Unraveling the Complexity and Advancements of Transdifferentiation Technologies in the Biomedical Field and Their Potential Clinical Relevance
Purusottam Mishra · Izabella Biesiada · Payal Gupta · Saeid Ghavami · Jarosław Markowski · Marek J. Łos
Abstract.
Chronic diseases such as cancer, autoimmunity, and organ failure currently depend on conventional pharmaceutical treatment, which may cause detrimental side effects in the long term. In this regard, cell-based therapy has emerged as a suitable alternative for treating these chronic diseases. Transdifferentiation technologies have evolved as a suitable therapeutic alternative that converts one differentiated somatic cell into another phenotype by using transcription factors (TFs), small molecules, or small, single-stranded, non-coding RNA molecules (miRNA). The transdifferentiation techniques rely on simple, fast, standardized, and versatile protocols with minimal chance of tumorigenicity and genotoxicity. However, there are still challenges and limitations that need to be addressed to enhance their clinical translation percentage in the near future. Taking this into account, we have delineated the features and strategies used in the transdifferentiation techniques. Then, we delved into different intermediate states that were attained during transdifferentiation. Advancements in transdifferentiation techniques in the field of tissue engineering, autoimmunity, and cancer therapy were dissected. Furthermore, limitations, challenges, and future perspectives are outlined in this review to provide a whole new picture of the transdifferentiation techniques. Advancements in molecular biology, interdisciplinary research, bioinformatics, and artificial intelligence will push the frontiers of this technology further to establish new avenues for biomedical research.
Keywords: Cell reprogramming • Osteoblasts • Transdifferentiation • Stem cells • Tissue engineering • Transcription factors
Quantification of Citrullinated Histone H3 as a Marker for Neutrophil Extracellular Traps Correlated to Clinical Characteristics of Patients with Systemic Lupus Erythematosus
Maciej Dubaj · Iwona Homa-Mlak · Aleksandra Majdan · Radosław Mlak · Maria Majdan · Teresa Małecka-Massalska
Abstract.
Systemic lupus erythematosus (SLE) is an autoimmune disease whose pathogenesis is not fully understood to date. One of the suggested mechanisms for its development is NETosis, which involves the release of a specific network consisting of chromatin, proteins, and enzymes from neutrophils, stimulating the immune system. One of its markers is citrullinated histone H3 (H3Cit). This study aimed to evaluate the correlation of H3Cit levels with the clinical characteristics of 80 LE patients. Levels of H3Cit in the subjects’ serum were quantified spectrophotometrically. Statistical analysis was performed using MedCalc 15.8 and Statistica 13.3. Significantly higher H3Cit levels were found in patients with arthralgia (medians [interquartile range] [IQR]: 1.67 [1.67–1.69] vs. 1.67 [1.62–1.68], p = 0.0150, respectively) and reduced complement component C4 levels compared to patients without these conditions (medians [IQR]: 1.68 [1.67–1.70] vs. 1.68 [1.67–1.69], p = 0.0297, respectively). A significant weak negative correlation was observed between H3Cit levels and leukocytosis (rho = −0.2602, p = 0.0309) and reduced complement component C3 levels (rho = −0.2442, p = 0.0447) and a weak positive correlation with anti-double stranded DNA (anti-dsDNA) antibody levels (rho = 0.3794, p = 0.0036). Moreover, the clinical utility of the H3Cit assay in differentiating patients with arthralgia (area under the curve [AUC] = 0.709, p = 0.0115), seizures (AUC = 0.813, p = 0.0005), hepatomegaly (AUC = 0.746, p = 0.0111), and reduced levels of complement component C4 (AUC = 0.662, p = 0.0224) and without the above conditions was noted.
Keywords: Autoimmune diseases • Systemic lupus erythematosus • Histone H3 • Citrullination • Extracellular Traps
CRISPR/Cas Systems as Diagnostic and Potential Therapeutic Tools for Enterohemorrhagic Escherichia coli
Agnieszka Bogut · Anna Kołodziejek · Scott A. Minnich · Carolyn J. Hovde
Abstract.
Following its discovery as an adaptive immune system in prokaryotes, the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated proteins (Cas) system has been developed into a multifaceted genome editing tool. This review compiles findings aimed at implementation of this technology for selective elimination or attenuation of enterohemorrhagic Escherichia coli (EHEC). EHEC are important zoonotic foodborne pathogens that cause hemorrhagic colitis and can progress to the life-threatening hemolytic uremic syndrome (HUS). Advancements in the application of CRISPR methodology include laboratory detection and identification of EHEC, genotyping, screening for pathogenic potential, and engineering probiotics to reduce microbial shedding by cattle, the primary source of human infection. Genetically engineered phages or conjugative plasmids have been designed to target and inactivate genes whose products are critical for EHEC virulence.
Keywords: EHEC • CRISPR/Cas • Cas9 • Cas12 • Probiotics • O157:H7
Antinuclear Antibodies in Non-Rheumatic Diseases
Nikita Niranjan Kumar · Samir Ahmad Dit Al Hakim · Bogna Grygiel-Górniak
Abstract.
Antinuclear antibodies (ANAs) are critical immunological markers commonly associated with various connective tissue diseases (CTDs). However, these autoantibodies are also detectable in healthy individuals, patients with non-rheumatic autoimmune diseases, those with viral infections, and subjects using specific medications (such as procainamide, hydralazine, and minocycline) that can lead to drug-induced ANA elevation. The standard method for ANA detection is indirect immunofluorescence, a process that requires precision and thoroughness as it assesses both titer and fluorescence patterns. Additionally, immunoblotting and enzyme-linked immunosorbent assay (ELISA) are recommended to identify specific ANAs precisely, highlighting the importance of precision in ANA detection. This review explores the advantages and limitations of current ANA detection methods. It also describes the clinical implications of ANA presence in non-rheumatic diseases, including autoimmune disorders, infectious conditions, non-autoimmune and non-infectious diseases, and autoimmune cutaneous diseases. The presence of elevated ANA titers in these contexts can complicate clinical decision-making, as the diagnostic value of ANA testing alone is limited in non-rheumatic conditions. However, despite these limitations, ANA remains a key component in diagnosing and prognosis systemic CTDs, as it can indicate isease activity, severity, and response to treatment, which is of utmost importance in rheumatology and internal medicine. This paper provides a comprehensive review of the role of ANA in non-rheumatic diseases. It focuses on ANA diagnostic and prognostic significance and offers valuable insights for clinical practice.
Keywords: Antinuclear antibodies • Methods of detection • Autoimmune non-rheumatic diseases • Infectious diseases
Growing Challenges of Lung Infections with Non-tuberculous Mycobacteria in Immunocompromised Patients: Epidemiology and Treatment
Weronika Burzyńska · Marek Fol · Magdalena Druszczynska
Abstract.
Non-tuberculous mycobacteria (NTM) are increasingly recognized as opportunistic pathogens in humans and animals, particularly affecting those with compromised immune systems. These bacteria encompass a diverse group of mycobacterial species that are responsible for a range of infections, with pulmonary and skin-related conditions being the most common. The rise in NTM infections in recent years is a growing concern for healthcare, highlighting the urgent need to improve our understanding of NTM epidemiology and treatment strategies. This article reviews the NTM species associated with lung infections in immunocompromised patients and underscores the critical importance of advancing diagnostic and therapeutic approaches. The review is based on a thorough analysis of scientific literature from databases such as PubMed, Scopus, and ScienceDirect, covering studies up to June 2024. Through this comprehensive analysis, the article aims to provide detailed insights into the complexities of NTM diseases and spur further research and innovation in combating these challenging infections.
Keywords: Non-tuberculous mycobacteria • Lung infections • Immunodeficiency • atypical mycobacteria
Isorhynchophylline Inhibits NLRP3 Inflammasome and Improves
Gestational Diabetes
Li Pu · Li Chen · Kun Yu · Yueming Zhang · Caifeng Wang · Feizhou Jiang · Jia Shi · Jingjing Meng
Abstract.
This study aims to investigate the effects and underlying mechanisms of isorhynchophylline (IRN) on gestational diabetes mellitus (GDM). The db/+ mice were randomly divided into four groups: GDM, GDM + IRN (20 mg/kg), and GDM + IRN (40 mg/kg). Blood glucose and insulin tolerance were assessed using intraperitoneal glucose tolerance tests (IPGTTs) and intraperitoneal insulin tolerance tests (IPITTs) on gestational day 10. On gestational day 20, placental inflammation (tumor necrosis factor [TNF]-α, interleukin [IL]-6, IL-1β), oxidative stress markers (malondialdehyde [MDA], SOD, glutathione peroxidase [GPx], and glutathione [GSH]), and nuclear factor-κB/NOD-like receptor protein 3 [NLRP3] inflammasome activity were measured using enzyme-linked immunosorbent assay (ELISA), immunoblotting, and biochemical assays. IRN significantly improved blood glucose levels and insulin tolerance in GDM mice. IRN treatment reduced placental inflammation. In addition, oxidative stress in the placenta was alleviated in the IRN-treated groups, leading to improved placental function and healthier fetal development. The birth weight of offspring was higher in the IRN-treated groups compared with untreated GDM mice. Furthermore, IRN inhibited the activation of the NLRP3 pathway. IRN significantly improves metabolic and inflammatory parameters in GDM through the NF-κB/NLRP3 pathway, highlighting its potential therapeutic benefits for managing GDM and improving maternal and fetal outcomes.
Keywords: Gestational diabetes mellitus • Isorhynchophylline • Insulin resistance • Inflammation • NF-κB/NLRP3 pathway
Isorhynchophylline Inhibits NLRP3 Inflammasome and Improves
Gestational Diabetes
Li Pu · Li Chen · Kun Yu · Yueming Zhang · Caifeng Wang · Feizhou Jiang · Jia Shi · Jingjing Meng
Abstract.
This study aims to investigate the effects and underlying mechanisms of isorhynchophylline (IRN) on gestational diabetes mellitus (GDM). The db/+ mice were randomly divided into four groups: GDM, GDM + IRN (20 mg/kg), and GDM + IRN (40 mg/kg). Blood glucose and insulin tolerance were assessed using intraperitoneal glucose tolerance tests (IPGTTs) and intraperitoneal insulin tolerance tests (IPITTs) on gestational day 10. On gestational day 20, placental inflammation (tumor necrosis factor [TNF]-α, interleukin [IL]-6, IL-1β), oxidative stress markers (malondialdehyde [MDA], SOD, glutathione peroxidase [GPx], and glutathione [GSH]), and nuclear factor-κB/NOD-like receptor protein 3 [NLRP3] inflammasome activity were measured using enzyme-linked immunosorbent assay (ELISA), immunoblotting, and biochemical assays. IRN significantly improved blood glucose levels and insulin tolerance in GDM mice. IRN treatment reduced placental inflammation. In addition, oxidative stress in the placenta was alleviated in the IRN-treated groups, leading to improved placental function and healthier fetal development. The birth weight of offspring was higher in the IRN-treated groups compared with untreated GDM mice. Furthermore, IRN inhibited the activation of the NLRP3 pathway. IRN significantly improves metabolic and inflammatory parameters in GDM through the NF-κB/NLRP3 pathway, highlighting its potential therapeutic benefits for managing GDM and improving maternal and fetal outcomes.
Keywords: Gestational diabetes mellitus • Isorhynchophylline • Insulin resistance • Inflammation • NF-κB/NLRP3 pathway
Protective Effect of the Human Epineural Patch Application after Sciatic Nerve Crush Injury Followed by Nerve Transection and End-to-End Repair
Maria Siemionow · Weronika Radecka · Katarzyna Kozlowska · Lucile Chambily · Sonia Brodowska · Dominika Kuc · Gabrielle Filipek · Katarzyna Budzynska
Abstract.
Nerve regeneration under unfavorable wound conditions remains challenging. We introduce the human epineural patch (hEP) as a novel nerve protector for posttrauma applications, comparing its regenerative efficacy with that of the human amniotic membrane (hAM). Following crush injury, transection, and end-to-end repair (CTR), 36 athymic nude rats were randomly assigned to six experimental groups (n = 6 each): control (no-protection), hEP, or hAM application post-repair. Assessments at 6 weeks and 12 weeks included functional evaluation (Toe-Spread and Pinprick tests), gastrocnemius muscle index (GMI), histomorphometric analysis (myelin thickness, axonal density, fiber diameter, percentage of myelinated fibers), and immunofluorescence staining for neurogenic, angiogenic, and immunogenic markers. The hEP group exhibited superior motor (3.167 ± 0.167) and sensory (3.500 ± 0.212) recovery and GMI values (0.955 ± 0.014), compared with the No protection group (p < 0.05). Myelin thickness (3.480 ± 0.019 μm, p < 0.0001), fiber diameter (10.788 ± 0.197 μm, p < 0.05), and myelinated fiber percentage (89.841% ± 0.453%, p < 0.01) were significantly higher in the hEP group. At 12 weeks, hEP application significantly increased the expression of Laminin B (2.083 ± 0.083), nerve growth factor (NGF) (1.750 ± 0.250), and vascular endothelial growth factor (VEGF) (2.667 ± 0.333), corresponding with improved function. The application of hEP at the sciatic nerve repair site after CTR injury significantly enhanced nerve regeneration compared with hAM. This study introduces hEP as a promising alternative nerve protector for traumatic nerve injuries.
Keywords: Peripheral nerve repair • Human epineural patch • Human amniotic membrane • Allografts • Nerve regeneration • Regenerative medicine
Sanguinarine Inhibits Cell Growth in EBV-Positive Diffuse Large B-Cell Lymphoma
Suli Lu · Dae-jung Yang
Abstract.
To investigate the effects of Sanguinarine (SAG) on the progression of diffuse large B-cell lymphoma (DLBCL) and to explore its underlying mechanism, this study utilized Epstein–Barr virus (EBV)-positive DLBCL cell lines, FARAGE, and GM12878S. Cell counting kit-8 and bromodeoxyuridine assays were used to assess the effects of SAG on the cell proliferation. Flow cytometry and immunoblotting were employed to analyze cell cycle arrest and apoptosis. Additionally, the molecular mechanism was explored through further immunoblotting analysis of the mechanism. SAG suppressed the growth of EBV-positive DLBCL cells. Furthermore, SAG induced cell cycle arrest and promoted apoptosis in these cells. Mechanistically, SAG suppressed the Wnt/β-catenin pathway, thereby suppressing DLBCL progression in vitro. SAG effectively inhibits cell growth and induces apoptosis in EBV-positive DLBCL via Wnt/β-catenin pathway, offering potential therapeutic insights for this lymphoma subtype.
Keywords: Sanguinarine • EBV-positive DLBCL • Cell cycle • Apoptosis • Wnt/β-catenin
Thrombotic Markers in Plasma as Predictors of Response in Rheumatoid Arthritis Patients Treated with Baricitinib – Pilot Observation
Anna Felis-Giemza, · Kornelia Chmurzynska · Jakub Wronski · Paulina Klimek · Anna Kornatka · Wlodzimierz Maslinski · Marzena Ciechomska · Magdalena Massalska
Abstract.
Both disease and treatment carry the risk of thrombotic events in patients with rheumatoid arthritis (RA). This pilot study aimed to assess changes in thrombotic markers in plasma and their potential role as predictors of response during early baricitinib treatment. The concentrations of antithrombin III (ATIII) activity, D-dimer (DD), fibrinogen, and homocysteine (HCY) were evaluated in RA subjects before and 3 months after the treatment. At baseline, the RA group had higher DD (1472.3 ± 349.2) and fibrinogen (410.4 ± 29.5) compared with healthy controls (HC; 450.3 ± 54.5; p = 0.0002 and 334.9 ± 19.2; p = 0.04, respectively). with no differences in ATIII and HCY. After 3 months, we observed a significant increase in HCY (10.7 ± 0.6 vs. 9.1 ± 0.5; p = 0.018) and ATIII (119.7 ± 2.7 vs. 110.4 ± 3.2; p = 0.004), the latter correlated negatively with disease activity score 28 (DAS28; r = –0686, p < 0.002). After 3 months of baricitinib therapy, the patients were divided into moderate responders (MR) and good responders (GR) groups according to EULAR criteria. At baseline, MR had higher DD (1639.2 ± 550.5 vs. 450.3 ± 54.5; p < 0.0001) and lower ATIII (105.3 ± 3.6 vs. 115.1 ± 2.7; p = 0.043) compared with HC. Thrombotic parameters in the first 3 months of baricitinib treatment were mostly in line with current findings concerning the RA population. Increased levels of DD together with low ATIII concentrations seem to predispose to a moderate response to baricitinib treatment.
Keywords: Antithrombin III • Cardiometabolic risk biomarkers • Fibrinogen • Homocystein • Rheumatoid arthritis • Baricitinib
Exploring Diagnostic Markers and Therapeutic Targets in Parkinson’s Disease: A Comprehensive 1H-NMR Metabolomic Analysis – Systematic Review
Andrzej Wasilewski · Eliza Wasilewska · Agata Serrafi
Abstract.
Parkinson’s disease (PD) affects millions of people globally. Accurate early diagnosis remains a challenge due to the lack of specific biomarkers. This systematic review explores the potential of 1H-NMR metabolomics in identifying diagnostic markers and therapeutic targets for PD. A comprehensive analysis was conducted across databases such as Scopus, Web of Science, PubMed, and Embase, focusing on studies that utilized 1H-NMR spectroscopy to profile metabolites associated with PD progression. The review identifies key metabolites—glutamate, taurine, myo-inositol, glutamine, and creatine—that play critical roles in the pathophysiology of PD. Glutamate, linked to excitotoxicity and neuronal degeneration, emerges as a prominent target for therapeutic intervention, while taurine is associated with oxidative stress. Myo-inositol, a key regulator of autophagy, underscores the biochemical dysregulation associated with PD, similar to glutamine and glutamate. Creatine’s role in neuronal energy metabolism suggests potential avenues for treatment focused on energy supplementation. The reproducibility of metabolite findings varied, indicating the complexity of PD’s metabolomic landscape. Despite challenges in consistency, these metabolites hold promise as biomarkers for diagnosing PD and tracking disease progression. The review underscores the need for further validation of these markers and their integration with other omics technologies to enhance PD management. By identifying key metabolic pathways, this study opens new directions for personalized medicine, offering potential therapeutic targets to slow disease progression and improve patient
outcomes.
Keywords: 1H-NMR spectroscopy • Parkinson’s disease • Therapeutic targets • Oxidative stress • Metabolite profiling • Metabolomic
Intraarterial Infusion of Lidocaine is Superior to the Subcutaneous Injection of Low Molecular Weight Heparin for Improving the Course of Cerulein-Induced Acute Pancreatitis in Rats
Ryszard Antkowiak · Lukasz Antkowiak · Zbigniew Arent · Bogna Drozdzowska · Anna Kasperczuk · Jacek Bialecki · Agnieszka Pietsch-Fulbiszewska · Pawel Domoslawski · Agata Cieslik-Bielecka · Marek Kucharzewski
Abstract.
This study aimed to determine the efficacy of low molecular weight heparin (LMWH) and lidocaine combined with LMWH for improving the course of acute pancreatitis (AP). A total of 30 rats were divided into three groups: the NaCl group, which received an intraarterial infusion of 0.9% sodium chloride; the Heparin group, which received a subcutaneous injection of LMWH; and the Lidocaine–Heparin group, which received an intraarterial infusion of 1% lidocaine, with subsequent subcutaneous injection of LMWH. AP was triggered using 80 μg/kg body weight of cerulein. Serum amylase and lipase levels were evaluated before induction of AP (measurement 0 – M0), after triggering AP (measurement 1 – M1), 1 h (measurement 2 – M2), 3 h (measurement 3 – M3), and 5 h (measurement 4 – M4) after treatment. After euthanasia, pancreatic tissues were collected for pathological analysis. No intergroup differences in serum amylase and lipase levels were observed between the NaCl and Heparin groups in all post-treatment evaluation points (M2, M3, and M4). Conversely, the Lidocaine–Heparin group showed significantly lower amylase values than the NaCl and Heparin groups in all post-treatment evaluation points. Furthermore, the Lidocaine–Heparin group showed significantly lower lipase values compared with the NaCl group in the first post-treatment evaluation point (M2), as well as compared with the Heparin group in the first (M2) and second (M3) post-treatment evaluation points. No significant intergroup differences were observed in pathological pancreatic tissue evaluation. Subcutaneous injection of LMWH did not impact the natural course of AP. However, the addition of intraarterially administered 1% lidocaine solution significantly reduced the severity of AP.
Keywords: Experimental • Microcirculation • continuous regional arterial infusion • Vasodilation • Anticoagulation
Semaphorin 5A as a Novel Diagnostic Biomarker of Juvenile Idiopathic Arthritis
Justyna Roszkiewicz · Krystyna Wyka · Elżbieta Smolewska
Abstract.
Although juvenile idiopathic arthritis (JIA) is the most common form of autoimmune-driven arthritis in pediatric population, due to the lack of universal diagnostic markers and heterogeneity of the disease, many patients face a significant delay when establishing the diagnosis. As the hallmark of JIA is the inflammation of synovial membrane with pathological angiogenesis, this study aimed to compare the concentrations of semaphorin 5A (SEMA5A), protein involved in both angiogenesis and immune response, between new-onset JIA and healthy controls. Thirty-five JIA patients and 35 sex-and-age matched healthy controls were enrolled in this study. Serum concentrations of SEMA5A and vascular endothelial growth factor A (VEGF-A) were established using the enzyme-linked immunosorbent assay (ELISA) method. The serum concentration of SEMA5A was elevated in JIA patients in comparison to healthy controls with the median value of 2.04 (interquartile range [IQR]: 12.41) in JIA patients and 1.34 (IQR: 1.79) in healthy controls (p = 0.002). The difference was particularly prominent in oligoarticular
JIA, where median SEMA5A concentration equaled 1.76 ng/mL (IQR: 1.56) in comparison to 0.33 ng/mL (IQR: 1.34) in sex-and-age-matched healthy controls (p = 0.001). SEMA5A concentration correlated strongly with VEGF-A concentration (r = 0.807, p < 0.001) and differed significantly in subgroups of different synovial membrane power-Doppler ultrasound (PDUS) inflammatory activities (p = 0.018). Although our findings need to be repeated on larger groups of JIA patients, SEMA5A is a promising marker of JIA, linked to pathological angiogenesis of synovial membrane in this entity. Moreover, it may be useful as a target of innovative treatment strategies in the nearest future.
Keywords: Juvenile idiopathic arthritis • Oligoarticular juvenile idiopathic arthritis • Angiogenesis • Semaphorins • Semaphorin 5A • SEMA5A
TAGLN2 Exacerbates Acute Pancreatitis-Induced Liver Injury by Increasing Hepatocyte Pyroptosis via Kupffer Cells-Mediated Inflammatory Response
Huigeng Zhao · Yalan Luo · Xi Chen · Ju Wu · Zizhao Zhu · Hailong Chen
Abstract.
Pyroptosis, a programmed form of inflammatory cell death, has been demonstrated to participate in both Acute pancreatitis (AP) and its complication liver injury. Transgelin-2 (TAGLN2), an actin-binding protein involved in inflammatory response, has been reported to be highly expressed in AP. However, the role of TAGLN2 in AP-induced liver injury remains unclear. Mice were treated with cerulein to construct the AP model in vivo, while Kupffer cells were stimulated with lipopolysaccharide (LPS) to mimic in vitro model. A series of in vitro and in vivo experiments were performed to investigate the role and mechanism of TAGLN2 in AP-induced liver injury. Cerulein administration induced pathological injury of the pancreatic and liver tissues, along with elevated levels of amylase, lipase, alanine aminotransferase (ALT), and aspartate transaminase (AST). TAGLN2 was significantly elevated at both the transcriptional and translational levels in the hepatocytes and Kupffer cells of AP mice. Knockout of TAGLN2 alleviated liver injury by reducing inflammatory cytokine levels, pyroptosis-related protein expression, and liver
dysfunction markers. The relative levels of inflammatory factors, the expressions of pyroptosis-related proteins, and the pyroptosis rate were increased in LPSinduced Kupffer cells in an in vitro model, whereas TAGLN2 knockdown reversed these changes. Mechanistically, TAGLN2 promoted activation of the ANXA2/ NF-κB axis in Kupffer cells, contributing to the inflammatory response. TAGLN2 exacerbates AP-induced liver injury by enhancing hepatocyte pyroptosis through Kupffer cell-mediated inflammatory activation of the ANXA2/ NF-κB axis. Targeting TAGLN2 may offer a potential therapeutic strategy for mitigating liver injury in AP.
Keywords: Liver injury • Acute pancreatitis • TAGLN2 • Pyroptosis • Kupffer cells • Inflammatory response
Impact of Intraoperative Cell Salvage on Circulating Tumor Cells and Cellular Activity in Patients with Hepatocellular Carcinoma Undergoing Curative Resection: A Prospective Comparative Study
Ke Yue, Lai-wei You, Bao-ji Hu, Yong Cheng, Jinhuo Wang, Hao Li, Jianrong Guo
Abstract.
This study investigates the effects of intraoperative cell salvage (IOCS) on cell survival rates, apoptosis levels, and circulating tumor cell (CTC) counts in patients with hepatocellular carcinoma (HCC) undergoing curative resection. A combination of immunofluorescence, Western blot, flow cytometry, and qRT-PCR was employed to assess the impact of IOCS on cellular activity and CTC dynamics in these patients. No significant differences were found in demographic characteristics, including gender, age, body mass index (BMI), Child-Pugh classification, and liver function markers (alanine aminotransferase [ALT] and aspartate aminotransferase [AST]) between the experimental and control groups. Preoperatively, both groups exhibited low cell survival rates without statistical differences (P > 0.05), and cell survival remained similarly low during surgery. However, 6 h post-surgery, the experimental group showed a significant increase in cell survival rates compared with the control group (P < 0.05), suggesting that IOCS enhances postoperative cell viability. Apoptosis levels were similarly high in both groups before and during surgery (P > 0.05), but notably, 6 h post-operation, apoptosis levels in the experimental group were significantly reduced (P < 0.05), indicating effective prevention of cell death. Although preoperative CTC counts were low and increased during surgery, no significant differences were observed between groups during surgery. However, 6 h post-surgery, the experimental group exhibited a marked decrease in CTC counts (P < 0.05), indicating a reduction in tumor cell dissemination. Although these findings suggest that IOCS improves cell survival and reduces apoptosis and CTC counts, there is a potential concern regarding the possibility of IOCS preferentially preserving viable cells with metastatic potential. The long-term impact of this intervention on tumor recurrence or metastasis requires further investigation. In conclusion, IOCS appears to offer short-term benefits in enhancing postoperative cell survival and reducing CTC dissemination in patients with HCC; however, the potential risk of promoting metastatic cell viability warrants additional study before broader clinical application.
Keywords: Intraoperative cell salvage • Circulating tumor cell • Hepatocellular carcinoma • Cellular activity
High Expression of CIP2A Can Promote the Proliferation, Migration, and Epithelial-Mesenchymal Transition of Diffuse Large B-Cell Lymphoma Cells
Caifang Zhao · Xiang Weng · Wei He · Yanming Lei
Abstract.
Diffuse large B-cell lymphoma (DLBC) is one of the usual forms found in indolent or invasive non-Hodgkin’s lymphoma. Cancerous inhibitor of protein phosphatase 2A (CIP2A) has been revealed to be dysregulated in multiple cancers and is closely associated with tumor growth. However, the regulatory influences of CIP2A in DLBC progression remain unclear. The protein expressions were determined through western blot. Cell survival was assessed through the CCK-8 assay. Cell proliferation was examined through colony formation assay. The cell migration and invasion were inspected through transwell assay. First, it was discovered that CIP2A exhibited higher expression in DLBC. Additionally, inhibition of CIP2A restrained cell growth and metastasis in DLBC. Next, it was discovered that E-cadherin protein expression was ascended as well as N-cadherin and α-SMA protein expressions were descended after CIP2A knockdown, indicating that CIP2A suppression can retard the epithelial-mesenchymal transition (EMT) progress in DLBC. Finally, it was demonstrated that suppression of CIP2A retarded the Wnt/β-catenin pathway. It was manifested that high expression of CIP2A can aggrandize cell proliferation, migration, and EMT process in DLBC, and triggered the Wnt/β-catenin pathway. This finding implied that CIP2A may serve as a hopeful target for treating DLBC.
Keywords: CIP2A • EMT process • Wnt/β-catenin pathway • Diffuse large B-cell lymphoma
Autoimmune Diseases and Pregnancy – Is There a Greater Risk of Giving Birth to a Sick Child?
Magdalena Pszczołowska · Kamil Walczak · Weronika Kołodziejczyk · Magdalena Mroziak · Gracjan Kozłowski · Jerzy Leszek
Abstract.
This study aims to describe the impact of some of the most common autoimmune diseases, such as multiple sclerosis (MS) and neuromyelitis optica spectrum disorders (NMOSD), myasthenia gravis (MG), chronic inflammatory bowel disease (IBD), type 1 diabetes (T1D), autoimmune thyroid disease, rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), psoriasis, vasculitis, and antiphospholipid syndrome (APS), on pregnancy in women. This review investigates the risk to the offspring of the women; that is, whether the mother’s disease may affect fertility or disturb fetal development.
Keywords: Autoimmune disease • Pregnancy • Pregnant woman • Multiple sclerosis • IBD • TD1
The Role of Glomerular and Serum Expression of Lymphocyte Activating Factors BAFF and APRIL in Patient with Membranous and IgA Nephropathies
Barbara Moszczuk · Krzysztof Mucha · Róża Kucharczyk · Radosław Zagożdżon
Abstract.
Increased activity of B lymphocytes underpins many autoimmune conditions. A key component of the humoral immune response involves the A PRoliferation-Inducing Ligand (APRIL) and B-cell-activating factor (BAFF) system. These proteins are responsible for the activation, maturation, and survival of B lymphocytes, playing a pivotal role in autoimmunity. Therefore, targeting the BAFF/APRIL system proves promising for the treatment of various autoimmune diseases. Meticulous research into pathomechanisms of lupus nephritis (LN) has enabled the introduction of biological treatments targeting the BAFF-mediated pathway, significantly improving prognosis. In certain types of glomerulonephritis (GN), increased levels of the BAFF/APRIL system might be associated with higher proteinuria, elevated serum creatinine, but also with specific histopathological features. This indicates that biological therapies currently available could be repurposed for conditions where increased activation of B lymphocytes plays a critical role in the disease’s pathophysiology. Understanding the mechanisms underlying autoimmune diseases will facilitate the adaptation of novel drugs for orphan diseases. That is why the use of chimeric antigen receptor T (CAR-T) cells as agents against B-cells receptor (BCR), represents a highly targeted and potentially optimal treatment approach. This study summarizes current knowledge about the role of the BAFF/APRIL system in lymphocyte activation mechanisms, particularly in GN. It also discusses existing biological treatments and explores future directions for drug development based on the CAR-T cell technology.
Keywords: APRIL • autoimmunity • BAFF • glomerulonephritis • IgA nephropathy • membranous nephropathy
RFWD3 Knockdown Inhibits Cancer Cell Proliferation, Migration, and Invasion while Promoting Apoptosis of Non-Small Cell Lung
Cancer Cells
Ting Ma · Baolan Wang
Abstract.
RING finger and WD repeat domain 3 (RFWD3) expression is elevated in various tumor types, but its precise role in non-small cell lung cancer (NSCLC) remains unclear. This study aims to investigate the biological function of RFWD3 in NSCLC and the associated molecular pathways. RFWD3 expression was knocked down in NSCLC cells through transfection. Cell apoptosis was analyzed using flow cytometry, cell viability was assessed using the cell counting kit-8 (CCK-8), and the migration and invasion of NSCLC cells were evaluated using Transwell chamber assays. Additionally, the expression of BAX, BCL-2, cleaved-caspase-3, and key signaling molecules involved in the ERK/p38 pathway was determined using Western blotting. The expression of RFWD3 was found to be elevated in NSCLC cells compared with normal lung epithelial BEAS-2B cells. Its knockdown led to reduced cell proliferation, migration, and invasion and increased apoptosis rate, partially through the inhibition of the ERK/p38 signaling pathway. Knockdown of the RFWD3 gene inhibited NSCLC cell proliferation, migration, and invasion while also inducing apoptosis. These effects are partially attributed to the blockade of the ERK/p38 signaling pathway.
Keywords: non-small cell lung cancer • RFWD3 • apoptosis • migration and invasion • ERK/p38
IL-17B Inhibits Hepatocellular Carcinoma Cell Proliferation
Joanna Pastwińska · Iwona Karwaciak · Kaja Karaś · Daria Grabarczyk · Anna Sałkowska · Marcin Ratajewski
Abstract.
IL-17RB is a cytokine receptor that binds interleukin (IL)-17B and IL-17E. While analyzing IL-17RB expression in various cancer cell lines, we found that this receptor is highly expressed at both the mRNA and protein levels in hepatocellular carcinoma (HCC) cells. This finding prompted us to investigate the effects of its ligand, IL-17B, on the proliferation of these cells. Our results demonstrate that IL-17B inhibits the proliferation of HCC cells through an AKT-dependent, but NF-κB-independent, mechanism. Additionally, IL-17B affected colony formation in these cells. Interestingly, these effects were not observed in melanoma cells, which express low levels of IL-17RB. Our results may represent a promising new approach for treating HCC – a disease for which immunological therapies have recently gained significant attention – especially considering that HCC patients experience progressive liver dysfunction and that the IL-17B–IL-17RB signaling pathway plays a role in liver regeneration.
Keywords: IL-17B • IL-17RB • Hepatocellular carcinoma • Proliferation
Knockdown of MBD2 Attenuates LPS-Stimulated Inflammation and Apoptosis in WI-38 Cells Through the STAT-3 Pathway
Yao Chen · Liqun Lu
Abstract.
This study aims to investigate the role of MBD protein 2 (MBD2) in the pneumonia cell model of lipopolysaccharide (LPS)-stimulated WI-38 cells and to uncover the mechanism. LPS-stimulated WI-38 cells were constructed as an in vitro pneumonia model. Quantitative polymerase chain reaction (qPCR) and immunoblot assays showed MBD2 expression in WI-38 cells. Cell counting kit-8 (CCK-8) assays showed the growth of WI-38 cells. Flow cytometry assays showed the apoptosis of WI-38 cells after LPS treatment and siRNA transfection. Enzyme-linked immunosorbent assay (ELISA) and qPCR assays showed the effects on inflammation, and immunoblot assays further confirm the mechanism. MBD2 was highly expressed in LPS-stimulated WI-38 cells. Knockdown of MBD2 alleviates production of cellular inflammatory cytokines in LPS-stimulated WI-38 cells. Further, knockdown of MBD2 alleviates apoptosis in LPS-stimulated WI-38 cells. Mechanically, the knockdown of MBD2 regulates the signal transducer and activator of transcription (STAT)-3 pathway in LPS-stimulated WI-38 cells. Knockdown of MBD2 attenuates LPS-stimulated inflammation and apoptosis in WI-38 cells through the STAT-3 pathway. Therefore, MBD2 could serve as a promising target of pediatric pneumonia.
Keywords: Pneumonia • MBD protein 2 • Inflammatory cytokines • Apoptosis • STAT-3
Overexpression of Annexin A1 Inhibits Pyroptosis and Improves Dry Eye Signs by Regulating the TRIM72/Nrf2/HO-1 Signaling Pathway
Li Zhang · Peng Chen · Pengfei Han · Huizhe Fu · Bin Sun
Abstract.
This study aimed to investigate the therapeutic potential of annexin A1 (ANXA1) overexpression in improving the signs of dry eye disease (DED) and to elucidate the underlying molecular mechanism. A murine model of DED was established by topical application of 0.2% benzalkonium chloride (BAC), and human corneal epithelial (HCE-T) cells were exposed to 0.0005% BAC for in vitro experiments. ANXA1 was overexpressed using adenoviral vectors, and the effects on tear production, pyroptosis, and activation of the tripartite motif-containing protein 72 (TRIM72)/nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway were evaluated using Western blotting, Schirmer test, Terminal deoxynucleotidyl Transferase dUTP Nick End Labeling (TUNEL) assays, and fluorescent probe analyses. To further examine the role of TRIM72, its expression was silenced with specific small interfering RNA (siRNA), and the consequent impact on ANXA1-mediated therapeutic effects was assessed. ANXA1 expression was significantly reduced in both in vivo and in vitro DED models. Restoration of ANXA1 through overexpression significantly improved tear secretion and suppressed pyroptosis in the murine model. Similarly, in HCE-T cells, ANXA1 overexpression not only enhanced cellular proliferation but also significantly inhibited pyroptosis. Mechanistic investigations demonstrated that ANXA1 overexpression activated the TRIM72/Nrf2/HO-1 signaling pathway by increasing TRIM72, Nrf2, and HO-1 expression. Notably, silencing TRIM72 abolished the therapeutic effects of ANXA1 overexpression, thereby confirming that activation of this pathway is essential for mediating the protective effects of ANXA1 against DED. Overexpression of ANXA1 inhibits pyroptosis and improves dry eye signs by regulating the TRIM72/Nrf2/HO-1 axis.
Keywords: Dry eye disease • ANXA1 • Pyroptosis • TRIM72 • Nrf2
This retraction concerns the following article:
DOI: 10.1007/s00005-006-0019-x
„Modulation of apoptosis signaling for cancer therapy”, Arch Immunol Ther Exp 54, 173-175 (2006) by Simone Fulda, Klaus-Michael Debatin.
A significant overlap (78%) occurs between the mentioned above article and the article entitled „Apoptosis in drug response”, Current Pharmacogenomics 1,
9-16 (2003), by the same authors, who did not respond to our request for comments. The article is therefore retracted.
From Fleas to T Lymphocytes and Beyond: How did I Become an Immunologist, and What Has Resulted from It?
Paweł Kisielow
Abstract.
A narrative from a co-author of studies explaining the cellular mechanisms of self/nonself discrimination by the immune system, set against the backdrop of significant advancements in immunology during the last quarter of the 20th century. This review focuses on demonstrating the functional diversity of T cells by identifying their two major subsets: regulatory CD4 and cytotoxic CD8. It also examines the first T-cell receptor (TCR) transgenic mice, providing definitive evidence for both positive and negative selection of immature thymocytes as mechanisms for generating a self-tolerant and MHC-restricted TCR repertoire. The origins of these experiments have been discussed.
Keywords: Self/nonself discrimination • Central tolerance • Positive selection • MHC-restriction
Significance of the LL-37 Peptide Delivered from Human Cathelicidin in the Pathogenesis, Treatment, and Diagnosis of Sepsis
Angelika Mańkowska, · Paulina Paprocka, · Grzegorz Król, · Agata Lesiak, · Jakub Spałek, · Ewelina Piktel · Sławomir Okła · Piotr Bijak · Wiesława Niklińska · Bonita Durnaś · Robert Bucki
Abstract.
Antimicrobial peptides, which function as the first line of host immune defense, have recently been identified as important immunomodulators of inflammation, and are involved as regulatory molecules in infections, including sepsis. Treatment of sepsis is very complex and remains largely challenging and sometimes ineffective. This creates a need to develop new therapeutic strategies focusing not only on the elimination of sepsis-causing microorganisms, which can be achieved with antibiotics, but also on the control of the immune system and its overactive response resulting in increased vascular endothelial permeability. One approach to develop new treatments for patients with sepsis is to better understand the pleiotropic function of the human LL-37 peptide that originates from the human cathelicidin antibacterial protein (h-CAP18). An increasing number of studies indicate high dynamics of changes in LL-37 concentration in the blood during sepsis. Additionally, in animal models, administration of exogenous LL-37 peptide to mice with experimentally induced sepsis increases their survival. It can therefore be assumed that knowledge of the molecular mechanism of cathelicidin LL-37 action, as well as the synthesis of its stable analogs, will result in progress in the diagnosis and therapy of sepsis.
Keywords: Sepsis • Cathelicidins • LL-37 peptide • Bacteria