Systemic Spread Research Articles

Pharmacokinetic and Environmental Risk Assessment of Prime-2-CoV, a Non-Replicating Orf Virus-Based Vaccine against SARS-CoV-2.

Viral vector vaccines represent a substantial advancement in immunization technology, offering numerous benefits over traditional vaccine modalities. The Orf virus (ORFV) strain D1701-VrV is a particularly promising candidate for vaccine development due to its distinctive attributes, such as a good safety profile, the ability to elicit both humoral and cellular immunity, and its favorable genetic and thermal stability. Despite ORFV's theoretical safety advantages, such as its narrow host range and limited systemic spread post-inoculation, a critical gap persists between these theoretical benefits and the empirical evidence regarding its in vivo safety profile. This discrepancy underscores the need for comprehensive preclinical validations to bridge this knowledge gap, especially considering ORFV's use in humans. Our research introduces Prime-2-CoV, an innovative ORFV-based vaccine candidate against COVID-19, designed to elicit a robust immune response by expressing SARS-CoV-2 Nucleocapsid and Spike proteins. Currently under clinical trials, Prime-2-CoV marks the inaugural application of ORFV in human subjects. Addressing the aforementioned safety concerns, our extensive preclinical evaluation, including an environmental risk assessment (ERA) and detailed pharmacokinetic studies in rats and immunocompromised NOG mice, demonstrates Prime-2-CoV's favorable pharmacokinetic profile, negligible environmental impact, and minimal ERA risks. These findings not only affirm the vaccine's safety and efficacy but also pioneer the use of ORFV-based therapeutics, highlighting its potential for wider therapeutic applications.

The use of DNAPK inhibitors increases the efficacy of Lentiviral vectors and decreases off-target effects.

Abstract Lentiviral vectors are a powerful tool in many models to deliver genetic material to target cells. Many methods of developing gene therapy treatment for multiple human diseases rely on using viral vectors to deliver genetic material into target cells. While viral vectors are a powerful tool, their use is not without its limitations and hurdles. Work must be done to overcome these hurdles to improve the delivery and reliability of vector-based gene therapy mechanisms in a clinically relevant and affordable manner. The primary objective of this study was to enhance the precision and efficacy of lymphocyte modification while minimizing the systemic spread of the vectors. This was achieved through the strategic inhibition of DNA-PK, a key enzyme in DNA repair, to facilitate effective gene integration into lymphocyte genomes. These vectors were administered through trans-splenic injection, a novel approach aimed at directly targeting the lymphocyte-rich environment of the spleen with concurrent DNA-PK inhibition. The results were significant. We observed a markedly high level of gene transduction in splenocytes 30% of T lymphocytes with minimal off-target effects. Notably, there was a substantial reduction in the systemic spread of the vectors, which is a critical factor in mitigating potential side effects. The transduced lymphocytes demonstrated stable gene expression, indicating the success of this method in achieving persistent genetic modification.

PROBLEMS AND PRIORITIES FOR PREVENTING UNDECLARED LABOUR IN UKRAINE

The purpose of the article is to identify the scale of undeclared labour in Ukraine and to study the problems, principles and prospects of its prevention in unstable socio-economic conditions. Methodology. In line with the EU and ILO methodology, the State Labour Service of Ukraine interprets undeclared labour as legal paid activity that is not fully or partially registered in accordance with the law. Results. Undeclared work plays an important role in the practice of business management and competitiveness stimulation in countries with industrial, neo-industrial and post-industrial economies, which has been developed almost since the middle of the 20th century. The paper proves that undeclared work is a broader concept that includes legalised (both official and manipulative) as well as shadowy mechanisms of its occurrence and functioning. Given the problematic performance by the State Labour Service of Ukraine of its inspection and sanctioning powers, as well as the widespread use of non-standard labour relations (with mediation in the performance of employer functions, outsourcing of auxiliary, service and some specialised personnel to companies), tariff-free remuneration systems encourage numerous manipulations in the area of labour declaration and decent wages. The source of problems with the spread of remuneration systems based on an assessment of the complexity of the work performed and the qualifications of employees, as well as with ensuring decent working conditions for agency (loan, temporary) workers, is the unclear legislative regulation of these systems. Practical implications. The priorities for preventing undeclared labour in Ukraine are: a consistent increase in the share of wages in the structure of production costs; strengthening cross-control over the fulfilment of obligations to pay the minimum wage under the tariff-free and hourly wage systems; and the introduction of a sound methodology for harmonising the minimum wage, the subsistence minimum and other basic social standards linked to them. Value/Оriginality. The expediency of identifying institutionalised and manipulative mechanisms of undeclared labour, their formal and illegal bases is confirmed by the classification of the main forms of undeclared labour used by the EU, ILO and the State Labour Service of Ukraine; this approach will increase the complexity and effectiveness of systemic measures to regulate it (control, mitigation, prevention).

Perceptions of general practitioners towards managing dental presentations in Australia: a qualitative study

Background Patients presenting with dental pain are common in general practice, despite dental infections being most appropriately managed with dental treatment to address the cause of the infection. Although antibiotics are not appropriate for the management of localised toothache without signs of systemic spread, general practitioners (GPs) often prescribe antibiotics and analgesics for the management of dental pain. The aim of this study was to explore GPs’ perceptions and management of dental presentations in Australia. Methods Twelve semi-structured interviews were conducted with GPs across Victoria, Australia, between October 2022 and January 2023. Data were thematically analysed. Results The study found that dental pain was the most common presenting complaint for patients attending general practice with a dental problem. Five major themes were identified in this study: knowledge, beliefs about capabilities, emotion, environmental context and resources, and social influences/social professional role and identity. In terms of knowledge and capabilities, GPs would advise patients to seek dental care, as they are aware that antibiotics would not resolve the underlying issue. Challenges for GPs included limited training in oral and dental treatment, as well as emotions, such as patient anxiety and phobia resulting in patients seeking dental care through a GP rather than a dentist. Barriers due to the context, such as access to dental care, long waiting lists within the public dental system and cost, were some of the reasons patients present to general practice rather than a seeing a dentist. Furthermore, issues that influenced GPs’ antibiotic prescribing included patients’ expectations for antibiotics to treat their dental pain, as well as dentists advising their patients to attend their GP for antibiotics prior to dental treatment. Conclusions This study identified factors that influenced GPs’ management of patients with dental conditions. To address these issues, it is imperative to develop interventions addressing patients’ knowledge around oral health, as well as providing improved access to dental care for these patients.

Exploring the Factors Affecting Shared Biking Perception: Insights from Turkey

Understanding consumer perception regarding shared biking services requires the analysis of psychological factors underlying the late diffusion of shared biking services. This approach alone has the potential to develop new business strategies that better respond to users’ needs and facilitate the spread of these mobility systems. By analyzing the factors influencing consumer perception of shared biking services, survey data from 330 respondents from Turkey were used. A structural equation modeling analysis was conducted to investigate consumer perceptions regarding shared biking through the analysis of psychological factors underlying the late diffusion of shared biking services. The statistical tools SPSS version 28 and AMOS version 28 were employed for data analysis. Findings showed that as consumers perceive shared biking services as useful and hedonic, their attitudes become more favorable. Importantly, highlighting the mediating role of compatibility, this study emphasizes the congruence among shared biking services, consumers’ lifestyles, and their traveling habits. Findings also suggest that the perceived complexity has a negative impact on shared biking attitudes. Interestingly, no significant relationship was found between perceived risk and shared biking attitude. The findings provide further support for innovation diffusion and theory of reasoned action, and developed a theoretical framework integrating variables related to both barriers and drivers, thus guiding future studies on the sharing economy. The study also provides insights with the potential to contribute to the development of service design and service marketing strategies that respond better to users’ needs, and also, to facilitate the spread of these mobility systems.

Granulomatous Slack Skin With Lung and Esophagus Involvement: A Case Report and Molecular Analysis.

Granulomatous slack skin (GSS) is a rare subtype of mycosis fungoides, and few cases have been known to spread to the blood, lymph nodes, or viscera. We present a case with early dissemination to the lung. A 27-year-old woman, previously healthy, presented with scattered disseminated scaly patches, associated with vulvar and intergluteal firm swelling and groin-skin induration for 1 year. She also reported mild fatigue and breathlessness on moderate exertion. The patient underwent blood tests, skin biopsies, and computed tomography scan. The skin biopsy showed a mildly atypical T-cell lymphoid infiltrate involving the dermis/hypodermis, with focal epidermotropism, associated with a granulomatous infiltrate and elastophagocytosis. The computed tomography scan revealed bilateral ground-glass lung nodular opacities. Positron emission tomography showed an increased signal in the skin and subcutis around the buttocks, inguinal and mediastinal lymph nodes, and lungs. The lung biopsy confirmed a dense T-cell infiltrate with numerous multinucleated giant cells. Subsequently, esophageal involvement was also observed following biopsy. Molecular analyses demonstrated identical T-cell clones in the skin and lung. After 6 cycles of chemotherapy/localized external radiotherapy, the patient had a partial skin response and stable lung disease. A preferred diagnosis of GSS with systemic spread was made based on clinical/histologic/molecular findings, after considering granulomatous mycosis fungoides and peripheral T-cell lymphoma, not otherwise specified. This case highlights the frequent diagnostic difficulty in distinguishing GSS from an inflammatory granulomatous dermatitis. Pulmonary and esophageal involvements are rare in GSS, and the simultaneous presentation of characteristic cutaneous GSS with systemic disease poses an additional classification challenge.

Effect of low uterine segment involvement on prognosis of early stage endometrial cancer.

Our aim is to investigate the effect of uterine lower segment involvement on prognosis of early-stage endometrial cancer cases diagnosed and treated in our clinic. The file records of 316 cases reviewed retrospectively.Only stage I (a and b, n=209) cases were investigated, because they were more homogeneous group. The lymphovascular invasion rate was found to be higher in patients with stage Ia and uterine lower segment involvement (p < 0.001). Adjuvant treatment requirement was higher in patients with stage Ia and uterine lower segment involvement (p < 0.001). Among stage Ia cases, the recurrence rate between 1 and 3 years was found to be higher in cases with uterine lower segment involvement (p = 0.001). Among the stage Ib cases, lymphovascular invasion was found to be higher in cases with uterine lower segment involvement (p < 0.001). The recurrence rate between 1 and 3 years was found to be higher in stage Ib compared to Ia (p = 0.01). Uterine lower segment involvement was found to be associated with high lymphovascular invasion rate in all stage I cases (p < 0.001). It was determined that the need for adjuvant treatment was higher in cases with uterine lower segment involvement (p < 0.001). It was determined that the probability of recurrence between 1 and 3 years was higher in cases with uterine lower segment involvement (p = 0.007). Uterine lower segment involvement is associated with increased lymphovascular invasion even in the early stages. It is an important risk factor for systemic spread such as lymphovascular invasion, myometrial invasion, and lymph node involvement.

Pathogenic Aspergillus Strains Identification and Antifungal Susceptibility Analysis of 452 Cases with Otomycosis in Jingzhou, China.

To study the distribution of pathogenic Aspergillus strains of otomycosis in central China and the identification of their antifungal sensitivity. We collected external ear canal secretions clinically diagnosed as otomycosis from April 2020 to January 2023 from the Department of Otolaryngology-Head and Neck Surgery in central China. The pathogenic Aspergillus strains were identified through morphological examination and sequencing. The antifungal sensitivity was performed using the broth microdilution method described in the Clinical Laboratory Standard Institute document M38-A3. In the 452 clinical strains isolated from the external ear canal, 284 were identified as Aspergillus terreus (62.83%), 92 as Aspergillus flavus (20.35%), 55 as Aspergillus niger (12.17%). In antifungal susceptibility tests the MIC of Aspergillus strains to bifonazole and clotrimazole was high,all the MIC90 is > 16 ug/mL. However, most Aspergillus isolates show moderate greatly against terbinafine, itraconazole and voriconazole. A. terreus is the most common pathogenic Aspergillus strain in otomycosis in central China. The selected topical antifungal drugs were bifonazole and clotrimazole; the drug resistance rate was approximately 30%. If the infection is persistent and requires systemic treatment, terbinafine and itraconazole can be used. The resistance of Aspergillus in otomycosis to voriconazole should be screened to avoid the systemic spread of infection in immunocompromised people and poor compliance with treatment. However, the pan-azole-resistant strain of Aspergillus should be monitored, particularly in high-risk patients with otomycosis.

УСОВЕРШЕНСТВОВАНИЕ ПРИЕМОВ ПЕРЕВОДА ОЗДОРОВЛЕННЫХ IN VITRO ВИНОГРАДНЫХ РАСТЕНИЙ В УСЛОВИЯ ПОЧВЕННОЙ КУЛЬТУРЫ

The article discusses techniques for improving the properties of a nutrient substrate used for planting healthy in vitro grape plants. The research was carried out on the basis of the laboratory of biotechnology ARRIV&amp;W – branch of FSBSI FRARC on an interspecific hybrid Krasnostop Carpi. The purpuse of the study is to improve the ways of adapting healthy in vitro plants to non–sterile conditions. With microclonal reproduction of grapes, death most often occurs when transferring test tube plants to non-sterile conditions. The selection of the composition of the soil substrate and the improvement of its water-physical and biological properties helps to reduce the risk of plant death. In our studies, peat, garden soil and river sand (1:1:1) were used as part of the substrate, to which “Aquasin hydrogel” and “Trichoderma Veride” endomycorrhizal preparation containing mycelium and spores of Trichoderma viride fungi, strain 471, were added. The plants were planted in a 500 ml container, had been adapted to non-sterile conditions and had been grown for 90 days on racks of accelerated plant cultivation (RAPC). In the process, the indicators of the preservation of initial plants, their growth, development and spread of the root system were noted. As a result, it can be concluded that the combined use of hydrogel and mycorrhiza ensures the best survival and preservation of plants of the Krasnostop Carpi variety. The mycorrhizal preparation promotes better development of the leaf surface, and the addition of hydrogel ensures better development of the root system, which will provide plants with better survival in open ground conditions of the basic mother plantation. Key words: HEALTHY IN VITRO GRAPES, HYDROGEL, MYCORRHIZA, SUBSTRATE, ADAPTATION TO NON-STERILE CONDITIONS

ACCREDITATION AND PROSPECTS OF HACCP SYSTEM IMPLEMENTATION IN FOOD PRODUCTION

The need for accreditation, implementation of the system of analysis of hazardous factors and determination of critical control points (HACCP) at food industry enterprises as a quality management system based on environmental friendliness and product safety is well-founded. The main task in the development of the industry is to increase the competitiveness of products, to strengthen the innovative focus by implementing quality management systems that ensure the quality of products at all stages of its production (life) cycle and contribute to increasing the effectiveness of the work of enterprises [4]. Such a food safety management system, which has proven its effectiveness and is accepted at the international level, is the HACCP system. Guaranteeing food safety is the main goal of applying the HACCP concept to the production process. There are many factors that are not related to the production and processing of products, but have a negative impact on the safety of food products. One aspect of the spread and implementation of this product safety management system is that it is a kind of basis for forming the quality of organic products. When certifying organic products, the necessary prerequisites are documentation of all stages of production and the ability to trace the path "from the doe to the table". Accordingly, the lion's share of the documents that should be at the processing enterprises of organic agricultural products have already been developed, provided that the HACCP system is implemented, and the requirements for product safety are also met. The implementation of the HACCP system requires planning the control of all areas of the technological process, defining the limits of research, application and maintenance of this system. The HACCP plan is drawn up at the enterprise - a document that, in accordance with the principles, determines the procedures and sequence of actions to ensure the control of hazard factors. The HACCP plan covers all areas of product production, from raw material acquisition to product processing or packaging, as well as supply to retail.

Site properties, environmental factors, and crop identify influence soil bacterial communities more than municipal biosolid application

Reducing the environmental impact of Canadian field crop agriculture, including the reliance on conventional synthesised fertilisers, are key societal targets for establishing long-term sustainable practices. Municipal biosolids (MSB) are an abundant, residual organic material, rich in phosphate, nitrogen and other oligo-nutrients, that could be used in conjunction with conventional fertilisers to decrease their use. Though MBS have previously been shown to be an effective fertiliser substitute for different crops, including corn and soybean, there remain key knowledge gaps concerning the impact of MBS on the resident soil bacterial communities in agro-ecosystems. We hypothesised that the MBS fertiliser amendment would not significantly impact the structure or function of the soil bacterial communities, nor contribute to the spread of human pathogenic bacteria, in corn or soybean agricultural systems. In field experiments, fertiliser regimes for both crops were amended with MBS, and compared to corn and soybean plots with standard fertiliser treatments. We repeated this across four different agricultural sites in Quebec, over 2021 and 2022. We sampled MBS-treated, and untreated soils, and identified the composition of the soil bacterial communities via 16S rRNA metabarcoding. We found no indication that the MBS fertiliser amendment altered the structure of the soil bacterial communities, but rather that the soil type and crop identities were the most significant factors in structuring the bacterial communities. Moreover, there was no evidence that the MBS-treated soils were enriched in potential human bacterial pathogens over the two years of our study. Our analysis indicates that not only can MBS function as substitutes for conventional, synthesised fertilisers, but that they also do not disrupt the structure of the resident soil bacterial communities in the short term. Finally, we suggest that the use of MBS in agro-ecosystems poses no greater concern to the public than existing soil bacterial communities. This highlights the significant role MBS could potentially have in reducing the use of conventional industrial fertilisers and improving agricultural production, without risking environmental contamination.

Abstract 3694: Genomic profiling of single cancer cells using the novel single-cell integrated mutational profiling of actionable cancer targets (scIMPACT) assay

Abstract Liquid biopsy provides opportunities to guide therapy decisions using biomarkers such as circulating tumor cells (CTCs) even years after the primary tumor resection. For this, novel methods permitting the genomic profiling of CTCs are needed to characterize systemic cancer spread and identify clinically targetable genomic alterations. Hence, we developed an assay based on the FDA-approved MSK-IMPACT® assay to analyze the genomes of single-cells. We present a series of benchmarking experiments using well-characterized cell lines to demonstrate the performance of our new Single-Cell Integrated Mutation Profiling of Actionable Cancer Targets (scIMPACT) assay.The scIMPACT assay was developed using Ampli1® WGA products of single-cell and pool samples. The workflow consists of an optimized library preparation protocol followed by targeted sequencing with the scIMPACT panel. A bioinformatics mutation calling workflow facilitates the detection of single nucleotide variations (SNVs), indels, and copy number variations (CNVs). To account for WGA associated bias, we developed an in-house proprietary machine learning based classifier algorithm. Two sample collectives were prepared and used for training and validating the workflow. The first collective, including genomic libraries of MDA-MB-453, BT474, BT549 and ZR-75.1 samples, was used for training of the mutation detection workflow. The second collective, consisting of HCC1395/HCC1395BL cell lines representing matched tumor and normal cells, was used to validate the performance of the assay. The scIMPACT assay identified somatic mutations in all WGA products as predicted from bulk gDNA samples and published literature. Notably, evidence of genetic heterogeneity was reported at the single cell level. Pertaining to the SNV detection, the training collective displayed 91% sensitivity and 96% specificity (AUC=0.98) while the validation collective demonstrated similar performance with 84% sensitivity and 96% specificity (AUC=0.90) proving the applicability of our workflow across different datasets. Furthermore, CNV analysis showed high concordance between matching gDNA and WGA samples. Our workflow enables mutational profiling with high specificity and sensitivity and accurate CNV detection in single cells. The scIMPACT assay has been successfully adapted to analyze samples at the single-cell level. Therefore, our method can be applied for the genomic profiling of patient-derived CTCs. Citation Format: Clara Chaiban, Adithi Ravikumar Varadarajan, Jonas Grote, Thomas Ragg, Isabell Blochberger, Vadim Dechand, Jens Warfsmann, Christoph A. Klein, Zbigniew T. Czyż. Genomic profiling of single cancer cells using the novel single-cell integrated mutational profiling of actionable cancer targets (scIMPACT) assay [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3694.

Adoption of the 2A Ribosomal Skip Principle to Track Assembled Virions of Pepper Mild Mottle Virus in Nicotiana benthamiana.

The coat protein (CP) is an important structural protein that plays many functional roles during the viral cycle. In this study, the CP of pepper mild mottle virus (PMMoV) was genetically fused to GFP using the foot-and-mouth disease virus peptide 2A linker peptide and the construct (PMMoV-GFP2A) was shown to be infectious. The systemic spread of the virus was monitored by its fluorescence in infected plants. Electron microscopy and immunocolloidal gold labelling confirmed that PMMoV-GFP2A forms rod-shaped particles on which GFP is displayed. Studies of tissue ultrastructure and virion self-assembly confirmed that PMMoV-GFP2A could be used to monitor the real-time dynamic changes of CP location during virus infection. Aggregations of GFP-tagged virions appeared as fluorescent plaques in confocal laser microscopy. Altogether, PMMoV-GFP2A is a useful tool for studying the spatial and temporal changes of PMMoV CP during viral infection.

γδ T Cells Mediate a Requisite Portion of a Wound Healing Response Triggered by Cutaneous Poxvirus Infection.

Infection at barrier sites, e.g., skin, activates local immune defenses that limit pathogen spread, while preserving tissue integrity. Phenotypically distinct γδ T cell populations reside in skin, where they shape immunity to cutaneous infection prior to onset of an adaptive immune response by conventional αβ CD4+ (TCD4+) and CD8+ (TCD8+) T cells. To examine the mechanisms used by γδ T cells to control cutaneous virus replication and tissue pathology, we examined γδ T cells after infection with vaccinia virus (VACV). Resident γδ T cells expanded and combined with recruited γδ T cells to control pathology after VACV infection. However, γδ T cells did not play a role in control of local virus replication or blockade of systemic virus spread. We identified a unique wound healing signature that has features common to, but also features that antagonize, the sterile cutaneous wound healing response. Tissue repair generally occurs after clearance of a pathogen, but viral wound healing started prior to the peak of virus replication in the skin. γδ T cells contributed to wound healing through induction of multiple cytokines/growth factors required for efficient wound closure. Therefore, γδ T cells modulate the wound healing response following cutaneous virus infection, maintaining skin barrier function to prevent secondary bacterial infection.

Group A streptococcal collagen-like protein 1 restricts tumor growth in murine pancreatic adenocarcinoma and inhibits cancer-promoting neutrophil extracellular traps.

Pancreatic ductal adenocarcinoma (PDAC) is a lethal cancer associated with an immunosuppressive environment. Neutrophil extracellular traps (NETs) were initially described in the context of infection but have more recently been implicated in contributing to the tolerogenic immune response in PDAC. Thus, NETs are an attractive target for new therapeutic strategies. Group A Streptococcus (GAS) has developed defensive strategies to inhibit NETs. In the present work, we propose utilizing intra-tumoral GAS injection to stimulate anti-tumor activity by inhibiting cancer-promoting NETs. Mice harboring Panc02 or KPC subcutaneous tumors injected with three different M-type GAS strains. Tumors and spleens were harvested at the endpoint of the experiments to assess bacterial colonization and systemic spread, while sera were analyzed for humoral responses toward the streptococcal antigens, especially the M1 and Scl1 proteins. Role of the streptococcal collagen-like protein 1 (Scl1) in anti-PDAC activity was assessed in vivo after intratumoral injection with M1 GAS wild-type, an isogenic mutant strain devoid of Scl1, or a complemented mutant strain with restored scl1 expression. In addition, recombinant Scl1 proteins were tested for NET inhibition using in vitro and ex vivo assays assessing NET production and myeloperoxidase activity. Injection of three different M-type GAS strains reduced subcutaneous pancreatic tumor volume compared to control in two different murine PDAC models. Limitation of tumor growth was dependent on Scl1, as isogenic mutant strain devoid of Scl1 did not reduce tumor size. We further show that Scl1 plays a role in localizing GAS to the tumor site, thereby limiting the systemic spread of bacteria and off-target effects. While mice did elicit a humoral immune response to GAS antigens, tested sera were weakly immunogenic toward Scl1 antigen following intra-tumoral treatment with Scl1-expressing GAS. M1 GAS inhibited NET formation when co-cultured with neutrophils while Scl1-devoid mutant strain did not. Recombinant Scl1 protein inhibited NETs ex vivo in a dose-dependent manner by suppressing myeloperoxidase activity. Altogether, we demonstrate that intra-tumoral GAS injections reduce PDAC growth, which is facilitated by Scl1, in part through inhibition of cancer promoting NETs. This work offers a novel strategy by which NETs can be targeted through Scl1 protein and potentiates its use as a cancer therapeutic.

The Fifth Residue of the Coat Protein of Turnip Mosaic Virus Is Responsible for Long-Distance Movement in a Local-Lesion Host and Aphid Transmissibility in a Systemic Host.

HC-Pro and coat protein (CP) genes of a potyvirus facilitate cell-to-cell movement and are involved in the systemic movement of the viruses. The interaction between HC-Pro and CP is mandatory for aphid transmission. Two turnip mosaic virus (TuMV) isolates, RC4 and YC5, were collected from calla lily plants in Taiwan. The virus derived from the infectious clone pYC5 cannot move systemically in Chenopodium quinoa plants and loses aphid transmissibility in Nicotiana benthamiana plants, like the initially isolated virus. Sequence analysis revealed that two amino acids, P5 and A206, of YC5 CP uniquely differ from RC4 and other TuMV strains. Recombination assay and site-directed mutagenesis revealed that the fifth residue of leucine (L) at the N-terminal region of the CP (TuMV-RC4), rather than proline (P) (TuMV-YC5), is critical to permit the systemic spread in C. quinoa plants. Moreover, the single substitution mutant YC5-CPP5L became aphid transmissible, similar to RC4. Fluorescence microscopy revealed that YC5-GFP was restricted in the petioles of inoculated leaves, whereas YC5-CPP5L-GFP translocated through the petioles of inoculated leaves, the main stem, and the petioles of the upper uninoculated leaves of C. quinoa plants. In addition, YC5-GUS was blocked at the basal part of the petiole connecting to the main stem of the inoculated C. quinoa plants, whereas YC5-CPP5L-GFP translocated to the upper leaves. Thus, a single amino acid, the residue L5 at the N-terminal region right before the 6DAG8 motif, is critical for the systemic translocation ability of TuMV in a local lesion host and for aphid transmissibility in a systemic host.

Calculation of product service systems in single and small batch production

AbstractSingle and small batch production is characterized by complex value-added processes and products. The transparent calculation of new offers and change requests is therefore a particular challenge. At the same time, the rising spread of product service systems (PSS) increases the complexity of costing, as additional intangible services have to be calculated precisely. In addition to the challenges posed by such precise calculation of intangible services, companies have to master another complexity driver in the form of PSS. Innovative information and communication technologies (ICT) offer new potential for effective and efficient design of the costing process for the entire life cycle. The rising availability of data along the entire product life cycle significantly increases transparency and, thanks to intelligent analysis algorithms, allows the identification of clear cause-and-effect relationships and forecasting options. The aim of the presented paper is thus to develop a model for calculation of PPS in single and small batch production that exploits the new potential of ICT.

An Italian real-world multicenter study of patients with advanced mixed neuroendocrine non-neuroendocrine neoplasms (MiNENs) of the gastro-entero-pancreatic system treated with chemotherapy.

The aim of this study is to describe the clinical management of an Italian series of patients with advanced gastro-entero-pancreatic (GEP) MiNENs treated in clinical practice. Clinical records of patients from four Italian referral Centers were retrospectively analyzed to correlate clinical/biological data with clinical outcomes. All the surgical specimens were centrally reviewed. Clinical data and surgical samples of 51 patients during 1995-2015 were analyzed. Sites of origin were: 32 colorectal, 14 gastro-esophageal, and 5 pancreatobiliary. Twenty-one out of fifty-one (42.2%) developed metachronous distant metastases. Only 5/51 (9.8%) patients received peri-operative therapy, and 23/51 (45.1%) first-line chemotherapy, mostly fluoropyrimidines/oxaliplatin. The NEN component was poorly differentiated in the whole population. Patients with Ki67 index < 55% in the NEC component had a significantly longer median overall survival (OS) (35.3months; 95% CI 27.1-41.0) than those with Ki67 ≥ 55% (11.9months; 95% CI 9.1-14.0) P = 0.0005. The median OS was 14months (95% CI 10.1-19.1) in the whole cohort, with 11.4months (95% CI 6.2-20.2) in patients who received a first-line therapy. This study confirms that GEP-MiNENs represent a complex disease and that over the past years the clinical management has been predominantly guided by the subjective judgment of the clinicians. Although, in this series, the NEC component appeared mostly responsible for the systemic spread and prognosis on the whole neoplasm, the lack of strong prognostic and predictive factors universally recognized seems to condition their management so far. Future prospective clinical and biomolecular studies could help clinicians to improve clinical management of GEP-MiNENs.

First-in-Class Humanized Antibody against Alternatively Spliced Tissue Factor Augments Anti-Metastatic Efficacy of Chemotherapy in a Preclinical Model of Pancreatic Ductal Adenocarcinoma.

Alternatively spliced tissue factor (asTF) promotes the progression of pancreatic ductal adenocarcinoma (PDAC) by activating β1-integrins on PDAC cell surfaces. hRabMab1, a first-in-class humanized inhibitory anti-asTF antibody we recently developed, can suppress PDAC primary tumor growth as a single agent. Whether hRabMab1 has the potential to suppress metastases in PDAC is unknown. Following in vivo screening of three asTF-proficient human PDAC cell lines, we chose to make use of KRAS G12V-mutant human PDAC cell line PaCa-44, which yields aggressive primary orthotopic tumors with spontaneous spread to PDAC-relevant anatomical sites, along with concomitant severe leukocytosis. The experimental design featured orthotopic tumors formed by luciferase labeled PaCa-44 cells; administration of hRabMab1 alone or in combination with gemcitabine/paclitaxel (gem/PTX); and the assessment of the treatment outcomes on the primary tumor tissue as well as systemic spread. When administered alone, hRabMab1 exhibited poor penetration of tumor tissue; however, hRabMab1 was abundant in tumor tissue when co-administered with gem/PTX, which resulted in a significant decrease in tumor cell proliferation; leukocyte infiltration; and neovascularization. Gem/PTX alone reduced primary tumor volume, but not metastatic spread; only the combination of hRabMab1 and gem/PTX significantly reduced metastatic spread. RNA-seq analysis of primary tumors showed that the addition of hRabMab1 to gem/PTX enhanced the downregulation of tubulin binding and microtubule motor activity. In the liver, hRabMab1 reduced liver metastasis as a single agent. Only the combination of hRabMab1 and gem/PTX eliminated tumor cell-induced leukocytosis. We here demonstrate for the first time that hRabMab1 may help suppress metastasis in PDAC. hRabMab1's ability to improve the efficacy of chemotherapy is significant and warrants further investigation.

Linking enterprise information systems success to female employees’ work–family enrichment in China

Abstract The spread and application of enterprise information systems (EISs) has provided scholars and managers with a new perspective to enhance the work–family enrichment. Based on the work–family enrichment theory, this research aims to examine the ability of female employees to enhance their work–family enrichment by applying the resources accumulated through the use of EISs by combining the technology acceptance model with the DeLone &amp; McLean Information Systems Success Model. The findings based on a survey of 823 full-time female employees in China indicated that the information systems quality factors (including information quality, system quality, and service quality) were positively associated with female employees’ work–family enrichment. In addition, the chained mediating effect of perceived ease of use and perceived usefulness was examined. The results can help female employees to perform positively in both work and family spheres and provide positive support for the promotion of the social fertility policies.