Molecular Responses in THP-1 Macrophage-Like Cells Exposed to Diverse Nanoparticles

In the body, engineered nanoparticles (NPs) may be recognized and processed by immune cells, among which macrophages play a crucial role. We evaluated the effects of selected NPs [NM-100 (TiO2), NM-110 (ZnO), NM-200 (SiO2), and NM-300 K (Ag)] on THP-1 macrophage-like cells. The cells were exposed to subcytotoxic concentrations of NPs (1–25 µg/mL) and the expression of immunologically relevant genes (VCAM1, TNFA, CXCL8, ICAM1, CD86, CD192, and IL1B) was analyzed by RT-qPCR. The expression of selected cytokines, growth factors and surface molecules was assessed by flow cytometry or ELISA. Generation of reactive oxygen species and induction of DNA breaks were also analyzed. Exposure to diverse NPs caused substantially different molecular responses. No significant effects were detected for NM-100 treatment. NM-200 induced production of IL-8, a potent attractor and activator of neutrophils, growth factors (VEGF and IGF-1) and superoxide. NM-110 triggered a proinflammatory response, characterized by the activation of transcription factor NF-κB, an enhanced production of proinflammatory cytokines (TNF-α) and chemokines (IL-8). Furthermore, the expression of cell adhesion molecules VCAM-1 and ICAM-1 and hepatocyte growth factor (HGF), as well as superoxide production and DNA breaks, were affected. NM-300 K enhanced IL-8 production and induced DNA breaks, however, it decreased the expression of chemokine receptor (CCR2) and CD86 molecule, indicating potential immunosuppressive activity. The toxicity of ZnO and Ag NPs was probably caused by their intracellular dissolution, as indicated by transmission electron microscopy imaging. The observed effects in macrophages might further influence both innate and adaptive immune responses by promoting neutrophil recruitment via IL-8 release and enhancing the adhesion and stimulation of T cells by VCAM-1 and ICAM-1 expression.

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Hypothetical Role of Growth Factors to Reduce Intervertebral Disc Degeneration Significantly through Trained Biological Transformations

Current Pharmaceutical Design ◽

10.2174/1381612826666201019104201 ◽

2020 ◽

Vol 26 ◽

Author(s):

Cristian Muresanu ◽

Siva G. Somasundaram ◽

Sergey V. Vissarionov ◽

Liliya V. Gavryushova ◽

Vladimir N. Nikolenko ◽

...

Keyword(s):

Growth Factors ◽

Blood Circulation ◽

Seminal Fluid ◽

Prostate Gland ◽

The Body ◽

Personal Experiences ◽

Biological Transformation ◽

The Mind

Background: From the evidence of failed injection-based growth factor therapies, it has been proposed that a naturally triggered uninterrupted blood circulation of the growth factors would be superior. Objective: We seek to stimulate discussions and more research about the possibility of using the already available growth factors found in the prostate gland and endometrium by starting a novel educable physiology, known as biological transformations controlled by the mind. Methods: We summarized the stretch-gated ion channel mechanism of the cell membrane, and offer several practical methods that can be applied by anyone, in order to stimulate and enhance the blood circulation of the growth factors from the seminal fluid to sites throughout the body. This details the practical application of our earlier published studies about biological transformations. Results: A previously reported single-patient case study has been extended, adding more from his personal experiences continually improving this novel physiological training and extending the ideas from our earlier findings in detail. Conclusion: The biological transformation findings demonstrate the need additional research to establish the benefits of these natural therapies to repair and rejuvenate tissues affected by various chronic diseases or aging processes.

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Structural Insight into CVB3-VLP Non-Adjuvanted Vaccine

Microorganisms ◽

10.3390/microorganisms8091287 ◽

2020 ◽

Vol 8 (9) ◽

pp. 1287

Author(s):

Minna M. Hankaniemi ◽

Mo A. Baikoghli ◽

Virginia M. Stone ◽

Li Xing ◽

Outi Väätäinen ◽

...

Keyword(s):

Exchange Chromatography ◽

Scale Production ◽

Igg Antibody ◽

Purification Method ◽

Microscopy Imaging ◽

Single Particle Reconstruction ◽

Enhanced Production ◽

Chronic Myocarditis ◽

Flow Filtration ◽

Antibody Levels

Coxsackievirus B (CVB) enteroviruses are common pathogens that can cause acute and chronic myocarditis, dilated cardiomyopathy, aseptic meningitis, and they are hypothesized to be a causal factor in type 1 diabetes. The licensed enterovirus vaccines and those currently in clinical development are traditional inactivated or live attenuated vaccines. Even though these vaccines work well in the prevention of enterovirus diseases, new vaccine technologies, like virus-like particles (VLPs), can offer important advantages in the manufacturing and epitope engineering. We have previously produced VLPs for CVB3 and CVB1 in insect cells. Here, we describe the production of CVB3-VLPs with enhanced production yield and purity using an improved purification method consisting of tangential flow filtration and ion exchange chromatography, which is compatible with industrial scale production. We also resolved the CVB3-VLP structure by Cryo-Electron Microscopy imaging and single particle reconstruction. The VLP diameter is 30.9 nm on average, and it is similar to Coxsackievirus A VLPs and the expanded enterovirus cell-entry intermediate (the 135s particle), which is ~2 nm larger than the mature virion. High neutralizing and total IgG antibody levels, the latter being a predominantly Th2 type (IgG1) phenotype, were detected in C57BL/6J mice immunized with non-adjuvanted CVB3-VLP vaccine. The structural and immunogenic data presented here indicate the potential of this improved methodology to produce highly immunogenic enterovirus VLP-vaccines in the future.

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Physiopathology of Spine Metastasis

International Journal of Surgical Oncology ◽

10.1155/2011/107969 ◽

2011 ◽

Vol 2011 ◽

pp. 1-8 ◽

Cited By ~ 25

Author(s):

Giulio Maccauro ◽

Maria Silvia Spinelli ◽

Sigismondo Mauro ◽

Carlo Perisano ◽

Calogero Graci ◽

...

Keyword(s):

Growth Factors ◽

Bone Metastasis ◽

Lymphatic System ◽

Spinal Metastasis ◽

Bone Destruction ◽

The Body ◽

Vicious Cycle ◽

Patients With Cancer ◽

Local Tumour ◽

Neurological Injuries

The metastasis is the spread of cancer from one part of the body to another. Two-thirds of patients with cancer will develop bone metastasis. Breast, prostate and lung cancer are responsible for more than 80% of cases of metastatic bone disease. The spine is the most common site of bone metastasis. A spinal metastasis may cause pain, instability and neurological injuries. The diffusion through Batson venous system is the principal process of spinal metastasis, but the dissemination is possible also through arterial and lymphatic system or by contiguity. Once cancer cells have invaded the bone, they produce growth factors that stimulate osteoblastic or osteolytic activity resulting in bone remodeling with release of other growth factors that lead to a vicious cycle of bone destruction and growth of local tumour.

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Toxicity study of new metal nanoparticles functionalized with fluorescein derivatives as novel image systems

Microscopy and Microanalysis ◽

10.1017/s1431927613000743 ◽

2013 ◽

Vol 19 (S4) ◽

pp. 25-26 ◽

Cited By ~ 2

Author(s):

A. Fernandéz-Lodeiro ◽

J. Fernandéz-Lodeiro ◽

C. Nuñez ◽

E. Oliveira ◽

H.M. Santos ◽

...

Keyword(s):

The Body ◽

Limited Information ◽

Glutathione S Transferase ◽

Ag Nps ◽

One Pot ◽

Novel Treatments ◽

Transmission Electron ◽

Dying Cells ◽

Gastro Intestinal Tract ◽

In Cells

Nanoparticles in general (NPs) and/or nanomaterials offer remarkable opportunities in industrial production, daily consumables, medicine, biotechnology, electronics and numerous other important commercial and economical areas. Among all these areas, nanomedicine has opened novel treatments for problematic diseases such as viral, genetic, cancer, AIDS, etc. There is limited information available regarding translocation and distribution of NPs in the body and in the environment. Additionally, there is also need for more information on NPs toxicity. Recently has been demonstrated that physiological barriers such as pulmonary and gastro-intestinal tract are affected.The main objective of this work is to use functionalized metal NPs, as emissive agent markers, assess their internalization in cells and evaluate toxicity to cells.Using the emissive two probes synthesized in a one-pot reaction using fluoresceine as chromophore, several gold (Au), round shape, and silver (Ag) NPs (round and triangular shapes) were functionalized in organic media and water by Brust and Turkevish methodology, using tetraoctylammonium bromide (TOABr) as a common stabilizer and sodium borohydride as reducing agent. All has been characterized by UV-vis and emission spectroscopy, transmission electron microscopy (TEM) (Figure 1), and Light scattering. To study the route of internalization into the cell NP-complexes were injected intraperitoneally in fish (Carassius auratus). After 48 hour fish were sampled and sacrificed and liver and intestine processed for histology examination. Additional sub-samples were stored at – 80ºC for enzymatic analysis (glutathione-S-transferase and catalase). Blood was also collected from healthy non-injected fish, for leucocyte separation followed by incubation with the metal NPs and cell viability assays. The presence of emissive NPs in cells was examined by microscopy using a Leica microscope (ATC 2000) adapted for epifluorescence (EF).The microscopy analysis showed that apparently both metal NPs were internalized by leucocytes and intestine cells (Figure 2a and 2b) but apparently not by hepatocytes. However, it is still to clarify if NPs internalization occurred in dead or dying cells only, with more permeable membranes, or also in living cells. Another possibility relates to the detection limits and resolution of the microscope used: the fraction of NPs entering is too low and not detectable with this type of equipment. No significant fluorescence was detected in controls. Viability assays showed higher mortality rates in leucocytes incubated with triangular Ag NPs suggesting that the type of metal and shape have influence in cell toxicity. In general, enzymatic assays indicate low oxidative stress for cells. However, GST results show significant (p > 0.05) differences in livers from fish injected with round Ag NPs. With respect to catalase, significant differences (p > 0.05) were detected in livers from fish injected with round Au NPs. Although the presented results are preliminary they suggest that functionalized NPs are able to penetrate cell membranes. On the other hand, the observed toxicity can be attributed to differences in shape and type of metal NPs.The authors acknowledge the funding by Fundação para a Ciência e Tecnologia through grant PTDC/MAR/119068/2010 and through project no. PEst-C/EQB/LA0006/2011 granted to Requimte.

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Cellular and molecular responses of the uterus to embryo implantation can be elicited by locally applied growth factors

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.98.3.1047 ◽

2001 ◽

Vol 98 (3) ◽

pp. 1047-1052 ◽

Cited By ~ 220

Author(s):

B. C. Paria ◽

W.-g. Ma ◽

J. Tan ◽

S. Raja ◽

S. K. Das ◽

...

Keyword(s):

Growth Factors ◽

Embryo Implantation ◽

Molecular Responses

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Retrospective Study of Trough Imatinib Plasma Levels Conducted as Part of the Follow-up of Patients with Chronic Myeloid Leukemia in a Canadian Cohort

Blood ◽

10.1182/blood.v112.11.4258.4258 ◽

2008 ◽

Vol 112 (11) ◽

pp. 4258-4258

Author(s):

Rahima Jamal ◽

Danielle Desmarais ◽

John Chapdelaine ◽

Yvan Côté ◽

Lambert Busque

Keyword(s):

Chronic Myeloid Leukemia ◽

Plasma Levels ◽

Myeloid Leukemia ◽

Plasma Concentrations ◽

The Body ◽

Inadequate Response ◽

Molecular Responses ◽

The Impact ◽

Trough Levels

Abstract While imatinib biodisponibility is excellent, trough imatinib plasma levels associated with standard dose imatinib are variable and cannot be predicted by the age, the body surface area or the weight of the patient. Imatinib trough levels have recently been associated with both cytogenetic and molecular responses, making imatinib pharmacokinetics a possible target in optimisation of the treatment of patients with chronic myeloid leukemia. We retrospectively analysed trough imatinib plasma levels prescribed as part of the longitudinal follow-up of a cohort of patients with chronic myeloid leukemia in Canada. Indications for testing were inadequate response, important side effects or suspicion of non compliance. The objectives of the study were to evaluate the variability of trough imatinib plasma levels in our cohort and determine the impact a first result had on the subsequent plasma level in patients with more than one imatinib plasma determination. Analyses of trough plasma levels in 278 patients were conducted in a central canadian laboratory from April 2007 to April 2008. Trough imatinib plasma levels were measured using liquid chromatography and tandem mass spectrometry (LC/MS/MS) with deuterated imatinib as the internal standard. Distribution of trough imatinib plasma levels according to the established IRIS quartiles (Q1–Q4; BLOOD. 2008, vol 11, p4022)) showed an important variability, with plasma levels distributed between less than 100 ng/ml and more than 4500 ng/ml. Sixty-two (22.3%) patients in our cohort had plasma levels below 647 ng/ml (Q1), 101(36.3%) patients had levels between 647–1170 ng/ml (Q2–Q3) and 115 (41.3%) patients had trough levels above 1170 ng/ml (Q4). There were 31 patients (11.2%) with levels above 2000 ng/ml, all of whom were included in the Q4. Thirty seven patients in our cohort had more than one analysis of trough imatinib plasma levels done during the one year follow-up for a total of 82 analyses. Sub-group analysis of trough imatinib plasma levels was conducted in the 13 patients in the Q1 and the six patients in the Q4 who had 2 analyses done. Mean trough imatinib plasma levels went from 401ng/ml to 665 ng/ml in the Q1 patients and from 2845 ng/ml to 1065 ng/ml in the Q4 patients. These results confirm the feasibility of imatinib plasma levels testing in the community and the important variability of trough imatinib plasma concentrations in individual patients, as described by other groups. A significant portion of patients in our cohort had trough levels below 647 ng/ml, which has been associated with less favourable cytogenetic and molecular responses in studies. These results also suggest that physicians act on the information procured by the determination of imatinib plasma levels as second level determination was improved for patients initially in Q1 or Q4. Further follow up analyses are needed to document if optimisation of dosing leads to better response or improvement in tolerability of the drug.

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Macrophages and iron trafficking at the birth and death of red cells

Blood ◽

10.1182/blood-2014-12-567776 ◽

2015 ◽

Vol 125 (19) ◽

pp. 2893-2897 ◽

Cited By ~ 72

Author(s):

Tamara Korolnek ◽

Iqbal Hamza

Keyword(s):

Growth Factors ◽

Blood Cells ◽

Iron Homeostasis ◽

Critical Role ◽

The Body ◽

Red Cells ◽

Physical Contact ◽

Red Cell ◽

Iron Trafficking ◽

Intimate Association

Abstract Macrophages play a critical role in iron homeostasis via their intimate association with developing and dying red cells. Central nurse macrophages promote erythropoiesis in the erythroblastic island niche. These macrophages make physical contact with erythroblasts, enabling signaling and the transfer of growth factors and possibly nutrients to the cells in their care. Human mature red cells have a lifespan of 120 days before they become senescent and again come into contact with macrophages. Phagocytosis of red blood cells is the main source of iron flux in the body, because heme must be recycled from approximately 270 billion hemoglobin molecules in each red cell, and roughly 2 million senescent red cells are recycled each second. Here we will review pathways for iron trafficking found at the macrophage-erythroid axis, with a focus on possible roles for the transport of heme in toto.

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Vascular Growth Factors and Lymphangiogenesis

Physiological Reviews ◽

10.1152/physrev.00005.2002 ◽

2002 ◽

Vol 82 (3) ◽

pp. 673-700 ◽

Cited By ~ 260

Author(s):

Lotta Jussila ◽

Kari Alitalo

Keyword(s):

Endothelial Cells ◽

Growth Factors ◽

Lymphatic System ◽

Lymphatic Vessels ◽

The Body ◽

Vascular Tumors ◽

Vascular Endothelial ◽

Lymphatic Endothelial Cells ◽

Independent Manner ◽

Tumor Spread

Blood and lymphatic vessels develop in a parallel, but independent manner, and together form the circulatory system allowing the passage of fluid and delivering molecules within the body. Although the lymphatic vessels were discovered already 300 years ago, at the same time as the blood circulation was described, the lymphatic system has remained relatively neglected until recently. This is in part due to the difficulties in recognizing these vessels in tissues because of a lack of specific markers. Over the past few years, several molecules expressed specifically in the lymphatic endothelial cells have been characterized, and knowledge about the lymphatic system has started to accumulate again. The vascular endothelial growth factor (VEGF) family of growth factors and receptors is involved in the development and growth of the vascular endothelial system. Two of its family members, VEGF-C and VEGF-D, regulate the lymphatic endothelial cells via their receptor VEGFR-3. With the aid of these molecules, lymphatic endothelial cells can be isolated and cultured, allowing detailed studies of the molecular properties of these cells. Also the role of the lymphatic endothelium in immune responses and certain pathological conditions can be studied in more detail, as the blood and lymphatic vessels seem to be involved in many diseases in a coordinated manner. Discoveries made so far will be helpful in the diagnosis of certain vascular tumors, in the design of specific treatments for lymphedema, and in the prevention of metastatic tumor spread via the lymphatic system.

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Comparison of Release Profiles of Various Growth Factors from Biodegradable Carriers

MRS Proceedings ◽

10.1557/proc-530-13 ◽

1998 ◽

Vol 530 ◽

Cited By ~ 1

Author(s):

Y. Tabata ◽

M. Yamamoto ◽

Y. Ikada

Keyword(s):

Growth Factors ◽

Growth Factor ◽

Transforming Growth Factor ◽

Transforming Growth Factor Β1 ◽

The Body ◽

Bone Morphogenetic Protein 2 ◽

Biodegradable Hydrogel ◽

Tgf Β1

AbstractA biodegradable hydrogel was prepared by glutaraldehyde crosslinking of acidic gelatin with an isoelectric point (IEP) of 5.0 as a carrier to release basic growth factors on the basis of polyion complexation. Basic fibroblast growth factor (bFGF), transforming growth factor β1 (TGF-β1), and bone morphogenetic protein-2 (BMP-2) were sorbed from their aqueous solution into the dried gelatin hydrogels to prepare respective growth factor-incorporating hydrogels. Under an in vitro non-degradation condition, approximately 20 % of incorporated bFGF and TGF-β1 was released from the hydrogels within initial 40 min, followed by no further release, whereas a large initial release of BMP-2 was observed. After subcutaneous implantation of the gelatin hydrogels incorporating 125I-labeled growth factor in the mouse back, the remaining radioactivity was measured to estimate the in vivo release profile of growth factors. Incorporation into gelatin hydrogels enabled bFGF and TGF-β1 to retain in the body for about 15 days and the retention period well correlated with that of the gelatin hydrogel. Taken together, it is likely that the growth factors ionically complexed with acidic gelatin were released in vivo as a result of hydrogel biodegradation. On the contrary, basic BMP-2 did not ionically interact with acidic gelatin, resulting in no sustained released by the present biodegradable carrier system.

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