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Matsushita H.,Sanford Applied Biosciences L.L.C. | Matsushita H.,Hematech Inc. | Sano A.,Chiyoda Corporation | Sano A.,Hematech Inc. | And 11 more authors.
PLoS ONE | Year: 2014

Towards the goal of producing fully human polyclonal antibodies (hpAbs or hIgGs) in transchromosomic (Tc) cattle, we previously reported that Tc cattle carrying a human artificial chromosome (HAC) comprising the entire unrearranged human immunoglobulin (Ig) heavy-chain (hIGH), kappa-chain (hIGK), and lambda-chain (hIGL) germline loci produced physiological levels of hIgGs when both of the bovine immunoglobulin mu heavy-chains, bIGHM and bIGHML1, were homozygously inactivated (bIGHM-/-, bIGHML1-/-; double knockouts or DKO). However, because endogenous bovine immunoglobulin light chain loci are still intact, the light chains are produced both from the hIGK and hIGL genomic loci on the HAC and from the endogenous bovine kappa-chain (bIGK) and lambda-chain (bIGL) genomic loci, resulting in the production of fully hIgGs (both Ig heavy-chains and light-chains are of human origin: hIgG/hIgκ or hIgG/hIgλ) and chimeric hIgGs (Ig heavy-chains are of human origin while the Ig light-chains are of bovine origin: hIgG/bIgκ or hIgG/bIgλ). To improve fully hIgG production in Tc cattle, we here report the deletion of the entire bIGL joining (J) and constant (C) gene cluster (b IGLJ1-IGLC1 to bIGLJ5-IGLC5) by employing Cre/loxP mediated site-specific chromosome recombination and the production of triple knockout (bIGHM-/-, bIGHML1-/- and bIGL-/-; TKO) Tc cattle. We further demonstrate that bIGL cluster deletion greatly improves fully hIgGs production in the sera of TKO Tc cattle, with 51.3% fully hIgGs (hIgG/hIgκ plus hIgG/hIgλ). © 2014 Matsushita et al. Source

Polejaeva I.A.,Utah State University | Broek D.M.,ViaGen Inc. | Walker S.C.,ViaGen Inc. | Zhou W.,ViaGen Inc. | And 3 more authors.
PLoS ONE | Year: 2013

The objective of this study was to determine whether or not reproductive performance in cattle produced by somatic cell nuclear transfer (SCNT) is significantly different from that of their genetic donors. To address this question, we directed two longitudinal studies using different embryo production procedures: (1) superovulation followed by artificial insemination (AI) and embryo collection and (2) ultrasound-guided ovum pick-up followed by in vitro fertilization (OPU-IVF). Collectively, these two studies represent the largest data set available for any species on the reproductive performance of female clones and their genetic donors as measured by their embryo production outcomes in commercial embryo production program. The large-scale study described herein was conducted over a six-year period of time and provides a unique comparison of 96 clones to the 40 corresponding genetic donors. To our knowledge, this is the first longitudinal study on the reproductive performance of cattle clones using OPU-IVF. With nearly 2,000 reproductive procedures performed and more than 9,200 transferable embryos produced, our observations show that the reproductive performance of cattle produced by SCNT is not different compared to their genetic donors for the production of transferable embryos after either AI followed by embryo collection ( P = 0.77) or OPU-IVF (P = 0.97). These data are in agreement with previous reports showing that the reproductive capabilities of cloned cattle are equal to that of conventionally produced cattle. In conclusion, results of this longitudinal study once again demonstrate that cloning technology, in combination with superovulation, AI and embryo collection or OPU-IVF, provides a valuable tool for faster dissemination of superior maternal genetics. © 2013 Polejaeva et al. Source

Sano A.,Chiyoda Corporation | Sano A.,Hematech Inc. | Matsushita H.,Sanford Applied Biosciences L.L.C. | Matsushita H.,Hematech Inc. | And 12 more authors.
PLoS ONE | Year: 2013

Therapeutic human polyclonal antibodies (hpAbs) derived from pooled plasma from human donors are Food and Drug Administration approved biologics used in the treatment of a variety of human diseases. Powered by the natural diversity of immune response, hpAbs are effective in treating diseases caused by complex or quickly-evolving antigens such as viruses. We previously showed that transchromosomic (Tc) cattle carrying a human artificial chromosome (HAC) comprising the entire unrearranged human immunoglobulin heavy-chain (hIGH) and kappa-chain (hIGK) germline loci (named as κHAC) are capable of producing functional hpAbs when both of the bovine immunoglobulin mu heavy-chains, bIGHM and bIGHML1, are homozygously inactivated (double knockouts or DKO). However, B lymphocyte development in these Tc cattle is compromised, and the overall production of hpAbs is low. Here, we report the construction of an improved HAC, designated as cKSL-HACΔ, by incorporating all of the human immunoglobulin germline loci into the HAC. Furthermore, for avoiding the possible human-bovine interspecies incompatibility between the human immunoglobulin mu chain protein (hIgM) and bovine transmembrane α and β immunoglobulins (bIgα and bIgβ) in the pre-B cell receptor (pre-BCR) complex, we partially replaced (bovinized) the hIgM constant domain with the counterpart of bovine IgM (bIgM) that is involved in the interaction between bIgM and bIgα/Igβ; human IgM bovinization would also improve the functionality of hIgM in supporting B cell activation and proliferation. We also report the successful production of DKO Tc cattle carrying the cKSL-HACΔ (cKSL-HACΔ/DKO), the dramatic improvement of B cell development in these cattle and the high level production of hpAbs (as measured for the human IgG isotype) in the plasma. We further demonstrate that, upon immunization by tumor immunogens, high titer tumor immunogen-specific human IgG (hIgG) can be produced from such Tc cattle. © 2013 Sano et al. Source

News Article
Site: http://www.scientificcomputing.com/rss-feeds/all/rss.xml/all

Austin, TX — Silicon Mechanics, a provider of servers, storage and high-performance computing solutions, announced the opening of its 5th Annual Research Cluster Grant (RCG) program at Supercomputing 2015. Two institutions will be selected, and both will be awarded a complete high-performance computing (HPC) cluster. The competition is open to all United States and Canadian qualified post-secondary institutions, university-affiliated research institutions, non-profit research institutions, and researchers at federal labs with university affiliations. "We designed the Research Cluster Grant program to provide computational and storage technology resources to researchers who may not have been able to keep pace with technology acquisitions through traditional grant-funding programs like those at the National Science Foundation or the National Institute of Health,” said Art Mann, Silicon Mechanics' Sr. Director, Life Sciences Practice. "With the ever-growing demand for more powerful IT infrastructure to support research, the RCG represents a tremendous opportunity to work with our technology partners and support these research efforts. I’m excited and truly honored to see the RCG program achieve its fifth year.” Silicon Mechanics created the RCG in 2012 as a way of giving back to the educational community, as obtaining needed research funding for technology advancements continues to be challenging and can limit future impact at some educational institutions. In particular, the program is helping to jumpstart research efforts where access to high-performance computing is limited, outdated or was not previously available. The RCG program also provides institutions with an opportunity to showcase how collaboration across departments and researchers by providing cluster technology can positively impact research efforts through the use of cluster technology. Previous RCG awardees include The City College of New York (CCNY) and Dordt College in 2015, Wayne State University in 2014, Tufts University in 2013 and Saint Louis University in 2012. Silicon Mechanics' partners currently committed to supporting this year’s grant include: Intel, NVIDIA, Mellanox, Supermicro, Bright Computing, HGST, Avago, Kingston, Micron and Seagate. At CCNY, the HPC cluster is being used for cutting-edge research in biochemistry, chemistry, biology, physics, earth and atmospheric sciences, computer science, engineering, medicine, mathematics, social science, humanities and writing pedagogy. "For many of our research programs, this computer cluster was the missing piece that lowered the barriers that kept our work from moving forward smoothly," said David Jeruzalmi, professor of chemistry and biochemistry in CCNY's Division of Science, who wrote the grant proposal last year. "This award has touched the research of many colleagues by bringing together researchers from across CCNY, many of whom never knew that their work could be positively impacted by colleagues down the hall or in the next building over." At Dordt College and at its research partner, Hope College, the HPC cluster supports eight STEM-based research groups and nine distinct faculty members focused on a wide variety of research activities. Those activities include bacterial statistical genetics, processing and analysis of RNA sequencing, phylogenetic trees, computational chemistry, engineering integrity, analyzing genomic sequencing data, population genetic data and more. "Dordt has traditionally been a liberal arts school," said Dr. Nathan Tintle, Dordt College's Director for Research and Scholarship. "In recent years, however, we have ramped up our research department in partnership with Hope College and, in doing so, created a demand for an HPC system. Unfortunately, we didn’t have the budget to purchase a cluster that would suit our computational needs. Fortunately, Silicon Mechanics offered the annual RCG, a program that we are proud to be involved with. We feel fortunate to have been awarded this grant." Submissions for the 2016 RCG will be accepted December 15, 2015, through March 1, 2016. The grant recipients will be announced April 2016. Submissions will be reviewed for merit and for the potential impact the research may have on the institution's mission. Silicon Mechanics strongly encourages collaboration, within and across departments of a single institution, or across multiple institutions. Details on RCG rules, application requirements, and cluster technical specifications are available at www.researchclustergrant.com. About The City College of New York Since 1847, The City College of New York has provided low-cost, high-quality education for New Yorkers in a wide variety of disciplines. More than 16,000 students pursue undergraduate and graduate degrees in: the College of Liberal Arts and Sciences; the Bernard and Anne Spitzer School of Architecture; the School of Education; the Grove School of Engineering; the Sophie Davis School of Biomedical Education, and the Colin Powell School for Civic and Global Leadership. Dordt College is a private institution of higher education, committed to the Reformed Christian perspective. With 1,400 students, the college’s STEM programs are leading enrollment growth. Located in Sioux Center, Iowa, Dordt College provides a holistic residential learning experience for students, in which they can develop Christian insight in all areas of life.

Zhou W.,ViaGen Inc. | Gosch G.,Sioux Center | Guerra T.,ViaGen Inc. | Broek D.,Sioux Center | And 4 more authors.
Theriogenology | Year: 2014

Somatic cell nuclear transfer (SCNT), or cloning, is one of the assisted reproductive technologies currently used in agriculture. Commercial applications of SCNT are presently limited to the production of animals of high genetic merit or the production of the most elite show cattle owing to its relatively low efficiency. In current practice, 20% to 40% of SCNT pregnancies do not result in viable offspring. In an effort to better understand some of the anomalies associated with SCNT pregnancies, we investigated amino acid compositions of first trimester amniotic fluid. In this retrospective study, amniotic fluids were collected from SCNT and control IVF pregnancies at Day 75 of gestation and grouped according to the pregnancy results: control IVF (IVF), viable SCNT pregnancies that resulted in live healthy calves (SCNT-HL), nonviable SCNT pregnancies that were aborted before Day 150 (SCNT-ED), and nonviable SCNT pregnancies that were aborted after Day 150 or produced deceased calves (SCNT-LD). High-performance liquid chromatography (HPLC) was used to analyze the concentrations of 22 amino acids (AAs) in the amniotic fluid samples. There were no differences in average AA concentrations between IVF and SCNT-HL groups, whereas SCNT-LD and SCNT-ED had higher levels of total AA concentrations. Concentrations of asparagine, citruline, arginine, and valine were significantly higher in the SCNT-LD group. Both SCNT-LD and SCNT-ED groups had relatively large intragroup variances in AA concentrations. Urea concentration was also measured in the SCNT amniotic fluid samples. No correlations between urea concentrations and arginine concentrations or pregnancy outcomes were found. The findings in this study not only deepen the understanding onSCNT pregnancy anomalies, but also provide a potentially useful screening tool for assessing viable and nonviable SCNT pregnancies. © 2014 Elsevier Inc. Source

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