385: Monocyte Blood Count and Acute GVHD: Flow Cytometry Analysis Recommended in Prospective Studies

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A Leu to Ile but not Leu to Val change at HIV-1 reverse transcriptase codon 74 in the background of K65R mutation leads to an increased processivity of K65R+L74I enzyme and a replication competent virus

<p>Abstract</p> <p>Background</p> <p>The major hurdle in the treatment of Human Immunodeficiency virus type 1 (HIV-1) includes the development of drug resistance-associated mutations in the target regions of the virus. Since reverse transcriptase (RT) is essential for HIV-1 replication, several nucleoside analogues have been developed to target RT of the virus. Clinical studies have shown that mutations at RT codon 65 and 74 which are located in β3-β4 linkage group of finger sub-domain of RT are selected during treatment with several RT inhibitors, including didanosine, deoxycytidine, abacavir and tenofovir. Interestingly, the co-selection of K65R and L74V is rare in clinical settings. We have previously shown that K65R and L74V are incompatible and a R→K reversion occurs at codon 65 during replication of the virus. Analysis of the HIV resistance database has revealed that similar to K65R+L74V, the double mutant K65R+L74I is also rare. We sought to compare the impact of L→V <it>versus </it>L→I change at codon 74 in the background of K65R mutation, on the replication of doubly mutant viruses.</p> <p>Methods</p> <p>Proviral clones containing K65R, L74V, L74I, K65R+L74V and K65R+L74I RT mutations were created in pNL4-3 backbone and viruses were produced in 293T cells. Replication efficiencies of all the viruses were compared in peripheral blood mononuclear (PBM) cells in the absence of selection pressure. Replication capacity (RC) of mutant viruses in relation to wild type was calculated on the basis of antigen p24 production and RT activity, and paired analysis by student t-test was performed among RCs of doubly mutant viruses. Reversion at RT codons 65 and 74 was monitored during replication in PBM cells. In vitro processivity of mutant RTs was measured to analyze the impact of amino acid changes at RT codon 74.</p> <p>Results</p> <p>Replication kinetics plot showed that all of the mutant viruses were attenuated as compared to wild type (WT) virus. Although attenuated in comparison to WT virus and single point mutants K65R, L74V and L74I; the double mutant K65R+L74I replicated efficiently in comparison to K65R+L74V mutant. The increased replication capacity of K65R+L74I viruses in comparison to K65R+L74V viruses was significant at multiplicity of infection 0.01 (p = 0.0004). Direct sequencing and sequencing after population cloning showed a more pronounced reversion at codon 65 in viruses containing K65R+L74V mutations in comparison to viruses with K65R+L74I mutations. In vitro processivity assays showed increased processivity of RT containing K65R+L74I in comparison to K65R+L74V RT.</p> <p>Conclusions</p> <p>The improved replication kinetics of K65R+L74I virus in comparison to K65R+L74V viruses was due to an increase in the processivity of RT containing K65R+L74I mutations. These observations support the rationale behind structural functional analysis to understand the interactions among unique RT mutations that may emerge during the treatment with specific drug regimens.</p

An integrative process for Advanced Energy Retrofit Projects

Advanced Energy Retrofit (AER) projects can significantly reduce energy consumption of existing buildings, and studies have indicated that they can be more economic than constructing new buildings. Retrofit projects vary in complexity based on the building characteristics, building area, client requirements, project budget, payback period and energy savings targets. The lack of a transparent and streamlined integrative process makes it difficult for the participants to implement AER projects. To implement comprehensive AER projects (also known as Deep Retrofits) in an efficient manner, it is important to understand the process along with the information exchanged during the planning, design, construction and operation phases. This paper describes the development of an AER integrative delivery process model with a focus on the planning and design phases. The integrative process has been developed through a detailed literature review, and studying the project documents and procedures used to deliver an ongoing comprehensive AER project. It was validated by review in multiple industry workshops and through interview with industry and academic experts, including all the primary team members of a comprehensive AER case study. The process is undergoing additional validation through AER integrative workshops and a detailed case study analysis. The final integrative process includes activities focused upon specific building systems along with integrated processes and information exchanges which support the integrated workflow among team members. The identified important information exchanges outline a series of exchanges that support application interoperability for AER projects. Read More: http://ascelibrary.org/doi/abs/10.1061/9780784413616.033link_to_OA_fulltex

Osteomyelitis of the Mandible Secondary to Impacted Third Molar

Odontogenic infections resulting in osteomyelitis of the mandible are common. However, osteomyelitis of the coronoid process as a direct consequence of third molar infection is rare. The following report describes such a case in an 18-year-old female as a severe complication following third molar removal. Despite aggressive antimicrobial therapy and indicated treatment approach following microbiologic study, a severe osteomyelitis was present, clinically and radiographically. Finally, hemimandibulectomy, including exarticulation, was indicated to manage this severe complication