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Scanning and Transmission Electron Microscopy Reveals Graphene Oxide in CoV-19 Vaccines
... Por ejemplo, en los estudios de microscopía óptica sobre alícuotas del contenido de los viales de las mismas marcas analizadas en este estudio, se encontraron diversos tipos de micropartículas con tamaños en el rango de 1-500 μm, cuyas identidades y funciones se desconocen en su mayor parte. Entre las micro partículas halladas, se encontró grafeno o derivados del mismo como el óxido de grafeno o hidróxido de grafeno, que fueron identificados en partículas de ciertas características analizadas por la técnica de Micro-Raman y Microscopía Electrónica de Transmisión (TEM) (Campra, 2021;Young 2022). Este tipo de micro partículas ortogonales, se forman luego de transferir una parte del contenido de los viales en agua destilada o solución fisiológica, se originan a partir de otras partículas aún más pequeñas (de dimensiones nanométricas y compuestas en su mayor parte por los elementos que presentan distribución diferencial de muestreo) presentes en las fases del contenido de los viales pero que no pueden ser visualizadas con microscopía óptica. ...
... Estas micro partículas ortogonales se diferencian sustancialmente de cristales de sales orgánicas e inorgánicas por los siguientes motivos: el patrón temporal de su proceso de formación y las condiciones en las que este ocurre; sus morfologías no presentan estructuras geométricas de tipo fractales, que son características de los cristales de sales orgánicas e inorgánicas; y por sus elementos constituyentes que además de ser diferentes a aquellos de cristales de sales orgánicas o inorgánicas, se presentan en cantidades y distribuciones heterogéneas. Muchos de los elementos en estas micro estructuras particulares han sido identificados con un alto grado de precisión, mediante el uso de SEM-EDX, técnica que se basa en microscopía electrónica de barrido, para focalizar específicamente la microestructura que se quiere analizar, acoplada a dispersión de rayos X, para identificar los elementos que componen dicha microestructura (Martínez et al., 2021, Young, 2022, Nagase, 2022, Sangorrín y Diblasi, 2022b, Hagima, 2023. ...
Resumen: Como consecuencia de la elevada toxicidad de los productos inyectables, en fase experimental, denominados "vacunas contra COVID-19", cuyo uso ha sido impulsado vehementemente mediante una masiva campaña mundial de vacunación iniciada a finales del año 2020, la población global ha desarrollado y sufrido innumerables y variadas afecciones a su salud en grados leves, moderados y graves. El número de muertes y efectos adversos asociados a estos inyectables supera con creces a aquellos producidos por la suma de todas las vacunas anteriores. Al respecto, fue notable el aumento atípico de muertes súbitas y también de muertes provocadas por otras dolencias. Este incremento de trastornos en la salud de la población comenzó a manifestarse en concomitancia con el número de personas inoculadas y de dosis administradas por persona, afectando a la población inoculada en particular. Por lo cual, desde el año 2021 queda en evidencia que el incremento en la cantidad y variedad de patologías está claramente asociado a la aplicación de estos productos experimentales. En base a los 24 elementos químicos no declarados detectados hasta finales del año 2023, mediante el empleo de SEM-EDX y otras metodologías, por diferentes grupos de investigadores independientes, para obtener información más precisa sobre el contenido de los viales de las diferentes marcas de "vacunas contra COVID-19", y considerando el alcance limitado de cada metodología empleada para tal fin, el objetivo de este estudio fue corroborar estos hallazgos, identificar posibles elementos adicionales a los ya descubiertos y cuantificar la cantidad de todos los elementos encontrados. Para tal fin, se analizaron los contenidos de viales de diferentes lotes de las marcas AstraZeneca/Oxford, CanSino Biologics, Pfizer/BioNTech, Sinopharm, Moderna y Sputnik V. Se identificaron con gran precisión y se cuantificaron por ICP-MS 55 elementos químicos no declarados.
... But the aim of the COVID-19 mRNA vaccines is to reprogram the body's cells to produce a foreign Spike protein, or components of it, with unforeseen impact on the cells and organs of our body. Evidence is also rapidly accumulating to show that the experimental formulations, already injected into billions of human beings, appears to contain additional foreign particles that have been plausibly associated with problems in vital systems dependent on circulation of the blood Young, 2022;Benzi-Cipelli et al., 2022). The lack of transparency regarding the chemical, physical, nucleotidic and other contents of the injectable preparations is, to say the least, inexcusable. ...
... However, the Spike protein has been detected in the plasma of vaccine recipients (Ogata et al., 2022) which, considering that the Spike itself is supposedly a structurally stable protein (Lyons-Weiler, 2020; Vojdani & Kharrazian, 2020;Vojdani et al., 2021) with virus-independent cytotoxic effects (Pardi et al., 2015;Polack et al., 2020, Scioli Montoto et al., 2020 calls for the urgent need of pharmacovigilance studies. Moreover, foreign materials present in the vaccines have also been detected in the plasma of vaccine recipients as late as four months after the injection, reinforcing the need of reassessing the pharmacokinetics, the actual contents, and the clinically known impact of the products on recipients Benzi-Cipelli et al., 2022;Young, 2022). ...
In the context of mass vaccination campaigns, the most widely used vaccines in Western countries are based on messenger RNA (mRNA). Some countries have imposed mandatory vaccination and many others have required a vaccination passport to access public transportation and many activities, producing systemic discrimination, social exclusion, segregation, and stigmatization against non-vaccinated individuals. This paper aims to present several scientific uncertainties on which, conscientious objectors to mRNA injections as a preventive treatment for COVID-19, could rely. Scientific data are presented on mRNA vaccines, which consist in mRNAs wrapped in lipid nanoparticles. Never used as a prophylactic drug, artificial mRNAs delivered to our cells forces them to express, against their nature, a biologically active viral protein. Unlike a drug produced in a pharmaceutical factory and formulated at a known dose and a well-defined protein product profile, the mRNA vaccine acts as a pro-drug encoding for the viral Spike protein of the virus to be produced by our own cells; both the dose and the quality of the proteins produced are unknown. We also ignore the distribution of the lipid nanoparticles carrying this mRNA in our body. We consider that the “conscientious objection” raised by the above considerations is a reason enough to refuse mRNA vaccines or similar technologies as a preventive treatment against COVID-19.
... Apart from COVID-19 identification, SEM has been used with other microscopy in studying the vaccines. SEM coupled with an x-ray microprobe of an Energy Dispersive Spectroscopy (EDS) helped in the evaluation of particle size, composition distribution and chemical nature of vaccines developed by companies such as Pfizer, Moderna, Astrazeneca and Janssen [35]. ...
The humanity has witnessed a new virus called SARS-CoV-2 (also called as COVID-19) that emerged in 2019 causing the pandemic
... "Although information on the in vitro and in vivo nano toxicity of graphene nano materials has been increasingly published in the last several years, a complete picture on the bio-compatibility of graphene nano-materials has not been established. The successful applications of graphene nano-materials in nano bio-technology and medicine as well as their effective translation into real clinical utility hinge significantly on a thorough understanding of their nano toxicological profile [10]. Of all aspects of biocompatibility, the hemocompatibility of graphene nano materials with the different blood constituents in circulatory system is one of the most important elements that need to be well elucidated. ...
This work starts after seeing a recent open letter for transparency related production a quality control technique of m RNA vaccine first signed by Tarro G, Luisetto M and Monsellato ML and an Editorial recognized by IMA Marijnskaya academy: Graphene and Derivates: Physico-Chemical and Toxicology properties in the m-RNA Vaccine Manufacturing Strategy Needed specific proof of absence for the regulatory aspects (accepted for publication). Other relevant evidences come from the work of Giovannini et al. related Dark Field microscopy assay of the blood of 1086 symptomatic subjects after vaccination with two types of mRNA vaccine of great interest on this field the work of Campra P and Young RO, Young Me Lee or Ki-Yeob J. The Aim of this work is to investigate the self-auto assembly properties of graphene and derivates in order to Find relationship in some biotechnological application like mRNA vaccine. After a review part an experimental hypotesys project will be submitted to the researcher to produce a global conclusion. The recent evidences published in last period induced the idea to more deeply study these properties for The clinico-toxicological aspects involved.
ARTICLE
Graphene oxide (GO) suspensions in solvents are the most important feedstocks for preparing GO based composites, and the dispersion state of GO on the microscale in solvent is a dominating factor in determining the physical properties of GO based composites. However, the morphology of GO sheets in solvents has hardly been reported due to the limitation of the characterization methods. Here, we report that the sheet thickness and lateral size of GO in solution can be identified using optical microscopy (OM) within a couple of minutes. The dispersion states of GO, including stretched flakes, scrolls, crumbles, and agglomerates, can also be distinguished. Moreover, the dispersion states, which change with the pH value and ionic strength of the solvent, are closely related to the dispersion stability of the GO suspension and the morphology of the GO/PVA composite. We believe that the fast observation and identification of GO sheets and their structural features in solvents, enabled by OM, opens up a new avenue to studying GO based composite materials in liquids.
Natural graphite is an inexpensive and abundant source to obtain graphene nanosheets. The most efficient method for large-scale production is the chemical method, which is based on the oxidation of natural graphite. This paper reports the synthesis and characterization of graphene obtained by the Hummers method with some modifications. The results indicate a high degree of graphite oxidation, proving that the process was efficient. Analyses of field emission scanning electron microscopy (FEG), transmission electron microscopy (TEM), Raman spectroscopy, thermogravimetric analysis (TGA) and X-ray diffraction showed that the graphene produced presented characteristics similar to the commercial graphene.
Due to their unique physicochemical properties, graphene-family nanomaterials (GFNs) are widely used in many fields, especially in biomedical applications. Currently, many studies have investigated the biocompatibility and toxicity of GFNs in vivo and in intro. Generally, GFNs may exert different degrees of toxicity in animals or cell models by following with different administration routes and penetrating through physiological barriers, subsequently being distributed in tissues or located in cells, eventually being excreted out of the bodies. This review collects studies on the toxic effects of GFNs in several organs and cell models. We also point out that various factors determine the toxicity of GFNs including the lateral size, surface structure, functionalization, charge, impurities, aggregations, and corona effect ect. In addition, several typical mechanisms underlying GFN toxicity have been revealed, for instance, physical destruction, oxidative stress, DNA damage, inflammatory response, apoptosis, autophagy, and necrosis. In these mechanisms, (toll-like receptors-) TLR-, transforming growth factor β- (TGF-β-) and tumor necrosis factor-alpha (TNF-α) dependent-pathways are involved in the signalling pathway network, and oxidative stress plays a crucial role in these pathways. In this review, we summarize the available information on regulating factors and the mechanisms of GFNs toxicity, and propose some challenges and suggestions for further investigations of GFNs, with the aim of completing the toxicology mechanisms, and providing suggestions to improve the biological safety of GFNs and facilitate their wide application.
The knowledge on potential harmful effects of metallic nanomaterials lags behind their increased use in consumer products and therefore, the safety data on various nanomaterials applicable for risk assessment are urgently needed. In this study, 11 metal oxide nanoparticles (MeOx NPs) prepared using flame pyrolysis method were analyzed for their toxicity against human alveolar epithelial cells A549, human epithelial colorectal cells Caco2 and murine fibroblast cell line Balb/c 3T3. The cell lines were exposed for 24 h to suspensions of 3-100 μg/mL MeOx NPs and cellular viability was evaluated using. Neutral Red Uptake (NRU) assay. In parallel to NPs, toxicity of soluble salts of respective metals was analyzed, to reveal the possible cellular effects of metal ions shedding from the NPs. The potency of MeOx to produce reactive oxygen species was evaluated in the cell-free assay. The used three cell lines showed comparable toxicity responses to NPs and their metal ion counterparts in the current test setting. Six MeOx NPs (Al2O3, Fe3O4, MgO, SiO2, TiO2, WO3) did not show toxic effects below 100 µg/mL. For five MeOx NPs, the averaged 24 h IC50 values for the three mammalian cell lines were 16.4 µg/mL for CuO, 22.4 µg/mL for ZnO, 57.3 µg/mL for Sb2O3, 132.3 µg/mL for Mn3O4 and 129 µg/mL for Co3O4. Comparison of the dissolution level of MeOx and the toxicity of soluble salts allowed to conclude that the toxicity of CuO, ZnO and Sb2O3 NPs was driven by release of metal ions. The toxic effects of Mn3O4 and Co3O4 could be attributed to the ROS-inducing ability of these NPs. All the NPs were internalized by the cells according to light microscopy studies but also proven by TEM, and internalization of Co3O4 NPs seemed to be most prominent in this aspect. In conclusion, this work provides valuable toxicological data for a library of 11 MeOx NPs. Combining the knowledge on toxic or non-toxic nature of nanomaterials may be used for safe-by-design approach.
Dispersions of few-layer (1-3 layers), multi-layer (4-10 layers) and thick-layer (>10 layers) graphene oxide (GO) were prepared by a modified Hummers method with different mass ratios of KMnO4 to graphite. Ultraviolet-visible (UV-vis) spectroscopic data show that few-layer GO dispersions can be distinguished from multi- and thick-layer dispersions by a more intense peak at 230 nm. Atomic force microscopy (AFM) images of few-layer GO contain a single peak, those of multi-layer GO exhibit a shoulder and those of thick-layer GO do not contain a peak or shoulder. These findings allow qualitative analysis of GO dispersions. X-ray photoelectron spectra (XPS) show that the change of UV-vis absorption intensity of GO is caused by a conjugative effect related to chromophore aggregation that influences the π-π* plasmon peak.
Graphene-based sheets such as graphene, graphene oxide and reduced graphene oxide have stimulated great interest due to their promising electronic, mechanical and thermal properties. Microscopy imaging is indispensable for characterizing these single atomic layers, and oftentimes is the first measure of sample quality. This review provides an overview of current imaging techniques for graphene-based sheets and highlights a recently developed fluorescence quenching microscopy technique that allows high-throughput, high-contrast imaging of graphene-based sheets on arbitrary substrate and even in solution.
Nanoparticles (NPs) are tiny materials used in a wide range of industrial and medical applications. Titanium dioxide (TiO(2)) is a type of nanoparticle that is widely used in paints, pigments, and cosmetics; however, little is known about the impact of TiO(2) on human health and the environment. Therefore, considerable research has focused on characterizing the potential toxicity of nanoparticles such as TiO(2) and on understanding the mechanism of TiO(2) NP-induced nanotoxicity through the evaluation of biomarkers. Uncoated TiO(2) NPs tend to aggregate in aqueous media, and these aggregates decrease cell viability and induce expression of stress-related genes, such as those encoding interleukin-6 (IL-6) and heat shock protein 70B' (HSP70B'), indicating that TiO(2) NPs induce inflammatory and heat shock responses. In order to reduce their toxicity, we conjugated TiO(2) NPs with polyethylene glycol (PEG) to eliminate aggregation. Our findings indicate that modifying TiO(2) NPs with PEG reduces their cytotoxicity and reduces the induction of stress-related genes. Our results also suggest that TiO(2) NP-induced effects on cytotoxicity and gene expression vary depending upon the cell type and surface modification.
The Nuremberg Code drafted at the end of the Doctor's trial in Nuremberg 1947 has been hailed as a landmark document in medical and research ethics. Close examination of this code reveals that it was based on the Guidelines for Human Experimentation of 1931. The resemblance between these documents is uncanny. It is unfortunate that the authors of the Nuremberg Code passed it off as their original work. There is evidence that the defendants at the trial did request that their actions be judged on the basis of the 1931 Guidelines, in force in Germany. The prosecutors, however, ignored the request and tried the defendants for crimes against humanity, and the judges included the Nuremberg Code as a part of the judgment. Six of ten principles in Nuremberg Code are derived from the 1931 Guidelines, and two of four newly inserted principles are open to misinterpretation. There is little doubt that the Code was prepared after studying the Guidelines, but no reference was made to the Guidelines, for reasons that are not known. Using the Guidelines as a base document without giving due credit is plagiarism; as per our understanding of ethics today, this would be considered unethical. The Nuremberg Code has fallen by the wayside; since unlike the Declaration of Helsinki, it is not regularly reviewed and updated. The regular updating of some ethics codes is evidence of the evolving nature of human ethics.
After uptake into target cells, many nonenveloped viruses undergo conformational changes in the low-pH environment of the endocytic compartment. This results in exposure of amphipathic viral peptides and/or hydrophobic protein domains that are inserted into and either disrupt or perforate the vesicular membranes. The viral nucleic acids thereby gain access to the cytosol and initiate replication. We here demonstrate the in vitro transfer of the single-stranded positive-sense RNA genome of human rhinovirus 2 into liposomes decorated with recombinant very-low-density lipoprotein receptor fragments. Membrane-attached virions were exposed to pH 5.4, mimicking the in vivo pH environment of late endosomes. This triggered the release of the RNA whose arrival in the liposomal lumen was detected via in situ cDNA synthesis by encapsulated reverse transcriptase. Subsequently, cDNA was PCR amplified. At a low ratio between virions and lipids, RNA transfer was positively correlated with virus concentration. However, membranes became leaky at higher virus concentrations, which resulted in decreased cDNA synthesis. In accordance with earlier in vivo data, the RNA passes through the lipid membrane without causing gross damage to vesicles at physiologically relevant virus concentrations.
Quantum Dots (QDs), are considered as promising tools for biomedical applications. They have potential applications in agricultural industries, novel pesticide formulations, use in bio-labels and devices to aid genetic manipulation and post-harvest management. Since interactions with higher plants are of important environmental and ecological concern we investigated the cytotoxicity and genotoxicity of CdSe QDs in a model plant (Allium cepa) and established relationships between QDs genotoxic activity and oxidative stress.
Allium cepa bulbs with intact roots were exposed to three concentrations of CdSe QDs (12.5, 25 and 50 nM). Cell viability and mitotic frequencies was measured for cytotoxicity, and to assess the genotoxicity DNA lesions, chromosome aberrations and micronuclei were evaluated. We report that QDs exerted significant genotoxic effects, associated with oxidative stress. This could be correlated with the retention of Cd in Allium roots as a dose-dependent increase with the highest uptake at 50 nM of CdSe QD. Oxidative stress induced by CdSe QD treatment activated both, antioxidant (SOD, CAT) scavengers and antioxidant (GPOD, GSH) enzymes. Concentrations as low as 25 nM CdSe QDs were cytotoxic and 50 nM CdSe QDs was found to be genotoxic to the plant. These findings enable to determine the concentrations to be used when practical applications using nanodevices of this type on plants are being considered.
The US Food and Drug Administration (FDA) Expanded Access (EA) Program, which allows for compassionate uses of unapproved therapeutics and diagnostics outside of clinical trials, has gained significant traction during the Coronavirus 2019 (COVID-19) pandemic. While development of vaccines has been the major focus, uncertainties around new vaccine safety and effectiveness have spawned interest in other pharmacological options. Experimental drugs can also be approved under the FDA Emergency Use Authorization (EUA) program, designed to combat infectious disease and other threats. Here, we review the US experience in 2020 with pharmacological EA and EUA approvals during the pandemic. We also provide a description of, and clinical rationale for, the EA- or EUA-approved drugs during the pandemic and concluding reflections on the EA program and its potential future uses.
The nanotechnology enabled the development of nanomaterials (NMs) with a variety of industrial, biomedical, and consumer applications. However, the mechanism of action (MoA) and toxicity of NMs remain unclear, especially in the male reproductive system. Thus, this study aimed to perform a bibliometric and systematic review of the literature on the toxic effects of different types of NMs on the male reproductive system and function in mammalian models. A series of 236 articles related to the in vitro and in vivo reproductive toxicity of NMs in mammalian models were analyzed. The data concerning the bioaccumulation, experimental conditions (types of NMs, species, cell lines, exposure period, and routes of exposure), and the MoA and toxicity of NMs were summarized and discussed. Results showed that this field of research began in 2005 and has experienced an exponential increase since 2012. Revised data confirmed that the NMs have the ability to cross the blood-testis barrier and bioaccumulate in several organs of the male reproductive system, such as testis, prostate, epididymis, and seminal vesicle. A similar MoA and toxicity were observed after in vitro and in vivo exposure to NMs. The NM reproductive toxicity was mainly related to ROS production, oxidative stress, DNA damage and apoptosis. In conclusion, the NM exposure induces bioaccumulation and toxic effects on male reproductive system of mammal models, confirming its potential risk to human and environmental health. The knowledge concerning the NM reproductive toxicity contributes to safety and sustainable use of nanotechnology.
Vaccine is one of the most effective strategies for preventing and controlling infectious diseases and some noninfectious diseases, especially cancers. Adjuvants and carriers have been appropriately added to the vaccine formulation to improve the immunogenicity of the antigen and induce long-lasting immunity. However, there is an urgent need to develop new all-purpose adjuvants because some adjuvants approved for human use have limited functionality. Graphene oxide (GO), widely employed for the delivery of biomolecules, excels in loading and delivering antigen and shows the potentiality of activating the immune system. However, GO aggregates in biological liquid and induces cell death, and it also exhibits poor biosolubility and biocompatibility. To address these limitations, various surface modification protocols have been employed to integrate aqueous compatible substances with GO to effectively improve its biocompatibility. More importantly, these modifications render functionalized-GO with superior properties as both carriers and adjuvants. Herein, the recent progress of physicochemical properties and surface modification strategies of GO for its application as both carriers and adjuvants is reviewed.
Statement of Significance
Due to its unique physicochemical properties, graphene oxide is widely employed in medicine for purposes of photothermal treatment of cancer, drug delivery, antibacterial therapy, and medical imaging. Our work describes the surface modification of graphene oxide and for the first time summarizes that functionalized graphene oxide serves as a vaccine carrier and shows significant adjuvant activity in activating cellular and humoral immunity. In the future, it is expected to be introduced into vaccine research to improve the efficacy of vaccines.
A new approach to drug design based on nanoparticles and related nanostructures for effective drug delivery, is of great importance in future medical treatment, especially for cancer therapy. Nanomaterials hold tremendous potential for increasing the efficiency of drug delivery, with a high degree of biocompatibility. Additionally, for biomedical applications, they must be biodegradable, have prolonged circulation half-life, not tend to aggregate or cause an inflammatory response in the body and to be cost-effective. The efficacy of such structures is highly dependent on their chemical properties as well as on shape, charge, size, surface modifications and loading method. Here we focused on the potential of using different kinds of nanoparticles and similar nanostructures loaded with various drugs in order to achieve specific targeting and controlled drug release. Thereby, computational modeling on NPs-based drug delivery could help in providing a better understanding of all parts of the delivery system. This review emphasizes recent advances in the usage of various types of nanoparticles and similar nanostructures for drug delivery, aiming to provide a critical review of less toxic and more effective treatment.
Recently, medical research has been shifting its focus to nanomedicine and nanotherapeutics in the pursuit of drug development research. Quantum dots (QDs) are a critical class of nanomaterials due to their unique properties, which include optical, electronic, and engineered biocompatibility in physiological environments. These properties have made QDs an attractive biomedical resource such that they have found application as both in vitro labeling and in vivo theranostic (therapy-diagnostic) agents. Considerable research has been conducted exploring the suitability of QDs in theranostic applications, but the cytotoxicity of QDs remains an obstacle. Several types of QDs have been investigated over the past decades, which may be suitable for use in biomedical applications if the barrier of cytotoxicity can be resolved. This review attempts to report and analyze the cytotoxicity of the major QDs along with relevant related aspects.
We demonstrated a way to achieve tunable fluorescence from reduced graphene oxide (rGO) by selecting appropriate solvents. This work begins a new area of research that seeks to improve the fluorescence properties of rGO, offering novel strategies for realizing printed flexible opto-electronics with tunable device performance. We investigated the effect of solvents and report that solvents play a vital role in the optical properties of rGO. The dispersion of rGO was analyzed by UV–Visible spectroscopy, FTIR spectroscopy, and fluorescence spectroscopy, demonstrating that rGO dissolved in polar solvents exhibits enhanced fluorescence covering the entire visible range as opposed to non-polar solvents. The synthesis strategy may possibly facilitate the manipulation and processing of rGO toward vital printed flexible optoelectronics applications, which could be tuned by selecting a suitable solvent.
A nano-graphene bilayer is described by a transverse Ising model with single-ion anisotropy. The upper layer consists of spin 3/2 with an antiferromagnetic exchange coupling, whereas the bottom layer composes of spin 5/2 with a ferromagnetic exchange coupling. Based on the effective-field theory with correlations the general formula for the magnetization is given. Magnetic properties of the system have been studied, such as the magnetization, the magnetic susceptibilities and the blocking temperature. The exchange coupling, the single-ion anisotropy, the transverse field and the external magnetic field have important effects on the blocking temperature.
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