Experimental Transmission of Kaposi's Sarcoma–Associated Herpesvirus (Kshv/Hhv-8) to Scid-Hu Thy/Liv Mice

Kaposi's sarcoma–associated herpesvirus (KSHV/HHV-8) is a novel human lymphotropic herpesvirus linked to several human neoplasms. To date, no animal model for infection by this virus has been described. We have examined the susceptibility of C.B-17 scid/scid mice implanted with human fetal thymus and liver grafts (SCID-hu Thy/Liv mice) to KSHV infection. KSHV virions were inoculated directly into the implants, and viral DNA and mRNA production was assayed using real-time quantitative polymerase chain reaction. This revealed a biphasic infection, with an early phase of lytic replication accompanied and followed by sustained latency. Ultraviolet irradiation of the inoculum abolished all DNA- and mRNA-derived signals, and infection was inhibited by ganciclovir. Viral gene expression was most abundant in CD19+ B lymphocytes, suggesting that this model faithfully mimics the natural tropism of this virus. Short-term coinfection with HIV-1 did not alter the course of KSHV replication, nor did KSHV alter levels of HIV-1 p24 during the acute phase of the infection. Although no disease was evident in infected animals, SCID-hu Thy/Liv mice should allow the detailed study of KSHV tropism, latency, and drug susceptibility

Virus expression detection reveals RNA-sequencing contamination in TCGA

Background: Contamination of reagents and cross contamination across samples is a long-recognized issue in molecular biology laboratories. While often innocuous, contamination can lead to inaccurate results. Cantalupo et al., for example, found HeLa-derived human papillomavirus 18 (H-HPV18) in several of The Cancer Genome Atlas (TCGA) RNA-sequencing samples. This work motivated us to assess a greater number of samples and determine the origin of possible contaminations using viral sequences. To detect viruses with high specificity, we developed the publicly available workflow, VirDetect, that detects virus and laboratory vector sequences in RNA-seq samples. We applied VirDetect to 9143 RNA-seq samples sequenced at one TCGA sequencing center (28/33 cancer types) over 5 years. Results: We confirmed that H-HPV18 was present in many samples and determined that viral transcripts from H-HPV18 significantly co-occurred with those from xenotropic mouse leukemia virus-related virus (XMRV). Using laboratory metadata and viral transcription, we determined that the likely contaminant was a pool of cell lines known as the "common reference", which was sequenced alongside TCGA RNA-seq samples as a control to monitor quality across technology transitions (i.e. microarray to GAII to HiSeq), and to link RNA-seq to previous generation microarrays that standardly used the "common reference". One of the cell lines in the pool was a laboratory isolate of MCF-7, which we discovered was infected with XMRV; another constituent of the pool was likely HeLa cells. Conclusions: Altogether, this indicates a multi-step contamination process. First, MCF-7 was infected with an XMRV. Second, this infected cell line was added to a pool of cell lines, which contained HeLa. Finally, RNA from this pool of cell lines contaminated several TCGA tumor samples most-likely during library construction. Thus, these human tumors with H-HPV or XMRV reads were likely not infected with H-HPV 18 or XMRV

Long-Range N14 Coupling in Ethyl Ammonium Ions

There has been considerable interest recently in the magnitudes and relative signs of CH3-X and CH2-X coupling constants in compounds of the type (CH3CH2)nX [2-6]. We wish to report the analysis of the spectra of N(CH2CH3)4+ and N(C2H5)3(CH2SCH3)+ and the confirmation of the assignments of these spectra by H1-{N14} [1] double resonance and by the temperature dependence of the proton spectra

Influence of antigen distribution on the mediation of immunological glomerular injury

Influence of antigen distribution on the mediation of immunological glomerular injury. To determine if the site of immune reaction could influence the mediation and morphological expression of glomerular injury in experimental anti-glomerular basement membrane (anti-GBM) nephritis and membranous nephropathy, we studied the events that followed the in situ reaction of rat antibody with antigen planted in either the GBM (especially the lamina rara interna) or in the subepithelial space (SE). Non-nephritogenic amounts of noncomplement-fixing sheep anti-GBM or anti-tubular brushborder antibody were injected into separate groups of rats to plant sheep IgG in the GBM and SE, respectively. Kidneys containing sheep IgG were then transplanted into naive recipients that were passively immunized with rat anti-sheep IgG. There was marked proteinuria after 2 days (antigen in GBM: 226 ± 50.7; antigen in SE: 69 ± 50.7 mg/24 hr) that was abrogated by prior depletion of complement in both groups (antigen in GBM: 10.2 ± 1.7; antigen in SE: 14.3 ± 8.7 mg/24 hr). When antigen was planted in SE, inflammatory-cell depletion with either anti-neutrophil (PMN) serum or lethal irradiation had no effect on proteinuria. In contrast, anti-PMN abolished proteinuria (12.0 ± 5.6 mg/24 hr) and irradiation reduced it by 60% when antigen was in GBM. Glomeruli of kidneys with antigen in GBM were significantly larger and more hypercellular than those with antigen in SE after transplantation into immunized recipients. Endothelial cell injury and adherence of inflammatory cells to denuded GBM were prominent in the former (antigen in GBM), while glomeruli with antigen in SE showed only subepithelial deposits, adjacent slit-diaphragm displacement, and epithelial cell foot-process effacement. Thus, the reaction of antigen and antibody in glomeruli produced complement-mediated injury which was cell-independent when complex formation occurred on the outer aspect of the GBM but was cell-dependent when the same reagents reacted more proximally to the circulation. We therefore conclude that antigen distribution can critically influence the mediation and morphologic expression of immune glomerular injury and may, in part, account for variations in the clinical and histological manifestations of antibody-induced glomerular disease in humans.Influence de la distribution antigénique sur la médiation des lésions glomérulaires immunologiques. Afin de déterminer si le site de la réaction immune pourrait influencer la médiation et l'expression morphologique des lésions glomérulaires lors d'une néphrite expérimentale anti-membrane basale glomérulaire (anti-GBM) et d'une néphropathie extra-membraneuse, nous avons étudié les événements qui suivaient la réaction in situ d'anticorps de rat avec un antigène fixé soit dans la GBM (surtout dans la lamina rara interna), soit dans l'espace sous-épithélial (SE). Des quantités non nephritogènes d'anticorps anti-GBM, ou anti-bordure en brosse tubulaire de mouton ne fixant pas le complément ont été injectées à différents groupes de rats pour fixer de l'IgG de mouton dans la GBM et le SE, respectivement. Les reins contenant l'IgG de mouton étaient alors transplantés à des receveurs vierges passivement immunisés avec de l'IgG de rat antimouton. Il existait une protéinurie marquée après deux jours (antigène dans la GBM: 226 ± 50,7; antigène dans SE: 69 ± 50,7 mg/24 hrs) qui à été abrogé par une déplétion du complement dans les deux groupes (antigène dans la GBM: 10,2 ± 1,7; antigène dans SE: 14,3 ± 8,7 mg/24 hr). Lorsque l'antigène était fixé dans SE, une déplétion en cellules inflammatoires par du sérum anti-neutrophile (PMN) ou une irradiation léthale n'avaient pas d'effet sur la protéinurie. A l'opposé, anti-PMN supprimait la protéinurie (12,0 ± 5,6 mg/24 hr) et l'irradiation la réduisait de 60% lorsque l'antigène était dans la GBM. Les glomérules de reins ayant l'antigène dans la GBM étaient significativement plus gros et plus hyper-cellulaires que ceux ayant l'antigène dans SE après transplantation chez des receveurs immunisés. Les lésions cellulaires endothéliales et l'adhérence des cellules inflammatoires à des GBM nues étaient prédominantes chez les premiers (antigène dans la GBM) alors que les glomérules ayant l'antigène dans SE présentaient uniquement des dépôts sous-épithéliaux, un déplacement du slit-diaphragme adjacent et un effacement des pédicelles des cellules épithéliales. Ainsi, la réaction d'un antigène et d'un anticorps dans des glomérules a produit des lésions à médiation complémentaire indépentantes des cellules lorsque la formation de complexes survenait dans la partie extérieure de la GBM, mais dépendantes des cellules lorsque les mêmes réactifs interagissaient de façon plus proximale dans la circulation. Nous concluons donc que la distribution antigénique peut influencer de manière critique la médiation et l'expression morphologique des lésions glomérulaires immunes et qu'elle peut, en partie, rendre compte de variations dans les manifestations cliniques et histologiques de glomérulopathies à médiation par anticorps chez l'homme

Interleukin 1 receptor-associated kinase 1 (IRAK1) mutation is a common, essential driver for Kaposi sarcoma herpesvirus lymphoma

Primary effusion lymphoma (PEL) is an AIDS-defining cancer. It is associated with Kaposi sarcoma-associated herpesvirus. To date, no sequencing studies have been conducted for this cancer. We used X chromosome-targeted next-generation sequencing to identify 33 genes with coding region mutations in 100% of cases, including in interleukin 1 receptor-associated kinase 1 (IRAK1). IRAK1 kinase modulates toll-like receptor signaling-mediated immune signaling. It binds to MyD88 adapter protein, which is mutated in a subset of diffuse large B-cell lymphomas. IRAK1, however, had not been linked to cancer. This IRAK1 mutant is constitutively active and essential for PEL survival. This highlights the importance of innate immunity signaling as drivers for cancer, particularly those caused by viruses. It also suggests IRAK1 kinase may be a potential target for therapy

Labor Productivity - The Use of Staffing Guides and Other Productivity Methods in U.S. Hotels: A Survey Study

This paper examines the issue of labor productivity in hotels. It elaborates on various measurement methods used by American hoteliers including staffing guides, with a special focus on productivity standards. Advantages of physical, financial and mixed methods such as percentage methods, revenue per employee, value-added, Data Envelope Analysis and Stochastic Frontier Analysis are discussed. While the use of percentages and staffing guides were found to be commonplace, results revealed that some fairly stable standards were already in place in the surveyed hotels. Results also revealed that at least in the surveyed companies, few hoteliers attempted to monitor or improve revenue per employee, focusing instead on physical labor inputs and outputs or simple labor percentages

Comparative analysis of slot dimension in lingual bracket systems

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