Haha - we have to hire an attorney to go to trial and help structure and argue the main case too (why we are fundraising)- Its so aggravating to have to teach them and prod them and literally do ALL the work because in general they do the very MINIMUM... ughhhh. Whats your case in NY? Hope you win!!!!
Slow is always more thorough! We are praying for a fast win Nov 9 though to at least stop the shots locally in CR but hopefully globally. The laws here do allow the CR judge to have jurisdiction over the "superiors", in this case: WHO, FDA, CSC, EMA, etc. PRAY!!!!
What about the full list of ingredients for these poisonous injections?
It's easy to explain why they're toxic if they were exposed.
And preventing people from knowing what causes harm and, consequently, what can prevent it, what is an antidote to that toxicity, could save lives and prevent so many injuries...
Isn't that a crime???
Hiding the truth about the toxicity of ALL the ingredients?
Please see this post for information as it is the same technology:
1. Contentious Administrative (the Nov 9 hearing) to prove its outside the limits of the law and VOID
2. Its criminal for the reasons you said and much more - Investigations are in process with parts concluded that are making their way to the correct court.
It should be investigated whether TOXIC NANOTECHNOLOGY used to "prevent" "Covid" (in masks and tests and respiratory filters and many drugs, including Remdesivir) can cause the so-called Covid and can cause injury and death:
“A recent study of particular objects known as "nanotubes," revered for their extraordinary strength and electrical conductivity, demonstrated that such objects tend to clump within the lungs, causing suffocation.”
“Take the experience of researchers at DuPont, who are testing microscopic tubes of carbon, known as nanotubes, valued for their extraordinary strength and electrical conductivity.
When the researchers injected nanotubes into the lungs of rats in the summer of 2002, the animals unexpectedly began gasping for breath. Fifteen percent of them quickly died. ''It was the highest death rate we had ever seen,''
said David B. Warheit, the research leader, who began his career studying asbestos and has been testing the pulmonary effects of various chemicals for DuPont since 1984.
Early research has raised troubling issues. DuPont and others, for example, found evidence that the cells that break down foreign particles in rodent lungs have more trouble detecting and handling nanoparticles than larger particles that have long been studied by air pollution experts.
Lungs are not the only concern.
Research shows that nanoparticles deposited in the nose can make their way directly into the brain.
They can also change shape as they move from liquid solutions to the air, making it harder to draw general conclusions about their potential impact on living things. “
“Despite these potential benefits, the toxicity of CNTs (Carbon Nanotubes) is a major concern that needs to be more clearly understood and addressed. Pristine, water-insoluble CNTs have been found to be highly toxic in vitro to many different types of cells, including human keratinocytes, rat brain neuronal cells, human embryonic kidney cells, and human lung cancer cells. In addition, unmodified CNTs administered intratracheally to mice have been reported to induce the formation of lung granulomas. CNTs have also been shown to promote the aggregation of human platelets in vitro, and analogous carbon particulate matter found in the environment enhanced experimentally induced vascular thrombosis in rats.”
“[in 2015] Günter Oberdörster and co-authors published what is possibly the most comprehensive review of carbon nanotube toxicology studies to date. Focusing on inhalation of nanotubes, they document evidence of transient pulmonary inflammation, and rapid and persistent development of granulomatous lesions and interstitial fibrosis on exposure to single- and multiwalled carbon nanotubes in rodents. They cite data indicating that inhaled long and thin multiwalled carbon nanotubes can move to the lining surrounding the lungs and penetrate it, where they can potentially cause mesothelioma. Furthermore, the authors indicate that carbon nanotubes can act as a cancer promoter — with inhalation increasing the probability of developing lung cancer from exposure to other carcinogens.”
Li et al. (2007a) compared the pulmonary response of mice exposed to purified MWCNT by IT instillation versus inhalation.
Intratracheal instillation produced inflammation and severe destruction of alveolar structures, while inhalation predominately resulted in moderate pathology consisting of alveolar wall thickening and cell proliferation but general alveolar structure was retained. This study demonstrated significant differences in the type and degree of pulmonary responses to MWCNT in mice between bolus-type IT instillation and inhalation, with higher doses deposited in lung by inhalation resulting in only moderate effects compared to severe lesions induced by instillation of lower doses. (!!!!!) - PCR "TEST"!!!! This is why it is done close to brain not close to lungs!!!!
But there is a reason why SOME people die and get hurt by the so-called. "Covid." Why would someone inject/instill TOXIC nanotechnology/graphene/ethylene oxide through a so-called "PCR test"?
Why would someone add graphene and titanium oxide to make people inhale in so-called "masks"?
And why, the hell, add this nanotechnology to saline solution, "Remdesivir" (Veklury), "monoclonal antibodies", "C-19" injections (so-called "vaccines"), and all those others that are in the pipeline??? (Such as N-Plate - radiation poisoning "vaccine", RiVax - RW Malone "invention" against "castor poisoning", MarVax and other toxic injections against "Marburg", Ebola and so on???
So, although yes, there was no pandemic, toxic nanotechnology is killing and injuring people, causing "flu-like symptoms" and causing the FEAR of SUFFOCATION. (low saturation because of free and oxy radical, caused by oxidative stress/disbalance/nanotechnology toxicity...
CAN YOU PLESE ADDRESS THE TOXICITY OF NANOTECHNOLOGY IN YOUR STATEMENTS??? People listen to you. Can you take a look at this? As a drug maker you understand toxicology, right???
THEY KNOW WHAT DAMAGE these injections can cause!!!
They know what to use to mitigate the damage, although healthy people (and indeed no one) should get these injections – but they know how to prevent damage IF THESE NANOTECHNOLOGIES ARE INJECTED!!!
After all, ordinary people DO NOT EVEN KNOW WHAT they were injected with, so HOW SHOULD THEY KNOW what adverse effects these injections can cause and why people may die after these injections - and THEY HAVE NO IDEA how to save themselves!!!
4.2.7. Immune In the immune system, nanoparticle immunotoxicity refers to the adverse effects such as complement activation-mediated pseudoallergy, hypersensitivity, immunosuppression, and inflammasome effects [199]. In general, most nanoparticles tend to accumulate in organs of the mononuclear phagocytic system such as the liver and spleen. The immune cells that in turn produce toxic effects upon nanoparticle exposure include monocytes, platelets, leukocytes, dendritic cells, and macrophages [199].
Coating AuNPs with polyethylene glycol (PEG) avoided immunotoxic responses [199,200]. One study compared the immunotoxic effects of coating AuNPs with PEG and chicken ovalbumin (OVA) [200]. No significant cytotoxicity to RAW264.7 macrophages was observed at AuNP concentrations of 20 µg/mL. However, the uptake capacity for the OVA-AuNPs was greater compared to PEG-coated AuNPs. Additionally, PEG-coated AuNPs did not induce a significant increase in TNF-a, IL-6 and IL-1B for AuNPs larger than 35 nm. In general, small nanoparticles, despite differences in surface coating, appeared to present greater immunotoxic effects compared to larger ones [200]. AuNP toxicity in murine and human lymphocytes has showed overall viability was only significantly reduced at 200 µg/mL, but not any concentration below this [201]. Another key immune cell, dendritic cells have been targeted as points of entry for immunotherapeutic agents using AuNPs. Research has indicated that dendritic cells have very low cytotoxicity upon exposure to different sizes and concentrations of AuNPs [202]. However, small AuNPs with size of 10 nm displayed weak apoptotic effects in dendritic cells compared to larger nanoparticles. In terms of surface coating, positive charged-polymer-coated AuNPs did have a significant cytotoxic effect [202]. This indicates the necessity for correct identification of surface chemistry when designing biocompatible nanoparticles for cancer therapy and diagnosis usage, among other medical uses.
On the other hand, the immunotoxicity profile of IONPs is different. Within macrophages, RAW264.7 macrophages treated with IONPs demonstrated an increase in oxidative stress and an increase in cell proliferation within 24 h [203]. Further, another study indicated that murine and human macrophage cell lines exposed to PEI-coated IONPs induced the activation of toll-like receptor 4 signaling and ROS production via different pathways that in turn further increased the overall activation of macrophages leading to pro-inflammatory effects [204]. With B and T lymphocytes, the effect of IONPs remains unclear as initial studies indicated no effect on function and cell viability. While the lack of significant cytotoxicity still holds true with current studies, changes have been observed in T-cell function including delays in proliferation rate [205]. However, it has been demonstrated that compared to control at 13 weeks post-IONP injection, the distribution of B cells decreased while T cells increased. In addition, the number of dendritic cells increased, though surface markers for antigen presentation such as CD40 were decreased [206]. This suppression of antigen presentation in dendritic cells was a common feature indicated in other studies [150]. Thus, with dendritic cells, cytotoxicity was not a significant feature, but functional impairment was present.
Overall, the effects of nanoparticle toxicity on the immune system are complex and multi-faceted as it involves a variety of different cell types across organ systems. In general, the current research suggests that AuNPs present limited toxicity to the cells of the immune system including macrophages, lymphocytes, and dendritic cells. However, the results with IONPs s are more mixed, with some functional impairment effects demonstrated, though the cytotoxic potential to immune cells remains low. Additionally, the surface coating and electrostatic charge of the nanoparticles play a key role in immunotoxicity profile and potential evasion of the immune system responses in targeted drug-delivery systems, an area for additional research.
etc.
It is also known that antioxidants can prevent oxidative imbalances and oxidative damage, which can cause death, blood clots, organ damage and other adverse effects.
4.2.5. Pulmonary/Respiratory Nanoparticles are employed in different types of lung cancers as a method for targeted drug-delivery in therapy. Various research studies have examined the pulmonary toxicity associated with nanomedicine [187,188,189,190]. There seems to a degree of variability in the toxicity of AuNPs compared with other nanoparticle types, which appeared to be more consistent in their behavior [187,188]. A study by Avalos et al. compared the relative toxicity of silver- and gold-based nanoparticles on human pulmonary fibroblasts [187]. In general, the cytotoxic effect on the pulmonary fibroblasts was not size dependent for AuNPs, unlike some of the previous studies highlighted in sections on other organ systems. All sizes studied (30, 50, and 90 nm) demonstrated a reduction in cell mitochondrial activity and lactate dehydrogenase (LDH) leakage. Furthermore, in comparison to AgNPs, oxidative stress and production of ROS was greater with AuNPs in pulmonary fibroblast cells [188]. Another study examined three different human lung epithelial cell types (A549, BEAS-2b, and NHBE) for cytotoxic effects of AuNPs and AgNPs [188]. AuNPs were coated with either sodium citrate or chitosan, which created different surface charges on the particles. In general, A549 and BEAS-2B cells exhibited the least cytotoxic effects with an increase in LDH release only at the highest concentration of chitosan-coated AuNPs. However, NHBE cells were more affected in terms of cytotoxicity by AgNPs and AuNPs as measured by LDH release and membrane leakage [188].
Other researchers have investigated the cytotoxic effect of nanoparticles on key cells involved in the blood-air barrier in the pulmonary system [189,190]. In general, for different metal-organic frameworks, lung epithelial and alveolar macrophage cell lines were more adversely affected by lipid-coated nanoparticle systems [189]. Specifically, another study focused on an in vitro 3D lung model with three cell types of the epithelial tissue barrier: monolayer of alveolar cells, macrophages, and dendritic cells [190]. After initial exposure to AuNPs, there was no observable change in the cell morphology compared to control across the cell types. However, long-term effects are the current limitation of these research studies and remain unknown.
The general toxicity profile of IONPs in the pulmonary system is attributed to increased oxidative stress due to particle internalization, dissolution, release, and disruption of regular iron homeostasis [191]. In vivo studies have indicated that exposure to IONPs induces an elevated acute inflammatory response, which persists up to 28 days post-exposure. In addition, there was found to be an increase in the heat shock proteins and matrix metalloproteinases and evidence of progression to granulomas [191]. Additional research has focused on the toxic effects of metal nanoparticles following inhalation and intratracheal instillation as also are being used in the realm of targeted drug deliver in cancer [192,193]. After inhalational exposure of IONPs, there was a transient increase in acute total cell and neutrophil counts, pro-inflammatory chemokines, and oxidative stress in the initial time points. However, in the long-term there was no persistent inflammation at the end of the study demonstrating that IONPs had limited toxicity in the long-term [192].
Overall, while AuNPs appear to generally have adverse effects to the respiratory system, their significance in terms of long-term pulmonary cytotoxicity still varies depending on the type of AuNP and the particular lung cell type. Further studies are necessary to establish more generalized outlook on the effects of IONPs to various lung cells.
4.2.6. Reproductive Reproductive toxicity due to nanoparticles require further differentiation between their effects on male versus female reproductive systems. In general, nanoparticles have been demonstrated to cross the blood-testicle and blood-placenta barrier, pressing the importance of addressing reproductive toxicity [194]. Some studies have shown a decrease in sperm motility, albeit only at very high concentrations of AuNPs.
Female: Within the female reproductive system, nanoparticle accumulation tends to occur in the uteri and the ovaries [195]. In the case of AuNPs and IONPs, the greater accumulation within the uterus was observed for smaller nanoparticles. In addition, the negative impact on female sex hormones was largely seen in current studies with titanium nanoparticles (TiO2) as they increased expression of the Cyp17a1 gene which in turn increased estradiol, apoptotic-related genes, inflammatory and immune responses, among other effects [195]. Furthermore, some nanoparticles have shown to produce morphological changes in the follicles leading to a reduction in the mature oocytes present [196]. Ovarian toxicity was observed with long-term TiO2 nanoparticle, which caused a shapeless follicular antrum and irregular arrangement of cells, though, the results were inconclusive and not very general [196].
Male: In the male reproductive system, nanoparticle exposure tends to not only have an impact on the reproductive organs themselves, but also some potential effects on spermatogenesis and motility [195]. One particular study with AuNS found limited toxicity to the testes with no necrosis or histological disorganization within the germinal cells, spermatozoids, intertubular spaces, Leydig cells, and Sertoli cells in male mice [197]. On the other hand, additional research has indicated that exposure to zinc oxide nanoparticles (ZnO) presented with a reduction in testicular tissue and loss of cells in seminiferous tubules at an intraperitoneal dose of 250, 500, and 700 mg/kg/day [185]. Testicular toxicity due to ZnO has been established by several studies and some work has characterized the particular changes that manifest in the blood-testis barrier as well [196]. Mechanistically, ZnO have been suggested to trigger ROS, potentiate DNA lesions in germ cells, and downregulate the expression of gap junction proteins in the cell membrane [196]. Other researches have employed ligand-free and oligonucleotide-conjugated AuNPs to study toxic effects on spermatozoa specifically [198]. The findings indicated that sperm morphology and viability was generally not affected at any concentration [198].
In summary, within the male and female reproductive system, there is a limitation in current research on the cytotoxic potential of nanoparticles. The current studies seem to indicate that there are certain established adverse effects as those described in the spermatozoa and hormonal changes. However, the morphological changes to the reproductive organs and their long-term implications are highly dependent on the nanoparticle type, coating, and the cell type affected as with other organ systems [196].
4.2.3. Nervous The nervous system is complex with many nuances that are not yet elucidated within the system itself, especially the brain. However, several research studies have examined aspects of nanotoxicity of AuNPs and IONPs, which will be the focus of this subsection.
Siddiqi et al. investigated changes in several biomarkers indicative of neurotoxicity upon injection of AuNPs in rat brains [174]. On one hand, there was a significance decrease in the enzyme glutathione peroxidase, which is an antioxidant in the brain that protects against oxidative damage. On the other hand, there were increases in markers of oxidative stress-derived DNA damage such as 8-hydroxydeoxyguanosine and heat shock protein 70 as well as apoptotic markers such as caspase-9. Furthermore, there was a significant increase in the neurotransmitters dopamine and serotonin, which demonstrated implications of AuNPs in potentiation of mood disturbances and chemical imbalances in the brain [174]. Other research has specifically studied the impact of AuNPs of different types on neural cells through the blood-retinal barrier [175]. In general, only small particles (less than 20 nm) were able to pass through the blood-retinal barrier and accumulate in the retinal layers. Furthermore, there was no toxicity demonstrated to neural cells such as retinal astrocytes, neurofilaments, and retinoblastoma cells in C57BL/6 mice [175].
The variability of toxicity effects on neural cells greatly depends upon the cell type and composition of the nanoparticles, which was demonstrated in other organ systems as well. For example, Joris et al. studied six different cell lines that included human and murine neural stem cells, human and mouse-derived progenitor cell lines, and human and murine neuroblastoma cell lines [176]. The study compared the relative toxicity effects of gold, iron oxide, and silver nanoparticles among these cell lines. Overall, it was found that AuNPs had the greatest degree of acute toxicity and IONPs having the lowest cytotoxic potential. In terms of the cell morphology, the C17.2 cell line was the only one with a reduction in cell area.
The cytotoxic potential of IONPs was further investigated on cultured neurons, astrocytes, and microglial cells. In rat cerebellar granular neurons, dimercaptosuccinate-coated IONPs accumulated within the cultured neurons by 1000-fold, but cellular integrity or viability was not adversely affected. This finding indicated the inherent potential of neurons to mediate oxidative stress effects through their antioxidant abilities [177]. While the finding with neurons was consistent with astrocytes, it did not hold true for microglial cells, which were rapidly damaged and displayed severe toxicity. At a mechanistic level, the rate of release of iron from internalized IONPs in microglial cells may be too rapidly transferred to lysosomes leading to toxic iron levels within the cells. Similarly, the cytotoxic potential of nanoparticles varies based on the particular mechanistic pathways associated with different nervous cell types to handle oxidative stress induced by the nanoparticles [176,177]. From a pathological perspective, accumulation of iron in the brain has been linked to neurodegenerative diseases such as aceruloplasminemia and neuroferritinopathy, as well as having the potential to play a role in Alzheimer’s and Parkinson’s diseases [178]. This research emphasizes the importance of finding the optimal amount of IONPs for therapeutic purposes in the nervous system.
4.2.4. Cardiovascular Cardiovascular nanomedicine has been employed in the diagnosis and treatment of cardiovascular diseases in addition to its role in cancer therapy [179]. One application of AuNPs in cancer therapy is the loading of these particles with doxorubicin (DOX, chemotherapy drug) for targeted drug delivery. The cardiotoxicity of these DOX-loaded and PEG-coated AuNPs have been studied in current pharmacological research [180]. Ultimately, DOX loaded onto AuNPs demonstrated no significant changes in cardiovascular function biomarkers such as serum lactate dehydrogenase (LDH) and creatinine kinase MB (CK-MB) levels compared to the free DOX. Similarly, another study further solidified these results in that DOX loaded on AuNPs did not create changes in CK-MB levels compared to baseline [181]. CK-MB is an enzyme in the myocardium that has served as the gold standard indicator of myocyte injury in many clinical and research settings [181].
There is limited research on the chronic cardiac toxicity of AuNPs, which was the aim of a study conducted by Yang et. al. on the effect of PEG-coated AuNPs at 2, 4, and 12-week time periods [182]. There was no significant decrease in the left ventricular ejection fraction across each time point for all sizes studied. Inflammatory mediators such as CD45+ and TNF-a indicated that chronic exposure to AuNPs did not spark inflammatory cell infiltration in the heart. Other studies that employed PEG-coated AuNPs have also concluded upon similar results that accumulation of AuNPs in the heart did not induce significant changes in cardiac hypertrophy, fibrosis or inflammation further demonstrating the strong biocompatibility of PEG-coated AuNPs in biomedical applications [183].
In the context of IONPs, cardiotoxic effects have been observed with IONPs in connection to myocardial damage due to iron accumulation. Unlike in the spleen, the macrophage clearance ability in the heart is limited, leading to longer-term accumulation not fully characterized in present research [184]. Particularly, some research has indicated that IV administration of IONPs resulted in a pro-coagulatory effect in vivo and in vitro while causing oxidative stress on the heart [185]. The effect of IONPs on different cardiac markers of oxidative stress in mice was also investigated. Data has indicated a significant increase of lipid peroxidation, reactive oxygen species, and superoxide dismutase in heart tissue compared with control groups [186].
Overall, there seems to be a certain degree of cardiotoxicity associated with IONPs, but the systemic effect of these microscopic changes is yet to fully be determined by current research. On the other hand, AuNP-related cardiotoxicity, in particular PEG-coated AuNPs, was quite limited. This demonstrates strong biocompatibility in terms of the cardiovascular system.
4.1.3. Complement Activation The complement system and its activation are components of the body’s innate immune system against foreign invaders. Thus, upon exposure to nanoparticles, complement activation may induce inflammatory responses but in some cases, these can become uncontrolled posing a serious threat [160]. Complement activation may also be responsible for some allergic reactions caused by different nanoparticle-based therapies, including cancer therapy. There are three general complement pathways: classical, lectin, and alternative pathways which converge in the formation of SC5b-9 complex as the final activation product prior to the destruction of cells [160]. In addition, several complement proteins such as C1q, C3b, and C4b function as opsonins and specifically-tagged nanoparticles for rapid clearance. However, these various processes and components of the complement system are highly affected by the surface coating of the nanoparticles.
Research on PEG-coated and citrate-capped AuNPs has indicated the differential effect on complement activation [161]. Ultimately citrate-capped AuNPs produced a size- dependent increase in the complement system end-product SC5b-9 in human serum, whereas the size-dependency was not present for PEG-coated AuNPs. Furthermore, PEG-coated AuNPs had a markedly reduced SC5b-9 level compared to citrate-capped, though it was still significantly increased compared with the control [161]. Another study with poly(2-methyl-2-oxazoline) (PMOXA) coated AuNPs demonstrated that this particular surface coating triggered complement activation to a greater extent only through the classical pathway [162]. The C1q mediated complement activation accelerated PMOXA opsonization and consequently, recognition by leukocytes and macrophages to a greater degree [162]. Greater clearance ability by the complement activation, without uncontrolled activation effects, would potentially decrease nanotoxicity as well. Complement activation effects are further summarized in Figure 3 as adapted from [160].
4.2. In Vivo Assessment
4.2.1. Gastrointestinal The gastrointestinal system is a complex, multi-organ system including the pharynx, esophagus, stomach, small and large intestines, rectum, liver, pancreas, and gallbladder. Nanoparticles have been employed in the treatment of various gastrointestinal cancers. The primary findings related to GI toxicity of nanoparticles depends on nanoparticle size and composition [163,164,165]. Specifically, studies on AuNPs have demonstrated that small particles (5 nm) preferentially produced pathological changes in the liver, whereas medium and large particles (20 nm and 50 nm) tended to target the spleen [163]. The toxic histopathological changes caused by the small AuNPs in the liver included steatosis, cytoplasmic degeneration, infiltration of inflammatory cells, Kupffer cells activation, and hemorrhage [163].
Further research on the cytotoxicity of AuNPs on different GI cancerous cell lines yielded variable results based on the composition of the conjugation or coating used. In a study by Huang et al. cell viability was greater than 90% in MGC803 gastric cancer cells after being exposed to AuNR@SiO2 targeted to folic acid, demonstrating that AuNPs by themselves were non-cytotoxic to MGC603 gastric cancer cells [164]. Similarly, another study by Li et al. incorporated chitosan AuNPs in esophageal cancer, which did not have any effect on benign human squamous esophageal epithelium cells or Barrett’s epithelium [165].
Another prominent type of nanoparticles used in biomedical applications are IONP or copper oxide (CuO) nanoparticles. The surface coating and size had a similar influence on overall toxicity patterns as demonstrated with AuNPs [166,167]. IONPs tend to accumulate in the liver and other RES organs. The iron products are recycled and then incorporated into pathways involved in hemoglobin, ferritin, and transferrin [166]. However, toxicity studies have indicated dose-dependent toxicity—high doses (2.5 mg/kg) could be potentially fatal as large aggregates formed quickly and rapid hemolysis can occur. When the anti-aggregating coating agent PEG was used with the IONPs, transient increases in the ALT enzyme was observed and there was much slower degradation and clearance of the PEG-coated IONPs [166]. Comparative toxicology research among SiO2, silver nanoparticles (AgNPs), and IONPs indicated the AgNPs caused a greater degree of GI systemic toxicity as demonstrated by increases in serum alkaline phosphatase and calcium, lymphocytic infiltration in the liver which was not observed in SiO2 and IONPs [167].
4.2.2. Renal Compared to the liver, the, kidneys tend to experience less nanoparticle, especially with AuNPs [168]. However, certain markers such as blood urea nitrogen, creatinine, protein, and globulin that have been found to be affected as a result of renal nanotoxicity. Studies have indicated a correlation between nanoparticle size and toxicity as 60 nm PEG-coated AuNPs demonstrated significant change in creatinine levels, indicating kidney toxicity, while sizes below 30 nm did not produce the same effect [169]. Within the kidneys, the proximal tubule epithelial cells were found to be the primary targets of nanotoxicity [170].
In one study, multiple cell lines from various organ systems, including the PK-15 cell line (epithelial porcine kidney) were studied for AuNP toxicity using dose-dependent and time-dependent measures [171]. At concentrations of 360 or 720 ng/mL, there was decreased cell growth within 24 h of addition. However, at concentrations below 360 ng/mL, the growth curves tended to shift back towards the control growth distribution and in some cases even exceed the baseline as time went on. This indicates that the toxic potential of gold nanoparticles at a reduced concentration are quite minimal and display some degree of reversibility as cell growth adapts and becomes resistant to change [171]. Histopathology of gold-related nanotoxicity indicated distorted glomeruli, mild necrosis, dilated tubules, and edema exudate. However, none of these findings were at the level of statistical significance as there was a great degree of variation among these minimal changes found in the renal system [171].
Comparison of renal toxicity effects of IONPs with AuNPs revealed similar trends: the distribution of iron in the kidneys was quite limited, with smaller particles being rapidly cleared in the urine [166]. At lower concentrations of IONPs, no significant change to the architecture of the kidneys occurs [172]. In addition, the coating further presented an effect on the renal toxicity potential. A study by Shukla et al. revealed that chitosan oligosaccharide-coated IONPs damaged kidney cell lines less, and showed less toxicity than bare IONPs in various cell lines, including Hek293 (human embryonic kidney), via MTT cell viability assay [173].
Overall, both AuNPs and IONPs appear to have limited cytotoxic impact on the renal system. However, modifications on the size, coating and concentration of the nanoparticles may enhance or decrease renal nanotoxicity as a result.
It is important to establish the interactions of engineered nanoparticles with their biological effects in order to realize their full potential in the clinic. Understanding the mechanisms of their toxicities will lead to a more rational design that can overcome some of the major hurdles in their translation. In vitro assessment involves the current studies on the molecular and cellular mechanisms associated with nanotoxicity in relation to cellular binding and persistence, complement activation, oxidative stress, inflammation, and DNA damage while in vivo assessment deals with systemic toxicities of some nanoparticles, both known and unknown, on the human body. The systemic toxicity of gold- and iron-based nanoparticles, the two nanoparticle systems frequently used in cancer diagnostics and therapy, are highlighted in this section.
4.1. In Vitro Assessment
4.1.1. Oxidative Stress, Inflammation, and DNA Damage Nanoparticles induce reactive oxygen species (ROS) production due to the presence of pro-oxidant functional groups on their surface, which causes an imbalance in the redox state of the cell [152,153]. Excess ROS production causes oxidative stress and activates pro-inflammatory responses such as decreased mitochondrial membrane potential and decreased antioxidant enzymatic activity, among other effects [152]. These responses then lead to DNA damage and apoptosis as the end point of the nanoparticle-induced toxicity to cells. Smaller nanoparticles more easily penetrate through cell membranes, their higher surface areas and surface reactivities present greater cytotoxicity due to increased ROS production [153].
In addition to increased production of ROS, nanoparticles induce cellular oxidative stress by depleting antioxidants that can combat mild oxidative stress [154]. Transcription and expression of antioxidant enzymes are regulated via nuclear factor Nrf2 induction. As the extent of oxidative stress increases, mitogen-activated protein kinase (MAPK) and NF-kB become activated as a pro-inflammatory responses [154]. However, excessive oxidative stress results in mitochondrial membrane damage and electron chain dysfunction, leading to DNA damage and eventually, apoptosis [154]. Silica or silicon dioxide nanoparticles (SiO2) were found to activate these oxidative stress-induced on human umbilical vein endothelial cells (HUVECs) [155]. The mRNA expression of Nrf2 and NF-kB was significantly upregulated [155], as illustrated in Figure 2.
Nanoparticles have been demonstrated to induce inflammatory responses in various cell types in different organ systems [152]. Some nanoparticles were recognized as pathogens by Toll-like receptors in the immune system, triggering increased production of inflammatory interleukins, chemokines, and adhesion molecules [155]. There is a strong link between inflammation and oxidative stress: inflammation potentially creates toxic by-products that promote the production of ROS while oxidative stress can result in the release proinflammation molecules, NF-kB and MAPK [152,154]. Research with SiO2 and TiO2 nanoparticles has indicated that inflammation through ROS generation can ultimately lead to changes in membrane permeability, leading to airway hypersensitivity reactions. Furthermore, the inflammatory and permeability effects have been proposed to extend beyond the lung and affect cardiovascular functioning as well [154]. Studies on specific molecular mechanisms associated with inflammation due to nanoparticle exposure are ongoing. Roy et al. found that inflammatory responses are linked to internalization of zinc oxide nanoparticles (ZnO) through endosome formation in macrophages, specifically by scavenger and caveolae pathways, in vitro [156]. The key inflammatory components studied included Cox-2 and iNOS expression, MAPKs, and cytokines such as IL-6, TNF-a, and IL-10. The results suggested that the inhibition of the caveolae internalization pathway reduces the expression of MAPKs [156].
DNA damage outcomes including, but not limited to DNA strand breaks, DNA protein cross-links, alkali-labile sites, and chromosomal aberrations, are observed upon oxidative stress induced by chronic exposure to nanoparticles [154]. Embryonic lung fibroblasts treated with AuNPs experienced DNA damage resulting to the formation of adducts with 8-hydroxydoxyguanosine and decreased expression of DNA repair and cell cycle checkpoint genes such as MAD2, cyclin B1, and cyclin B2 [152]. Regardless of the form of DNA damage, cells call upon repair mechanisms if the damage is reversible, or else transition into cell cycle arrest and apoptotic pathways. However, nanotoxicity has also been demonstrated to impact the DNA repair mechanisms themselves as seen in human embryonic lung fibroblasts exposed to AuNPs. In this case, DNA repair genes are downregulated and inability to repair the DNA damage would lead to apoptosis [152].
4.1.2. Cellular Binding and Persistence Cell-nanoparticle interaction studies allow for understanding of the cell adhesion, migration, and uptake pathways upon exposure to nanoparticles, both for drug-delivery efficacy and cytotoxic effects to healthy cells [157,158]. One key factor in the pathways of cell adhesion and migration is the doubling time of a particular cell line, as low doubling time indicates rapid proliferation and generally a higher migration efficiency [157]. The steps of cell migration include adhesion to the extracellular matrix, organization/disorganization of the actin cytoskeleton, membrane protrusion and retraction [157]. One particular study found that migration efficiency was dependent on the AuNP surface coating, but independent of size. The study proposed a potential mechanism in which cell adhesion to the nanoparticles occurs, then active migration and proliferation of cells and consequent cellular internalization sweeping out biocompatible nanoparticles in its way with non-biocompatible nanoparticles leading to cell toxicity [157].
Once cell adhesion and migration has occurred, the uptake component generally occurs through endocytosis, subdivided into phagocytosis and pinocytosis [158]. The key mechanisms of pinocytosis, which are involved in nanoparticle uptake include micropinocytosis, clathrin-mediated endocytosis, caveolae-mediated endocytosis, and clathrin- and caveolae-independent endocytosis [158]. Ultimately, most of their endocytic pathways lead to contents ending up in lysosomes for degradations [159]. However, in caveolae-mediated endocytosis, from endosomes, the contents form caveosomes, which are transported to the endoplasmic reticulum/Golgi apparatus, avoiding lysosomal degradation. This nuance in the uptake mechanisms was discovered as an important consideration for tailoring nanocarriers with drugs in cancer therapy [159].
Furthermore, the charge of the nanoparticles is critical in terms of cellular uptake and persistence [158]. Though previous studies have generalized the principle that positively charged nanoparticles interact more with cells compared to negatively charged ones due to the negative charge of the cell membrane, more current research indicates the greater complexity of this [158]. The protein corona is formed when nanoparticles are modified in a biological medium by various proteins that adsorb to the surface with different forces at play. Ultimately, studies have been mixed in terms of the effect of the protein corona on cellular uptake, with some indicating greater uptake with corona, while others demonstrated lesser uptake of nanoparticles. Thus, the rule of positive and negative charge on the nanoparticle surface cannot be simplified as such since there are multiple factors involving nanoparticle composition that affects cellular uptake [158].
Thanks Bruce! We will continue to use your help to crush the evil oppressors of humanity with the weight of their own LIES!!!! We appreciate your continued support - you are in a very small group of monthly supporters and we cant do this without you!
Oct 10, 2023·edited Oct 10, 2023Liked by Interest of Justice
The main protagonists have heaps of money and use the judicial process to make their opponents poor and bankrupt, rendering the cases against them as ineffective and never ending. Please support these people with whatever money you can spare. These are the modern day crusaders of our time who work tirelessly to make OUR LIVES SAFER and to bring about change for our children's futures.
'At a furious pace, the Netherlands is signing agreements in which it is giving up more and more of its own decision-making power, read sovereignty.
This has especially taken off during the Rutte cabinets. The pact was signed by outgoing Minister Kuipers, which allowed parliament to have little to no influence. The signing fell on the same day Klaus Swab visited Rutte in the Netherlands.
Following the awarding of the WEF FOOD Innovation hub to the Netherlands, in Wageningen, now follows this GLOBAL HEALTH HUB through which the Netherlands is attracting more and more globalist organizations to itself.
According to the ministry, THE GLOBAL HEALTH HUB NEDERLAND IS ESTABLISHED TO BE A FORerunner IN THE ORGANIZATION OF WORLD-WIDE HEALTH CARE.'
Note from me: Klaus Swab was in the Netherlands personally last week to be present at this signing event!
that is REALLY interesting the signing fell on the same day Klaus Swab visited Rutte in the Netherlands. Could the undue influence be any more obvious! Thanks for sharing. We need to learn more about this GLOBAL HEALTH AND FOOD HUB program??????? What the hell is this now? It never ends lol
Oct 3, 2023·edited Oct 3, 2023Liked by Interest of Justice
Super impressive. Super cool cartoon!!
You probably know about the case handled by Reiner Fuellmich in N.Zealand?
I wish I could send you 50 euro's a month (it isn't much but it is something...) but how do I do that without a credit card and Paypall? I ditched the damn things! Is there another way?
We have been so busy the past few days we just noticed this message Piki! Thank you so much for helping this mission. We are currently setting up the new websites and new donate portal and will for sure let you know! Yes there will be other ways to help than a credit card or PP - you are not the first to ask us to get our act together with a way to take checks and bank transfers, etc because donors don't want to be tracked supporting the one org going against global governance.. TOTALLY understood lol! working on it as we speak! Thanks for having our back on monthly - there are only a few monthly supporters and being able to rely on that from you would really help! We will make it happen and appreciate you! PS - the case in N Zealand is crazy because it has no police power, being the Maori tribe and all going against NATO.. Just saying lol. We pray to win here because its a real court that will be taken seriously and who can actually enforce the judgements! Lets do this!
Netherlands ready to merge into global medical-pharmaceutical complex
GLOBAL HEALTH PACT
Date: October 9, 2023
'The Dutch government has established a "Global Health Hub," preparing our country to become part of a global medical-pharmaceutical complex, driven by the World Health Organization (WHO) and the EU, that will determine Netherland's health policy. This includes the introduction of digital health passports and censorship of critical voices.
The Ministries of Health and Human Services and Foreign Affairs reported on Sept. 28 that they have signed "together with more than 20 parties" a Global Health Pact and established a Global Health Hub.'
I wish my lawyer worked near as hard as you! I'd win by a landslide. 🤗
Haha - we have to hire an attorney to go to trial and help structure and argue the main case too (why we are fundraising)- Its so aggravating to have to teach them and prod them and literally do ALL the work because in general they do the very MINIMUM... ughhhh. Whats your case in NY? Hope you win!!!!
Oh! Now, that's just sad!
No, I'm in the Eastern Midwest.
Thank-you for the good wishes!
Thanks for the update. I’m comfortable with slow as hell. Let them suffer longer.
Slow is always more thorough! We are praying for a fast win Nov 9 though to at least stop the shots locally in CR but hopefully globally. The laws here do allow the CR judge to have jurisdiction over the "superiors", in this case: WHO, FDA, CSC, EMA, etc. PRAY!!!!
Well this looks awesome, I will continue to pray everyday and ask God for protection, thank you for all you're doing.
What about the full list of ingredients for these poisonous injections?
It's easy to explain why they're toxic if they were exposed.
And preventing people from knowing what causes harm and, consequently, what can prevent it, what is an antidote to that toxicity, could save lives and prevent so many injuries...
Isn't that a crime???
Hiding the truth about the toxicity of ALL the ingredients?
Please see this post for information as it is the same technology:
https://outraged.substack.com/p/nanotechnology-is-toxic
There are two sides:
1. Contentious Administrative (the Nov 9 hearing) to prove its outside the limits of the law and VOID
2. Its criminal for the reasons you said and much more - Investigations are in process with parts concluded that are making their way to the correct court.
Hang tight, it's about to get interesting!
One more important thing:
It should be investigated whether TOXIC NANOTECHNOLOGY used to "prevent" "Covid" (in masks and tests and respiratory filters and many drugs, including Remdesivir) can cause the so-called Covid and can cause injury and death:
“A recent study of particular objects known as "nanotubes," revered for their extraordinary strength and electrical conductivity, demonstrated that such objects tend to clump within the lungs, causing suffocation.”
https://lawcat.berkeley.edu/record/1119251 Is Nanotechnology Prohibited by the Biological and Chemical Weapons Conventions
“Take the experience of researchers at DuPont, who are testing microscopic tubes of carbon, known as nanotubes, valued for their extraordinary strength and electrical conductivity.
When the researchers injected nanotubes into the lungs of rats in the summer of 2002, the animals unexpectedly began gasping for breath. Fifteen percent of them quickly died. ''It was the highest death rate we had ever seen,''
said David B. Warheit, the research leader, who began his career studying asbestos and has been testing the pulmonary effects of various chemicals for DuPont since 1984.
Early research has raised troubling issues. DuPont and others, for example, found evidence that the cells that break down foreign particles in rodent lungs have more trouble detecting and handling nanoparticles than larger particles that have long been studied by air pollution experts.
Lungs are not the only concern.
Research shows that nanoparticles deposited in the nose can make their way directly into the brain.
They can also change shape as they move from liquid solutions to the air, making it harder to draw general conclusions about their potential impact on living things. “
Carbon nanotubes are used in masks!
https://jnm.snmjournals.org/content/48/7/1039 Carbon Nanotubes: Potential Benefits and Risks of Nanotechnology in Nuclear Medicine
“Despite these potential benefits, the toxicity of CNTs (Carbon Nanotubes) is a major concern that needs to be more clearly understood and addressed. Pristine, water-insoluble CNTs have been found to be highly toxic in vitro to many different types of cells, including human keratinocytes, rat brain neuronal cells, human embryonic kidney cells, and human lung cancer cells. In addition, unmodified CNTs administered intratracheally to mice have been reported to induce the formation of lung granulomas. CNTs have also been shown to promote the aggregation of human platelets in vitro, and analogous carbon particulate matter found in the environment enhanced experimentally induced vascular thrombosis in rats.”
“[in 2015] Günter Oberdörster and co-authors published what is possibly the most comprehensive review of carbon nanotube toxicology studies to date. Focusing on inhalation of nanotubes, they document evidence of transient pulmonary inflammation, and rapid and persistent development of granulomatous lesions and interstitial fibrosis on exposure to single- and multiwalled carbon nanotubes in rodents. They cite data indicating that inhaled long and thin multiwalled carbon nanotubes can move to the lining surrounding the lungs and penetrate it, where they can potentially cause mesothelioma. Furthermore, the authors indicate that carbon nanotubes can act as a cancer promoter — with inhalation increasing the probability of developing lung cancer from exposure to other carcinogens.”
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4706753/ INHALATION EXPOSURE TO CARBON NANOTUBES (CNT) AND CARBON NANOFIBERS (CNF): METHODOLOGY AND DOSIMETRY - PMC (nih.gov)
Li et al. (2007a) compared the pulmonary response of mice exposed to purified MWCNT by IT instillation versus inhalation.
Intratracheal instillation produced inflammation and severe destruction of alveolar structures, while inhalation predominately resulted in moderate pathology consisting of alveolar wall thickening and cell proliferation but general alveolar structure was retained. This study demonstrated significant differences in the type and degree of pulmonary responses to MWCNT in mice between bolus-type IT instillation and inhalation, with higher doses deposited in lung by inhalation resulting in only moderate effects compared to severe lesions induced by instillation of lower doses. (!!!!!) - PCR "TEST"!!!! This is why it is done close to brain not close to lungs!!!!
https://outraged.substack.com/p/thats-what-covid-is
https://outraged.substack.com/p/bombshell-studies
https://outraged.substack.com/p/stop-quantum-tagging
There was no pandemic. No matter what they say or don’t say about the contents and technological design attributes of the jab, they’ll struggle here.
THEY'RE BOTH TRUE.
But there is a reason why SOME people die and get hurt by the so-called. "Covid." Why would someone inject/instill TOXIC nanotechnology/graphene/ethylene oxide through a so-called "PCR test"?
Why would someone add graphene and titanium oxide to make people inhale in so-called "masks"?
And why, the hell, add this nanotechnology to saline solution, "Remdesivir" (Veklury), "monoclonal antibodies", "C-19" injections (so-called "vaccines"), and all those others that are in the pipeline??? (Such as N-Plate - radiation poisoning "vaccine", RiVax - RW Malone "invention" against "castor poisoning", MarVax and other toxic injections against "Marburg", Ebola and so on???
So, although yes, there was no pandemic, toxic nanotechnology is killing and injuring people, causing "flu-like symptoms" and causing the FEAR of SUFFOCATION. (low saturation because of free and oxy radical, caused by oxidative stress/disbalance/nanotechnology toxicity...
CAN YOU PLESE ADDRESS THE TOXICITY OF NANOTECHNOLOGY IN YOUR STATEMENTS??? People listen to you. Can you take a look at this? As a drug maker you understand toxicology, right???
In short:
THEY KNOW WHAT DAMAGE these injections can cause!!!
They know what to use to mitigate the damage, although healthy people (and indeed no one) should get these injections – but they know how to prevent damage IF THESE NANOTECHNOLOGIES ARE INJECTED!!!
After all, ordinary people DO NOT EVEN KNOW WHAT they were injected with, so HOW SHOULD THEY KNOW what adverse effects these injections can cause and why people may die after these injections - and THEY HAVE NO IDEA how to save themselves!!!
5.
4.2.7. Immune In the immune system, nanoparticle immunotoxicity refers to the adverse effects such as complement activation-mediated pseudoallergy, hypersensitivity, immunosuppression, and inflammasome effects [199]. In general, most nanoparticles tend to accumulate in organs of the mononuclear phagocytic system such as the liver and spleen. The immune cells that in turn produce toxic effects upon nanoparticle exposure include monocytes, platelets, leukocytes, dendritic cells, and macrophages [199].
Coating AuNPs with polyethylene glycol (PEG) avoided immunotoxic responses [199,200]. One study compared the immunotoxic effects of coating AuNPs with PEG and chicken ovalbumin (OVA) [200]. No significant cytotoxicity to RAW264.7 macrophages was observed at AuNP concentrations of 20 µg/mL. However, the uptake capacity for the OVA-AuNPs was greater compared to PEG-coated AuNPs. Additionally, PEG-coated AuNPs did not induce a significant increase in TNF-a, IL-6 and IL-1B for AuNPs larger than 35 nm. In general, small nanoparticles, despite differences in surface coating, appeared to present greater immunotoxic effects compared to larger ones [200]. AuNP toxicity in murine and human lymphocytes has showed overall viability was only significantly reduced at 200 µg/mL, but not any concentration below this [201]. Another key immune cell, dendritic cells have been targeted as points of entry for immunotherapeutic agents using AuNPs. Research has indicated that dendritic cells have very low cytotoxicity upon exposure to different sizes and concentrations of AuNPs [202]. However, small AuNPs with size of 10 nm displayed weak apoptotic effects in dendritic cells compared to larger nanoparticles. In terms of surface coating, positive charged-polymer-coated AuNPs did have a significant cytotoxic effect [202]. This indicates the necessity for correct identification of surface chemistry when designing biocompatible nanoparticles for cancer therapy and diagnosis usage, among other medical uses.
On the other hand, the immunotoxicity profile of IONPs is different. Within macrophages, RAW264.7 macrophages treated with IONPs demonstrated an increase in oxidative stress and an increase in cell proliferation within 24 h [203]. Further, another study indicated that murine and human macrophage cell lines exposed to PEI-coated IONPs induced the activation of toll-like receptor 4 signaling and ROS production via different pathways that in turn further increased the overall activation of macrophages leading to pro-inflammatory effects [204]. With B and T lymphocytes, the effect of IONPs remains unclear as initial studies indicated no effect on function and cell viability. While the lack of significant cytotoxicity still holds true with current studies, changes have been observed in T-cell function including delays in proliferation rate [205]. However, it has been demonstrated that compared to control at 13 weeks post-IONP injection, the distribution of B cells decreased while T cells increased. In addition, the number of dendritic cells increased, though surface markers for antigen presentation such as CD40 were decreased [206]. This suppression of antigen presentation in dendritic cells was a common feature indicated in other studies [150]. Thus, with dendritic cells, cytotoxicity was not a significant feature, but functional impairment was present.
Overall, the effects of nanoparticle toxicity on the immune system are complex and multi-faceted as it involves a variety of different cell types across organ systems. In general, the current research suggests that AuNPs present limited toxicity to the cells of the immune system including macrophages, lymphocytes, and dendritic cells. However, the results with IONPs s are more mixed, with some functional impairment effects demonstrated, though the cytotoxic potential to immune cells remains low. Additionally, the surface coating and electrostatic charge of the nanoparticles play a key role in immunotoxicity profile and potential evasion of the immune system responses in targeted drug-delivery systems, an area for additional research.
etc.
It is also known that antioxidants can prevent oxidative imbalances and oxidative damage, which can cause death, blood clots, organ damage and other adverse effects.
4.
4.2.5. Pulmonary/Respiratory Nanoparticles are employed in different types of lung cancers as a method for targeted drug-delivery in therapy. Various research studies have examined the pulmonary toxicity associated with nanomedicine [187,188,189,190]. There seems to a degree of variability in the toxicity of AuNPs compared with other nanoparticle types, which appeared to be more consistent in their behavior [187,188]. A study by Avalos et al. compared the relative toxicity of silver- and gold-based nanoparticles on human pulmonary fibroblasts [187]. In general, the cytotoxic effect on the pulmonary fibroblasts was not size dependent for AuNPs, unlike some of the previous studies highlighted in sections on other organ systems. All sizes studied (30, 50, and 90 nm) demonstrated a reduction in cell mitochondrial activity and lactate dehydrogenase (LDH) leakage. Furthermore, in comparison to AgNPs, oxidative stress and production of ROS was greater with AuNPs in pulmonary fibroblast cells [188]. Another study examined three different human lung epithelial cell types (A549, BEAS-2b, and NHBE) for cytotoxic effects of AuNPs and AgNPs [188]. AuNPs were coated with either sodium citrate or chitosan, which created different surface charges on the particles. In general, A549 and BEAS-2B cells exhibited the least cytotoxic effects with an increase in LDH release only at the highest concentration of chitosan-coated AuNPs. However, NHBE cells were more affected in terms of cytotoxicity by AgNPs and AuNPs as measured by LDH release and membrane leakage [188].
Other researchers have investigated the cytotoxic effect of nanoparticles on key cells involved in the blood-air barrier in the pulmonary system [189,190]. In general, for different metal-organic frameworks, lung epithelial and alveolar macrophage cell lines were more adversely affected by lipid-coated nanoparticle systems [189]. Specifically, another study focused on an in vitro 3D lung model with three cell types of the epithelial tissue barrier: monolayer of alveolar cells, macrophages, and dendritic cells [190]. After initial exposure to AuNPs, there was no observable change in the cell morphology compared to control across the cell types. However, long-term effects are the current limitation of these research studies and remain unknown.
The general toxicity profile of IONPs in the pulmonary system is attributed to increased oxidative stress due to particle internalization, dissolution, release, and disruption of regular iron homeostasis [191]. In vivo studies have indicated that exposure to IONPs induces an elevated acute inflammatory response, which persists up to 28 days post-exposure. In addition, there was found to be an increase in the heat shock proteins and matrix metalloproteinases and evidence of progression to granulomas [191]. Additional research has focused on the toxic effects of metal nanoparticles following inhalation and intratracheal instillation as also are being used in the realm of targeted drug deliver in cancer [192,193]. After inhalational exposure of IONPs, there was a transient increase in acute total cell and neutrophil counts, pro-inflammatory chemokines, and oxidative stress in the initial time points. However, in the long-term there was no persistent inflammation at the end of the study demonstrating that IONPs had limited toxicity in the long-term [192].
Overall, while AuNPs appear to generally have adverse effects to the respiratory system, their significance in terms of long-term pulmonary cytotoxicity still varies depending on the type of AuNP and the particular lung cell type. Further studies are necessary to establish more generalized outlook on the effects of IONPs to various lung cells.
4.2.6. Reproductive Reproductive toxicity due to nanoparticles require further differentiation between their effects on male versus female reproductive systems. In general, nanoparticles have been demonstrated to cross the blood-testicle and blood-placenta barrier, pressing the importance of addressing reproductive toxicity [194]. Some studies have shown a decrease in sperm motility, albeit only at very high concentrations of AuNPs.
Female: Within the female reproductive system, nanoparticle accumulation tends to occur in the uteri and the ovaries [195]. In the case of AuNPs and IONPs, the greater accumulation within the uterus was observed for smaller nanoparticles. In addition, the negative impact on female sex hormones was largely seen in current studies with titanium nanoparticles (TiO2) as they increased expression of the Cyp17a1 gene which in turn increased estradiol, apoptotic-related genes, inflammatory and immune responses, among other effects [195]. Furthermore, some nanoparticles have shown to produce morphological changes in the follicles leading to a reduction in the mature oocytes present [196]. Ovarian toxicity was observed with long-term TiO2 nanoparticle, which caused a shapeless follicular antrum and irregular arrangement of cells, though, the results were inconclusive and not very general [196].
Male: In the male reproductive system, nanoparticle exposure tends to not only have an impact on the reproductive organs themselves, but also some potential effects on spermatogenesis and motility [195]. One particular study with AuNS found limited toxicity to the testes with no necrosis or histological disorganization within the germinal cells, spermatozoids, intertubular spaces, Leydig cells, and Sertoli cells in male mice [197]. On the other hand, additional research has indicated that exposure to zinc oxide nanoparticles (ZnO) presented with a reduction in testicular tissue and loss of cells in seminiferous tubules at an intraperitoneal dose of 250, 500, and 700 mg/kg/day [185]. Testicular toxicity due to ZnO has been established by several studies and some work has characterized the particular changes that manifest in the blood-testis barrier as well [196]. Mechanistically, ZnO have been suggested to trigger ROS, potentiate DNA lesions in germ cells, and downregulate the expression of gap junction proteins in the cell membrane [196]. Other researches have employed ligand-free and oligonucleotide-conjugated AuNPs to study toxic effects on spermatozoa specifically [198]. The findings indicated that sperm morphology and viability was generally not affected at any concentration [198].
In summary, within the male and female reproductive system, there is a limitation in current research on the cytotoxic potential of nanoparticles. The current studies seem to indicate that there are certain established adverse effects as those described in the spermatozoa and hormonal changes. However, the morphological changes to the reproductive organs and their long-term implications are highly dependent on the nanoparticle type, coating, and the cell type affected as with other organ systems [196].
3.
4.2.3. Nervous The nervous system is complex with many nuances that are not yet elucidated within the system itself, especially the brain. However, several research studies have examined aspects of nanotoxicity of AuNPs and IONPs, which will be the focus of this subsection.
Siddiqi et al. investigated changes in several biomarkers indicative of neurotoxicity upon injection of AuNPs in rat brains [174]. On one hand, there was a significance decrease in the enzyme glutathione peroxidase, which is an antioxidant in the brain that protects against oxidative damage. On the other hand, there were increases in markers of oxidative stress-derived DNA damage such as 8-hydroxydeoxyguanosine and heat shock protein 70 as well as apoptotic markers such as caspase-9. Furthermore, there was a significant increase in the neurotransmitters dopamine and serotonin, which demonstrated implications of AuNPs in potentiation of mood disturbances and chemical imbalances in the brain [174]. Other research has specifically studied the impact of AuNPs of different types on neural cells through the blood-retinal barrier [175]. In general, only small particles (less than 20 nm) were able to pass through the blood-retinal barrier and accumulate in the retinal layers. Furthermore, there was no toxicity demonstrated to neural cells such as retinal astrocytes, neurofilaments, and retinoblastoma cells in C57BL/6 mice [175].
The variability of toxicity effects on neural cells greatly depends upon the cell type and composition of the nanoparticles, which was demonstrated in other organ systems as well. For example, Joris et al. studied six different cell lines that included human and murine neural stem cells, human and mouse-derived progenitor cell lines, and human and murine neuroblastoma cell lines [176]. The study compared the relative toxicity effects of gold, iron oxide, and silver nanoparticles among these cell lines. Overall, it was found that AuNPs had the greatest degree of acute toxicity and IONPs having the lowest cytotoxic potential. In terms of the cell morphology, the C17.2 cell line was the only one with a reduction in cell area.
The cytotoxic potential of IONPs was further investigated on cultured neurons, astrocytes, and microglial cells. In rat cerebellar granular neurons, dimercaptosuccinate-coated IONPs accumulated within the cultured neurons by 1000-fold, but cellular integrity or viability was not adversely affected. This finding indicated the inherent potential of neurons to mediate oxidative stress effects through their antioxidant abilities [177]. While the finding with neurons was consistent with astrocytes, it did not hold true for microglial cells, which were rapidly damaged and displayed severe toxicity. At a mechanistic level, the rate of release of iron from internalized IONPs in microglial cells may be too rapidly transferred to lysosomes leading to toxic iron levels within the cells. Similarly, the cytotoxic potential of nanoparticles varies based on the particular mechanistic pathways associated with different nervous cell types to handle oxidative stress induced by the nanoparticles [176,177]. From a pathological perspective, accumulation of iron in the brain has been linked to neurodegenerative diseases such as aceruloplasminemia and neuroferritinopathy, as well as having the potential to play a role in Alzheimer’s and Parkinson’s diseases [178]. This research emphasizes the importance of finding the optimal amount of IONPs for therapeutic purposes in the nervous system.
4.2.4. Cardiovascular Cardiovascular nanomedicine has been employed in the diagnosis and treatment of cardiovascular diseases in addition to its role in cancer therapy [179]. One application of AuNPs in cancer therapy is the loading of these particles with doxorubicin (DOX, chemotherapy drug) for targeted drug delivery. The cardiotoxicity of these DOX-loaded and PEG-coated AuNPs have been studied in current pharmacological research [180]. Ultimately, DOX loaded onto AuNPs demonstrated no significant changes in cardiovascular function biomarkers such as serum lactate dehydrogenase (LDH) and creatinine kinase MB (CK-MB) levels compared to the free DOX. Similarly, another study further solidified these results in that DOX loaded on AuNPs did not create changes in CK-MB levels compared to baseline [181]. CK-MB is an enzyme in the myocardium that has served as the gold standard indicator of myocyte injury in many clinical and research settings [181].
There is limited research on the chronic cardiac toxicity of AuNPs, which was the aim of a study conducted by Yang et. al. on the effect of PEG-coated AuNPs at 2, 4, and 12-week time periods [182]. There was no significant decrease in the left ventricular ejection fraction across each time point for all sizes studied. Inflammatory mediators such as CD45+ and TNF-a indicated that chronic exposure to AuNPs did not spark inflammatory cell infiltration in the heart. Other studies that employed PEG-coated AuNPs have also concluded upon similar results that accumulation of AuNPs in the heart did not induce significant changes in cardiac hypertrophy, fibrosis or inflammation further demonstrating the strong biocompatibility of PEG-coated AuNPs in biomedical applications [183].
In the context of IONPs, cardiotoxic effects have been observed with IONPs in connection to myocardial damage due to iron accumulation. Unlike in the spleen, the macrophage clearance ability in the heart is limited, leading to longer-term accumulation not fully characterized in present research [184]. Particularly, some research has indicated that IV administration of IONPs resulted in a pro-coagulatory effect in vivo and in vitro while causing oxidative stress on the heart [185]. The effect of IONPs on different cardiac markers of oxidative stress in mice was also investigated. Data has indicated a significant increase of lipid peroxidation, reactive oxygen species, and superoxide dismutase in heart tissue compared with control groups [186].
Overall, there seems to be a certain degree of cardiotoxicity associated with IONPs, but the systemic effect of these microscopic changes is yet to fully be determined by current research. On the other hand, AuNP-related cardiotoxicity, in particular PEG-coated AuNPs, was quite limited. This demonstrates strong biocompatibility in terms of the cardiovascular system.
2.
4.1.3. Complement Activation The complement system and its activation are components of the body’s innate immune system against foreign invaders. Thus, upon exposure to nanoparticles, complement activation may induce inflammatory responses but in some cases, these can become uncontrolled posing a serious threat [160]. Complement activation may also be responsible for some allergic reactions caused by different nanoparticle-based therapies, including cancer therapy. There are three general complement pathways: classical, lectin, and alternative pathways which converge in the formation of SC5b-9 complex as the final activation product prior to the destruction of cells [160]. In addition, several complement proteins such as C1q, C3b, and C4b function as opsonins and specifically-tagged nanoparticles for rapid clearance. However, these various processes and components of the complement system are highly affected by the surface coating of the nanoparticles.
Research on PEG-coated and citrate-capped AuNPs has indicated the differential effect on complement activation [161]. Ultimately citrate-capped AuNPs produced a size- dependent increase in the complement system end-product SC5b-9 in human serum, whereas the size-dependency was not present for PEG-coated AuNPs. Furthermore, PEG-coated AuNPs had a markedly reduced SC5b-9 level compared to citrate-capped, though it was still significantly increased compared with the control [161]. Another study with poly(2-methyl-2-oxazoline) (PMOXA) coated AuNPs demonstrated that this particular surface coating triggered complement activation to a greater extent only through the classical pathway [162]. The C1q mediated complement activation accelerated PMOXA opsonization and consequently, recognition by leukocytes and macrophages to a greater degree [162]. Greater clearance ability by the complement activation, without uncontrolled activation effects, would potentially decrease nanotoxicity as well. Complement activation effects are further summarized in Figure 3 as adapted from [160].
4.2. In Vivo Assessment
4.2.1. Gastrointestinal The gastrointestinal system is a complex, multi-organ system including the pharynx, esophagus, stomach, small and large intestines, rectum, liver, pancreas, and gallbladder. Nanoparticles have been employed in the treatment of various gastrointestinal cancers. The primary findings related to GI toxicity of nanoparticles depends on nanoparticle size and composition [163,164,165]. Specifically, studies on AuNPs have demonstrated that small particles (5 nm) preferentially produced pathological changes in the liver, whereas medium and large particles (20 nm and 50 nm) tended to target the spleen [163]. The toxic histopathological changes caused by the small AuNPs in the liver included steatosis, cytoplasmic degeneration, infiltration of inflammatory cells, Kupffer cells activation, and hemorrhage [163].
Further research on the cytotoxicity of AuNPs on different GI cancerous cell lines yielded variable results based on the composition of the conjugation or coating used. In a study by Huang et al. cell viability was greater than 90% in MGC803 gastric cancer cells after being exposed to AuNR@SiO2 targeted to folic acid, demonstrating that AuNPs by themselves were non-cytotoxic to MGC603 gastric cancer cells [164]. Similarly, another study by Li et al. incorporated chitosan AuNPs in esophageal cancer, which did not have any effect on benign human squamous esophageal epithelium cells or Barrett’s epithelium [165].
Another prominent type of nanoparticles used in biomedical applications are IONP or copper oxide (CuO) nanoparticles. The surface coating and size had a similar influence on overall toxicity patterns as demonstrated with AuNPs [166,167]. IONPs tend to accumulate in the liver and other RES organs. The iron products are recycled and then incorporated into pathways involved in hemoglobin, ferritin, and transferrin [166]. However, toxicity studies have indicated dose-dependent toxicity—high doses (2.5 mg/kg) could be potentially fatal as large aggregates formed quickly and rapid hemolysis can occur. When the anti-aggregating coating agent PEG was used with the IONPs, transient increases in the ALT enzyme was observed and there was much slower degradation and clearance of the PEG-coated IONPs [166]. Comparative toxicology research among SiO2, silver nanoparticles (AgNPs), and IONPs indicated the AgNPs caused a greater degree of GI systemic toxicity as demonstrated by increases in serum alkaline phosphatase and calcium, lymphocytic infiltration in the liver which was not observed in SiO2 and IONPs [167].
4.2.2. Renal Compared to the liver, the, kidneys tend to experience less nanoparticle, especially with AuNPs [168]. However, certain markers such as blood urea nitrogen, creatinine, protein, and globulin that have been found to be affected as a result of renal nanotoxicity. Studies have indicated a correlation between nanoparticle size and toxicity as 60 nm PEG-coated AuNPs demonstrated significant change in creatinine levels, indicating kidney toxicity, while sizes below 30 nm did not produce the same effect [169]. Within the kidneys, the proximal tubule epithelial cells were found to be the primary targets of nanotoxicity [170].
In one study, multiple cell lines from various organ systems, including the PK-15 cell line (epithelial porcine kidney) were studied for AuNP toxicity using dose-dependent and time-dependent measures [171]. At concentrations of 360 or 720 ng/mL, there was decreased cell growth within 24 h of addition. However, at concentrations below 360 ng/mL, the growth curves tended to shift back towards the control growth distribution and in some cases even exceed the baseline as time went on. This indicates that the toxic potential of gold nanoparticles at a reduced concentration are quite minimal and display some degree of reversibility as cell growth adapts and becomes resistant to change [171]. Histopathology of gold-related nanotoxicity indicated distorted glomeruli, mild necrosis, dilated tubules, and edema exudate. However, none of these findings were at the level of statistical significance as there was a great degree of variation among these minimal changes found in the renal system [171].
Comparison of renal toxicity effects of IONPs with AuNPs revealed similar trends: the distribution of iron in the kidneys was quite limited, with smaller particles being rapidly cleared in the urine [166]. At lower concentrations of IONPs, no significant change to the architecture of the kidneys occurs [172]. In addition, the coating further presented an effect on the renal toxicity potential. A study by Shukla et al. revealed that chitosan oligosaccharide-coated IONPs damaged kidney cell lines less, and showed less toxicity than bare IONPs in various cell lines, including Hek293 (human embryonic kidney), via MTT cell viability assay [173].
Overall, both AuNPs and IONPs appear to have limited cytotoxic impact on the renal system. However, modifications on the size, coating and concentration of the nanoparticles may enhance or decrease renal nanotoxicity as a result.
1,
How do we know it is THE SAME TECHNOLOGY?
https://www.sciencedirect.com/science/article/abs/pii/S0013935123016742
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7692849/
4. Mechanisms of Nanoparticle Toxicity
It is important to establish the interactions of engineered nanoparticles with their biological effects in order to realize their full potential in the clinic. Understanding the mechanisms of their toxicities will lead to a more rational design that can overcome some of the major hurdles in their translation. In vitro assessment involves the current studies on the molecular and cellular mechanisms associated with nanotoxicity in relation to cellular binding and persistence, complement activation, oxidative stress, inflammation, and DNA damage while in vivo assessment deals with systemic toxicities of some nanoparticles, both known and unknown, on the human body. The systemic toxicity of gold- and iron-based nanoparticles, the two nanoparticle systems frequently used in cancer diagnostics and therapy, are highlighted in this section.
4.1. In Vitro Assessment
4.1.1. Oxidative Stress, Inflammation, and DNA Damage Nanoparticles induce reactive oxygen species (ROS) production due to the presence of pro-oxidant functional groups on their surface, which causes an imbalance in the redox state of the cell [152,153]. Excess ROS production causes oxidative stress and activates pro-inflammatory responses such as decreased mitochondrial membrane potential and decreased antioxidant enzymatic activity, among other effects [152]. These responses then lead to DNA damage and apoptosis as the end point of the nanoparticle-induced toxicity to cells. Smaller nanoparticles more easily penetrate through cell membranes, their higher surface areas and surface reactivities present greater cytotoxicity due to increased ROS production [153].
In addition to increased production of ROS, nanoparticles induce cellular oxidative stress by depleting antioxidants that can combat mild oxidative stress [154]. Transcription and expression of antioxidant enzymes are regulated via nuclear factor Nrf2 induction. As the extent of oxidative stress increases, mitogen-activated protein kinase (MAPK) and NF-kB become activated as a pro-inflammatory responses [154]. However, excessive oxidative stress results in mitochondrial membrane damage and electron chain dysfunction, leading to DNA damage and eventually, apoptosis [154]. Silica or silicon dioxide nanoparticles (SiO2) were found to activate these oxidative stress-induced on human umbilical vein endothelial cells (HUVECs) [155]. The mRNA expression of Nrf2 and NF-kB was significantly upregulated [155], as illustrated in Figure 2.
Nanoparticles have been demonstrated to induce inflammatory responses in various cell types in different organ systems [152]. Some nanoparticles were recognized as pathogens by Toll-like receptors in the immune system, triggering increased production of inflammatory interleukins, chemokines, and adhesion molecules [155]. There is a strong link between inflammation and oxidative stress: inflammation potentially creates toxic by-products that promote the production of ROS while oxidative stress can result in the release proinflammation molecules, NF-kB and MAPK [152,154]. Research with SiO2 and TiO2 nanoparticles has indicated that inflammation through ROS generation can ultimately lead to changes in membrane permeability, leading to airway hypersensitivity reactions. Furthermore, the inflammatory and permeability effects have been proposed to extend beyond the lung and affect cardiovascular functioning as well [154]. Studies on specific molecular mechanisms associated with inflammation due to nanoparticle exposure are ongoing. Roy et al. found that inflammatory responses are linked to internalization of zinc oxide nanoparticles (ZnO) through endosome formation in macrophages, specifically by scavenger and caveolae pathways, in vitro [156]. The key inflammatory components studied included Cox-2 and iNOS expression, MAPKs, and cytokines such as IL-6, TNF-a, and IL-10. The results suggested that the inhibition of the caveolae internalization pathway reduces the expression of MAPKs [156].
DNA damage outcomes including, but not limited to DNA strand breaks, DNA protein cross-links, alkali-labile sites, and chromosomal aberrations, are observed upon oxidative stress induced by chronic exposure to nanoparticles [154]. Embryonic lung fibroblasts treated with AuNPs experienced DNA damage resulting to the formation of adducts with 8-hydroxydoxyguanosine and decreased expression of DNA repair and cell cycle checkpoint genes such as MAD2, cyclin B1, and cyclin B2 [152]. Regardless of the form of DNA damage, cells call upon repair mechanisms if the damage is reversible, or else transition into cell cycle arrest and apoptotic pathways. However, nanotoxicity has also been demonstrated to impact the DNA repair mechanisms themselves as seen in human embryonic lung fibroblasts exposed to AuNPs. In this case, DNA repair genes are downregulated and inability to repair the DNA damage would lead to apoptosis [152].
4.1.2. Cellular Binding and Persistence Cell-nanoparticle interaction studies allow for understanding of the cell adhesion, migration, and uptake pathways upon exposure to nanoparticles, both for drug-delivery efficacy and cytotoxic effects to healthy cells [157,158]. One key factor in the pathways of cell adhesion and migration is the doubling time of a particular cell line, as low doubling time indicates rapid proliferation and generally a higher migration efficiency [157]. The steps of cell migration include adhesion to the extracellular matrix, organization/disorganization of the actin cytoskeleton, membrane protrusion and retraction [157]. One particular study found that migration efficiency was dependent on the AuNP surface coating, but independent of size. The study proposed a potential mechanism in which cell adhesion to the nanoparticles occurs, then active migration and proliferation of cells and consequent cellular internalization sweeping out biocompatible nanoparticles in its way with non-biocompatible nanoparticles leading to cell toxicity [157].
Once cell adhesion and migration has occurred, the uptake component generally occurs through endocytosis, subdivided into phagocytosis and pinocytosis [158]. The key mechanisms of pinocytosis, which are involved in nanoparticle uptake include micropinocytosis, clathrin-mediated endocytosis, caveolae-mediated endocytosis, and clathrin- and caveolae-independent endocytosis [158]. Ultimately, most of their endocytic pathways lead to contents ending up in lysosomes for degradations [159]. However, in caveolae-mediated endocytosis, from endosomes, the contents form caveosomes, which are transported to the endoplasmic reticulum/Golgi apparatus, avoiding lysosomal degradation. This nuance in the uptake mechanisms was discovered as an important consideration for tailoring nanocarriers with drugs in cancer therapy [159].
Furthermore, the charge of the nanoparticles is critical in terms of cellular uptake and persistence [158]. Though previous studies have generalized the principle that positively charged nanoparticles interact more with cells compared to negatively charged ones due to the negative charge of the cell membrane, more current research indicates the greater complexity of this [158]. The protein corona is formed when nanoparticles are modified in a biological medium by various proteins that adsorb to the surface with different forces at play. Ultimately, studies have been mixed in terms of the effect of the protein corona on cellular uptake, with some indicating greater uptake with corona, while others demonstrated lesser uptake of nanoparticles. Thus, the rule of positive and negative charge on the nanoparticle surface cannot be simplified as such since there are multiple factors involving nanoparticle composition that affects cellular uptake [158].
Keep the faith legal warriors. I will continue to donate monthly so you can crush these oppressors of humanity.
Bruce in NY.
Thanks Bruce! We will continue to use your help to crush the evil oppressors of humanity with the weight of their own LIES!!!! We appreciate your continued support - you are in a very small group of monthly supporters and we cant do this without you!
The main protagonists have heaps of money and use the judicial process to make their opponents poor and bankrupt, rendering the cases against them as ineffective and never ending. Please support these people with whatever money you can spare. These are the modern day crusaders of our time who work tirelessly to make OUR LIVES SAFER and to bring about change for our children's futures.
Every word you said is true! and thanks for drumming up the support!
IoJ, The WHO is transforming into something else : https://www.globalhealthhub.de/de/
and
"Netherlands Signs Global Health Pact"
'At a furious pace, the Netherlands is signing agreements in which it is giving up more and more of its own decision-making power, read sovereignty.
This has especially taken off during the Rutte cabinets. The pact was signed by outgoing Minister Kuipers, which allowed parliament to have little to no influence. The signing fell on the same day Klaus Swab visited Rutte in the Netherlands.
Following the awarding of the WEF FOOD Innovation hub to the Netherlands, in Wageningen, now follows this GLOBAL HEALTH HUB through which the Netherlands is attracting more and more globalist organizations to itself.
According to the ministry, THE GLOBAL HEALTH HUB NEDERLAND IS ESTABLISHED TO BE A FORerunner IN THE ORGANIZATION OF WORLD-WIDE HEALTH CARE.'
Note from me: Klaus Swab was in the Netherlands personally last week to be present at this signing event!
OMG look at this from the site you posted: “Global Mental Health” calls for a biopsychosocial approach and targeted information campaigns...
NOT GOOD. Yikes!
that is REALLY interesting the signing fell on the same day Klaus Swab visited Rutte in the Netherlands. Could the undue influence be any more obvious! Thanks for sharing. We need to learn more about this GLOBAL HEALTH AND FOOD HUB program??????? What the hell is this now? It never ends lol
Super impressive. Super cool cartoon!!
You probably know about the case handled by Reiner Fuellmich in N.Zealand?
I wish I could send you 50 euro's a month (it isn't much but it is something...) but how do I do that without a credit card and Paypall? I ditched the damn things! Is there another way?
We have been so busy the past few days we just noticed this message Piki! Thank you so much for helping this mission. We are currently setting up the new websites and new donate portal and will for sure let you know! Yes there will be other ways to help than a credit card or PP - you are not the first to ask us to get our act together with a way to take checks and bank transfers, etc because donors don't want to be tracked supporting the one org going against global governance.. TOTALLY understood lol! working on it as we speak! Thanks for having our back on monthly - there are only a few monthly supporters and being able to rely on that from you would really help! We will make it happen and appreciate you! PS - the case in N Zealand is crazy because it has no police power, being the Maori tribe and all going against NATO.. Just saying lol. We pray to win here because its a real court that will be taken seriously and who can actually enforce the judgements! Lets do this!
Thank you IoJ for all this info!!
Indeed. It will not end for a while.
Today in the Netherlands in De Andere Krant:
Netherlands ready to merge into global medical-pharmaceutical complex
GLOBAL HEALTH PACT
Date: October 9, 2023
'The Dutch government has established a "Global Health Hub," preparing our country to become part of a global medical-pharmaceutical complex, driven by the World Health Organization (WHO) and the EU, that will determine Netherland's health policy. This includes the introduction of digital health passports and censorship of critical voices.
The Ministries of Health and Human Services and Foreign Affairs reported on Sept. 28 that they have signed "together with more than 20 parties" a Global Health Pact and established a Global Health Hub.'