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1.
CBE Life Sci Educ ; 10(4): 342-5, 2011.
Article in English | MEDLINE | ID: mdl-22135368

ABSTRACT

To transform undergraduate biology education, faculty need to provide opportunities for students to engage in the process of science. The rise of research approaches using next-generation (NextGen) sequencing has been impressive, but incorporation of such approaches into the undergraduate curriculum remains a major challenge. In this paper, we report proceedings of a National Science Foundation-funded workshop held July 11-14, 2011, at Juniata College. The purpose of the workshop was to develop a regional research coordination network for undergraduate biology education (RCN/UBE). The network is collaborating with a genome-sequencing core facility located at Pennsylvania State University (University Park) to enable undergraduate students and faculty at small colleges to access state-of-the-art sequencing technology. We aim to create a database of references, protocols, and raw data related to NextGen sequencing, and to find innovative ways to reduce costs related to sequencing and bioinformatics analysis. It was agreed that our regional network for NextGen sequencing could operate more effectively if it were partnered with the Genome Consortium for Active Teaching (GCAT) as a new arm of that consortium, entitled GCAT-SEEK(quence). This step would also permit the approach to be replicated elsewhere.


Subject(s)
Education, Medical, Undergraduate/methods , Genome/genetics , Teaching/methods , Computational Biology/economics , Computational Biology/education , Computational Biology/instrumentation , Congresses as Topic , Databases, Genetic , Educational Technology/economics , Educational Technology/education , Educational Technology/instrumentation , Faculty, Medical/organization & administration , Humans , Oligonucleotide Array Sequence Analysis/instrumentation , Oligonucleotide Array Sequence Analysis/methods , Sequence Analysis, DNA/economics , Sequence Analysis, DNA/instrumentation , Sequence Analysis, DNA/methods , Students, Medical
2.
J Virol ; 80(3): 1340-51, 2006 Feb.
Article in English | MEDLINE | ID: mdl-16415011

ABSTRACT

The flavivirus nonstructural protein NS1 is a highly conserved secreted glycoprotein that does not package with the virion. Immunization with NS1 elicits a protective immune response against yellow fever, dengue, and tick-borne encephalitis flaviviruses through poorly defined mechanisms. In this study, we purified a recombinant, secreted form of West Nile virus (WNV) NS1 glycoprotein from baculovirus-infected insect cells and generated 22 new NS1-specific monoclonal antibodies (MAbs). By performing competitive binding assays and expressing truncated NS1 proteins on the surface of yeast (Saccharomyces cerevisiae) and in bacteria, we mapped 21 of the newly generated MAbs to three NS1 fragments. Prophylaxis of C57BL/6 mice with any of four MAbs (10NS1, 14NS1, 16NS1, and 17NS1) strongly protected against lethal WNV infection (75 to 95% survival, respectively) compared to saline-treated controls (17% survival). In contrast, other anti-NS1 MAbs of the same isotype provided no significant protection. Notably, 14NS1 and 16NS1 also demonstrated marked efficacy as postexposure therapy, even when administered as a single dose 4 days after infection. Virologic analysis showed that 17NS1 protects at an early stage in infection through a C1q-independent and Fc gamma receptor-dependent pathway. Interestingly, 14NS1, which maps to a distinct region on NS1, protected through a C1q- and Fc gamma receptor-independent mechanism. Overall, our data suggest that distinct regions of NS1 can elicit protective humoral immunity against WNV through different mechanisms.


Subject(s)
Antibodies, Monoclonal/pharmacology , Antibodies, Viral/administration & dosage , Receptors, IgG/metabolism , Viral Nonstructural Proteins/immunology , West Nile Fever/immunology , West Nile Fever/prevention & control , West Nile virus/immunology , West Nile virus/pathogenicity , Aedes , Animals , Antigens, Viral/genetics , Base Sequence , Cell Line , Cricetinae , DNA, Viral/genetics , Epitope Mapping , Mice , Mice, Inbred C57BL , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Viral Nonstructural Proteins/genetics , West Nile Fever/therapy , West Nile virus/genetics
3.
J Virol ; 77(24): 12941-9, 2003 Dec.
Article in English | MEDLINE | ID: mdl-14645550

ABSTRACT

West Nile virus (WNV) is a mosquito-borne Flavivirus that causes encephalitis in a subset of susceptible humans. Current treatment for WNV infections is supportive, and no specific therapy or vaccine is available. In this study, we directly tested the prophylactic and therapeutic efficacy of polyclonal antibodies against WNV. Passive administration of human gamma globulin or mouse serum prior to WNV infection protected congenic wild-type, B-cell-deficient ( micro MT), and T- and B-cell-deficient (RAG1) C57BL/6J mice. Notably, no increased mortality due to immune enhancement was observed. Although immune antibody completely prevented morbidity and mortality in wild-type mice, its effect was not durable in immunocompromised mice: many micro MT and RAG1 mice eventually succumbed to infection. Thus, antibody by itself did not completely eliminate viral reservoirs in host tissues, consistent with an intact cellular immune response being required for viral clearance. In therapeutic postexposure studies, human gamma globulin partially protected against WNV-induced mortality. In micro MT mice, therapy had to be initiated within 2 days of infection to gain a survival benefit, whereas in the wild-type mice, therapy even 5 days after infection reduced mortality. This time point is significant because between days 4 and 5, WNV was detected in the brains of infected mice. Thus, passive transfer of immune antibody improves clinical outcome even after WNV has disseminated into the central nervous system.


Subject(s)
Antibodies, Viral/therapeutic use , Immune Sera/administration & dosage , Immunization, Passive , West Nile Fever/drug therapy , West Nile Fever/prevention & control , Animals , Antibodies, Viral/immunology , B-Lymphocytes/immunology , Disease Models, Animal , Humans , Immune Sera/immunology , Immunocompetence , Immunocompromised Host , Infusions, Parenteral , Mice , Mice, Congenic , Mice, Inbred BALB C , T-Lymphocytes/immunology , Treatment Outcome , West Nile Fever/immunology , West Nile Fever/mortality , West Nile virus/immunology
4.
J Physiol ; 542(Pt 1): 237-44, 2002 Jul 01.
Article in English | MEDLINE | ID: mdl-12096065

ABSTRACT

Surfactant-like particles (SLP) are secreted from enterocytes basolaterally into the lamina propria, and reach the apical surface through the intercellular tight junctions. Interactions of SLP with apical and basolateral membranes and with extracellular matrix proteins were measured using a solid-phase binding assay and gel overlays. Small-intestinal SLP bound to basolateral membranes much more than to apical membranes, and more tightly to fibronectin than to laminin (affinity constant K(a) = 1.23 x 10(-2) microg vs. 0.67 x 10(-2) microg; maximal number of binding sites 4.1 microg x ml(-1) vs. 0.32 microg x ml(-1)), but did not bind to collagen types I or IV. Small-intestinal SLP bound fibronectin more than colonic or gastric SLP. Binding to fibronectin was inhibited only partially by RGD peptide and gelatin, but not by heparin. An antibody against alpha(v) integrin also identified the fibronectin-binding component in SLP at approximately 220 kDa, which is the expected size for integrin heterodimers. SLP binding to apical microvillous membranes was weaker and was inhibited by heparin. SLP bound more strongly to heparin itself, and this binding was inhibited by glucuronic acid and chondroitin sulfate. These data are consistent with the hypothesis that the time spent by secreted SLP in the lamina propria is prolonged by strong interactions with proteins in the basolateral membranes, and in the intestinal lumen by weaker interactions with apical membrane components, including heparin. These interactions may allow SLP the time to exert their functions in each tissue compartment.


Subject(s)
Extracellular Matrix Proteins/metabolism , Animals , Blotting, Western , Cell Membrane/drug effects , Cell Membrane/metabolism , Colon/metabolism , Enzyme-Linked Immunosorbent Assay , Epithelial Cells/drug effects , Epithelial Cells/metabolism , Fibronectins/metabolism , Gelatin/metabolism , Gelatin/pharmacology , Heparin/metabolism , Heparin/pharmacology , Integrin alphaV/metabolism , Intestinal Mucosa/cytology , Intestinal Mucosa/drug effects , Intestinal Mucosa/metabolism , Male , Membrane Proteins/metabolism , Microvilli/metabolism , Protein Binding/drug effects , Rats , Triglycerides/metabolism
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