本文以漯河地区四处龙山时代晚期中小型遗址(寺疙瘩、凌云台、善德、大悲寺)调查发现的炭化植物遗存为切入点,探讨淮河上游南部地区的农业结构.浮选出的炭化植物遗存包括粟、黍、水稻、大豆属、小麦5种农作物,狗尾草属、马唐属等旱地与荒地杂草.量化分析显示漯河地区龙山时代晚期中小型遗址的作物结构以粟为主,黍为辅,普遍伴出少量稻、大豆,可能出现小麦.综合已公布的研究资料,认为龙山时代晚期淮河上游广大区域内的农业结构趋于一致.

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Having a functional bioassay that is MOA-based, accurate, precise, robust and reproducible is critical for the development of antibody-based biologics. We have developed reporter bioassays that meet these criteria for a broad range of antibody modalities including Fc effector function, immune checkpoint modulation, bispecific antibody engagement, cytokine modulation, and others. Here we will present the latest technology advancements and bridging of the PBMC based ADCC cell killing assay with reporter based ADCC Bioassay. Citation Format: Mei Cong, Pete Stecha, Aileen paguio, brock binkowski, Zhi-Jie Cheng, Frank Fan. Bridging MOA-based reporter bioassays with PBMC based ADCC for immunotherapy drug development [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2393.

Abstract Immunomodulatory biologics, which elicit their therapeutic effects by inducing, inhibiting, or modulating immune responses, have shown therapeutic utility in a variety of clinical conditions including cancer and autoimmune disease. As a result, biologics therapeutics, and monoclonal antibodies (mAb) in particular, are a dominant and increasing focus of pharmaceutical pipelines. However, due to their structural and biological complexity, mAb biologics and biosimilars require unique functional and analytical tools for development and characterization. We have developed a suite of cell and reporter-based bioassays to advance the development and commercialization of mAb biologics. Specifically, we have developed cell-based bioassays in “thaw-and-use” format for the following Immune Checkpoint and Immunostimulatory receptor targets: PD-1/PD-L1, CTLA-4, LAG-3GITR, 4-1BB, CD40, OX40T Cell Activation These reporter-based bioassays offer high specificity, sensitivity, and reproducibility, and demonstrate the performance required for use in antibody screening, potency determination, and stability studies.

Studies have shown that people are better at recognizing human faces from their own-race than from other-races, an effect often termed the Own-Race Advantage. The current study investigates whether there is an Own-Race Advantage in attention and its neural correlates. Participants were asked to search for a human face among animal faces. Experiment 1 showed a classic Own-Race Advantage in response time both for Chinese and Black South African participants. Using event-related potentials (ERPs), Experiment 2 showed a similar Own-Race Advantage in response time for both upright faces and inverted faces. Moreover, the latency of N2pc for own-race faces was earlier than that for other-race faces. These results suggested that own-race faces capture attention more efficiently than other-race faces.

Recent research has observed two basic eye movement patterns in face recognition: central (fixations are around the nose) and triangular (fixations are on the two eyes and mouth). Here we aim to examine the link between eye movement patterns and engagement of global/local attention. Eye movements of Asian participants were monitored in an old/new face judgment task. Participants received global or local priming by matching two Navon stimuli at the global or local level respectively between the study and recognition phase, and in the beginning of each recognition trial. A hidden Markov model (HMM) based approach (Chuk, Chan, & Hsiao, 2014) was used to analyze the eye movement data. We modeled each participant's eye movement pattern with an HMM, and clustered these HMMs into two general patterns according to their similarities; one of the resulting patterns resembled the central (holistic) pattern, whereas the other resembled the triangular (analytic) pattern (Figure 1). We then calculated the log likelihoods of each participant's eye movement pattern belonging to the two patterns with and without priming. The results showed that compared with the baseline (no priming) condition, local priming significantly increased the likelihood of participants' eye movement pattern being classified as the analytic pattern, whereas global priming did not significantly change the likelihood (Figure 2). This effect can also be seen in fixation maps (Figure 3), although it is relatively harder to interpret the differences. In addition, participants had better recognition performance after local priming than global priming (Figure 4), suggesting an advantage of the analytic strategy in face recognition. Thus, our results suggest that the holistic pattern is linked to engagement of global attention/global information processing, whereas the analytic pattern represents the use of local attention/featural-based processing. Local priming helps participants direct attention to facial features and may consequently enhance recognition performance. Meeting abstract presented at VSS 2015

Mental rotation, as a covert simulation of motor rotation, could benefit from spatial updating of object representations. We are interested in what kind of visual cue could trigger spatial updating. Three experiments were conducted to examine the effect of dynamic and static orientation cues on mental rotation, using a sequential matching task with three-dimensional novel objects presented in different views. Experiment 1 showed that a rotating orientation cue with constant speed reduced viewpoint costs in mental rotation. Experiment 2 extended this effect with a varied-speed rotating orientation cue. However, no such benefit was observed with a static orientation cue in Experiment 3. These findings indicated that a visually continuous orientation cue is sufficient to elicit spatial updating in mental rotation. Furthermore, there may be differences in the underlying mechanisms of spatial updating on the basis of constant-speed rotating cues and varied-speed rotating cues.

Genetic Engineering & Biotechnology NewsVol. 32, No. 7 Assay TutorialLow Variability ADCC BioassayNovel NFAT Reporter Bioassay for Fc Effector FunctionPublished Online:31 Mar 2012https://doi.org/10.1089/gen.32.7.10AboutSectionsView articleView Full TextPDF/EPUB ToolsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail View articleFiguresReferencesRelatedDetailsCited ByAntibody-dependent NK cell degranulation as a marker for assessing antibody-dependent cytotoxicity against pandemic 2009 influenza A(H1N1) infection in human plasma and influenza-vaccinated transchromosomic bovine intravenous immunoglobulin therapyJournal of Virological Methods, Vol. 248 Volume 32Issue 7Apr 2012 Information© 2012 by GEN PublishingTo cite this article:Teresa Surowy, Zhi-Jie (Jey) Cheng, Denise Garvin, Rich Moravec, Aileen Paguio, Neal Cosby, Frank Fan.Low Variability ADCC Bioassay.Genetic Engineering & Biotechnology News.Apr 2012.28-29.http://doi.org/10.1089/gen.32.7.10Published in Volume: 32 Issue 7: March 31, 2012PDF download

Primary peripheral blood mononuclear cells (PBMCs) are routinely used in traditional bioassays to quantify antibody drug potency in antibody-dependent cellular cytotoxicity (ADCC). These bioassays are labor intensive and have high inherent assay variability. Here, we report the development of a bioluminescent cell-based bioassay which measures activation of effector cells via cross-linking of FcαRIIIA with target cell-bound antibodies. For this, Jurkat T-cell stable cell lines that stably express NFAT-luciferase reporter and human FcαRIIIA were generated to replace primary PBMCs as effector cells in ADCC bioassay. Effector cells were also developed in a frozen, thaw-and-use format, to minimize assay variability due to cell culture and handling. The resultant bioassay using this format demonstrated good assay precision and accuracy in bioassay qualification. Bioassay using the engineered effector cells is robust, specific and is able to quantify the potencies of rituximab and trastuzumab, two monoclonal antibody drugs for cancer. When used to measure effects of Fc glycosylation on effector functions of therapeutic antibodies, a linear correlation was observed between relative antibody activity and the extent of Fc glycosylation. Thus, the bioluminescent cell-based reporter bioassay provides a simple and robust approach to measure potency of therapeutic antibodies in ADCC with high precision. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2840. doi:1538-7445.AM2012-2840

The type of experience involved with an object category has been regarded as one important factor in shaping of the human object recognition system. Laboratory training studies have shown that different kinds of learning experience with the same set of novel objects resulted in different perceptual and neural changes. Whether this applies to natural real-world objects remains to be seen. We compared two groups of observers who had different learning experiences with faces, using holistic processing as a dependent measure. We found that, while ordinary observers had extensive individuation experience with faces and displayed typical holistic face processing, art students who had acquired additional experience in drawing faces, and thus in attending to parts of a face, showed less holistic processing than did ordinary observers. These results converge with laboratory training studies on the role of type of experience in the development of different perceptual markers for different object categories. It is thus insufficient to categorize expertise simply in terms of object domains (e.g., expertise with faces). Instead, perceptual expertise should be classified in terms of the underlying process or task demand.

Several clathrin-independent endocytosis mechanisms have been identified that can be distinguished by specific requirements for certain proteins, such as caveolin-1 (Cav1) and the Rho GTPases, RhoA and Cdc42, as well as by specific cargo. Some endocytic pathways may be co-regulated such that disruption of one pathway leads to the up-regulation of another; however, the underlying mechanisms for this are unclear. Cav1 has been reported to function as a guanine nucleotide dissociation inhibitor (GDI), which inhibits Cdc42 activation. We tested the hypothesis that Cav1 can regulate Cdc42-dependent, fluid phase endocytosis. We demonstrate that Cav1 overexpression decreases fluid phase endocytosis, whereas silencing of Cav1 enhances this pathway. Enhancement of Cav1 phosphorylation using a phosphatase inhibitor reduces Cdc42-regulated pinocytosis while stimulating caveolar endocytosis. Fluid phase endocytosis was inhibited by expression of a putative phosphomimetic mutant, Cav1-Y14E, but not by the phospho-deficient mutant, Cav1-Y14F. Overexpression of Cav2, or a Cav1 mutant in which the GDI region was altered to the corresponding sequence in Cav2, did not suppress fluid phase endocytosis. These results suggest that the Cav1 expression level and phosphorylation state regulates fluid phase endocytosis via the interaction between the Cav1 GDI region and Cdc42. These data define a novel molecular mechanism for co-regulation of two distinct clathrin-independent endocytic pathways.

Insulin stimulates glucose transport in fat and skeletal muscle cells primarily by inducing the translocation of GLUT4 (glucose transporter isoform 4) to the PM (plasma membrane) from specialized GSVs (GLUT4 storage vesicles). Glycosphingolipids are components of membrane microdomains and are involved in insulin-regulated glucose transport. Cellular glycosphingolipids decrease during adipocyte differentiation and have been suggested to be involved in adipocyte function. In the present study, we investigated the role of glycosphingolipids in regulating GLUT4 translocation. We decreased glycosphingolipids in 3T3-L1 adipocytes using glycosphingolipid synthesis inhibitors and investigated the effects on GLUT4 translocation using immunocytochemistry, preparation of PM sheets, isolation of GSVs and FRAP (fluorescence recovery after photobleaching) of GLUT4-GFP (green fluorescent protein) in intracellular structures. Glycosphingolipids were located in endosomal vesicles in pre-adipocytes and redistributed to the PM with decreased expression at day 2 after initiation of differentiation. In fully differentiated adipocytes, depletion of glycosphingolipids dramatically accelerated insulin-stimulated GLUT4 translocation. Although insulin-induced phosphorylation of IRS (insulin receptor substrate) and Akt remained intact in glycosphingolipid-depleted cells, both in vitro budding of GLUT4 vesicles and FRAP of GLUT4-GFP on GSVs were stimulated. Glycosphingolipid depletion also enhanced the insulin-induced translocation of VAMP2 (vesicle-associated membrane protein 2), but not the transferrin receptor or cellubrevin, indicating that the effect of glycosphingolipids was specific to VAMP2-positive GSVs. Our results strongly suggest that decreasing glycosphingolipid levels promotes the formation of GSVs and, thus, GLUT4 translocation. These studies provide a mechanistic basis for recent studies showing that inhibition of glycosphingolipid synthesis improves glycaemic control and enhances insulin sensitivity in animal models of Type 2 diabetes.

The G protein coupled receptors (GPCR) represent the target class for nearly half of the current therapeutic drugs and remain to be the focus of drug discovery efforts. The complexity of receptor signaling continues to evolve. It is now known that many GPCRs are coupled to multiple G-proteins, which lead to regulation of respective signaling pathways downstream. Deciphering this receptor coupling will aid our understanding of the GPCR function and ultimately developing drug candidates. Here, we report the development of four homogenous bioluminescent reporter assays using improved destabilized luciferases and various response elements: CRE, NFAT-RE, SRE, and SRF-RE. These assays allowed measurement of major GPCR pathways including cAMP production, intracellular Ca(2+) mobilizations, ERK/MAPK activ-ity, and small G protein RhoA activity, respectively using the same reporter assay format. We showed that we can decipher G protein activation profiles for exogenous m(3) muscarinic receptor and endogenous β(2)-adrenergic receptors in HEK293 cells by using these four reporter assays. Furthermore, we demonstrated that these assays can be readily used for potency rankings of agonists and antagonists, and for high throughput screening.