Biomanufacturing and testbed development for the continuous production of monoclonal antibodies

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S110, a novel decitabine dinucleotide, increases fetal hemoglobin levels in baboons (P. anubis)

<p>Abstract</p> <p>Background</p> <p>S110 is a novel dinucleoside analog that could have advantages over existing DNA methyltransferase (DNMT) inhibitors such as decitabine. A potential therapeutic role for S110 is to increase fetal hemoglobin (HbF) levels to treat β-hemoglobinopathies. In these experiments the effect of S110 on HbF levels in baboons and its ability to reduce DNA methylation of the γ-globin gene promoter in vivo were evaluated.</p> <p>Methods</p> <p>The effect of S110 on HbF and γ-globin promoter DNA methylation was examined in cultured human erythroid progenitors and in vivo in the baboon pre-clinical model. S110 pharmacokinetics was also examined in the baboon model.</p> <p>Results</p> <p>S110 increased HbF and reduced DNA methylation of the γ-globin promoter in human erythroid progenitors and in baboons when administered subcutaneously. Pharmacokinetic analysis was consistent with rapid conversion of S110 into the deoxycytosine analog decitabine that binds and depletes DNA.</p> <p>Conclusion</p> <p>S110 is rapidly converted into decitabine, hypomethylates DNA, and induces HbF in cultured human erythroid progenitors and the baboon pre-clinical model.</p

Novel Inducers of Fetal Globin Identified through High Throughput Screening (HTS) Are Active In Vivo in Anemic Baboons and Transgenic Mice

We thank Sarah Haigh, Ada Kane, Nicole Reuter, David Carey, and Marilyn Perry Carey for dedicated and expert technical assistance and Cloret Carl for assistance with preparation of the manuscript.This work was supported by grants from the National Institutes of Health, R01 DK-52962, (SPP, Boston University), R41 HL-105816 (SPP, Phoenicia BioSciences), and R42 HL-110727 (Phoenicia BioSciences), 2 P40 ODO010988-16 (GLW, University of Oklahoma) and UL1-TR000157 (RFW, University of Oklahoma). SMN was supported by P50 HL-118006. The funders had no role in study design, data collection or analysis, decision to publish, or preparation of the manuscript.High-level fetal (γ) globin expression ameliorates clinical severity of the beta (β) hemoglobinopathies, and safe, orally-bioavailable γ-globin inducing agents would benefit many patients. We adapted a LCR-γ-globin promoter-GFP reporter assay to a high-throughput robotic system to evaluate five diverse chemical libraries for this activity. Multiple structurally- and functionally-diverse compounds were identified which activate the γ-globin gene promoter at nanomolar concentrations, including some therapeutics approved for other conditions. Three candidates with established safety profiles were further evaluated in erythroid progenitors, anemic baboons and transgenic mice, with significant induction of γ-globin expression observed in vivo. A lead candidate, Benserazide, emerged which demonstrated > 20-fold induction of γ-globin mRNA expression in anemic baboons and increased F-cell proportions by 3.5-fold in transgenic mice. Benserazide has been used chronically to inhibit amino acid decarboxylase to enhance plasma levels of L-dopa. These studies confirm the utility of high-throughput screening and identify previously unrecognized fetal globin inducing candidates which can be developed expediently for treatment of hemoglobinopathies.Yeshttp://www.plosone.org/static/editorial#pee

Spelling skill and the effectiveness of rhyme and semantic hints to anagram solutions

The present experiment was designed to investigate anagram solution as a function of spelling ability and type of hint. Forty-eight university undergraduates~ identified as good or poor spellers, were presented with anagrams until 15 were missed. Subjects then memorized a list of words for immediate recall in serial order. Words in the memory list were related phonetically and orthographically, semantically~ or were unrelated to the previously missed anagrams. Following the serial recall task, subjects were presented again with those anagrams previously missed. An interaction was predicted to occur between spelling ability and type of hint when solving anagrams. Rh)me hints were expected to facilitate anagram solving in good spellers, but not in poor spellers. The only significant finding was that good spellers solved more anagrams than did poor spellers. Both types of hints (rhyme and semantic) were ineffective and no interaction was found between spelling ability and type of hint. A possibility of serial order difficulty is discussed

p38 MAP kinase activation mediates gamma-globin gene induction in erythroid progenitors

Objective. Our goal was to determine the role of p38 mitogen-activated protein kinase (MAPK) signaling in fetal hemoglobin (HbF) induction. Two histone deacetylase inhibitors (HDAIs), sodium butyrate (NB), and trichostatin (TSA) and hemin were analyzed. In addition, the effect of direct activation of p38 MAPK on gamma-globin gene activity was studied. Method. Primary erythroid progenitors derived from peripheral blood mononuclear cell and K562 erythroleukemia cells were analyzed. Cells were grown in NB, TSA, hemin, or anisomycin either alone or in the presence of the p38 MAPK inhibitor SB203580. The effects of the various treatments on gamma-globin RNA, HbF, and phosphorylated p38 MAPK levels were measured by RNase protection assay, alkaline denaturation, and Western blot analysis, respectively. A K562 stable line overexpressing constitutively active p38 MAPK was established using MAPK kinase kinase 3 (MKK3) and MKK6, the immediate upstream activators of p38. The direct effect of p38 MAPK overexpression on gamma-globin mRNA synthesis was analyzed. Results. NB and TSA activated p38 MAPK and increased gamma-globin mRNA levels in K562 cells and primary erythroid progenitors. Pretreatment with SB203580 blocked p38 MAPK and gamma-globin gene activation. In contrast, no change in p38 activity was observed with hemin inductions. Direct activation of p38 by anisomycin or constitutive overexpression also increased gamma-globin mRNA in the absence of HbF inducers in wild-type K562 cells and in the MKK stable lines. Conclusion. This study supports a novel role for p38 MAPK in gamma-globin regulation in human erythroid progenitors. (C) 2003 International Society for Experimental Hematology.

Production System Diversification and Livelihood in the Drylands of North Central Mexico

Drylands (arid and semi-arid regions) are important regions in the world; they have been disregarded and considered poor undeveloped regions due to their ecological limitations. Farmers in these regions tend towards diversification of production systems in order to achieve livelihood security, and this phenomenon has not been extensively studied. The objective of this study was to create a typology of the production systems present in the Mexican north central drylands, using variables related to production, socioeconomics, and social capital. 1044 interviews were conducted in the semi-arid region of north central Mexico. Analysis of the data allowed for the observation of nine types of production systems distributed in three groups: Subsistence, commercial, and off-farm income systems. The differences observed within these systems are due to generational gaps, gender differences, market orientation, and social capital. It can be concluded that the diversification of the dryland production systems allows for an understanding of why generic public policies have failed to mitigate poverty in these regions. The implications of the study refer to the reconfiguration of Mexican policies for the development of the drylands

Variabilidad cualitativa y cuantitativa de accesiones de amaranto determinada mediante caracterización morfológica

The classification within the genus Amaranthus has been quite difficult, mainly because highly segregating characters within the populations have been considered for this. The aim of this study was to morphologically characterize Amaranthus spp., germplasm in order to know the existent qualitative and quantitative variability in the INIFAP's collection of the genetic resources program (CEVAMEX). During the spring-summer season, 2009, 155 accessions were established in INIFAP's Mexican Valley Experimental Station in Santa Lucía de Prías, Texcoco, State of Mexico. The morphological characterization was made based on a list of qualitative and quantitative descriptors proposed by Grubben and Van Sloten (1981).All the quantitative studied variables showed high variability, considering the highly significant differences between populations and species. The species A. hypochondriacus was the most frequent with 66.7% of the studied germplasm; other species found were A. cruentus, A. caudatus and A. hybridus. A large variation of colors, sizes and shapes in stem, inflorescence and leaves within each species was observed for the qualitative traits. The amaranth germplasm available today shows a large morphological diversity, due to a high degree of cross-breeding. There is quite an enormous phenotypic variation between and within the species of Amaranthus genus, which might be used for breeding proposes and also to understand the diversity of this crop in Mexico.La clasificación dentro del género Amaranthus ha sido difícil, debido a que se han considerado para tal efecto caracteres con alta segregación dentro de las poblaciones. El objetivo del presente estudio fue caracterizar morfológicamente germoplasma de Amaranthus spp., para conocer tanto la variabilidad cualitativa como la cuantitativa existente en la colección del programa de recursos genéticos del INIFAP (CEVAMEX). Se establecieron 155 accesiones durante el ciclo agrícola primavera-verano, 2009 en el Campo Experimental Valle de México del INIFAP, en Santa Lucía de Prías, Texcoco, Estado de México. La caracterización morfológica se realizó con base en una lista de descriptores tanto cualitativos como cuantitativos propuestos por Grubben y Van Sloten (1981). Todas las variables cuantitativas estudiadas presentaron alta variabilidad, pues presentaron diferencias altamente significativas entre poblaciones y especies. La especie A. hypochondriacus fue la de mayor frecuencia con 66.7% del germoplasma estudiado; otras especies encontradas fueron A. cruentus, A. caudatus y A. hybridus. Para caracteres cualitativos existió una enorme variación de colores, tamaños y formas en tallo, inflorescencia y hojas dentro de cada especie bajo estudio. El germoplasma de amaranto disponible en la actualidad muestra una gran diversidad morfológica, producto de un alto grado de cruzamiento. Existe una enorme variación fenotípica entre y dentro de especies del género Amaranthus, que puede ser aprovechada en el mejoramiento genético, así como para conocer la diversidad de este cultivo en México