Dissemination Mechanism Research Articles

ISG15 Is Required for the Dissemination of Vaccinia Virus Extracellular Virions.

Viruses have developed many different strategies to counteract immune responses, and Vaccinia virus (VACV) is one of a kind in this aspect. To ensure an efficient infection, VACV undergoes a complex morphogenetic process resulting in the production of two types of infective virions: intracellular mature virus (MV) and extracellular enveloped virus (EV), whose spread depends on different dissemination mechanisms. MVs disseminate after cell lysis, whereas EVs are released or propelled in actin tails from living cells. Here, we show that ISG15 participates in the control of VACV dissemination. Infection of Isg15-/- mouse embryonic fibroblasts with VACV International Health Department-J (IHD-J) strain resulted in decreased EV production, concomitant with reduced induction of actin tails and the abolition of comet-shaped plaque formation, compared to Isg15+/+ cells. Transmission electron microscopy revealed the accumulation of intracellular virus particles and a decrease in extracellular virus particles in the absence of interferon-stimulated gene 15 (ISG15), a finding consistent with altered virus egress. Immunoblot and quantitative proteomic analysis of sucrose gradient-purified virions from both genotypes reported differences in protein levels and composition of viral proteins present on virions, suggesting an ISG15-mediated control of viral proteome. Lastly, the generation of a recombinant IHD-J expressing V5-tagged ISG15 (IHD-J-ISG15) allowed us to identify several viral proteins as potential ISG15 targets, highlighting the proteins A34 and A36, which are essential for EV formation. Altogether, our results indicate that ISG15 is an important host factor in the regulation of VACV dissemination. IMPORTANCE Viral infections are a constant battle between the virus and the host. While the host's only goal is victory, the main purpose of the virus is to spread and conquer new territories at the expense of the host's resources. Along millions of years of incessant encounters, poxviruses have developed a unique strategy consisting in the production two specialized "troops": intracellular mature virions (MVs) and extracellular virions (EVs). MVs mediate transmission between hosts, and EVs ensure advance on the battlefield mediating the long-range dissemination. The mechanism by which the virus "decides" to shed from the primary site of infection and its significant impact in viral transmission is not yet fully established. Here, we demonstrate that this process is finely regulated by ISG15/ISGylation, an interferon-induced ubiquitin-like protein with broad antiviral activity. Studying the mechanism that viruses use during infection could result in new ways of understanding our perpetual war against disease and how we might win the next great battle.

The Dissemination of Metarhizium brunneum Conidia by Females of the Red Palm Weevil, Rhynchophorus ferrugineus, Suggests a New Mechanism for Prevention Practices.

Direct contact between the conidia of entomopathogenic fungi (EPF) and their host is a prerequisite to successful infection; the host can, therefore, be infected by both direct treatment and by transmission of fungal inoculum from infested surfaces. This unique characteristic makes EPF especially relevant for the control of cryptic insects. In the case of the red palm weevil (RPW) Rhynchophorus ferrugineus, the eggs and larvae are almost inaccessible to direct-contact treatment. The objective of the present study was to investigate the mechanism of conidia transmission from a treated surface to host eggs and larvae. Foam pieces infested with Metarhizium brunneum conidial powder, conidial suspension, or distilled water were used as a laying surface for RPW females. The number of eggs laid was not affected by the EPF treatments and ranged from 2 to 14 eggs per female. However, hatching rate and larval survival were significantly reduced in the conidial powder treatment, resulted in 1.5% hatching and no live larvae. In the conidial suspension treatment, 21% of laid eggs hatched, compared to 72% in the control treatment. In both M. brunneum treatments, females' proboscis, front legs and ovipositor were covered with conidia. The females transferred conidia in both treatments to the laying holes, reaching up to 15 mm in depth. This resulted in reduced egg-hatching rate and significant larval mortality due to fungal infection. The stronger effect on egg and larval survival using dry conidia seemed to result from better conidial adhesion to the female weevil in this formulation. In future studies, this dissemination mechanism will be examined as a prevention strategy in date plantations.

Abstract 2512: TMEM doorways are an actionable target in pancreatic adenocarcinoma

Abstract Introduction: Pancreatic adenocarcinoma (PDAC) is highly lethal due to overwhelming metastatic burden [1]. The mechanisms underpinning metastasis in PDAC have not been identified. The TMEM doorway is the sole portal of intravasation in breast cancer [2, 3]. Importantly, intravasation and dissemination via TMEM doorways is inhibited by Tie2 blockade in breast cancer [4]. We evaluated whether TMEM Doorways are a common targetable mechanism of dissemination in PDAC. Methods: Tumor samples from treatment-naïve patients who underwent curative-intent resection of PDAC were stained for TMEM doorways using a triple immunohistochemistry (IHC) stain (tumor cells expressing the actin regulatory protein Mena, macrophages expressing CD68, and endothelial cells expressing CD31). TMEM doorways were manually counted in the highest TMEM doorway-bearing 400x field (hotspot) identified at 100x scanning magnification for each case. TMEM doorway function was assessed in a murine model of PDAC treated with or without rebastinib (an investigational oral Tie2 inhibitor [4]) by analyzing; (1) TMEM doorway mediated opening using high-molecular weight dextran (155kD-TMR dextran); (2) circulating tumor cells and (3) disseminated tumor cells in murine livers. The Wilcoxon Test was used for the human TMEM doorway scoring comparisons. A student t-test was used for the murine analyses. Results: TMEM doorways were observed in all PDAC tumors from 50 unique treatment-naïve patients. The TMEM doorway score was ≈6x higher than observed in published breast cancer patient samples. Mean TMEM doorway score in “hotspots” from resected human PDAC was 19 and 34 in low- and high-grade tumors, respectively (p-value < 0.01). TMEM doorway mediated opening was decreased by rebastinib treatment (159.8 µm2 ± sd26.5 vs 22.6 µm2 ± sd3.5, p<0.01). PDAC tumor bearing mice treated with rebastinib had a 6.5-fold decrease in the number of circulating tumor cells compared to control (5554 ± sd1017 vs 852 ± sd412 cells/mL blood, p<0.01). PDAC tumor bearing mice treated with rebastinib had fewer disseminated tumor cells compared to untreated mice (11.6 vs 7.0 DTCs/mm2). Conclusions: TMEM doorways appear to be a common mechanism of dissemination in PDAC and breast cancer. TMEM doorway concentration is significantly associated with PDAC tumor grade. Inhibition of Tie2 with the selective inhibitor, rebastinib, preclinically leads to decreased TMEM doorway function and dissemination in PDAC. Tie2 is a promising therapeutic target for decreasing dissemination and potential metastasis in PDAC that warrants further clinical development in the PDAC space. 1.Siegel, R.L., et al. CA: A Cancer Journal for Clinicians, 2022. 72(1): p. 7-33.2.Harney, A.S., et al. Cancer Discovery, 2015. 5(9): p. 932-943.3.Karagiannis, G.S., et al., Science Translational Medicine, 2017. 9(397): p. eaan0026.4.Harney, A.S., et al. Molecular Cancer Therapeutics, 2017. 16(11): p. 2486-2501. Citation Format: Jakeb Petersen, Robert Eddy, Xianjun Ye, Christian Adkisson, David Entenberg, Maja Oktay, John Condeelis, Nicole Panarelli, John Christopher McAuliffe. TMEM doorways are an actionable target in pancreatic adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2512.

Abstract 3354: Isolation and characterization of circulating tumor cells from ductal carcinoma in situ patients using label free technology

Abstract Ductal carcinoma in situ (DCIS) is an early form of breast cancer that has traditionally been characterized by neoplastic cells that are still confined to the mammary ducts. The 20-year breast cancer mortality rate following a DCIS diagnosis, with or without treatment, remains at 3.3%. Despite the overall success in DCIS treatments preventing local invasive breast cancer recurrence, there are still a substantial number of women who, after a DCIS diagnosis, die from metastatic breast cancer. We currently lack predictive tools to identify the high-risk patients who would benefit most from treatment. Emerging clinical and experimental evidence suggest that breast cancer cells may disseminate throughout the body very early in carcinogenesis. These circulating tumor cells (CTCs), rare cancer cells that are shed from the primary tumor into blood, may offer insights into early cancer progression and help identify high-risk patients. Because CTCs can extravasate from blood into tissue and form secondary tumor sites or dormant niches, they are potential biomarkers of metastasis and recurrence. The labyrinth, a high-throughput inertial microfluidic device, has previously demonstrated success in isolating CTCs through inertial separation due to their larger size when compared to white blood cells. Compared to antigen-based isolation technology, inertial microfluidic devices can isolate heterogeneous CTCs with a variety of surface proteins. Using this technology, we isolated and identified CTCs (DAPI+/Pan cytokeratin+/CD45- cells) in 63.2% (12/19) of DCIS patients with an average of 1.27 CTCs per five mls of blood compared to 0.259 CTCs per five mls of blood found in healthy controls (p =0.0078, unpaired t-test). The majority of healthy controls (15/19) had no detectable CTCs. Additionally, immunofluorescence staining showed that 10.5% of CTCs were EpCAM (Epithelial cell adhesion molecule) positive and 26.3% were positive for vimentin, a mesenchymal marker. Single-cell qPCR revealed potential CTCs with no gene expression of traditional white blood cell markers such as CD45, CD20, CD11B, and CD3D. The majority of these cells expressed proliferation (PKM2, TIMP1) and mesenchymal (SPARC, TGFβ1) markers, which can contribute to the cells’ ability to migrate and metastasize. Preliminary targeted DNA sequencing of eight CTC-enriched blood samples identified copy number variations previously associated with higher grade and poorly differentiated DCIS, and single nucleotide variations previously found in DCIS samples (NOTCH1 c.7541_7542del). Ultimately, this study suggests that DCIS generates CTCs prior to microinvasion. CTC isolation and detection may serve as a biomarker for high-risk DCIS and may also be used to explore mechanisms of early cancer dissemination. Citation Format: Brittany Rupp, Brooke Thanasiu, Neha Nagpal, Yan Hong, Dean E. Brenner, Kirsten Tuck, Kirk Herman, Fariba Behbod, Justin Colacino, Max Wicha, Sunitha Nagrath. Isolation and characterization of circulating tumor cells from ductal carcinoma in situ patients using label free technology [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3354.

Primary healthcare needs and service utilisation of people with disability: a data linkage protocol

IntroductionGeneral practitioners (GPs) play a crucial role in the early management and treatment of the comorbidities and complications experienced by people with disability. However, GPs experience multiple constraints, including limited...

‘Shrinking effects’ in cleaner production dissemination – An analysis of the Colombian RedES-CARprogramme

This study evaluates the non-achievement of cleaner production dissemination objectives of firms participating in cleaner production dissemination programmes, often sponsored by public funding. The outcomes help identify, describe, and analyse factors influencing firms’ dropout, resulting in shrinking effects on the number of participating firms benefitting from such mechanisms. The RedES-CAR programme, a significant cleaner production dissemination mechanism in Colombia, served as a source for the empirical data used to test the theoretical model. RedES-CAR fits cleaner production dissemination models in the literature and contains a significant database with information on 490 firms committed to participation in the dissemination mechanisms during 2013–2019. Statistical evaluation identifies how user, organisational, market, and innovation factors influence dropout in diverse phases of the dissemination process. Moreover, the outcomes indicate how factors influencing dropout differ during the dissemination process. Market-based factors are the most relevant in early-phase dissemination for awareness and capacity building, and user and organisational factors influence the assessment, adoption, and multiplication of cleaner production practices. The results highlight the need for managers of dissemination programmes to understand cleaner production dissemination as a multiphase process composed of differentiated dissemination objectives and to raise awareness of the shrinking effect occurring in diverse phases. The societal and environmental benefits of CP dissemination increase with project implementation and multiplication, involving networks of actors. This study contributes to the literature by (i) amplifying the perspective of CP-dissemination literature integrating the lens of diffusion of innovation theory, (ii) offering a conceptual model for operationalising cleaner production dissemination (multiphase), and (iii) structuring factors (negatively) affecting dissemination objectives (multilevel). By doing so, this study also guides the design of strategies and structures to disseminate cleaner production by combining promotional, mitigation, and contingency approaches. The literature has traditionally focused on success stories; this study is an analysis of companies that do not fulfil the dissemination objectives.

BRCA1 and BRCA2 carriers with breast, ovarian and prostate cancer demonstrate a different pattern of metastatic disease compared with non-carriers: results from a rapid autopsy programme.

To catalogue and compare the pattern of metastatic disease in germline BRCA1/2 pathogenic mutation carriers and non-carriers with breast, ovarian and prostate cancer from a rapid autopsy programme. The number of metastases in the major body systems and the proportion of participants with metastases were documented in 50 participants (19 germline mutation carriers). Analysis was conducted on the participants' pattern of disease for the different cancers and mutation subgroups. The four commonly affected organ systems were the digestive (liver only) (82%), respiratory (76%), gastrointestinal (65%) and reticuloendothelial (42%). There were significant differences in the pattern of metastatic breast cancer in BRCA1/2 germline carriers compared with non-carriers. Breast cancer carriers had significantly fewer organ systems involved (median n=3, range=1-3) compared with non-carriers (median n=9, range=1-7) (P=0.03). BRCA1/2 carriers with ovarian carcinomas had significantly more organ systems with metastatic carcinoma (median n=10, range=3-8) than non-carriers (median n=5, range=3-5) (P < 0.001). There were no significant differences in the number of involved systems in BRCA2 carriers compared with non-carriers with prostate cancer (P=1.0). There was an absence of locoregional disease (6.5%) compared with distant disease (93.5%) among the three cancer subtypes (P < 0.001). The majority of metastatic deposits (97%) collected during the autopsy were identified by recent diagnostic imaging. Even though a major limitation of this study is that our numbers are small, especially in the breast cancer carrier group, the metastatic patterns of breast and ovarian cancers may be impacted by BRCA1/2 carrier status, suggesting that tumours derived from patients with these mutations use different mechanisms of dissemination. The findings may focus clinical diagnostic imaging for monitoring metastases where whole-body imaging resources are scant.

The impact of social media marketing on the dissemination of mini program in social network with different community structure

Mini program has become an important infrastructure for the Internetisation of traditional brands. Previous studies have mostly explored the design and development of mini program, but little research has been conducted on the mini program dissemination process. Studying the impact of social media marketing on the mini program dissemination process has important theoretical and applicable value for the in-depth discovery of the mini program dissemination mechanism and the marketing strategies improvement for mini program operators. Based on the classical Susceptible–Infected–Recovered (SIR) epidemic dissemination dynamic model and combined with the Sense, Interest and Interactive, Connect and Diffusion, Action and Share user behaviour consumption model, we propose a mini program propagation model considering social media marketing and community structure, and establish a set of differential equations reflecting the mini program propagation rules. The validity of the model and experimental results are verified through the comparison among the simulation, actual data and numerical experimental results. The experimental results show that the improvement of social media marketing can considerably promote the dissemination of mini program. Surprisingly, with the same social media marketing investment, the dissemination effect of mini program in a weak community is better than that in a strong community. The reason for this is that the higher clustering coefficient fails to offset the negative impact of high modularity on marketing information dissemination. Consequently, compared with the marketing needs of the weak community structure, more social media marketing efforts need to be invested in the strong community structure. Improving social media marketing in the early stages of dissemination can maximise the marketing effect. Moreover, the effect of combining of advertising marketing and interactive marketing in social media marketing is better than that of a single marketing strategy.

Description of Antimicrobial-Resistant Escherichia coli and Their Dissemination Mechanisms on Dairy Farms.

Despite its importance in veterinary medicine, there is little information about antimicrobial resistance (AMR) and its transmission in dairy cattle. The aim of this work is to compare AMR phenotypes and genotypes in resistant Escherichia coli and to determine how the resistance genes spread among the E. coli population on dairy farms in Québec, Canada. From an existing culture collection of E. coli isolated from dairy manure, a convenient selection of the most resistant isolates (a high level of multidrug resistance or resistance to broad-spectrum β-lactams or fluoroquinolones) was analyzed (n = 118). An AMR phenotype profile was obtained for each isolate. Whole genome sequencing was used to determine the presence of resistance genes, point mutations, and mobile genetic elements. In addition, a subset of isolates from 86 farms was taken to investigate the phylogenetic relationship and geographic distribution of the isolates. The average agreement between AMR phenotypes and genotypes was 95%. A third-generation cephalosporin resistance gene (blaCTX-M-15), a resistance gene conferring reduced susceptibility to fluoroquinolones (qnrS1), and an insertion sequence (ISKpn19) were detected in the vicinity of each other on the genome. These genes were harbored in one triplet of clonal isolates from three farms located >100 km apart. Our study reveals the dissemination of resistant E. coli clones between dairy farms. Furthermore, these clones are resistant to broad-spectrum β-lactam and fluoroquinolone antimicrobials.

Global Increase of Antibiotic Resistance Genes in Conjugative Plasmids.

Antibiotic resistance is propagating worldwide, but the predominant dissemination mechanisms are not fully understood. Here, we report that antibiotic resistance gene (ARG) abundance in conjugative plasmids that are recorded in the National Center for Biotechnology Information (NCBI) RefSeq plasmid database is increasing globally, which is likely a key factor in the propagation of resistance. ARG abundance in plasmids increased by 10-fold on a global scale from the year 2000 to the year 2020 (from 0.25 to 2.93 ARG copies/plasmid), with a more pronounced increase being observed in low-to-middle income countries. This increasing trend of plasmid-borne ARGs was corroborated by bootstrap resampling from each year of the NCBI RefSeq plasmid database. The results of a correlation analysis imply that if antibiotic consumption keeps growing at the current rates, a 2.7-fold global increase in the ARG abundance of clinically relevant plasmids may be reached by 2030. High sequence similarities of clinically relevant, conjugative plasmids that are isolated both from clinics and from the environment raise concerns about the environmental resistome serving as a potential ARG maintenance reservoir that facilitates transmission across these ecological boundaries. IMPORTANCE Antibiotic resistance propagation is a significant concern due to its projected impacts on both global health and the economy. However, global propagation mechanisms are not fully understood, including regional and temporal trends in the abundance of resistance plasmids that facilitate antibiotic resistance gene (ARG) dissemination. This unprecedented study reports that ARG abundance in the conjugative plasmids that are recorded in the National Center for Biotechnology Information (NCBI) database and harbor ARGs is increasing globally with antibiotic consumption, especially in low-to-medium income countries. Through network and comparative genomic analyses, we also found high sequence similarities of clinically relevant conjugative resistance plasmids that were isolated from clinical and environmental sources, suggesting transmission between these ecological boundaries. Therefore, this study informs the One Health perspective to develop effective strategies by which to curtail the propagation of plasmid-borne antibiotic resistance.

Genomic characterization of a multidrug-resistant Salmonella Rissen ST469 carrying blaCTX-M-55 gene and Tn6777 isolated from a paediatric patient in China

Here we report a complete genome sequence of a multidrug-resistant Salmonella Rissen, carrying blaCTX-M-55 and Tn6777, isolated from a Chinese paediatric patient. Whole genome of S. Rissen S1905 was sequenced using the Oxford Nanopore MinION and Illumina NovaSeq 6000 platforms. Unicycler was used to perform a de novo assembly of Illumina and Nanopore reads. The genome sequence was annotated using the NCBI Prokaryotic Genome Annotation Pipeline. In silico multilocus sequence typing, plasmid replicons, antimicrobial resistance genes and virulence factors were identified from the genome sequence by multiple bioinformatics tools. Core genome multilocus sequence typing analysis between S. Rissen S1905 and all retrieved from the NCBI GenBank database was performed using BacWGSTdb 2.0 server. Six contigs totaling 5 056 896 bp make up the complete genome sequence of S. Rissen S1905, which includes 1 chromosome and 5 plasmids. The blaCTX-M-55 was embedded in the ISEcp1-blaCTX-M-55-wbuC transposition unit located in an 85 991-bp IncI1 plasmid. However, the pco-sil operon and other eight antimicrobial resistance genes were carried by Tn6777 in the chromosome. There are 162 virulence genes in S1905. S. Rissen S1905 belongs to ST469; the closest relative was another isolate originating from a human faecal specimen in Shanghai, China, which differed by 60 core genome multilocus sequence type alleles. These data on the multidrug-resistant S. Rissen carrying blaCTX-M-55 and Tn6777 can provide a foundation for further studies on the molecular epidemiological characteristics, pathogenicity, antimicrobial resistance mechanisms, and dissemination mechanism of Salmonella.

Baicalein inhibits plasmid-mediated horizontal transmission of the blaKPC multidrug resistance gene from Klebsiella pneumoniae to Escherichia coli.

Carbapenem-resistant bacterial infections pose an urgent threat to public health worldwide. Horizontal transmission of the β-lacatamase Klebsiella pneumoniae carbapenemase (blaKPC) multidrug resistance gene is a major mechanism for global dissemination of carbapenem resistance. Here, we investigated the effects of baicalein, an active ingredient of a Chinese herbal medicine, on plasmid-mediated horizontal transmission of blaKPC from a meropenem-resistant K. pneumoniae strain (JZ2157) to a meropenem-sensitive Escherichia coli strain (E600). Baicalein showed no direct effects on the growth of JZ2157 or E600. Co-cultivation of JZ2157 and E600 caused the spread of meropenem resistance from JZ2157 to E600. Baicalein at 40 and 400 μg/mL significantly inhibited the spread of meropenem resistance. Co-cultivation also resulted in plasmid-mediated transmission of blaKPC from JZ2157 to E600, which was inhibited by baicalein. Therefore, baicalein may be used in clinical practice to prevent or contain outbreaks of carbapenem-resistant infections by inhibiting the horizontal transfer of resistance genes across bacteria species.

The Status and Dilemmas of Chinese Online Fiction in the Global Market

Chinese online novels have gained global influence. After 20 years of rapid development, Chinese web novels have accelerated the pace of "going global", and have demonstrated the cultural self-confidence of the Chinese nation using geographical distribution, genre distribution, and multiple platform distribution from East Asia to North America, gaining a broad space for overseas distribution and activating global cultural and creative energy through Chinese cultural identity. At the same time, the overseas dissemination of Chinese online novels is facing serious piracy, difficulties in protecting rights, homogenization, hollowing out and commercialization of content, difficulties in translation, lack a mature dissemination mechanism, a single business model, and little attention and response to Chinese online novels from the mainstream community abroad. Other difficulties, which require further solutions to promote the better development of overseas dissemination of Chinese online novels.

Going beyond fact-checking to fight health misinformation: A multi-level analysis of the Twitter response to health news stories

Health misinformation has become an unfortunate truism of social media platforms, where lies could spread faster than truth. Despite considerable work devoted to suppressing fake news, health misinformation, including low-quality health news, persists and even increases in recent years. One promising approach to fighting bad information is studying the temporal and sentiment effects of health news stories and how they are discussed and disseminated on social media platforms like Twitter. As part of the effort of searching for innovative ways to fight health misinformation, this study analyzes a dataset of more than 1600 objectively and independently reviewed health news stories published over a 10-year span and nearly 50,000 Twitter posts responding to them. Specifically, it examines the source credibility of health news circulated on Twitter and the temporal, sentiment features of the tweets containing or responding to the health news reports. The results show that health news stories that are rated low by experts are discussed more, persist longer, and produce stronger sentiments than highly rated ones in the tweetosphere. However, the highly rated stories retained a fresh interest in the form of new tweets for a longer period. An in-depth understanding of the characteristics of health news distribution and discussion is the first step toward mitigating the surge of health misinformation. The findings provide insights into understanding the mechanism of health information dissemination on social media and practical implications to fight and mitigate health misinformation on digital media platforms.

Leishmania amazonensis infection impairs VLA-4 clustering and adhesion complex assembly at the adhesion site of J774 cells.

Cutaneous leishmaniasis is an infectious disease that may lead to a single or multiple disseminated cutaneous lesions. The mechanisms involved in Leishmania dissemination to different areas of the skin and the internal organs remain poorly understood. Evidence shows that Very Late Antigen-4 (VLA-4)-dependent phagocyte adhesion is impaired by Leishmania infection, which may be related to the mechanisms of parasite dissemination. We investigated factors potentially associated with decreased VLA-4-mediated adhesion in Leishmania-infected macrophages, including lipid raft-mediated VLA-4 mobilization along the cellular membrane, integrin cluster formation at the cell base (adhesion site), and focal adhesion complex assembly. Phagocytes treated with Methyl-β-Cyclodextrin (MβCD) demonstrated reduced adhesion, similarly to Leishmania amazonensis-infected J774 cells. Infected and MβCD-treated macrophages presented decreased VLA-4 mobilization to the adhesion plane, as well as reduced integrin clustering. Leishmania amazonensis-infected cells exhibited talin depletion, as well as a decreased mobilization of adhesion complex proteins, such as talin and viculin, which were associated with lower VLA-4 concentrations at the adhesion site and limited cell-spreading. Our results suggest that Leishmania infection may modulate the firm adhesion phase of the cell-spreading process, which could contribute to the bloodstream dissemination of infected cells.

Malnutrition-related parasite dissemination from the skin in visceral leishmaniasis is driven by PGE2-mediated amplification of CCR7-related trafficking of infected inflammatory monocytes.

People are infected with Leishmania donovani when the parasite is deposited in the dermis during the blood meal of the sand fly vector. Most infected people develop a subclinical latent infection, but some develop progressive visceral leishmaniasis. Malnutrition is a risk factor for the development of active VL. We previously demonstrated increased parasite dissemination from the skin to visceral organs in a murine model of malnutrition. Here we investigated the mechanism of early parasite dissemination. After delivery of L. donovani to the skin, we found enhanced capture of parasites by inflammatory monocytes and neutrophils in the skin of malnourished mice. However, parasite dissemination in malnourished mice was driven primarily by infected inflammatory monocytes, which showed increased CCR7 expression, greater intrinsic migratory capacity, and increased trafficking from skin to spleen. PGE2 production, which was increased at the site of skin infection, increased monocyte CCR7 expression and promoted CCR7-related monocyte-mediated early parasite dissemination in malnourished mice. Parasite dissemination in monocytes was reduced by inhibition of PGE2, knockdown or silencing of CCR7 in monocytes, and depletion of inflammatory monocytes through administration of diphtheria toxin to CSFR1-DTR transgenic mice that have monocyte-specific DT receptor expression. CCR7-driven trafficking of infected inflammatory monocytes through the lymph node was accompanied by increased expression of its ligands CCL19 and CCL21. These results show that the CCR7/PGE2 axis is responsible for the increased trafficking of L. donovani-infected inflammatory monocytes from the skin to the spleen in the malnourished host. Undernutrition and production of PGE2 are potential targets to reduce the risk of people developing VL. Nutritional interventions that target improved immune function and reduced PGE2 synthesis should be studied in people at risk of developing VL.

Efficient Crash Consistency for NVMe over PCIe and RDMA

This article presents crash-consistent Non-Volatile Memory Express (ccNVMe), a novel extension of the NVMe that defines how host software communicates with the non-volatile memory (e.g., solid-state drive) across a PCI Express bus and RDMA-capable networks with both crash consistency and performance efficiency. Existing storage systems pay a huge tax on crash consistency, and thus cannot fully exploit the multi-queue parallelism and low latency of the NVMe and RDMA interfaces. ccNVMe alleviates this major bottleneck by coupling the crash consistency to the data dissemination. This new idea allows the storage system to achieve crash consistency by taking the free rides of the data dissemination mechanism of NVMe, using only two lightweight memory-mapped I/Os (MMIOs), unlike traditional systems that use complex update protocol and synchronized block I/Os. ccNVMe introduces a series of techniques including transaction-aware MMIO/doorbell and I/O command coalescing to reduce the PCIe traffic as well as to provide atomicity. We present how to build a high-performance and crash-consistent file system named MQFS atop ccNVMe. We experimentally show that MQFS increases the IOPS of RocksDB by 36% and 28% compared to a state-of-the-art file system and Ext4 without journaling, respectively.

Social media &amp; uprisings: The case of the Egyptian revolution in 2011

The present exploratory study adopts the case of the 2011 Egyptian Uprising, as it is particularly illustrative of the climacteric impact of technology on past social and societal conflicts. The latter will be achieved through a systematic review of literature which examines the different ways through which social media was used, as well as the impact it had on facilitating social change with regards to the Egyptian Revolution. The research problem aims to examine the different ways through which social media was used during the Egyptian Uprising, as well as the impact it had on facilitating social change. It caters to a discovery-oriented decision problem and focuses on revealing how social media influences social change while shedding light particularly on the 2011 Egyptian Revolution and the implications social media carried towards it. The conducted exploratory study adequately demonstrates that social media is found to have an undeniable influence on social change, especially as it has been adopted across various phases of the uprising, including its utilization for the initial formation of communities, as well as for promoting the collective movement. Findings of this study will be of great value to future academic studies, as it serves to depict social media as a mechanism of information dissemination in the Arab and/or non-Arab world, whilst correlating to the varied utilities of social media during times of crises and the impact it had on facilitating social change.

Comprehensive Mutation Profiling of Colorectal Cancer Patients With Lung or Liver Metastasis by Targeted Next-Generation Sequencing.

Primary tumor tissue is often analyzed to search for predictive biomarkers and DNA-guided personalized therapies, but there is an incomplete understanding of the discrepancies in the genomic profiles between primary tumors and metastases, such as liver and lung metastases. We performed in-depth targeted next-generation sequencing of 520 key cancer-associated genes for 47 matched primary and metastatic tumor samples which were retrospectively collected. A total of 699 mutations were detected in the 47 samples. The coincidence rate of primary tumors and metastases was 51.8% (n = 362), and compared to patients with liver metastases, patients with lung metastases had a significantly greater coincidence rate (P = .021). The number of specific mutations for the primary tumors and liver and lung metastases was 186 (26.6%), 122 (17.5%), and 29 (4.1%), respectively. Analysis of a patient with all three occurrences, including a primary tumor, liver metastasis, and lung metastasis, indicated a possible polyclonal seeding mechanism for liver metastases. Remarkably, multiple samples from patients with primary and metastatic tumors supported a mechanism of synchronous parallel dissemination from primary tumors to metastatic tumors that were not mediated through pre-metastatic tumors. We also found that the PI3K-Akt signaling pathway significantly altered lung metastases compared to matched primary tumors (P = .001). In addition, patients with mutations in CTCF, PIK3CA, or TP53 and LRP1B, AURKA, FGFR1, ATRX, DNMT3B, or GNAS had larger primary tumor sizes and metastases, especially patients with both LRP1B and AURKA mutations. Interestingly, CRC patients with TP53-disruptive mutations were more likely to have liver metastases (P = .016). In this study, we demonstrate significant differences in the genomic landscapes of colorectal cancer patients based on the site of metastasis. Notably, we observe a larger genomic variation between primary tumors and liver metastasis compared to primary tumors and lung metastasis. These findings can be used for tailoring treatments based on the specific metastatic site.

Green practices of citizens: Sources, stabilization and dissemination (case of Tyumen)

Changing planetary conditions force all actors of social processes to transform social practices. A significant role of society greening belongs to socio-political activism, which becomes a mechanism for launching innovations and changes practices. This article studies the sources of green practices of citizens, the mechanisms of stabilization and dissemination of new practices by questioning the population in Tyumen. To achieve this goal, we formulated five hypotheses. The study did not confirm the hypothesis that green practices arise due to school education, that the authorities are the initiators of greening society, that young people are the driving force of green practices. The obtained data partially confirmed the hypotheses that the green practices of the population are not regular and involvement in green practices occurs through ecological communities. It was found that from 30 to 71% of Tyumen residents know about green practices and are engaged in them. The most common are both practices that have a long history — cleaning the territory, planting trees, and modern practices — taking care of pets, separate waste collection. Respondents consider environmental organizations, activists, and environmental foundations to be initiators and key drivers of green practices. The research contributes to understanding how green practices appear, how they gain their supporters, how they spread in society. Our research can be useful for authorities for improving of the greening process.