distribution and suppressed BMP signaling, synaptic growth, and larval crawling defects. Our results indicate that defects in receptor traffic lead to neuronal dysfunction downstream of TDP-43 misregulation and that rerouting receptor traffic may be a viable strategy for rescuing neurological impairment.

Moore et al. reports the first single-cell gene expression analysis in zebrafish blood to distinguish major blood lineages, identify new cell types, and delineate heterogeneity in T cell leukemia., Hematopoiesis culminates in the production of functionally heterogeneous blood cell types. In zebrafish, the lack of cell surface antibodies has compelled researchers to use fluorescent transgenic reporter lines to label specific blood cell fractions. However, these approaches are limited by the availability of transgenic lines and fluorescent protein combinations that can be distinguished. Here, we have transcriptionally profiled single hematopoietic cells from zebrafish to define erythroid, myeloid, B, and T cell lineages. We also used our approach to identify hematopoietic stem and progenitor cells and a novel NK-lysin 4+ cell type, representing a putative cytotoxic T/NK cell. Our platform also quantified hematopoietic defects in rag2E450fs mutant fish and showed that these fish have reduced T cells with a subsequent expansion of NK-lysin 4+ cells and myeloid cells. These data suggest compensatory regulation of the innate immune system in rag2E450fs mutant zebrafish. Finally, analysis of Myc-induced T cell acute lymphoblastic leukemia showed that cells are arrested at the CD4+/CD8+ cortical thymocyte stage and that a subset of leukemia cells inappropriately reexpress stem cell genes, including bmi1 and cmyb. In total, our experiments provide new tools and biological insights into single-cell heterogeneity found in zebrafish blood and leukemia.

OBJECTIVE The aim of this study was to evaluate the effects and mechanism of miRNA-31 in synovial cells apoptosis induced by RA. METHODS The miRNA-31 gene expressions were extracted from synovial tissues of normal and RA patients by RT-PCR and H et E staining. The synovial cells of RA patients were isolated and randomly divided into Control, Blank and miRNA groups. The cell apoptosis of difference groups were measured by flow cytometry; the TNF-α and IL-1β concentrations of difference groups were measured by Elisa assay; TLR4 and NF-κB proteins expressions were measured by WB assay and the correlation between TLR4 and miRNA-31 were evaluated by double luciferase target experiment. RESULTS The miRNA-31 gene expression was significantly suppressed in RA tissues (p

Background The levels of IL-33, a Th2 promoting cytokine, and the soluble form of its receptor ST2 were reported to be elevated in serum of patients with active systemic lupus erythematosus (SLE), suggesting a role of the IL-33/ST2 axis in the pathogenesis of SLE. Objectives This study aims to examine the effect of IL-33 in disease severity of murine lupus. Methods IL-33 was injected intraperitoneally 3 times per week to pre-diseased MRL/lpr mice aged 12 weeks for 6 weeks. Control group was given 1% BSA injection. Urine protein was monitored weekly by albustix and protein assay. Immunophenotyping of splenocytes was examined by flow cytometry. Splenic CD11b+monocytic cells were isolated by microbeads for mRNA examination. Results IL-33-treated mice (n=9) developed significantly less proteinuria compared to BSA-treated group (n=9). Kidney histology of the IL-33-treated group showed remarkably less mesangial deposit, diffuse proliferative glomerular changes and crescents, and had significantly lower renal composite score compared to controls (median 2.0 vs 9.9, p Conclusions Exogenous IL-33 led to significantly less proteinuria and renal inflammation. These mice had significantly higher splenic Treg cells with prominent Foxp3 expression. Isolated CD11b+cells from spleen and kidney extracts demonstrated mRNA levels of M2 macrophage polarisation. Disclosure of Interest None declared

Summary The aim of this study was to investigate the association between the single-nucleotide polymorphisms (SNPs) of the interleukin 22 (IL-22) gene and systemic lupus erythematosus (SLE) in a Chinese population. Three IL-22 SNPs (rs2227485, rs2227513 and rs2227491) were genotyped using SNaPshot SNP genotyping assays and identified by sequencing in 314 SLE patients and 411 healthy controls. The IL-22 level of serum was assessed by enzyme-linked immunosorbent assay (ELISA) kits. Data were analysed by spss version 17.0 software. We found that rs2227513 was associated with an increased risk of SLE [AG versus AA: adjusted odds ratio (aOR) = 2·24, 95% confidence interval (CI) = 1·22–4·12, P = 0·010; G versus· A: adjusted OR = 2·18, 95% CI = 1·20-3·97, P = 0·011]. Further analysis in patients with SLE showed that the AG genotype and G allele were associated with an increased risk of renal disorder in SLE (G versus A: aOR = 3·09, 95% CI = 1·30–7·33, P = 0·011; AG versus· AA: aOR = 3·25, 95% CI = 1·35–7·85, P = 0·009). In addition, the concentration of IL-22 was significantly lower in the rs2227513 AG genotype compared with AA genotype (P = 0·028). These results suggest that rs2227513 polymorphism might contribute to SLE susceptibility, probably by decreasing the expression of IL-22.

The cellular composition of H3K27M gliomas Diffuse midline gliomas with histone H3 lysine27-to-methionine mutations (H3K27M-glioma) are an aggressive type of childhood cancer with few options for treatment. Filbin et al. used a single-cell sequencing approach to study the oncogenic programs, genetics, and cellular hierarchies of H3K27M-glioma. Tumors were mainly composed of cells resembling oligodendrocyte precursor cells, whereas differentiated malignant cells were a smaller fraction. In comparison with other gliomas, these cancers had distinct oncogenic programs and stem cell–like profiles that contributed to their stable tumor-propagating potential. The analysis also identified a lineage-specific marker that may be useful in developing therapies. Science , this issue p. 331

Barrier-free facility is very important for handicapped daily life. Nevertheless, a good wheelchair design is helpfully supporting them. Unfortunately, sitting posture made handicapped perplexing, because the pressure ulcer problem occurs in their lower portion of body by traditional wheelchair. Since the major of the elders and the mobility patients are lower portion of body muscle or joint problems generated the travel inconvenience, therefore, it is necessary to design and study an innovative wheelchair that can aid the elders with mobility disability daily travelling a convenient transportation with multifunctional, easy-control method and superior user experience contributing to their lives. This investigation is going to discuss the possibility and feasibility of the smart wheelchair by Human Factors Engineering Technology to provide a comfortable and safety circumstance to the users, which can reduce the pressure ulcer problem by changing sitting posture with muscle signal control (EMG) and brain wave control (EEG).

In this paper, the influence of the depressant foenum-graecum (FGM) on the flotation of a nickel sulfide ore which contains hydrophobic talc and chlorite was systematically investigated through single mineral flotation, bench-scale and industrial-scale flotation tests. The flotation results indicated that FGM had excellent depression selectivity for talc and chlorite minerals and little depression effect on copper–nickel minerals. Compared with the conventional depressant carboxymethyl cellulose, the FGM scheme increased the recovery of copper in Cu–Ni concentrates over 25% and nickel in Cu–Ni concentrates over 16% as well as simplified the structure of flotation process. In addition, the adsorption mechanisms of FGM on talc and copper–nickel minerals were studied by adsorption, zeta-potential and FTIR measurements, which further demonstrated that the adsorption of FGM on talc was much stronger than that on pentlandite, FGM could selectively adsorb on talc surfaces through chemical adsorption while a weak hydrogen bond or electrostatic attraction occurred between FGM and copper–nickel minerals.

A colored phenotype is an important feature of ornamental fish. In mammals, microphthalmia-associated transcription factor (MITF) was found to regulate the development of melanocytes. In this study, the mitfa cDNA was first cloned from the Japanese ornamental (Koi) carp (Cyprinus carpio L.), an important ornamental freshwater fish. The full-length cDNA of the mitfa gene contains 1634 bp, coding for 412 amino acids in Koi. The identity degree of mitfa amino acid sequences between the Koi carp and zebrafish is 92.9%. We tested the expression of the mitfa gene in several varieties of Koi using reverse transcription-polymerase chain reaction and found that the mitfa gene is highly expressed in the skin tissues of the Taisho sanke and the Procypris merus. Interestingly, the mitfa gene was also expressed in the Kohaku and Yamabaki ogon, although melanocytes were not observed in the skin. Koi carp embryos were transparent and colorless, while after hatching, different types of pigment cells successively emerged in a fixed order. In Taisho sanke, melanocytes first appeared in the trunk at approximately 12 days of age. Subsequently, there was a large area of melanocytes by 30 days of age. The expression level of the mitfa mRNA was low in early embryos and newly hatched larvae, and increased to high levels in 30-day-old fry. The results show that the mitfa gene is involved in regulating fish body color in the development of both melanocytes and pigment cells.

We investigated the effects of smoking and aging on proteins involved in the forkhead box O3 (FOXO3) signaling pathways in the lungs. Sixteen senescence-accelerated mouse-resistant 1 (SAMR1) and senescence-accelerated mouse-prone 8 (SAMP8) mice at 3 months of age were divided into a normally aged, smoke-exposed group (4 SAMR1 mice), a normally aged, air-exposed group (4 SAMR1 mice), an aging-accelerated, smoke-exposed group (4 SAMP8 mice), and an aging-accelerated, air-exposed group (4 SAMP8 mice). Expression of genes and proteins related to the FOXO3 signaling pathways in each group was examined by western blot analysis and immunohistochemistry. FOXO3a expression was significantly increased in the normally aged, air-exposed group compared with the aging-accelerated, air-exposed group. FOXO3a expression was significantly reduced in the aging-accelerated, smoke-exposed group compared with the aging-accelerated, air-exposed group. Sirtuin 1, manganese superoxide dis-mutase, and phosphatidylinositol 3-kinase (PI3K)/Akt expression decreased significantly in the smoke-exposed groups compared with the air-exposed groups and in the aging-accelerated groups compared with the normally aged groups. Signal transduction pathways mediated by the transcription factor FOXO3a (such as the PI3K/Akt pathway) may be involved in the accelerated aging of lung tissues in chronic obstructive pulmonary disease. Smoking inactivates the transcription factor FOXO3, thus accelerating lung tissue aging during chronic obstructive pulmonary disease.

Tumor subclasses differ according to the genotypes and phenotypes of malignant cells as well as the composition of the tumor microenvironment (TME).We dissected these influences in isocitrate dehydrogenase (IDH)-mutant gliomas by combining 14,226 single-cell RNA sequencing (RNA-seq) profiles from 16 patient samples with bulk RNA-seq profiles from 165 patient samples. Differences in bulk profiles between IDH-mutant astrocytoma and oligodendroglioma can be primarily explained by distinct TME and signature genetic events, whereas both tumor types share similar developmental hierarchies and lineages of glial differentiation. As tumor grade increases, we find enhanced proliferation of malignant cells, larger pools of undifferentiated glioma cells, and an increase in macrophage over microglia expression programs in TME. Our work provides a unifying model for IDH-mutant gliomas and a general framework for dissecting the differences among human tumor subclasses., National Cancer Institute (U.S.) (Grant P30-CA14051)

Ischemic stroke can lead to loss of neurologic functions. It occurs due to obstruction in blood supply to the brain. It has been proposed that C807T(C/T) polymorphism within the platelet glycoprotein gene may be associated with density and function of glycoprotein Ia/IIa receptors and contributes to the pathogenesis of thrombotic disease. We assessed the association between C807T(C/T) and risk of ischemic stroke. Databases such as PubMed, Medline, Springer, Elsevier Science Direct, Cochrane Library, Google scholar, Wanfang Data (Chinese), and Chinese National Knowledge Infrastructure (CNKI, Chinese) were used to search for relevant studies. We found 16 eligible studies, which totaled to 4897 (case group 2340; control group 2557) participants. Overall, our results showed significant associations between C807T(C/T) polymorphism and risk of ischemic stroke based on T-allele comparisons (T vs C, pooled OR = 0.78, 95%CI = 0.68-0.90, P0.01), TT vs CC comparisons (pooled OR = 0.58, 95%CI = 0.42-0.81, P0.01), recessive models (TT vs TC + CC, pooled OR = 0.72, 95%CI = 0.59-0.87, P0.01) and dominant models (TT + TC vs CC, pooled OR = 0.70, 95%CI = 0.54-0.92, P0.05). There was no association in TC vs CC comparisons (pooled OR = 0.81, 95%CI = 0.63-1.04, P0.05). Subgroup analyses stratified according to Hardy-Weinberg equilibrium, sample size, and ethnicity also demonstrated significant associations between the two variables. Therefore, C807T(C/T) polymorphism in the platelet glycoprotein gene may be associated with susceptibility to ischemic stroke, and the T allele at this locus may decrease risk to ischemic stroke.

The V1V2 region of the HIV-1 envelope is the target of several broadly neutralizing antibodies (bNAbs). Antibodies to V1V2 elicited in the RV144 clinical trial correlated with a reduced risk of HIV infection, but these antibodies were without broad neutralizing activity. Antibodies targeting V1V2 also correlated with a reduced viral load in immunized macaques challenged with simian immunodeficiency virus (SIV) or simian/human immunodeficiency virus (SHIV). To focus immune responses on V1V2, we engrafted the native, glycosylated V1V2 domain onto five different multimeric scaffold proteins and conducted comparative immunogenicity studies in macaques. Vaccinated macaques developed high titers of plasma and mucosal antibodies that targeted structurally distinct V1V2 epitopes. Plasma antibodies displayed limited neutralizing activity but were functionally active for ADCC and phagocytosis, which was detectable 1–2 years after immunizations ended. This study demonstrates that multivalent, glycosylated V1V2-scaffold protein immunogens focus the antibody response on V1V2 and are differentially effective at inducing poly-functional antibodies with characteristics associated with protection.

SUMMARY When hens are inseminated by different types of roosters, natural elimination of the influence of insemination by a previous rooster may require some time, reducing the breeding activity of the hens. Two separate experiments were conducted to determine the effects of sperm competition between 2 breeder males on reducing the time interval which eliminated the influence of previous artificial insemination (AI). At the conclusion of the experiments, sperm retained their activity even after 20 d in a noncompetitive situation. When females were artificially inseminated by different types of roosters, if the time interval between AI was less than 12 d, 14 d were required to eliminate the influence of the previous AI in the sperm competition even if the females were repeatedly inseminated by the second type of breeder males. However, if the duration between AI was more than 12 d, AI once by a different type of breeder males was sufficient to prevent the spermatization of the previous male. In summary, the current study suggests that maintaining a time interval of 12 d between inseminations and repeatedly inseminating females by a different type of breeder male every 4 d allowed the elimination of the influence of the previous AI within 14 d. This duration was 7 d less than that required for the natural elimination of the influence of the previous AI, and such a situation could be easily manipulated.

Background: There are well established disparities, related to ethnicity, in the presentation, distribution of subtypes and prognosis of breast cancer. To understand the possible biological bases for these differences, it is important to determine in an unbiased manner the ethnicity/geographical origin of patients presenting with breast cancer. We have downloaded Affymetrix SNP 6.0 data generated by The Cancer Genome Atlas project (TCGA) and have extracted ethnically informative SNPs to infer the ethnicity/geographical origin of these patients. Materials and Methods: The level one SNP data (Affymetrix SNP Array 6.0) for 536 normal DNAs from IBC patients as well as other relevant data were downloaded from the TCGA project data portal (https://tcga-data.nci.nih.gov/tcga/tcgaHome2.jsp). 103 samples were excluded for technical reasons (lack of subtype data or technical outliers). Bioconductor package “crlmm” was used for genotype calling. The HapMap PCA loadings for Affymetrix SNP probes were downloaded from http://www.stats.ox.ac.uk/~davison/software/shellfish/shellfish.php. Probe annotations were extracted from the Affymetrix GenomeWideSNP_6_na30.annot.csv file. “EIGINSTRAT” was used for Principle Component Analysis (PCA). Samples with unknown ethnicity were manually assigned based on their HapMap classification. Results: Samples were clustered and ethnicity inferred for three major ethnic groups (White, African and Asian) based on 168,905 informative SNPs. These clusters were generally in agreement with the clinical assignment of White, Black or African American and Asian. 10 samples, clinically assigned as Black or Africa American, were placed between the White and African clusters based on the HapMap clustering suggesting admixture. This contrasts other large breast cancer cohorts such as Curtis, et al. 2012, where there were virtually no African genotype patients and there was evident admixture on the European – Asian axis. 113 samples with unknown ethnicity were assigned based on their HapMap classification. Finally there are 371 Whites, 25 African, 20 Asians, and 10 Admixtures with subtype information available. Subtype distribution were LumA 176 (46.68%), LumB 90 (23.87%), Her2 37 (9.81%), Basal 68 (18.04%) in White group; LumA 15 (33.33%), LumB 8 (17.78%), Her2 9 (20%), Basal 13 (28.89%) in non-Whites. The subtype distribution between White and non-White populations was significantly different (p < 0.05, chi-square test), 7 Normal-like samples were excluded due to the low count. The Basal subtype had higher frequency among Africans compared to other ethnic groups. The LumB subtype also showed higher frequency in the 10 admixture samples, half of which were of the LumB subtype. Discussion: Inferring ethnicity/geographical origin by genotype removes bias known to occur in self reporting. We have successfully classified the TCGA breast cancer cohort into different ethnicity groups based on their ethnically informative SNP data. Although the SNP-based classification is generally in agreement with the clinical assignment, some samples are identified as admixtures between White and African. The subtype distribution varies by ethnicity/geographical origin and in the admixed population. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-09-03.

Polycomb repressive complex (PRC2) contains several proteins, including embryonic ectoderm development (EED), suppressor of zeste 12 (SUZ12) and enhancer of zeste homolog 2 (EZH2). Excessive EZH2 concentrations have been reported as a marker of aggressive breast cancer and associated with invasion and cancer progression. EZH2 levels were elevated in patients with invasive breast carcinoma relative to normal or atypical hyperplasia. Except for the C-terminal SET domain, which functions as a histone-lysine N-methyltransferases, EZH2 has an N-terminal alpha helix region, which forms tight complex with EED protein. The in vivo enzymatic activity of EZH2 relies on and be tightly regulated by the interaction with EED. We designed series of artificial peptides trying to block the interaction between EZH2 and EED. By using label-free surface plasmon resonance (SPR) technology, the dynamic binding capacities of these peptides were tested. There were 3 leading structures standing out from the screening of more 80 peptides. After several round of co-crystal structure based optimization, the binding capacity of one inhibitor was reached to several nM levels, which indicated the availability for being a drug candidate. Finally, the biological activities of the inhibitory peptides were tested in several breast cancer cell lines. The PRC2 inhibitory peptides in this study are the first time reported PRC2 targeting epigenetic molecules with biological activity in breast cancer. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P2-09-06.

Background: Given the lack of effective targeted therapies for triple-negative (TN) breast cancer patients, a better understanding of the mechanisms of growth and invasion of these tumors will provide valuable insight into developing novel approaches to lower the mortality associated with TN breast cancer. There is substantial evidence indicating that the canonical Wnt signaling pathway is activated in TN breast cancer tissues. Previous studies report that FH535, a small molecule inhibitor of the Wnt signaling pathway, decreases growth of cancer cells but not normal fibroblast cells, suggesting this pathway plays a role in tumor progression and metastasis. In this study, we test FH535 as a potential inhibitor of migration, invasion, and growth against malignant phenotypes of breast cancer. Methods and Results: We show that FH535 significantly inhibits growth, migration, and invasion of TN breast cancer cells (MDA-MB-231 and HCC38) using established protocols, implicating the Wnt signaling pathway plays a role in tumor progression and metastasis. FH535 is a potent growth inhibitor for TN breast cancer cells (MDA-MB-231 and HCC38), but not in other breast cancer cells (MCF-7, T-47D, or SK-BR-3) when cultured in three dimensional type I collagen gel. Additionally, western blotting analysis shows that treatment of MDA-MB-231 cells with FH535 markedly inhibits the expression of NEDD9, but not FAK, Src, or p130Cas, suggesting specificity of the Wnt signaling pathway in regulating gene expressions. Co-immunoprecipitation studies suggest that NEDD9 is specifically immunoprecipitated with CSPG4, but not with β1 integrin or CD44, implying CSPG4 and NEDD9 form a molecular complex for facilitating tumor migration, invasion, and growth. Statistical analysis (ANOVA) of The Cancer Genome Atlas (TCGA) project gene expression data (https://tcga-data.nci.nih.gov/) suggests that tumors from patients with basal-type breast cancer had significantly higher CSPG4 expression than tumors from patients with other subtypes, with all the p values Discussion: In this study, we demonstrate the pivotal role of the canonical Wnt signaling pathway in enhancing tumor migration, invasion, and growth of TN breast cancer cells MDA-MB-231 and HCC38. We identified NEDD9 as a potential downstream target of the Wnt signaling pathway and found an association between NEDD9 and CSPG4 in MDA-MB-231 cells. Our results suggest a novel mechanism of migration, invasion, and growth of tumor cells involving a molecular complex between CSPG4 and NEDD9, which could be drug targets in TN and possibly Luminal A breast cancer patients. More comprehensive characterization of canonical Wnt signaling pathway by genomic and biological studies of breast cancer cells are clearly required for generating better treatments. The views expressed in this article are those of the author and do not reflect the official policy of the Department of Defense, or U.S. Government. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P1-05-09.

Background: Invasive breast cancer (IBC) has been classified into four major subtypes based on gene expression profiling. The luminal A subtype (LA) has the best prognosis, when compared to luminal B (LB), HER2+, and basal-like (Basal). Ki67 by gene expression or immunohistochemistry (IHC) is commonly used as a proliferation index. The function of Ki67 in proliferation remains unknown. AURKA (STK15) is known to play an important role in mitosis, and is a component of the 21-gene recurrence score of the Oncotype Dx. With multiple platforms of molecular data available from hundreds of IBC tissues in The Cancer Genome Atlas project (TCGA), we sought to study the association of AURKA with different IBC subtypes and explore its use as a proliferation marker in IBCs. Methods: Gene expression (Agilent, log2 transformed), relative DNA copy number (CN, Affymetrix SNP 6.0), and exome sequence mutation (Illumina) data for 459 IBC cases were downloaded from the TCGA data portal. PAM50 classification results of all samples were obtained from the TCGA breast cancer AWG group and included 203 LA, 113 LB, 51 HER2+, 84 Basal-like, and 8 Normal-like which were not used in this study due to the low numbers. Kruskal-Wallis tests were used to evaluate the differences among four subtypes on AURKA expression and CN, followed by Wilcoxon Mann-Whitney test with Bonferroni adjustment for pairwise analyses. Pearson's Correlation Coefficient was used for correlation analyses. All statistical analyses were performed using SAS and R, and two-sided, p values Results: There was a significant difference among IBC subtypes, in gene expression as well as in CN (p values < 0.0001). AURKA mRNA levels were significantly lower in LA (mean±SD, −2.61±0.63) compared to LB (−1.45±0.78), HER2+ (−1.38±0.61), and Basal (−1.26±0.62) subtypes (p values all < 0.0001). No significant difference was detected between other subtype pairs. In CN analysis, Basal (0.09±0.22) was lower than HER2+ (0.32±0.308, p < 0.0002) and LB (0.33±0.41, p < 0.0001), and LA (0.14±0.28) is lower than HER2 (p < 0.0016) and LB (p < 0.0001), but no other significant CN difference between the subtypes were found. The means and SDs are provided for reference only. No correlation of p53 mutation status and AURKA expression were observed. However, AURKA gene expression level is correlated with MKI67 gene expression (R = 0.69, p < 2.2e−16), and its correlation with PAM50 proliferation score is even higher (R = 0.80, p < 2.2e−16). Discussion: Using the TCGA data we observed that the mean gene expression level of AURKA is significantly lower in LA than the other IBC subtypes, by more than 50% (note the log2 transformation). This differential expression is not completely due to CN changes (especially for the Basal subtype). There is a strong association with other tumor cell proliferation markers such as the MKI67 gene and the PAM50 proliferation score. We are using computational and laboratorial studies to better understand the role of AURKA in the etiology of invasive breast cancers. The views expressed in this article are those of the author and do not reflect the official policy of the Department of Defense, or U.S. Government. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P2-05-21.

To investigate the use of second-generation dual-source high-pitch computed tomography in obtaining confident diagnostic image quality using a low radiation dose in young patients with congenital heart disease (CHD).From July 2014 to June 2016, 50 consecutive children4 years with complex CHD underwent electrocardiography (ECG)-triggered dual-source computed tomography (CT). The patients were assigned randomly to two groups: high-pitch (pitch 3.4) spiral dual-source CT acquisition (group A) and retrospectively spiral dual-source CT acquisition (group B). The image quality, diagnostic accuracy, coronary artery origin, course demonstration, and radiation exposure were compared between the two groups.Fifty examinations were performed (group A, 25; group B, 25). There were no significant differences in image quality, diagnostic accuracy, coronary artery origin, and course demonstration between the two groups. The image quality scores were 1.3±0.4 in group A and 1.1±0.3 in group B (p=0.2). The diagnostic accuracy was 100% in both groups. The coronary arteries were traceable in 80% in group A and 84% in group B (p=0.7). A single coronary artery was identified in one case in group A and the left anterior descending (LAD) branch originated from the right coronary artery (RCA) in one case in group B. There were significant differences in the effective doses between the two groups (0.40±0.20 mSv in group A and 2.7±1.0 mSv in group B, p0.05).Intra-cardiac and extra-cardiac malformation, coronary artery origin, and course malformation can be visualised clearly using a high-pitch ECG-triggered dual-source CT with a low radiation dose and good image quality in patients with CHD.

BackgroundThe present study investigated alteration of brain resting-state activity induced by antidepressant treatment and attempted to investigate whether treatment efficacy can be predicted at an early stage of pharmacological treatment.MethodForty-eight first-episode medication-free patients diagnosed with major depression received treatment with escitalopram. Resting-state functional magnetic resonance imaging was administered prior to treatment, 5 h after the first dose, during the course of pharmacological treatment (week 4) and at endpoint (week 8). Resting-state activity was evaluated in the course of the 8-week treatment and in relation to clinical improvement.ResultsEscitalopram dynamically modified resting-state activity in depression during the treatment. After 5 h the antidepressant induced a significant decrease in the signal in the occipital cortex and an increase in the dorsolateral and dorsomedial prefrontal cortices and middle cingulate cortex. Furthermore, while remitters demonstrated more obvious changes following treatment, these were more modest in non-responders suggesting possible tonic and dynamic differences in the serotonergic system. Changes after 5 h in the caudate, occipital and temporal cortices were the best predictor of clinical remission at endpoint.ConclusionsThis study revealed the possibility of using the measurement of resting-state neural changes a few hours after acute administration of antidepressant to identify individuals likely to remit after a few weeks of treatment.

This paper proposes a robotics therapeutic massage evaluation system using electroencephalogram (EEG) signal analysis approach. An anthropomorphic dual arm robot is developed in this work for massage application. Compare to other massage systems, our dual arm robot can provides diversified massage techniques through the impedance control of both Cartesian space and joint space. In order to evaluate the effectiveness of robotic therapeutic massage, the experiments are conducted by recording EEG signals of subject before and after robotic massage. Independent Components Analysis (ICA) is used to filter out artifacts of EEG signals. After signal processing, the power of delta, alpha and beta rhythms are analyzed. The experimental results show an increase in delta power and a decrease in alpha power which represents the relaxation response of subjects. These results give the scientific proof of the effectiveness of robotic therapeutic massage.

Inspired by the human torso, humanoid robots usually equipped with active waist joints. For arbitrary walking patterns or whole body motions, the well-designed waist trajectories can greatly enhance the motion of robot legs and allow the robot to perform larger walking step length or crossing higher stair gaps. In this paper, we develop an effective online algorithm for generating waist and arm trajectories for humanoid robots having a waist joint. The waist trajectory generation problem is formulated into a Quadratic Programming problem. The Receding Horizon Control technique is used to make the algorithm relative faster to integrate it into online control systems and have the ability to confront perturbations. The arm swinging motion is then generated to eliminate the angular momentum generated by the swinging leg and the waist motion. Finally, we apply our algorithm on RENBO, which is the humanoid robot developed in our lab. The experimental result and analysis demonstrate that the algorithm can generate continuous and smooth waist and arm trajectory, making the resulting leg motion much smoother.

Although human tumours are shaped by the genetic evolution of cancer cells, evidence also suggests that they display hierarchies related to developmental pathways and epigenetic programs in which cancer stem cells (CSCs) can drive tumour growth and give rise to differentiated progeny1. Yet, unbiased evidence for CSCs in solid human malignancies remains elusive. Here we profile 4,347 single cells from six IDH1 or IDH2 mutant human oligodendrogliomas by RNA sequencing (RNA-seq) and reconstruct their developmental programs from genome-wide expression signatures. We infer that most cancer cells are differentiated along two specialized glial programs, whereas a rare subpopulation of cells is undifferentiated and associated with a neural stem cell expression program. Cells with expression signatures for proliferation are highly enriched in this rare subpopulation, consistent with a model in which CSCs are primarily responsible for fuelling the growth of oligodendroglioma in humans. Analysis of copy number variation (CNV) shows that distinct CNV sub-clones within tumours display similar cellular hierarchies, suggesting that the architecture of oligodendroglioma is primarily dictated by developmental programs. Subclonal point mutation analysis supports a similar model, although a full phylogenetic tree would be required to definitively determine the effect of genetic evolution on the inferred hierarchies. Our single-cell analyses provide insight into the cellular architecture of oligodendrogliomas at single-cell resolution and support the cancer stem cell model, with substantial implications for disease management.

Robot-assisted minimally invasive surgery (RMIS) has become widespread because of its accuracy, stability and intuition. In the traditional laparoscopic surgery, an assistant is essential to help the surgeon in controlling the laparoscope. However, surgeons need to communicate with the assistant, and it is also difficult for an assistant manipulate the laparoscope to an anatomical position that the surgeon desired. Accordingly, the objective of this paper is to develop the Intuitively Robotic Laparoscope System (IRLS) with a more intuitive means to conquer these problems. With the gyroscope mounted on the head, the surgeon can dominate the laparoscope maneuver directly. To maneuver multi degree of freedoms and increase safety, a pedal is used as a power and mode switch. Furthermore, an algorithm is also adopted to adjust the angle collected by the gyroscope to make this system more ergonomic. Finally, an experimental result demonstrates that the data from the gyroscope being used is feasible and the error is acceptable.

Objectives To describe the various factors that come into play when a breast-feeding mother is taking medications, including use of prescription drugs, over-the-counter medications, recreational drugs, galactogogues, and herbal remedies and to provide a framework used for counseling breast-feeding women. Setting Community and hospital pharmacy and health care settings. Practice description Consultative services provided to breast-feeding mothers who had been prescribed or were using medications. Main outcome measures Use of pharmacokinetic factors, maternal and child factors, a list of questions to ask breast-feeding mothers, and a stepwise approach to counsel breast-feeding mothers on the compatibility of using medications while breast-feeding. Results By positive intervention of pharmacists and health care providers, up to 1 million breast-feeding mothers, who must use medications, can continue to breast-feed while taking medications. Conclusion Objectively weighing the benefits of drugs and breast-feeding versus the risks of drugs and not breast-feeding, in most cases, allows for pharmacists to give current and practical advice to mothers and other health professionals who counsel mothers.

Polished specimens of AA2024-T3 alloy were immersed for up to 120 min in 0.1 M NaCl. The development of corrosion was monitored using scanning electron microscopy with energy dispersive X-ray spectroscopy (EDXS) and particle induced X-ray emission spectroscopy (PIXE). Both techniques revealed the intermetallic (IM) particle distributions and attack sites as distinguished by detection of chloride species. The earliest stages of attack involved localized attack around isolated IM particles as reported in Part I. Additionally attack occurred on a larger scale developing rapidly with rings of corrosion product surrounding clusters of IM particles. There were significantly higher numbers of IM particles within the corrosion rings, indicating that local clustering played an important role in co-operative corrosion.

This journal suppl. entitled: Abstracts of the 26th Annual Meeting of the European Society of Human Reproduction and Embryology, Rome, Italy, 27-30 June 2010

Studies investigating the association between angiotensin-converting enzyme (ACE) insertion/deletion polymorphisms and the risk of acute respiratory distress syndrome (ARDS) report conflicting results. The objective of the present study was to summarize quantitatively the evidence for such a relationship. Two investigators independently searched the MEDLINE (January 1966 - October 2009) and EMBASE (January 1980 - October 2009) databases. This meta-analysis included six case-control studies, which included 514 ARDS cases and 2619 controls. The combined results based on all studies showed that patients with ARDS had a significantly higher frequency of the ACE deletion/deletion (DD) genotype (odds ratio [OR] 1.56; 95% confidence interval [CI] 1.22, 2.00) than controls. When stratifying for race, no significant differences in genotype distribution were found except that Caucasian patients with ARDS had a significantly higher frequency of the DD genotype (OR 1.65; 95% CI 1.27, 2.13) than controls. This meta-analysis suggests that ACE insertion/deletion polymorphisms may be associated with ARDS among Caucasians.

The corrosion of aluminium alloy 2024-T3 (AA2024-T3) was studied as a function of immersion time from 2.5 to 120 min in 0.1 M aqueous NaCl solution. At immersion times as short as 5 min, rings of corrosion product of 100 to 200 μm diameter, containing smaller domes of corrosion product, were observed using SEM. The domes of corrosion product had greater chloride concentrations than elsewhere on the surface and represented sites of anodic attack. As the immersion time was increased, significant grain boundary attack was observed within the rings of corrosion product. Analyses of Particle Induced X-ray Emission (PIXE) maps of the corroded surfaces showed a significantly higher number of IM particles around the chloride attack sites than the average particle density for the maps, indicating clustering of IM particles. These results suggest a co-operative corrosion effect as a result of clustering of the IM particles. A mechanism for the generation of the corrosion rings is discussed.

Although the expression of heparanase is associated with invasion and metastasis of various human cancers, the effects of heparanase on human osteosarcoma have not been evaluated. We showed that down-regulating the expression of heparanase significantly reduced proliferation and invasion of human U-2OS osteosarcoma cells. Furthermore, heparanase silencing by short-hairpin RNA (shRNA) was associated with decreased hypoxia-inducible factor-1α (HIF-1α) level, implying that heparanase was associated with the expression of HIF-1α. This result was confirmed by immunohistochemistry analysis. In osteosarcoma tissues, immunohistochemical results revealed that heparanase expression had a close correlation with that of HIF-1α and they had a strong relation with presence of pulmonary metastasis (P

Objective: Low vitamin D (25 OH vitamin D) is implicated in the development of diabetes and the metabolic syndrome. We examined whether hypovitaminosis D has a clinically significant impact on glycaemia, metabolic status and inflammatory markers in Chinese patients with established type 2 diabetes. Methods: Characteristics of 109 patients aged over 50 years were stratified by 25 OH vitamin D status. Patients identified as 25 OH vitamin D deficient (≤ 50 nmol/L) received cholecalciferol 2,000 IU daily for three months. Measurement of HbA1c, metabolic syndrome parameters, 25 OH vitamin D, calcium, phosphate, PTH, hsCRP and ferritin were taken at baseline and then 25 OH vitamin D, PTH, calcium, phosphate monthly for three months in those on replacement therapy. Results: Vitamin D deficiency was common, affecting 36% of patients. There was no impact of hypovitaminosis D on metabolic syndrome status, HbA1c or insulin use (p ≥ 0.4 for all) and no association between 25OHVitD and ferritin or hsCRP (p ≥ 0.3 for all). Neither BMI nor the metabolic syndrome affected the incremental rise in 25OHVitD levels during supplementation. Conclusion: There is no relationship between hypovitaminosis D and metabolic control or inflammatory markers in established type 2 diabetes.This suggests that at least in Chinese populations, the effect of low vitamin D is not clinically significant once diabetes is established. Future 25OHVitD intervention trials should therefore focus on prevention in pre-diabetes.

OBJECTIVE: This systematic review assesses the effectiveness of ventral rectopexy (VR) surgery for treatment of rectal prolapse (RP) and rectal intussusception (RI) in adults. Method MEDLINE, EMBASE, Scopus and other relevant databases were searched to identify studies. Randomized controlled trials or nonrandomized studies with more than 10 patients receiving ventral mesh rectopexy surgery were considered for the review. RESULTS: Twelve nonrandomized case series studies with 728 patients in total are included in the review. Seven studies used the Orr-Loygue procedure (VR with posterior rectal mobilization to the pelvic floor) and five studies used VR without posterior rectal mobilization. Overall weighted mean percentage decrease in faecal incontinence (FI) rate was 45%. The weighted mean percentage decrease in constipation rate was 24%. Weighted mean recurrence rate was 3.4%. CONCLUSIONS: There are limitations in published literature on VR. The available data indicate that VR has low recurrence and improves FI in patients suffering from these conditions. There is a greater reduction in postoperative constipation if VR is used without posterior rectal mobilization.