Abstract
Lassa fever (LF) survivors develop various clinical manifestations including polyserositis, myalgia, epididymitis, and hearing loss weeks to months after recovery from acute infection. We ...demonstrate a systemic lymphoplasmacytic and histiocytic arteritis and periarteritis in guinea pigs more than 2 months after recovery from acute Lassa virus (LASV) infection. LASV was detected in the arterial tunica media smooth muscle cells by immunohistochemistry, in situ hybridization, and transmission electron microscopy. Our results suggest that the sequelae of LASV infection may be due to virus persistence resulting in systemic vascular damage. These findings shed light on the pathogenesis of LASV sequelae in convalescent human survivors.
Lassa virus persistence in the arterial tunica media smooth muscle cells is associated with systemic lymphoplasmacytic and histiocytic arteritis and periarteritis in convalescent guinea pigs after recovery from acute Lassa fever disease.
Targeted endocytic uptake is a first step toward tissue-specific cytoplasmic macromolecular delivery; however, inefficient escape from the endolysosomal compartment makes this generally impractical ...at present. We report here a targeted cytolysin approach that dramatically potentiates endosomal release of an independently targeted potent gelonin immunotoxin. Fibronectin domains engineered for affinity to EGF receptor or carcinoembryonic antigen were fused to the plant toxin gelonin or bacterial pore-forming cytolysins. These fusion proteins display synergistic activity in both antigen-specific cytotoxicity in vitro, enhancing potency by several orders of magnitude, and in tumor growth inhibition in vivo. In addition, the number of internalized gelonin molecules required to induce apoptosis is reduced from approximately 5 × 10(6) to less than 10(3). Targeted potentiation shows promise for enhancing cytoplasmic delivery of other macromolecular payloads such as DNA, siRNA, and miRNA.
Protein evolution is a critical component of organismal evolution and a valuable method for the generation of useful molecules in the laboratory. Few studies, however, have experimentally ...characterized how fundamental parameters influence protein evolution outcomes over long evolutionary trajectories or multiple replicates. In this work, we applied phage-assisted continuous evolution (PACE) as an experimental platform to study evolving protein populations over hundreds of rounds of evolution. We varied evolutionary conditions as T7 RNA polymerase evolved to recognize the T3 promoter DNA sequence and characterized how specific combinations of both mutation rate and selection stringency reproducibly result in different evolutionary outcomes. We observed significant and dramatic increases in the activity of the evolved RNA polymerase variants on the desired target promoter after selection for 96 h, confirming positive selection occurred under all conditions. We used high-throughput sequencing to quantitatively define convergent genetic solutions, including mutational “signatures” and nonsignature mutations that map to specific regions of protein sequence. These findings illuminate key determinants of evolutionary outcomes, inform the design of future protein evolution experiments, and demonstrate the value of PACE as a method for studying protein evolution.
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IJS, KILJ, NUK, PNG, UL, UM
Hepatocellular carcinoma (HCC) is subject to different management approaches and guidelines according to Eastern and Western therapeutic algorithms. Use of selective internal radiation therapy (SIRT) ...with resin yttrium 90 microspheres for HCC has increased in Asia in recent years, without clearly defined indications for its optimal application. The objective of this systematic review and expert consensus statement is to provide guidance and perspectives on the use of SIRT among patients with HCC in Asia.
A systematic literature review identified current publications on HCC management and SIRT recommendations. A group of 10 experts, representing stakeholder specialties and countries, convened between August 2020 and March 2021 and implemented a modified Delphi consensus approach to develop guidelines and indications for use of SIRT for HCC in Asia. Final recommendations were organized and adjudicated based on the level of evidence and strength of recommendation, per approaches outlined by the American College of Cardiology/American Heart Association and Oxford Centre for Evidence-Based Medicine.
The experts acknowledged a general lack of evidence relating to use of SIRT in Asia and identified as an unmet need the lack of phase 3 randomized trials comparing clinical outcomes and survival following SIRT versus other therapies for HCC. Through an iterative process, the expert group explored areas of clinical relevance and generated 31 guidance statements and a patient management algorithm that achieved consensus.
These recommendations aim to support clinicians in their decision-making and to help them identify and treat patients with HCC using SIRT in Asia. The recommendations also highlight areas in which further clinical trials are needed to define the role of SIRT in management of HCC among Asian populations.
•Management of hepatocellular carcinoma differs between Asia and Western regions.•Comprehensive Asian guidelines on selective internal radiation therapy are needed.•Consensus statements were developed using a 3-step modified Delphi method.•Statements are based on systematic literature review and expert consensus of ≥80%.•Statements are presented for use in treatment of hepatocellular carcinoma in Asia.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
To characterize the contributions of individual amino acids to the structure or function of a protein, researchers have adopted directed evolution approaches, which use iterated cycles of mutagenesis ...and selection or screening to search vast areas of sequence space for sets of mutations that provide insights into the protein of interest.
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IZUM, KILJ, NUK, PILJ, PNG, SAZU, UL, UM, UPUK
Climate change has been associated with a higher frequency of extreme weather events, resulting in an overall increase in morbidity and mortality. Acute otitis media (AOM) is one of the most common ...otolaryngological infections and accounts for 1.5% of emergency department visits. This study aimed to identify associations between extreme weather events and the immediate and delayed risks for AOM-related emergency department visits (EV).
A total of 1,465 AOM-related EVs were identified in the Vienna General Hospital between 2015 and 2018. A distributed lag non-linear model was applied to evaluate the relationship between extreme weather conditions and the total number of AOM-related EVs per day. The relative risk (RR) and cumulative RR (cRR) of single-day events and extended weather events over three days were analyzed over a lag period of 14 days.
AOM-related EVs showed a pronounced seasonality, with the highest occurrence during winter. Single-day weather events affected AOM-related EVs only at high relative humidity. Prolonged extreme weather conditions over three days, however, significantly increased the cRR for AOM-related EVs to 3.15 1.26-7.88;
= 0.014 and 2.14 1.14-4.04;
= 0.018 at mean temperatures of -4°C (1
-percentile - p
) and 0°C (p
) on the same day. Relative humidity of 37% (p
) decreased RR to 0.94 0.88-0.99;
= 0.032 on day 7, while extremely high humidity of 89% (p
) led to an increased cRR of 1.43 1.03-2.00;
= 0.034 on day 7. Heavy prolonged precipitation of 24mm (p
) reduced cRR beginning day 4 up until day 14 to 0.52 0.31-0.86;
= 0.012. Prolonged low atmospheric pressure events of 985hPa (p
) reduced the RR to 0.95 0.91-1.00;
= 0.03, whereas extremely high atmospheric pressure events of 1013hPa (p
) increased the RR to 1.11 1.03-1.20;
= 0.008. Extremely low wind speeds significantly diminished the RR of AOM-related EVs.
While single-day extreme weather events had little impact on the occurrence of AOM-related EVs, extended periods of extreme temperatures, relative humidity, precipitation, wind speeds and atmospheric pressure significantly impacted the RR for AOM-related EVs. These findings could help improve healthcare resource allocation in similar climates and aid in educating patients about the role of environmental factors in AOM.
Abstract Background Context Disc degeneration is associated with the progressive loss of the proteoglycan content of the intervertebral disc (IVD), decreased matrix synthesis, higher concentrations ...of proteolytic enzymes and increased levels of pro-inflammatory cytokines. In previous studies, we have shown that C-C chemokine ligand (CCL)2, CCL3, and CCL5 are highly expressed by cultured nucleus pulposus (NP) and annulus fibrosus (AF) cells that have been treated by interleukin (IL)-1. The major function for these chemokines is to recruit immune cells into the disc. It is unclear if disc cells can respond to these chemokines. Recent studies by Phillips et al. (2015) showed that NP cells express a number of cytokines and chemokine receptors. Purpose The purpose of this study is to determine the gene and protein expression of C-C chemokine receptor (CCR)1, CCR2 and CCR5 in NP and AF cells and to test if these receptors can respond to their ligands in these cells by cell signaling and migration. Study Design/Setting In vitro studies Methods For RNA, surface expression and cell signaling studies, human cells were isolated from the NP and AF tissues collected after spine surgery or from donated spine segments (Gift of Hope Human Donor and Tissue Network of Illinois) and cultured in monolayer. Gene expression of human CCR1, CCR2 and CCR5 was analyzed using real-time PCR. Surface expression of CCR1, CCR2 and CCR5 was analyzed using flow cytometry and fluorescently tagged antibodies specific for these proteins. Extracellular signal-regulated kinase (ERK) phosphorylation was analyzed from the cell lysates of NP and AF cells treated with CCL2 and CCL5 for 1 hour using enzyme-linked immunosorbent assay. Migration of primary rabbit AF cells was assayed using 8 µm Corning Transwell inserts in the presence or absence of CCL5. This study was partially funded by a North American Spine Society 2014 Basic Research Grant Award ($50,000). Results RNA analysis showed that gene expression of CCR1, CCR2 and CCR5 was evident in human NP and AF cells (n=6). Only a small population of NP and AF cells expressed CCR1 (1.9% and 1.2%, respectively) and CCR2 (0.8 and 1.4%, respectively) on the cell surface while a larger percentage expressed CCR5 (12.7% and 11.6%, respectively). Significantly higher levels of ERK phosphorylation were detected in AF cells after treatment with CCL5 and not CCL2. Treatment with either chemokine did not cause significantly higher ERK phosphorylation in NP cells. There was an increase in average AF cell migration in the presence of CCL5. The increase was significant when the migration was induced with CCL5 (500 ng/mL) at both 2 and 6 hour time points. Conclusions CCR5 is expressed at the RNA level and at the cell surface of NP and AF cells. In the presence of CCL5, we detected increased levels of ERK phosphorylation and AF cell migration suggesting that the CCR5 receptors on AF cells are functional. These data suggest that AF cells may have the ability to migrate in response to disc damage or inflammation.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK, ZRSKP
The most commonly reported symptom of post–Ebola virus disease syndrome in survivors is arthralgia, yet involvement of the joints in acute or convalescent Ebola virus infection is not well ...characterized in human patients or animal models. Through immunohistochemistry, we found that the lining synovial intima of the stifle (knee) is a target for acute infection by Ebola virus/Kikwit, Ebola virus/Makona-C05, and Marburg virus/Angola in the rhesus macaque model. Furthermore, histologic analysis, immunohistochemistry, RNAscope in situ hybridization, and transmission electron microscopy showed that synoviocytes of the stifle, shoulder, and hip are a target for mouse-adapted Ebola virus/Yambuku-Mayinga infection during acute disease in rhesus macaques. A time course of infection study with Ebola virus/Kikwit found that the large joint synovium became immunopositive beginning on postinfection day 6. In total, the synovium of 28 of 30 rhesus macaques with terminal filovirus disease had evidence of infection (64 of 96 joints examined). On the basis of immunofluorescence, infected cell types included CD68+ type A (macrophage-like) synoviocytes and CD44+ type B (fibroblast-like) synoviocytes. Cultured primary human fibroblast–like synoviocytes were permissive to infection with Ebola and Marburg viruses in vitro. Because synovial joints include immune privileged sites, these findings are significant for future investigations of filovirus pathogenesis and persistence as well as arthralgias in acute and convalescent filovirus disease.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Human cytomegalovirus (HCMV), a widespread beta-herpes virus, infects a high percentage of gliomas. HCMV is specifically detected in human gliomas at a low level of expression raises the possibility ...that it may regulate the malignant phenotype in a chronic manner. Although HCMV is not recognized as an oncogenic virus, it might dysregulate signaling pathways involved in initiation and promotion of malignancy.Here, our immunohistochemical staining reveals that nucleus staining of the HCMV 86-kDa immediate-early protein (IE86) is markedly increased in GBM (58.56%) compared with that in nontumorous samples (4.20%) and low-grade glioma(19.56%). IE86 staining positively correlates with the staining of activating transcription factor 5 (ATF5) which is essential for glioma cell viability and proliferation suggesting that HCMV IE86 could have important implications in glioma biology. Moreover, we find that the IE86 overexpression enhances glioma cell's growth in vitro and in vivo. We demonstrate that IE86 protein physically interacts with, and acetylates ATF5 thereby promoting glioma cell survival. Therefore, our findings illustrate the biological significance of HCMV infection in accelerating glioma progression, and provide novel evidence that HCMV infection may serve as a therapeutic target in human glioma.
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