Excess associated gas from unconventional wells is typically flared while excess produced water is injected underground. In this work, flare gas recovery is integrated with produced water desalination and a solar pre-heater. The solar module with a beam splitter preheats the produced water. Aspen Plus was used to perform process modeling as well as to estimate capital and operating costs. The Solar-Flare Gas RDcovery- desalination (Solar-FGRD) process can conserve water resources and reduce the brine injection by 77%. The accompanying solar farm results in excess solar electricity for exporting to the grid. The process burner combustion efficiency (CE) is 99.8% with the destruction and removal efficiency (DRE) of 99.99% for methane as opposed to the flare CE of 80-98% (and a methane DRE of 91-98%). The greenhouse gas (GHG) emissions for CO2 and methane in terms of CO2 equivalent (CO2e) can be reduced by 45 % for US North Dakota & Texas flaring and 13% for North Sea flaring by employing the Solar- FGRD process. The comprehensive financial analysis demonstrates the financial-economic feasibility of the investment project with or without tax credits. Best-case and worst-case scenarios provide a realistic range that investors can consider before making investment decisions.

Vascular endothelial growth factor (VEGF) and its receptors (VEGFRs) serve an essential role in tumor angiogenesis and have emerged as potential therapeutic targets in lung cancer. This review explores the significance of serum VEGF levels as a predictive biomarker in non-small cell lung cancer (NSCLC). The VEGF family, consisting of VEGFA, VEGFB, VEGFC, VEGFD, and placenta growth factor (PlGF), engages with specific receptors, including tyrosine kinase receptors (VEGFR-1, VEGFR-2, and VEGFR-3) and neuropilin receptors (NRP-1 and NRP-2), to promote angiogenesis and lymphangiogenesis. VEGF-A, the primary component of the VEGF family, binds to VEGFR-2 to stimulate endothelial cell proliferation and migration, while VEGF-B, VEGF-C, and VEGF-D interact with VEGFR-1 and VEGFR-3 to regulate tumor angiogenesis, lymphangiogenesis, and metastasis. The VEGF/VEGFR signaling pathway activates various downstream effectors, including phospholipase Cγ1, MAPK, and PI3K/Akt, which are essential for maintaining vascular homeostasis and promoting angiogenesis. In NSCLC, elevated serum VEGF levels have been observed, and the VEGF/VEGFR axis is frequently impaired, leading to irregular blood vessel formation and metastatic spread. Despite the development of anti-VEGF therapies, their impact on lung cancer outcomes has been limited. Further research is needed to optimize the effectiveness of these treatments and elucidate the potential of serum VEGF as a predictive biomarker in NSCLC.

Transitional cell carcinoma of the bladder is currently a very representative model of development in basic, pathogenic and therapeutic research within the framework of modern clinical oncology. The article aims to present a clinical case of transitional cell carcinoma of the bladder, a diagnostic approach that takes into consideration the combination of clinical imaging methods, for which a 67-year-old white male patient with a history of macroscopic hematuria is presented, which is why which is treated and placed under follow-up, demonstrating that a successful diagnostic approach has been, is and will always be a fascinating intellectual challenge for doctors, and an inexhaustible source of satisfaction in their professional life, as long as its bases and procedures are mastered. . , this serves as the uniqueness and novelty of the work that is presented.

Cutaneous Squamous Cell Carcinoma (cSCC) is an increasingly common malignancy of the skin and the leading cause of death from skin cancer in adults over the age of 85. Fibroblast Growth Factor Receptor 2 (FGFR2) has been identified as an important effector of signaling pathways that lead to the growth and development of cSCC. In recent years, there have been numerous studies evaluating the role FGFR2 plays in multiple cancers, its contribution to resistance to anti-cancer therapy, and new drugs that may be used to inhibit FGFR2. This review will provide an overview of our current understanding of FGFR2 and potential mechanisms in which we can target FGFR2 in cSCC. The goals of this review are the following: 1) to highlight our current knowledge on the role of FGFR2 in healthy skin and contrast this with its role in the development of cancer; 2) to further explain the specific molecular mechanisms that FGFR2 uses to promote tumorigenesis; 3) to describe how FGFR2 contributes to more invasive disease, 4) to describe its immunosuppressive effects in skin, and 5) to evaluate its effect on current anticancer therapy and discuss therapies on the horizon to target FGFR2 related malignancy.

Melanoma, a deadly form of skin cancer, has seen improved survival rates due to advances in diagnosis and treatment, yet the need for further improvement remains critical. Tumor-associated antigens, such as PRAME (Preferentially Expressed Antigen in Melanoma), offer promising avenues for enhanced diagnostic precision, prognostic assessment, and targeted immunotherapy. PRAME, a cancer-testis antigen, is selectively expressed in various cancers, including melanoma, and plays a key role in promoting tumorigenesis through inhibition of retinoic acid signaling, epithelial-to-mesenchymal transition, and immune evasion. This review explores the diagnostic utility of PRAME in distinguishing melanoma from benign nevi, its prognostic value in aggressive melanoma subtypes, and its potential as a therapeutic target in cancer vaccines and adoptive T-cell therapies. While PRAME-targeted therapies face challenges such as tumor heterogeneity and immune suppression, ongoing research aims to overcome these barriers, offering hope for more effective melanoma treatments.

The growing use of proton-exchange membrane fuel cells (PEMFC) in hybrid propulsion systems, aimed at replacing traditional internal combustion engines and reducing greenhouse gas emis-sions. Effective power distribution between the fuel cell and the energy storage system (ESS) is crucial, which has led to a growing emphasis on developing energy management systems (EMS) to efficiently implement this integration. To address this goal, this study examines the performance of a fuzzy logic rule-based strategy for a hybrid fuel cell propulsion system in a small hydro-gen-powered passenger vessel. The primary objective is to optimize fuel efficiency, with particular attention to reducing hydrogen consumption. The analysis is carried out under typical operating conditions encountered during a river trip. Comparisons between the proposed strategy with other approaches —control-based, optimization-based and deterministic rule-based— are con-ducted to verify the effectiveness of the proposed strategy. Simulation results indicated that EMS based on fuzzy logic mechanisms was the most successful in reducing fuel consumption. The superior performance of this method stems from its ability to adaptively manage power distri-bution between the fuel cell and energy storage systems.

It is generally assumed that spirochetemia does not take place at the early localized stage of Lyme disease. This misassumption is due to the lack of sensitive and specific methods for detection. This paper reports that separation of the platelet-rich plasma immediately after venous blood collection to prevent the spirochetes from invading the lymphocytes in the test tube is crucial in molecular detection of a single Borrelia burgdorferi cell spun down in platelet pellets. In 2023, we tested the venous blood specimens of 145 people residing in Lyme disease-endemic areas in the United States. In 98 of them, who were suspected of having early localized Lyme disease irrespective of the absence or presence of a skin lesion, we found 33 specimens (33.7%) positive for B. burgdorferi, including 17 positive for FlaB gene only, 15 positive for both FlaB gene and 16S rRNA gene, and 1 positive for 16S rRNA gene only. Eight (17.0%) of the 47 asymptomatic resident controls were positive for FlaB PCR only. FlaB is a more sensitive chromosomal target than the 16S rRNA gene for the detection of 1-3 B. burgdorferi cells in 1 mL of platelet-rich plasma due to spirochetes gaining or retaining FlaB paralogs at the early localized stage of Lyme disease.

Recent increases in the demand for automotive lithium-ion batteries (LIBs) have led to higher needs for critical materials like lithium, cobalt, nickel, and graphite. Consequently, recovering materials from spent batteries has gained importance. This study aimed to (1) develop a mechanical pre-treatment method for separating and concentrating materials from spent NCM622 cells, and (2) evaluate the economic, environmental, and scarcity potentials of these fractions. The pre-treatment involved grinding and granulometric separation, producing six particle-size fractions. Analysis showed that the finest fraction (n < 0.5 mm) had 85% of lithium, 77.4% of cobalt, 75.1% of manganese, and 68.5% of nickel. This fraction represented 40.4% of the processed mass and had the highest economic value (USD 3,669/ton NCM cell). This fraction was also prioritized for recycling due to its superior environmental and scarcity considerations.

Turgor pressure – the intracellular hydrostatic pressure exerted by the cell protoplast on the cell wall – is of critical physiological importance for all organisms that have the cell wall. However, in mycology there is increasing evidence that turgor pressure is not the only and not always the main factor influencing, for example, the tension of the plasma membrane, the size and shape of non-apical fungal cells, and is not even always the determining factor of apical growth. This paper characterizes various aspects of the curtain model, previously proposed to describe the regulation of plasma membrane tension in the hyphae of basidiomycetes. The current understanding of the four main components of the curtain model are outlined: the driving actin cytoskeleton, the elastic cell wall, tight adhesion of the plasma membrane to the cell wall and macroinvaginations of the plasma membrane. All four elements, summed in the single curtain model, complement or replace the physiological functions of turgor, and allow imagining how a non-apical fungal cell maintains its integrity and physiological functionality under changing osmotic and other conditions. Further experimental confirmation of the curtain model is not only of fundamental importance for mycology, but also opens up new prospects in the development of drugs that control the growth and pathogenicity of filamentous fungi.

Cellular therapies exhibit immense potential in treating complex diseases with sustained response. Manufacture of cell therapies involves purification and engineering of specific cells from a donor or patient to achieve a therapeutic response upon injection. Magnetic cell sorting targeting the presence or absence of surface markers is commonly used for upfront purification. However, emerging research is showing that optimal therapeutic phenotypes are characterized not only by the presence or absence of specific antigens but also on antigen density. Unfortunately, current cell purification tools like magnetic or fluorescence activated cell sorting lack the resolution to differentiate populations based on antigen density while maintaining scalability. Utilizing a technique known as Digital Magnetic Sorting (DMS) we demonstrate proof of concept for a scalable, magnetic based approach to fractionate cell populations based on antigen density level. Targeting CD4 on human leukocytes, DMS demonstrated fractionation into CD4High T cells and CD4Low monocytes/neutrophils as quantified by flow cytometry and single cell RNA sequencing. DMS also demonstrated high throughput processing at throughputs 5-10X faster than flow cytometry. We believe DMS can be leveraged and scaled to enable antigen density-based sorting in cell therapy manufacturing, leading to the production of more potent and sustainable cellular therapies.

The objective of this study was to evaluate the antifungal activity of eugenol against the genomic DNA of Candida albicans. A quantitative experimental study was conducted with the C. albicans INCQS 40006 strain. Eugenol was tested in concentrations of 12.5%, 25%, 50%, 75% and 100%. The test substance remained in contact with the fungus for 24 hours in a BHI medium in a microbio-logical greenhouse, then the DNA extraction was performed and part of the sample obtained was washed with TE to remove the Eugenol. The fungus was placed in BHI again and placed for 24 hours in a microbiological greenhouse. At the end, the samples with 100% eugenol went through seeds in Sabouraud Dextrose for confirmatory tests. Thus, from the electrophoresis, which admits evaluating the quality of the extracted material, preserving its characteristics and integrity, it was observed in the eugenol concentration test (test 1), in which the eugenol was in direct contact with the pathogen, that in concentrations of equal to or greater than 75% of the test substance the pathogen had no proliferative power. In which the fungus's ability to recover is evaluated (test 2), eugenol was removed from the sample, noting that in 100% of the test substance the fungus did not have the ability to recover, resulting in definitive cell death.

T-cell redirecting therapies, which include chimeric antigen receptor T-cells (CAR-T) and bispecific antibodies (BSAs), have revolutionized the treatment of relapsed\refractory large B-cell lymphoma (LBCL). Expanding clinical experience with these advanced therapies shows the potential for optimization of their use with combination or consolidation strategies, which necessitates prognostic stratification of patients. While traditional clinical prognostic factors identified in the era of chemotherapy are characterized by limited value, the tumor microenvironment (TME) is becoming a new prognostic cluster. We examine how the heterogeneity of LBCL, characterized by variations in tumor parameters and differences in TME immune cell composition, immune checkpoint expression, and cytokine milieu, correlates with both positive responses and resistance to treatment. While classical parameters such as histological subtype, cell of origin, and target antigen expression lack proven prognostic value for T-cell redirecting therapies, the density and functional state of tumor-infiltrating lymphocytes, tumor-associated macrophages, and immune checkpoint molecules are shown to be critical determinants of therapeutic success, particularly in CAR-T therapy. We identify several gaps in current knowledge and suggest that insights gained from CAR-T experience could be instrumental in refining BSA applications. This report also highlights limitations in current knowledge, as TME data derive from a limited number of registrational trials with varying methodologies, complicating cross-study comparisons and often focusing on immediate response metrics rather than long-term outcomes. By dissecting the complex interactions within the TME, this review aims to identify new prognostic factors and targets, ultimately fostering more effective and tailored treatment strategies for LBCL patients.

Hepatocellular carcinoma (HCC) is a liver cancer that originates from hepatocytes. It is a primary health issue and one of the most common malignant tumors. The US FDA approved Sorafenib as the first-line anti-cancer drug for advanced HCC patients. A few patients with advanced HCC may benefit from Sorafenib treatment, but most of the patients eventually develop resistance to Sorafenib, which leads to its poor prognosis. Long-noncoding RNAs (lncRNA) have been found to play a critical role in tumorigenesis and the development of HCC, as well as various cancers. It also plays a vital role in tumor drug resistance, while the mechanisms of LncRNAs in HCC sorafenib resistance have not been characterized yet. In this review article, we summarized the molecular mechanisms, such as the involvement of transporters, receptors, cell death regulation, and other factors impacting sorafenib resistance in HCC.

The knee joint plays a vital role in human movement, bearing mechanical loads during daily and athletic activities. This review explores the knee joint's response to mechanical loading, particularly focusing on mechanotransduction—the process by which cells convert mechanical stimuli into biochemical signals. These signals trigger cellular responses crucial for maintaining joint health, promoting repair, and adapting to mechanical changes. Understanding mechanotransduction is key to developing effective rehabilitation strategies for knee injuries and managing conditions like osteoarthritis. This review synthesizes current research on the cellular and molecular pathways involved in knee joint mechanotransduction, examining how various loading conditions—such as compression, tension, and shear—affect tissues like cartilage, synovium, ligaments, and tendons. The review discusses the impact of mechanotransduction on cellular components such as chondrocytes, synoviocytes, and fibroblasts, which are essential for maintaining tissue integrity and function. It also highlights the role of signaling pathways like MAPK, NF-κB, and Wnt in mediating cellular responses to mechanical stimuli, influencing gene expression, protein synthesis, and metabolism. By understanding these mechanisms, clinicians can develop targeted therapeutic interventions, including controlled loading protocols, exercise regimens, and manual therapies. This review underscores the importance of integrating mechanobiology insights into rehabilitation practices to enhance tissue repair, restore function, and prevent further joint degeneration, ultimately improving knee joint health and patient outcomes.

Pluripotent stem cells (PSCs) hold many potential research and clinical benefits due to their ability to differentiate into nearly every cell type in the body. They are often used as model systems for early stages of ontogenesis to better understand key developmental pathways, and for drug screening. However, in order to fully realise the potential of PSCs and their translational applications, a deeper understanding of developmental pathways, especially in humans, is required. Several signalling molecules play important roles during development and are required for proper differentiation of PSCs. The concentration and timing of signal activation is important, with perturbations resulting in improper development and/or pathology. Bone morphogenetic protein (BMP) is one such key signalling molecule involved in the specification and differentiation of various cell types and tissues in the human body including tooth and otic development. In this review, we describe the role of BMP signalling and its regulation, the consequences of BMP dysregulation in disease and differentiation, and how PSCs can be used to investigate the effects of BMP modulation during development, mainly focusing on otic development. Finally, we emphasise the unique role of BMP4 in otic specification and how refined knowledge in controlling its regulation could lead to the generation of more robust and reproducible human PSC-derived otic organoids for research and translational applications.

Background. Sarcopenia is characterized by progressive loss of muscle mass and function. It is often associated with frailty, a syndrome linked to physical disability and shortened survival in various diseases, including cancer patients. Low serum alanine aminotransferase (ALT) values, representing low muscle mass (sarcopenia), is a routine blood test that was shown to be associated with increased frailty and subsequently shortened survival in several cancers. In the current study, we aimed to test the association between low ALT and shorter survival in renal cell carcinoma (RCC) patients and survivors. Methods. This was a retrospective analysis of RCC patients and survivors, both in and out-patients. We defined patients with sarcopenia as those presenting with ALT < 17 IU/L. Results. We identified records of 3,012 patients diagnosed with RCC. The final cohort included 1,830 eligible patients (mean age 65.6 ± 13.3 years, 68% were men) of whom only 179 underwent surgical treatment. Out of the eligible cohort, 811 patients (44.3% ( had ALT < 17 IU/L. The mean ALT value of patients within the low ALT group was 11.79 IU/L while the mean value in the higher ALT level group was 24.44 IU/L (p < 0.001). Patients in the lower ALT group were older (67.9 vs. 63.7 years; p < 0.001) and had lower BMI (26.6 vs. 28; p < 0.001). In addition, patients with low ALT had lower hemoglobin values (12.14 vs. 12.91 g/dL; p < 0.001), higher serum creatinine (1.49 vs. 1.14; p < 0.001) and higher platelets to lymphocytes ratio (178 vs. 156; p < 0.001). In a univariate analysis, low ALT levels were associated with a 72% increase in mortality (95% CI 1.46-2.02, P < 0.001). In a multivariate model controlled for age, male gender, hemoglobin, platelets, LDH, neutrophil to lymphocytes ratio and platelets to lymphocytes ratio, low ALT levels were associated with a 27% increase in mortality (HR = 1.27, 95% CI 1.08 – 1.51; p = 0.005]. Conclusion. Low ALT values, indicative of sarcopenia and frailty, are associated with shortened survival of RCC patients and survivors and could potentially be applied for optimizing individual treatment decisions.

Lignin is nature's second most abundant vascular plant biopolymer, playing significant roles in mechanical support, water transport, and stress responses. This study identified 90 lignin biosynthesis genes in the rice based on phylogeny and motif constitution, and they belong to PAL, C4H, 4CL, HCT, C3H, CCoAOMT, CCR, F5H, COMT, and CAD families. Duplication events contributed largely to the expansion of these gene families, such as PAL, CCoAOMT, CCR, and CAD families mainly attributed to tandem and segmental duplication, respectively. Microarray data of 33 tissue samples covering the entire life cycle of rice suggested, fairly high PAL, HCT, C3H, CCoAOMT, CCR, COMT, and CAD gene expression, and rather variable C4H, 4CL, and F5H expression. Some members of lignin-related genes (OsCCRL11, OsHCT1/2/5, OsCCoAOMT1/3/5, OsCOMT, OsC3H, OsCAD2, and OsPAL1/6) were expressed in all tissues examined. The expression patterns of lignin-related genes can be divided into two major groups with eight subgroups, each showing a distinct co-expression in tissues representing typically primary and secondary cell wall constitutions. Some lignin-related genes were strongly co-expressed in tissues typical of secondary cell walls. The combined HPLC analysis showed increased lignin monomers (H, G, and S) in five genotypes from young to old growth stages. Based on 90 genes' microarray data, 27 genes were selected for qRT-PCR gene expression analysis. Four genes (OsPAL9, OsCAD8C, OsCCR8, and OsCOMTL4) were significantly negatively correlated with lignin monomers. Furthermore, eleven genes were co-expressed in certain genotypes during secondary growth stages. Among them, six genes (OsC3H, OsCAD2, OsCCR2, OsCOMT, OsPAL2, and OsPAL8) were overlapped with microarray gene expressions, highlighting their importance in lignin biosynthesis.

Recently concluded, large-scale cancer genomics studies involving multiregion sequencing of primary tumors and paired metastases appear to indicate that many or most cancer patients have one or more “clonal" mutations in their tumors. Clonal mutations are those that are present in all of a patient’s cancer cells. Clonally mutated proteins can potentially be targeted by inhibitors or E3 ligase small molecule glues, but developing new small molecule drugs for each patient is not feasible currently. Achilles Therapeutics is currently the only company specifically targeting clonal mutations. However, they are doing so with tumor-derived T cells. To address the potential limitations of immunotherapy, I have devised another approach for exploiting clonal mutations, which I call “Oncolytic Vector Efficient Replication Contingent on Omnipresent Mutation Engagement” (OVERCOME). The ideal version of OVERCOME would likely employ a bioengineered facultative intracellular bacterium. The bacterium would initially be attenuated, but (transiently) reverse its attenuation upon clonal mutation detection.

Cell viability refers to the proportion of living, healthy cells within a given population. This information is crucial in any cell-based research, especially in toxicology. Over the past century, numerous methods for assessing cell viability have been developed. These methods can be categorized into five main types: non-invasive cell structure damage, invasive cell structure damage, cell growth, cellular metabolism, and cell membrane potential. Each method operates on different principles and has its own advantages and disadvantages, making it essential for researchers to choose the method that best fits their experimental design. Despite the variety of cell viability methods available, most rely on indirect measurements of cell viability, which may compromise their accuracy. In this sense, there is insufficient information regarding their accuracy, highlighting the need for researchers to investigate the accuracy of both existing and future methods.

Background: Lung cancer remains the leading cause of cancer-related mortality globally, predo-minantly affecting older individuals. Despite the increasing use of immune checkpoint inhibitors (ICIs) like nivolumab in non-small cell lung cancer (NSCLC), the efficacy and safety in elderly pa-tients, particularly those aged 65 and above, remain underexplored due to their underrepresenta-tion in clinical trials. Methods: This retrospective study analyzed data from 60 elderly patients (≥65 years) with metastatic NSCLC who received nivolumab as second-line or later therapy between January 2020 and May 2023. Results: The median age was 67 years, with a predominance of males (78%). Nivolumab was administered for a median of 8 cycles, with 33.3% of patients recei-ving 15 or more cycles. The median OS was 23 months, and the 1-, 3-, and 5-year survival rates were 93.3%, 54.1%, and 18.6%, respectively. Multivariate analysis identified adenocarcinoma histology, fewer than 15 cycles of nivolumab, and non-response to prior therapies as independent predictors of poor OS. Nivolumab treatment was generally well-tolerated, with 45% of patients experiencing at least grade 1 toxicity. Conclusion: Nivolumab is effective and well-tolerated in elderly patients with metastatic NSCLC, providing survival benefits comparable to those observed in younger populations. The number of treatment cycles and initial response to therapy are key determinants of survival, underscoring the importance of continued treatment in this age group.

Human Papillomavirus (HPV) related cancers account for a large percentage of cancers. The tumor microenvironment (TME) is a complex structure and contributes significantly to tumor development, progression, and invasion. The TME is particularly pronounced in HPV-related cancers and HPV is known to influence the TME. In cancer research, there is an urgent need for new models to deepen our understanding of the pathophysiology of the disease and to facilitate the development of innovative therapies. In this context, 3D cell culture has emerged as a promising method in cancer research, that offers significant potential compared to conventional approaches. 3D modelling can produce results that are more consistent with clinical outcomes without the ethical concerns and problems of animal models. In this review, our aim is to discuss the tumor microenvironment in HPV-related cancers and demonstrate the importance of advancing 3D cell culture models for this purpose.

Molten salt electrolysis stands as an indispensable technique for the production of rare earth metals, where the judicious distribution of the electric field within the electrolytic cell is paramount for ensuring the seamless operation of the electrolysis process. This study introduces a pioneering approach by integrating the Nernst-Planck equation with the Butler-Volmer equation, thereby devising a holographic electric field model tailored for the electrolysis of the NdF3-LiF molten salt system. The paper presents pioneering experimental measurements concerning the potential distribution and voltage drop characteristics within a 6kA rare earth electrolytic cell, elucidating the interplay between decomposition voltage and current density, which substantiates the model's precision. Our findings unveil that, under operational conditions of 4010 amperes, the 6kA electrolytic cell incurs a total voltage drop of 6.3 volts, delineating a melt voltage drop of 4.43 volts, an ohmic voltage drop of 0.248 volts, and an interfacial voltage drop of 1.622 volts at the electrode-electrolyte junction. The discrepancy in error is confined to within 0.3% of the actual measured voltage of 6.28 volts.The simulation outcomes from the holographic electric field model have been confirmed to precisely and comprehensively reflect the characteristics of the electric field distribution during the rare earth molten salt electrolysis process, offering valuable theoretical insights for practical manufacturing applications.

Cell-free DNA (cfDNA) analysis is a pivotal tool in non-invasive diagnostics, including cancer monitoring and prenatal testing. However, the preanalytical phase, particularly the choice of anticoagulant, significantly impacts cfDNA integrity and yield. This study aims to compare cfDNA yield, stability, and DNase activity across plasma-citrate, plasma-heparin, plasma-EDTA, and serum, to identify the most suitable specimen for cfDNA analysis among the tested anticoagulants. Blood samples from 15 healthy volunteers were collected in four types of tubes (citrate, heparin, EDTA, and serum). cfDNA was extracted and quantified using qPCR, and endogenous DNase activity was assessed through hydrolysis probe assays. Samples were incubated at 37°C for 24 hours to evaluate cfDNA degradation rates. Statistical analyses were performed using repeated measures ANOVA and Sidak's multiple comparison tests. Baseline cfDNA levels were highest in serum, followed by heparin-plasma, while EDTA and citrate-plasma exhibited the lowest levels. DNase activity was fully inhibited in EDTA-plasma, partially inhibited in citrate-plasma, and active in both heparin-plasma and serum. cfDNA degradation was most pronounced in heparin-plasma (85.3%) and serum (55.6%), compared to minimal degradation in EDTA (8%) and citrate-plasma (13.3%). Among the tested anticoagulants, EDTA-plasma proved to be the most suitable specimen for cfDNA analysis due to its low DNase activity and minimal gDNA contamination. Citrate-plasma, with intermediate DNase inhibition, could be an alternative if the citrate concentration were increased. Heparin-plasma and serum showed significant cfDNA degradation, rendering them unsuitable for cfDNA analysis. These findings highlight the importance of anticoagulant selection in preserving cfDNA integrity for diagnostic applications.

The global incidence of cutaneous squamous cell carcinoma (cSCC), a prevalent and aggressive skin cancer, has risen significantly, posing a substantial public health challenge. This study investigates the tumor microenvironment (TME) of cSCC by focusing on the spatial distribution patterns of immune and vascular markers (CD31, CD20, CD4, and CD8) using fractal dimension (FD) analysis. Our analysis encompassed 141 cases, including 100 invasive cSCC and 41 specimens with pre-invasive lesions exclusively, and the rest being peripheral pre-invasive lesions from the invasive cSCC class. The FD values for each marker were computed and compared between pre-invasive and invasive lesion classes. The results revealed significant differences in FD values between the two classes for CD20 and CD31 markers, suggesting distinct alterations in B cell distribution and angiogenic activity during cSCC progression. However, CD4 and CD8 markers did not exhibit significant changes individually. Still, the CD4/CD8 ratio showed a significant difference, indicating a potential shift in the immune landscape as lesions progress from pre-invasive to invasive stages. These findings underscore the complexity and heterogeneity of the TME in cSCC and highlight the potential of FD analysis as a quantitative tool for characterizing tumor progression. Further research is needed to elucidate the implications of these differences in the clinical management of cSCC.

Malignant lymphomas, encompassing Hodgkin lymphomas (HL) and non-Hodgkin lymphomas (NHL), represent a significant category of hematological malignancies with diverse presentations and prognoses. This review synthesizes current understanding of malignant lymphomas, focusing on their classification, incidence, epidemiology, staging, histopathology, and evolving treatment strategies. Malignant lymphomas are broadly classified into HL and NHL, with NHL further subdivided into B-cell, T-cell, and NK-cell neoplasms. The emergence of grey zone lymphomas, which exhibit overlapping features and heightened aggressiveness, underscores the complexity of diagnosis and treatment. Epidemiologically, HL predominantly affects younger individuals, while NHL incidence increases with age, reflecting a global rise in both incidence and mortality rates. Advances in diagnostic techniques and treatments have led to improved outcomes, yet challenges persist, particularly in distinguishing between overlapping lymphoma types and managing increasing incidence rates. The management of HL and NHL involves a variety of treatment options tailored to the disease stage and individual patient characteristics. After diagnosis and staging, treatment regimens may include radiotherapy, chemotherapy (either alone or in combination), and immunotherapy. Monitoring patient response is crucial, with adjustments made in cases of therapeutic failure. Achieving remission—either partial or complete—depends on the normalization of biomarkers and imaging results, reduction of lymphoid organ size, and resolution of symptoms. Despite therapeutic advances, treatment complications, including short-term effects like myelosuppression and long-term issues such as cardiomyopathy and secondary cancers, remain significant. The future of HL and NHL treatment lies in refining these approaches, minimizing adverse effects, and enhancing efficacy through innovative therapies and personalized medicine strategies.

Knee joint injuries, including those affecting the anterior cruciate ligament (ACL), meniscus, and cartilage, present complex challenges in sports medicine and orthopedics due to the intricate cellular and molecular mechanisms involved in tissue damage and repair. Understanding the molecular biology underpinning these processes is crucial for developing effective therapeutic and rehabilitation strategies. This systematic review investigates the impact of mechanical loading on the cellular responses during knee joint injury repair, with a particular focus on the molecular pathways involved in tissue regeneration. Mechanical loading plays a dual role, where controlled early loading can promote tissue repair, while excessive or inappropriate loading can exacerbate tissue damage. Fibroblasts, chondrocytes, and mesenchymal stem cells (MSCs) are central to the repair process, and their activation, proliferation, and differentiation are regulated by key molecular pathways. Upon injury, mechanotransduction pathways such as the integrin/FAK signaling axis are activated, which convert mechanical signals into biochemical responses that regulate cell adhesion, migration, and extracellular matrix (ECM) synthesis. Additionally, mechanosensitive ion channels like Piezo1 and TRPV4 modulate intracellular calcium levels, triggering downstream signaling cascades such as calmodulin/CaMKII, which regulate gene transcription and cellular responses to mechanical stress. The YAP/TAZ pathway, a critical component of the Hippo signaling pathway, responds to mechanical stimuli and regulates cell proliferation and ECM production in fibroblasts and chondrocytes. YAP/TAZ translocate to the nucleus in response to mechanical loading, where they interact with transcription factors such as TEAD, promoting the expression of genes involved in collagen synthesis and tissue repair. In parallel, growth factors like transforming growth factor-beta (TGF-β) and fibroblast growth factor (FGF) activate the TGF-β/Smad and PI3K/Akt signaling pathways, driving MSC differentiation into fibroblasts and chondrocytes, essential for ligament and cartilage repair. Mechanical loading also influences the inflammatory response at the injury site by modulating immune cell activity. Early mechanical loading can shift macrophages from a pro-inflammatory M1 phenotype to an anti-inflammatory M2 phenotype, mediated by growth factors such as TGF-β and interleukin-10 (IL-10). This phenotypic switch promotes tissue regeneration by enhancing ECM synthesis and resolving inflammation, which is crucial for long-term joint health. Matrix metalloproteinases (MMPs), regulated by NF-κB and MAPK pathways, play a role in ECM remodeling, where an imbalance between MMPs and their inhibitors (TIMPs) can lead to excessive matrix degradation, impeding tissue repair. Furthermore, Wnt/β-catenin signaling is activated in response to moderate mechanical loading, promoting chondrocyte proliferation and enhancing cartilage repair by upregulating type II collagen and aggrecan synthesis. However, dysregulation of Wnt signaling under excessive mechanical stress can lead to chondrocyte hypertrophy and cartilage degradation, contributing to osteoarthritis development. This review also explores emerging therapeutic strategies that leverage the molecular biology of knee joint repair, including biologics like platelet-rich plasma (PRP) and MSC-derived exosomes, which deliver bioactive molecules that activate critical regenerative pathways such as TGF-β/Smad and PI3K/Akt. Gene therapies targeting Wnt signaling or YAP/TAZ offer potential for enhancing tissue regeneration by modulating mechanotransduction and repair processes at the molecular level. In conclusion, the molecular biology of cellular responses to mechanical loading is central to knee joint repair following injuries. By understanding these processes and targeting specific molecular pathways, clinicians can optimize rehabilitation protocols and develop novel therapeutic approaches that enhance tissue regeneration, prevent chronic degeneration, and restore joint function. This comprehensive synthesis highlights the importance of integrating molecular insights into treatment strategies for ACL, meniscal, and cartilage injuries to improve patient outcomes.

(1) Background: Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a lifesaving treatment but carries a high infection risk. Diagnosing infections remains challenging due to the limited accuracy of standard biomarkers. (2) Methods: This single-center study aimed to evaluate presepsin (PSP) and YKL-40 as infection biomarkers in febrile patients during the allo-HSCT pre-engraftment phase. Biomarker levels were prospectively measured in 61 febrile episodes from 54 allo-HSCT patients at admission, representing baseline levels and then at Day 1, 3, 5, and 7 following fever onset. The diagnostic value was compared to that of procalcitonin (PCT). (3) Results: PSP showed fair diagnostic value on Day 1 (AUC 0.656; 95% CI: 0.510 – 0.802) and Day 3 (AUC 0.698; 95% CI: 0.559 – 0.837). YKL-40 did not provide any significant diagnostic value across measured time points. PCT outperformed PSP and YKL-40, particularly on Day 3 (AUC 0.712; 95% CI: 0.572 – 0.852). When combining biomarkers, the best model for predicting infection used PSP > 3.144 ng/mL and PCT > 0.28 g/L on Day 3, resulting in R2 of about 31% (p < 0.001). (4) Conclusion: Neither test showed sufficient discriminative power for early infection to recommend their use as individual diagnostic tools in clinical practice.

Biosensors have emerged as a promising tool for early detection of oral squamous cell carcinoma (OSCC) due to their rapid, sensitive, and specific detection of cancer biomarkers. Saliva is a non-invasive and easy-to-obtain biofluid that contains various biomarkers of OSCC, including carcinoembryonic antigen (CEA). In this study, an electrochemical biosensor for the detection of CEA in Saliva has been developed towards the diagnosis and early screening of OSCC. This biosensor utilized CEA-sensitive aptamer as sensitive elements. Fluorine-doped Tin Oxide (FTO) chip with surface modification of zinc oxide nanorod was employed as transducer. Electrochemical measurements were carried out to detect the responsive signals originated from the specific binding between aptamers and CEAs. The measurement results indicated that this biosensor was responsive to different concentrations of CEA ranging from 1 ng/mL to 80 ng/mL in a linear relationship. The limit of detection (LOD) was 0.75 ng/mL. This biosensor also showed very good specificity and regenerative capability. Stability testing over a 12-day period showed excellent performance of this biosensor. All the results demonstrated that this biosensor have great potential to be used for the detection of CEA in the saliva of OSCC patients. This biosensor provides a promising method for the rapid detection of CEA with convenience, which have great potential to be used as a new method for clinical diagnosis and early screening of OSCC.

Introduction Allogeneic stem cell transplantation (allo-SCT) has been used less frequently as a treatment modality in patients with MM. Allo-SCT may have the potential of long-term survival or even cure due to graft versus myeloma effect, however, its use continues to remain controversial given the toxicity profile related to infectious complications and graft versus host disease (GVHD). Concern for exacerbation of GVHD has been raised following re-infusion of CAR-T cells in patients with prior Allo-SCT. Ciltacabtagene autoleucel (Cilta-cel) and Idecabtagene vicleucel (Ide-cel) are two engineered CAR-T products targeting specific B-cell receptors that are approved by the FDA for the treatment of relapsed/refractory (R/R) MM. A recent retrospective study conducted by Htut et al, using data from CARTITUDE-1 trial, showed favorable efficacy and safety in patients who received Cilta-Cel with prior allo-SCT. We report our experience for patients with prior Allo-SCT who underwent therapy with CAR-T in the real world setting. Objective To evaluate the safety and efficacy of CAR-T therapy in patients with R/R MM with a history of allo-SCT. Methods We conducted a retrospective chart review of adult patients (age 18-70 years old) with R/R MM who had received CAR-T on an institutional IRB approved protocol. We collected data for the eligible patients from our institution's database. Safety outcomes included the incidence and severity of adverse events (AEs). AEs were graded using National Cancer Institute Common Terminology Criteria for Adverse Events version 5.0 (NCI-CTCAE 5.0). Cytokine release syndrome (CRS) and immune effector associated neurotoxicity syndrome (ICANS) were graded as per the American Society for Transplantation and Cellular Therapy criteria. Efficacy outcomes included overall response rates (ORR), progression free survival (PFS). Survival analysis was performed using the Kaplan-Meir method including PFS. PFS was defined as time from initiation of treatment to progression of disease or death whichever occurred first. The IMW response criteria were utilized for response assessment. Results 8 out of 56 patients who received CAR-T (Cilta-cel or Ide-cel) had a prior allo-SCT. Pts with prior allo-SCT had received a median of 7 prior lines of therapy(LOT) compared with 5 prior LOT for pts who did not(non-Allo)(p=0.04). Median time of CAR-T infusion was 98.8 months following allo-SCT. 7 of 8 (85.7%) pts experienced CRS (any grade) in Allo-SCT vs. 37 of 48 pts (77.1%) in non-Allo (p=0.48); 1 pt with prior allo-SCT experienced HLH requiring intervention with anakinra. At a median follow-up of 4.8 months, 7 of 8 patients had a response (ORR= 87.5%) in allo-SCT vs. 75%(36 of 48) in non-Allo (p=0.4), mPFS was not reached in allo-SCT vs. 11.9-months in non-Allo (p=0.5), although f/u for our cohort was short. No TRM was observed in either group. None of the patients with prior allo-SCT experienced acute GVHD following CAR-T therapy. Baseline characteristics, safety and efficacy data are summarized in Table 1. Conclusion Our findings support the conclusion that prior Allo-SCT does not impact the safety and efficacy of CAR-T therapy in patients with R/R MM. More clinical studies with a higher number of subjects and longer duration of follow-up are warranted to further evaluate the impact of Allo-SCT on CAR-T therapy's safety and clinical outcome in patients with MM.

Scientific knowledge evolves in small steps, with occasional backsteps to correct inaccuracies, all occurring within a competitive environment. This perspective for the first time looks at the history of latency-related RNA (LR-RNA) that was later renamed latency-associated transcript (LAT). At the 1986 International Herpesvirus Workshop (IHW) meeting in Leeds, England, Daniel L Rock and Anthony B Nesburn first reported the discovery of human herpes virus 1 (HSV-1) latency-related (LR)-RNA that is antisense to ICP0. Less than a month after the IHW meeting a paper was submitted to Science magazine and 8 months later appeared in print thanking “D. Rock for suggesting RNA complementary to the ICP0 message may be present in latently infected cells”. This perspective is not a review of LAT literature but intends to clarify the timeline of LAT discovery and subsequent breakthroughs such as reactivation, apoptosis, CD8+ T cell exhaustion, and LAT expression in different cell types detected during latency. While many review articles have been written about LAT since 1987, the most comprehensive and balanced review about LAT was written by Dr. David Bloom’s group (1). In this overview, I will discuss our original collaboration with Dr. Dan Rock and subsequent work that our group performed, which is still ongoing. Finally, I will discuss the controversies associated with LAT from its inception to current times.

Background: Patients with mutations in the monocarboxylate transporter 8 (MCT8) suffer from Allan-Herndon-Dudley syndrome (AHDS), characterized by developmental delay and a highly disabling movement disorder. Despite the potential of thyroid hormone derivatives to overcome the transporter defect, current trials did not achieve patient-oriented therapeutic goals. Objectives: Since most neurological symptoms are related to the dopaminergic system, we investigated the role of dopamine and its metabolites in MCT8 deficiency with regard to pathophysiology and potential therapeutic strategies in an observational cohort study. Methods: We present longitudinal data from the DEEPTYPE registry of ten patients with video-documentation, standardized phenotyping, cerebrospinal fluid analysis, treatment response to levodopa/carbidopa supplementation, and neuroimaging data. To establish a cell-based model for pathophysiological studies, we differentiated healthy human induced pluripotent stem cells (hiPSCs) into dopaminergic neurons. Results: Children presented with signs of parkinsonism in childhood, including hypokinesia, hypomimia, inability to sit or stand, rigidity, dystonia, and autonomic dysfunction along the classification of Leuzzi and colleagues. Cerebrospinal fluid homovanillic acid concentrations were decreased (n=11), suggesting isolated dopamine pathway impairment. Six out of seven patients responded favorably to levodopa/carbidopa supplementation and we did not see any adverse drug reactions. Our cell-based studies showed that hiPSC-derived dopaminergic neurons expressed MCT8 and produced quantifiable levels of biogenic amines. Conclusions: Parkinsonism is part of AHDS's clinical presentation and may be amenable to treatment. The precise impact of MCT8 deficiency on the dopamine metabolism needs to be further elucidated, e.g. by using patient iPSC-derived dopaminergic neurons in future studies.

The selection of the best allogeneic donor for each specific patient is crucial for the success of allogeneic hematopoietic stem cell transplantation (HSCT). Besides the well-known patient’s characteristics, donor-related variables such as patient-donor histocompatibility and donor age are of utmost importance for the outcome after HSCT. However, there is still debate on the definition of the best stem cell donor when multiple suitable donors exist. By using own data from two transplant centers, we here developed a calculator able to provide the 2-year patients’ overall survival (OS) associated to each of the potential donor options during the selection process. Data on n=737 HSCTs performed from January 2010 to July 2022 have been retrospectively obtained. Donor types were HLA-identical siblings, unrelated or related haploidentical. Cox regression and parametric model indicates patients’ age, disease, comorbidity index and donor type as significant variables (p<0.05) able to predict 2-year OS with robustness (concordance index: 0.677). Estimates are provided with the three donor options for each single, specific patient. Of note, estimates express real data of the transplant centers and reflect their actual experience. In conclusion, we here present the prototype of a new tool for the calculation of patients’ outcome according to the stem cell donor type, guiding transplant physicians during the process of donor selection before HSCT. The novelty of this approach relies on the use of real data from the investigating centers, reflecting the local clinical experience. Improvements are planned by filling additional information and pooling data from more centers.

The COVID-19 pandemic caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus has exposed the vulnerabilities and unpreparedness of the global healthcare system in dealing with emerging zoonoses. In the past two decades, coronaviruses (CoV) have been responsible for three major viral outbreaks, and the likelihood of future outbreaks caused by these viruses is high and nearly inevitable. Therefore, effective prophylactic universal vaccines targeting multiple circulating and emerging coronavirus strains are warranted. This study utilized an immunoinformatic approach to identify evolutionarily conserved CD4+ (HTL) and CD8+ (CTL) T cells, and B-cell epitopes in the coronaviral spike (S) glycoprotein. A total of 132 epitopes were identified, with the majority of them found conserved in the bat CoV, pangolins CoV, endemic coronaviruses, SARS-CoV-2, and Middle East respiratory syndrome coronavirus (MERS-CoV). Their peptide sequences were then aligned and assembled to identify the overlapping regions. Eventually, two major peptide assemblies were derived based on their promising immune-stimulating properties. In this light, they can serve as lead candidates for universal coronavirus vaccine development, particularly in the search for pan-coronavirus multi-epitope universal vaccines that can confer protection against current and novel coronaviruses.

Cyclin-dependent kinase 1 (Cdk1) is a master regulator of the G2-M transition between DNA replication and cell division. This study investigates the regulation of cardiomyocyte (CM) proliferation during the early neonatal period and following ischemic injury in adult mice. We analyzed cell cycle dynamics with the assessment of DNA synthesis, and cytokinesis in murine hearts during the first 15 days after birth. A distinct proliferative block was observed at 1 day, followed by a second wave of DNA synthesis at 4 days, leading to CM binucleation (CMBN) by day 5. Genome-wide mRNA profiling revealed differential expression of cell cycle regulatory genes during this period, with a downregulation of factors involved in cell division and mitosis. The loss of Cdk1 impaired CMBN but extended the neonatal CM proliferation window until day 10 post-birth. In adult hearts, cardiac-specific ablation of Cdk1 triggered CM proliferation post-myocardial infarction (MI) in specific zones, driven by the activation of EGFR1 signaling and suppression of the anti-proliferative p38 and p53 signalling. This was accompanied by restoration of fractional shortening, mitochondrial function, and decreased reactive oxygen species. Additionally, cardiac hypertrophy was mitigated, and survival rates post-MI were increased in Cdk1 knockout mice. These findings reveal a novel role of Cdk1 in regulating cell cycle exit and re-entry in differentiated CM and offer insights into potential strategies for cardiac repair.

This comprehensive overview of the historical milestones in cell culture underscores key breakthroughs that have shaped the field over time. It begins with Wilhelm Roux's seminal experiments in the 1880s, followed by the pioneering efforts of Ross Granville Harrison, who initiated groundbreaking experiments that fundamentally shaped the landscape of cell culture in the early 20th century. Carrel's influential contributions, notably the immortalization of chicken heart cells, mark a significant advancement in cell culture techniques. Holtfreter, Moscona, and Leighton introduced innovations such as 3D cell culture, laying the groundwork for further exploration. The advent of induced pluripotent stem cells by Takahashi and Yamanaka in 2006 was revolutionary, enabling the reprogramming of differentiated cells into a pluripotent state. Since then, recent innovations include spheroids, organoids, and organ-on-a-chip technologies, aiming to mimic the structure and function of tissues and organs in vitro, pushing the boundaries of biological modeling and disease understanding. In this review, we overview the history of cell culture shedding light on the main discoveries, pitfalls and hurdles that were overcome during the transition from 2D to 3D cell culture techniques. Finally, we discussed the future directions for cell culture research that may accelerate the development of more effective and personalized treatments.

Hydroperoxides (ROOH) are known as damaging agents capable of mediating mutation, while a role as signaling agents through oxidation of protein sulfhydryls that can alter cancer-related pathways has gained traction. Glutathione peroxidase 2 (GPX2) is an antioxidant enzyme that reduces ROOH at the expense of glutathione (GSH). GPX2 is noted for a tendency of large in-creases or decreases in expression levels during tumorigenesis that leads to investigators focusing on its role in cancer. However, GPX2 is only one component of multiple enzyme families that metabolize ROOH and GPX2 levels are often very low in the context of these other ROOH reducing activities. Colorectal cancer (CRC) was selected as a case study for examining GPX2 function as colorectal tissues and cancers are sites where GPX2 is highly expressed. A case can be made for a significant impact of changes in expression levels. There is also a link between GPX2 and NADPH oxidase 1 (NOX1) from earlier studies that is seldom addressed and is discussed, presenting data on a unique association in colon and CRC. Tumor-derived cell lines are quite commonly used for pre-clinical studies involving the role of GPX2 in CRC. Generally, selection for this type of work is limited to identifying cell lines based on high and low GPX2 expression with the standard re-search scheme of overexpression in low expressing lines and suppression in high expressing lines to identify impacted pathways. This overlooks CRC subtypes among cell lines involving a wide range of gene expression profiles and a variety of driver mutation differences along with a large difference in GPX2 expression levels. A trend for low and high GPX2 expressing cell lines to segregate into different CRC subclasses, indicated in this report, suggests that choices based solely on GPX2 levels may provide misleading and conflicting results by disregarding other properties of cell lines and failing to factor in differences in potential protein targets of ROOH. CRC and cell line classification schemes are presented here that were intended to assist workers in performing pre-clinical studies but are largely unnoted in work on GPX2 and CRC. Studies are often initiated on the premise that the transition from normal to CRC is associated with up-regulation of GPX2. This is probably correct. However, the source normal cells for CRC could be almost any colon cell type, some with very high GPX2 levels. These factors are addressed in this study.

According to evolutionary cancer cell biology ECCB, cancer stem cells do not arise from normal stem cells, but rather from the functional ACD phenotype of the germline. This phenotype proliferates through asymmetric cell division, producing self-renewing cells and non-proliferating daughter cells with cancer stem cell qualities (CSCs). ECCB posits that cancer stem cells themselves do not proliferate. Similar to protists, there is a close reciprocal relationship between both sister cells that collectively form a germ and stem cell lineage. These sister cells share functional roles: the proliferating cancer germline cells generate stem cells, while the non-proliferating cancer stem cells accumulate progenitor cells for new germline clones. ECCB distinguishes between primary CSCs, which are associated with carcinogenesis and primary tumors, and secondary CSCs, which are linked to metastases. This unicellular stem cell system is homologous to that of parasitic protists, such as amoebae. Both cancer stem cells and amoebae stem cells are generated by an oxygen-sensitive germline and are vulnerable to damage when oxygen levels exceed 6.0% (germline hyperoxia). Oxygen concentrations above this threshold can damage the germline genome. Dysfunctional germline cells do not undergo senescence; instead, they continue to cycle through defective symmetric cell cycles However, to regain functionality, the germline genome requires repair by hyperpolyploid giant cells known as native PGCCs, which are homologous to the multinucleated genome repair structures MGRS found in protists. Native PGCCs differ in some respects from genotoxic-induced PGCCs

We have previously demonstrated that receptor-interacting serine threonine kinase 1 (RIPK1) is expressed in hair follicle and regulates hair cycle. RIPK1 inhibitors also accelerated the telogen-to-anagen transition and elongated the anagen period in the mouse model. Here, we first investigated the involvement of RIPK1 in alopecia areata (AA). mRNA and protein expression of RIPK1 was increased in the skin of AA mouse model. Single cell RNA sequencing and immunohistochemistry showed that RIPK1 is highly increased in dendritic cells (DCs) and CD8+ T cells. RIPK1 inhibitors (i.e., Necrostatin-1s and GSK2982772) delayed the onset of AA in mouse model, and reduced the DCs and CD8+ T cells in AA skin. RIPK1 inhibitors also increased the hair length in the mouse hair organ culture mimicking AA. Collectively, these results suggest that RIPK1 is involved in AA onset via modulating immune cells, and RIPK1 inhibitors could prevent AA onset.

Developing new antibiotics poses a significant challenge in the fight against antimicrobial resistance (AMR), a critical global health threat responsible for approximately 5 million deaths annually. Finding new classes of antibiotics that are safe, have acceptable pharmacokinetic properties, and are appropriately active against pathogens is a lengthy and expensive process. Therefore, high-throughput platforms are needed to screen large libraries of synthetic and natural compounds. In this review, we present bacterial cytological profiling (BCP) as a rapid, scalable, and cost-effective method for identifying the mechanisms of action of antibiotics, offering a promising tool for combating AMR and drug discovery. We present the application of BCP for different bacterial organisms and different classes of antibiotics and discuss BCP's advantages, limitations, and potential improvements. Furthermore, we highlight the studies that have utilized BCP to investigate pathogens listed in the Bacterial Priority Pathogens List 2024 and we identify the pathogens whose cytological profiles are missing. Lastly, we explore the most recent artificial intelligence and deep learning techniques that could enhance the analysis of data generated by BCP, potentially advancing our understanding of antibiotic resistance mechanisms and the discovery of novel druggable pathways.

Sinonasal carcinomas are aggressive neoplasms that present a high morbidity and mortality rate with unfavorable prognosis. This group of tumors exhibits morphological and genetic diversity. Genetic and epigenetic alterations in these neoplasms are currently targets for diagnosis and treatment. The most common type of cancer originated in the sinonasal tract are sinonasal squa-mous cell carcinomas (SNSCC), which presents different histological patterns and variable his-tological aggressiveness. A significant number of alterations have been reported in sinonasal tu-mors, including deficiencies in the SwItch/Sucrose non-fermentable (SWI/SNF) chromatin remod-eling complex. In the sinonasal tract, deficiencies of the subunits SMARCB1/INI1, SMAR-CA4/BRG1 and SMARCA2 have been noted in carcinomas, adenocarcinomas, and soft tissue tu-mors with distinctive high-grade morphology and fatal prognosis. Objective: To identify the status of the SWI/SNF complex using immunohistochemistry in sinonasal squamous cell carci-nomas and its association with morphology and survival. Methods: A total of 1582 SNSCC cases were analyzed, selecting those with high-grade malignancy morphology, which were then evalu-ated immunohistochemically for SMARCB1 and SMARCA4 proteins. Their expression was compared with the biological behavior and survival of the patients. Results: Among the SNSCC, 47% corresponded to the non-keratinizing squamous cell carcinoma (NKSCC) type with high-grade characteristics, 40% were keratinizing squamous cell carcinomas (KSCC), 9% were SMARCB1-deficient carcinomas, and 4% were SMARCA4-deficient carcinomas. Mosaic expres-sion for SMARCB1 (NKSCC 33%, KSCC 21.9%) and SMARCA4 (NKSCC 14.6%, KSCC 12.2%) was identified, showing an impact on tumor size and progression. Conclusions: We identified that the mosaic expression of INI1 in NKSCC exhibited a worse prognosis compared to intact carcinomas and was similar to SMARCB1-deficent carcinoma.

Purpose: Testicular cancer (TC) is a rare cancer but due to early age at diagnosis and excellent cure rates, there is a big cohort of survivors. Recent studies have highlighted the late side effects of treatments of TC especially cisplatin based chemotherapy. These complications make the survivorship care challenging with detrimental effects on health and prognosis of TC survivors. In this study we provide a snapshot of common late side effects in TCS and a possible care pathway with nurse-led specialised clinic. Methods: We invited TCS in one of the cancer centre in Ireland for a comprehensive screening using questionnaires, examination and blood tests. Further investigations were performed as indicated. Mortality was assessed through retrospective chart reviews. Results: We recruited 78 TCS to participate in study. 3 patients who had died in survivorship phase were assessed through retrospective chart review. Second malignant neoplasms accounted for all three mortalities. Most common conditions in TCS after 5 years of diagnosis were hypertension (40%), dyslipidaemia (55.6%), hypogonadism (~45%) and high body mass index (52%). Majority of underlying conditions were diagnosed during screening in our clinic including two cases of coronary artery disease and one case of transient ischaemic attacks. Chemotherapy and age more than 30 years at the time of TC diagnosis were associated with a higher incidence of the late side effects. Conclusions: TC is a highly curable cancer but survivorship phase is marred by a range of late side effects. As we lack clear guidelines and care pathways for survivorship, this remains a challenge for patients and healthcare workers. A nurse-led, specialised, screening clinic can improve the care and prognosis of TC survivors.

Patients with hematologic malignancies or non-malignant diseases may undergo allogeneic hematopoietic cell transplantation (HCT), which represents a potential curative treatment. However, they are still at risk of life-threatening complications, such as relapse, acute graft-versus-host disease (GvHD), and opportunistic infections. These complications are more likely if T cell reconstitution is delayed during the initial 3-4 months after HCT. Therefore, it is of clinical importance to advance early peripheral T cell expansion.The thymus is the cradle of T cell production, but it is extremely sensitive to conditioning drugs used in HCT. As a result, egress of T cells from the thymus is abrogated during the first 3-6 months after HCT. Instead, early T cell reconstitution depends on peripheral expansion of engrafted donor T cells. However, besides its established function to produce T cells, the thymus also produces thymic peptides with hormone-like activity. These molecules play an essential part in the development and nature of immune responses, and may have a role in modulating T cell expansion and function after HCT. In this review, we investigate the role of thymic peptides in shaping the dynamics of immune reconstitution early after HCT. Furthermore, we summarize current reports on clinical application of thymic peptides post-HCT and discuss their potential use in improving patient outcomes.

Calcium phosphates and manganese phosphates play a significant role in biomedical and engineering applications, and these compounds are both synergistic. These compounds could have great potential for use in different applications, particularly as precursors of biomaterials. Therefore, the aim of this study was to explore the effect of the gradual replacement of Ca2+ with Mn2+ on the phase composition and crystal structure of brushite (CaHPO4·2H2O), which could be useful for the production of Mono- and Biphasic Ca1−xMnxHPO4·nH2O Compounds. In order to prepare the starting solutions required for this study, Ca(NO3)2·4H2O, (NH4)2·HPO4, and Mn(NO3)2·4H2O were used at various molar concentrations. Afterwards, the Ca1−xMnxHPO4·nH2O compounds were prepared using dissolution and precipitation methods. These compounds were characterized using XRD, SEM, FT-IR, XPS, and TGA techniques. The findings revealed that Mn substituted Ca partially at low Mn/Ca molar ratios, 0.25, and Mn doping increased the unit cell of CaHPO4·2H2O. The produced biphasic compound was composed of monoclinic CaHPO4·2H2O and nanostructured orthorhombic MnHPO4·(H2O)3 after a continuous increase in the Mn/Ca molar ratio to 1.5. When the Ca/Mn molar ratio was above 1.5, a monophasic compound of the MnHPO4·(H2O)3 phase was formed. The MnHPO4·(H2O)3 compound was successfully prepared for the first time using this dissolution-precipitation method. It was also established that the Mn/Ca ratio of the starting solution can be modified to produce CaHPO4·2H2O and MnHPO4·(H2O)3 phases with specific weight percent compositions.

Patients with heart failure (HF) with improved LVEF (HFimpEF) demonstrate better clinical outcomes when compared with individuals without restoration of cardiac function. Identification of predictors for HFimpEF may play a crucial role in individual management of HF with reduced EF (HFrEF). Cell-free nuclear (cf-nDNA) DNA are released from damaged cells and contribute to adverse cardiac remodeling, cardiac dysfunction and inflammation. The purpose of the study was to elucidate whether cf-nDNAs are associated with HFimpEF. It has been prescreened 1416 patients with HF using local database. Between October 2021 and August 2022 we included 452 patients chronic HFrEF after prescription of optimal guideline-based therapy and identified HFimpEF in 177 individuals from them according to the criteria of European Society of Cardiology. Measurements of circulating biomarkers were performed at baseline and in 6 months. Detection of cf-nDNA was executed with real time quantitative PCR (qPCR) using NADH dehydrogenase, ND2 and beta-2-microglobulin. We found that HFimpEF was associated with a significant decrease in the levels of cf-nDNA when compared with the patients from persistent HFrEF cohort. Presence of ischemia-induced cardiomyopathy (odds ration [OR] = 0.75; confidence interval [CI] = 0.62 – 0.88; p = 0.044), type 2 diabetes mellitus (OR = 0.77; CI = 0.71 – 0.82; p = 0.042) and digoxin administration (OR = 0.85; CI = 0.72-0.97; p = 0.042) were negative factors for HFimpEF, whereas NT-proBNP ≤1940 pmol/mL (OR = 1.42; 95% CI = 1.19-1.98, p = 0.001), relative decrease in NT-proBNP levels (&gt;35% vs. ≤35%) from baseline (OR = 1.52; 95% CI = 1.38-0.69, p = 0.001) and cf-nDNA ≤7.5 μmol/L (OR = 1.56; 95% CI = 1.07-2.94, p = 0.001) were positive predictors for HFimpEF. Multivariate logistic regression adjusted to ischemia-induced cardiomyopathy, IV HF NYHA class, and digoxin use yielded that NT-proBNP ≤1940 pmol/mL (OR = 1.43; 95% CI = 1.21-1.88, p = 0.001) and cf-nDNA ≤7.5 μmol/L (OR = 1.64; 95% CI = 1.19-2.15, p = 0.001) independently predicted HFimpEF. In conclusions: we established that the levels of cf-nDNA≤7.5 μmol/L independently predicted HFimpEF and improved discriminative ability of as single measured NT-proBNP as well as relative decrease in NT-proBNP levels ≤35% from baseline.

Background: Ubiquitination is an important post-transcriptional modification crucial for maintaining cell homeostasis. As a deubiquitination enzyme, ubiquitin-specific protease 1 (USP1) is associated with tumor progression; however, its role in bladder cancer is unknown. This study was aimed to analyze USP1 expression and study its roles in bladder cancer. Methods: The web server GEPIA was used to analyze the USP1 expression. To explore USP1 function in bladder cancer, we constructed USP1-knockout cell lines in UMUC3 cells. A FLAG-USP1 (WT USP1) plasmid and a plasmid FLAG-USP1 C90S (catalytic-inactive mutant) used to overexpress USP1 in T24 cells. CCK8, colony formation, and transwell assays were used to assess cell viability, proliferation and migration. RNA sequencing (RNA-seq) and dual-luciferase reporter assays were performed to screen the pathway. Coimmunoprecipitation and immunofluorescence were used to explore the interaction between USP1 and c-MYC. Xenograft mouse model was used to study the role of USP1 in bladder cancer. Results: USP1 expression is upregulated in human bladder cancer cells and correlated with poor patient prognosis. USP1 overexpression promoted cell proliferation, clone formation, and migration; this was attenuated by genetic ablation of USP1. Furthermore, we observed that USP1- deficiency inhibited tumor formation in vivo. Mechanistically, the c-MYC pathway was remarkably activated compared with the other pathways. Furthermore, USP1 could directly interact with c-MYC. and increase c-MYC stability depending on the catalytic activity of USP1. Conclusions: Our results suggest that high expression of USP1 promotes bladder cancer progression by stabilizing c-MYC; hence, USP1 it may serve as a novel therapeutic target for treating bladder cancer.

Conventional biochemical methods for studying cellular signaling cascades have relied on destructive cell disruption. In contrast, live cell imaging of fluorescent-tagged transfected proteins offers a non-invasive approach to understanding signal transduction events. One strategy involves monitoring the phosphorylation-dependent shuttling of a fluorescent-labeled kinase between the nucleus and cytoplasm using nuclear localization, export signals, or both. This paper introduces a simple method to visualize intracellular signal transduction in live cells by exploring the translocation properties of PKC from the cytoplasm to the membrane. We fused bait protein to PKC, allowing the bait (RFP-labeled) and target (GFP-labeled) proteins to co-translocate from the cytoplasm to the membrane. However, in non-interacting protein pairs, only the bait protein was translocated to the plasma membrane. To verify our approach, we examined the Raf–MEK–ERK signaling cascade (ERK pathway) and directly observed the Raf1/MEK2 interaction. We also identified ternary complexes containing the KSR1 scaffold protein. Additionally, the MEK/ERK interaction was dependent on the exogenous expression of KSR1.

Silver nanoparticles (AgNPs) have arisen as promising specialists in malignant growth treatment, especially in further developing radiotherapy results. In vitro examinations feature AgNPs' capacity to increment radiosensitivity in disease cell lines, prompting uplifted cell demise and decreased clonogenic endurance. Outstandingly, AgNPs display insignificant poisonousness to ordinary cells, recommending potential for specific focusing of disease cells during radiotherapy. The blend of AgNPs with cisplatin shows huge cytotoxicity and oxidative pressure, particularly in tumoral cells, underscoring the requirement for additional investigation of this therapeutic methodology. AgNPs likewise show guarantee in prompting oxidative pressure in pancreatic disease cells, recommending an expected application in pancreatic malignant growth treatment. In co-openness situations, AgNPs, when joined with cadmium, prompt metabolic variation, showing compromised cell safeguard systems. Moreover, AgNPs display potential in tweaking ABC carrier action, offering a procedure to defeat multidrug obstruction in disease cells. While featuring the significant capability of AgNPs in disease treatment, the rundown highlights the need for continuous exploration to clarify their components, well-being profiles, and clinical applications. Combining AgNPs with radiotherapy presents a promising road, tending to limits in ordinary disease therapies and underlining their multifunctionality in forming the eventual fate of malignant growth therapeutics.

To verify the efficacy of placental extracts (Pla-Ext) and human mesenchymal stem-cell-derived exosomes (hMSC-Exo) against human skin aging, we investigated the human dermal fibroblast (HDF) response following hMSC-Exo and Pla-Ext treatment and whether they elicited resistance to damage when HDFs were subjected to several types of aging stresses. The effect of hMSC-Exo alone, the combined effect of Pla-Ext and hMSC-Exo, and the effect of hMSC-Exo (Pla/MSC-Exo) recovered from cultures with Pla-Ext added to hMSC were verified using collagen, elastin, and hyaluronic acid synthase mRNA levels for each effect. Cells were subjected to photoaging (UV radiation), glycation (glycation end product stimulation), and oxidation (H2O2 stimulation) as HDF stressors. Pla-Ext did not significantly affect normal skin fibroblasts considering intracellular parameters; however, a pro-proliferative effect was observed. Pla-Ext induced resistance to several stresses in skin fibroblasts (UV irradiation, glycation stimulation, H2O2 stimulation) and inhibited reactive oxygen species accumulation following H2O2 stimulation. Although the combined efficacy of hMSC-Exo and Pla-Ext was unclear, some changes resulted in senescence stress resistance due to preconditioned hMSC-Exo stimulated by Pla-Ext. This suggests that Pla-Ext supplementation may cause some changes in the surface molecules or hMSC-Exo content (e.g., microRNA).

Polymer Electrolyte Membrane Fuel Cells (PEMFCs) have emerged as a pivotal technology in the automotive industry, significantly contributing to the reduction of greenhouse gas emissions. The high material costs of the gas diffusion layer(GDL) and bipolar plate(BP) creates a barrier for large scale commercial application. This study aims towards addressing this challenge by optimizing the material and design of the cathode, GDL and BP. While Deterministic design optimization(DDO) methods have been extensively studied, they often fall short when manufacturing uncertainties are introduced. This issue is addressed by introducing Reliability based design optimization(RBDO) to optimize four key PEMFC design variables i.e., δ_gdl, d_ch, w_ch, and w_land. The objective is to maximize cell voltage(V_cell) considering material cost of cathode gas diffusion layer(Cost_cGDL), and cathode bipolar plate(Cost_cBP) as reliability constraints. Results of the DDO show an increment in V_cell of 31mV, with reliability of around 50% for Cost_cGDL and Cost_cBP. In contrast RBDO method provides a reliability of 95% for both Cost_cGDL and Cost_cBP.

In this study, we aimed to explore the oral bacteria and fungi that can help discern oral squamous cell carcinoma (OSCC) and investigate the correlations between multiple key pathogens. Methods: Twelve participants (8 females and 4 males; mean age, 54.33 ± 20.65 years) were prospectively recruited into three groups: Group 1: healthy control, Group 2: patients with stomatitis, and Group 3: patients with OSCC, with 4 individuals in each group. Unstimulated whole saliva samples from these participants were analyzed using real-time PCR to assess the presence and abundance of 14 major oral bacterial species and Candida albicans. Results: The analysis revealed significant differences for certain microorganisms, namely Treponema denticola (T. denticola), Lactobacillus casei (L. casei), and Candida albicans. T. denticola was most abundant in the OSCC group (5,358,692.95 ± 3,540,767.33), compared to the stomatitis (123,355.54 ± 197,490.86) and healthy control (9,999.21 ± 11,998.40) groups. L. casei was undetectable in the healthy control group, but was significantly more abundant in the stomatitis group (1,653.94 ± 2,981.98) and even higher in the OSCC group (21,336.95 ± 9,258.79) (p = 0.001). A similar trend was observed for C. albicans, with DNA copy numbers rising from the healthy control (464.29 ± 716.76) to the stomatitis (1,861.30 ± 1,206.15) to the OSCC group (9,347.98 ± 5,128.54) (p = 0.006). The amount of T. denticola was positively correlated with L. casei (r=0.890, p<0.001) and C. albicans (r=0.724, p=0.008). L. casei's DNA copy number was strongly correlated with C. albicans (r=0.931, p<0.001). These three oral microbes exhibited strong positive correlations with each other and had various direct or indirect relationships with other species. Conclusions: In the OSCC group, T. denticola, L. casei, and C. albicans exhibited strong positive correlations with one another, further emphasizing the need for a deeper understanding of the complex microbial interactions in the OSCC environment.

Dendritic cells (DCs) are known as major antigen-presenting cells, and lymph nodes (LNs) play an important role in the DC-mediated immune response. This study focused on infiltration of CD1a-positive DCs (CD1a-DCs) in regional LNs in 70 cases of gallbladder cancer (GBC). The results showed that LNs infiltration by CD1a-DCs was associated with unfavorable clinical outcomes in patients with GBC in univariate analyses, but all cases categorized as CD1a-DCs high group at regional LNs had nodal metastasis. Therefore, LNs infiltration by CD1a-DCs was not an independent prognostic factor identified by multivariate analyses. The subgroup analyses in cases with LN metastasis group (n = 32), significant impact of CD1a-DCs infiltration into metastatic LNs was not observed. In contrast, CD1a-DCs infiltration into primary tumor had a significant impact on surgical outcomes. As the results of previous reports that focused on CD1a-DCs infiltration in regional LNs of other organs were varied, the role and significance of CD1a-DCs infiltration in regional LNs may be different according to the tumor histology or primary site. Accumulation of further studies is needed to clarify the role and significance of CD1a-DCs infiltration into regional LNs of solid cancers.

Although evolutionary transitions of individuality have been extensively theorized, little attention has been paid to the origin of levels of organization within organisms. How and why do specialized cells become organized into specialized tissues or organs? What spurs a transition in organizational level in cases where the function is already present in constituent cell types? We propose a model for this kind of evolutionary transition in terms of two key features of cellular self-maintenance: metabolic constraints on functional performance and the capacity for metabolic complementation between parenchymal and supporting cells. These features suggest a scenario whereby pre-existing specialized cell types are integrated into tissues when changes to the internal or external environment favor offloading metabolic burdens from a primary specialized cell type onto supporting cells. We illustrate this process of “supra-functionalization” using the nervous system and the pancreas as examples. The model reveals distinctive concerns for explaining this kind of evolutionary transition—instead of the suppression of “cheating” by components, the issue is how a tissue comes under modular genetic control as a distinct body part—and also points to a novel form of evolutionary complexity-increasing ratchet.

Background/Objectives: Hematopoietic stem cell transplantation (HSCT) in pediatric and young adult (YA) patients can lead to endotheliopathy, such as thrombotic microangiopathy (TMA), sinusoidal obstruction syndrome (SOS), and diffuse alveolar hemorrhage (DAH). Natriuretic peptides have been studied as markers of endotheliopathy and critical illness. We hypothesized that elevation in NT-proBNP was associated with development of endotheliopathy (DAH, SOS, or TMA) in the first 100 days following HSCT in pediatric and YA patients. Methods: IRB exempt status was obtained. This retrospective case-control study reviewed HSCT at our institution from 2016-2020. Cases were selected based on an endotheliopathy diagnosis in the first 100 days after HSCT. Cases were matched with controls. Baseline and near-event NT-proBNP levels were compared between cases and matched controls. Effect of NT-proBNP levels on developing endotheliopathy was estimated using conditional logistic regression. Results: Sixty-two patients were included (31 cases, 31 controls). Near-event NT-proBNP was significantly higher in cases compared to controls (Median: 473 vs. 187 pg/mL, p = 0.03, Wilcoxon rank-sum test), in contrast to comparison in baseline NT-proBNP (Median: 86 vs. 86 pg/mL, p = 0.51). After adjusting for co-variates, association between near-event NT-proBNP and odds of developing endotheliopathy did not achieve statistical significance. However, trends from most common transplant indications suggested an association between elevated near-event NT-proBNP level and endotheliopathy, particularly in acute lymphoblastic leukemia (ALL) patients. Conclusions: NT-proBNP should be studied further as a biomarker for endotheliopathy in pediatric and YA patients undergoing HSCT. This may be particularly relevant for patients undergoing HSCT for ALL.

This paper deals with the formulation of a Cobot kinematic model by developing both the direct and inverse kinematics and choosing the noa transformation matrix for defining the end-effector position and orientation with respect to the base frame. These algorithms have been validated via software in Matlab environment and by means of suitable experimental tests, which have been carried out by using a UR5e Cobot. Moreover, a specific industrial application regarding the robotized assembling of a multi-cylinder IC engine, has been designed in terms of layout and corresponding automatic cycle that is intended as a sequence of the robot elementary actions. In particular, the proposed robotized cell is composed by the UR5e Cobot along with its controller and a suitable electropneumatic circuit that includes a PLC with its user interface, which are both governed by means of a specific electronic board and a switch control box. The IC engine in scale of 1:5 and specific parts of the end-effector, which takes the role of vacuum gripper and torque limiting screwdriver, have been designed and manufactured by using a 3D printer.

Background: The implementation of tyrosine kinase inhibitors (TKIs) in the treatment of chronic myeloid leukemia (CML) has brought a significant improvement in the prognosis for CML patients and a decrease in the number of patients requiring allogeneic hematopoietic stem cell transplantation (allo-HCT). Nevertheless, the impact of TKIs on allo-HCT outcomes have not been thoroughly explored. Objectives: The main endpoint of our research was to assess the impact of prior TKI treatment on acute graft-versus-host disease (aGvHD) and chronic graft-versus-host disease (cGvHD). Methods: In our retrospective analysis we included 240 patients treated between 1993 and 2013, and divided them into 3 groups according to the therapy administered prior to matched related or matched unrelated donor allo-HCT (imatinib group n=41, dasatinib/nilotinib group n=28, TKI naïve group n=171). Results: Both the cumulative incidence of aGvHD (p=0.044) and cGvHD (p&lt;0.001) in individuals receiving second generation TKIs (2G-TKIs) prior to allo-HCT compared to patients receiving no TKIs or imatinib (IMA) (40.7% vs 61.4% vs 70.7%, p=0.044; 25.0% vs 76.4% vs 51.2%, p&lt;0.001, respectively). In case of the 2G-TKI cohort, the number of low-grade aGvHD and cGvHD was significantly lower compared to the IMA and TKI-naïve groups (p=0.018, p=0.004; p&lt;0.001 versus TKI-naïve, respectively). In terms of 3-year overall survival (OS) there were no important variations between TKI-naïve, IMA and 2G-TKI (55% vs 49.9% vs 69.6%, p=0.740). Conclusions: The results of our study suggest that TKI treatment prior to allo-HCT may have protective impact on immune-mediated outcomes.

Recent advancements in nanotechnology have led to the development of multifunctional iron oxide nanoparticles, presenting new opportunities in cancer therapy. This study aimed to synthesize and evaluate citric acid-coated/ folic acid conjugated nanoparticles loaded with doxorubi-cin and investigate their efficacy in experimental tumor models. For synthesis, controlled co-precipitation method was employed to produce highly dispersive, multifunctional nanofluids with a narrow size distribution of iron oxide nanoparticles. Nanoparticles were characterized using Dynamic Light Scattering (DLS) and Electrophoretic Light Scattering (ELS) for size distribution and zeta potential, X-ray Diffraction (XRD) and Transmission Electron Microscopy (TEM) for structure and morphology, Vibrating Sample Magnetometry (VSM) for magnetic properties, and UV-Visible (UV-Vis) and Fourier-Transform Infra-red (FTIR) spectroscopy for modification confirmation. In vitro experiments employed RM1 (prostate cancer) and MEC1 (Chronic lymphocytic leukemia) cell lines to assess cytotoxicity and drug delivery efficiency. Fluorescence microscopy confirmed the intracellular delivery of doxorubicin by these nanoparticles, highlighting their potential for targeted therapy. As a result, nanoparticles conjugated with folic acid showed reduced efficacy over time. This study underscores the critical role of nanoparticle modifications in optimizing therapeutic outcomes. Future research should focus on further refining nanoparticle formulations and long-term effects in vivo, for development of effective and safe agents for targeted cancer treatment.

The elasticity of red blood cells (RBCs) is crucial for their ability to fulfill their role in the blood. Decreased RBC deformability is associated with various pathological conditions. This study explores the application of machine learning to predict the elasticity of RBCs using both image data and detailed physical measurements derived from simulations. We simulated RBC behavior in a microfluidic channel. The simulation results provided the basis for generating data on which we applied machine learning techniques. We analyzed the surface-area-to-volume ratio of RBCs as an indicator of elasticity, employing statistical methods to differentiate between healthy and diseased RBCs. The Kolmogorov–Smirnov test confirmed significant differences between healthy and diseased RBCs, though distinctions among different types of diseased RBCs were less clear. We used decision tree models, including random forests and gradient boosting, to classify RBC elasticity based on predictors derived from simulation data. The comparison of the results with our previous work on deep neural networks shows improved classification accuracy in some scenarios. The study highlights the potential of machine learning to automate and enhance the analysis of RBC elasticity, with implications for clinical diagnostics.

Acinar cell carcinoma (ACC) accounts for about 1% of pancreatic cancers. The molecular and clinical features of ACC are less characterized than pancreatic ductal adenocarcinoma. We retrospectively evaluated the clinical and molecular features of ACC patients who underwent germline and/or somatic molecular testing at The University of Texas MD Anderson Cancer Center from 2008 to 2022. Patient information was extracted from our institutional database with the approval of Institutional Review Board. We identified 16 patients with available molecular testing results. Fourteen patients had metastatic disease, 1 had borderline resectable disease, and 1 had localized resectable disease at diagnosis. Fifteen patients were wild type for KRAS (1 patient had unknown KRAS status). Somatic/germline mutations of DNA damage repair genes (BRCA1/2, PALB2, and ATM) were present in 5 of 12 patients tested for these genes. One patient was found to have RET fusion and responded favorably to selpercatinib for over 42 months. The median overall survival (OS) was 24 months for patients with metastatic disease. One of the additional two cases which had BostonGene tests was found to have NTRK1 fusion. RNA and TME analysis by BostonGene of the two cases reported immune desert features and relatively lower RNA levels of CEACAM5, CD47, CD74 and MMP1 and higher CDH6 compared with PDAC.

Cotton fiber is the leading natural textile material, and fiber elongation plays an essential role in the formation of cotton yield and quality. Although a wealth of components in the molecular network controlling cotton fiber elongation have been reported, a lot of players still need to be functionally dissected to understand the regulatory mechanism of fiber elongation comprehensively. In the present study, an R2R3-MYB transcription factor gene, GhMYB201, was characterized and functionally verified via CRISPR/Cas9 mediated gene editing. GhMYB201 was homologous to Arabidopsis AtMYB60, and both coding genes (GhMYB201At and Dt) were preferentially expressed in elongating cotton fibers. Knocking-out of GhMYB201 significantly reduced the rate and duration of fiber elongation, resulting in shorter and coarser mature fibers. It was found that GhMYB201 could bind and activate the transcription of cell wall loosening genes (GhRDLs), and also β-ketoacyl-CoA synthase (GhKCSs) genes to enhance very-long-chain fatty acid (VLCFA) levels in elongating fibers. Taken together, our data demonstrated that the transcription factor GhMYB201s play essential roles in promoting fiber elongation via activating genes related to cell wall loosening and VLCFA biosynthesis.

Recent data has shown a continued rise in the worldwide annual incidence and mortality rates of head and neck cancers. The present standard for diagnosis and monitoring for disease recurrence or progression involves clinical examination, imaging, and invasive biopsy techniques of lesions suspected of being malignant. In addition to limitations relating to cost, time, and patient discomfort, these methodologies have inherent inaccuracies for detecting recurrence. In view of these limitations, the analysis of patient bodily fluid samples via liquid biopsy proposes a cost-effective and convenient alternative, which provides insight on the biogenetic and biomolecular underpinnings of oncologic disease processes. Liquid biopsy’s monitoring for biomarkers of head and neck cancer, including circulating tumor DNA, circulating tumor cells, and circulating cell-free RNA, has shown clinical utility in the screening, diagnosis, prognostication, and monitoring of patients with various forms of head and neck cancer. The present review will provide an update on the current literature examining the use of liquid biopsy in head and neck cancer care and the clinical applicability of potential biomarkers, with a focus on viral and non-viral circulating tumor DNA. Possible future avenues for research to address specific shortcomings of liquid biopsy will be discussed.

Type-1 Diabetes Mellitus (T1D) onset and development is closely linked with imbalance of gut microbiome, while supplementation with probiotics may result in positive alterations in gut microbiome composition. In the present study, the effect of a dietary intervention including free or immobilized cells of the presumptive probiotic Pediococcus acidilactici ORE5 on Corinthian currants, a food with beneficial impact in the condition of diabetes, was investigated. Male Wistar rats were studied which were divided into 4 groups: healthy animals, which received the free (H_FP) or the immobilized cells (H_IPC), and diabetic animals, which received the free (D_FP) or the immobilized cells (D_IPC), for 4 weeks (109 cfu/day, in all groups). At the end of the dietary intervention, D_IPC group exerted lower concentration of the inflammatory cytokine IL-1b compared to D_FP. Consumption of immobilized on Corinthian currants P. acidilactici ORE5 cells by diabetic animals led to increased loads of fecal lactobacilli and lower levels of Enterobacteriaceae, coliforms, and E. coli, while Actinobacteria phylum, Akkermansia, and Bifidobacterium genera abundances were increased, and fecal lactic acid was elevated. Overall, the results of the present research demonstrated that functional ingredients can ameliorate gut dysbiosis present in T1D and could be used to design dietary patterns aiming at T1D management. However, well-designed clinical trials are necessary, in order to confirm the beneficial effects in humans.

Because we observed a reduced humoral immune response to the COVID-19 vaccine and subsequently discovered a lower susceptibility to COVID-19 in individuals carrying the ALDH2 rs671 variant, we hypothesized that rs671 is beneficial for cellular immunity against COVID-19. Using the IFN-γ enzyme-linked immunospot (ELISPOT) assay, we evaluated cellular immunity before and after COVID-19 vaccination in two subcohorts of a previously reported cohort. Subcohort 1 (26 participants) had six repeated observations at baseline and after one to three doses, whereas subcohort 2 (19 participants) had two observations, before and after the third dose. ELISPOT counts at six months after the second dose increased from baseline in carriers of the rs671 variant, but not in non-carriers. A positive effect of rs671 on ELISPOT counts was estimated using a mixed model, including the random effect of subcohort, repeated measures, and fixed effects of vaccine type, age, sex, height, lifestyle, steroid use, and allergic disease. There was no association between ELISPOT counts and specific IgG levels, suggesting a limitation in estimating protective potential by humoral response. There is lower humoral and higher cellular immunity against SARS-CoV-2 in rs671 variant carriers compared to non-carriers, providing a potential basis for optimizing preventive measures and drug discovery.

Experiments to measure a single molecule/particle, i.e. an individual molecule/particle, at room temperature or under physiological conditions without immobilization, for example on a surface, or without significant hydrodynamic flow have so far failed. This failure has given impetus to the underlying theory of Brownian molecular motion towards its stochastics due to diffusion. Quantifying the thermodynamic jitter of molecules/particles inspires many and forms the theoretical basis of single-molecule/single-particle biophysics and biochemistry. For the first time, our simulation results for a live cell (cytoplasm) show that the tracks of individual single molecules are localized in Brownian motion, while there is fanning out in fractal diffusion (anomalous diffusion).

Experiments to measure a single molecule/particle, i.e. an individual molecule/particle, at room temperature or under physiological conditions without immobilization, for example on a surface, or without significant hydrodynamic flow have so far failed. This failure has given impetus to the underlying theory of Brownian molecular motion towards its stochastics due to diffusion. Quantifying the thermodynamic jitter of molecules/particles inspires many and forms the theoretical basis of single-molecule/single-particle biophysics and biochemistry. For the first time, our simulation results for a live cell (cytoplasm) show that the tracks of individual single molecules are localized in Brownian motion, while there is fanning out in fractal diffusion (anomalous diffusion).

Many definitions of cancer have been proposed over the years to explain cancer’s origins and pathophysiology, but the disease is too variable to allow any single description to completely encompass all of the myriad traits. A true, unifying theory of cancer must not only reveal how all known carcinogens contribute toward malignancy, but must also elucidate why cancers occur predominately in older patients, why they express so many similar behaviors, as well as account for each cancer’s individual variations. A conceptual reorganization of decades of cancer data may reveal the fundamental process that underlies the malignant transformation of cells, which not only provides a new understanding of the etiology of cancers, but also helps to interpret the subsequent events that occur after transformation, which is of significant clinical interest. The revised concepts of initiation, promotion, and progression suggest that cancer is a failed or incomplete attempt at regeneration that occurs when ordinary cellular events unintentionally coincide over time to force the use of a DNA repair pathway that is normally restricted for use in germ cells.

Diabetic peripheral neuropathy (DPN) is a prevalent complication of type 2 diabetes that affects a significant proportion of diabetic patients worldwide. Although the pathogenesis of DPN involves axonal atrophy and demyelination, the exact mechanisms remain elusive. Current research has predominantly focused on neuronal damage, overlooking the potential contributions of Schwann cells, which are the predominant glial cells in the peripheral nervous system. Schwann cells play a critical role in neurodevelopment, neurophysiology, and nerve regeneration. This review highlights the emerging understanding of the involvement of Schwann cells in DPN pathogenesis. This review explores the potential role of Schwann cell plasticity as an underlying cellular and molecular mechanism in the development of DPN. Understanding the interplay between Schwann cell plasticity and diabetes could reveal novel strategies for the treatment and management of DPN.

Recent advancements in molecular diagnostics have highlighted the potential of circulating cell-free DNA (ccfDNA) as a biomarker in various diseases. This study aims to explore the role of ccfDNA in the early diagnosis, prognosis, and therapeutic monitoring of diseases, with a focus on oncology. We conducted a comprehensive review and analysis of current literature, combined with experimental approaches involving advanced genomic sequencing and bioinformatics techniques. Our research systematically characterizes the molecular composition of ccfDNA, including its origins, size distribution, and methylation patterns, across different disease states. Furthermore, we investigate the dynamics of ccfDNA release and clearance in the bloodstream, elucidating mechanisms that could influence its diagnostic accuracy. In the context of cancer, we analyze the correlation between tumor-derived ccfDNA and disease progression, response to therapy, and overall survival. Our findings indicate that ccfDNA holds promise as a non-invasive biomarker for early disease detection, real-time monitoring of treatment efficacy, and potentially, predicting relapse. This study contributes to the growing field of liquid biopsies and opens new avenues for personalized medicine, emphasizing the need for standardized protocols in ccfDNA analysis to maximize its clinical utility. The implications of our findings extend beyond oncology, suggesting potential applications of ccfDNA in other pathological conditions.

Inflammatory bowel diseases (IBD) involve chronic inflammation of the gastrointestinal tract, where effector CD4+ T-cells play a central role. Thereby, the recruitment of T-cells into the colonic mucosa represents a key process in IBD. We recently found that CCR9 and DRD5 might form a heteromeric complex on the T-cell surface. The increase of CCL25 production and the reduction in dopamine levels associated with colonic inflammation represent a dual signal stimulating the CCR9:DRD5 heteromer, which promotes the recruitment of CD4+ T-cells into the colonic lamina propria. Here, we aimed to analysing the molecular requirements involved in the heteromer assembly as well as to determining the underlying cellular mechanisms involved in the colonic tropism given by the stimulation of the CCR9:DRD5 complex. The results show that dual stimulation of the CCR9:DRD5 heteromer potentiates the phosphorylation of the myosin light chain 2 (MLC2) and the migration speed in confined microchannels. Accordingly, disrupting the CCR9:DRD5 assembly induced a sharp reduction in the pMLC2 in vitro, decreased the migratory speed in confined microchannels and dampened the recruitment of CD4+ T-cells into the inflamed colonic mucosa. Furthermore, in silico analysis confirmed that the interface of interaction of CCR9:DRD5 is formed by the transmembrane segments 5 and 6 from each protomer. Our findings demonstrated that the CCR9:DRD5 heteromeric complex plays a fundamental role in the migration of CD4+ T-cells into the colonic mucosa upon inflammation. Thereby, disassembling the formation of the CCR9:DRD5 represents an important therapeutic opportunity to treat IBD.

The vaccines have to undergo independent testing at National Control laboratory (NCL) for various quality control tests to ensure safety, purity, and potency before market authorization and surveillance testing. In this study, five different inactivated cell culture rabies vaccine preparations (Aluminum adjuvanted and non-adjuvanted) marketed in India were evaluated using compendial safety test of mouse inoculation test (MIT) for residual live virus. Investigations were done to see the interference of aluminum adjuvant in rabies vaccine preparation to ensure safety as per the in-vivo compendial test for virus inactivation. After intracerebral injections, mice received the aluminum phosphate adjuvanted inactivated rabies vaccine (ARV) had neurological complications that resulted in their deaths between days 4 and 10. Further investigations were carried out to find out the root cause for mice death including fluorescent antibody test (FAT), rabies virus amplification test. Additionally, alamar blue based cell cytoxicity assay was carried out using BHK-21 cells to estimate the cell toxicity of aluminum phosphate adjuvanted inactivated rabies vaccine. The present study highlighted the challenges in ensuring the safety of adjuvated inactivated rabies vaccine at level of NCL. Further, study demonstrated the toxicity of aluminum under both in vitro and in vivo settings, sparking controversy regarding the safety of aluminum adjuvants in vaccines at the suggested compendial limit of 1.25 mg/single human dosage.

Comprehending the replication kinetics of SARS-CoV-2 variants helps explain why certain var-iants spread more easily and are more contagious, and pose significant health menaces to the global populations. The replication kinetics of Malaysian isolates of the Alpha, Beta, Delta, and Omicron variants were studied in the Vero E6 cell line. Their replication kinetics were deter-mined using the plaque assay, quantitative real-time PCR (qRT-PCR), and viral growth curve. The Beta variant exhibited the highest replication rate at 24 hours post-infection (h.p.i), as evi-denced by the highest viral titers and lowest viral RNA multiplication threshold. The plaque phenotypes also varied among the variants, in which the Beta and Omicron variants formed the largest and smallest plaques, respectively. All studied variants showed strong cytopathic effects after 48 h.p.i. The whole-genome sequencing highlighted cell-culture adaptation, where the Beta, Delta and Omicron variants acquired mutations at the multibasic cleavage site after three cycles of passaging. The findings suggest a strong link between the replication rates and their respec-tive transmissibility and pathogenicity. This is essential in predicting the impacts of the upcom-ing variants on the local and global populations and is useful in designing preventive measures to curb the virus outbreaks.

One of the main objectives of developing new anti-cancer vaccine strategies is to effectively induce CD8+ T cell-mediated anti-tumor immunity. Live recombinant vectors, notably Listeria mono-cytogenes, have been shown to elicit a robust in vivo CD8+ T cell response in preclinical settings. Significantly, it has been demonstrated that Listeria induces inflammatory/immunogenic cell death mechanisms such as pyroptosis and necroptosis in immune cells that favorably control immunological responses. Therefore, we postulated that the host's response to Listeria-based vec-tors and the subsequent induction of CD8+ T cell-mediated immunity would be compromised by the lack of regulatory or effector molecules involved in pyroptosis or necroptosis. To test our hy-pothesis, we used recombinant L. monocytogenes carrying the ovalbumin gene (LM.OVA) to vac-cinate wild-type (WT), caspase-1/11-/-, gsdmd-/-, ripk3-/-, and mlkl-/- C57Bl/6 mice. We performed an in vivo cytotoxicity assay to assess the efficacy of OVA-specific CD8+ T lymphocytes in eliminat-ing target cells in wild-type and genetically deficient backgrounds. Furthermore, we evaluated the specific anti-tumor immune response in mice inoculated with B16F0 and B16F0.OVA mela-noma cell lines. Our findings demonstrated that while caspase-1/11 and GSDMD deficiencies in-terfere with the rapid control LM.OVA infection, of the KO seems to contribute to the early acti-vation of OVA-specific CTL responses. In contrast, individual deficiency of each one of these pro-teins negatively impact the generation of long-lasting effector CD8+ T cells.

Introduction Image Guided Superficial Radiation Therapy (IGSRT) is a novel technology which combines traditional superficial radiation therapy (SRT) with high resolution dermal ultrasound (HRDUS) to treat non-melanoma skin cancers (NMSC). Since development, IGSRT use has expanded dramatically in outpatient clinics. We review the IGSRT literature and state of the art operating principles in US dermatologic practices. Materials and Methods A literature search of electronic databases (Medline, Pubmed, Cochrane Library, Science Direct) combined with various authors’ published and unpublished documents, procedures and clinical experience with IGSRT were synthesized for this paper. Results Studies have demonstrated IGSRT consistently delivers high cure rates( >99%) with low complications for early stage (stage 0, I, or II) squamous cell and basal cell carcinomas. Control rates are statistically superior to non-image guided SRT and external beam radiation (XRT) as well as Mohs Micrographic Surgery(MMS). This improvement is attributed to in-vivo dermal tumor visualization via HRDUS and using an interdisciplinary approach to deliver care. IGSRT use in the dermatologic clinic for early stage NMSCs has become common practice and continues to expand. Conclusion While the safety and cosmetic benefits of SRT/XRT has been long documented, IGSRT represents a significant leap forward in efficacy (statistically significant) by adding in-vivo dermal tumor imaging. Results rival and appear on one study to surpass tumor control obtained with MMS. A contributing factor to the success may be the availability and use of an interdisciplinary team approach that includes dermatologists, radiation therapists, radiation oncologists and medical physicists. The high tumor control rates, minimal side effects, favorable cosmesis and ability to treat multiple lesions per session using IGSRT is establishing this modality as a standard first line therapy for early stage NMSCs in dermatology clinics. IGSRT may represent the most effective option for the non-surgical treatment of early stage NMSC to date.

Expansins are cell wall (CW) proteins that mediate the CW loosening and regulate salt tolerance in a positive or negative way. The transcription of Populus trichocarpa expansin A6 (PtEXPA6) was downregulated upon prolonged duration of salt stress (48 h) after a transient increase induced by NaCl (100 mM). The role of PtEXPA6 in salt tolerance was determined by transferring the PtEXPA6 gene into the hybrid species Populus alba × P. tremula var. glandulosa clone 84K (84K) and Populus tremula × P. alba INRA '717-1B4' (717-1B4). The PtEXPA6-transgenic poplars of 84K and 717-1B4 showed a greater reduction in photosynthesis, stem height and diameter growth under salt stress and a reduced capacity for ROS scavenging by antioxidant enzymes, which is associated with excessive Na+ accumulation in roots and shoots. NMT flux recordings showed that the radial translocation of Na+ salt into the root xylem and real-time Na+ translocation from roots to leaves were greater in PtEXPA6-transgenic poplars than in wild-type poplars. Analysis of comparative contractility and comparative extensibility of intact root tips indicated that PtEXPA6 increased the CW loosening in the transgenic poplars of 84K and 717-1B4. Noteworthy, the PtEXPA6-promoted CW loosening was shown to facilitate radial and longitudinal transport of Na+ in transgenic poplars. We conclude that overexpression of PtEXPA6 leads to CW loosening that facilitates radial translocation of Na+ into the root xylem and subsequent Na+ translocation from roots to leaves, resulting in excessive Na+ accumulation and consequently reduced salt tolerance in transgenic poplars.

Background: Hairy cell leukemia (HCL) is a rare indolent B-cell lymphoid malignancy. The majority of patients are asymptomatic and HCL is usually diagnosed incidentally during a routine blood cell counts. In symptomatic patients, typical symptoms are related to pancytope-nia and splenomegaly. In this review we present, rare clinical symptoms in patients with HCL. Methods: A literature search was conducted of PubMed, Web of Science, and Google Scholar for articles concerning hairy cell leukemia, leukemia cutis, bone lesions, neurological manifesta-tions, pulmonary symptoms, ocular manifestations, cardiac manifestation, rare symptoms. Publications from January 1980 to August 2024 were scrutinized. Additional relevant publica-ions were obtained by reviewing the references from the chosen articles. Results: Extra medul-lary and extra nodal manifestations of classic HCL are rare. However, leukemic involvement in the skin, bone, central nervous system, gastrointestinal tract, heart, kidney, liver, lung, ocular and other organs have been reported.

The ovary is the major determinant of female reproductive health. Ovarian functions are mainly related to the primordial follicle pool, which is gradually loss with aging. Ovarian aging and reproductive dysfunctions share oxidative stress as common underlying mechanism. ROS signaling is essential for normal ovarian processes yet can contribute to various ovarian disorders when disrupted. Therefore, balance in the redox system is crucial for proper ovarian functions. In the present study, by focusing on mRNAs and ncRNAs described in the ovary and taking into account only validated ncRNA interactions, we built up an ovarian antioxidant ceRNA network, namely OvAnOx ceRNA, composed of 5 mRNAs (SOD1, SOD2, CAT, PRDX3, GR), 10 miRNAs and 5 lncRNAs (XIST, FGD5-AS1, MALAT1, NEAT1, SNHG1). Our bioinformatic analysis indicated that the components of the OvAnOx ceRNA not only contribute to antioxidant defence, but are also involved in other ovarian functions. Indeed, antioxidant enzymes encoded by mRNA of OvAnOx ceRNA operate within a regulatory network that impacts ovarian reserve, follicle dynamics, and oocyte maturation in normal and pathological conditions. The OvAnOx ceRNA represents a promising tool to unravel the complex dialogue between redox potential and ovarian signalling pathways involved in reproductive health, aging and diseases.

Measuring cell confluence in high-throughput screening of pharmaceutics, cell culture monitoring, cancer research, and industrial applications requires quantitative and standardizable measures to minimize subjective observations and increase throughput1-12 . Over the past half-century, non-invasive specialized programs have been created to quantitatively assess cell confluence, and some software tools exist to meet these needs1,4,5,6,7,8,9 . However, many of these tools are expensive, computationally-demanding, or inaccessible to non-experts1,6. With the assistance of generative artificial intelligence, we have developed the Confluence Analysis Program (CAP), a modern tool designed to be precise, interactive, and user-friendly. CAP is written entirely in Python and runs in any command-line interface (CLI), including Terminal (OS) and Command Prompt (Windows). In addition to fetching all necessary external libraries automatically and prompting drag-and-drop image retrieval, CAP features live visual comparison of cell mass identification to the original image assay. Our results demonstrate robust performance of CAP on fluorescent micrographs of varying confluences ranging from low to high. CAP is provided as a free resource, accessible in the Supplementary Materials.

Histone deacetylase inhibitors (HDACis) are being recognized as a potentially effective treatment approach for peripheral T-cell lymphoma (PTCL), which is a diverse group of aggressive lymphomas with a bleak prognosis. Recent evidence has shown that HDACi are effective in treating PTCL, especially in cases where the disease has relapsed or is resistant to other treatments, and there are few available options. Several clinical trials have demonstrated that HDACi, such as romidepsin and belinostat, can elicit long-lasting positive outcomes in individuals with PTCL, either when used alone or in conjunction with conventional chemotherapy. They exert their anti-tumor actions by regulating gene expression through inhibiting histone deacetylases, which results in the halting of cell division, programmed cell death, and the transformation of cancerous T-cell, as indicated by gene expression profile studies. Importantly, besides clinical trials, real-world evidence indicated that the utilization of HDACi presents a significant and beneficial treatment choice for PTCL. However, although HDACi showed potential effectiveness, they couldn’t cure most patients. Therefore, new combination with conventional drugs as well as new targeted agents are under investigation.

The development of a rapid, sensitive, specific method for detecting foodborne pathogens is paramount in supplying safe food to enhance public health safety. Despite the significant improvement in pathogen detection methods, key issues are still associated with rapid methods, such as distinguishing living cells from dead, pathogenic potential or health risk of the analyte at the time of consumption, detection limit, and sample-to-result. The mammalian cell-based assays analyze pathogens' interaction with the host cells and are responsive to only live pathogens or active toxins. In this study, a human embryonic kidney (HEK293) cell line expressing Toll-Like Receptor 5 (TLR-5) and chromogenic reporter system (HEK dual hTLR5) was used for the detection of viable Salmonella in a 96-well tissue culture plate. This cell line responds to low concentrations of TLR5 agonist flagellin. Stimulation of TLR5 ligand activates nuclear factor-kB (NF-κB) - linked alkaline phosphatase (AP-1) signaling cascade inducing the production of secreted embryonic alkaline phosphatase (SEAP). With the addition of a ρ-nitrophenyl phosphate as a substrate, a colored end product representing a positive signal is quantified. The assay's specificity was validated with the top 20 Salmonella enterica serovars and 19 non-Salmonella spp. The performance of the assay was also validated with spiked food samples. The total detection time (sample-to-result), including shortened pre-enrichment (4 h) and selective enrichment (4 h) steps with artificially inoculated outbreak-implicated food samples (chicken, peanut kernel, peanut butter, black pepper, mayonnaise, and peach) was 15 h when inoculated at 1–100 CFU/25 g sample. These results show the potential of HEK-DualTM hTLR5 cell-based functional biosensors for rapid screening of Salmonella.

Heat stress due to climate warming can significantly affect the synthesis of sex hormones in male adolescents, which can impair the ability of the hypothalamus to secrete gonadotropin-releasing hormone on the hypothalamic-pituitary-gonadal axis, which leads to a decrease in luteinizing hormone and follicle-stimulating hormone, which ultimately negatively affects spermatogenesis and testosterone synthesis. For optimal spermatogenesis, the testicular temperature should be 2–6°C lower than body temperature. Heat stress directly affects the testes, damaging them and re-ducing testosterone synthesis. Additionally, chronic heat stress abnormally increases the level of aromatase in Leydig cells, which increases estradiol synthesis while decreasing testosterone, lead-ing to an imbalance of sex hormones and spermatogenesis failure. Low levels of testosterone in male adolescents lead to delayed puberty and incomplete sexual maturation, negatively affect height growth and bone mineral density, and can lead to a decrease in lean body mass and an in-crease in fat mass. In order for male adolescents to acquire healthy reproductive capacity, it is recommended to provide sufficient nutrition and energy, avoid exposure to heat stress, and pro-vide foods and supplements to prevent or repair testosterone reduction, germ cell damage, and sperm count reduction caused by heat stress so that they can enter a healthy adulthood.

Treatment of HIV-associated lymphoma (HIV-Ly) with autologous stem cell transplantation (ASCT) has shown a surprisingly low relapse rate in several series. The aim of this study was to compare the clinical outcomes of HIV-Ly and lymphomas of the general population receiving ASCT. We compared two series of consecutive HIV-positive and HIV-negative patients, based on a 1:1 propensity score analysis, matching for age, sex, histology, disease status and prior therapies. We identified 44 patients in both groups. All HIV-positive patients received combination antiretroviral therapy (cART). With a median follow-up of 51 months, PFS was significantly higher in HIV-positive patients (4-years PFS 81% and 51%, in HIV-positive and HIV-negative patients, respectively, p=0.027). Four-year OS was 81% for HIV-positive and 67% in HIV-negative patients (p=0.15). The relapse rate was significantly higher in HIV-negative patients (36% vs 23%) (p=0.04). Our results clearly show that ASCT is an effective curative option for HIV-Ly, with better PFS and lower relapse rate compared to uninfected patients. A favourable effect of ASCT on HIV infection and immune system recovery, potential off-target effects of cART or other yet unknown factors may account for this observation.

(1) Background: better mechanistic understanding of desmosome disruption and acantholysis in Grover disease (GD) may improve management of this disease. Recent molecular studies highlighted promising pathways to be explored, by directly comparing GD and selected features of associated skin diseases. The association between GD and cutaneous keratinocyte carcinomas, the most prevalent non-melanoma skin cancers (NMSC), is not completely characterized. (2) Objective: To review the medical literature regarding GD-associated cutaneous keratinocyte cancers, focusing on molecular features, pathophysiological mechanisms and disease associations, to help guide future research and patient management. (3) Result: GD has been associated with a variety of skin conditions, but its association with skin cancers has been rarely reported. Between 1983 and 2024 only 9 scientific papers presented data supporting this association. Interestingly, we found that GD may mimic multiple NMSCs, as few authors reported GD cases misdiagnosed as multiple cutaneous squamous cell carcinomas for more than 4 years, or the presence of superficial basal cell carcinoma–like areas associated with focal acantholysis. (4) Conclusions: (a) GD may be an imitator of multiple NMSCs and (b) the relationship between GD and NMSCs may reveal promising pathways for the mechanistic understanding of desmosome disruption and acantholysis in GD and may even lead to its reclassification as a distinctive syndrome.

Although metastasis-associated protein 1 (MTA1) is known to play a role in cancer invasion and metastasis of various cancers, the clinical significance of its expression in canine urothelial carcinoma (UC) has not been explored. We sought to evaluate the expression of MTA1, cyclooxygenase 2 (COX2) and E-cadherin (E-cad) in association with clinicopathological parameters in clinical samples of canine UC. Here, we show that MTA1 and COX2 are overexpressed in canine UC samples compared to normal canine bladder samples, whereas E-cad levels are higher in normal bladder. The results demonstrated that MTA1 expression correlated with aggressive clinicopathological features such as high tumor-grade, muscular/vascular invasion, and metastasis. The expression of MTA1 differed in tumors depending on their localization, with the highest being in the urethra adjoining the prostate. Unexpectedly, higher E-cad levels were detected in metastatic tumor cells compared to primary tumor cells. Overall, these findings suggest that MTA1 may represent a key upstream effector tightly associated with COX2 and E-cad-mediated events in canine UC. Accordingly, MTA1 may be considered a feasible interceptive and therapeutic target for canine UC treatment.

In this paper, we report a comprehensive and consistent annotation of the locus encoding the horse T cell receptor b-chain (TRB), as inferred from the most recent genome assembly. The horse TRB locus spans approximately 1 Mb, making it the largest locus among the mammalian species stud-ied to date, with a significantly higher number of genes related to extensive duplicative events. In the region, 136 TRBV, belonging to 29 subgroups, 2 TRBD, 13 TRBJ and 2 TRBC genes, were iden-tified. The general genomic organization resembles that of other mammals, with a V-cluster of 135 TRBV genes, located upstream of two in-tandem aligned TRBD-J-C clusters, and an inverted TRBV gene at the 3’ end of the last TRBC gene. However, the horse b-chain repertoire would be affected by a high number of non-functional TRBV genes. Thus, we queried a transcriptomic da-taset derived from splenic tissue of a healthy adult horse, using each TRBJ gene as a probe to ana-lyse clonotypes encompassing the V(D)J junction. This analysis provided insights into the usage of the TRBV, TRBD and TRBJ genes and the variability of the juxtaposition region. Our results clearly demonstrated that the horse β-chain constitutes a complex level of variability broadly like that described in other mammalian species.

The key question for the understanding of corrosion phenomena of Si3N4-SiC material in Al reduction cell – whether corrosion by gases (and oxidation in particular) opens way for the corrosion by molten cryolite, or corrosion of Si3N4-SiC by cryolite plays its own role in degradation of the material in service. More probably the reactions of silicon carbide and silicon nitride with molten cryolite go after the stage of pre oxidation to silicon dioxide. The corrosion of Si3N4-SiC material in the reduction cell may take place by “gas-solid” reaction and by “liquid-solid” reaction. There are several variants of lab corrosion test for the evaluation of corrosion resistance of Si3N4-SiC material to cryolite. The results of investigation of Si3N4-SiC lab corrosion test give no direct evidence of selective dissolution of specific phase (Si3N4 or SiC, α-Si3N4 or β-Si3N4) in cryolite. The existing variants of lab corrosion testing require clarification.

The management of vision-threatening retinal diseases remains challenging due to the lack of an effective drug delivery system. Encapsulated cell therapy (ECT) offers a promising approach for the continuous delivery of therapeutic agents without the need for immunosuppressants. In this context, an injectable and terminable collagen-alginate composite (CAC) ECT gel, designed with a Tet-on pro-caspase-8 system, was developed as a safe intraocular drug delivery platform for the sustained release of glial cell line-derived neurotrophic factor (GDNF) to treat retinal degenerative diseases (RDDs). This study examined the potential clinical application of the CAC ECT gel, focusing on its safety, performance, and termination through doxycycline (Dox) administration in the eyes of healthy New Zealand White rabbits, as well as its therapeutic efficacy in rabbits with sodium iodate (SI)-induced retinal degeneration. The findings indicated that CAC ECT gel can be safely implanted without harming the retina or lens, displaying resistance to degradation, facilitating cell attachment, and secreting bioactive GDNF. Furthermore, the GDNF levels could be modulated by the number of implants. Moreover, Dox administration was effective in terminating gel function without causing retinal damage. Notably, rabbits with retinal degeneration treated with the gels exhibited significant functional recovery in both a-wave and b-wave amplitudes and showed remarkable efficacy in reducing photoreceptor apoptosis. Given its biocompatibility, mechanical stability, controlled drug release, terminability, and therapeutic effectiveness, our CAC ECT gel presents a promising therapeutic strategy for various retinal diseases in a clinical setting, eliminating the need for immunosuppressants.

The bi- or multi-nucleated Reed-Sternberg cell (RS) is the diagnostic cornerstone of EBV-positive and EBV-negative classical Hodgkin lymphoma (cHL). cHL is a germinal center (GC) derived B-cell disease. Hodgkin cells (H) are the mononuclear precursors of RS. An experimental model has to fulfill three conditions to qualify as common pathogenic denominator: i) to be of germinal center-derived B-cell origin, ii) to be EBV-negative to avoid EBV latency III expression, and iii) to support permanent LMP1 expression upon induction. These conditions are unified in the EBV-, Diffuse Large B-Cell Lymphoma (DLBCL) cell line BJAB-tTA-LMP1. 3D reconstuctive nanotechnology revealed spatial, quantititive and qualitative disturbance of telomere/shelterin interactions in mononuclear H-like cells, further progression during transition to RS-like cells, including progressive complexity of the karytotype with every mitotic cycle, due to BBF (breakage/bridge/fusion) events. The findings of this model were confirmed on diagnostic patient samples and correlate with clincal outcome. Moreover, in vitro signficant disturbance of the lamin AC/telomere interaction progressively occured. In summary, our research over the past three decades identifed cHL as the first lymphoid maligancy driven by a disturbed telomere/shelterin/lamin AC interaction, generating the diagnostic RS. Our findings may act as trailblazer for tailored therapies in refractory cHL.

Glioma is the most common primary malignant brain tumors. Despite advances in surgical techniques and treatment regimens, the therapeutic effects remain unsatisfactory. Immunotherapy has brought new hope to glioma patients, but the immunosuppressive nature of the tumor microenvironment (TME) has limited the therapeutic outcomes. This study aims to reveal the subpopulations and functional characteristics of tumor-associated macrophages (TAMs), to explore the regulatory effects of chlorogenic acid on the immune microenvironment, and investigate its potential for clinical application. In this study, Chlorogenic acid slowed tumor growth in the mouse model and extended the survival time of the mice. ScRNA-seq analysis elucidated the differentiation trajectories and functional characteristics of BMDMs and microglia, revealing the regulatory effects of chlorogenic acid on these two populations. It enhanced the antigen-presenting function of macrophages and the gene expression related to T-cell immune activation, activated microglia through the JAK-STAT pathway, and improved antitumor functions. Using the target prediction database, a PPI and molecular docking model were constructed, confirming the good affinity of chlorogenic acid with STAT1. A case of a patient treated with chlorogenic acid was reviewed, who survived for 5 years and 6 months, achieved partial remission (PR) after 9 months of treatment, and remains alive with no new symptoms or toxic side effects. Our study revealed the subtypes and differentiation trajectories of tumor-associated macrophages. Chlorogenic acid significantly improved the immune microenvironment of glioma by modulating the function of bone marrow-derived mono-macrophages (BMDMs) and microglia, demonstrating good antitumor effects. This study provides new insights for targeting the regulation of the TME and offers theoretical and practical support for the clinical application of chlorogenic acid.

During the last few years, the world witnessed one of the worst calamities to humankind- the COVID-19 pandemic. We also witnessed the development of vaccines at an unprecedented pace against the SARS-CoV-2- the causative agent. India was among the hard-hit countries by COVID-19, developed its indigenous vaccine and implemented vaccines developed in other countries as well. Initially, the antibody level and the neutralizing antibody titer against the pathogen were considered as the correlates of immune protection from the disease. However, the long-term protection from the pathogen was provided by the immune memory cells viz. T and B memory cells and their ability to efficiently make recall responses. In this regard, globally the research studies focused mainly on the mRNA-based vaccines. The studies on immune memory response, particularly- the B cell memory response bestowed by the vaccines given to Indians is relatively obscure. We studied the level of RBD specific memory B cells in the peripheral circulation of the study participants and the ability of B cells to secrete antigen-specific antibodies among Indians vaccinated with Covaxin (BBV152), Covishield (AZD1222), Corbevax (BECOV2D) and Sputnik Light (Gam-COVID-Vac) as well as unvaccinated individuals. Our study revealed that Corbevax and Sputnik Light conferred better antibody-secreting cell (ASC) responses.

Efficient beam steerable high-gain antennas enable high-speed data rates over long-distance networks, including wireless backhaul, satellite communications (SATCOM), and SATCOM On-the-Move. These characteristics are essential for advancing contemporary wireless communication networks, particularly within 5G and beyond. Various beam-steering solutions have been proposed in the literature, with passive beam-steering mechanisms employing planar metasurfaces emerging as cost-effective, power-efficient, and compact options. These attributes make them well-suited for use in confined spaces, large-scale production and widespread distribution to meet the demands of the mass market. Utilizing a dual-band antenna terminal setup is often advantageous for full duplex communication in wireless systems. Therefore, this article presents a comprehensive review of the dual-band beam steering techniques for enabling full-duplex communication in modern wireless systems, highlighting their design methodologies, scanning mechanisms, physical characteristics, and constraints. Despite the advantages of planar metasurface-based beam steering solutions, literature on dual-band beam steering antennas supporting full duplex communication is limited. This review article identifies research gaps and outlines future directions for developing economically feasible passive dual-band beam-steering solutions for mass deployment.

The immune response is crucial in Pregnancy Loss (RPL), which affects 1-2% of women. In RPL, peripheral blood lymphocytes and local immune cells are activated with a concomitant decrease in the leukocyte tolerogenic response in the uterus and decidua. IL-17 plays an essential role in T cell activation, weakening tolerogenic responses of the T cells. The report aims to analyze the IL-17 genetic polymorphism, circulating IL-17 lymphocyte populations, the reaction upon lymphocyte priming, the amount of cytokine in serum, and the number of T-reg cells in a group of 50 RPL and 50 normal women from the admixed Venezuelan population. No differences were observed in the genetic polymorphisms rs2275913 and rs763780. A significant decrease (P&lt;0.001) was observed in RPL patients compared to controls in the IL-17+ cell populations CD3+, CD8+, CD56+, and CD4CD25 FoxP3, but not in the CD4IL17+ cells. The response upon cell priming was significantly decreased (P&lt;0.001) in all subpopulations of RPL as compared to controls. It is concluded that despite the increase in IL-17A in the serum of patients, the number of circulating IL-17 cells and the lymphocyte response to stimulation was significantly decreased (p&lt;0.001). No differences in the genetic polymorphism compared to controls suggest that biological factors influence IL-17 levels in RPL patients, which must be carefully studied.

Related studies have shown that mitophagy plays a role in the progression of Atherosclerosis (AS). Utilizing GSE43292, GSE100927, and GSE159677 datasets, WGCNA and differential expression analysis identified mitophagy-related module genes and DEGs1, while single-cell analysis screened DEGs2. MR analysis revealed causal AS-linked genes in DEGs1. LASSO and expression validation identified biomarkers, evaluated via ROC curves. Further analyses included GSEA, immune infiltration, ANN, regulatory networks, and drug prediction. Four candidate genes were identified through WGCNA, differential expression analysis and MR analysis. The LASSO analysis identified CD36, IL1RN, and MME as biomarkers, while the MR analysis showed that IL1RN and MME are protective factors and CD36 is a risk factor in AS. The ROC curves disclosed that the ANN model had greater diagnostic power for AS than any other biomarker. The GSEA revealed that the biomarkers were enriched in lysosomes and hematopoietic cell lineage and were significantly positively correlated with monocytes. The regulatory network construct showed that small nucleolar RNA host gene 16 regulates MME via the micro RNAs. This study identified CD36, IL1RN, and MME as biomarkers of mitophagy in AS and indicated that they could be applied in the diagnosis and treatment of this condition.

One of the bottlenecks to bringing new therapies to the clinic has been a lack of vectors for delivering novel therapeutics in a targeted manner. Cell penetrating peptides (CPPs) have received a lot of attention and have been the subject of numerous developments since their identification nearly 3 decades ago. Known for their transduction abilities, they have generally been considered inert vectors. In this review we present a schema for their classification, highlight what is known about their mechanism of transduction, and outline the existing literature, as well as our own experience, vis a vis the intrinsic anti-inflammatory properties that certain CPPs exhibit. Given the inflammatory responses associated with viral vectors, CPPs represent a viable alternative to these vectors; further, the anti-inflammatory properties of CPPs, mostly through inhibition of the NF-κB pathway, are encouraging. Much work in relevant animal models, toxicity studies in large animal models, and ultimately human trials are needed before their potential is fully realized.

While adeno-associated viral (AAV) vectors are successfully used in a variety of in vivo gene therapy applications, they continue to be hampered by the immune system. Here, we sought to identify innate and cytokine signaling pathways that promote CD8+ T cell responses against the transgene product upon vector administration to murine skeletal muscle. Eliminating just one of several pathways (including DNA sensing via TLR9, IL-1 receptor signaling, and possibly endosomal sensing of double-stranded RNA) substantially reduced the CD8+ T cell response at lower vector doses but was surprisingly ineffective at higher doses. Using genetic, antibody-mediated, and vector engineering approaches, we show that blockade of at least two innate pathways is required to achieve an effect at higher vector doses. Concurrent blockade of IL-1R1>MyD88 and TLR9>MyD88>IFN I>IFNaR pathways was often but not always synergistic and had limited utility in preventing antibody formation against the transgene product. We further provide evidence that CpG depletion of vector genomes and inclusion of TLR9 inhibitory sequences can have a synergistic effect. However, even low-frequency CD8+ T cell responses could eliminate transgene expression. Thus, innate immune avoidance/blockade strategies, albeit helpful, may not be sufficient to prevent destructive cellular responses in muscle gene transfer because of redundancy of immune activating pathways.

The grand antiprism A is an outlier among the uniform 4-polytopes, since it is not obtainable from Wythoff’s construction. Its symmetry group G(A) has been incorrectly described as [[10,2+,10]] or even as an `ionic diminished Coxeter group’. In fact, G(A) is another group of order 400, namely the group ±[D10×D10]·2, in the notation of Conway and Smith.

During the COVID-19 pandemic, antibody-based vaccines targeting the SARS-CoV-2 spike glycoprotein were the key focus for development because neutralizing antibodies were associated with protection against the SARS-CoV-2 infection in pre-clinical models and human trials. While deploying these spike-based vaccines saved millions of lives worldwide, it has become clear that the immunological mechanisms of protection against severe disease are multifaceted and involve non-neutralizing antibody components. Here, we describe a novel pan-sarbecovirus T-cell vaccine, ChAdOx1.COVconsv12, designed to complement and broaden the protection of spike vaccines. The vaccine immunogen COVconsv12 employs the two regions in the viral proteome most conserved among sarbecoviruses delivered by replication-deficient vector ChAdOx1. It focuses T cells towards epitopes shared among sarbecoviruses including evolving SARS-CoV-2 variants. Here, we show that ChAdOx1.COVconsv12 induced broad T-cell responses in the BALB/c and C57BL/6 mice. In the Syrian hamster challenge model, ChAdOx1.COVconsv12 alone did not protect against the SARS-CoV-2 infection, but when co-administered with 1/50th of the ChAdOx1 nCoV-19 spike vaccine protective dose, faster recovery and lower oral swab viral load were observed. Induction of CD8+ T cells may decrease COVID-19 severity and extend the T-cell response coverage of variants to match the known (and as yet unknown) members of the beta-coronavirus family.

Background: Cardiac development is a complex developmental process. The early cardiac straight tube is composed of an external myocardial layer and an internal endocardial lining. Soon after rightward looping, the embryonic heart becomes externally covered by a new epithelial lining, the embryonic epicardium. A subset of these embryonic epicardial cells migrate and colonize the embryonic myocardium, contributing to distinct cell types. In recent years, our understanding of the molecular mechanisms that govern proepicardium and embryonic epicardium formation has greatly increased. We have recently witnessed a novel layer of complexity governing gene regulation with the discovery of non-coding RNAs. Our laboratory recently identified three distinct lncRNAs, adjacent to the Wt1, Bmp4 and Fgf8 chicken gene loci, with enhanced expression in the proepicardium that are distinctly regulated by Bmp, Fgf and thymosin 4, providing support of their plausible implication in epicardial formation. Methods: Expression of lncRNAs was analyzed in different chicken and mouse tissues as well as their subcellular distribution in chicken proepicardial, epicardial, ventricle explants and in different murine cardiac cell types. lncRNA transcriptional regulation was analyzed by using siRNAs and expression vectors of different transcription factors in chicken and mouse. Antisense oligonucleotides were used to inhibit Gm14014 expression. Cardiac injury was induced ex vivo in mouse ventricle explants by cryoinjury. Furthermore, RT-qPCR, immunocytochemistry, RNA pulldown, Western blot, viability and cell migration assays were conducted to investigate the biological function of Wt1_76127 and Gm14014. Results: We demonstrated that Wt1_76127 in chicken and its evolutionarily conserved homologue Gm14014 in mice are widely distributed in different embryonic and adult tissues and distinctly regulated by cardiac enriched transcription factors, particularly Mef2c and Nkx2.5. Gm14014 is distinctly regulated in mouse ventricular cryoinjury ex vivo models, displaying a negative correlation with epicardial and epithelial to mesenchymal transition markers. Furthermore, silencing assays demonstrated that mouse Gm14014, but not chicken Wt1_76127, is essential for epicardial, but not endocardial or myocardial cell migration. Such process is governed by partnering with Myl9, promoting cytoskeletal remodeling. Conclusion: Our data evidence that Gm14014 plays a pivotal role in epicardial cell migration essential for heart regeneration.

Background: Cancer stem cells (CSC) are small subpopulation of cancer cells that has the potential to self-renew and has the strong proliferative capacity that sustain tumorigenesis capability. This ability of the CSCs to escape immune responses make the CSCs a primary source of functionally altered, immune-resistant, chemo-resistant aggressive tumor cells. Mentioned characteristic determines the potential advantage of targeting CSCs for the treatment of solid tumor. Method: We downloaded different gene expression datasets of CSCs from the NCBI-GEO database and identified common gens by using suitable Venn tool. Then explored the prognostic significance of the particular genes in particular cancer along with analysis of expression of these genes at protein level in human carcinomas by using different webserver. Finally, using toxicogenomic database we selected several important drugs or chemicals. Result: In this study, we identified common targets between three types of solid tumor and the prognostic significance of the identified genes. The identified drugs capable of targeting the expression or signaling network of designated genes of CSC that may contribute in CSC targeted cancer therapy. Conclusion: We identified several CSC associated genes in human breast, lung and ovarian carcinomas and several drugs that can target the function of these genes. Our study also suggests that there is need for more experimental investigations to find out the actual functional activity and the mechanism of action of the genes of CSC.

Hemoglobin S (Hb S) is genetically inherited variant of hemoglobin affecting around 300 000 infants are born annually. Facilitates for universal population testing of carrier state are available only in developed countries. The screening tests for Hb S have their respective limitations and the confirmatory tests are costly and require experts for conduct and interpretation. The use of two tests will enhance the overall diagnostic accuracy at the level of screening. We have considered two scenarios for population screening for SCD – community based in health camp mode / house to house survey and facility-based mode at health center. The combination of lateral flow immunoassay based POCT with sickling test can be used in absence of HPLC for population screening. However, for antenatal women portable microelectrophoresis based test is recommended as it flags the presence other hemoglobin variants along with Hb S.

In this very personal hunt for the meaning of the bacterial cell cycle, the snark, I briefly revisit and update some of the mechanisms we and many others have proposed to regulate the bacterial cell cycle. These mechanisms, which include the dynamics of calcium, membranes, hyperstructures and networks, are based on physical and physico-chemical concepts such as ion condensation, phase transition, crowding, liquid crystal immiscibility, collective vibrational modes, reptation, and water availability. I draw on ideas from subjects such as the ‘prebiotic ecology’ and phenotypic diversity to help with the hunt. Given the fundamental nature of the snark, I would expect that its capture would make sense of other parts of biology. The route therefore followed by the hunt has involved trying to answer questions like “why do cells replicate their DNA?”, “why is DNA replication semi-conservative?”, “why is DNA a double helix?”, “why do cells divide?”, “is cell division a spandrel?” and “how are catabolism and anabolism balanced?”. Here, I propose some relatively unexplored, experimental approaches to testing snark-related hypotheses and, finally, I propose some possibly original ideas about DNA packing, about phase separations, and about computing with populations of virtual bacteria.

Berberine (BER) is a natural alkaloid found in various Berberis species, used traditionally in herbal medicine and dietary supplements. BER exhibited various biological activities, including anti-inflammatory, antioxidant, antidiabetic, and anti-tumour properties by modulating cellular pathways and influencing gene expression. While BER shows promise in various health applications, its possible toxicity raises concerns. In 2023, EFSA issued a call for data on BER- and protoberberine (PROTBER)-containing products, to comprehensively assess its safety and potential health claims. With these premises, new data was collected by assessing: i) the phytochemical profile of BER- and PROTBER-containing preparations by newly developed HPLC-DAD method; ii) in silico and in vitro investigation of pharmacokinetics properties of BER; iii) in vitro cytotoxicity in different human cell lines and, iv) biological and molecular activities of extracts containing BER and PROTBER in in vitro human non-cancer cells. By testing 16 commercial Berberis extracts, the evidence collected showed a good qualitative reliability in terms of phytochemical profile and limited relapses on cytotoxicity even at 200 µg/ml after a 24h of exposure, while evidencing a good cellular bioavailability and cell migration inhibition. The potential for plant-drug interactions affecting CYP450 proved to be minimal. Overall, our work provides a useful overview to better elucidate potential concerns on botanicals containing BER and minor PROTBER commonly used in food supplement.