Standards in Library Technology

published or submitted for publicatio

Papers, Silas and Thomas Orne, 1850-1853

Business papers of Captain Silas M. Orne and Thomas Orne, primarily bills of lading and invoices for wharfage paid for the Schooner Champion out of Boothbay (Me.) and supplies bought, including quantities of salt, as well as fish sold, including cod, polluck, halibut, haddock and hake

Papers, Silas and Thomas Orne, 1854-1856

Business papers of Captain Silas M. Orne and Thomas Orne, primarily bills of lading and invoices for supplies bought, including quantities of salt, and fish sold, including cod, polluck, halibut, haddock and hake, as well as wharfage paid for the Schooners Champion, Ocean, George W. Reed, and Mary A. out of ports in Boothbay (Me.) and Damariscotta (Me.)

Coping Styles of Maltreated Children as Related to Risk and Temperament

A large number of children are classified as maltreated, and these children respond to maltreatment in different ways. Cumulative sociodemographic risk factors and temperament both affect the socioemotional outcomes, including internalizing and externalizing behavior problems. The purpose of this study was to determine whether the association between risk factors and behavioral outcomes in children who have been neglected or abused is influenced by temperamental characteristics. Social workers in Virginia completed questionnaires about five children and adolescents who are part of their current case load. Questionnaires included demographic questions, a Child Behavior Checklist (CBCL), and an Emotionality Activity Sociability (EAS) temperament assessment of the children. This study has practical applications in the area of intervention, as children with high levels of Emotionality show more behavior problems and may need intervention sooner than others

In The Harbor of Your Love

https://digitalcommons.library.umaine.edu/mmb-me/1336/thumbnail.jp

Always, Harvard, Always : We Will Fight For You

https://digitalcommons.library.umaine.edu/mmb-me/1003/thumbnail.jp

Vaccinogenic and Therapeutic Applications of Burkholderia Polysaccharides

Burkholderia pseudomallei the causative agent of melioidosis, is a Gram-negative, facultative-intracellular bacterium that can cause disease in humans and animals. The diagnosis and treatment of melioidosis can be difficult, and at present no licensed vaccines currently exist. Previous studies in our laboratory have revealed that melioidosis subunit vaccine candidates consisting of a B. pseudomallei capsular polysaccharide (CPS)-based glycoconjugate, CPS-CRM197, co-formulated with recombinant B. pseudomallei hemolysin co-regulated protein 1 (Hcp1) or TssM provide high-level protection against acute inhalational challenges of mice with B. pseudomallei. Extending upon these findings in Chapters 2 and 3, we set out to investigate the potential use of Hcp1 and TssM as novel carrier proteins for developing next-generation polysaccharide-based glycoconjugates to immunize against melioidosis. Additionally, we sought to evaluate the potential of FliC and an active-site mutant of the AhpC protein, AhpCC57G, as novel carrier proteins. To facilitate these studies, recombinant Hcp1, TssM, and FliC engineered to possess a single C-terminal cysteine residue (designated Hcp1-Cys, TssM-Cys, and Hcp1-Cys) and the active-site mutant protein AhpCC57G were expressed in Escherichia coli and purified using tandem nickel-cobalt affinity chromatography. Once purified, the cysteine-modified proteins and AhpCC57G were labeled at their C-terminal cysteines using various biotin compounds to confirm the proteins could be modified at the cysteine residues. Following this, Hcp1-Cys and TssM-Cys were modified with heterofunctional linkers to enable site-specific, single-point attachment of CPS or O-polysaccharide (OPS) to the proteins. The glycoconjugates generated in this study were characterized using SDS-PAGE and Western immunoblotting. To assess the immunogenic potential of the glycoconjugates, C57BL/6 mice were immunized with the various glycoconjugates produced in these studies, following which humoral and cellular responses were assessed by ELISA and ELISpot assays, respectively. Results of these studies indicated that the glycoconjugates stimulated the production of high-titer antigen-specific IgG responses. Furthermore, we found that robust interferon (IFN)-γ secreting T-cell responses were raised against the carrier proteins, TssM-Cys and Hcp1-Cys. Finally, we found that the most immunogenic glycoconjugate generated during these studies, OPS-Hcp1, was capable of conferring significant protection in an aerosol challenge with B. pseudomallei K96243. Collectively, our findings suggest that the production of glycoconjugates using site-specific, single-point attachment of Burkholderia-expressed polysaccharides to novel carrier proteins may represent a promising approach for developing a safe and effective melioidosis vaccine comprised exclusively of Burkholderia-specific antigens. In Chapter 4, we sought to develop a process for the isolation, production, and characterization of monoclonal antibodies (mAbs) against CPS for the development of an immunotherapeutic for pre- and post-exposure prophylaxis of melioidosis. B. pseudomallei expresses a variety of structurally conserved protective antigens that are being pursued by our lab as potential vaccine candidates and therapeutic targets. In the present study, we explored the use of 10x Genomics technology to enable the rapid identification and production of CPS-specific mAbs. To facilitate these studies, purified CPS was chemically activated and covalently linked to the carrier protein CRM197 to form the immunogenic glycoconjugate, CPS-CRM197. Mice were then immunized with the glycoconjugate material following which regional lymph nodes were harvested. To isolate B cells from the lymphoid tissues, single-cell suspensions were prepared and depleted of non-target cells using magnetic separation. The enriched B cells were then depleted of naïve IgM+/IgD+ cells resulting in a population of activated B cells that were subjected to 10x single-cell RNA sequencing (scRNAseq). Paired heavy- and light-chain sequences associated with high-frequency clonotypes were synthesized and cloned into expression vectors enabling the production of recombinant mAbs by CHO cells. Using this approach, eight mAbs were purified and characterized by ELISA. The results of these analyses demonstrated that six of the eight recombinant mAbs reacted strongly with purified CPS. The most reactive mAb identified, LN2-3, was further evaluated for potential as an immunotherapeutic. The LN2-3 mAb was successfully subclass-switched to recombinant murine mAbs with different constant regions as well as human chimeric antibodies (ximAbs) for further evaluation and characterization using Western immunoblotting and opsonophagocytosis (OPC) assays. The recombinant murine LN2-3 mAb constructs LN2-3 IgG1 and LN2-3 IgG2c assessed in these studies were capable of conferring protection against an aerosol challenge after passive immunization that was greatly augmented with the administration of co-trimoxazole. Collectively, our findings demonstrate that scRNAseq of activated B cells from mouse lymph nodes represents a promising strategy for the rapid identification and production of antigen-specific mAbs with the potential to serve as an adjunct therapy for the treatment of melioidosis

Une gestion participative de l'eau?: La politique sud-africaine de gestion locale de l'eau

The 1998 South African water reform is a good example of an attempt to democratize water resource management. It created new decentralised water management bodies and openly called for the participation of all individual water users. Yet, if the reform and discourses of the time unequivocally declared the intentions of the South African water law, the conditions surrounding the implementation of the reform left many grey areas in the materialisation of active user participation objectives, almost fifteen years after their adoption. The case of the small-scale irrigation schemes developed in the country's former Bantustans is particularly worrying.La réforme sur l'eau sud-africaine de 1998 est un bon exemple de tentative de démocratisation de la gestion des ressources en eau. Elle crée de nouvelles instances décentralisées de gestion de l'eau et appelle ouvertement à la participation de l'ensemble des usagers individuels de l'eau. Pour autant, si le texte de la réforme et les discours de l'époque affirment sans équivoque les intentions du législateur sud-africain, les conditions de mise en oeuvre de la réforme laissent de nombreuses zones d'ombre quant à la concrétisation à l'échelle locale des objectifs de participation active des usagers près de quinze ans après leur adoption. Le cas des agriculteurs des petits périmètres irrigués créés dans les anciens homelands du pays est particulièrement préoccupant

Vaccinogenic and Therapeutic Applications of Burkholderia Polysaccharides

Burkholderia pseudomallei the causative agent of melioidosis, is a Gram-negative, facultative-intracellular bacterium that can cause disease in humans and animals. The diagnosis and treatment of melioidosis can be difficult, and at present no licensed vaccines currently exist. Previous studies in our laboratory have revealed that melioidosis subunit vaccine candidates consisting of a B. pseudomallei capsular polysaccharide (CPS)-based glycoconjugate, CPS-CRM197, co-formulated with recombinant B. pseudomallei hemolysin co-regulated protein 1 (Hcp1) or TssM provide high-level protection against acute inhalational challenges of mice with B. pseudomallei. Extending upon these findings in Chapters 2 and 3, we set out to investigate the potential use of Hcp1 and TssM as novel carrier proteins for developing next-generation polysaccharide-based glycoconjugates to immunize against melioidosis. Additionally, we sought to evaluate the potential of FliC and an active-site mutant of the AhpC protein, AhpCC57G, as novel carrier proteins. To facilitate these studies, recombinant Hcp1, TssM, and FliC engineered to possess a single C-terminal cysteine residue (designated Hcp1-Cys, TssM-Cys, and Hcp1-Cys) and the active-site mutant protein AhpCC57G were expressed in Escherichia coli and purified using tandem nickel-cobalt affinity chromatography. Once purified, the cysteine-modified proteins and AhpCC57G were labeled at their C-terminal cysteines using various biotin compounds to confirm the proteins could be modified at the cysteine residues. Following this, Hcp1-Cys and TssM-Cys were modified with heterofunctional linkers to enable site-specific, single-point attachment of CPS or O-polysaccharide (OPS) to the proteins. The glycoconjugates generated in this study were characterized using SDS-PAGE and Western immunoblotting. To assess the immunogenic potential of the glycoconjugates, C57BL/6 mice were immunized with the various glycoconjugates produced in these studies, following which humoral and cellular responses were assessed by ELISA and ELISpot assays, respectively. Results of these studies indicated that the glycoconjugates stimulated the production of high-titer antigen-specific IgG responses. Furthermore, we found that robust interferon (IFN)-γ secreting T-cell responses were raised against the carrier proteins, TssM-Cys and Hcp1-Cys. Finally, we found that the most immunogenic glycoconjugate generated during these studies, OPS-Hcp1, was capable of conferring significant protection in an aerosol challenge with B. pseudomallei K96243. Collectively, our findings suggest that the production of glycoconjugates using site-specific, single-point attachment of Burkholderia-expressed polysaccharides to novel carrier proteins may represent a promising approach for developing a safe and effective melioidosis vaccine comprised exclusively of Burkholderia-specific antigens. In Chapter 4, we sought to develop a process for the isolation, production, and characterization of monoclonal antibodies (mAbs) against CPS for the development of an immunotherapeutic for pre- and post-exposure prophylaxis of melioidosis. B. pseudomallei expresses a variety of structurally conserved protective antigens that are being pursued by our lab as potential vaccine candidates and therapeutic targets. In the present study, we explored the use of 10x Genomics technology to enable the rapid identification and production of CPS-specific mAbs. To facilitate these studies, purified CPS was chemically activated and covalently linked to the carrier protein CRM197 to form the immunogenic glycoconjugate, CPS-CRM197. Mice were then immunized with the glycoconjugate material following which regional lymph nodes were harvested. To isolate B cells from the lymphoid tissues, single-cell suspensions were prepared and depleted of non-target cells using magnetic separation. The enriched B cells were then depleted of naïve IgM+/IgD+ cells resulting in a population of activated B cells that were subjected to 10x single-cell RNA sequencing (scRNAseq). Paired heavy- and light-chain sequences associated with high-frequency clonotypes were synthesized and cloned into expression vectors enabling the production of recombinant mAbs by CHO cells. Using this approach, eight mAbs were purified and characterized by ELISA. The results of these analyses demonstrated that six of the eight recombinant mAbs reacted strongly with purified CPS. The most reactive mAb identified, LN2-3, was further evaluated for potential as an immunotherapeutic. The LN2-3 mAb was successfully subclass-switched to recombinant murine mAbs with different constant regions as well as human chimeric antibodies (ximAbs) for further evaluation and characterization using Western immunoblotting and opsonophagocytosis (OPC) assays. The recombinant murine LN2-3 mAb constructs LN2-3 IgG1 and LN2-3 IgG2c assessed in these studies were capable of conferring protection against an aerosol challenge after passive immunization that was greatly augmented with the administration of co-trimoxazole. Collectively, our findings demonstrate that scRNAseq of activated B cells from mouse lymph nodes represents a promising strategy for the rapid identification and production of antigen-specific mAbs with the potential to serve as an adjunct therapy for the treatment of melioidosis

Down At The Summer Resort

https://digitalcommons.library.umaine.edu/mmb-vp/1336/thumbnail.jp