Cell Components Research Articles

Modeling Acoustic Attenuation, Sound Velocity and Wave Propagation in Lithium‐Ion Batteries via a Transfer Matrix

AbstractA simple 1D transfer matrix model of a battery is introduced and parametrized using harvested individual cell components at 0 % and 100 % SoC. This model allows for the calculation of group velocity and attenuation. The results of the model show good agreement with measured values, highlighting increased attenuation and group velocity at the resonances. This emphasizes the importance of selecting a suitable interrogation frequency for ultrasound investigations in lithium‐ion batteries. The model accurately replicates the observed weakening of resonances with increasing SoC. Additionally, it provides the basis to fit US spectroscopy data in the future, enabling immediate determination of component thickness and the Young's modulus of individual components, along with aiding in the identification aging effects of the anode and cathode materials. The model can visualize wave propagation within the battery. At certain frequencies, standing waves form which could be used in high‐intensity ultrasound applications targeted at individual cell components.

Carvacrol alleviates LPS-induced myocardial dysfunction by inhibiting the TLR4/MyD88/NF-κB and NLRP3 inflammasome in cardiomyocytes.

Sepsis-induced myocardial dysfunction (SIMD) may contribute to the poor prognosis of septic patients. Carvacrol (2-methyl-5-isopropyl phenol), a phenolic monoterpene compound extracted from various aromatic plants and fragrance essential oils, has multiple beneficial effects such as antibacterial, anti-inflammatory, and antioxidant properties. These attributes make it potentially useful for treating many diseases. This study aims to investigate the effects of CAR on LPS-induced myocardial dysfunction and explore the underlying mechanism. H9c2 cells were stimulated with 10µg/ml LPS for 12h, and c57BL/6 mice were intraperitoneally injected with 10mg/kg LPS to establish a septic-myocardial injury model. Our results showed that CAR could improve cardiac function, significantly reduce serum levels of inflammatory cytokines (including TNF-α, IL-1β, and IL-6), decrease oxidative stress, and inhibit cardiomyocyte apoptosis in LPS-injured mice. Additionally, CAR significantly downregulated the expression of TLR4, MyD88, and NF-κB in LPS-injured mice and H9c2 cells. It also inhibited the upregulation of inflammasome components (such as NLRP3, GSDMD, and IL-1β) in H9c2 cells triggered by LPS. Taken together, CAR exhibited potential cardioprotective effects against sepsis, which may be mainly attributed to the TLR4/MyD88/NF-κB pathway and the NLRP3 inflammasome.

Interfacing Complex Coacervates with Natural Cells

Coacervates have been investigated as protocells or synthetic cells, as well as subcellular compartments for the creation of new materials, thus bridging the gap between living and non‐living systems in materials science, synthetic biology, and bioengineering. Given the design flexibility and simplicity of coacervates, along with the functionality and complexity of natural cells, the interfacing of complex coacervates with natural cells is considered significant for various biotechnological and biomedical applications. In this review, the fundamental mechanisms and underlying theories of coacervate systems are introduced. Recent efforts to interface coacervates with natural cells are summarized in three key scenarios: (i) the integration of coacervates with natural cell components for the living material assembly into protocells; (ii) communication between therapeutic synthetic cells and natural cells for drug delivery and cell repair; and (iii) the formation of intracellular condensates for metabolic regulation, followed by the regulation of their phase transitions for pathological elucidation. Finally, the potential of coacervate‐natural cell interfaces is discussed in the context of developing living/synthetic cell constructs, creating precise disease therapy strategies, and advancing programmable metabolic engineering networks.

The impact of varying sizes of silver nanoparticles on the induction of cellular damage in Klebsiella pneumoniae involving diverse mechanisms

Abstract Silver nanoparticles (AgNPs) are extensively studied as potent antibacterial agents targeting antibiotic-resistant pathogens. Cellular damage induced through various mechanisms that can affect multiple cell components like the outer membrane, enzymes, and proteins is closely linked to their chemical and morphological characteristics. We investigated the impact of AgNPs’ size on their antibacterial effectiveness using two differently sized nanoparticles: silver nanoparticle-Citrus limon (AgCL) with an average size of 21 nm and silver nanoparticle-Citrus sinensis (AgCS) with an average size of 13 nm, derived from C. limon and C. sinensis through environmentally friendly methods. The study evaluated their antibacterial effects by assessing morphology changes via scanning electron microscopy, metabolic alterations using Fourier transform infrared (FT-IR) spectroscopy, and oxidative stress responses through biochemical markers in Klebsiella pneumoniae cells exposed to AgNPs. The results showed that both AgCL and AgCS exhibited remarkable antibacterial activity, evidenced by inhibition zones of 14 ± 1.5 and 16 ± 1.0 mm, respectively. Morphological changes in K. pneumoniae cells treated with AgNPs were size dependent, with notable alterations noted. FT-IR spectroscopy revealed size and concentration-dependent biochemical changes, particularly in shifts in functional groups involved in the fluidity of cell wall lipid, and protein structure. Exposure to AgNPs led to increased oxidative stress markers like lipid peroxides and reduced levels of enzymatic and non-enzymatic antioxidants, more prominently observed with smaller AgCS nanoparticles (13 nm). AgNPs induce oxidative stress and morphological changes in K. pneumoniae strains, with smaller nanoparticles demonstrating greater efficacy. These findings underscore the importance of nanoparticle size in optimizing the antibacterial properties against pathogens.

Comprehensive Study on Cell Components in High‐Voltage Pouch Cells with Lithium Perchlorate: Decomposition, Transesterification, Chlorination, Deposition, and Self‐Discharge

AbstractBattery development has traditionally focused on high energy and long lifetime cells, but there is now a shift towards their sustainability and safety. One example of this trend is the search for fluorine‐free conductive salts. The overwhelming majority of lithium‐ion conductive salts contain fluorine, which is critical regarding their environmental impact, sustainability, and toxicology. In this study, we perform a comprehensive investigation of the performance and aging mechanisms of cell components with LiClO4 as conductive salt in high‐voltage NMC622‖Graphite pouch cells. The cells containing LiClO4 show poorer electrochemical performance compared to their LiPF6 equivalents. However, to the best of our knowledge, a mechanistic understanding of the effect of LiClO4 on the aging of electrode and electrolyte components for high‐voltage cells is largely missing. Developing such an understanding will pave the way toward designing alternative salts to LiPF6, ultimately leading to fluorine‐free and more sustainable battery cells. Our results show, that the chlorination of ethyl methyl carbonate at both methyl and ethyl groups and the formation of large (Liw)AlxOyClz composite deposits on the cathode surface result from perchlorate degradation at the cathode. This leads to increased cell resistance, reduced capacity retention, and accelerated degradation of the LiClO4‐containing electrolytes.

Improved identification of clinically relevant Acute Leukemia subtypes using standardized EuroFlow panels versus non-standardized approach.

Rare acute leukemia (AL) components or subtypes such as blastic plasmacytoid dendritic cell neoplasm (BPDCN) or early T-cell precursor acute Lymphoblastic Leukemia (ETP-ALL) can be difficult to detect by routine flow cytometry due to their immunophenotypes overlapping with other poorly differentiated AL. We hypothesized that using standardized EuroFlow™ Consortium approach could better diagnose such entities among cases that previously classified as acute myeloid leukemia (AML)-M0, AML with minimal differentiation, AML with myelodysplasia-related changes without further lineage differentiation, and AL of ambiguous lineage. In order to confirm this hypothesis and assess whether these AL subtypes such as BPDCN and ETP-ALL had previously gone undetected, we reanalyzed 49 banked cryopreserved sample cases using standardized EuroFlow™ Consortium panels. We also performed target sequencing to capture the mutational commonalities between these AL subtypes. Reanalysis led to revised or refined diagnoses for 23 cases (47%). Of these, five diagnoses were modified, uncovering 3 ETP-ALL and 2 typical BPDCN cases. In 12 AML cases, a variable proportion of immature plasmacytoid dendritic cell and/or monocytic component was newly identified. In one AML case, we have identified a megakaryoblastic differentiation. Finally, in five acute lymphoblastic leukemia (ALL) cases, we were able to more precisely determine the maturation stage. The application of standardized EuroFlow flow cytometry immunophenotyping improves the diagnostic accuracy of ALs and could impact treatment decisions.

Toward revolutionizing PEMFC manufacturing for clean energy conversion: a review on the innovative contribution of 3D printing techniques

AbstractThree-dimensional printing (3DP) is a technology useful for fabricating both structural and energy devices. Of great concern to this review is promising nature of additive manufacturing (AM) for engineering fuel cells (FCs) for clean energy conversion. 3DP technique is useful for the fabrication of fuel cell components, and they offer waste minimization, low-cost, and complex geometric structures. In this review, significance of different 3DP techniques toward revolutionizing fuel cell fabrication is given. The aim is to unravel the importance and status of 3D-printed fuel cells and hence provides researchers and scientists with extensive opportunities of 3DP techniques for fuel cell engineering. After careful selection of state-of-the-art literatures, different kinds of 3DP techniques of relevance to electrolytes, electrodes, and other key components (e.g., gas diffusion layers (GDLs), bipolar plates (BPs), and membrane electrode assembly (MEA)) fabrication are explicitly discussed. Among the techniques, the best approaches are recommended for further studies. Advantages associated with these techniques are indicated for the benefit of those whose interests matter most on clean energy production. The challenges researchers are facing in the use of 3DP for fuel cell fabrications are identified. Possible solutions to the identified challenges are suggested as way forward to further development in this research area. It is expected that this review article will benefit engineers and scientists who have interest on clean energy conversion devices.

Cancer cells sense solid stress to enhance metastasis by CKAP4 phase separation-mediated microtubule branching.

Solid stress, originating from rigid and elastic components of extracellular matrix and cells, is a typical physical hallmark of tumors. Mounting evidence indicates that elevated solid stress drives metastasis and affects prognosis. However, the molecular mechanism of how cancer cells sense solid stress, thereby exacerbating malignancy, remains elusive. In this study, our clinical data suggest that elevated stress in metastatic solid tumors is highly associated with the expression of cytoskeleton-associated protein 4 (CKAP4). Intriguingly, CKAP4, as a sensitive intracellular mechanosensor, responds specifically to solid stress in a subset of studied tumor micro-environmental elements through liquid-liquid phase separation. These micron-scaled CKAP4 puncta adhere tightly onto microtubules and dramatically reorchestrate their curvature and branching to enhance cell spreading, which, as a result, boosts cancer cell motility and facilitates distant metastasis in vivo. Mechanistically, the intrinsically disordered region 1 (IDR1) of CKAP4 binds to microtubules, while IDR2 governs phase separation due to the Cav1.2-dependent calcium influx, which collectively remodels microtubules. These findings reveal an unprecedented mechanism of how cancer cells sense solid stress for cancer malignancy and bridge the gap between cancer physics and cancer cell biology.

EPCO-15. INTRA-TUMORAL HETEROGENEITY OF DNA METHYLATION PROFILING AND CELLULAR COMPOSITION IN GLIOBLASTOMA

Abstract BACKGROUND DNA methylation profiling has become an important diagnostic and exploratory tool in neuro-oncology. Intra-tumoral molecular heterogeneity is a crucial factor in the treatment resistance of IDH-wildtype glioblastoma (GBM). However, the intra-tumoral heterogeneity of methylation profiles in GBM remains poorly elucidated. This study aimed to investigate the intra-tumoral heterogeneity of DNA methylation subclasses and deconvolve methylation profiles to infer the cellular composition of the tumor. METHODS We conducted genome-wide DNA methylation analysis and applied the DKFZ/Heidelberg CNS tumor classifier to FFPE tissue samples from a multi-sampling cohort with multiple samples per patient and a single-sampling cohort with one sample per patient. Only samples classified as GBM were included. We conducted two different methylation-based deconvolution analyses. Deconvolution 1 inferred the fractions of tumor subtypes (RTK_I, RTK_II, MES_TYP, and MES_ATYP) and cell types in the microenvironment. Deconvolution 2 estimated the abundance of malignant cell states (stem-like and differentiated cell components) and cell types in the microenvironment. RESULTS The multi-sampling cohort included 32 patients with 113 samples, and the single-sampling cohort included 87 patients with 87 samples. In the multi-sampling cohort, 8 patients (25%) exhibited intra-patient heterogeneity in methylation subclasses across their samples. Deconvolution 1 indicated that each tumor subclass component was heterogeneously distributed within samples, and the abundance of the myeloid cell component correlated with the MES_TYP component across samples. Deconvolution 2 similarly revealed heterogeneity in the immune component abundance across samples within patients. Integrating the results of Deconvolution 1 and Deconvolution 2 using the entire cohort revealed significant correlations between RTK_I and stem-like, RTK_II and differentiated, and MES_TYP and immune components. These findings were also replicated in TCGA dataset. CONCLUSIONS Our findings underscore the importance of considering intra-tumoral heterogeneity in methylation profiles and the biological characteristics of each methylation subclass in developing novel GBM therapies.

Moisturizing and antioxidant factors of skin barrier restoring cream with shea butter, silkflo and vitamin E in human keratinocyte cells.

Moisturizers are integral to daily skincare routines, reflecting the increasing trend among people towards cosmetic products, particularly for skin care. They significantly contribute to preserving skin health, particularly by regulating the epidermal barrier and moisture levels within the skin. This study aims to explore the moisturizing and antioxidant effect of skin barrier restoring cream Moiz MM (MZ) with shea butter, silkflo and vitamin E by investigating its protective effect against oxidative stress-induced cellular damage and therapeutic mechanisms in human keratinocytes cells (HaCaT). The invitro antioxidant activity of MZ was evaluated by DPPH, ABTS and NO assays. For the cell culture study, HaCaT cells were cultured and stimulated using H2O2 and then treated with different concentrations of MZ. Then, it was subjected to DCFH-DA staining, reverse transcriptase PCR and western blot analysis for the evaluation of various skin-moisture-related components in human keratinocyte cells. Type I procollagen was examined using ELISA technique. The results highlighted that oxidative stress in HaCaT cells decreased type I procollagen synthesis, while MZ treatment significantly increased the synthesis. Moreover, the viability of HaCaT cells was not affected in the presence of MZ, which demonstrates its non-toxic effect. Furthermore, MZ can counteract H2O2-mediated oxidative stress by enhancing the antioxidant enzyme activity such as superoxide dismutase and catalase, and decrease reactive oxygen species generation in skin cells. Additionally, MZ greatly promotes hyaluronic acid production by enhancing the expression of the hyaluronic acid synthase-1 gene and Aquaporin 3 protein. This study suggests that MZ has the potential to serve as a moisturizing and antioxidant skincare formula.

NIMG-27. RADIOMETHYLOMICS FOR NON-INVASIVE PREDICTION OF TUMOR PHENOTYPES AND SURVIVAL IN GLIOBLASTOMA

Abstract BACKGROUND DNA methylation profiling has become a pivotal tool in neuro-oncology. However, little is known about the impact of methylation profiling on radiological imaging in IDH-wildtype glioblastoma (GBM). This study explored the potential of radiomics to non-invasively predict molecular characteristics based on the methylation profiling of GBM. METHODS This study included a multi-sampling cohort of 32 patients with 113 samples and a single-sampling cohort of 87 patients with 87 samples, which underwent genome-wide methylation analysis and were classified as GBM by the DKFZ/Heidelberg CNS tumor classifier. We performed two different methylation-based deconvolution analyses. Deconvolution 1 estimated the fractions of tumor subtypes (RTK_I, RTK_II, MES_TYP, and MES_ATYP) and cell types in the microenvironment. Deconvolution 2 inferred the abundance of malignant cell states (stem-like and differentiated cell components) and cell types in the microenvironment. We derived 6084 radiomic features from preoperative multi-parametric MRI in each patient. RESULTS In the multi-sampling cohort, Deconvolution 1 and Deconvolution 2 demonstrated that tumor subclass components and immune components exhibited heterogeneous distribution across samples within patients; however, the stem-like to differentiated cell ratio was preserved across samples within patients. In the entire cohort, patients with tumors exhibiting a high proportion of the stem-like component, defined as stem-like tumors, had significantly shorter overall survival. Stem-like tumors exhibited significantly higher levels of the RTK_I component and lower proportions of the RTK_II component compared to non-stem-like tumors. A machine learning model that utilizes support vector machines to classify stem-like tumors using radiomic features achieved a mean AUC of 0.71 (95% confidence interval: 0.56–0.85) in nested cross-validation. CONCLUSIONS We showed that radiomethylomics can be used to predict tumor composition and patient survival from preoperative MRI in GBM. Based on these promising results we will increase our power with additional samples to facilitate a personalized approach to GBM management.

Cell components of tumor microenvironment in lung adenocarcinoma: Promising targets for small-molecule compounds.

Lung cancer is one of the most lethal tumors in the world with a 5-year overall survival rate of less than 20%, mainly including lung adenocarcinoma (LUAD). Tumor microenvironment (TME) has become a new research focus in the treatment of lung cancer. The TME is heterogeneous in composition and consists of cellular components, growth factors, proteases, and extracellular matrix. The various cellular components exert a different role in apoptosis, metastasis, or proliferation of lung cancer cells through different pathways, thus contributing to the treatment of adenocarcinoma and potentially facilitating novel therapeutic methods. This review summarizes the research progress on different cellular components with cell-cell interactions in the TME of LUAD, along with their corresponding drug candidates, suggesting that targeting cellular components in the TME of LUAD holds great promise for future theraputic development.

Unveiling the distinctive variations in multi-omics triggered by TP53 mutation in lung cancer subtypes: An insight from interaction among intratumoral microbiota, tumor microenvironment, and pathology

The TP53 mutation is one of the most common gene mutations in non-small cell lung cancer (NSCLC) and plays a significant role in the occurrence, development, and prognosis of both lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). Recent studies have also suggested the predictive value of TP53 mutations in the response to immunotherapy for NSCLC. It is known that intratumoral microbiota, tumor immune microenvironment (TIME) and histology are associated with the roles of TP53 mutation in NSCLC. However, the intrinsic associations among these three factors and their underlying interaction with TP53 mutation are not well understood. Additionally, the potential of predicting TP53 mutations using deep learning methods has not yet been fully evaluated. In this paper, we comprehensively evaluated the tissue microbiome, host gene expression characteristics, and histopathological slides of 992 NSCLC patients obtained from the cancer genome atlas (TCGA) and validated the findings using multi-omics data of 332 NSCLC patients from the Clinical Proteomic Tumor Analysis Consortium (CPTAC). Compared to LUSC, LUAD exhibited more substantial differences between patients with and without TP53 mutation in all three aspects. In LUAD, our results imply underlying links between the tissue microbiome and immune cell components in the TIME, and show that the abundance of immune cells is reflected in histology slides. Furthermore, we propose a novel multimodal deep learning model that focuses on histopathology images, which achieves an area under the curve (AUC) of 0.84 in LUAD. In summary, TP53 mutation of LUAD resulted more significant changes in intratumoral microbiota, TIME and histology than that of LUSC. And histopathology images can be used to predict TP53 mutation in LUAD with reasonable accuracy.

A Comprehensive Approach to Optimization of Silicon-Based Solar Cells

In this work, we report a detailed scheme of computational optimization of solar cell structures and parameters using PC1D and AFORS-HET codes. Each parameter’s influence on the properties of the components of heterojunction silicon-based solar cells (HIT) has been thoroughly examined. The proposed approach follows a stringent sequence of steps to optimize various parameters of the studied HITs. Furthermore, we have revealed the effects of the metal-semiconductor contact, and a model of a photocell with an ohmic contact and a Schottky contact has been simulated. The optimal model of HIT for available materials has been proposed and fabricated based on the results of these simulations. A comparison of predicted and measured performance unequivocally demonstrates the efficiency of the proposed scheme in developing silicon-based HITs, providing reassurance about its practical application.

Cancer immunogenic cell death via pyroptosis with CXCR4-targeted nanotoxins in hepatocellular carcinoma

IntroductionCytotoxic agents have shown limited benefits in hepatocellular carcinoma (HCC), mediated in part by the lack of targeting. As cell-penetrating peptides (CPPs) are capable of delivering various biologically active molecules into cells, including protein, peptides, small chemo-drugs, and nucleic acid with or without targeting, we developed T22-PE24, a CXCR4-targeted self-assembling cytotoxic nanotoxin, to effectively induce HCC pyroptosis.MethodsT22 incorporating enhanced green fluorescent protein (EGFP) or PE24 was purified from DE3 bacterial cells and characterized using transmission electron microscopy, the Zetasizer Nano®, and SEC-HPLC. The internalization effect of T22-EGFP was detected by flow cytometry system (FCS) in CXCR4+/LM3(CXCR4−) HCC cells. The CCK8, lactate dehydrogenase (LDH) release, Western blot, and nude mice HCC models were used to estimate the cell viability of T22-PE24. The complete-immunity HCC tumor-bearing mice model was used to assess the immune response of T22-PE24.ResultsThe round shape under transmission electron microscopy, 49.4 nm hydrodynamic diameter, and −33.33 mV zeta potential indicated that T22-PE24 self-assembled into nanoparticles. T22 incorporating EGFP selectively internalized in CXCR4+ HCC cells and showed no accumulation in CXCR4-knockout HCC cells. The T22-PE24 nanotoxin induced HCC pyroptosis via the caspase-3/GSDME signaling pathway and suppressed tumor growth in the absence of histological alterations in normal organs. Using the complete-immunity HCC tumor-bearing mice model, we found that T22-PE24 nanotoxin effectively induces the global reprogramming of cell components of the immune tumor microenvironment, leading to enhanced antitumor effects compared to those observed in immunodeficient mice.ConclusionOur findings demonstrate the activation of the innate immune response in HCC by inducing pyroptosis with T22-PE24 nanotoxin treatment and support an implementation of this strategy for HCC treatment.

Blood-brain barrier permeability increases with the differentiation of glioblastoma cells in vitro

BackgroundGlioblastoma multiforme (GBM) is an aggressive tumor, difficult to treat pharmacologically because of the blood-brain barrier (BBB), which is rich in ATP-binding cassette (ABC) transporters and tight junction (TJ) proteins. The BBB is disrupted within GBM bulk, but it is competent in brain-adjacent-to-tumor areas, where eventual GBM foci can trigger tumor relapse. How GBM cells influence the permeability of BBB is poorly investigated.MethodsTo clarify this point, we co-cultured human BBB models with 3 patient-derived GBM cells, after separating from each tumor the stem cell/neurosphere (SC/NS) and the differentiated/adherent cell (AC) components. Also, we set up cultures of BBB cells with the conditioned medium of NS or AC, enriched or depleted of IL-6. Extracellular cytokines were measured by protein arrays and ELISA. The intracellular signaling in BBB cells was measured by immunoblotting, in the presence of STAT3 pharmacological inhibitor or specific PROTAC. The competence of BBB was evaluated by permeability assays and TEER measurement.ResultsThe presence of GBM cells or their conditioned medium increased the permeability to doxorubicin, mitoxantrone and dextran-70, decreased TEER, down-regulated ABC transporters and TJ proteins at the transcriptional level. These effects were higher with AC or their medium than with NS. The secretome analysis identified IL-6 as significantly more produced by AC than by NS. Notably, AC-conditioned medium treated with an IL-6 neutralizing antibody reduced the BBB permeability to NS levels, while NS-conditioned medium enriched with IL-6 increased BBB permeability to AC levels. Mechanistically, IL-6 released by AC GBM cells activated STAT3 in BBB cells. In turn, STAT3 down-regulated ABC transporter and TJ expression, increased permeability and decreased TEER. The same effects were obtained in BBB cells treated with STA-21, a pharmacological inhibitor of STAT3, or with a PROTAC targeting STAT3.ConclusionsOur work demonstrates for the first time that the degree of GBM differentiation influences BBB permeability. The crosstalk between GBM cells that release IL-6 and BBB cells that respond by activating STAT3, controls the expression of ABC transporters and TJ proteins on BBB. These results may pave the way for novel therapeutic tools to tune BBB permeability and improve drug delivery to GBM.

Immunomodulatory Potential of Melatonin in Immunosuppression: An in-vivo Study.

To assess the immunomodulatory effects of melatonin on the acquired immunity of immunosuppressed male Wistar rats by checking Interleukin 6 (IL-6) levels, total leucocyte counts (TLC), and differential leucocyte counts. Experimental study. Place and Duration of the Study: Animal laboratory, CMH Lahore Medical College, from June to October 2023. Fifty male Wistar rats, weighing 180g to 200g, were included in this study with 10 rats in each of the five groups namely cyclophosphamide (CP), CP + melatonin, CP + immunomodulator, melatonin-only, and control. A calculated dose of CP was administered intraperitoneally for 30 days (from 13/06/23 to 13/07/23) to each group except the control groups. After this, the experimental group was given melatonin CP + Melatonin for 7 days (from 14/07/23 to 20/07/23). CP + Immunomodulatory group was kept for comparison of immunomodulatory effects. Blood samples were drawn from all 5 groups. IL-6 was estimated through ELISA. Other parameters assessed were TLC and absolute differential leucocyte counts. Melatonin increased IL-6 levels significantly (p = 0.042) as well as the TLC levels (p <0.001) compared to the immuno-suppressed CP group. Melatonin was seen to have an upregulation of IL-6 levels in immunosuppression compared to the administration of immunomodulator preparation (p = 0.506) which was not as effective. Administration of melatonin significantly increased the TLC, neutrophil, lymphocyte, monocyte, and eosinophil count compared to known immunomodulators. Melatonin as a supplement may have some role in activating multiple immune response processes in immune-depressed states. It was also established that it allows quick recovery of cell components from immunosuppressed states. Melatonin, Immunomodulation, Rats, Interleukin-6, Acquired immunity, Cyclophosphamide.

Understanding the spleen response of Russian sturgeon (Acipenser gueldenstaedtii) dealing with chronic heat stress and Aeromonas hydrophila challenge

Sturgeon aquaculture has grown in recent years, driven by increasing global demand for its highly valued products. Russian sturgeon (Acipenser gueldenstaedtii), recognised as one of the most valuable species for caviar production, is farmed in several warm-temperate regions. However, the substantial temperature increase due to global warming represents a challenge for developing sturgeon aquaculture. Previously we demonstrated that Russian sturgeon under chronic heat stress (CHS) exhibited a liver metabolic reprogramming to meet energy demands, weakening their innate defences and leading to increased mortality and economic losses. Here, we used RNA-seq technology to analyse regulated genes in the spleen of Russian sturgeons exposed to CHS and challenged with Aeromonas hydrophila. The assembly gave 253,415 unigenes, with 13.7 % having at least one reliable functional annotation. We found that CHS caused mild splenitis and upregulated genes related to protein folding, heat shock response, apoptosis and autophagy while downregulated genes associated with the cell cycle. The cell cycle arrest was maintained upon A. hydrophila challenge in heat-stressed fish, potentially inducing cell senescence. Surprisingly, immunoglobulin heavy and light chains were upregulated in the spleen of stressed sturgeons but not in those maintained at tolerable temperatures; however, no changes in IgM serum levels were observed in any condition. Our findings indicate that long-term exposure to non-tolerable temperatures induced a heat shock response and activated apoptosis and autophagy processes in the spleen. These mechanisms may enable the control of tissue damage and facilitate the recycling of cell components in a condition where the nutrient supply by the liver might be insufficient. Stressed sturgeons challenged with A. hydrophila maintain these mechanisms, which could culminate in cellular senescence.

Immunomodulatory Potential of Melatonin in Immunosuppression: An in-vivo Study.

To assess the immunomodulatory effects of melatonin on the acquired immunity of immunosuppressed male Wistar rats by checking Interleukin 6 (IL-6) levels, total leucocyte counts (TLC), and differential leucocyte counts. Experimental study. Place and Duration of the Study: Animal laboratory, CMH Lahore Medical College, from June to October 2023. Fifty male Wistar rats, weighing 180g to 200g, were included in this study with 10 rats in each of the five groups namely cyclophosphamide (CP), CP + melatonin, CP + immunomodulator, melatonin-only, and control. A calculated dose of CP was administered intraperitoneally for 30 days (from 13/06/23 to 13/07/23) to each group except the control groups. After this, the experimental group was given melatonin CP + Melatonin for 7 days (from 14/07/23 to 20/07/23). CP + Immunomodulatory group was kept for comparison of immunomodulatory effects. Blood samples were drawn from all 5 groups. IL-6 was estimated through ELISA. Other parameters assessed were TLC and absolute differential leucocyte counts. Melatonin increased IL-6 levels significantly (p = 0.042) as well as the TLC levels (p <0.001) compared to the immuno-suppressed CP group. Melatonin was seen to have an upregulation of IL-6 levels in immunosuppression compared to the administration of immunomodulator preparation (p = 0.506) which was not as effective. Administration of melatonin significantly increased the TLC, neutrophil, lymphocyte, monocyte, and eosinophil count compared to known immunomodulators. Melatonin as a supplement may have some role in activating multiple immune response processes in immune-depressed states. It was also established that it allows quick recovery of cell components from immunosuppressed states. Melatonin, Immunomodulation, Rats, Interleukin-6, Acquired immunity, Cyclophosphamide.

Spatial Architecture of Single-cell and Vasculature in Tumor Microenvironment Predicts Clinical Outcomes in Triple-Negative Breast Cancer

Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer with limited treatment options, which warrants the identification of novel therapeutic targets. Deciphering nuances in the tumor microenvironment (TME) may unveil insightful links between anti-tumor immunity and clinical outcomes, yet such connections remain underexplored. Here we employed a dataset derived from imaging mass cytometry of 71 TNBC patient specimens at single-cell resolution and performed in-depth quantifications with a suite of multi-scale computational algorithms. The TNBC TME reflected a heterogeneous ecosystem with high spatial and compositional heterogeneity. Spatial analysis identified ten recurrent cellular neighborhoods (CNs) – a collection of local TME characteristics with unique cell components. The prevalence of CNs enriched with B cells, fibroblasts, and tumor cells, in conjunction with vascular density and perivasculature immune profiles, could significantly enrich for long-term survivors. Furthermore, relative spatial colocalization of SMAhi fibroblasts and tumor cells compared to B cells correlated significantly with favorable clinical outcomes. Using a deep learning model trained on engineered spatial data, we can predict with high accuracy (mean AUC of 5-fold cross-validation = 0.71) how a separate cohort of patients in the NeoTRIP clinical trial will respond to treatment based on baseline TME features. These data reinforce that the TME architecture is structured in cellular compositions, spatial organizations, vasculature biology, and molecular profiles, and suggest novel imaging-based biomarkers for treatment development in the context of TNBC.