Background/methods: For mechanistic studies, in vitro human B cell differentiation and generation of plasma cells are invaluable techniques. However, the heterogeneity of both T cell-dependent (TD) and T cell-independent (TI) stimuli and the disparity of culture conditions used in existing protocols makes interpretation of results challenging. The aim of the present study was to achieve the most optimal B cell differentiation conditions using isolated CD19⁺ B cells and PBMC cultures. We addressed multiple seeding densities, different durations of culturing and various combinations of TD stimuli and TI stimuli including B cell receptor (BCR) triggering. B cell expansion, proliferation and differentiation was analyzed after 6 and 9 days by measuring B cell proliferation and expansion, plasmablast and plasma cell formation and immunoglobulin (Ig) secretion. In addition, these conditions were extrapolated using cryopreserved cells and differentiation potential was compared. Results: This study demonstrates improved differentiation efficiency after 9 days of culturing for both B cell and PBMC cultures using CD40L and IL-21 as TD stimuli and 6 days for CpG and IL-2 as TI stimuli. We arrived at optimized protocols requiring 2500 and 25.000 B cells per culture well for TD and TI assays, respectively. The results of the PBMC cultures were highly comparable to the B cell cultures, which allows dismissal of additional B cell isolation steps prior to culturing. In these optimized TD conditions, the addition of anti-BCR showed little effect on phenotypic B cell differentiation, however it interferes with Ig secretion measurements. Addition of IL-4 to the TD stimuli showed significantly lower Ig secretion. The addition of BAFF to optimized TI conditions showed enhanced B cell differentiation and Ig secretion in B cell but not in PBMC cultures. With this approach, efficient B cell differentiation and Ig secretion was accomplished when starting from fresh or cryopreserved samples. Conclusion: Our methodology demonstrates optimized TD and TI stimulation protocols for more indepth analysis of B cell differentiation in primary human B cell and PBMC cultures while requiring low amounts of B cells, making them ideally suited for future clinical and research studies on B cell differentiation of patient samples from different cohorts of B cell-mediated diseases.

Cancer-associated fibroblasts (CAF) are highly prevalent cells in the tumor microenvironment in clear cell renal cell carcinoma (ccRCC). CAFs exhibit a pro-tumor effect in vitro and have been implicated in tumor cell proliferation, metastasis, and treatment resistance. Our objective is to analyze the geospatial distribution of CAFs with proliferating and apoptotic tumor cells in the ccRCC tumor microenvironment and determine associations with survival and systemic treatment. Pre-treatment primary tumor samples were collected from 96 patients with metastatic ccRCC. Three adjacent slices were obtained from 2 tumor-core regions of interest (ROI) per patient, and immunohistochemistry (IHC) staining was performed for αSMA, Ki-67, and caspase-3 to detect CAFs, proliferating cells, and apoptotic cells, respectively. H-scores and cellular density were generated for each marker. ROIs were aligned, and spatial point-patterns were generated, which were then used to perform spatial analyses using a normalized Ripley's K function at a radius of 25μm (nK(25)). The survival analyses used an optimal cut-point method, maximizing the log-rank statistic, to stratify the IHC-derived metrics into high and low groups, and multivariable Cox regression analyses were performed accounting for age and International Metastatic RCC Database Consortium (IMDC) risk category. Survival outcomes included overall survival (OS) from the date of diagnosis, OS from the date of immunotherapy initiation (OS-IT), and OS from the date of targeted therapy initiation (OS-TT). Therapy resistance was defined as progression-free survival (PFS) <6 months, and therapy response was defined as PFS >9 months. CAFs exhibited higher cellular clustering with Ki-67+ cells than with caspase-3+ cells (nK(25): Ki-67 1.19; caspase-3 1.05; P = .04). The median nearest neighbor (NN) distance from CAFs to Ki-67+ cells was shorter compared to caspase-3+ cells (15 μm vs 37μm, respectively; P < .001). Multivariable Cox regression analyses demonstrated that both high Ki-67+ density and H-score were associated with worse OS, OS-IT, and OS-TT. Regarding CAFs, only a high H-score was associated with worse OS, OS-IT, and OS-TT. For caspase-3+, high H-score and density were associated with worse OS and OS-TT. Patients whose tumors were resistant to targeted therapy (TT) had higher Ki-67 density and H-scores than those who had TT response. Overall, this ex vivo geospatial analysis of CAF distribution suggests that close proximity clustering of tumor cells and CAFs potentiates tumor cell proliferation, resulting in worse OS and resistance to TT in metastatic ccRCC.

Prothrombotic thrombocytopathy mimicking heparin-induced thrombocytopenia has been observed in patients with severe COVID-19 and after immunisation with the Vaxzevria vaccine. Herein, we discuss the possible pathogenesis of this disorder focusing on the possible involvement of anti-platelet factor 4 (PFA) autoantibodies.

Sickle cell disease is characterized by stiff, “sickled” red blood cells that have difficulty moving through the bloodstream and do not efficiently carry oxygen. It is an inherited disease with severely limited treatment options, and is caused by a point mutation. Its prevalence in black and brown communities makes the already limited treatment options even less accessible. Base editing and prime editing are two relatively recent discoveries in the field of genome editing and were developed after the groundbreaking discovery of the CRISPR Cas9 system. While not fully tested, they hold a lot of promise in providing alternative treatment options for sickle cell disease. Both editing systems are able to install individual point mutations in the beta globin gene, which is where the sickle cell mutation occurs, and can thus cure sickle cell disease (in theory). In this paper we outline the mechanisms of CRISPR-Cas9 systems and base and prime editing, and provide insight into how to apply them to treat SCD. Further investigation should be done on specific editing systems and designs to use to ensure optimal treatment of SCD.

This paper is regarding design and program an Micro-controller Arduino Uno board by using Arduino software to work as a photo-sensor(Active) single axial solar tracker system(SASTS). A solar panel, two photo-resistors (LDR) in two sides (north/south) of the photo-voltaic(PV) and a servo motor are connected to the Uno board, which is running a code that prepared by Arduino software IDE in advanced then it works as a tracking system. Here, the LDRs send the signal of presence or absence of the light to the board and based on that sent signal the Uno reflects a new signal to the servo motor to rotate and finds the light source. Lastly, the photo-sensor single axis tracker is made while Continuously, the system tries to face the panel to the sun and whilst changing the irradiance intensity it starts searching to find the angle of highest irradiance. Based on results that are extracted from the data, the tracker system significantly boosts the output efficiency of the solar panel. By using the Micro-controller Uno board, LDRs, servo motor and special designed mechanical base, the tracking system is constructed, based on acquired data the influence of the STS on the increasing the solar panel efficiency is more obvious. Significantly, the tracker system rises the efficiency of the PV .

With ongoing colony losses driven in part by the Varroa mite and the associated exacerbation of virus load, there is an urgent need to protect honey bees (Apis mellifera) from fatal levels of virus infection and from nontarget effects of insecticides used in agricultural settings. A continuously replicating cell line derived from the honey bee would provide a valuable tool for study of molecular mechanisms of virus – host interaction, for screening of antiviral agents for potential use within the hive, and for assessment of the risk of current and candidate insecticides to the honey bee. However, the establishment of a continuously replicating, honey bee cell line has proved challenging. Here we provide an overview of attempts to establish primary and continuously replicating hymenopteran cell lines, methods (including recent results) for establishing honey bee cell lines, challenges associated with the presence of latent viruses (especially Deformed wing virus), in established cell lines and methods to establish virus-free cell lines. We also describe the potential use of honey bee cell lines in conjunction with infectious clones of honey bee viruses for examination of fundamental virology.

Three-dimensional (3D) cell culture systems can be regarded as suitable platforms to bridge the huge gap between animal studies and two-dimensional (2D) monolayer cell culture to study chronic diseases such as cancer. In particular, the preclinical platforms for multicellular spheroid formation and culture can be regarded as ideal in vitro tumor models. The complex tumor microenvironment such as hypoxic region and necrotic core can be recapitulated in 3D spheroid configuration. Cells aggregated in spheroid structures can better illustrate the performance of anti-cancer drugs as well. Various methods have been proposed so far to create such 3D spheroid aggregations. Both conventional techniques and microfluidic methods can be used for generation of multicellular spheroids. In this review paper, we first discuss various spheroid formation phases. Then, the conventional spheroid formation techniques such as bioreactor flasks, liquid overlay and hanging droplet technique are explained. Next, a particular topic of the hydrogel in spheroid formation and culture is explored. This topic has received less attention in the literature. Hydrogels entail some advantages to the spheroid formation and culture such as size uniformity, the formation of porous spheroids or hetero-spheroids as well as chemosensitivity and invasion assays and protecting from shear stress. Finally, microfluidic methods for spheroid formation and culture are briefly reviewed.

Driven by environmental sustainability concerns, the integration of biobased components in curtain wall systems is gaining traction in both research and the construction market. This paper explores the development and validation of a bio-based façade system within the Basajaun H2020 project. The project aimed to demonstrate the feasibility of introducing environmentally friendly bio-based components into the mature curtain wall façade industry. The paper focuses on identifying technological solutions for replacing key components such as profiles, insulation, and the tightness system with bio-based and less environmental impact alternatives presenting the results achieved in the façade system design of the Basajaun project funded by European Union. These solutions aimed at creating a bio-composite-based curtain wall façade that adheres to current building envelope standards and normative; to implement diverse façade typologies for vision panels, opaque sections, and integrated windows; to engineer the prefabrication process for industrialization, enabling wider market replication and simplified transport and installation. The results demonstrate that the Basajaun façade successfully integrates selected components and meets the performance requirements set by regulations. However, questions remain regarding the workability of bio-based profiles as a commercially viable, ready-to-market solution that can replace traditional aluminum profiles in the curtain wall façade industry.

This review aims to consolidate varied information regarding Port Scanning, and examine the tools, techniques, and detection algorithms used. It explores TCP SYN, Full TCP, UDP, and stealth scans, and shows their functions and abilities. Paired with these techniques, it also shows the tools that they can be used by, such as NMap, Zmap, and MASSCAN. It discusses the various detection techniques such as Signature and Anomaly based detection, and discusses the real world impact of this technology, while commenting on the legal and ethical issues posed.

This study reports the synthesis and characterization of novel per-fluorinated organic polymers with azo- and azomethine-based linkers using nucleophilic aromatic substitution. The polymers reveal variations in the fluorine content via the incorporation of decafluorobiphenyl and hexafluorobenzene linkers, which enhanced their hydrophobic nature. The rich fluorine polymers were slightly soluble in THF and have shown molecular weights between 4886 to 11948 g/mol. All polymers exhibit thermal stability in the range of 350-500°C, with varying thermal stability depending on the fluorine and nitrogen content. Conjugation of the polymers was confirmed through changes in the UV-Vis spectra, with a hypsochromic shift observed in all cases, more pronounced in azo-based fluorinated chains due to H-bonding on the nitrogen sites, chain conformations, and planarity. The optical band gap (Eg) of the polymers was determined from the UV-Vis data, with the Eg values of the azo-based fluorinated polymers being 1 eV higher than those of their corresponding linkers. The cross-linking formation was characterized by porosity measurements, with the azo-based polymer exhibiting the highest surface area of 770 m²/g with a pore volume of 0.35 cm³/g, while the open-chain azomethine-based polymer exhibited the lowest surface area of 285 m²/g with a pore volume of 0.0872 cm³/g. Porous structures with varied hydrophobicities were investigated as adsorbents for separating water-benzene and water-phenol mixtures and selectively binding methane/carbon dioxide gases from the air. The most hydrophobic polymers containing the decafluorbiphenyl linker were suitable for benzene separation, while the best methane uptake values were 6.14 and 3.46 mg/g for DAB-Z-1O and DAB-A-1O, respectively. DAB-Z-1h, with the highest surface area and rich in nitrogen sites, exhibited the highest CO₂ uptake at 298 K (17.25).

G protein-coupled receptors (GPCRs) are amongst the most pharmaceutically relevant and well-studied protein targets, yet unanswered questions in the field leave significant gaps in our understanding of their nuanced structure and function. 3D pharmacophore models are powerful computational tools in silico drug discovery, presenting myriad opportunities for the integration of GPCR structural biology and cheminformatics. This review highlights success stories in the application of 3D pharmacophore modeling to de novo drug design, discovery of biased and allosteric ligands, scaffold hopping, QSAR analysis, hit-to-lead optimization, GPCR de-orphanization, mechanistic understanding of GPCR pharmacology and elucidation of ligand-receptor interactions. Furthermore, advances in the incorporation of dynamics and machine learning will be highlighted. The review will analyze challenges in the field of GPCR drug discovery, detailing how 3D pharmacophore modeling can be used to address them. Finally, we will present opportunities afforded by 3D pharmacophore modeling in the advancement of our understanding and targeting of GPCRs.

Background: Nodal peripheral T-cell lymphomas (nPTCLs) encompass a heterogeneous group of mature and aggressive lymphoid malignancies, including peripheral T-cell lymphoma, not otherwise specified (PTCL/NOS), angioimmunoblastic T-cell lymphoma (AITL) and anaplastic large cell lymphoma (ALCL) ALK-positive and ALK-negative. Their differential diagnosis and prognosis are an issue in the clinical practice. Accurate biomarkers to refine the different subtypes of nPTCLs and to stratify their prognosis are essential to improve their treatment approach. The aim of this study was to test the prognostic impact of GATA3 gene expression, and its capability to discriminate the different subtypes of nPTCLs. Patients and Methods: From 2000 to 2017, 80 patients with nPTCLs were eligible for GATA3 gene expression analysis that was assessed retrospectively by quantitative real time PCR (qRT-PCR) of neoplastic biopsies in Formalin-Fixed Paraffin-Embedded samples (FFPE). Results: Median age was 49 years old (IqR 34-59), 43/80 (53.7%) were male. Median follow-up was 1.72 years. Of them, 36.3% were classified as PTCL/NOS, 31.2% ALK-negative ALCL, 21.2% ALK-positive ALCL and 11.3% AITL. The majority of cases had advanced stage (III/IV). Two-year estimated overall survival (OS) and progression-free survival (PFS) were 52.2% and 39.5%, respectively. The median GATA3 gene expression level was 0.49% (range 0 – 7.07) in all cohort, it was 0.11% for ALK-positive ALCL, 0.46% for ALK-negative ALCL, 0.86% for PTCL/NOS and 0.67% for AITL. The difference of GATA3 gene expression among distinct variants of nPTCLs was statistically significant (p < 0.001). GATA3 gene expression levels ≥ 0.71% discriminate PTCL/NOS from ALK-negative ALCL and AITL with sensitivity of 62% and specificity of 80.3%. GATA3 gene expression levels ≥ median was associated with poor 2-year OS for PTCL/NOS (46.7% x 21.4%, p=0.04) and for ALK-negative ALCL (85.7% x 54.5%, p=0.04). Conclusion: Despite the relative small and heterogeneous group of patients with nPTCLs, GATA3 gene overexpression may be an important biomarker associated with poor prognosis in PTCL/NOS and ALK-negative ALCL. Moreover it may also discriminate different subtypes of nPTCLs. Further studies with larger series of patients should confirm our findings.

Non-melanomatous skin cancers (NMSCs), which include basal and squamous cell carcinoma (BCC and SCC respectively), represent a significant burden on the population as well as an economic load to the health care system, yet treatments of these preventable cancers remain ineffective. Although primary prevention is possible through minimising sunlight exposure, the World Health Organisation estimates that between 2 and 3 million new cases of NMSCs are diagnosed each year, accounting for 1 in 3 of all newly diagnosed cancers. Furthermore, studies have estimated there has been a 2-fold increase in the incidence of NMSCs between 1960s and 1980s. The increase in cases of NMSCs as well as the lack of effective treatments makes the need for novel therapeutic approaches all the more necessary. To rationally develop more targeted treatments for NMSCs, a better understanding of the cell of origin, in addition to the underlying pathophysiological mechanisms that govern the development of these cancers, is urgent. NMSCs are generally thought to arise from specific types of stem cells that become the source of clonal expansion of tumourigenic cells. Previous research on SCC has alluded to these stem cells being localised in the basal compartment of the skin, which ordinarily houses the progenitor cells that contribute towards wound healing and normal cell turnover of overlying epidermal skin layers. More recent research has suggested that commitment to differentiation, which requires exiting the basal, progenitor-cell compartment, is a reversible mechanism. Genetic modifications engage differentiated cells into dedifferentiation, converting them into cancer-initiating cells (CICs) and thereby promoting a tumourigenic environment. Here we explore the most recent developments in the understanding of skin SCC cell of origin, and discuss a case study illustrating the loss of the Grainy-head like 3 (GRHL3) transcription factor in suprabasal layers, which confers a tendency towards tumour development and thereby challenging the “stem cell” theory of tumourigenesis.

: The production of public goods is a necessary condition for the survival of the species but it comes at the expense of individual growth. In a prototype bacterial colony, we model the role of quorum sensing as a resource redistribution mechanism. Two types of bacterial colonies are analyzed, one made up of a single strain and one made up of two different strains. Based on a recent series of experimental data present in the literature, we analyze two types of strains with different extinction times: strains that consume available resources very quickly, therefore becoming extinct quickly, and strains that consume resources slowly and die due to aging. We show that the proposed quorum sensing model describes the main experimental result, namely that coexistence favors the survival of both strains. Furthermore, the production of public goods is maximized when both types of individuals have the maximum proliferation output. Finally, we highlight the role played by so-called dormant cells in the duration of survival time. These cells are of particular interest, because their ability to counteract different types of stress (e.g. the use of antibiotics) still constitutes a challenge.

The generation at the helm faces an unprecedented responsibility in the near future of artificial intelligence. The implications of setting up the founding rules that will regulate the operation of AI are heavy since after they’re set they last forever. Once this first AI is commenced, it can be such that no other subsequent AIs could emerge thereby assuming dominion over its own creation stand. As a result, retaining control becomes necessary. Lest humanity surrender agency to its own creation. At this juncture of big talks, critical issue are raised concerning AI administration owners. Is it appropriate for only a few people to have unrestricted control on AI commands while leaving out all precautionary measure? Therefore, we have to always consider between control and constraint when dealing with AI issues which involves authority plays off against morality. The direction Artificial Intelligence takes in the future depends on the decisions made by today’s generation. We will determine how we are viewed historically in terms of technology based on how well we take on such an important duty. There’s a major turning point ahead of us where we who are the stewards of tomorrow must make a choice that protects humanity’s right to self-determination and also exploits the power of AI for change.

Coronary heart disease (CHD) has become a prevalent issue in Indonesia, resulting in a majority of fatalities and illnesses. The most recent guidelines in the United States for managing patients with chronic coronary syndrome (CCS) have undergone significant changes, with the recommendation for conducting percutaneous coronary intervention (PCI) strictly limited to patients who continue to experience symptoms despite receiving optimal medical treatment (OMT). However, adopting this approach may lead to increased morbidity and mortality due to the presence of vulnerable plaque. Vulnerable plaque is characterized by minimal obstruction during coronary angiography (40-70%) and may not produce ischemia during physiological studies. Identifying vulnerable plaques can be accomplished through several methods, among which computed tomography coronary angiography (CTCA) is the most cost-effective approach. Recently, experts have employed different techniques, either with stents or drug-coated balloons, to manage vulnerable plaques, showing the methods to be safe and effective. The other challenge faced by patients with CCS is the possibility of achieving complete revascularization (CR) to reduce the likelihood of future major adverse cardiac events (MACE) and mortality. Both approaches, identifying and intervening in the vulnerable plaque, are difficult to perform in developing countries due to the limited budget. Studies have shown that plant-based diets (PBD) can reverse atherosclerotic plaque and stabilize vulnerable plaque, possibly leading to CR. PBD is an affordable and effective approach, but it requires healthcare providers skilled in educating and implementing this dietary approach with their patients. Large-scale randomized controlled trials in PBD, specifically aimed at addressing the vulnerable plaque and achieving CR, are necessary before implementing the PBD intervention.

A significant portion of fatalities stemming from acute myocardial infarction (AMI) occur suddenly and without warning. This can be prevented through early detection of coronary stenosis. In recent years, coronary artery calcium (CAC) and computed tomography coronary angiography (CTCA) have been utilized. For instance, a 55-year-old interventional cardiologist had a CAC of 58 with 50% stenosis involving his left anterior descending (LAD) artery. Despite possibly having vulnerable plaque, which carries significant future cardiovascular events, the patient followed a plant-based diet (PBD), took supplements, and continued optimal medical therapy (OMT). After three years, there was a significant regression in both CAC and stenosis. During our discussion, we emphasized the importance of food selection, quantification, and how food is processed (raw) to achieve maximal benefit in regressing atherosclerosis. Additionally, we highlighted the importance of identifying the possibility of vulnerable plaque and the mechanism stabilizing it. The case supported the guidelines set by the American College of Cardiology (ACC) and the American Heart Association (AHA) for managing chronic coronary syndrome (CCS) by emphasizing OMT and lifestyle changes. We hope that in the future, more doctors and patients will take responsibility for implementing lifestyle changes rather than solely relying on medications and sophisticated medical technologies.

In an increasingly complex world there is a real, urgent need for methodologies to enable engineers to model complex sociotechnical systems, as these now seem to describe the majority of systems in use today. This is of course exacerbated by the increasing involvement and augmentation with “black box” AI contributions. Hollnagel produced a methodology, (FRAM) which did allow the analyst in-sights into these systems’ behaviour, but the model-based system engineering applications demand numbers and a quantitative ap-proach. In the last 10 years, this original approach developed to model systems as sets of interactive, interdependent “functions”, (abstracted from agent or component details), has been further developed to the point where it can take the basic data and structures from the current component focussed system engineering “models”, and can pull it all together into dynamic models, (as opposed to static, fixed System Theoretic Process Accimaps), from which analysts can discern how they really work in practice, and predict the emergent behaviours characteristic of complex systems. This paper describes how the FRAM methodology has now been extended to provide these extra, essential attributes. It also describes its implementation using an open-source software, freely available for use and verification on the GitHub site.

As belonging to one of the most isolated continents on our planet, the microbial composition of different environments in Antarctica could hold a plethora of undiscovered species with potential for biotechnological applications. This manuscript delineates our discoveries after an expedition to the Bulgarian Antarctic Base "St. Kliment Ohridski" situated on Livingston Island, Antarctica. Amplicon-based metagenomics targeting the 16S rRNA genes and ITS2 region were employed to assess the metagenomes of the bacterial, fungal and archaeal communities across diverse sites within and proximal to the research station. The predominant bacterial assemblages identified included Oxyphotobacteria, Bacteroidia, Gammaprotobacteria and Alphaprotobacteria. A substantial proportion of cyanobacteria reads were attributed to a singular uncultured taxon within the family Leptolyngbyaceae. The bacterial profile of a lagoon near the base exhibited indications of penguin activity, characterized by a higher abundance of Clostridia, similar to lithotelm samples from Hannah Pt. Although most fungal reads in the samples could not be identified at the species level, noteworthy genera, namely Betamyces and Tetracladium, were identified. Archaeal abundance was negligible, with prevalent groups including Woesearchaeales, Nitrosarchaeum, Candidatus Nitrosopumilus and Marine Group II.

Abstract: Background: Dental fear and uncooperative behavior can hinder dental treatment quality. Pediatric Procedural Sedation and Analgesia (PPSA) is used to facilitate treatment when coping capacity is exceeded. Out-of-hospital PPSA has been associated with more adverse outcomes compared to hospital-based settings. Updated Dutch PPSA guidelines recommending two anesthesiologists and a ventilator during out-of-hospital PPSA have increased costs and raised concerns about the accessibility to specialized high quality dental care for children in the Netherlands. Aim: This study aims to investigate the impact of the updated 2017 guideline on the occurrence rate of adverse events during PPSA, as recorded in the anesthesia complication database of twelve participating Dutch dental clinics, with adjustments made for age, weight, and duration of treatment. Method: Data of 25,872 children treated at 12 dental clinics performing out-of-hospital PPSA between 1997 and 2019 were analyzed. Eligible children were aged 2-18 with ASA classification I and II. A logistic two-level mixed-effects model is used to estimate the updated guideline’s impact on adverse events. Results: In the final model we analyzed the data of 21,759 children in nine dental clinics. The OR of the occurrence rate of an adverse event adjusted for age, weight, and duration of treatment was 0.75 (95% CI 0.64 – 0.89) after the implementation of the updated guideline, compared to before the updated guideline. This outcome was significant with a P=0.001, indicating a protective effect. The average odds per clinic of having an adverse event was 0.04 with a 3.81% chance. Conclusion: Our findings demonstrate a significant reduction in adverse events after the implementation of the updated guideline and highlight the importance of adherence to evidence-based guidelines in improving patient safety and the quality of care provided during pediatric procedural sedation and analgesia in out-of-hospital dental clinics. Our study underlines the necessity of considering the complex underlying mechanisms that play a role in the development and implementation of evidence-based practice and safety initiatives on the occurrence of adverse events.

The healing of osteochondral defects (OCD) resulting from injury, osteochondritis, or osteoarthritis in middle- and old-age individuals, bearing lesions in the cartilage and bone, pain, and loss of joint function, presents challenges to the clinical practitioners because of non-regenerative cartilage and the limitations of current therapies. Bioactive peptide-based osteochondral (OC) tissue regeneration is becoming more popular because it does not have the immunogenicity, misfolding, or denaturation problems associated with original proteins. Periodically, reviews are published on the regeneration of bone and cartilage; however, none of them address the simultaneous healing of these tissues in the complicated heterogeneous environment of the OC interface. As regulators of cell adhesion, proliferation, differentiation, angiogenesis, immunomodulation, and antibacterial activity, potential therapeutic strategies for OC defects utilizing bone and cartilage-specific peptides should be examined and investigated. Therefore, the main goal of this review is to study how they contribute to the healing of OC defects, either alone or in conjunction with other peptides and biomaterials.

In the field of modeling social systems, the use of agent-based models (AM) has become a powerful tool for understanding processes occurring in society, in particular, for studying the spread of infections among the population. The agent model is based on a synthetic population consisting of agents, their interactions and sometimes the territory in which the interaction occurs. The choice of an appropriate method for creating such artificial populations and the selection of parameters becomes a key issue affecting the realism of the model and its ability to reflect real-world scenarios. This article delves into the interplay between agent-based modeling, synthetic population construction, and parameter selection, offering our insight into the complex process of modeling infectious disease dynamics. The review concludes with an exploration of extending the Covasim model [1] to experiments in computational epidemiology. Covasim, a dynamic transmission modeling tool, is known for its ability to adapt to different scenarios and offers predictive information critical for managing epidemiological situations. The Covasim model could include improvements to better simulate real-world epidemic conditions, including a revised transmission strategy to account for accumulation through daily contacts. In addition, we propose to add a module for intercity travel and a visualization package that will significantly expand the functionality of Covasim. These improvements make Covasim a more versatile and efficient tool for modeling and analyzing epidemic processes.

AI-based approaches are increasingly being used to revolutionize vaccine development for COVID-19. Small molecules, peptides, and epitopes are being collected for therapy discovery, which could also direct AI-based models, screening, or generation [1]. AI-based models are being used for drug discovery and vaccine development, and pre-existing data is being leveraged through machine learning approaches for COVID-19 drug discovery and vaccine development [1]. The best candidate targets for future treatment development are being identified and evaluated using AI-based models [1]. AI-based approaches can be used to overcome challenges in manufacturing, storage, logistics, and safety and efficacy issues related to different vaccine candidates for COVID-19 [2]. AI algorithms can help identify the best vaccine candidates for COVID-19 while considering the efficiency of antigenic peptides for immune response generation [2]. The presentation of antigenic peptides by major histocompatibility complex (MHC) class II molecules is the first step after COVID-19 vaccine administration for any vaccine-induced immune response [2]. Thus, AI-based models are being used to identify the best vaccine candidates for COVID-19 and to ensure that the vaccine-induced immune response is efficient and safe. The utilization of AI-based methods to address logistical, manufacturing, storage, safety, and efficacy issues regarding several COVID-19 vaccine candidates will be examined in this study. Additionally, while considering the effectiveness of antigenic peptides for the induction of an immune response, we will determine the best potential targets for the next treatment development and assess how AI-based models can help discover the best vaccine candidates for COVID-19. This research ultimately intends to offer insights into how AI-based techniques can transform COVID-19 vaccine development and how they can be applied to address vaccine development issues. In this paper, we focus on recent advances in using artificial intelligence to develop COVID-19 drugs and vaccines, as well as the potential of intelligent training in discovering COVID-19 therapeutics, highlighting potential challenges and solutions.

The coronavirus disease (COVID-19) is a respiratory tract infection caused by the new virus SARS-CoV-2. In March 2020, the World Health Organization (WHO) declared a worldwide COVID-19 pandemic. Since then, there has been a rush to find preventative or curative treatment. This arrived with the appearance of the COVID-19 vaccine, months after the declaration of the pandemic. In the setting of global COVID-19 immunization, there have been reports of rare skin immune-mediated diseases (IMD) after COVID-19 vaccination, particularly systemic sclerosis. Systemic sclerosis (SSc) is the most reported new onset IMD following messenger RNA (mRNA) COVID-19 vaccination. The pathogenesis of SSc is not fully understood, and the diagnosis is based on clinical symptoms. We report a 54-year-old male, with no prior history of autoimmune disorder, with rapid and progressive skin thickening (Modified Rodnan skin score 30/51), weeks after his mRNA COVID-19 vaccination. Antinuclear antibodies (ANA) showed a strongly positive nuclear fine-speckled pattern, but no SSc autoantibodies were found. The patient was diagnosed with SSc based on the persistence of autoantibodies and the clinical criteria according to the 2013 American College of Rheumatology/European League Against Rheumatism classification. Due to the proximity of the two events, we hypothesized a cross-reaction between COVID-19 vaccination and, in genetically predisposed patients, an emergence of systemic sclerosis. Our case suggests a potential relationship between the mRNA COVID-19 vaccine and new-onset autoimmune diseases. Physicians should be aware of this possible association.

The corrosion resistance of two cast CoCr based alloys, with different amounts of chromium and with different alloying elements in the bulk composition of the alloy, was assessed. Herewith, we investigated the corrosion behavior of Co21Cr8Mo7W and Co29Cr7W by open circuit potential (OCP), potentiodynamic polarization (PP) and electrochemical impedance spectroscopy (EIS) in 0.1 M phosphate buffer solution (PBS) at 37°C for long time immersion. After 1000 hours of immersion, the corrosion current density (icor), estimated from anodic polarization tests, is lower for Co21Cr8Mo7W (i.e. 49 nA cm-2) alloy compared to Co29Cr7W alloy (180 nA cm-2). As regards the corrosion potential (Ecor), a nobler value was observed for Co21Cr8Mo7W (i.e. -59 mV vs. Ag/AgCl) compared to Co29Cr7W (i.e. -114 mV vs. Ag/AgCl). These findings suggest a better corrosion resistance of the film formed on the alloy containing lower amount of Cr and two alloying elements, namely Mo and W. These results are promising ones in terms of medical applications because they open new strategies in obtaining alloys with lower content of chromium and with higher protective properties against corrosion attack.

The global construction sector contributes a significant share of total greenhouse gas (GHG) emissions. In Australia, infrastructure activity alone generates 18% of the GHG emissions budget. The use of low-embodied carbon building materials is crucial to achieving sustainability in the construction sector and to fulfill national and international climate goals. Industrial hemp (Cannabis sativa L.) is considered a promising feedstock for sustainable construction materials because of its biogenic carbon content, fast-growing cycles with low agricultural input requirements, and technical functionality which is comparable to traditional materials. This study has applied the life cycle assessment (LCA) guideline of ISO 14040:2006 to estimate the carbon footprint (CF) expressed in carbon dioxide equivalent (CO2eq) emissions of hemp-based building materials in Western Australia. The functional unit is 1 m2 of hemp-based board, and the system boundary includes cradle-to-gate stages, i.e., pre-farm, on-farm, and post-farm activities. The production of 1 m2 of hemp-based board is estimated to be - 2.302 kgCO2eq. Electricity from the public grid for lignin extraction during the post-farm stage is the main contributor to total CO2eq emissions (26%), followed by urea production (14%) during the pre-farm stage. Overall, the use of electricity from the SWIS during the post-farm stage accounts for 45% of total emissions. Sensitivity analysis shows that the CF of hemp-based boards is highly sensitive to the source of energy, i.e., total replacement of the SWIS by solar power decreases the CF from - 2.30 to -6.07 kgCO2eq (164%). The results suggest that hemp-based boards exhibit lower embodied GHG emissions compared to traditional materials, such as gypsum plasterboards.

TiC particles were prepared from Ti and C powders, and TiC-reinforced Cu-based composites were prepared by powder metallurgy with SPS sintering. The effects of SPS sintering temperature and TiC particle reinforcement phase content on the electrical and mechanical properties of Cu-based composites were investigated. The reinforcement of TiC particles by thermal mismatching at increasing sintering temperature has a significant strengthening effect on the composites, but the high temperature causes softening of the Cu matrix and the growth of grains reduces the properties of the composites. In this study, the effects of porosity and density on the composite properties were not significant.

Background. We created agent-based model for short- and longterm forecasting of COVID-19 and for evaluation how the actions of the regulator affected the human and material resources of the healthcare system. Methods. The model was implemented in the AnyLogic software. It includes two state charts – social network and disease transmission. The COVID-19 Essential Supplies Forecasting Tool (COVID-ESFT, version 2.0) was used to determine healthcare resources needed. Results. Satisfactory results were obtained with long-term (up to 50 days) forecasting in the case of a monotonous change in total cases curve. However, if periods of relative stability are accompanied by sudden outbreaks, relatively satisfactory results were obtained with short-term forecasting, up to 10 days. Simulation of various scenarios showed that the most important place for the spread of infection are families. Wherein the maximum number of cases of COVID-19 is observed in the age group of 26-59 years. Due to a set of measures taken by government agencies, the number of cases in Karaganda city was 3.2 times less than was predicted in “no intervention” scenario. Economic effect is estimated at 40 %. Conclusion. Performed model is an attempt to consider as much as possible the peculiarities of the socio-demographic situation in the country. In the future, we will be prepared to some extent for challenges like those we have experienced in the past three years.

Progress in macrophage research is crucial for numerous applications in medicine, including cancer and infectious diseases. However, the existing methods to manipulate living macrophages are labor intense and inconvenient. Here we show that macrophage membranes can be reconstituted after storage for months at 4C, with their CD206 receptor selectivity and specificity being similar to that in the living cells. Then, we have developed a mannose ligand, specific to CD206, linked with PEG as IR spectroscopy marker to detect binding with the macrophage receptor. PEG was selected due to its unique adsorption band of C-O-C group at IR spectra, which does not overlap with other biomolecule’s spectroscopic feature. Next, competitive binding assay versus the PEG-bound ligand, has enabled selection of other higher-affinity ligands specific to CD206. Further, those higher-affinity ligands were used to differentiate activated macrophages in patient’s bronchoalveolar (BAL) or nasopharyngeal (NPL) lavage. CD206- control cells (HEK293T) showed only non-specific binding. Therefore, biochips based on reconstituted macrophage membranes as well as PEG-trimannoside as an IR spectroscopic marker, can be used to develop new methods facilitating macrophage research and macrophage-focused drug discovery.

Background: The incidence of prostate cancer (PC) has been risen annually. Despite the diagnosis is made mainly with non-metastatic PC, mortality is explained by the metastatic disease (mPC). Without a doubt there is an intermediate scenario in which patients have no mPC but will have initiated a metastatic cascade through an epithelial-mesenchymal transition. There is indeed a need for more and better tools to predict what patients will progress in the future to non-localized clinical disease or already have micrometastatic disease, and therefore, will clinically progress after primary treatment. Biomarkers for predicting mPC are still under development; there are few studies and not much evidence of their usefulness. Summary: This review is focused on tissue-based genomic biomarkers (TBGB) for predicting metastatic disease. We developed four main research questions that will attempt to answer according to the current evidence. Why is important to predict metastatic disease? Which tests are available to predict metastatic disease? What impact should there be on clinical guidelines and clinical practice in predicting metastatic disease? What are current prostate cancer treatments? Key Messages: Knowing useful predict tools could help determine which patients may need multimodal or adjuvant treatment even with a localized disease, and in consequence, what patients do not need more than a single modality of treatment. The importance of predicting metastasis is fundamental, given that once metastasis is diagnosed, the quality of life (QoL) and survival drop dramatically.

Online speech enhancement is of great interest, because of its diverse areas of application. The Demucs architecture has been recently shown to achieve a high level of performance, even with small audio segments, while requiring a relatively small amount of computational resources. However, an issue that it bears, as well as most of state-of-the-art speech enhancement techniques, is that of target speech selection: it is assumed that only one speech source is present. However, in real-life scenarios, more than one speech source may be present, in it is uncertain which speech source within the fed mixture should be enhanced. Thus, it is of interest to complement speech enhancement techniques (such as Demucs-based) with a target selection scheme, so as to ensure that only the source of interest is enhanced. In this work, two target selection strategies are explored: 1) an embedding-based strategy, using a codified sample of the target speech, and 2) a location-based strategy, using a beamforming-based pre-filter to select the target that is in front of a 2-microphone array. It is shown that while both strategies improve the performance of the Demucs-based technique when one or more speech interferences are present, they both have their pros and cons. Specifically, while the beamforming-based strategy achieves overall a better performance compared to the embedding-based strategy, it is sensitive against the location variation of the target speech source which the embedding-based strategy does not suffer from.

The paper explores the potential of a low-cost advanced video-based technique for the assessment of structural damage induced to buildings by seismic loading. A low-cost high-speed video camera was utilized for motion magnification (MM) processing of footages of a two-story reinforced concrete frame building subjected to shaking table tests. The damage after seismic loading was estimated by analyzing the dynamic behavior (i.e. in terms of modal parameters) and the structural deformations of the building in the MM videos. The results by MM were compared for method validation to damage assessment obtained by the analyses of conventional accelerometers and high-precision optical markers tracked by a passive 3D motion capture system. Also, 3D laser scanning to obtain an accurate survey of the building geometry before and after the seismic tests was carried out. In particular, accelerometers were also processed and analyzed by using several stationary and non-stationary techniques with the aim to analyze the linear behavior of the undamaged structure and the nonlinear structural behavior during damaging shaking table tests. The proposed MM-based procedure provided accurate estimate of the main modal frequency and the damage location through the analysis of modal shapes, which were confirmed by advanced analyses of accelerometric data.

The deployment of Electro-Mechanical Actuators plays an important role towards the adoption of the More Electric Aircraft (MEA) philosophy. On the other hand, a seamless substitution of EMAs in place of more traditional hydraulic solutions is still set back due to the shortage of real-life and reliability data regarding their failure modes. One way to work around this problem is providing a capillary EMA Prognostics and Health Management (PHM) system, capable of recognizing failures before they actually undermine the ability of the safety-critical system to perform its functions. The authors have developed a model-based prognostic framework for PMSM based EMAs leveraging a metaheuristic algorithm: Evolutionary (Differential Evolution (DE)) and swarm intelligence (particle swarm (PSO), grey wolf (GWO)) methods are considered. Several failures (dry friction, backlash, short circuit, eccentricity and proportional gain) are simulated thanks to a Reference Model, acting as a Numerical Test Bench, then detected and identified thanks to the envisioned prognostic method, which leverages a low fidelity Monitoring Model. The employed algorithms showed good results and prove that this strategy could be executed in pre-flight checks or during the flight at specific time intervals, with positive impacts on system safety and availability.

Initial choice of Learning Rate is a key part of gradient based methods and has a great effect on the performance of the Deep Learning Model.This paper studies the behavior of multiple gradient based optimization algorithm which are commonly used in Deep Learning and compare their performance on various learning rate. As observed popular choice of optimization algorithms are highly sensitive to various choice of learning rates. Our goal is to find which optimizer has an edge over others for a specific setting. We look at two datasets namely MNIST and CIFAR10 for benchmarking. The results are quite surprising, and it will help us to choose a learning rate more efficiently.

Agent-based models (ABMs) are computational models for simulating the actions and interactions of autonomous agents in time and space. These models allow users to simulate the complex interactions between individual agents and the landscapes they inhabit and are increasingly used in epidemiology to understand complex phenomena and make predictions. However, as the complexity of the simulated systems increases, notably when disease control interventions are considered, model flexibility and processing speed can become limiting. Here we introduce SamPy, an open-source Python library for stochastic agent-based modeling of epidemics. SamPy is a modular toolkit for model development, providing adaptable modules that capture host movement, disease dynamics, and disease control interventions. Memory optimization and design provide high computational efficiency allowing modelling of large, spatially-explicit populations of agents over extensive geographical areas. In this article, we demonstrate the high flexibility and processing speed of this new library. The version of SamPy considered in this paper is available at https://github.com/sampy-project/sampy-paper .

Good estimators are characterized as robust, unbiased, efficient, and consistent. However, the commonly used estimators are weak or lack one or more of these properties. In this article, eight (8) estimators for statistical and geometrical estimations of symmetry/asymmetry, similarity/dissimilarity, identity/unidentity, and feature transformation were proposed following Kabirian-based optinalysis and other operations. The proposed estimators are characterized as invariant (robust) under scaling, location shift, and rotation or reflection. A computing code was written in python language for each of the proposed estimators so that peers can have working codes for application and performance evaluation.

Decision making as a result of system dynamics analysis requires, in practice, a straightforward and systematic modelling capability as well as a high-level of customisation and flexibility to adapt to situations and environments that may vary very much from each other. While in general terms a completely generic approach could be not as effective as ad-hoc solutions, the proper application of modern technology may facilitate agile strategies as a result of a smart combination of qualitative and quantitative aspects. In order to address such a complexity, we propose a knowledge-based approach that integrates the systematic computation of heterogeneous criteria with open semantics. The holistic understanding of the framework is described by a reference architecture and the proof-of-concept prototype developed can support high-level system analysis, as well as it suitable within a number of applications contexts - i.e. as a research/educational tool, communication framework, gamification and participatory modelling. Additionally, the knowledge-based philosophy, developed upon Semantic Web technology, increases the capability in terms of holistic knowledge building and re-use via interoperability. Last but not least, the framework is designed to constantly evolve in the next future, for instance by incorporating more advanced AI-powered features.

There is increasing interest in automating train operations of mainline services, e.g. to increase network capacity. Automatic train operation (ATO) is already achieved by several pilot projects, but not implemented on a large scale. Before the general introduction of new or adapted technologies can have a transformative effect on the operation of such a complex system as train operation on mainlines, they have to pass functional, interoperability and performance tests. A virtual preliminary analysis is one way to ensure a smooth as well as safe introduction and implementation. This paper aims to present an approach that applies to the performance testing of ATO systems. Therefore, methods and test standards for technologies enabling automatic operation in other transport sectors are reviewed. The main findings have been adapted, transformed and combined to be used as a general strategy for virtual performance testing in the railway sector. Specifically, universal performance indicators, namely punctuality, accuracy, energy consumption, safety and comfort, are presented. A layer model for scenario description is adapted from the automotive sector, as well as the definition of different scenario types. Lastly, factors that can influence the performance of an ATO algorithm are identified. To demonstrate the developed approach, a straightforward investigation of a case study is conducted using a microscopic train simulator in combination with an ATO algorithm.

Background: In an age where information is generally accessible, most of the interest these days has focused on how accessible and convenient technology can be. So small and personal, mobile devices can transform our perception of learning by combining both mobility and convenience. Mobile learning is part of the digital learning landscape alongside e-learning and serious games. However, knowledge about effective design of mobile learning experiences remains of interest with a focus on appropriate design models and the embodiments that can be implemented to achieve the intended educational outcomes. Exploring the instructor's perspective on mobile learning is essential. Therefore, the aim of this study was to investigate the Moroccan instructors' perception and practice of mobile learning to inform the development of an ecologically valid mobile learning integration model. Methods: Higher education Instructors (n=41) were recruited to the study. The Moroccan instructors' perception and their experiences regarding their adoption of mobile learning were collected using an online survey. The analysis focused on their mobile use, perceived IT competency, and opinions on mobile learning. Results: We described most of the instructors' considerations regarding integrating mobile technologies into their teaching activities. We found that most of the mobile learning activities defined by the respondents corresponded to relatively advanced use of mobile devices. More promising, instructors have found innovative ways to use the educational potential of mobile devices. However, the prospect of mobile devices was still to challenge. No or poor Wi-Fi connection, number of devices or limited access, sometimes fees or applications incompatibility were identified as reasons and obstacles to mobile learning usage. Conclusion: Mobile learning is mostly perceived positively among Moroccan instructors allowing many applications and usage to enhance teaching and learning. In this study, a better understanding of aspects and factors influencing the integration of mobile learning in the Moroccan educational context is exposed, helping further the development of an ecologically valid mobile learning integration model. Future work on mobile learning should consider the highly paced evolution of mobile technologies, emphasizing the flexibility of integration frameworks to support instructors and learners.

Gender-based violence is considered a serious social and public health problem. Overcoming this situation implies a process that results in the favorable biopsychosocial rehabilitation, the resilient of women. The objective of this study was to analyze the tools, resources and personal and psychosocial mechanisms used by women survivors of gender-based violence. The design was an interpretative phenomenology. It carried out with 22 women who have overcome gender-based violence. Data was collected through personal interviews and narration. The results were grouped into four themes: "Process of violence", "Social resources for coping and overcoming GBV", "Personal tools for coping and overcoming GBV", and "Feelings identified, from the abuse stage to the survival stage". Several studies concluded that overcoming abuse is influenced by the women social network, and it can be the action of these people determining their survival to gender violence. Despite the recognized usefulness of these available resources, it would be desirable to strengthen them in order to be able to drive more women toward survival, assuming a strengthening of coping and overcoming, without forgetting the importance of other support mechanisms such as their family and group therapies.

Malnutrition in children is a global health problem, particularly in developing countries. The effects of an insufficient supply of nutrients on body composition and physiological functions may have implications for drug disposition and ultimately affect the clinical outcome in this vulnerable population. Physiologically based pharmacokinetic (PBPK) modeling can be used to predict the effect of malnutrition as it links physiological changes to pharmacokinetic (PK) consequences. However, the absence of detailed information on body composition and the limited availability of controlled clinical trials in malnourished children complicates the establishment and evaluation of a generic PBPK model in this population. In this manuscript we describe the creation of physiologically-based bridge to a malnourished pediatric population, by combining information on a) the differences in body composition between healthy and malnourished adults and b) the differences in physiology between healthy adults and children. Model performance was confirmed using clinical reference data. This study presents a physiologically-based translational framework for prediction of drug disposition in malnourished children. The model is readily applicable for dose recommendation strategies to address the urgent medicinal needs of this vulnerable population.

This work presents the development and implementation of a distributed navigation system based on computer vision. The autonomous system consists of a wheeled mobile robot with an integrated colour camera. The robot navigates through a laboratory scenario where the track and several traffic signals must be detected and recognized by using the images acquired with its on-board camera. The images are sent to a computer server that processes them and calculates the corresponding speeds of the robot using a cascade of trained classifiers. These speeds are sent back to the robot, which acts to carry out the corresponding manoeuvre. The classifier cascade should be trained before experimentation with two sets of positive and negative images. The number of images in these sets should be considered to limit the training stage time and avoid overtraining the system.

Has anyone ever missed an event because he was confused in days and dates? Do we remember the date of any day without looking at a calendar? Is the current Gregorian Calendar efficient enough for use, and does it facilitate our life or make it more complicated? Have you ever thought about a much simpler way to calculate days and dates in a year? All these questions are answered in this paper, in which the author proposes original optimization algorithm that creates optimal perennial calendars. Results show that there is more than one way to create a perennial calendar, in which the number of days in each month does not change, neither the dates. Hence, all months have the same sequence of days and dates. In other meaning, Monday becomes the first day of every month, and Sunday becomes the last day. Consequently, the calendars become much easier to memorize and very simple to predict the days and dates in any year.

Paper-based sensors, microfluidic platforms and electronic devices have attracted attention in the past couple of decades because they are flexible, can be recycled easily, environmentally friendly, and inexpensive. Here we report a paper aptamer-based potentiometric sensor to detect the whole Zika virus for the first time with a minimum sensitivity of 2.6 nV/Zika and the minimum detectable signal (MDS) of 0.8x1e6 Zika. Our paper sensor works very similar to a P-N junction where a junction is formed between two different wet regions with different electrochemical potentials near each other on the paper. These two regions with slightly different ionic contents, ionic species and concentrations, produce a potential difference given by the Nernst equation. Our paper sensor consisted of a 2-3 mm x 10 mm segments of a paper with a conducting silver paint contact patches on its two ends. The paper is soaked in a buffer solution containing aptamers designed to bind to the capsid proteins on Zika. Atomic force microscopy studies were carried out to show both the aptamer and Zika become immobilized in the paper. We then added the Zika (in its own buffer or simulant Urine) to the region close to one of the silver-paint contacts. The Zika virus (40 nm diameter with 43 kDa or 7.1x10-20 gm weight), became immobilized in the paper’s pores and bonded with the resident aptamers creating a concentration gradient. The potential measured between the two silver paint contacts reproducibly became more negative as upon adding the Zika. We also showed that an LCD powered by the sensor, can be used to detect the sensor output.

Paper-based sensors, microfluidic platforms and electronic devices have attracted attention in the past couple of decades because they are flexible, can be recycled easily, environmentally friendly, and inexpensive. Here we report a paper aptamer-based potentiometric sensor to detect the whole Zika virus for the first time with a minimum sensitivity of 2.6 nV/Zika and the minimum detectable signal (MDS) of 1.2x10⁶ Zika. Our paper sensor works very similar to a P-N junction where a junction is formed between two different wet regions with different electrochemical potentials near each other on the paper. These two regions with slightly different ionic contents, ionic species and concentrations, produce a potential difference given by the Nernst equation. Our paper sensor consisted of a 2-3 mm x 10 mm segments of a paper with a conducting silver paint contact patches on its two ends. The paper is soaked in a buffer solution containing aptamers designed to bind to the capsid proteins on Zika. Atomic force microscopy studies were carried out to show both the aptamer and Zika become immobilized in the paper. We then added the Zika (in its own buffer) to the region close to one of the silver-paint contacts. The Zika virus (40 nm diameter with 43 kDa or 7.1x10^-20 gm weight), became immobilized in the paper’s pores and bonded with the resident aptamers creating a concentration gradient. The potential measured between the two silver paint contacts reproducibly became more negative as upon adding the Zika. We also showed that an LCD powered by the sensor, can be used to detect the sensor output.

Many physiotherapy treatments begin with a diagnosis process. The patient describes symptoms, upon which the physiotherapist decides which tests to perform until a final diagnosis is reached. The relationships between the anatomical components are too complex to keep in mind and the possible actions are abundant. A trainee physiotherapist with little experience naively applies multiple tests to reach the root cause of the symptoms, which is a highly inefficient process. This work proposes to assist students in this challenge by presenting three main contributions: (1) A compilation of the neuromuscular system as components of a system in a Model-Based Diagnosis problem; (2) The PhysIt is an AI-based tool that enables an interactive visualization and diagnosis to assist trainee physiotherapists; and (3) An empirical evaluation that comprehends performance analysis and a user study. The performance analysis is based on evaluation of simulated cases and common scenarios taken from anatomy exams. The user study evaluates the efficacy of the system to assist students in the beginning of the clinical studies. The results show that our system significantly decreases the number of candidate diagnoses, without discarding the correct diagnosis, and that students in their clinical studies find PhysIt helpful in the diagnosis process.

The quadruple burden of diseases, early discharge from hospital and hospital at home have resulted in home-based care services becoming a requirement in South Africa. The home-based care services generate a significant amount of health care risk waste that is mismanaged. However, more attention is given to the health care risk waste generated in hospitals and clinics than to health care risk waste generated by home-based caregivers. Therefore, this study investigates the health care risk waste management practices by home-based caregivers. The study adopted a mixed research approach, qualitative and quantitative methods, using a literature review, interviews, and questionnaires as means of data collection. Results show that there are different types of health care risk waste generated as a result of different activities performed by home-based caregivers, but that the waste was found to be managed in an unsafe manner. The majority of households receiving home-based care did not have basic sanitation facilities such as toilets, running water and waste removal services, aggravating the issue of health care risk waste mismanagement. The study recommends a new policy framework that will lead to safe management practices of generated health care risk waste to be adopted by home-based caregivers.

The application of graph theory in chemical and molecular structure research far exceeds people's expectations, and it has recently grown exponentially. In the molecular graph, atoms are represented by vertices and bonded by edges. Closed forms of multiplicative degree-based topological indices which are numerical parameters of the structure and determine physico-chemical properties of the concerned molecular compound. In this article, we compute and analyze many multiplicative degree-based topological indices of silicon-carbon Si₂C₃-I[p,q] and Si₂C₃-II[p,q].

Recent discovery of triangular boron Nanotubes makes it a competitor of carbon in many respects. Closed forms of M-polynomial of nanotubes produce closed forms of many degree-based topological indices which are numerical parameters of the structure and, in combination, determine properties of the concerned nanotubes. In this report, we give M-polynomials of triangular boron nanotubes and recover many important topological degree-based indices of these nanotubes. We also plot surfaces associated to these nanotubes.

Undernutrition and micronutrient deficiencies such as anemia are considered significant public health challenges in Bangladesh, which enhancing fish consumption is a well-established food-based intervention to address these. This paper documents the establishment of community-based fish chutney production and reports the impact of its consumption on mid-upper arm circumference (MUAC) and hemoglobin (Hb) levels among targeted 150 pregnant and lactating women (PLW) in rural Bangladesh. A fish chutney was developed using locally available ingredients followed by a series of laboratory tests, including nutrient composition, shelf-life and food safety. A community-based fish chutney production process was designed to: 1) supply locally available ingredients for processing; 2) establish two fish drying sites; 3) initiate a community-based production site, and 4) distribute fish chutney to PLW by six women nutrition field facilitators for one year. Then a pre- and post-intervention study was designed for a selected 150 PLW to receive 30g of fish chutney daily for 12 months. Differences in mean mid-upper arm circumference (MUAC) and hemoglobin (Hb) levels pre- and post-consumption were analyzed using one-way analysis of variance. Consumption of 30g of fish-chutney resulted in significant increases of the mean values of Hb levels and MUAC among the targeted PLW.

The fuel cell particularly the Proton Exchange Membrane Fuel Cell (PEMFC) is a new energy technology that plays a critical role in achieving market viability. The PEMFC has high power density, zero CO₂ emissions, and high conversion efficiency. However, with uniform distribution of fuel in PEMFC, we can improve the performance and reduce the cost. The PEMFC will be more efficient and effective in terms of renewable energy as a byproduct only water. Metal foam is used as an alternative for the channels in PEMFC. The model of PEMFC was established by using the ANSYS Fluent 3D Model. Then we proposed four configurations of PEMFC. The outcome demonstrates that Case IV provided outstanding performance if it is compared with the other three cases. In terms of power density, Case IV provided a high power density that is double that of Case I. The porosity of metal foam was investigated. The 0.6 porosity value is found as the optimal value. The uniform distribution of the fuel can be achieved at this value which leads to the outstanding current density.

Separase is a key cysteine protease to separate sister chromatids through digestion of cohesion ring that inhibits chromosome segregation as trigger of metaphase–anaphase transition in eukaryotes and highly regulated its activity by binding with securin and cyclinB-CDK1 complex. Those bindings prevent proteolytic activity of separase until the onset of anaphase. Chromosome missegregation and aneuploidy are frequently observed in malignancies. However, there are some difficulties for biochemical examinations due to instability of separase in vitro and a few spatiotemporal resolution approaches monitoring live separase activity throughout mitotic processes. Here we have developed FRET-based molecular sensors including GFP variants with separase cleavable sequences as donor and covalently attached fluorescent dye as acceptor molecules applicable to conventional live cell imaging and flow cytometric analysis because of efficient live cell uptake. We investigated performance of equivalent molecular sensors localized or not localized inside nucleus under cell cycle control using flow cytometry. Synchronized cell cycle progression rendered significant separase activity detections in both molecular sensors. We obtained consistent outcomes with localized molecular sensor introduction and cell cycle control by fluorescent microscopic observations. We thus established live cell separase activity monitoring systems specifically or statistically that could lead to elucidate separase properties in detail.

The Hippo pathway plays crucial roles in governing various biological processes during tumorigenesis and metastasis. Within this pathway, upstream signaling stimuli activate a core kinase cascade, involving MST1/2 and LATS1/2, which subsequently phosphorylates and inhibits the transcriptional co-activators YAP and its paralog TAZ. This inhibition modulates the transcriptional regulation of downstream target genes, impacting cell proliferation, migration, and death. Despite the acknowledged significance of protein kinases in the Hippo pathway, the regulatory influence of protein phosphatases remains largely unexplored. In this study, we conducted the first gain-of-functional screen for protein tyrosine phosphatases (PTPs) regulating the Hippo pathway. Utilizing a LATS kinase biosensor (LATS-BS), a YAP/TAZ activity reporter (STBS-Luc), and a comprehensive PTP library, we identified numerous novel PTPs that play regulatory roles in the Hippo pathway. Subsequent experiments validated PTPN12, a master regulator of oncogenic receptor tyrosine kinases (RTKs), as a previously unrecognized negative regulator of the Hippo pathway effectors, oncogenic YAP/TAZ, influencing breast cancer cell proliferation and migration. In summary, our findings offer valuable insights into the roles of PTPs in the Hippo signaling pathway, significantly contributing to our understanding of breast cancer bi-ology and potential therapeutic strategies.

Sarcomas are a group of malignancies of mesenchymal origin with plethora of subtypes. Given the sheer heterogeneity of various subtypes and rarity of the disease, management of sarcomas has been challenging with poor patient outcomes. Surgery, radiation therapy and chemotherapy have remained the backbone of treatment in patients with sarcoma. The introduction of immunotherapy has rev-olutionized the treatment of various solid and hematological malignancies. In this review, we discuss basics of immunotherapy and immune microenvironment in sarcomas, various modalities of im-munotherapy such as immune checkpoint blockade, oncolytic viruses, cancer targeted antibodies, vaccine therapy and adoptive cell therapies like CAR T-cell therapy, T cell therapy, TCR therapy.

The human immunodeficiency virus 1 (HIV-1) viral protease (PR) is one of the most studied viral enzymes, and approval of drugs targeting its catalytic activity opened the door to the develop-ment of highly active antiretroviral therapy (HAART). Despite the fact that its crucial role in viri-on maturation is well characterized, an increasing body of research is starting to focus on its abil-ity to cleave host cell proteins, based on recent advances in proteomics and genomics technologies. Such findings are apparently in contrast with the dogma of HIV-1 PR activity being restricted to the interior of nascent virions, and suggest catalytic activity within the host cell environment. Given the limited amount of PR present in the virion at the time of infection, it is tempting to specu-late that such events mainly occur during viral late gene expression, mediated by newly synthe-sized Gag-Pol polyprotein precursors, rather than at a very early stage of infection, before pro-viral integration. Among cellular targets of HIV-1 PR, three major clusters can be identified: pro-teins involved in viral and cellular translation, those controlling cell survival, and restriction fac-tors responsible for innate/intrinsic antiviral responses. Indeed, by cleaving host cell translation initiation factors HIV-1 PR can impair cap-dependent translation, thus promoting IRES-mediated translation of late viral transcripts and viral production, while by targeting several apoptotic fac-tors it modulates cell survival, thus promoting immune evasion and viral dissemination. Addi-tionally, HIV-1 PR counteracts restriction factors incorporated in the virion that would otherwise interfere with nascent virus vitality. Thus, HIV-1 PR appears to modulate host cell function at dif-ferent times and locations during its life cycle, to ensure efficient viral persistency and propaga-tion. This kind of PR-mediated host cell modulation is found in a plethora of different viruses and HIV-1 is no exception, and although we are far from having a complete picture, it is clear that the PR has a multifaceted role in interfering with host machineries to better suit viral replication, and is a field that needs to be explored further.

Cancer stem cells (CSCs) have been identified in many cancer types. This study identified and characterized CSCs in head and neck metastatic malignant melanoma (HNmMM) to regional lymph nodes using induced pluripotent stem cell (iPSC) markers. Immunohistochemical (IHC) staining performed on 20 HNmMM tissue samples demonstrated expression of iPSC markers OCT4, SOX2, KLF4 and c-MYC in all samples while NANOG was expressed at low levels in two samples. Immunofluorescence (IF) staining demonstrated an OCT4+/SOX2+/KLF4+/c-MYC+ CSC subpopulation within the tumor nests (TNs) and another within the peritumoral stroma (PTS) of HNmMM tissues. IF also showed expression of NANOG by some OCT4+/SOX2+/KLF4+/c-MYC+ cells within the TNs in an HNmMM tissue sample that expressed NANOG on IHC staining. In situ hybridization (n=6) and reverse-transcription quantitative polymerase chain reaction (n=5) on the HNmMM samples confirmed expression of all five iPSC markers. Western blotting of four primary cell lines derived from four of the 20 HNmMM tissue samples showed expression of SOX2, KLF4, and c-MYC but not OCT4 and NANOG, and three of these cell lines formed tumorspheres in vitro. We demonstrate the presence of two putative CSC subpopulations within HNmMM, which may be a novel therapeutic target in the treatment of this aggressive cancer.

Cancer cell lines are widely used as in vitro models of tumorigenesis, facilitating fundamental discoveries in cancer biology and translational medicine. Currently, there are few options for glioblastoma (GBM) treatment and limited in vitro models with accurate genomic and transcriptomic characterization. Here, a detailed characterization of a new GBM cell line, namely AHOL1, was conducted in order to fully characterize its molecular composition based on its copy number alteration (CNA) and transcriptome profiling, followed by the validation of key elements associated with GBM tumorigenesis. Large numbers of CNAs and differentially expressed genes (DEGs) were identified. CNAs were distributed throughout the genome, including gains at Xq11.1-q28, Xp22.33-p11.1, Xq21.1-q21.33, 4p15.1-p14, 8q23.2-q23.3 and losses at Yq11.21-q12, Yp11.31-p11.2 and 12p13.31 positions. Nine druggable genes were identified, including HCRTR2, ETV1, PTPRD, PRKX, STS, RPS6KA6, ZFY, USP9Y and KDM5D. By integrating DEGs and CNAs, we identified 57 overlapping genes enriched in fourteen pathways. Altered expression of several cancer-related candidates found in the DEGs-CNA dataset was confirmed by RT-qPCR. Taken together, this first comprehensive genomic and transcriptomic landscape of AHOL1 provides unique resources for further studies and identifies several druggable targets that may be useful for therapeutics and biologic and molecular investigation of GBM.

This paper presents an approach to the application of the Model-Based Systems Engineering (MBSE) and Model-Based Systems Architecting (MBSA) principles to develop a Model-Based Pattern Language (MBPL). It takes too long for systems engineers and architects to develop a new system from scratch, particularly new space-based systems derived from the existing space systems architectures. A pattern language is a holistic view of reusable logical model artifacts; many are interdisciplinary and introductory, if at all. The results are mostly a combination of the application-specific logical solution, which further results in the best possible overall solution. The main benefit of the pattern language is reducing the time and validation required to generate a new space-based system architecture; this approach will develop top-level requirements in the initial phase of the system development. The rationale of the methodology proposed by the paper is as follows, collect, and decompose published literature and other open-source information available on space system architectures and system models; develop SysML models for systems, subsystems, products, assembly, subassembly level, and mission-specific requirements using CAMEO SysML software. Arrange these patterns to develop a functional ontology and construct a logical architecture pattern library. This approach created, updated, and managed SysML pattern language, which evaluated the expedited new model construction. Again, our objective is to develop a logical pattern language using public domain information and evaluate patterns by constructing a new space mission concept—for example, planetary surface habitat.

Cancer is a disease that encompasses multiple and different malignant conditions and is among the leading causes of death in the world. Therefore, the search for new pharmacotherapeutic options and potential candidates that can be used as treatments or adjuvants to control this disease is urgent. Natural products, especially those obtained from plants, have played an important role as a source of specialized metabolites with recognized pharmacological properties against cancer, therefore, they are an excellent alternative to be used. The objective of this research was to evaluate the action of the monoterpene isoespintanol (ISO) against the human tumor cell lines MDA-MB-231, A549, DU145, A2780, A2780-cis and the non-tumor line MRC-5. Experiments with MTT and fluorescence with propidium iodide (PI), DAPI and green plasma revealed the cytotoxicity of ISO against these cells; furthermore, morphological and chromogenic studies revealed the action of ISO on cell morphology and the inhibitory capacity on reproductive viability to form colonies in MDA-MB-231 cells. Likewise, 3D experiments validated the damage in these cells caused by this monoterpene. These results serve as a basis for progress in studies of the mechanisms of action of these compounds and the development of derivatives or synthetic analogues with a better antitumor profile.

Some global difficulties in the modern period include energy storage conversion. Researchers have looked to renewable energy resources to discover long-term answers to this dilemma. Fuel cells are one example of prospective energy producing technology. Although these technologies appear to be very enticing, they are known to be expensive due to high expenses. Catalyst costs, as well as concerns concerning energy density. Pt based Electrocatalysts have traditionally been used to drive ORR and offer efficient power outputs. However, cost constraints have limited the use of these electrocatalysts, rendering them inefficient. Apart from the high price, Pt-based Electrodes are prone to time-dependent drift and CO deactivation, both of which occur gradually. Electrocatalysts deteriorate over time, reducing the efficiency of fuel cells. This is why alternative electrocatalysts are in high demand for future energy applications. This review covers the comparative study of Platinium and Non-Platinium based electrocatalyst in fuel cells, their shortcomings and recent advancement for efficient and optimized electrocatalyst.

The quality of cellular products used in biological research can directly impact the ability to ob-tain accurate results. Epstein-Barr virus (EBV) is a latent virus that spreads extensively world-wide, and cell lines used in experiments may carry EBV and pose an infection risk. The presence of EBV in a single cell line can contaminate other cell lines used in the same laboratory, affecting experimental results. We developed three EBV detection systems: (1) a polymerase chain reac-tion (PCR)-based detection system, (2) a recombinase polymerase amplification (RPA)-based de-tection system, and (3) a combined RPA-lateral flow assay (LFA) detection system. The mini-mum EBV detection limits were 1×103 copy numbers for the RPA-based and RPA-LFA systems and 1×104 copy numbers for the PCR-based system. Both the PCR and RPA detection systems were applied to 192 cell lines, and the results were consistent with those obtained by the EBV assay methods specified in the pharma-ceutical industry standards of the People's Republic of China. 10 EBV-positive cell lines were identified. The combined RPA-LFA system is simple to operate, allowing for rapid results visualization. This system can be implemented in laborato-ries and cell banks as part of a daily quality control strategy to ensure cell quality and experi-mental safety and may represent a potential new technique for the rapid detection of EBV in clinical samples.

This study was conducted to recover valuable metals (silicon and silver) using acid leaching and substitution reactions with copper powder from the pulverized solar cell separated from the PV ribbon. Leaching experiments were conducted according to nitric acid concentration by molar solution as 1M, 2M, and 3M using an ultrasonic cleaner at a reaction temperature of 60 °C for the leaching times of 30 to 120 minutes. Si phases were detected at all leaching reaction times, it was confirmed that the aluminium was removed effectively if the nitric acid leaching time was more than 30 minutes, and silicon with a purity above 99% could be recovered. The silver was precipitated by adding copper, which has greater reducibility than Cu, Al, Si, Pb, and Sn. When the addition of copper powder was 0.75 and 1.0 g, precipitate was produced 0.23 and 0.55 g, respectively. The optimal condition for the recovery of silver by the addition of copper powder was 1 g, where is recovery rate reached 93.3%.

Cancer is the second leading cause of death worldwide. Key to cancer initiation and progression is the crosstalk between cancer cells and their microenvironment. The extracellular matrix (ECM) is a major component of the tumour microenvironment and integrins, main cell-ECM adhesion receptors, are involved in every step of cancer progression. However, accumulating evidence has shown that integrins can act as tumour promoter but also as tumour suppressor factors, revealing that the biological roles of integrins in cancer are complex. This incites a better understating of integrin function in cancer progression. To achieve this goal, simple model organisms, such as Drosophila, offer a great potential to unravel underlying conceptual principles. Here, we find that in the Drosophila wing disc epithelium the PS integrins act as suppressors of tumours induced by a gain of function of the oncogenic form of Ras, RasV12. We show that PS integrin depletion enhances the growth, delamination and invasive behaviour of RasV12 tumour cells, as well as their ability to affect the tumour microenvironment. These results strongly suggest that integrin function as tumour suppressors might be evolutionary conserved. Drosophila can be used to understand the complex tumour modulating activities conferred by integrins, thus facilitating drug development.

Background: Diffuse large B-cell lymphoma is the most common type of non-Hodgkin's lymphoma. Due to the variety of clinical manifestations, accurate diagnosis can be challenging. Here, we present a case of DLBCL that was initially misdiagnosed as a hematoma, highlighting the importance of considering malignancy in the setting of non-responsive soft tissue swelling. Methods: A 76-year-old man presented to the emergency department with right periorbital swelling and ecchymosis. The symptoms first appeared after an injury and did not resolve spontaneously. The patient was taking warfarin for atrial fibrillation. A facial bone CT revealed a large irregular homogeneous mass. An extraconal hematoma was suspected based on clinical history and radiologic findings. Incision and drainage were performed, resulting in evacuation of old blood. Despite continued drainage, the patient's symptoms worsened. A comparison with the previous CT revealed an increase in the size of the lesion. As a result, a surgical excisional biopsy was performed. Results: Biopsy of the mass showed a diffuse infiltrate of lymphocytes around the tissue. The diagnosis of diffuse large B-cell lymphoma was confirmed pathologically. The patient was referred to hematology for further management. Conclusions: DLBCL is a rare disease associated with a poor prognosis. Due to the previous injury and the use of anticoagulants, the patient was initially misdiagnosed as having a hematoma. This delayed the prompt diagnosis and affected the timing of treatment initiation. We must be aware that a malignant mass is a potential cause of unresponsive soft tissue swelling.

Zero-emission trucks for regional and long haul missions are an option for fossil-free freight. The viability of such powertrains and system solutions was studied conceptually in project ESCALATE for trucks with GVW of 40 tonnes and beyond through various prime mover combinations. The study covers battery and fuel cell power sources with different degrees of battery electric as well as H2 and fuel cell operation. As a design basis, two different missions with a single-charge/H2 refill were analysed. The first mission was the VECTO long haul profile repeated up to 750 km, whereas the second was a real 520 km on-road mission in Finland. Based on the simulated energy consumption on the driving cycle, on-board energy demand was estimated, and the initial single-charge operational scenarios were analysed with five different power source topologies. The traction motors of the tractor were dimensioned so that a secondary mission of GVW up to 76 tons on a shorter route can be operated. Based on the powertrain and vehicle model, various infrastructure options for charging and H2 refuelling strategy as well as various operative scenarios with indicative total cost of ownership (TCO) were analysed.

Background – Endobronchial squamous cell carcinoma is one of the most common types of tumors with such a location. Patients often present in advanced stages of the disease, which leads to a targeted therapeutic attitude of pneumonectomy, most of the time. Practicing a lung parenchyma-preserving surgery led us to do this review. Material and Methods - We used three search platforms SCIENCE, MEDLINE and PubMed, in order to identify studies presenting case reports, studies and reviews on endobronchial squamous cell carcinoma. We identified clinical and paraclinical features of endobronchial squamous cell carcinoma. All selected articles were in English and contained clinical criteria of endobronchial squamous cell carcinoma, autofluorescence bronchoscopy in endobronchial squamous cell carcinoma, imaging features of endobronchial squamous cell carcinoma, blood tumor markers specific for lung squamous cell carcinoma, histopathological features of endobronchial squamous cell carcinoma. Results - 73 articles were analyzed, from which 47 articles were selected as bibliographic references. We have presented the criteria found for endobronchial squamous cell carcinoma in order to highlight the main characteristics and high reliability technologies for the detection of this type of cancer. Conclusions – The current literature review highlights the clinical and paraclinical characteristics of endobronchial squamous cell carcinoma. It aims to open new horizons for research and early detection, frequent practice of lung parenchymal preservation surgery.

Osteitis fibrosa cystica (OFC) and Brown Tumour are two related but distinct types of bone lesions that result from overactivity of osteoclasts most often associated with chronic kidney disease (CKD). Despite their potential consequences, these conditions are poorly understood because of their rare prevalence and variability in their clinical manifestation. Canonically, OFC and Brown Tumours are caused by secondary hyperparathyroidism in CKD. Recent literature showed that multiple factors such as hyperactivation of the renin-angiotensin-aldosterone system and chronic inflammation may also contribute to the occurrence of these diseases through osteoclast activation. Moreover, hotspot KRAS mutations were identified in these lesions placing them in the spectrum of RAS-MAPK-driven neoplasms, while until recently thought to be reactive lesions. Some risk factors contributed to the occurrence of OFC and Brown Tumour such as age, gender, comorbidities, and certain medications. The diagnosis of OFC and Brown Tumour includes clinical symptoms involving chronic bone pain and laboratory finding of hyperparathyroidism. In radiological imaging, the X-ray and Computed tomography (CT) scan could show lytic or multi-lobular cystic alterations. Histologically both lesions are characterized by clustered osteoclast in a fibrotic hemorrhagic background. Based on the latest understanding of the mechanism of OFC, this review elaborates on the manifestation, diagnosis, and available therapies that can be leveraged to prevent the occurrence of OFC and Brown Tumour.

IQGAP1 is a multi-functional scaffold protein. However, its role in B cell development and function is not known. Here, we demonstrate Iqgap1mice contained significantly increased numbers of B220+ B, B220+IgM pro/pre-B, and B220+IgM+ immature-B cells in the bone marrow. Here, we show IQGAP1 as the essential scaffold that regulates early B cell development and function. Iqgap1mice contained significantly increased numbers of B220+ B, B220+IgM pro/pre-B, and B220+IgM+ immature-B cells in the bone marrow. New-forming and follicular B cell numbers were increased, while the marginal zone B cell numbers were significantly reduced in the spleen of the Iqgap1mice. Lack of IQGAP1 reduced T-dependent and T-independent humoral responses. Mechanistically, the lack of IQGAP1 considerably decreased the phosphorylation of Mek1/2, Erk1/2, and Jnk1/2. B cells from Iqgap1mice failed to suppress IL-7R-mediated activation of Stat5a/b, an essential step for cell-cycle exit and initiate light-chain recombination, leading to a reduction in RS rearrangement frequency. Lack of IQGAP1 resulted in a significant reduction of IRF4 and IRF8 in BM B cells. Our study provides the first evidence that IQGAP1-based signalosome is essential for the development and functions of B cells.

The Zika virus (ZIKV) epidemic brought new discoveries regarding arboviruses, especially flaviviruses, as ZIKV was described as sexually and vertically transmitted. The latter shows severe consequences for the embryo/fetus, such as congenital microcephaly and deficiency of the neural system, currently known as Congenital ZIKV Syndrome (CZS). To better understand ZIKV dynamics in trophoblastic cells present in the first trimester of pregnancy (BeWo, HTR-8, and control cell HuH-7), an experiment of viral kinetics was performed for African MR766 low passage and Asian-Brazilian IEC ZIKV lineages. The results were described independently, and demonstrated that the three placental cells lines are permissive and susceptible to ZIKV. We noticed cytopathic effects that are typical in vitro viral infection in BeWo and HTR-8. Regarding kinetics, MR766lp showed peaks of viral loads in 24 and 48 hpi for all cell types tested, as well as marked cells death after peak production. On the other hand, HTR-8 lineage inoculated with ZIKV-IEC exhibited increased viral production in 144 hpi, with a peak between 24 and 96 hpi. Furthermore, IEC had peak variations of viral production for BeWo in 144 hpi. Both cells types continued alive during the process of viral replication. Considering such in vitro results, the hypothesis that maternal-fetal transmission is probably a way of virus transmission between the mother and the embryo/fetus is maintained.

Sperm gain fertilization ability and forward movement characteristics during epididymal transport. Sperm maturation is inseparable from the processing of the epididymis. Compared with other reproductive organs, relatively little is known about the epididymis. However, with the development of molecular biology and the improvement of single-cell sequence technology, we gradually realized that we should re-examine the importance of the epididymis for sperm mat-uration and for the entire male reproductive system. Recently, epididymis related reports have tended to be more regionalized and more refined. We can understand cell distribution and gene expression from the perspective of a single cell. It has been reported that different regions of the epididymis have different functions and different cell compositions, which may be determined by gene expression. Here, we comprehensively reviewed the epididymal studies. From the ana-tomical structure of the epididymis to the division of its regions, we focused on the cell compo-sition and region-specific genes differentially expressed in the epididymis. The gene expression of the epididymis extends to a series of biological processes of sperm in the epididymal cavity, and the serious effects of epididymal diseases on reproduction are discussed. Therefore, the importance of the epididymis in male reproduction is emphasized. With the maturity of single-cell spatial transcriptomics, it is hoped that the technology can be applied to epididymal research and bring about the spatiotemporal landscape of the epididymis at the single-cell level.

Autoimmune disorders and some types of blood cancer originate when B lymphocytes malfunction. In particular, when B cells produce antibodies recognizing the body’s proteins, it leads to various autoimmune disorders. And when B cells of various developmental stages transform into cancer cells, it results in blood cancers, including multiple myeloma, lymphoma, and leukemia. Thus, new methods of targeting B cells are required for various patient groups. Here, we used protein kinase inhibitors alectinib, brigatinib, ceritinib, crizotinib, entrectinib, and lorlatinib previously approved as drugs treating anaplastic lymphoma kinase (ALK)-positive lung cancer cells. We hypothesized that the same inhibitors will efficiently target leukocyte tyrosine kinase (LTK)-positive, actively protein-secreting mature B lymphocytes, including plasma cells. We isolated CD19-positive human B cells from the blood of healthy donors and used two alternative methods to stimulate cell maturation toward plasma cells. Using cell proliferation and flow cytometry assays, we found that ceritinib and entrectinib eliminate plasma cells from B cell populations. Alectinib, brigatinib, and crizotinib also inhibited B cell proliferation, while lorlatinib had no or limited effect on B cells. More generally, we concluded that several drugs previously developed to treat ALK-positive malignant cells can be also used to treat LTK-positive B cells.

Ovine footrot is a complex multifactorial infectious disease, causing lameness in sheep with major welfare and economic consequences. Dichelobacter nodosus is the main causative bacterium, however, footrot is a polymicrobial disease with Fusobacterium necrophorum, Mycoplasma fermentans and Porphyromonas asaccharolytica also associated. There is limited understanding of the host response involved. Proinflammatory mediators interleukin (IL)-1β and C-X-C Motif Chemokine Ligand 8 (CXCL8) have been shown to play a role in the early response to D. nodosus in dermal fibroblasts and interdigital skin explant models. To further understand the response of ovine skin to bacterial stimulation, and to build the understanding of the role of the cytokines and chemokines identified in transcriptomic data, primary ovine interdigital fibroblasts and keratinocytes were isolated, cultured and stimulated with lipopolysaccharide (LPS), D. nodosus or F. necrophorum in the presence and absence of M. fermentans, whilst measuring mRNA expression and protein release of CXCL8 and, IL-1β. Stimulation with LPS, D. nodosus or F. necrophorum resulted in increased transcript levels of IL-1β and CXCL8 in M. fermentans free cells, however, only an increase in CXCL8 protein release was observed. No IL-1β protein release was detected despite increases in IL-1β mRNA, suggesting the signal for intracellular pre-IL-1β processing may be lacking when culturing primary cells in isolation. Keratinocytes and fibroblasts naturally infected with M. fermentans showed little response to LPS, a range of D. nodosus preparations or heat-inactivated F. necrophorum. Primary single cell culture models complement ex vivo organ culture models to study different aspects of the host response to D. nodosus. Ovine keratinocytes and fibroblasts infected with M. fermentans had a reduced response to experimental bacterial stimulation. However, in the case of footrot where Mycoplasma spp. are associated with diseased feet, this natural infection gives important insights into the impact of multiple pathogens on the host response.

Thermodynamic studies consider living entities as dissipative structures. Organisms maintain and develop an orderly structure by exchanging matter, energy, and entropy with the surrounding environment; thus, maintaining life and growth. For a single cell, the temperature, volume, content concentration, and content complexity are four control variables. For warm-blooded animals, both temperature and content concentration are assumed to be constant, and only volume and content complexity, i.e., various proteins, nucleic acids, and small molecular substances in the cell and their interactions, are considered as acting variables. Thus, the potential function of a single cell should conform to the cusp catastrophe model. As the studies on the specific mathematical models of the relationship between the volume and content complexity are not available, we could not propose specific methods for the specific variants of the potential function of this cusp catastrophe model. We could only present our approximate results based on the basic characteristics of the cusp catastrophe model. We speculated that when a single cell is in a stable state, it cannot undergo differentiation, dedifferentiation, and division. These behaviors occur only when the cell enters an unstable state. Based on this speculation, we divided somatic cells of warm-blooded animals into two types, namely stable cells and non-steady cells. If we consider a warm-blooded animal as a whole dissipative structure, its control variables should have steady-state cells, non-steady-state cells, and negative entropy input. If we assume that the negative entropy input is constant, the proportion of non-steady cells and the total number of cells can be used as the control variables of the potential function. For warm-blooded animals, their potential function also conforms to the cusp catastrophe model. Because the studies on the relationship between the proportion of non-steady-state cells and the total cell number are rare, we could not propose specific methods for the variation of the potential function of this cusp catastrophe model. We could only present our approximate results according to the basic characteristics of the cusp catastrophe model. We speculated that as individuals, animals should be in a stable state during development. Once they enter an unstable state, they will fall ill or die. For humans, the proportion of non-steady cells decreases during the growth process from a fertilized egg to old age. From the fertilized egg to adulthood, the total cell number increases; however, in old age, the total cell number begins to decrease gradually. The entire developmental curve will gradually enter an unstable state. We speculated that once the developmental curve of a human enters an unstable state, it is death for the elderly.

Cell death by apoptosis is a major cellular response, in the control of tissue homeostasis and as a defense mechanism in case of cellular aggression like an infection. Cell self-destruction is part of antiviral responses, aimed at limiting the spread of a virus. Although it may contribute to the deleterious effects in infectious pathology, apoptosis remains a key mechanism for viral clearance and resolution of infection. The control mechanisms of cell death processes by viruses have been extensively studied. Apoptosis can be triggered by different viral determinants, through different pathways, as a result of virally induced cell stresses and innate immune responses. Zika virus (ZIKV) induces Zika disease in humans which has caused severe neurological forms, birth defects and microcephaly in newborns during the last epidemics. ZIKV also surprised by revealing an ability to persist in the genital tract and in semen, thus being sexually transmitted. Mechanisms of diverting antiviral responses such as the interferon response, the role of cytopathic effects and apoptosis in the etiology of the disease have been widely studied and debated. In this review, we examined the interplay between ZIKV infection of different cell types and apoptosis and how the virus deals with this cellular response. We illustrate a duality in the effects of ZIKV-controlled apoptosis, depending on whether it occurs too early or too late, respectively in neuropathogenesis, or in long-term viral persistence. We further discuss a prospective role for apoptosis in ZIKV-related therapies, and the use of ZIKV as an oncolytic agent.

Light microscopy is indispensable for analysis of bacterial spatial organization. However, imaging in bacteria is difficult due to their small sizes and the fact that most species are non-spherical, meaning they typically lie horizontally on a microscope coverslip. This is especially problematic when considering that many essential bacterial processes—such as cell division—occur along the short axes of these cells, and so are viewed side-on by standard microscopy. We recently developed a pair of methods to overcome this problem by forcing cells to stand vertically during imaging, named VerCINI (Vertical Cell Imaging by Nanostructured Immobilisation) and µVerCINI (Microfluidic VerCINI). The concept behind both methods is that cells are imaged while confined vertically inside cell traps made from a nanofabricated mould. By doing so, the short axes of the cells are rotated parallel to the microscope imaging plane and are imaged with high resolution. μVerCINI combines the vertical cell confinement with microfluidics so that vertical imaging can be done during fluid exchange, such as during antibiotic perturbations. Here, we provide a practical guide to implementing both VerCINI and µVerCINI, with detailed protocols and experience-based tips so that interested researchers can easily use one or both imaging methods to complement their current approaches.

Background: Thousands of genetic variants have been associated with hematological traits, though target genes remain unknown at most loci. Also, limited analyses have been conducted in African ancestry and Hispanic/Latino populations; hematological trait associated variants more common in these populations have likely been missed. Methods: To derive gene expression prediction models, we used ancestry-stratified datasets from the Multi-Ethnic Study of Atherosclerosis (MESA, including N=229 African American and N=381 Hispanic/Latino participants, monocytes) and the Depression Genes and Networks study (DGN, N = 922 European ancestry participants, whole blood). We then performed a transcriptome-wide association study (TWAS) for platelet count, hemoglobin, hematocrit, and white blood cell count in African (N = 27,955) and Hispanic/Latino (N = 28,324) ancestry participants. Results: Our results revealed 24 suggestive signals (p < 1×10^(-4)) that were conditionally distinct from known GWAS identified variants and successfully replicated these signals in European ancestry subjects from UK Biobank. We found modestly improved correlation of predicted and measured gene expression in an independent African American cohort (the Genetic Epidemiology Network of Arteriopathy (GENOA) study (N=802), lymphoblastoid cell lines) using the larger DGN reference panel; however, some genes were well predicted using MESA but not DGN. Conclusions: These analyses demonstrate the importance of performing TWAS and other genetic analyses across diverse populations and of balancing sample size and ancestry background matching when selecting a TWAS reference panel.

Unanticipated errors in scientific research data can be attributed to the unwarranted assumption of uniformity in the polystyrene surface that is ubiquitously used in tissue culture flasks and dishes. We have shown that when adherent cells are subjected to fluid shear force, equivalent to rinsing the culture with a balanced salt solution, cells on some areas of the polystyrene surface will immediately rupture while still adherent on the surface. This heterogeneity on the polystyrene surface can cause unexpected variability in experimental results and in replicating experiments among labs. In this paper a novel quantitative method is described to measure the degree of heterogeneity on the polystyrene surface of tissue culture flasks. The results show significant variation among several brands of tissue culture flasks as well as large variability within the production lot of a manufacturer. The assay method involves loading the cells with a fluorescent marker that is released upon membrane rupture. Cell membrane rupture also causes the loss of marker proteins used in Westernblots. This novel assay method can be used to monitor the batch consistency and the manufacturing process of flasks and dishes. It may also be used to test new biomaterials.

In this study, the elastic properties of composite materials are investigated, considering the effects of separation of fiber-matrix joint boundary and matrix failure. In this method, by assuming periodic microstructure and using a linear approximation of the displacement field by applying continuity and equilibrium conditions, the composite fiber composite relation is determined. The effect of separation is assumed by introducing tangential and normal scalar parameters in the equations by assuming the displacement field jump at the common boundary. In order to express the effect of matrix micro-cracks, the fracture mechanics framework of continuous environments was used and the micro-cracks parallel to the fibers, perpendicular to the fibers and in the thickness direction with scalar parameters were expressed. At the end of the effect of these parameters the results are presented in graphs. The results show that the presence of defective joint at the joint boundary and the matrix micro-components reduce the hardness of the composite and thus it’s bearing load, which is more significant at the defective joint state.

The paper presents the author’ original modification of the residual method in order to construct an efficient tool for assessing the condition of photovoltaic cells, in particular – panels made of amorphous cells which are characterized by significant random operation parameter changes. The solution constructed has been verified through experiments, which show the usefulness of the proposed methodology as it allows to minimize the risk of obtaining false diagnostic information in the process of assessing the condition of photovoltaic panels.

Excretion and osmoregulation are fundamental processes of the organism, as they prevent the accumulation of toxic waste products in the body and control the osmotic differences between the cells and the environment. In most of the animals these phenomena are taking place through specialized organs, namely excretory organs, composed of diverse cell types that are performing tasks such as secretion and ultrafiltration. Although the morphology and embryology of excretory organs can differ dramatically, the common spatial arrangement of structural proteins and transporters as well as the similar transcriptional developmental programs underlying their formation suggests the homology of their cell types. In this chapter we discuss the current understanding of the evolution of excretory organsfrom a comparative morphological, developmental and functional perspective, flanked by an additional, cell-type perspective. We argue that a putative homologization of certain excretory cell types does not necessarily reflect the homology of the resulting organs, and that integrating all different levels of comparison is crucial for addressing evolutionary questions.

Multinucleate cells can be produced in Dictyostelium by electric-pulse induced fusion. In these cells unilateral cleavage furrows are formed at spaces between areas that are controlled by aster microtubules. A peculiarity of unilateral cleavage furrows is their propensity to join laterally with other furrows into rings to form constrictions. This means, cytokinesis is biphasic in multinucleate cells, the final abscission of daughter cells being independent of the initial direction of furrow progression. Myosin-II and the actin-filament cross-linking protein cortexillin accumulate in the unilateral furrows, as they do in the normal cleavage furrows of mononucleate cells. Myosin-II is not essential for cytokinesis, but stabilizes and confines the position of the cleavage furrows.

Bio-devices are designed to allow biological matter to perform in vitro almost the same functions it performs in vivo. Therefore, they can benefit from the specificities of such materials and are expected to perform better than traditional devices. On the other hand, the integration of biological material with electronic/electrochemical instrumentation requires careful attention and can produce unconventional results. In this paper, we describe the impedance response of an electrochemical cell that converts sunlight into electrical power. It uses the photosynthetic system known as Reaction Center, which is the core of photosynthesis in several living beings. Under illumination, an abrupt transformation drives the cell electrical response from insulator to conductor and a photocurrent is observed. The impedance spectrum shows a peculiar shape which significantly modifies with the protein activation. It has been analyzed by means of a graphical/analytical/ numerical procedure. The modelling identifies an analogue electrical circuit, whose parameters give quantitative information about the underlying process. Finally, an appropriate normalization of data is proposed which validates data in dark and light and can be useful as a fast screening of measurements.

Recently, some authors have developed a shell-less culture system for chick embryos by using a plastic lm as culture vessels. We will show that velocity of rotation of vessels in incubator and temperature have direct relations with growth of chick embryos. We also obtain a mathematical relation between velocity, temperature and rate of growth of chick embryo.

The wide range of applications of nanoparticles (NPs) has increased the probability of environmental and occupational exposure. In a previous comparative study on cell line A549, we observed that oxidative stress caused by Co₃O₄ NPs affects the energetic homeostasis and the detoxification capacity, preventing autophagy induced by TiO₂ NPs. In this study, we have investigated the effects of NPs on the cell cycle.Cytofluorimetric analysis showed a slow-down of the cell cycle progression for both NPs, with increases in the percentage of resting cells in the G0/G1. These observations were confirmed by a reduced expression of all cyclins, especially of CCNE1 and CDK2, involved in the late stages of the G1 phase, coupled with a significant increase in the expression of p21 only for Co₃O₄ NP exposure. On the contrary, the effects of TiO₂ NPs were modest. Cell cycle related miRNA-34a, miRNA-126 and miRNA-1290 resulted increased at different early time-points (4-8h) but were down-expressed at 24h-48h only after TiO₂ NP exposure. Our results show that NPs have an antiproliferative effect and deregulate cell cycle on A549 cells. These effects should not be underestimated because regulation of cycle progression is crucial for cell survival and repair of genetic damage.

The development of plant-based meat analogues has become a significant challenge for the food industry in recent years due to the increasing demand for sustainable and healthier proteins in the context of a global protein transition. Plant-based meat analogues imitate the visual, textural, and chemical properties of traditional meat products and are required to closely resemble meat to appeal to consumers. In addition, consumers demand natural, clean-label, and nutritional and healthy products. To address these challenges, the food industry must develop highly healthy, nutritious, and E-number-free meat analogue products. Understanding the functionality of each ingredient and its role in the food matrix is crucial to being a key player in the innovation of the meat analogues market. This review provides updated information on the primary ingredients utilized for the development of plant-based burger meat alternatives and their functionality. The key components of meat analogue burgers are outlined, including plant proteins, binding agents, fats and oils, flavorings, colorings, preservatives, fortificants, and clean-label considerations.

This paper presents the PEnsemble 4 model, a sophisticated machine learning framework that integrates IoT-based environmental data to accurately forecast maize yield. With the projected significant growth in global maize demand over the next decade, the inherent risks posed by the crop’s dependence on weather conditions necessitate improved prediction capabilities. The PEnsemble 4 model, developed with high accuracy, incorporates comprehensive datasets encompassing soil attributes, nutrient composition, weather conditions, and UAV-captured vegetation imagery. By employing a combination of Huber and M estimates, the PEnsemble 4 model effectively analyzes temporal patterns in vegetation indices, specifically CIre and NDRE, which serve as reliable indicators of canopy density and plant height. In addition, this research significantly contributes to precision agriculture by offering an efficient and sustainable alternative to conventional farming practices through precise yield predictions. Notably, the PEnsemble 4 model enables earlier estimation, advancing the timeline for yield prediction from the conventional day 100 in the R6 stage to day 79 in the R2 stage. This improvement enhances decision-making processes in farming operations. The remarkable accuracy rate of 91% underscores the importance of adopting a multifaceted data approach that harnesses IoT-derived environmental insights. Additionally, the PEnsemble 4 model extends its benefits beyond yield prediction, facilitating the detection of water and crop stress, as well as disease monitoring in broader agricultural contexts. Ultimately, the PEnsemble 4 model establishes a new standard in maize yield prediction, revolutionizing crop management and protection through the synergistic utilization of IoT and machine learning technologies.

Many complex behaviours in biological systems emerge from large populations of interacting molecules or cells, generating functions that go beyond the capabilities of the individual parts. Such collective phenomena are of great interest to bioengineers due to their robustness and scalability. However, engineering emergent collective functions is difficult because they arise as a consequence of complex multi-level feedback, which often spans multiple length-scales. Here, we present a perspective on how some of these challenges could be overcome by using multi-agent modelling as a design framework within synthetic biology. Using case studies covering the construction of synthetic ecologies to biological computation and synthetic cellularity, we show how multi-agent modelling can capture the core features of complex multi-scale systems and provide novel insights into the underlying mechanisms which guide emergent functionalities across scales. The ability to unravel design rules underpinning these behaviours offers a means to take synthetic biology beyond single molecules or cells and towards the creation of systems with functions that can only emerge from collectives at multiple scales.

Adult mesenchymal stem cells (MSCs) are a promising cell source for tissue regeneration. However, ex vivo expansion results in cell senescence; cells lose their proliferation and differentiation capacity. Fetal MSCs can be an alternative due to their robust proliferation and differentiation capacities as well as immune privilege properties. Given the rejuvenation effect of decellularized extracellular matrix (dECM) on adult MSCs, it remains unknown whether dECM influences the regenerative potential of fetal stem cells. In this study, passage five fetal nucleus pulposus cells (fNPCs) and fetal synovium-derived stem cells (fSDSCs) were expanded on dECMs deposited by fNPCs (NECM) and fSDSCs (SECM) for one passage with expansion on tissue culture plastic (Plastic) as a control. We found that dECM expanded fNPCs and fSDSCs exhibited both similarities and differences in expression of stemness genes and surface markers. Expanded fNPCs yielded more differentiated pellets after chondrogenic induction but no adipogenic differentiation following adipogenic induction in both Plastic and dECM groups than the corresponding fSDSC group. Despite a significant increase in fNPCs, dECM expanded fSDSCs exhibited no increase in chondrogenic potential; however, compared to the Plastic group, dECM expanded fSDSCs exhibited a small increase in osteogenic potential and a great increase in adipogenic potential. These results suggest that fNPCs are more sensitive to NECM rejuvenation for cartilage tissue engineering and regeneration; in contrast, dECMs exhibited limited effects on fSDSC rejuvenation in chondrogenic capacity except for enhanced adipogenic capacity following expansion on SECM.

Adult stem cells (ASCs) can be cultured with difficulty from most tissues, often requiring chemical or transgenic modification to achieve adequate quantities. We show here that mouse primary fibroblasts grown in suspension change from the elongated and flattened morphology observed under standard adherent culture conditions generating rounded cells with large nuclei and scant cytoplasm expressing the mesenchymal stem cell (MSC) marker (Sca1; Ly6A) within 24hrs. Based on this initial observation, we describe here a suspension culture method that, irrespective of the lineage used, mouse fibroblast, primary human somatic cells (fibroblasts, hepatocytes and keratinocytes), is capable of generating a high yield of cells in spheroid form which display expression of ASCs surface markers, circumventing the anoikis which often occurs at this stage. Moreover, mouse fibroblasts-derived spheroids can be differentiated into adipogenic and osteogenic lineages. Analysis of single cell RNA sequence data identified 8 distinct cell clusters with one in particular comprising approximately 10% of the cells showing high levels of proliferative capacity expressing high levels of genes related to MSCs and self-renewal as well as extracellular matrix (ECM). We believe the rapid, high-yield generation of proliferative, multi-potent ASC-like cells by the process we term suspension-induced stem cell transition (SIST) could have significant implications for regenerative medicine.

An automatic recognizing system of white blood cells can assist hematologists in the diagnosis of many diseases, where accuracy and efficiency are paramount for computer-based system. In this paper, we present a new image processing system to recognize the five types of white blood cells in peripheral blood with marked improvement in efficiency when juxtaposed against mainstream methods. The prevailing deep learning segmentation solutions often utilize millions of parameters to extract high-level image features and neglect the incorporation of prior domain knowledge, which consequently consume substantial computational resources and increase the risk of overfitting, especially when limited medical image samples are available for training. To address these challenges, we propose a novel memory-efficient strategy that exploits graph structures derived from the images. Specifically, we introduce a lightweight superpixel-based Graph Neural Network (GNN) and break new ground by introducing superpixel metric learning to segment nucles and cytoplasm. Remarkably, our proposed segmentation model (SMGNN) achieves state-of-the-art segmentation performance while utilizing at most 10000$\times$ less than the parameters compared to existing approaches. The subsequent segmentation-based cell type classification processes show satisfactory results that such automatic recognizing algorithms are accurate and efficient to execeute in hematological laboratories.

Background: Although metastatic GCT is highly curable with initial cisplatin-based chemotherapy (CT), 20-30% of patients relapse. Salvage CT options include conventional (CDCT) and high dose chemotherapy (HDCT), however definitive comparative data remains lacking. We aimed to characterize the contemporary practice patterns of salvage CT across Canada. Methods: We conducted a 30-question online survey for Canadian medical and hematological oncologists with experience in treating GCT, assessing treatment availability, patient selection, and management strategies used for relapsed GCT patients. Results: Respondents included 30 staff; from 18 cancer centers across provinces. The most common CDCT regimens used were TIP (64%) and VIP (25%). HDCT was available for (70%) in 13 centers. HDCT regimen used included carboplatin and etoposide for 2 cycles (76% in 7 centers), 3 cycles (6% in two centers), and the TICE protocol (11%, in 2 centers). “Bridging” CDCT was used by 65% respondents. Post HDCT treatments considered include surgical resection for residual disease (87.5%), maintenance etoposide (6.3%) and sur-veillance only (6.3%). Conclusions: HDCT is the most commonly used GCT salvage strategy in Canada. Significant differences exist in the treatment availability, selection and delivery of HDCT, highlighting the need for standardization of care for patients with relapsed testicular GCT.

Calcium-activated potassium channels (KCa) are the important participants in calcium signaling pathways due to their ability to be activated by increase of the intracellular free calcium concentration. KCa channels are involved in the regulation of various processes in the cells under normal, as well as pathophysiological conditions, including oncotransformation. Previously, with the use of patch-clamp method, we registered the KCa activity in the plasma membrane of human myeloid leukemia K562 cell line. Here, we performed the molecular and functional identification of KCa channels and have uncovered their role in proliferation, migration and invasion of K562 cells. Using a combined approach, we identified the functional activity of SK2, SK3 and IK channels in the plasma membrane of the cells. Selective SK and IK channel inhibitors, apamin and TRAM-34, reduced the proliferative, migratory and invasive capabilities of human myeloid leukemia cells. At the same time, the viability of K562 cells was not affected by KCa channel inhibitors. Our data imply that SK/IK channel inhibitors could be used to slow down the proliferation and spreading of leukemia cells that express functionally active KCa channels in the plasma membrane.

The most widely spread and neglected zoonotic disease – Leptospirosis, with an estimated 1.03 million cases and 58,900 deaths occurring annually, is a concern and can be prevented only by effective immunization. Due to the urgent need for a novel vaccine, various research organizations have been evaluating the protective immune response elicited by recombinant vaccines. In this study amino-acid sequences of Outermembrane protein (OMPL) 1, LipL 32, 41 and 46 from ten pathogenic serovars of Leptospira were selected to obtain epitopes that induce CD4+ and CD8+ T cell responses by binding to the MHC molecules. These B cell and T cell epitopes were used to design multi-epitope vaccine to which an adjuvant sequence was added at the N-terminal end and appropriate linkers were used to join the different epitopes to increase the efficiency of the vaccine constructs. Using an immuno-informatics approach, constructs were analyzed for the physiochemical properties, secondary and tertiary structure and its validation, docking with different receptors and prediction of binding affinity for each docked complex, followed by molecular dynamics simulation. Taking in consideration all the results, multi-epitope constructs designed may prove to be promising vaccine candidates against Leptospirosis and warrants experimental validation.

Epithelial-mesenchymal transition (EMT) networks are essential in acquiring the drug resistance and cancer malignant features in cancer stem cells (CSCs). In this regard, gene expression profiles in diffuse- and intestinal-type gastric cancer (GC) have been analyzed to reveal the network pathways in EMT and CSCs, since the diffuse-type GC has much more mesenchymal features than intestinal-type GC that has the intestinal features. The study results revealed that the activation state of several canonical pathways related to cell cycle regulation was altered. The canonical pathway on Cell cycle: G1/S checkpoint regulation was activated in diffuse-type GC, and canonical pathways on Cell cycle control of chromosomal replication and Cyclins and cell cycle regulation were activated in intestinal-type GC. Canonical pathway related to Role of BRCA1 in DNA damage response was activated in intestinal-type GC, where BRCA1, which is related to G1/S phase transition was up-regulated in intestinal-type GC. Several microRNAs (miRNAs), including mir-10, mir-17, mir-19, mir-194, mir-224, mir-25, mir-34, mir-451, and mir-605, were identified to have direct relationships of RNA-RNA interaction in Cell cycle: G1/S checkpoint regulation pathway. Additionally, cell cycle regulation may be altered in EMT conditions. The alterations in activation states of the pathways related to cell cycle regulation in diffuse- and intestinal-type GC would indicate the significance of cell cycle regulation in EMT.

Progression of oral squamous cell carcinoma (OSCC) has been associated with an escape of tumor cells from the host immune surveillance due to an increased knowledge of its underlying molecular mechanisms and its modulation by the tumor microenvironment and immune cell repertoire. In this study the expression of HLA class I (HLA-I) antigens and of components of the antigen processing machinery (APM) was analyzed in 160 pathologically classified human papilloma virus (HPV)-negative OSCC lesions and correlated to the intra-tumoral immune cell response, IFN- signaling and to the patients outcome. A heterogeneous, but predominantly lower constitutive protein expression of HLA-I APM components was found in OSCC sections when compared to non-neoplastic cells. Tumoral HLA-I APM component expression was further categorized into the three major phenotypes HLA-Ihigh/APMhigh, HLA-Ilow/APMlow and HLA-Idiscordant high/low/APMhigh. In the HLA-Ihigh/APMhigh group, the highest frequency of intra-tumoral CD8+ T cells and lowest number of CD8+ T cells close to FoxP3+ cells was found. Patients within this group presented the most unfavorable survival, which was significantly evident in stage T2 tumors. Despite a correlation with the number of intra-tumoral CD8+ T cells, tumoral JAK1 expression as a surrogate marker for IFN- signaling was not associated with HLA-I/APM expression. Thus, the presented findings strongly indicate the presence of additional factors involved in the immunomodulatory process of HPV-negative OSCC with a possible tumor-burden-dependent complex network of immune escape mechanisms beyond HLA-I/APM components and T cell infiltration in this tumor entity.

Progression of oral squamous cell carcinoma (OSCC) has been associated with an escape of tumor cells from the host immune surveillance with growing evidence of its underlying molecular mechanisms and its interaction with the immune cell control. In this study the expression of HLA class I (HLA-I) antigens and of components of the antigen processing machinery (APM) was analyzed in 160 consecutive human papilloma virus (HPV)-negative OSCC lesions and correlated to tumor specific parameters, the intratumoral immune cell response and to the patients outcome. A heterogeneous, but predominantly lower constitutive protein expression of HLA-I APM components was seen in OSCC sections when compared to non-neoplastic cells. Based on the expression levels of HLA-I APM components three main OSCC subgroups were detected and categorized into HLA-Ihigh/APMhigh, HLA-Ilow/APMlow and HLA-Idiscordant high/low/APMhigh phenotypes. In the HLA-Ihigh/APMhigh group, the highest frequency of intratumoral CD8+ T cells and lowest number of CD8+ T cells close to FoxP3 cells was found. Despite being associated with the highest T cell infiltration, patients within this group presented the most unfavorable survival, which was most evident in stage T2 tumors. Thus, the presented findings strongly indicate the presence of additional factors involved in the immunomodulatory process of HPV-negative OSCC with a possible tumor-burden-dependent complex network of immune escape mechanisms beyond HLA-I/APM components and T cell infiltration in this tumor entity.

Coronavirus disease (COVID-19) is a new discovered strain where WHO officially declares the disease as COVID-19 while the virus responsible for it called Severe Acute Respiratory Syndrome Coronavirus 2 or SARS-CoV-2. The incubation period of this disease is between 14 days. Ordinary clinical symptoms that reported around the world include fever, cough, fatigue, diarrhoea and vomiting as well as asymptomatic for certain people. Infection is spread mainly through broad droplets. In early March 2020, WHO again has announced that COVID-19 is a pandemic with currently no specific treatment. The potential use of SARS-COV-2 proteome as a vaccine candidate by analysing through B-cell and T-cell antigenicity by using a immunoinformatics approach as a vaccine development early stage. In this study, we used consensus sequence for SARS-COV-2 proteome that was retrieved from NCBI database. VaxiJen 2.0 was mainly used to identify the antigenic property of SARS-COV-2 proteins. IEDB then used to analyse the B-cell epitope, the presence of T cell immunogenic epitope in SARS-COV-2 proteins was obtained by using compromise method of MHC class I and II tools that accessible respectively using ProPred-1 server and MHC II Binding Prediction in IEDB database. The best epitopes of B and T-cell epitopes were predicted with high antigencity and the information is disseminated through web-based database resource (https://covid-19.omicstutorials.com/epitopes/). This study will be useful to find a new epitope-based candidate for SARS-COV-2. However, further study needs to be done for the next stages of vaccine development.

Malignant tumors behave dynamically as cell communities governed by ecological principles. Massive sequencing tools are unveiling the true dimension of the heterogeneity of these communities along their evolution in most human neoplasms, clear cell renal cell carcinomas (CCRCC) included. Although initially thought to be purely stochastic processes, very recent genomic analyses have shown that temporal tumor evolution in CCRCC may follow some deterministic pathways that give rise to different clones and sub-clones randomly spatially distributed across the tumor. This fact makes each case unique, unrepeatable and unpredictable. Precise and complete molecular information is crucial for patients with cancer since it may help in establishing a personalized therapy. Intratumor heterogeneity (ITH) detection relies on the correctness of tumor sampling and this is part of the pathologist’s daily work. International protocols for tumor sampling are insufficient today. They were conceived decades ago, when ITH was not an issue, and have remained unchanged until now. Noteworthy, an alternative and more efficient sampling method for detecting ITH has been developed recently. This new method, called multisite tumor sampling (MSTS), is specifically addressed to large tumors that are impossible to be totally sampled, and represent an opportunity to improve ITH detection without extra costs.

In the last two decades, the discovery of various pathways involved in renal cell carcinoma (RCC) have led to the development of biologically-driven targeted therapies. Hypoxia inducible factors (HIFs), angiogenic growth factors, von Hippel-Lindau (VHL) gene mutations and oncogenic miRNAs play essential roles in the pathogenesis and drug resistance of clear cell renal cell carcinoma. These insights have led to the development of VEGF inhibitors, mTOR inhibitors and immunotherapeutic agents which have significantly improved outcomes of patients with advanced RCC. HIF inhibitors will be a valuable asset in the growing therapeutic armamentarium of RCC. Various histone deacetylase (HDAC)inhibitors, including selenium and agents such as PT2385 and PT2977, are being explored in various clinical trials as potential HIF inhibitors to ameliorate the outcomes of RCC patients. In this article, we will review the current treatment options and highlight the potential role of selenium in the modulation of drug resistance biomarkers expressed in ccRCC tumors.

A single Information security is of pivotal concern for consistently streaming information over the widespread internetwork. The bottleneck flow of incoming and outgoing data traffic introduces the issue of malicious activities taken place by intruders, hackers and attackers in the form of authenticity desecration, gridlocking data traffic, vandalizing data and crashing the established network. The issue of emerging suspicious activities is managed by the domain of Intrusion Detection Systems (IDS). The IDS consistently monitors the network for identifica-tion of suspicious activities and generates alarm and indication in presence of malicious threats and worms. The performance of IDS is improved by using different signature based machine learning algorithms. In this paper, the performance of IDS model is determined using hybridization of nestrov-accelerated adaptive moment estimation –stochastic gradient descent (HNADAM-SDG) algorithm. The performance of the algorithm is compared with other classi-fication algorithms as logistic regression, ridge classifier and ensemble algorithm by adapting feature selection and optimization techniques