Biomedicines

23 pages, 599 KiB

Open AccessReview

Sex and Gender Influences on Cancer Immunotherapy Response

by Azzurra Irelli, Maria Maddalena Sirufo, Carlo D’Ugo, Lia Ginaldi and Massimo De Martinis

Biomedicines 2020, 8(7), 232; https://doi.org/10.3390/biomedicines8070232 - 21 Jul 2020

Cited by 73 | Viewed by 6461

The global burden of cancer is growing and a wide disparity in the incidence, malignancy and mortality of different types of cancer between each sex has been demonstrated. The sex specificity of cancer appears to be a relevant issue in the management of [...] Read more.

The global burden of cancer is growing and a wide disparity in the incidence, malignancy and mortality of different types of cancer between each sex has been demonstrated. The sex specificity of cancer appears to be a relevant issue in the management of the disease, and studies investigating the role of sex and gender are becoming extremely urgent. Sex hormones are presumably the leading actors of sex differences in cancer, especially estrogens. They modulate gene expression, alter molecules and generate disparities in effectiveness and side effects of anticancer therapies. Recently immunotherapy aims to improve anticancer treatment strategies reducing off-target effects of chemotherapy and direct cancer cells killing. It is recognized as a fruitful strategy to treat and possible to cure cancer. Immunotherapeutic agents are used to activate or boost the activation of the immune system to fight cancer cells through physiological mechanisms often evaded in the offensive march of the disease. These therapeutic strategies have allowed new successes, but also have serious adverse effects including non-specific inflammation and autoimmunity. Sex and gender issues are of primary importance in this field, due to their recognized role in inflammation, immunity and cancer, and the clarification and understanding of these aspects is a necessary step to increase the responses and to diminish the adverse effects of immunotherapy. This review describes the available knowledge on the role of sex and gender in cancer immunotherapy, and will offer insights to stimulate the attention and practice of clinicians and researchers in a gender perspective of new cancer treatment strategies. Full article

(This article belongs to the Section Immunology and Immunotherapy)

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16 pages, 1183 KiB

Open AccessReview

Asthma and Obesity in Children

by Francesco Sansone, Marina Attanasi, Sabrina Di Pillo and Francesco Chiarelli

Biomedicines 2020, 8(7), 231; https://doi.org/10.3390/biomedicines8070231 - 21 Jul 2020

Cited by 30 | Viewed by 7286

Abstract

Asthma and obesity are two major chronic diseases in children and adolescents. Recent scientific evidence points out a causative role of obesity in asthma predisposition. However, studies assessing the real impact of excessive weight gain on lung function in children have shown heterogeneous [...] Read more.

Asthma and obesity are two major chronic diseases in children and adolescents. Recent scientific evidence points out a causative role of obesity in asthma predisposition. However, studies assessing the real impact of excessive weight gain on lung function in children have shown heterogeneous results. In this review, the pathological mechanisms linking obesity and development of asthma in children are summarized and factors influencing this relationship are evaluated. Common disease modifying factors including age, sex, ethnicity, development of atopic conditions, and metabolic alterations significantly affect the onset and phenotypic characteristics of asthma. Given this, the impact of these several factors on the obesity–asthma link were considered, and from revision of the literature we suggest the possibility to define three main clinical subtypes on the basis of epidemiological data and physiological–molecular pathways: obese-asthmatic and atopy, obese-asthmatic and insulin-resistance, and obese-asthmatic and dyslipidemia. The hypothesis of the different clinical subtypes characterizing a unique phenotype might have an important impact for both future clinical management and research priorities. This might imply the necessity to study the obese asthmatic child with a “multidisciplinary approach”, evaluating the endocrinological and pneumological aspects simultaneously. This different approach might also make it possible to intervene earlier in a specific manner, possibly with a personalized and tailored treatment. Surely this hypothesis needs longitudinal and well-conducted future studies to be validated. Full article

(This article belongs to the Section Molecular and Translational Medicine)

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18 pages, 1651 KiB

Open AccessReview

Antiplatelet Therapy for Acute Respiratory Distress Syndrome

by Chuan-Mu Chen, Hsiao-Ching Lu, Yu-Tang Tung and Wei Chen

Biomedicines 2020, 8(7), 230; https://doi.org/10.3390/biomedicines8070230 - 21 Jul 2020

Cited by 21 | Viewed by 13864

Abstract

Acute respiratory distress syndrome (ARDS) is a common and devastating syndrome that contributes to serious morbidities and mortality in critically ill patients. No known pharmacologic therapy is beneficial in the treatment of ARDS, and the only effective management is through a protective lung [...] Read more.

Acute respiratory distress syndrome (ARDS) is a common and devastating syndrome that contributes to serious morbidities and mortality in critically ill patients. No known pharmacologic therapy is beneficial in the treatment of ARDS, and the only effective management is through a protective lung strategy. Platelets play a crucial role in the pathogenesis of ARDS, and antiplatelet therapy may be a potential medication for ARDS. In this review, we introduce the overall pathogenesis of ARDS, and then focus on platelet-related mechanisms underlying the development of ARDS, including platelet adhesion to the injured vessel wall, platelet-leukocyte-endothelium interactions, platelet-related lipid mediators, and neutrophil extracellular traps. We further summarize antiplatelet therapy, including aspirin, glycoprotein IIb/IIIa receptor antagonists, and P2Y12 inhibitors for ARDS in experimental and clinical studies and a meta-analysis. Novel aspirin-derived agents, aspirin-triggered lipoxin, and aspirin-triggered resolvin D1 are also described here. In this narrative review, we summarize the current knowledge of the role of platelets in the pathogenesis of ARDS, and the potential benefits of antiplatelet therapy for the prevention and treatment of ARDS. Full article

(This article belongs to the Special Issue Oxidative Stress and Inflammation: From Mechanisms to Therapeutic Approaches)

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16 pages, 590 KiB

Open AccessReview

Depression and Cognitive Impairment—Extrahepatic Manifestations of NAFLD and NASH

by Martina Colognesi, Daniela Gabbia and Sara De Martin

Biomedicines 2020, 8(7), 229; https://doi.org/10.3390/biomedicines8070229 - 21 Jul 2020

Cited by 67 | Viewed by 7682

Abstract

Non-alcoholic fatty liver disease (NAFLD) and its complication non-alcoholic steatohepatitis (NASH) are important causes of liver disease worldwide. Recently, a significant association between these hepatic diseases and different central nervous system (CNS) disorders has been observed in an increasing number of patients. NAFLD-related [...] Read more.

Non-alcoholic fatty liver disease (NAFLD) and its complication non-alcoholic steatohepatitis (NASH) are important causes of liver disease worldwide. Recently, a significant association between these hepatic diseases and different central nervous system (CNS) disorders has been observed in an increasing number of patients. NAFLD-related CNS dysfunctions include cognitive impairment, hippocampal-dependent memory impairment, and mood imbalances (in particular, depression and anxiety). This review aims at summarizing the main correlations observed between NAFLD development and these CNS dysfunctions, focusing on the studies investigating the mechanism(s) involved in this association. Growing evidences point at cerebrovascular alteration, neuroinflammation, and brain insulin resistance as NAFLD/NASH-related CNS manifestations. Since the pharmacological options available for the management of these conditions are still limited, further studies are needed to unravel the mechanism(s) of NAFLD/NASH and their central manifestations and identify effective pharmacological targets. Full article

(This article belongs to the Special Issue NASH and Systemic Complications: From Basic to Clinical Research)

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19 pages, 3618 KiB

Open AccessReview

Novel MRI Techniques Identifying Vascular Leak and Paravascular Flow Reduction in Early Alzheimer Disease

by Charles R Joseph

Biomedicines 2020, 8(7), 228; https://doi.org/10.3390/biomedicines8070228 - 20 Jul 2020

Cited by 15 | Viewed by 4283

Abstract

With beta amyloid and tau antibody treatment trial failures, avenues directed to other facets of the disease pathophysiology are being explored to treat in the preclinical or early clinical state. Clear evidence of blood–brain barrier (BBB) breakdown occurring early in the AD process [...] Read more.

With beta amyloid and tau antibody treatment trial failures, avenues directed to other facets of the disease pathophysiology are being explored to treat in the preclinical or early clinical state. Clear evidence of blood–brain barrier (BBB) breakdown occurring early in the AD process has recently been established. Likewise, the glymphatic system regulating water and solute inflow and outflow in parallel with the vascular system is affected causing delayed clearance of fluid waste. Its dysfunction as a component of AD along with BBB leak are reasonable candidates to explore for future treatments. Ideally, human medication trials require a minimally invasive method of quantifying both improvements in BBB integrity and glymphatic fluid clearance correlated with clinical outcomes. We will review the known physiology and anatomy of the BBB system, and its relationship to the glymphatic system and the microglial surveillance system. Dysfunction of this tripart system occurring in preclinical Alzheimer disease (AD) will be reviewed along with existing MRI tools for identifying altered flow dynamics useful for monitoring improved functionality with future treatments. High-resolution dynamic contrast enhanced MRI imaging demonstrating BBB leak and the recently reported non-invasive 3D PASL MRI pilot study demonstrating significant delay in glymphatic clearance in AD subjects appear to be the best candidates. Full article

(This article belongs to the Special Issue Alzheimer Disease: Controversies in Basic Science Research, Different Theories, and Reasons for Failed Trials)

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24 pages, 2153 KiB

Open AccessEditor’s ChoiceReview

Sphingosine-1-Phosphate: Its Pharmacological Regulation and the Treatment of Multiple Sclerosis: A Review Article

by Stanley Cohan, Elisabeth Lucassen, Kyle Smoot, Justine Brink and Chiayi Chen

Biomedicines 2020, 8(7), 227; https://doi.org/10.3390/biomedicines8070227 - 18 Jul 2020

Cited by 41 | Viewed by 6162

Abstract

Sphingosine-1-phosphate (S1P), via its G-protein-coupled receptors, is a signaling molecule with important regulatory properties on numerous, widely varied cell types. Five S1P receptors (S1PR1-5) have been identified, each with effects determined by their unique G-protein-driven downstream pathways. The discovery that lymphocyte egress from [...] Read more.

Sphingosine-1-phosphate (S1P), via its G-protein-coupled receptors, is a signaling molecule with important regulatory properties on numerous, widely varied cell types. Five S1P receptors (S1PR1-5) have been identified, each with effects determined by their unique G-protein-driven downstream pathways. The discovery that lymphocyte egress from peripheral lymphoid organs is promoted by S1P via S1PR-1 stimulation led to the development of pharmacological agents which are S1PR antagonists. These agents promote lymphocyte sequestration and reduce lymphocyte-driven inflammatory damage of the central nervous system (CNS) in animal models, encouraging their examination of efficacy in the treatment of multiple sclerosis (MS). Preclinical research has also demonstrated direct protective effects of S1PR antagonists within the CNS, by modulation of S1PRs, particularly S1PR-1 and S1PR-5, and possibly S1PR-2, independent of effects upon lymphocytes. Three of these agents, fingolimod, siponimod and ozanimod have been approved, and ponesimod has been submitted for regulatory approval. In patients with MS, these agents reduce relapse risk, sustained disability progression, magnetic resonance imaging markers of disease activity, and whole brain and/or cortical and deep gray matter atrophy. Future opportunities in the development of more selective and intracellular S1PR-driven downstream pathway modulators may expand the breadth of agents to treat MS. Full article

(This article belongs to the Special Issue Multiple Sclerosis: Diagnosis and Treatment II)

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19 pages, 3221 KiB

Open AccessArticle

Increased IL-2 and Reduced TGF-β Upon T-Cell Stimulation are Associated with GM-CSF Upregulation in Multiple Immune Cell Types in Multiple Sclerosis

by Jehan Aram, Nanci Frakich, Elena Morandi, Mohammed Alrouji, Amal Samaraweera, David Onion, Ian Spendlove, Sergio L. Colombo, Radu Tanasescu, Bruno Gran and Cris S. Constantinescu

Biomedicines 2020, 8(7), 226; https://doi.org/10.3390/biomedicines8070226 - 18 Jul 2020

Cited by 7 | Viewed by 4848

Abstract

Granulocyte macrophage colony stimulating factor (GM-CSF) is a pro-inflammatory cytokine produced by immune cells. Recent evidence suggests that GM-CSF plays an important role in multiple sclerosis (MS) pathogenesis. We investigated the expression and regulation of GM-CSF in different immune cells in MS. We [...] Read more.

Granulocyte macrophage colony stimulating factor (GM-CSF) is a pro-inflammatory cytokine produced by immune cells. Recent evidence suggests that GM-CSF plays an important role in multiple sclerosis (MS) pathogenesis. We investigated the expression and regulation of GM-CSF in different immune cells in MS. We also investigated the differentiation and frequency of GM-CSF-producing Th cells that do not co-express interferon (IFN)-γ or interleukin-17 (IL-17) (Th-GM cells) in MS. We found a significant increase in the percentage of GM-CSF-expressing Th cells, Th1 cells, Th-GM cells, cytotoxic T (Tc) cells, monocytes, natural killer (NK) cells, and B cells in PBMC from MS patients stimulated with T cell stimuli. Stimulated PBMC culture supernatants from MS patients contained significantly higher levels of IL-2, IL-12, IL-1β, and GM-CSF and significantly lower levels of transforming growth factor (TGF-)β. Blocking IL-2 reduced the frequency of Th-GM cells in PBMC from MS patients. The frequency of Th-GM cells differentiated in vitro from naïve CD4⁺ T cells was significantly higher in MS patients and was further increased in MS with IL-2 stimulation. These findings suggest that all main immune cell subsets produce more GM-CSF in MS after in vitro stimulation, which is associated with defective TGF-β and increased IL-2 and IL-12 production. Th-GM cells are increased in MS. GM-CSF may be a potential therapeutic target in MS. Full article

(This article belongs to the Special Issue Immune Mediated Inflammatory Diseases: From an Update on Molecular and Cellular Mechanisms to Biomarkers for Personalized Medicine)

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12 pages, 1447 KiB

Open AccessArticle

Pharmacological Effects of a Novel Bradykinin-Related Peptide (RR-18) from the Skin Secretion of the Hejiang Frog (Ordorrana hejiangensis) on Smooth Muscle

by Xiaowei Zhou, Jie Xu, Ruimin Zhong, Chengbang Ma, Mei Zhou, Zhijian Cao, Xinping Xi, Chris Shaw, Tianbao Chen, Lei Wang and Hang Fai Kwok

Biomedicines 2020, 8(7), 225; https://doi.org/10.3390/biomedicines8070225 - 17 Jul 2020

Cited by 3 | Viewed by 2918

Abstract

Bradykinin (BK) and bradykinin-related peptides (BRPs), which were identified from a diversity of amphibian skin secretions, exerted contractile and relaxing effects on non-vascular and vascular smooth muscle, respectively. Here, we report a novel bradykinin-related peptide with a molecular mass of 1890.2 Da, RVAGPDKPARISGLSPLR, [...] Read more.

Bradykinin (BK) and bradykinin-related peptides (BRPs), which were identified from a diversity of amphibian skin secretions, exerted contractile and relaxing effects on non-vascular and vascular smooth muscle, respectively. Here, we report a novel bradykinin-related peptide with a molecular mass of 1890.2 Da, RVAGPDKPARISGLSPLR, which was isolated and identified from Ordorrana hejiangensis skin secretions, followed by a C-terminal extension sequence VAPQIV. The biosynthetic precursor-encoding cDNA was cloned by the “shotgun” cloning method, and the novel RR-18 was identified and structurally confirmed by high-performance liquid chromatography (HPLC) and tandem mass spectrometry (MS/MS). Subsequently, the myotropic activity of the synthetic replicate of RR-18 was investigated on the rat bladder, uterus, tail artery and ileum smooth muscle. The peptide was named RR-18 in accordance (R = N-terminal arginine, R = C-terminal arginine, 18 = number of residues). In this study, the synthetic replicates of RR-18 showed no agonist/antagonism of BK-induced rat bladder and uterus smooth muscle contraction. However, it displayed an antagonism of bradykinin-induced rat ileum contraction and arterial smooth muscle relaxation. The EC₅₀ values of BK for ileum and artery, were 214.7 nM and 18.3 nM, respectively. When the tissue was pretreated with the novel peptide, RR-18, at the maximally effective concentration of bradykinin (1 × 10⁻⁶ M), bradykinin-induced contraction of the ileum and relaxation of the arterial smooth muscle was reduced by 50–60% and 30–40%, respectively. In conclusion, RR-18 represents novel bradykinin antagonising peptide from amphibian skin secretions. It may provide new insight into possible treatment options for chronic pain and chronic inflammation. Full article

(This article belongs to the Special Issue Animal Venoms–Curse or Cure?)

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20 pages, 5599 KiB

Open AccessReview

HDX-MS: An Analytical Tool to Capture Protein Motion in Action

by Dominic Narang, Cristina Lento and Derek J. Wilson

Biomedicines 2020, 8(7), 224; https://doi.org/10.3390/biomedicines8070224 - 17 Jul 2020

Cited by 38 | Viewed by 9874

Abstract

Virtually all protein functions in the cell, including pathogenic processes, require coordinated motion of atoms or domains, i.e., conformational dynamics. Understanding protein dynamics is therefore critical both for drug development and to learn about the underlying molecular causes of many diseases. Hydrogen–Deuterium Exchange [...] Read more.

Virtually all protein functions in the cell, including pathogenic processes, require coordinated motion of atoms or domains, i.e., conformational dynamics. Understanding protein dynamics is therefore critical both for drug development and to learn about the underlying molecular causes of many diseases. Hydrogen–Deuterium Exchange Mass Spectrometry (HDX-MS) provides valuable information about protein dynamics, which is highly complementary to the static picture provided by conventional high-resolution structural tools (i.e., X-ray crystallography and structural NMR). The amount of protein required to carry out HDX-MS experiments is a fraction of the amount required by alternative biophysical techniques, which are also usually lower resolution. Use of HDX-MS is growing quickly both in industry and academia, and it has been successfully used in numerous drug and vaccine development efforts, with important roles in understanding allosteric effects and mapping binding sites. Full article

(This article belongs to the Special Issue Protein Structure, Function and Dynamics in Diseases and Therapeutics)

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12 pages, 877 KiB

Open AccessReview

Key Milestones Contributing to the Understanding of the Mechanisms Underlying Fibromyalgia

by Geoffrey Littlejohn and Emma Guymer

Biomedicines 2020, 8(7), 223; https://doi.org/10.3390/biomedicines8070223 - 17 Jul 2020

Cited by 20 | Viewed by 7679

Abstract

The promulgation of the American College of Rheumatology (ACR) 1990 criteria for fibromyalgia (FM) classification has significantly contributed to an era of increased research into mechanisms that underlie the disorder. The previous emphasis on putative peripheral nociceptive mechanisms has advanced to identifying of [...] Read more.

The promulgation of the American College of Rheumatology (ACR) 1990 criteria for fibromyalgia (FM) classification has significantly contributed to an era of increased research into mechanisms that underlie the disorder. The previous emphasis on putative peripheral nociceptive mechanisms has advanced to identifying of changes in central neural networks that modulate pain and other sensory processes. The influences of psychosocial factors on the dynamic and complex neurobiological mechanisms involved in the fibromyalgia clinical phenotype are now better defined. This review highlights key milestones that have directed knowledge concerning the fundamental mechanisms contributing to fibromyalgia. Full article

(This article belongs to the Special Issue Drug Therapies for Fibromyalgia II)

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14 pages, 1059 KiB

Open AccessArticle

Amino Acid Metabolites Associated with Chronic Kidney Disease: An Eight-Year Follow-Up Korean Epidemiology Study

by Hansongyi Lee, Han Byul Jang, Min-Gyu Yoo, Sang Ick Park and Hye-Ja Lee

Biomedicines 2020, 8(7), 222; https://doi.org/10.3390/biomedicines8070222 - 17 Jul 2020

Cited by 48 | Viewed by 4535

Abstract

The discovery of metabolomics-based biomarkers has been a focus of recent kidney dysfunction research. In the present study, we aimed to identify metabolites associated with chronic kidney disease (CKD) in the general population using a cross-sectional study design. At baseline, 6.5% of subjects [...] Read more.

The discovery of metabolomics-based biomarkers has been a focus of recent kidney dysfunction research. In the present study, we aimed to identify metabolites associated with chronic kidney disease (CKD) in the general population using a cross-sectional study design. At baseline, 6.5% of subjects had CKD. Pearson correlation analysis showed that 28 metabolites were significantly associated with estimated glomerular filtration rate (eGFR) after Bonferroni correction. Among these metabolites, 4 acylcarnitines, 12 amino acids, 4 biogenic amines, 1 phosphatidylcholine, and 1 sphingolipid were associated with CKD (p < 0.05). After eight years, 13.5% of subjects had CKD. Three amino acid metabolites were positively associated with new-onset CKD: citrulline [odds ratio (OR): 2.41, 95% confidence interval (CI): 1.26–4.59], kynurenine (OR: 1.98, 95% CI: 1.05–3.73), and phenylalanine (OR: 2.68, 95% CI: 1.00–7.16). The kynurenine:tryptophan ratio was also associated with CKD (OR: 3.20; 95% CI: 1.57–6.51). The addition of multiple metabolites significantly improved the CKD prediction by C statistics (0.756–0.85, p < 0.0001), and the net reclassification improvement was 0.84 (95% CI: 0.72–0.96). Elevated hs-C reactive protein (CRP) was associated with new-onset CKD (OR: 1.045, 95% CI: 1.005–1.086); however, this association disappeared following adjustment with the kynurenine:tryptophan ratio. The levels of citrulline and kynurenine and their ratio to tryptophan in CKD patients with proteinuria were worse than those with one or neither characteristic. Together, the results of this study demonstrate that amino acid metabolites are associated with CKD eight years after initial metabolite assessment. These results could improve the identification of subjects at high risk of CKD who have modified amino acid metabolism. Full article

(This article belongs to the Special Issue Biomarkers in Chronic Kidney Disease)

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12 pages, 635 KiB

Open AccessArticle

Drug-Induced Gingival Overgrowth: The Effect of Cyclosporin A and Mycophenolate Mophetil on Human Gingival Fibroblasts

by Dorina Lauritano, Giulia Moreo, Luisa Limongelli, Annalisa Palmieri and Francesco Carinci

Biomedicines 2020, 8(7), 221; https://doi.org/10.3390/biomedicines8070221 - 17 Jul 2020

Cited by 55 | Viewed by 4072

Abstract

Drug-induced gingival overgrowth may occur after a chronic administration of three classes of systemic drugs: Anticonvulsants, immunosuppressants, and calcium channel blockers. This study aimed to investigate how cyclosporin A and mycophenolate mophetil (immunosuppressive drugs) could interfere with human gingival fibroblasts functions, leading to [...] Read more.

Drug-induced gingival overgrowth may occur after a chronic administration of three classes of systemic drugs: Anticonvulsants, immunosuppressants, and calcium channel blockers. This study aimed to investigate how cyclosporin A and mycophenolate mophetil (immunosuppressive drugs) could interfere with human gingival fibroblasts functions, leading to gingival enlargement. Human gingival fibroblasts derived from the tissue of a 60-year-old female were cultured in a DMEME medium. A stock solution with 1 mg/mL of mycophenolate and 1 mg/mL of cyclosporine were prepared and dissolved in a DMEM medium to prepare a serial dilution at the concentrations of 5000, 2000, 1000, 500, and 100 ng/mL, for both treatments. Cell viability was measured using the PrestoBlue™ Reagent Protocol. Quantitative real-time RT-PCR was performed in order to analyze the expression of 57 genes coding for gingival fibroblasts “Extracellular Matrix and Adhesion Molecules”. Mycophenolate and cyclosporine had no effect on fibroblast cell viability at 1000 ng/mL. Both the treatments showed similar effects on the expression profiling of treated cells: Downregulation of most extracellular matrix metalloproteases genes (MMP8, MMP11, MMP15, MMP16, MMP24) was assessed, while CDH1, ITGA2, ITGA7, LAMB3, MMP12, and MMP13 were recorded to be upregulated in fibroblasts treated with immunosuppressive drugs. It has been demonstrated that gingival overgrowth can be caused by the chronic administration of cyclosporin A and mycophenolate mophetil. However, given the contrasting data of literature, further investigations are needed, making clear the possible effects of immunosuppressive drugs on fibroblasts. Full article

(This article belongs to the Special Issue Models for Oral Biology Research)

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13 pages, 1520 KiB

Open AccessArticle

Inactivated Platelet Lysate Supports the Proliferation and Immunomodulant Characteristics of Mesenchymal Stromal Cells in GMP Culture Conditions

by Katia Mareschi, Sara Castiglia, Aloe Adamini, Deborah Rustichelli, Elena Marini, Alessia Giovanna Santa Banche Niclot, Massimiliano Bergallo, Luciana Labanca, Ivana Ferrero and Franca Fagioli

Biomedicines 2020, 8(7), 220; https://doi.org/10.3390/biomedicines8070220 - 17 Jul 2020

Cited by 7 | Viewed by 2682

Abstract

Mesenchymal stromal cells (MSCs) isolated from bone marrow (BM-MSCs) are considered advanced therapy medicinal products (ATMPs) and need to be produced according to good manufacturing practice (GMP) in their clinical use. Human platelet lysate (HPL) is a good GMP-compliant alternative to animal serum, [...] Read more.

Mesenchymal stromal cells (MSCs) isolated from bone marrow (BM-MSCs) are considered advanced therapy medicinal products (ATMPs) and need to be produced according to good manufacturing practice (GMP) in their clinical use. Human platelet lysate (HPL) is a good GMP-compliant alternative to animal serum, and we have demonstrated that after pathogen inactivation with psoralen, it was safer and more efficient to use psoralen in the production of MSCs following GMP guidelines. In this study, the MSCs cultivated in fetal bovine serum (FBS-MSC) or inactivated HPL (iHPL-MSC) were compared for their immunomodulatory properties. We studied the effects of MSCs on (1) the proliferation of total lymphocytes (Ly) and on naïve T Ly subsets induced to differentiate in Th1 versus Th2 Ly; (2) the immunophenotype of different T-cell subsets; (3) and the cytokine release to verify Th1, Th2, and Th17 polarization. These were analyzed by using an in vitro co-culture system. We observed that iHPL-MSCs showed the same immunomodulatory properties observed in the FBS-MSC co-cultures. Furthermore, a more efficient effect on the increase of naïve T- cells and in the Th1 cytokine release from iHPL was observed. This study confirms that iHPL, used as a medium supplement, may be considered a good alternative to FBS for a GMP-compliant MSC expansion, and also to preserve their immunomodulatory proprieties. Full article

(This article belongs to the Section Model Systems in Research and Development)

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18 pages, 3158 KiB

Open AccessArticle

Thykamine Extracts from Spinach Reduce Acute Inflammation In Vivo and Downregulate Phlogogenic Functions of Human Blood Neutrophils In Vitro

by Vickie Beaupré, Nathalie Boucher and Isabel Desgagné-Penix

Biomedicines 2020, 8(7), 219; https://doi.org/10.3390/biomedicines8070219 - 16 Jul 2020

Cited by 6 | Viewed by 2783

Abstract

The anti-inflammatory and antioxidant role of Thykamine, a botanical extract of thylakoides obtained from spinach leaves, has been investigated in animal and cellular models. The oxidative properties have been proven by inhibiting NO production (>98%) in J774A.1 cells and by protecting a linoelic [...] Read more.

The anti-inflammatory and antioxidant role of Thykamine, a botanical extract of thylakoides obtained from spinach leaves, has been investigated in animal and cellular models. The oxidative properties have been proven by inhibiting NO production (>98%) in J774A.1 cells and by protecting a linoelic acid emulsion subjected to lipid peroxidation caused by AAPH. Thykamine injected intraperitoneally to rats reduced the inflammatory process of (TNBS)-induced colitis and carrageenan-induced paw edema. As neutrophils are the first cells to migrate to inflammatory sites, the influence of Thykamine on the primary neutrophil functions were studied. Thykamine dose-dependent reduced neutrophil chemiotaxis, phagocytosis, and degranulation. No change in the release of LDH by neutrophils on Thykamine was recorded. Thykamine inhibited by 85% the neutrophil production of O₂⁻. A superoxide recovery activity was observed on a zymography demonstrating a SOD-like enzyme on Thykamine extracts. Spontaneous fluorescence provided by carotenoid and chlorophyll pigments (488/675 nm) detected Thykamine on the surface, in the cytoplasm (mainly central where Golgi are present) and weakly in the nucleus of neutrophils. The results argue that SOD and pigments found in Thykamine are part of its antioxidant and anti-inflammatory properties shown in in vivo and in vitro models of inflammation. Full article

(This article belongs to the Section Drug Discovery and Development)

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13 pages, 1570 KiB

Open AccessArticle

Changes in Circulating Extracellular Vesicles in Patients with ST-Elevation Myocardial Infarction and Potential Effects of Remote Ischemic Conditioning—A Randomized Controlled Trial

by Paul M. Haller, Bernhard Jäger, Edita Piackova, Larissa Sztulman, Claudia Wegberger, Johann Wojta, Mariann Gyöngyösi, Attila Kiss, Bruno K. Podesser, Andreas Spittler and Kurt Huber

Biomedicines 2020, 8(7), 218; https://doi.org/10.3390/biomedicines8070218 - 16 Jul 2020

Cited by 14 | Viewed by 3499

Abstract

(1) Background: Extracellular vesicles (EVs) have been recognized as a cellular communication tool with cardioprotective properties; however, it is unknown whether cardioprotection by remote ischemic conditioning (RIC) involves EVs. (2) Methods: We randomized patients with ST-elevation myocardial infarction (STEMI) undergoing primary percutaneous coronary [...] Read more.

(1) Background: Extracellular vesicles (EVs) have been recognized as a cellular communication tool with cardioprotective properties; however, it is unknown whether cardioprotection by remote ischemic conditioning (RIC) involves EVs. (2) Methods: We randomized patients with ST-elevation myocardial infarction (STEMI) undergoing primary percutaneous coronary intervention (PCI) to additionally receive a protocol of RIC or a sham-intervention. Blood was taken before and immediately, 24 h, four days and one month after PCI. Additionally, we investigated EVs from healthy volunteers undergoing RIC. EVs were characterized by a high-sensitive flow cytometer (Beckman Coulter Cytoflex S, Krefeld, Germany). (3) Results: We analyzed 32 patients (16 RIC, 16 control) and five healthy volunteers. We investigated platelet-, endothelial-, leukocyte-, monocyte- and granulocyte-derived EVs and their pro-thrombotic sub-populations expressing superficial phosphatidylserine (PS⁺). We did not observe a significant effect of RIC on the numbers of circulating EVs, although granulocyte-derived EVs were significantly higher in the RIC group. In line, RIC had not impact on EVs in healthy volunteers. Additionally, we observed changes of PS⁺/PEV, EEVs and PS⁺/CD15⁺ EVs irrespective of RIC with time following STEMI. 4) Conclusion: We provide further insights into the course of different circulating EVs during the acute and sub-acute phases of STEMI. With respect to the investigated EV populations, RIC seems to have no effect, with only minor differences found for granulocyte EVs. Full article

(This article belongs to the Special Issue Immunoglobulins in Inflammation)

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13 pages, 2671 KiB

Open AccessArticle

Modulation of Renal Injury by Variable Expression of Myo-Inositol Oxygenase (MIOX) via Perturbation in Metabolic Sensors

by Isha Sharma, Fei Deng and Yashpal S. Kanwar

Biomedicines 2020, 8(7), 217; https://doi.org/10.3390/biomedicines8070217 - 16 Jul 2020

Cited by 3 | Viewed by 3108 | Correction

Abstract

Obesity is associated with perturbations in cellular energy homeostasis and consequential renal injury leading to chronic renal disease (CKD). Myo-inositol oxygenase (MIOX), a tubular enzyme, alters redox balance and subsequent tubular injury in the settings of obesity. Mechanism(s) for such adverse changes remain [...] Read more.

Obesity is associated with perturbations in cellular energy homeostasis and consequential renal injury leading to chronic renal disease (CKD). Myo-inositol oxygenase (MIOX), a tubular enzyme, alters redox balance and subsequent tubular injury in the settings of obesity. Mechanism(s) for such adverse changes remain enigmatic. Conceivably, MIOX accentuates renal injury via reducing expression/activity of metabolic sensors, which perturb mitochondrial dynamics and, if sustained, would ultimately contribute towards CKD. In this brief communication, we utilized MIOX-TG (Transgenic) and MIOX^KO mice, and subjected them to high fat diet (HFD) administration. In addition, ob/ob and ob/MIOX^KO mice of comparable age were used. Mice fed with HFD had increased MIOX expression and remarkable derangements in tubular injury biomarkers. Decreased expression of p-AMPKα (phospho AMP-activated protein kinase) in the tubules was also observed, and it was accentuated in MIOX-TG mice. Interestingly, ob/ob mice also had decreased p-AMPKα expression, which was restored in ob/MIOX^KO mice. Parallel changes were observed in Sirt1/Sirt3 (silent mating type information regulation 2 homolog), and expression of other metabolic sensors, i.e., PGC-1α (Peroxisome proliferator-activated receptor gamma coactivator 1-alpha) and Yin Yang (YY-1). In vitro experiments with tubular cells subjected to palmitate-BSA and MIOX-siRNA had results in conformity with the in vivo observations. These findings link the biology of metabolic sensors to MIOX expression in impaired cellular energy homeostasis with exacerbation/amelioration of renal injury. Full article

(This article belongs to the Special Issue Regenerative Medicine in Diabetes)

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15 pages, 9448 KiB

Open AccessEditor’s ChoiceArticle

Proteome of Glioblastoma-Derived Exosomes as a Source of Biomarkers

by Stanislav Naryzhny, Andrey Volnitskiy, Arthur Kopylov, Elena Zorina, Roman Kamyshinsky, Viktor Bairamukov, Luiza Garaeva, Anatoly Shlikht and Tatiana Shtam

Biomedicines 2020, 8(7), 216; https://doi.org/10.3390/biomedicines8070216 - 16 Jul 2020

Cited by 45 | Viewed by 4454

Abstract

Extracellular vesicles (EV) are involved in important processes of glioblastoma multiforme (GBM), including malignancy and invasion. EV secreted by glioblastoma cells may cross the hematoencephalic barrier and carry molecular cargo derived from the tumor into the peripheral circulation. Therefore, the determination of the [...] Read more.

Extracellular vesicles (EV) are involved in important processes of glioblastoma multiforme (GBM), including malignancy and invasion. EV secreted by glioblastoma cells may cross the hematoencephalic barrier and carry molecular cargo derived from the tumor into the peripheral circulation. Therefore, the determination of the molecular composition of exosomes released by glioblastoma cells seems to be a promising approach for the development of non-invasive methods of the detection of the specific exosomal protein markers in the peripheral blood. The present study aimed to determine the common exosomal proteins presented in preparations from different cell lines and search potential glioblastoma biomarkers in exosomes. We have performed proteomics analysis of exosomes obtained from the conditioned culture medium of five glioblastoma cell lines. A list of 133 proteins common for all these samples was generated. Based on the data obtained, virtual two-dimensional electrophoresis (2DE) maps of proteins presented in exosomes of glioblastoma cells were constructed and the gene ontology (GO) analysis of exosome proteins was performed. A correlation between overexpressed in glial cell proteins and their presence in exosomes have been found. Thus, the existence of many potential glioblastoma biomarkers in exosomes was confirmed. Full article

(This article belongs to the Special Issue The Role of Extracellular Vesicles in Pathogenesis and Therapeutic Strategies)

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10 pages, 1944 KiB

Open AccessArticle

Ectopic Expression of Hematopoietic SHIP1 in Human Colorectal Cancer

by Matthias Schaks, Kristina Allgoewer, Nina Nelson, Patrick Ehm, Asmus Heumann, Florian Ewald, Udo Schumacher, Ronald Simon, Guido Sauter and Manfred Jücker

Biomedicines 2020, 8(7), 215; https://doi.org/10.3390/biomedicines8070215 - 15 Jul 2020

Cited by 3 | Viewed by 2648

Abstract

Colorectal cancer (CRC) is a heterogeneous disease that results from the accumulation of mutations in colonic mucosa cells. A subclass of CRC is characterized by microsatellite instability, which is thought to occur mainly through inactivation of the DNA mismatch repair genes MLH1 and [...] Read more.

Colorectal cancer (CRC) is a heterogeneous disease that results from the accumulation of mutations in colonic mucosa cells. A subclass of CRC is characterized by microsatellite instability, which is thought to occur mainly through inactivation of the DNA mismatch repair genes MLH1 and MSH2. The inositol 5-phosphatase SHIP1 is expressed predominantly in hematopoietic cells. In this study, the expression of SHIP1 in carcinomas and its putative correlation with clinicopathologic parameters, expression of DNA repair genes and microsatellite instability was investigated. By analyzing a multi-tumor tissue microarray, expression of SHIP1 was detected in 48 out of 72 cancer entities analyzed. The expression of SHIP1 protein of 145 kDa was confirmed by Western blot analysis in 7 out of 14 carcinoma cell lines. Analysis of a large colorectal cancer tissue microarray with 1009 specimens revealed SHIP1 expression in 62% of the samples analyzed. SHIP1 expression was inversely correlated with lymph node metastasis, vascular invasion and tumor grade, and it was positively associated with left-sided tumor localization. Interestingly, a strong relationship between the expression of SHIP1 and nuclear and membranous beta-catenin and the DNA repair genes MLH1 and MSH2 was observed. Full article

(This article belongs to the Section Gene and Cell Therapy)

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22 pages, 2735 KiB

Open AccessArticle

Intrinsic Type 1 Interferon (IFN1) Profile of Uncultured Human Bone Marrow CD45^lowCD271⁺ Multipotential Stromal Cells (BM-MSCs): The Impact of Donor Age, Culture Expansion and IFNα and IFNβ Stimulation

by Payal Ganguly, Agata N. Burska, Charlotte L.M. Davis, Jehan J. El-Jawhari, Peter V. Giannoudis and Elena A. Jones

Biomedicines 2020, 8(7), 214; https://doi.org/10.3390/biomedicines8070214 - 15 Jul 2020

Cited by 8 | Viewed by 3652

Abstract

Skeletal aging is associated with reduced proliferative potential of bone marrow (BM) multipotential stromal cells (MSCs). Recent data suggest the involvement of type 1 interferon (IFN1) signalling in hematopoietic stem cell (HSC) senescence. Considering that BM-HSCs and BM-MSCs share the same BM niche, [...] Read more.

Skeletal aging is associated with reduced proliferative potential of bone marrow (BM) multipotential stromal cells (MSCs). Recent data suggest the involvement of type 1 interferon (IFN1) signalling in hematopoietic stem cell (HSC) senescence. Considering that BM-HSCs and BM-MSCs share the same BM niche, we investigated IFN1 expression profile in human BM-MSCs in relation to donor age, culture-expansion and IFN1 (α and β) stimulation. Fluorescence-activated cell sorting was used to purify uncultured BM-MSCs from younger (19–41, n = 6) and older (59–89, n = 6) donors based on the CD45^lowCD271⁺ phenotype, and hematopoietic-lineage cells (BM-HLCs, CD45⁺CD271⁻) were used as controls. Gene expression was analysed using integrated circuits arrays in sorted fractions as well as cultured/stimulated BM-MSCs and Y201/Y202 immortalised cell lines. IFN1 stimulation led to BM-MSC growth arrest and upregulation of many IFN1-stimulated genes (ISGs), with IFNβ demonstrating stronger effects. Uncultured MSCs were characterised by a moderate-level ISG expression similar to Y201 cells. Age-related changes in ISG expression were negligible in BM-MSCs compared to BM-HLCs, and intracellular reactive oxygen species (ROS) levels in BM-MSCs did not significantly correlate with donor age. Antiaging genes Klotho and SIRT6 correlated with more ISGs in BM-MSCs than in BM-HLCs. In patients with osteoarthritis (OA), BM-MSCs expressed considerably lower levels of several ISGs, indicating that their IFN1 signature is affected in a pathological condition. In summary, BM-MSCs possess homeostatic IFN1 gene expression signature in health, which is sensitive to in vitro culture and external IFN1 stimulation. IFN signalling may facilitate in vivo BM-MSC responses to DNA damage and combating senescence and aberrant immune activation. Full article

(This article belongs to the Section Tumor Cell Biology)

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13 pages, 4591 KiB

Open AccessArticle

Fluorescent Labeling of Helminth Extracellular Vesicles Using an In Vivo Whole Organism Approach

by Anders T. Boysen, Bradley Whitehead, Allan Stensballe, Anna Carnerup, Tommy Nylander and Peter Nejsum

Biomedicines 2020, 8(7), 213; https://doi.org/10.3390/biomedicines8070213 - 14 Jul 2020

Cited by 21 | Viewed by 4874

Abstract

In the last two decades, extracellular vesicles (EVs) from the three domains of life, Archaea, Bacteria and Eukaryotes, have gained increasing scientific attention. As such, the role of EVs in host-pathogen communication and immune modulation are being intensely investigated. Pivotal to EV research [...] Read more.

In the last two decades, extracellular vesicles (EVs) from the three domains of life, Archaea, Bacteria and Eukaryotes, have gained increasing scientific attention. As such, the role of EVs in host-pathogen communication and immune modulation are being intensely investigated. Pivotal to EV research is the determination of how and where EVs are taken up by recipient cells and organs in vivo, which requires suitable tracking strategies including labelling. Labelling of EVs is often performed post-isolation which increases risks of non-specific labelling and the introduction of labelling artefacts. Here we exploited the inability of helminths to de novo synthesise fatty acids to enable labelling of EVs by whole organism uptake of fluorescent lipid analogues and the subsequent incorporation in EVs. We showed uptake of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (DOPE-Rho) in Anisakis spp. and Trichuris suis larvae. EVs isolated from the supernatant of Anisakis spp. labelled with DOPE-Rho were characterised to assess the effects of labelling on size, structure and fluorescence of EVs. Fluorescent EVs were successfully taken up by the human macrophage cell line THP-1. This study, therefore, presents a novel staining method that can be utilized by the EV field in parasitology and potentially across multiple species. Full article

(This article belongs to the Special Issue The Role of Extracellular Vesicles in Pathogenesis and Therapeutic Strategies)

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16 pages, 1551 KiB

Open AccessArticle

Reactive Metamizole Metabolites Enhance the Toxicity of Hemin on the ATP Pool in HL60 Cells by Inhibition of Glycolysis

by Deborah Rudin, Maurice Schmutz, Noëmi Johanna Roos, Jamal Bouitbir and Stephan Krähenbühl

Biomedicines 2020, 8(7), 212; https://doi.org/10.3390/biomedicines8070212 - 14 Jul 2020

Cited by 3 | Viewed by 3214

Abstract

Metamizole is an analgesic, whose pharmacological and toxicological properties are attributed to N-methyl-aminoantipyrine (MAA), its major metabolite. In the presence of heme iron, MAA forms reactive metabolites, which are toxic for granulocyte precursors. Since decreased cellular ATP is characteristic for MAA-associated toxicity, we [...] Read more.

Metamizole is an analgesic, whose pharmacological and toxicological properties are attributed to N-methyl-aminoantipyrine (MAA), its major metabolite. In the presence of heme iron, MAA forms reactive metabolites, which are toxic for granulocyte precursors. Since decreased cellular ATP is characteristic for MAA-associated toxicity, we studied the effect of MAA with and without hemin on energy metabolism of HL60 cells, a granulocyte precursor cell line. The combination MAA/hemin depleted the cellular ATP stronger than hemin alone, whereas MAA alone was not toxic. This decrease in cellular ATP was observed before plasma membrane integrity impairment. MAA/hemin and hemin did not affect the proton leak but increased the maximal oxygen consumption by HL60 cells. This effect was reversed by addition of the radical scavenger N-acetylcysteine. The mitochondrial copy number was not affected by MAA/hemin or hemin. Hemin increased mitochondrial superoxide generation, which was not accentuated by MAA. MAA decreased cellular ROS accumulation in the presence of hemin. In cells cultured in galactose (favoring mitochondrial ATP generation), MAA/hemin had less effect on the cellular ATP and plasma membrane integrity than in glucose. MAA/hemin impaired glycolysis more than hemin or MAA alone, and N-acetylcysteine blunted this effect of MAA/hemin. MAA/hemin decreased protein expression of pyruvate kinase more than hemin or MAA alone. In conclusion, cellular ATP depletion appears to be an important mechanism of MAA/hemin toxicity on HL60 cells. MAA itself is not toxic on HL60 cells up to 100 µM but boosts the inhibitory effect of hemin on glycolysis through the formation of reactive metabolites. Full article

(This article belongs to the Section Tumor Cell Biology)

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15 pages, 1905 KiB

Open AccessArticle

Specific and Non-Invasive Fluorescent Labelling of Extracellular Vesicles for Evaluation of Intracellular Processing by Intestinal Epithelial Cells

by Maria S. Hansen, Ida S. E. Gadegaard, Eva C. Arnspang, Kristine Blans, Lene N. Nejsum and Jan T. Rasmussen

Biomedicines 2020, 8(7), 211; https://doi.org/10.3390/biomedicines8070211 - 14 Jul 2020

Cited by 16 | Viewed by 4717

Abstract

The presence of extracellular vesicles (EVs) in milk has gained interest due to their capacity to modulate the infant’s intestinal and immune system. Studies suggest that milk EVs are enriched in immune-modulating proteins and miRNA, highlighting their possible health benefits to infants. To [...] Read more.

The presence of extracellular vesicles (EVs) in milk has gained interest due to their capacity to modulate the infant’s intestinal and immune system. Studies suggest that milk EVs are enriched in immune-modulating proteins and miRNA, highlighting their possible health benefits to infants. To assess uptake of milk EVs by intestinal epithelial cells, a method was developed using labelling of isolated EVs with fluorophore-conjugated lactadherin. Lactadherin is a generic and validated EV marker, which enables an effective labelling of phosphatidylserine (PS) exposing EVs. Labelled EVs could effectively be used to describe a dose- and time-dependent uptake into the intestinal epithelial Caco-2 cell line. Additionally, fluorescence microscopy was employed to show that EVs colocalize with endosomal markers and lysosomes, indicating that EVs are taken up via general endocytotic mechanisms. Collectively, a method to specifically label isolated EVs is presented and employed to study the uptake of milk EVs by intestinal epithelial cells. Full article

(This article belongs to the Special Issue The Role of Extracellular Vesicles in Pathogenesis and Therapeutic Strategies)

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18 pages, 842 KiB

Open AccessReview

Endoplasmic Reticulum Stress in Macrophages: The Vicious Circle of Lipid Accumulation and Pro-Inflammatory Response

by Vasily N. Sukhorukov, Victoria A. Khotina, Mariam Bagheri Ekta, Ekaterina A. Ivanova, Igor A. Sobenin and Alexander N. Orekhov

Biomedicines 2020, 8(7), 210; https://doi.org/10.3390/biomedicines8070210 - 13 Jul 2020

Cited by 29 | Viewed by 5861

Abstract

The endoplasmic reticulum (ER) stress is an important event in the pathogenesis of different human disorders, including atherosclerosis. ER stress leads to disturbance of cellular homeostasis, apoptosis, and in the case of macrophages, to foam cell formation and pro-inflammatory cytokines production. In atherosclerosis, [...] Read more.

The endoplasmic reticulum (ER) stress is an important event in the pathogenesis of different human disorders, including atherosclerosis. ER stress leads to disturbance of cellular homeostasis, apoptosis, and in the case of macrophages, to foam cell formation and pro-inflammatory cytokines production. In atherosclerosis, several cell types can be affected by ER stress, including endothelial cells, vascular smooth muscular cells, and macrophages. Modified low-density lipoproteins (LDL) and cytokines, in turn, can provoke ER stress through different processes. The signaling cascades involved in ER stress initiation are complex and linked to other cellular processes, such as lysosomal biogenesis and functioning, autophagy, mitochondrial homeostasis, and energy production. In this review, we discuss the underlying mechanisms of ER stress formation and the interplay of lipid accumulation and pro-inflammatory response. We will specifically focus on macrophages, which are the key players in maintaining chronic inflammatory milieu in atherosclerotic lesions, and also a major source of lipid-accumulating foam cells. Full article

(This article belongs to the Special Issue Macrophages in Health and Non-infectious Disease)

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13 pages, 2427 KiB

Open AccessArticle

Aggregate Removal Nanofiltration of Human Serum Albumin Solution Using Nanocellulose-Based Filter Paper

by Lulu Wu, Athanasios Mantas, Simon Gustafsson, Levon Manukyan and Albert Mihranyan

Biomedicines 2020, 8(7), 209; https://doi.org/10.3390/biomedicines8070209 - 13 Jul 2020

Cited by 8 | Viewed by 4434

Abstract

This study is dedicated to the rapid removal of protein aggregates and viruses from plasma-derived human serum albumin (HSA) product to reduce the risk of viral contamination and increase biosafety. A two-step filtration approach was implemented to first remove HSA aggregates and then [...] Read more.

This study is dedicated to the rapid removal of protein aggregates and viruses from plasma-derived human serum albumin (HSA) product to reduce the risk of viral contamination and increase biosafety. A two-step filtration approach was implemented to first remove HSA aggregates and then achieve high model virus clearance using a nanocellulose-based filter paper of different thicknesses, i.e., 11 μm (prefilter) and 22 μm (virus filter) at pH 7.4 and room temperature. The pore size distribution of these filters was characterized by nitrogen gas sorption analysis. Dynamic light scattering (DLS) and size-exclusion high performance liquid chromatography (SE-HPLC) were performed to analyze the presence of HSA aggregates in process intermediates. The virus filter showed high clearance of a small-size model virus, i.e., log₁₀ reduction value (LRV) > 5, when operated at 3 and 5 bar, but a distinct decrease in LRV was detected at 1 bar, i.e., LRV 2.65–3.75. The throughput of HSA was also dependent on applied transmembrane pressure as was seen by Vmax values of 110 ± 2.5 L m⁻² and 63.6 ± 5.8 L m⁻² at 3 bar and 5 bar, respectively. Protein loss was low, i.e., recovery > 90%. A distribution of pore sizes between 40 nm and 60 nm, which was present in the prefilter and absent in the virus filter, played a crucial part in removing the HSA aggregates and minimizing the risk of virus filter fouling. The presented results enable the application of virus removal nanofiltration of HSA in bioprocessing as an alternative to virus inactivation methods based, e.g., on heat treatment. Full article

(This article belongs to the Section Biomedical Materials and Nanomedicine)

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25 pages, 7688 KiB

Open AccessReview

Potential Endogenous Cell Sources for Retinal Regeneration in Vertebrates and Humans: Progenitor Traits and Specialization

by Eleonora N. Grigoryan

Biomedicines 2020, 8(7), 208; https://doi.org/10.3390/biomedicines8070208 - 12 Jul 2020

Cited by 10 | Viewed by 3963

Abstract

Retinal diseases often cause the loss of photoreceptor cells and, consequently, impairment of vision. To date, several cell populations are known as potential endogenous retinal regeneration cell sources (RRCSs): the eye ciliary zone, the retinal pigment epithelium, the iris, and Müller glia. Factors [...] Read more.

Retinal diseases often cause the loss of photoreceptor cells and, consequently, impairment of vision. To date, several cell populations are known as potential endogenous retinal regeneration cell sources (RRCSs): the eye ciliary zone, the retinal pigment epithelium, the iris, and Müller glia. Factors that can activate the regenerative responses of RRCSs are currently under investigation. The present review considers accumulated data on the relationship between the progenitor properties of RRCSs and the features determining their differentiation. Specialized RRCSs (all except the ciliary zone in low vertebrates), despite their differences, appear to be partially “prepared” to exhibit their plasticity and be reprogrammed into retinal neurons due to the specific gene expression and epigenetic landscape. The “developmental” characteristics of RRCS gene expression are predefined by the pathway by which these cell populations form during eye morphogenesis; the epigenetic features responsible for chromatin organization in RRCSs are under intracellular regulation. Such genetic and epigenetic readiness is manifested in vivo in lower vertebrates and in vitro in higher ones under conditions permissive for cell phenotype transformation. Current studies on gene expression in RRCSs and changes in their epigenetic landscape help find experimental approaches to replacing dead cells through recruiting cells from endogenous resources in vertebrates and humans. Full article

(This article belongs to the Section Model Systems in Research and Development)

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9 pages, 430 KiB

Open AccessArticle

Serum γ-Glutamyltransferase Concentration Predicts Endothelial Dysfunction in Naïve Hypertensive Patients

by Maria Perticone, Raffaele Maio, Benedetto Caroleo, Angela Sciacqua, Edoardo Suraci, Simona Gigliotti, Francesco Martino, Francesco Andreozzi, Giorgio Sesti and Francesco Perticone

Biomedicines 2020, 8(7), 207; https://doi.org/10.3390/biomedicines8070207 - 11 Jul 2020

Cited by 3 | Viewed by 2337

Abstract

Background: Serum gamma-glutamyltransferase (γ-GT) is recognized as a risk factor for cardiovascular diseases (CV). Traditional cardiovascular risk factors mediate endothelial dysfunction. Aim: to evaluate a possible correlation between serum γ-GT and endothelium-dependent vasodilation in naïve hypertensives. Methods: We enrolled 500 hypertensives. Endothelial function [...] Read more.

Background: Serum gamma-glutamyltransferase (γ-GT) is recognized as a risk factor for cardiovascular diseases (CV). Traditional cardiovascular risk factors mediate endothelial dysfunction. Aim: to evaluate a possible correlation between serum γ-GT and endothelium-dependent vasodilation in naïve hypertensives. Methods: We enrolled 500 hypertensives. Endothelial function was studied by strain-gauge plethysmography. Receiver operating characteristic (ROC) analysis was used to assess the predictive value of γ-GT and to identify the optimal cut-off value of the same variable for endothelial dysfunction. Results: At univariate linear analysis peak percent increase in acetylcholine (ACh)-stimulated vasodilation was inversely related to γ-GT (r = −0.587), alanine aminotransferase (ALT) (r = −0.559), aspartate aminotransferase (AST) (r = −0.464), age (r = −0.171), body mass index (BMI) (r = −0.152), and fasting glucose (r = −101). In the stepwise multivariate regression model, endothelium-dependent vasodilation was significantly related to γ-GT (β = −0.362), ALT (β = −0.297), AST (β = −0.217), estimated glomerular filtration rate (e-GFR) (β = 0.199), gender (β = 0.166), and smoking (β = −0.061). The ROC analysis demonstrated that the accuracy of γ-GT for identifying patients with endothelial dysfunction was 82.1%; the optimal γ-GT cut-off value for discriminating patients with this alteration was 27 UI/L. Conclusions: Serum γ-GT values, within the normal range, are significantly associated with endothelial dysfunction in hypertensives, and may be considered a biomarker of early vascular damage. Full article

(This article belongs to the Special Issue Endothelial Dysfunction: From Pathophysiology to Novel Therapeutic Approaches)

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12 pages, 1315 KiB

Open AccessReview

NADPH Oxidases and Their Role in Atherosclerosis

by Anastasia V. Poznyak, Andrey V. Grechko, Varvara A. Orekhova, Victoria Khotina, Ekaterina A. Ivanova and Alexander N. Orekhov

Biomedicines 2020, 8(7), 206; https://doi.org/10.3390/biomedicines8070206 - 10 Jul 2020

Cited by 59 | Viewed by 5137

Abstract

The current view on atherosclerosis positions it as a multifactorial disorder that results from the interplay between lipid metabolism disturbances and inflammatory processes. Oxidative stress is proven to be one of the initiating factors in atherosclerosis development, being implicated both in the inflammatory [...] Read more.

The current view on atherosclerosis positions it as a multifactorial disorder that results from the interplay between lipid metabolism disturbances and inflammatory processes. Oxidative stress is proven to be one of the initiating factors in atherosclerosis development, being implicated both in the inflammatory response and in atherogenic modifications of lipoproteins that facilitate lipid accumulation in the arterial wall. The hallmark of oxidative stress is the elevated level of reactive oxygen species (ROS). Correspondingly, the activity of major ROS-generating enzymes, including nicotinamide adenine dinucleotide phosphate (NADPH) oxidases, xanthine oxidases, and cyclooxygenases, is an important element in atherosclerosis development. In particular, the role of NADPH oxidases in atherosclerosis development has become a subject of intensive research. Aberrant activity of NADPH oxidases was shown to be associated with cardiovascular disease in humans. With regard to atherosclerosis, several important pathological components of the disease development, including endothelial dysfunction, inflammation, and vascular remodeling, involve aberrations in NADPH oxidases functioning. In humans, NADPH oxidases are represented by four isoforms expressed in vascular tissues, where they serve as the main source of ROS during atherogenesis. Moreover, recent studies have demonstrated their impact on vascular remodeling processes. Interestingly, one of the NADPH oxidase isoforms, NOX4, was shown to have an atheroprotective effect. Despite the growing evidence of the crucial involvement of NADPH oxidases in atherosclerosis pathogenesis, the available data still remains controversial. In this narrative review, we summarize the current knowledge of the role of NADPH oxidases in atherosclerosis and outline the future directions of research. Full article

(This article belongs to the Section Molecular and Translational Medicine)

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11 pages, 694 KiB

Open AccessReview

NLPR3 Inflammasomes and Their Significance for Atherosclerosis

by Anastasia V. Poznyak, Alexandra A. Melnichenko, Reinhard Wetzker, Elena V. Gerasimova and Alexander N. Orekhov

Biomedicines 2020, 8(7), 205; https://doi.org/10.3390/biomedicines8070205 - 10 Jul 2020

Cited by 34 | Viewed by 4441

Abstract

Atherosclerosis is a serious disorder, with numerous potential complications such as cardiovascular disease, ischemic stroke, and myocardial infarction. The origin of atherosclerosis is related to chronic inflammation, lipid metabolism alterations, and oxidative stress. Inflammasomes are the cytoplasmic multiprotein complex triggering the activation of [...] Read more.

Atherosclerosis is a serious disorder, with numerous potential complications such as cardiovascular disease, ischemic stroke, and myocardial infarction. The origin of atherosclerosis is related to chronic inflammation, lipid metabolism alterations, and oxidative stress. Inflammasomes are the cytoplasmic multiprotein complex triggering the activation of inflammatory response. NLRP3 inflammasomes have a specific activation pathway that involves numerous stimuli, including a wide range of PAMPs and DAMPs. Recent studies of atherosclerotic pathology are focused on the mitochondria that appear to be a promising target for therapeutic approach development. Mitochondria are the main source of reactive oxygen species (ROS) associated with oxidative stress. It was previously shown that NLRP3 inflammasome activation results in mitochondrial damage, but the exact mechanisms of this need to be specified. In this review, we focused on the features of NLRP3 inflammasomes and their significance for atherosclerosis, especially concerning mitochondria. Full article

(This article belongs to the Section Molecular and Translational Medicine)

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16 pages, 894 KiB

Open AccessEditor’s ChoiceReview

Bi- and Tri-Specific T Cell Engager-Armed Oncolytic Viruses: Next-Generation Cancer Immunotherapy

by Zong Sheng Guo, Michael T. Lotze, Zhi Zhu, Walter J. Storkus and Xiao-Tong Song

Biomedicines 2020, 8(7), 204; https://doi.org/10.3390/biomedicines8070204 - 10 Jul 2020

Cited by 48 | Viewed by 8453

Abstract

Oncolytic viruses (OVs) are potent anti-cancer biologics with a bright future, having substantial evidence of efficacy in patients with cancer. Bi- and tri-specific antibodies targeting tumor antigens and capable of activating T cell receptor signaling have also shown great promise in cancer immunotherapy. [...] Read more.

Oncolytic viruses (OVs) are potent anti-cancer biologics with a bright future, having substantial evidence of efficacy in patients with cancer. Bi- and tri-specific antibodies targeting tumor antigens and capable of activating T cell receptor signaling have also shown great promise in cancer immunotherapy. In a cutting-edge strategy, investigators have incorporated the two independent anti-cancer modalities, transforming them into bi- or tri-specific T cell engager (BiTE or TriTE)-armed OVs for targeted immunotherapy. Since 2014, multiple research teams have studied this combinatorial strategy, and it showed substantial efficacy in various tumor models. Here, we first provide a brief overview of the current status of oncolytic virotherapy and the use of multi-specific antibodies for cancer immunotherapy. We then summarize progress on BiTE and TriTE antibodies as a novel class of cancer therapeutics in preclinical and clinical studies, followed by a discussion of BiTE- or TriTE-armed OVs for cancer therapy in translational models. In addition, T cell receptor mimics (TCRm) have been developed into BiTEs and are expected to greatly expand the application of BiTEs and BiTE-armed OVs for the effective targeting of intracellular tumor antigens. Future applications of such innovative combination strategies are emerging as precision cancer immunotherapies. Full article

(This article belongs to the Special Issue Oncolytic Viruses as a Novel Form of Immunotherapy for Cancer II)

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15 pages, 1771 KiB

Open AccessArticle

Trained Immunity Based-Vaccines as a Prophylactic Strategy in Common Variable Immunodeficiency. A Proof of Concept Study

by Kissy Guevara-Hoyer, Paula Saz-Leal, Carmen M. Diez-Rivero, Juliana Ochoa-Grullón, Miguel Fernández-Arquero, Rebeca Pérez de Diego and Silvia Sánchez-Ramón

Biomedicines 2020, 8(7), 203; https://doi.org/10.3390/biomedicines8070203 - 9 Jul 2020

Cited by 20 | Viewed by 4584

Abstract

Background. A major concern in the care of common variable immunodeficiency (CVID) patients is the persistence of subclinical or recurrent respiratory tract infections (RRTI) despite adequate trough IgG levels, which impacts the quality of life (QoL) and morbidity. Therefore, the development of new [...] Read more.

Background. A major concern in the care of common variable immunodeficiency (CVID) patients is the persistence of subclinical or recurrent respiratory tract infections (RRTI) despite adequate trough IgG levels, which impacts the quality of life (QoL) and morbidity. Therefore, the development of new approaches to prevent and treat infection, especially RRTI, is necessary. Objectives. We conducted a clinical observational study from May, 2016 to December, 2017 in 20 CVID patients; ten of these patients had a history of RRTI and received the polybacterial preparation MV130, a trained immunity-based vaccine (TIbV) to assess its impact on their QoL and prognosis. Methods. Subjects with RRTI received MV130 for 3 months and were followed up to 12 months after initiation of the treatment. The primary endpoint was a reduction in RRTI at the end of the study. We analyzed the pharmacoeconomic impact on the RRTI group before and after immunotherapy by estimating the direct and indirect costs, and assessed CVID-QoL and cytokine profile. Specific antibody responses to the bacteria contained in MV130 were measured. Results. The RRTI-group treated with TIbV MV130 showed a significant decrease in infection rate (p = 0.006) throughout the 12 months after initiation of the treatment. A decrease in antibiotic use and unscheduled outpatient visits was observed (p = 0.005 and p = 0.002, respectively). Significant increases in anti-pneumococcus and anti-MV130 IgA antibodies (p = 0.039 both) were detected after 12 months of MV130. Regarding the CVID QoL questionnaire, an overall decrease in the score by more than 50% was observed (p < 0.05) which demonstrated that patients experienced an improvement in their QoL. The pharmacoeconomic analysis showed that the real annual direct costs decreased up to 4 times per patient with the prophylactic intervention (p = 0.005). Conclusion. The sublingual administration of the TIbV MV130 significantly reduced the rate of respiratory infections, antibiotic use and unscheduled visits, while increasing specific IgA responses in CVID patients. Additionally, the CVID population felt that their QoL was improved, and a decrease in expenses derived from health care was predicted. Full article

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Biomedicines, Volume 8, Issue 7 (July 2020) – 54 articles

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