Use of Phage Display to Isolate Specifi c Human Monoclonal Antibody Fragments Against a Potential Target for Multiple Myeloma

Introduction: Multiple myeloma (MM), a malignancy of plasma cells, accounts for 10% of all haematological malignancies and is currently incurable. Although it can be treated, the disease tends to relapse after several years and becomes increasingly resistant to conventional therapy. Investigations into using humoral therapy for MM are now underway with a view that novel therapeutic agents may provide a more targeted therapy for MM. Materials and Methods: Here, phage display, a faster and more efficient method compared to classical hybridoma fusion technology, was used as a proof-of-concept to isolate several single-chain Fragment variables (scFv) against Ku86. Results: Anti-Ku86 polyclonal scFvs biopanning was successful where third round scFvs (A450~1.1) showed a 1/3 increase in binding as compared to the first round scFvs (A450~0.4) with 100ug/mL of antigen (purified human Ku86). Subsequent selection and verifi cation of monoclonal antibodies using third round biopanning revealed 4 good affi nity binding clones ranging from A450~0.1 to A450~0.15 on 12.5ug/mL of antigen as compared to low binders (A450~0.07) and these antibodies bind to Ku86 in a specifi c and dose-dependent manner. Comparative studies were also performed with commercially available murine antibodies and results suggest that 2 of the clones may bind close to the following epitopes aa506-541 and aa1- 374. Conclusions: These studies using phage display provide an alternative and viable method to screen for antibodies quickly and results show that good affinity antibodies against Ku86 have been successfully isolated and they can be used for further studies on MM and form the basis for further development as anti-cancer therapeutic agents

Use of ultraviolet-light irradiated multiple myeloma cells as immunogens to generate tumor-specific cytolytic T lymphocytes

10.1186/1476-8518-6-2Journal of Immune Based Therapies and Vaccines6

Use of Ultraviolet Light Irradiated Multiple Myeloma Cells as Immunogens to generate Tumor Specific Cytolytic T Lymphocytes

Background: As the eradication of tumor cells in vivo is most efficiently performed by cytolytic Tlymphocytes (CTL), various methods for priming tumor-reactive lymphocytes have been developed. In this study, a method of priming CTLs with ultraviolet (UV)-irradiated tumor cells, which results in termination of tumor cell proliferation, apoptosis, as well as upregulation of heat shock proteins (HSP) expression is described. Methods: Peripheral blood mononuclear cells (PBMC) were primed weekly with UV-irradiated or mitomycin-treated RPMI 8226 multiple myeloma cells. Following three rounds of stimulation over 21 days, the lymphocytes from the mixed culture conditions were analyzed for anti-MM cell reactivity. Results: By day 10 of cultures, PBMCs primed using UV-irradiated tumor cells demonstrated a higher percentage of activated CD8+/CD4- T lymphocytes than non-primed PBMCs or PBMCs primed using mitomycin-treated MM cells. Cytotoxicity assays revealed that primed PBMCs were markedly more effective (p \u3c 0.01) than non-primed PBMCs in killing RPMI 8226 MM cells. Surface expression of glucose regulated protein 94 (Grp94/Gp96) and Grp78 were both found to be induced in UV-treated MM cells. Conclusion: Since, HSP-associated peptides are known to mediate tumor rejection; these data suggest that immune-mediated eradication of MM cells could be elicited via a UV-induced HSP process. The finding that the addition of 17-allylamide-17-demethoxygeldanamycin (17AAG, an inhibitor of HSP 90-peptide interactions) resulted in decreased CTL-induced cytotoxicity supported this hypothesis. Our study, therefore, provides the framework for the development of anti-tumor CTL cellular vaccines for treating MM using UV-irradiated tumor cells as immunogens

Footnotes to Disciples History

From the mid 1950s through the 1990s, DCHS printed short publications to remind its readers about the importance of the past. The series included reprints as well as new historical studies. Footnotes to Disciples History collects this wide-ranging series into one volume. ⁃ Rice Haggard\u27s An Address to the Different Religious Societies, on the sacred import of the Christian Name (1804) ⁃ John M. Imbler\u27s Beyond Buffalo: Alexander Campbell on Education for Ministry (1992) ⁃ Report of the Proceedings of a General Meeting of Messengers from Thirteen Congregations (J.T. McVay and Alexander Campbell, 1834) ⁃ William A. Gerrards\u27s Christian Unity, Our Heritage (1986) ⁃ Alexander Campbell\u27s Lunenburg Letter (1837) ⁃ Eva Jean Wrather\u27s Alexander Campbell and His Relevance for Today (1959) ⁃ Perry Gresham\u27s The Broncho That Would Not Be Broken (1986)https://digitalcommons.acu.edu/acu_library_books/1011/thumbnail.jp

During the summer 2009 outbreak of "swine flu" in Scotland what respiratory pathogens were diagnosed as H1N1/2009?

<p>Abstract</p> <p>Background</p> <p>During the April-July 2009 outbreak of H1N1/2009 in scotland the West of Scotland Specialist Virology Centre (WoSSVC) in Glasgow tested > 16 000 clinical samples for H1N1/2009. Most were from patients clinically diagnosed with H1N1/2009. Out of these, 9% were positive. This study sought to determine what respiratory pathogens were misdiagnosed as cases of H1N1/2009 during this time.</p> <p>Methods</p> <p>We examined the results from 3247 samples which were sent to the laboratory during April-July 2009. All were from patients clinically diagnosed as having H1N1/2009 (based on accepted criteria) and all were given a full respiratory screen using real time reverse transcriptase polymerase chain reaction (rtRT-PCR) assays.</p> <p>Results</p> <p>In total, respiratory pathogens were detected in 27.9% (95% confidence interval, 26.3-29.5%) of the samples submitted. Numerous pathogens were detected, the most common of which were rhinovirus (8.9% (95% confidence interval, 7.9-9.9%)), parainfluenza 1 (1.9% (95% confidence interval, 1.4-2.4%)) and 3 (4.1% (95% confidence interval, 3.3-4.9%)), and adenovirus ((3.5% (95% confidence interval, 2.9-4.2%)).</p> <p>Conclusions</p> <p>This study highlights the problems of using a clinical algorithm to detect H1N1/2009. Clinicians frequently misdiagnosed common respiratory pathogens as H1N1/2009 during the spring/summer outbreak in Scotland. Many undesirable consequences would have resulted, relating to treatment, infection control, and public health surveillance.</p

Potential for La Crosse virus segment reassortment in nature

The evolutionary success of La Crosse virus (LACV, family Bunyaviridae) is due to its ability to adapt to changing conditions through intramolecular genetic changes and segment reassortment. Vertical transmission of LACV in mosquitoes increases the potential for segment reassortment. Studies were conducted to determine if segment reassortment was occurring in naturally infected Aedes triseriatus from Wisconsin and Minnesota in 2000, 2004, 2006 and 2007. Mosquito eggs were collected from various sites in Wisconsin and Minnesota. They were reared in the laboratory and adults were tested for LACV antigen by immunofluorescence assay. RNA was isolated from the abdomen of infected mosquitoes and portions of the small (S), medium (M) and large (L) viral genome segments were amplified by RT-PCR and sequenced. Overall, the viral sequences from 40 infected mosquitoes and 5 virus isolates were analyzed. Phylogenetic and linkage disequilibrium analyses revealed that approximately 25% of infected mosquitoes and viruses contained reassorted genome segments, suggesting that LACV segment reassortment is frequent in nature

Increasing frailty is associated with higher prevalence and reduced recognition of delirium in older hospitalised inpatients: results of a multi-centre study

Purpose: Delirium is a neuropsychiatric disorder delineated by an acute change in cognition, attention, and consciousness. It is common, particularly in older adults, but poorly recognised. Frailty is the accumulation of deficits conferring an increased risk of adverse outcomes. We set out to determine how severity of frailty, as measured using the CFS, affected delirium rates, and recognition in hospitalised older people in the United Kingdom. Methods: Adults over 65 years were included in an observational multi-centre audit across UK hospitals, two prospective rounds, and one retrospective note review. Clinical Frailty Scale (CFS), delirium status, and 30-day outcomes were recorded. Results: The overall prevalence of delirium was 16.3% (483). Patients with delirium were more frail than patients without delirium (median CFS 6 vs 4). The risk of delirium was greater with increasing frailty [OR 2.9 (1.8–4.6) in CFS 4 vs 1–3; OR 12.4 (6.2–24.5) in CFS 8 vs 1–3]. Higher CFS was associated with reduced recognition of delirium (OR of 0.7 (0.3–1.9) in CFS 4 compared to 0.2 (0.1–0.7) in CFS 8). These risks were both independent of age and dementia. Conclusion: We have demonstrated an incremental increase in risk of delirium with increasing frailty. This has important clinical implications, suggesting that frailty may provide a more nuanced measure of vulnerability to delirium and poor outcomes. However, the most frail patients are least likely to have their delirium diagnosed and there is a significant lack of research into the underlying pathophysiology of both of these common geriatric syndromes

Interannual Herbaceous Biomass Response to Increasing Honey Mesquite Cover on Two Soils

This study quantified herbaceous biomass responses to increases in honey mesquite (Prosopis glandulosa Torr.) cover on two soils from 1995 to 2001 in north central Texas. Vegetation was sampled randomly with levels of mesquite ranging from 0% to 100%. With no mesquite covering the silt loam soils of bottomland sites, peak herbaceous biomass averaged (6SE) X 300 +/- 210 kg ha-1 vs. –560 +/- 190 kg ha-1 on clay loam soils of upland sites (P = 0.001). A linear decline of 14 +/- 2.5 kg ha-1 in herbaceous biomass occurred for each percent increase in mesquite cover (P = 0.001). The slope of this decline was similar between soils (P=0.135). Herbaceous biomass with increasing mesquite cover varied between years (P=0.001) as did the slope of decline (P=0.001). Warm-season herbaceous biomass decreased linearly with increasing mesquite cover averaging a 73 +/- 15% reduction at 100% mesquite cover (P = 0.001) compared to 0% mesquite cover. Cool-season herbaceous biomass was similar between soils with no mesquite, 1 070 +/- 144 kg ha-1 for silt loam vs. 930 +/- 140 kg ha-1 for clay loam soils, but averaged 340 +/- 174 kg ha-1 more on silt loam than on clay loam soils at 100% mesquite cover (P = 0.004). Multiple regression analysis indicated that each centimeter of precipitation received from the previous October through the current September produced herbaceous biomass of 51 kg ha-1 on silt loam and 41 kg ha-1 on clay loam soils. Herbaceous biomass decreased proportionally with increasing mesquite cover up to 29 kg ha-1 at 100% mesquite cover for each centimeter of precipitation received from January through September. Increasing mesquite cover reduces livestock forage productivity and intensifies drought effects by increasing annual herbaceous biomass variability. From a forage production perspective there is little advantage to having mesquite present. The Rangeland Ecology & Management archives are made available by the Society for Range Management and the University of Arizona Libraries. Contact [email protected] for further information.Migrated from OJS platform August 202