207 replies to this topic

[Kalliste]

I re-read this paper the other day and came upon this quote. Would it be correct to say that C60 works in the same way? Does it squench electrophiles of every major specie or only truly oxygen based ROS?

 

 

Here, we test the OS hypothesis of aging by studying the
effects on life-span of an artificial hydroxylamine scavenger
(IAC). IAC reacts with most—if not all—carbon, nitrogen
and oxygen reactive species of biological interest (including
peroxyl radicals (ROO*) and superoxide radical-anion
[O2*-] and was recently found to attenuate oxidative
diseases where OS has a pathophysiological role (17–19).
Unlike conventional antioxidants, IAC has an additional
action: upon quenching ROS, it becomes super-activated,
turning from a hydroxylamine to a nitroxide—an even more
potent and catalytic antioxidant (20,21; Scheme 1).
Hence, its antioxidant behavior is modulated by redox
homeostasis.

http://biomedgeronto...na.glu160.short

http://biomedgeronto...0.full.pdf html

 

To answer my own post I guess there is this paper. I'm not sure if it covers all the relevant ROS/RNS/RCS that exist though. I tried tracking the publications of these reserachers but googling chinese names is not an easy thing.

 

 

The scavenging of reactive oxygen species and the potential for cell protection
by functionalized fullerene materials
Jun-Jie Yin b,1, Fang Lao a,1, Peter P. Fu d, Wayne G. Wamer b, Yuliang Zhao a,c,*, Paul C. Wang f, Yang Qiu a,
Baoyun Sun c, Gengmei Xing c, Jinquan Dong c, Xing-Jie Liang a,**, Chunying Chen a,c,*
a Division of Nanomedicine and Nanobiology, National Center for Nanoscience and Technology of China, Beijing 100190, China
b Center for Food Safety and Applied Nutrition, Food and Drug Administration, College Park, MD 20740, USA
c Laboratory for Bio-Environmental Effects of Nanomaterials and Nanosafety, Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049, China
d Division of Biochemical Toxicology, National Center for Toxicological Research, Food and Drug Administration, Jefferson, AR 72079, USA
f Laboratory of Molecular Imaging, Department of Radiology, Howard University, Washington, DC 20060, USA

a b s t r a c t
We demonstrated that three different types of water-soluble fullerenes materials can intercept all of the
major physiologically relevant ROS. C60(C(COOH)2)2, C60(OH)22, and Gd@C82(OH)22 can protect cells
against H2O2-induced oxidative damage, stabilize the mitochondrial membrane potential and reduce
intracellular ROS production with the following relative potencies: Gd@C82(OH)22 C60(OH)22>
C60(C(COOH)2)2. Consistent with their cytoprotective abilities, these derivatives can scavenge the stable
2,2-diphenyl-1-picryhydrazyl radical (DPPH), and the reactive oxygen species (ROS) superoxide radical
anion (O2 ), singlet oxygen, and hydroxyl radical (HO), and can also efficiently inhibit lipid peroxidation
in vitro. The observed differences in free radical-scavenging capabilities support the hypothesis that both
chemical properties, such as surface chemistry induced differences in electron affinity, and physical
properties, such as degree of aggregation, influence the biological and biomedical activities of functionalized
fullerenes. This represents the first report that different types of fullerene derivatives can
scavenge all physiologically relevant ROS. The role of oxidative stress and damage in the etiology and
progression of many diseases suggests that these fullerene derivatives may be valuable in vivo cytoprotective
and therapeutic agents.

[bixbyte]

What if an insect like a roach ingested a quantity of C60 would the Fullerenes make them able to be immune from the toxicity of cyclic organic free radicals or insecticides? 

[Kalliste]

Here is an article written by the Porporato-fellow who published the study on MitoTEMPO, MitoQ and metastasis last year. I'm quoting the second half of the article which is of most interest to C60.

C60 seems to be good at going to Mitos, stopping superoxide and according to some testing,  good at stopping cancer.....

 

 

Superoxide is best known as a highly
reactive radical causing cellular damage
and death. This is actually true for high
levels of superoxide, which are associated
with extensive DNA damage, cytochrome
c release, and initiation of cell death programs.
At lower levels, however, superoxide
can initiate cellular signals promoting
adaptation to stress, thus increasing cellular
fitness. The concept that moderate levels
of mitochondria-derived reactive

oxygen species (mtROS) are beneficial for
physiology (mitohormesis) is supported
by experiments in C. elegans, in which a
transient increase in mitochondrial respiration
and mtROS production was shown
to improve resistance to stress and, eventually,
the lifespan of the worms.8 We propose
that the prometastatic concentration
range of mitochondrial superoxide lies
between these two levels.
Since the double Nobel Prize winner
Linus Pauling claimed that sky-high doses
of vitamin C could virtually prevent all
diseases, the use of general antioxidants as
food supplements or adjuvant treatments
with cancer has been popular among
patients, even though both Pauling and
his wife eventually died from cancer. This
theory is in no way supported by our
study. Indeed, compared to drugs like
mitoTEMPO and mitoQ, general antioxidants
indiscriminately target ROS at the
whole cell level, therefore affecting the
physiological balance between pro- and
anti-oxidants and potentially interfering
with conventional anticancer treatments.
They can also disrupt the tumor suppressor
ROS-p53 axis, as shown recently with
genetically modified mouse models of

BRAF-induced melanoma and KRASinduced
lung cancer: when treated with
the broad antioxidants N-acetylcysteine
and vitamin E, these mice displayed
increased tumor progression and metastasis.
9 On the contrary, inhibitors of mitochondrial
superoxide have been designed
to prevent ROS propagation and signaling
only from this particular organelle, without
affecting general redox homeostasis
and other subcellular sources of ROS. For
these reasons, we propose that these drugs
have the potential to become valuable
therapeutic tools for the prevention of
tumor metastasis, which must now be verified
with additional preclinical and clinical
tests.

 

http://www.tandfonli...548.2014.968043

 

 

[Kalliste]

 

Hydroxylated Fullerene: a potential anti-inflammatory and anti-oxidant agent for preventing mouse preterm birth

Tetsu Wakimoto, MD

,

Kaoru Uchida, PhD^$

,

Kazuya Mimura, MD

,

Takeshi Kanagawa, MD

,

Tzvetozar Roussev Mehandjiev, MD

,

Hisae Aoshima, PhD

,

Ken Kokubo, PhD

,

Nobuaki Mitsuda, MD, PhD

,

Yasuo Yoshioka, PhD

,

Yasuo Tsutsumi, PhD

,

Tadashi Kimura, MD, PhD

,

Itaru Yanagihara, MD, PhD

^$Present address: Faculty of Food and Nutrition, Kyushu Welfare University, Fukuoka, Japan

Received: March 3, 2015; Received in revised form: May 15, 2015; Accepted: July 13, 2015; Published Online: July 18, 2015

Publication stage: In Press Accepted Manuscript

DOI: http://dx.doi.org/10...jog.2015.07.017

Article Info

 

• Abstract

Abstract Objective

Intrauterine infection such as by Escherichia coli and Ureaplasma spp., induce placental inflammation and are one of the leading causes of preterm birth. Here we evaluated hydroxylated fullerene [C₆₀(OH)₄₄] for its in vitro anti-inflammatory and antioxidant effects against host cellular responses to the ureaplasma toll-like receptor 2 (TLR2) ligand, UPM-1. In addition, we investigated the preventative effects of C₆₀(OH)₄₄in vivo in a mouse preterm birth model that used UPM-1.

Methods

TLR2-overexpressing cell lines and the primary cultures of mouse peritoneal macrophages were pretreated with C₆₀(OH)₄₄. After UPM-1 addition to the cell lines, the activation of the nuclear factor κ-light chain-enhancer of activated B cells (NF- κB) signaling cascade and the production of reactive oxygen species were monitored. The levels of expression of inflammatory cytokines of interleukin (IL)-6, IL-1β, tumor necrosis factor (TNF)-α, and the production of reactive oxygen species were quantified after stimulation with UPM-1. The in vivo preventative effects of C₆₀(OH)₄₄ on mice preterm birth were evaluated by analyzing the preterm birth rates and fetal survival rates in the preterm birth mouse model with placental histological analyses.

Results

Pretreatment with C₆₀(OH)₄₄ significantly suppressed UPM-1-induced NF- κB activation and reactive oxygen species production in TLR2-overexpressing cell lines.

In the primary culture of mouse peritoneal macrophages, UPM-1-induced production of reactive oxygen species and the expression of inflammatory cytokines such as IL-6, IL-1β, and TNF-α were significantly reduced by pretreatment with C₆₀(OH)₄₄. In the UPM-1-induced preterm birth mouse model, the preterm birth rate decreased from 72.7% to 18.2% after an injection of C₆₀(OH)₄₄. Placental examinations of the group injected with C₆₀(OH)₄₄ reduced the damage of the spongiotrophoblast layer and reduced infiltration of neutrophils.

Conclusion

C₆₀(OH)₄₄ was effective as a preventative agent of preterm birth in mice.

 

[Kalliste]

 

Targeting ROS for therapy Suppressing ROS to inhibit proliferation

ROS contribute to mitogenic signaling, and thus decreasing intracellular ROS levels is an attractive method for inhibiting cancer growth. With this in mind, several large-scale studies have investigated whether supplementation with antioxidant vitamins, including β-carotene and vitamin A or vitamin E can reduce cancer risk in humans. Contrary to the expected result, supplementation increased the risk of cancer in both cases [96,97]. In agreement with these results, in genetic mouse models of K-Ras- or B-Raf-induced lung cancer, treatment with NAC or vitamin E markedly enhanced tumor growth and accelerated mortality [98]. These results show that the potential use of antioxidants for cancer therapy is complex and needs to be carefully validated before being applied. One possibility for the failure of these antioxidants as cancer treatments is their lack of specificity. Treatment of patients with general antioxidants may modulate many physiological processes that are relevant to cancer growth. For example, the immune system, an important modulator of cancer growth, has been shown to be sensitive to ROS levels [99]. Another possibility is that general antioxidants are differentially effective than targeted antioxidants. Mitochondrial-targeted versions of antioxidants have been shown to be potent inhibitors of cancer cell growth in vitro and in vivo[69,100]. Thus, further investigation needs to be considered to determine if targeted antioxidants are a viable method to treat cancer.

Another approach for inhibiting ROS is to decrease production. Decreasing mROS production necessarily involves inhibition of the ETC and thus may not be a practical due to toxicity inherent in inhibiting mitochondrial respiration. However, patients taking the antidiabetic drug metformin have recently been shown to have a reduced risk of cancer incidence and mortality [101]. Metformin has been shown to act as an inhibitor of complex I of the ETC [102,103]. We recently used a metformin insensitive complex I analog to confirm that the anticancer effect of metformin is primarily mediated by specific inhibition of complex I of cancer cells in vivo[104]. Interestingly, we also observed that treatment with metformin suppressed hypoxic activation of HIF1α, indicating that it may also decrease production of mROS under hypoxia. Whether this effect is important for the cancer suppressive effects of metformin requires further investigation. An alternative approach to decrease ROS production is by inhibiting NADPH oxidases. Indeed, loss of NADPH oxidase 4 has been shown to activate apoptosis in pancreatic cancer cells [105]. In addition, inhibitors of NADPH oxidase activity have been shown to have efficacy on mouse models of cancer in vivo[106,107].

Increasing ROS to selectively kill cancer cells

Considering that cancer cells have increased ROS levels, they may be selectively sensitive to the damaging effects of further increasing ROS. Increasing ROS production specifically in cancer cells is likely difficult to accomplish, although it is one proposed mechanism for how many current chemotherapeutics function [108]. Alternatively, since cancer cells frequently have increased expression of antioxidants to maintain homeostasis, a promising therapeutic approach is to inhibit antioxidants to expose cancer cells to endogenously produced ROS [109]. In support of this model, several small molecule screens identifying compounds that specifically inhibit growth of transformed cells have converged upon glutathione utilization [110-112]. In all cases, treatment with the identified small molecules decreased glutathione levels, increased ROS, and could be rescued by treatment with NAC. In addition, inhibition of antioxidant pathways has also been shown to be effective for inhibiting cancer growth. Genetic knockout of NRF2 inhibited disease progression in mouse models of pancreatic and lung cancer [31,32]. Inhibition of SOD1 by the small molecule ATN-224 was shown to cause ROS-dependent cancer cell death in vitro and decreased tumor burden in advanced K-Ras-driven lung cancers in vivo[113]. These recent examples provide further proof of principle that increasing ROS, whether by increasing production or inhibiting antioxidants, is a promising approach for targeting cancer cells (Figure 6). Further research is warranted to determine which components of the antioxidant pathway are selectively essential for tumor growth.

http://www.cancerand.../content/2/1/17

[Kalliste]

I posted this in the MitoQ thread but posting it here too since it's probably relevant to C60oo. If there is another Avian flu we should all stock up on some mito-antioxidants just in case

 

 

Here is an interesting paper from China on antioxidants and ROS-burst illnesses like Avian Flu, Ischemia. They mention a few interesting things.

 

 

Mitochondrion-Permeable Antioxidants to Treat ROS-Burst-Mediated Acute Diseases

http://downloads.hin.../aip/346827.pdf

 

 

Attached Files

•  netnytn.jpg   153.49KB   8 downloads

[Kalliste]

Todays harvest, I think one of these might be a repost but I'm a lazy bum.

 

 

 

Biomaterials. 2014 Nov;35(34):9269-79. doi: 10.1016/j.biomaterials.2014.07.030. Epub 2014 Aug 14.

The role of low levels of fullerene C60 nanocrystals on enhanced learning and memory of rats through persistent CaMKII activation.

Chen L¹, Miao Y¹, Chen L¹, Xu J¹, Wang X¹, Zhao H¹, Shen Y¹, Hu Y¹, Bian Y¹, Shen Y¹, Chen J¹, Zha Y¹, Wen LP², Wang M³.

Author information

Abstract

Engineered nanomaterials are known to exhibit diverse and sometimes unexpected biological effects. Fullerene nanoparticles have been reported to specifically bind to and elicit persistent activation of hippocampal Ca(2+)/calmodulin-dependent protein kinase II (CaMKII), a multimeric intracellular serine/threonine kinase central to Ca(2+) signal transduction and critical for synaptic plasticity, but the functional consequence of that modulation is unknown. Here we show that low doses of fullerene C60 nanocrystals (Nano C60), delivered through intrahippocampal infusion and without any obvious cytotoxicity in hippocampal neuronal cells, enhance the long-term potentiation (LTP) of rats. Intraperitoneal injection of 320 μg/kg of Nano C60, once daily for 10 days, also enhanced spatial memory of rats in addition to an increase of LTP. In parallel, both the IH and IP administration of Nano C60 increased the autonomous activity and the level of threonine 286 (T286) autophosphorylation of CaMKII, enhanced post-synaptic AMPA/NMDA ratio, and triggered time-dependent activation of ERK and CREB. Our results reveal a striking and highly unexpected ability of Nano C60 in positively modulating learning and memory, an effect that is most likely manifested through locking CaMKII in an active conformation, and may have significant implications for the potential therapeutic applications of fullerene C60, a classic engineered nanomaterial.

 

 

Moussa co-authored this one

 

 

In Silico Study of Spacer Arm Length Influence on Drug Vectorization by Fullerene C60
Haifa Khemir,1 Bahoueddine Tangour,1 and Fathi Moussa2

1Research Unity of Modeling in Fundamental Sciences and Didactics, Université de Tunis El Manar, IPEIEM, BP 254, El Manar 2, 2096 Tunis, Tunisia
2LETIAM, Lip(Sys)2, University of Paris Sud, IUT d’Orsay, Plateau de Moulon, 91400 Orsay, France

Received 12 January 2015; Revised 9 May 2015; Accepted 10 May 2015

Academic Editor: Miguel A. Correa-Duarte

Copyright © 2015 Haifa Khemir et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract

This work studies theoretically the effect of spacer arm lengths on the characteristics of a fullerene C60-based nanovector. The spacer arm is constituted of a carbon chain including a variable number of methylene groups (n = 2–11). To improve the ability of the fullerene carriage, two arms are presented simultaneously through a malonyl bridge. Then the evolution of selected physicochemical parameters is monitored as a function of the spacer arm length and the angle between the two arms. We show here that while the studied characteristics are almost independent of the spacer arm length or vary monotonically with it, the dipole moment and its orientation vary periodically with the parity of the number of carbon atoms. This periodicity is related to both modules and orientations of dipole moments of the spacer arms. In the field of chemical synthesis, these results highlight the importance of theoretical calculations for the optimization of operating conditions. In the field of drug discovery, they show that theoretical calculations of the chemical properties of a drug candidate can help predict its in vivo behaviour, notably its bioavailability and biodistribution, which are known to be tightly dependent of its polarity.

 

 

 

 

Physical consequences of the mitochondrial targeting of single-walled carbon nanotubes probed computationally

• V.A. Chistyakov^a,
• P.V. Zolotukhin^a,
• E.V. Prazdnova^a,
• I. Alperovich^{a, b, c, ,} ,
• A.V. Soldatov^b

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  doi:10.1016/j.physe.2015.03.005 Get rights and content

Highlights •

Interaction of single-walled carbon nanotube and H atom was simulated with DFT.

•

H atom was found to make a bond with the nanotube from outside of the carbon cage.

•

Possible implications and mechanism of intra-mitochondria potential alteration are discussed.

Abstract

Experiments by F. Zhou and coworkers (2010) [16] showed that mitochondria are the main target of the cellular accumulation of single-walled carbon nanotubes (SWCNTs). Our in silico experiments, based on geometrical optimization of the system consisting of SWCNT+proton within Density Functional Theory, revealed that protons can bind to the outer side of SWCNT so generating a positive charge. Calculation results allow one to propose the following mechanism of SWCNTs mitochondrial targeting. SWCNTs enter the space between inner and outer membranes of mitochondria, where the excess of protons has been formed by diffusion. In this compartment SWCNTs are loaded with protons and acquire positive charges distributed over their surface. Protonation of hydrophobic SWCNTs can also be carried out within the mitochondrial membrane through interaction with the protonated ubiquinone. Such “charge loaded” particles can be transferred as “Sculachev ions” through the inner membrane of the mitochondria due to the potential difference generated by the inner membrane. Physiological consequences of the described mechanism are discussed.

 

 

 

 

 

 

Study of Distribution and Biological Effects of Fullerene C₆₀ after Single and Multiple Intragastrical Administrations to Rats

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DOI: 10.1080/1536383X.2014.949695 Olga D. Hendrickson^a*, Olga V. Morozova^a, Anatoly V. Zherdev^a, Alexander I. Yaropolov^a, Sergey G. Klochkov^b, Sergey O. Bachurin^b & Boris B. Dzantiev^a

pages 658-668

Publishing models and article dates explained

 

Alert me

Abstract

In this study, male rats were intragastrically exposed to fullerene C₆₀ for 1-day and 30-day periods at daily doses of 2000 and 250 mg/kg of body weight, respectively. Fullerene was detected in organs and tissues by HPLC after the extraction from biosamples with toluene. No statistically significant differences in hematological and biochemical parameters of control and treated rats were found after single and multiple administrations. Throughout the observation periods no lethality was observed. At necropsy, no pathomorphological changes in internal organs were recorded. Fullerene was found in stomach, small intestine, liver, lungs, spleen, kidneys, and blood. The amounts of the detected fullerene in comparison to the administered doses are far smaller which is the evidence of its efficient excretion. Hence, it can be assumed that fullerene nanoparticles penetrate from the gastrointestinal tract of rats into the bloodstream and translocate into secondary organs with no pronounced toxic effect in experimental conditions studied.

http://www.tandfonli...95#.Vb4zXvkatLA

 

 

 

Nanocomposite Treatment Reduces Disease and Lethality in a Murine Model of Acute Graft-versus-Host Disease and Preserves Anti-Tumor Effects

Priscila T. T. Bernardes,^#1Bárbara M. Rezende,^#1Carolina B. Resende,¹Talles P. De Paula,²Alesandra C. Reis,¹William A. Gonçalves,¹Elias G. Vieira,³Maurício V. B. Pinheiro,³Danielle G. Souza,²Marina G. M. Castor,¹Mauro M. Teixeira,⁴ and Vanessa Pinho^1,*

Markus M. Heimesaat, Academic Editor

Author information ► Article notes ► Copyright and License information ►

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Abstract

Graft versus host disease (GVHD) is an immunological disorder triggered by bone marrow transplantation that affects several organs, including the gastrointestinal tract and liver. Fullerenes and their soluble forms, fullerols, are nanocomposites with a closed symmetrical structure with anti-inflammatory and anti-oxidant properties. The present study evaluated the effects of treatment with the fullerol (C60(OH)18-20) in the development and pathogenesis of GVHD in a murine model. Mice with experimental GVHD that were treated with the fullerol showed reduced clinical signs of disease and mortality compared with untreated mice. Treatment with the fullerol decreased the hepatic damage associated with reduced hepatic levels of reactive oxygen species, pro-inflammatory cytokines and chemokines (IFN-γ TNF-α, CCL2, CCL3 and CCL5) and reduced leukocyte accumulation. The amelioration of GVHD after treatment with the fullerol was also associated with reduced intestinal lesions and consequent bacterial translocation to the blood, liver and peritoneal cavity. Moreover, the fullerol treatment alleviated the GVHD while preserving effects of the graft against a leukemia cell line (GFP+P815). In summary, the fullerol was effective in reducing the GVHD inflammatory response in mice and may suggest novel ways to treat this disease.

 

http://www.ncbi.nlm....les/PMC4395348/

 

 

 

 

 

ACS Nano. 2015 May 26;9(5):4827-34. doi: 10.1021/nn506164s. Epub 2015 Apr 20.

Blocking the passage: C60 geometrically clogs K(+) channels.

Calvaresi M¹, Furini S², Domene C^3,4, Bottoni A¹, Zerbetto F¹.

Author information

Abstract

Classical molecular dynamics (MD) simulations combined with docking calculations, potential of mean force estimates with the umbrella sampling method, and molecular mechanic/Poisson-Boltzmann surface area (MM-PBSA) energy calculations reveal that C60 may block K(+) channels with two mechanisms: a low affinity blockage from the extracellular side, and an open-channel block from the intracellular side. The presence of a low affinity binding-site at the extracellular entrance of the channel is in agreement with the experimental results showing a fast and reversible block without use-dependence, from the extracellular compartment. Our simulation protocol suggests the existence of another binding site for C60 located in the channel cavity at the intracellular entrance of the selectivity filter. The escape barrier from this binding site is ∼21 kcal/mol making the corresponding kinetic rate of the order of minutes. The analysis of the change in solvent accessible surface area upon C60 binding shows that binding at this site is governed purely by shape complementarity, and that the molecular determinants of binding are conserved in the entire family of K(+) channels. The presence of this high-affinity binding site conserved among different K(+) channels may have serious implications for the toxicity of carbon nanomaterials.

Hrmm? Not sure what to make of this.

http://www.ncbi.nlm....pubmed/25873341

[Allen Walters]

I'm guessing c60/oo works on dogs. My son took the dog to the vet yesterday for some blood work due to excessive water consumption. Turns out she has a broken tooth. She turns 16 this fall and the vet told my son that if she didn't know how old the dog was and had to guess by the blood work, she would think she was a much younger dog. She acted shocked and made the comment that our dog is as healthy as a horse.

 

[Kalliste]

Thymic involution is dependent on redox problems. This is nice news for those of us who are younger and still have the time to save it via bioavailable effective antioxidants. I was 30 when I started C60 so that was probably too late, if C60 can help

 

 

Rapid aging of the thymus linked to decline in free radical defenses Date: August 6, 2015 Source: Cell Press Summary: A critical immune organ called the thymus shrinks rapidly with age, putting older individuals at greater risk for life-threatening infections. A new study reveals that thymus atrophy may stem from a decline in its ability to protect against DNA damage from free radicals.

The damage accelerates metabolic dysfunction in the organ, progressively reducing its production of pathogen-fighting T cells.

 

A critical immune organ called the thymus shrinks rapidly with age, putting older individuals at greater risk for life-threatening infections. A study published August 6 in Cell

Reports reveals that thymus atrophy may stem from a decline in its ability to protect against DNA damage from free radicals. The damage accelerates metabolic dysfunction in the organ, progressively reducing its production of pathogen-fighting T cells.

The findings suggest that common dietary antioxidants may slow thymus atrophy and could represent a promising treatment strategy for protecting older adults from infections.

 

"The thymus ages more rapidly than any other tissue in the body, diminishing the ability of older individuals to respond to new immunologic challenges, including evolving pathogens and the vaccines that may otherwise offer protection from them," says senior study author Howard Petrie of the Scripps Research Institute. "We provide, for the first time, a mechanistic link between antioxidants and normal immune function, opening new avenues for potential treatment strategies that could improve immune defenses in the aging population."

 

The thymus produces essential immune cells called T cells, which are continuously lost and must be replaced throughout life. But starting around the time of puberty, the thymus rapidly decreases in size and loses its capacity to produce enough new T cells. This loss is partially offset by the duplication of existing T cells, but the resulting population of cells becomes more and more biased toward memory T cells, which recognize pathogens from previous or ongoing infections. As a result, broad-spectrum immunity against new pathogens and protective immune responses elicited by new vaccines diminish with age.

The development of interventions to slow the progression of thymus atrophy has been limited by the lack of knowledge about the underlying mechanisms. The prevailing theory suggests that sex hormones play a key role, but this explanation does not account for the accelerated speed at which the thymus diminishes in size in comparison to other tissues. Moreover, the body of scientific evidence clearly indicates that other factors must be involved in age-related thymus atrophy.

To address this question, Petrie and first author Ann Griffith, currently at the University of Texas Health Science Center at San Antonio, developed a computational approach for analyzing the activity of genes in two major thymic cell types--stromal cells and lymphoid cells--in mouse tissues, which are very similar to human thymic tissues in terms of function and the properties of atrophy. They found that stromal cells were deficient in an antioxidant enzyme called catalase, resulting in the accumulation of free radical and metabolic damage.

To test whether catalase deficiency plays a causal role in thymus atrophy, the researchers performed genetic experiments to enhance catalase levels in mice. By 6 months of age, the size of the thymus of the genetically engineered mice was more than double that of normal mice. Moreover, mice that were treated with two common antioxidants from the time of weaning achieved nearly normal thymus size by 10 weeks of age.

Taken together, the findings provide support for the free-radical theory of aging, which proposes that reactive oxygen species such as hydrogen peroxide cause cellular damage that contributes to aging and a variety of age-related diseases. These toxic molecules, which form in cells as a natural byproduct of the metabolism of oxygen, have been linked to progressive atrophy in many organs and tissues as part of the normal aging process. However, these are generally slow, progressive processes that do not become apparent until late in life and often go mostly unnoticed.

"In the case of the thymus, atrophy is more rapid than other tissues, which we now show is a consequence of stromal catalase deficiency in the context of a highly metabolic environment designed to support the demands of T-cell proliferation," Petrie says. "Our studies show that, rather than an idiosyncratic relationship to sex steroids, thymic atrophy represents the widely recognized process of accumulated cellular damage resulting from lifelong exposure to the oxidative byproducts of aerobic metabolism."

In future studies, the researchers will investigate whether antioxidant supplementation improves the functioning of the thymus and the immune system during aging. If these studies provide support for this idea, then they could lead to the development of new clinical recommendations for the prevention or treatment of age-related thymus atrophy in humans.

 

Griffith et al. Metabolic damage and premature thymus aging caused by stromal catalase deficiency. Cell Reports, August 2015 DOI: 10.1016/j.celrep.2015.07.008

Edited by Cosmicalstorm, 07 August 2015 - 02:36 PM.

[bixbyte]

 

 

 

ACS Nano. 2015 May 26;9(5):4827-34. doi: 10.1021/nn506164s. Epub 2015 Apr 20.

Blocking the passage: C60 geometrically clogs K(+) channels.

Calvaresi M¹, Furini S², Domene C^3,4, Bottoni A¹, Zerbetto F¹.

Author information

Abstract

Classical molecular dynamics (MD) simulations combined with docking calculations, potential of mean force estimates with the umbrella sampling method, and molecular mechanic/Poisson-Boltzmann surface area (MM-PBSA) energy calculations reveal that C60 may block K(+) channels with two mechanisms: a low affinity blockage from the extracellular side, and an open-channel block from the intracellular side. The presence of a low affinity binding-site at the extracellular entrance of the channel is in agreement with the experimental results showing a fast and reversible block without use-dependence, from the extracellular compartment. Our simulation protocol suggests the existence of another binding site for C60 located in the channel cavity at the intracellular entrance of the selectivity filter. The escape barrier from this binding site is ∼21 kcal/mol making the corresponding kinetic rate of the order of minutes. The analysis of the change in solvent accessible surface area upon C60 binding shows that binding at this site is governed purely by shape complementarity, and that the molecular determinants of binding are conserved in the entire family of K(+) channels. The presence of this high-affinity binding site conserved among different K(+) channels may have serious implications for the toxicity of carbon nanomaterials.

Hrmm? Not sure what to make of this.

http://www.ncbi.nlm....pubmed/25873341

 

So this study leads us to believe that C60 might cause heart failure in some family types?

 

 

 

http://pubs.acs.org/....1021/nn506164s

 

"Experimentally it is observed that carbon nanoparticles inhibit EXP-2, KVS-1, human KCNQ1, Kv4.2, and human hERG potassium channels to different extents" ----> (I could not cut and paste this so I typed it in as correct as possible.)

  

Note wikipedia reference to hERG ---------------------------------->

 

hERG (the human Ether-à-go-go-Related Gene) is a gene (KCNH2) that codes for a protein known asK_v11.1, the alpha subunit of a potassium ion channel. This ion channel (sometimes simply denoted as 'hERG') is best known for its contribution to the electrical activity of the heart that coordinates the heart's beating (i.e., the hERG channel mediates the repolarizing I_Kr current in the cardiac action potential). When this channel's ability to conduct electrical current across the cell membrane is inhibited or compromised, either by application of drugs or by rare mutations in some families,^[1] it can result in a potentially fatal disorder called long QT syndrome; a number of clinically successful drugs in the market have had the tendency to inhibit hERG, and create a concomitant risk of sudden death, as a side-effect, which has made hERG inhibition an important antitarget that must be avoided during drug development.^[2] hERG has also been associated with modulating the functions of some cells of the nervous system^[3] and with establishing and maintaining cancer-like features in leukemic cells.^[4]

[bixbyte]

 

 

 

 

ACS Nano. 2015 May 26;9(5):4827-34. doi: 10.1021/nn506164s. Epub 2015 Apr 20.

Blocking the passage: C60 geometrically clogs K(+) channels.

Calvaresi M¹, Furini S², Domene C^3,4, Bottoni A¹, Zerbetto F¹.

Author information

Abstract

Classical molecular dynamics (MD) simulations combined with docking calculations, potential of mean force estimates with the umbrella sampling method, and molecular mechanic/Poisson-Boltzmann surface area (MM-PBSA) energy calculations reveal that C60 may block K(+) channels with two mechanisms: a low affinity blockage from the extracellular side, and an open-channel block from the intracellular side. The presence of a low affinity binding-site at the extracellular entrance of the channel is in agreement with the experimental results showing a fast and reversible block without use-dependence, from the extracellular compartment. Our simulation protocol suggests the existence of another binding site for C60 located in the channel cavity at the intracellular entrance of the selectivity filter. The escape barrier from this binding site is ∼21 kcal/mol making the corresponding kinetic rate of the order of minutes. The analysis of the change in solvent accessible surface area upon C60 binding shows that binding at this site is governed purely by shape complementarity, and that the molecular determinants of binding are conserved in the entire family of K(+) channels. The presence of this high-affinity binding site conserved among different K(+) channels may have serious implications for the toxicity of carbon nanomaterials.

Hrmm? Not sure what to make of this.

http://www.ncbi.nlm....pubmed/25873341

 

So this study leads us to believe that C60 might cause heart failure in some family types?

 

 

 

http://pubs.acs.org/....1021/nn506164s

 

"Experimentally it is observed that carbon nanoparticles inhibit EXP-2, KVS-1, human KCNQ1, Kv4.2, and human hERG potassium channels to different extents" ----> (I could not cut and paste this so I typed it in as correct as possible.)

  

Note wikipedia reference to hERG ---------------------------------->

 

hERG (the human Ether-à-go-go-Related Gene) is a gene (KCNH2) that codes for a protein known asK_v11.1, the alpha subunit of a potassium ion channel. This ion channel (sometimes simply denoted as 'hERG') is best known for its contribution to the electrical activity of the heart that coordinates the heart's beating (i.e., the hERG channel mediates the repolarizing I_Kr current in the cardiac action potential). When this channel's ability to conduct electrical current across the cell membrane is inhibited or compromised, either by application of drugs or by rare mutations in some families,^[1] it can result in a potentially fatal disorder called long QT syndrome; a number of clinically successful drugs in the market have had the tendency to inhibit hERG, and create a concomitant risk of sudden death, as a side-effect, which has made hERG inhibition an important antitarget that must be avoided during drug development.^[2] hERG has also been associated with modulating the functions of some cells of the nervous system^[3] and with establishing and maintaining cancer-like features in leukemic cells.^[4]

 

 

IMPORTANT READ THIS SCREEN SHOT:

 

 Screenshot from 2015-08-07 19:57:07.png   110.3KB   10 downloads

[Kalliste]

"Experimental tests are needed to confirm the existence of this intracellular binding site. If C60 is trapped in the cavity upon closure of the intracellular gate, it can result in a use-dependent open-channel block. This cumulative and indiscriminate blockage of K+ channels may have serious implications for the toxicity of carbon nanomaterials. "

It's a simulation, not scores of suddenly heartattacked test animals so I'm not gonna start screaming. I know they have given rats huge doses of c60 (2500mg/kg iirc) and nothing much happened. Perhaps the lipid thing with c60oo prevents this. Perhaps human users would need 500mg/kg to achieve it. Perhaps their simulation is wrong.
This kind of problem is a reminder that cycling and periodic dosing might be good until we know more.

[Kalliste]

They used corn oil.

 

 

J Toxicol Sci. 2012;37(2):353-61.

Sub-acute oral toxicity study with fullerene C60 in rats.

Takahashi M¹, Kato H, Doi Y, Hagiwara A, Hirata-Koizumi M, Ono A, Kubota R, Nishimura T, Hirose A.

Author information

Abstract

To obtain initial information on the possible repeated-dose oral toxicity of fullerene C60, Crl:CD(SD) rats were administered fullerene C60 by gavage once daily at 0 (vehicle: corn oil), 1, 10, 100, or 1,000 mg/kg/day for 29 days, followed by a 14-day recovery period. No deaths occurred in any groups, and there were no changes from controls in detailed clinical observations, body weights, and food consumption in any treatment groups. Moreover, no treatment-related histopathological changes were found in any organs examined at the end of the administration period and at the end of the recovery period. Blackish feces and black contents of the stomach and large intestine were observed in males and females at 1,000 mg/kg/day in the treatment group. There were no changes from controls in the liver and spleen weights at the end of the administration period, but those weights in males in the 1,000 mg/kg/day group increased at the end of the recovery period. Using liquid chromatography-tandem mass spectrometry, fullerene C60 were not detected in the liver, spleen or kidney at the end of the administration period and also at the end of the recovery period. In conclusion, the present study revealed no toxicological effects of fullerene C60; however, the slight increases in liver and spleen weights after the 14-day recovery period may be because of the influence of fullerene C60 oral administration. In the future, it will be necessary to conduct a long-term examination because the effects of fullerene C60 cannot be ruled out.

 

 

Edited by Cosmicalstorm, 08 August 2015 - 12:18 PM.

[niner]

They used corn oil.

 

J Toxicol Sci. 2012;37(2):353-61.

Sub-acute oral toxicity study with fullerene C60 in rats.

Takahashi M¹, Kato H, Doi Y, Hagiwara A, Hirata-Koizumi M, Ono A, Kubota R, Nishimura T, Hirose A.

Author information

Abstract

To obtain initial information on the possible repeated-dose oral toxicity of fullerene C60, Crl:CD(SD) rats were administered fullerene C60 by gavage once daily at 0 (vehicle: corn oil), 1, 10, 100, or 1,000 mg/kg/day for 29 days, followed by a 14-day recovery period. No deaths occurred in any groups, and there were no changes from controls in detailed clinical observations, body weights, and food consumption in any treatment groups. Moreover, no treatment-related histopathological changes were found in any organs examined at the end of the administration period and at the end of the recovery period. Blackish feces and black contents of the stomach and large intestine were observed in males and females at 1,000 mg/kg/day in the treatment group. There were no changes from controls in the liver and spleen weights at the end of the administration period, but those weights in males in the 1,000 mg/kg/day group increased at the end of the recovery period. Using liquid chromatography-tandem mass spectrometry, fullerene C60 were not detected in the liver, spleen or kidney at the end of the administration period and also at the end of the recovery period. In conclusion, the present study revealed no toxicological effects of fullerene C60; however, the slight increases in liver and spleen weights after the 14-day recovery period may be because of the influence of fullerene C60 oral administration. In the future, it will be necessary to conduct a long-term examination because the effects of fullerene C60 cannot be ruled out.

 

Hmm.  Takahashi et al.  came out in April 2012, so they didn't get a chance to see Baati before they submitted it.  In both cases, the animals were unharmed.   An important point from this paper is that solid c60 was excreted in feces, suggesting that if you have a few solid bits of c60 in your homemade c60oo, it's not likely to hurt you.  The combination of c60 and corn oil should create a fatty acid adduct similar to c60oo, if they allowed enough time for it to form.  Certainly in the 1000 mg/kg group, it would have been mostly a slurry of solid c60 in oil.

[niner]

"Experimental tests are needed to confirm the existence of this intracellular binding site. If C60 is trapped in the cavity upon closure of the intracellular gate, it can result in a use-dependent open-channel block. This cumulative and indiscriminate blockage of K+ channels may have serious implications for the toxicity of carbon nanomaterials. "

It's a simulation, not scores of suddenly heartattacked test animals so I'm not gonna start screaming. I know they have given rats huge doses of c60 (2500mg/kg iirc) and nothing much happened. Perhaps the lipid thing with c60oo prevents this. Perhaps human users would need 500mg/kg to achieve it. Perhaps their simulation is wrong.
This kind of problem is a reminder that cycling and periodic dosing might be good until we know more.

 

Exactly.  Simulations are low grade evidence, and it's easy to do them wrong.  They were looking at a single molecule of pristine c60, but you'll hardly ever see that in reality.  The closest thing would be Andrievsky's hydrated fullerenes, at least some of which may be single molecules, but they have a large shell of ordered water that would alter their behavior, possibly keeping them from docking in the channel.  In the case of c60oo, the attached fatty acid would be very likely to sterically prevent access to the channel. 

[bixbyte]

"Experimental tests are needed to confirm the existence of this intracellular binding site. If C60 is trapped in the cavity upon closure of the intracellular gate, it can result in a use-dependent open-channel block. This cumulative and indiscriminate blockage of K+ channels may have serious implications for the toxicity of carbon nanomaterials. "

It's a simulation, not scores of suddenly heartattacked test animals so I'm not gonna start screaming. I know they have given rats huge doses of c60 (2500mg/kg iirc) and nothing much happened. Perhaps the lipid thing with c60oo prevents this. Perhaps human users would need 500mg/kg to achieve it. Perhaps their simulation is wrong.
This kind of problem is a reminder that cycling and periodic dosing might be good until we know more.

 

It could be preemptive to have your MD do an EKG in office just to be certain you do not exhibit this symptom that could cause silent death.

The side effect in the study might not even be true.

But an ordinary EKG would pick up an abnormal Q-T wave delay.

That would be an easy way to evaluate a condition of dosing with C60.

I am fairly certain an EKG is covered by almost every medical insurance plan.  EKG, Big Deal, Free test noninvasive. 

If you do have these symptoms you would never have enough time to be able to report it.

[niner]

I'm not going to lose any sleep over a simulation using a compound that's a different molecule from the one I'm using, but for the record, I've had several EKGs performed in a competent cardiology practice since I've been using c60oo, with no abnormalities noted.

[Huckfinn]

Sorry if it's not the right place to post this...

I was about to order some (more) C60 to make my own mix.

I usually buy it from SES.

This time I received the message: "......the C60, 99.0% is no longer begin offered. Our lowest grade of C60 is 99.5%..."

 

Would that be fine you reckon?

 

Thanks!!

[niner]

I was about to order some (more) C60 to make my own mix.

I usually buy it from SES.

This time I received the message: "......the C60, 99.0% is no longer begin offered. Our lowest grade of C60 is 99.5%..."

 

Would that be fine you reckon?

 

 

99.5% is more pure than 99.0%, so it's closer to the 99.9+% that Baati used.  If you were ok with 99.0%, I don't understand why you would worry about 99.5%.  A lot of people use the higher purity grades, but to tell you the truth, I don't think it really matters all that much.  The impurity is mostly c70, which has similar antioxidant properties to c60.  A couple very interesting therapeutics from Luna Bioscience are c70-based.  These are anti-allergy and hair growth-promoting compounds.  One could speculate that the reports we've had of anti-allergy effects and hair growth from c60oo might be at least partially due to a c70 impurity, or might be more pronounced with c70oo.  If that's the case, then you might see more of those effects with the 99.0% product.

[pone11]

I'm guessing c60/oo works on dogs. My son took the dog to the vet yesterday for some blood work due to excessive water consumption. Turns out she has a broken tooth. She turns 16 this fall and the vet told my son that if she didn't know how old the dog was and had to guess by the blood work, she would think she was a much younger dog. She acted shocked and made the comment that our dog is as healthy as a horse.

 

What is the history of C60 use in that dog?   Number of years and what regimen?

[niner]

 

I'm guessing c60/oo works on dogs. My son took the dog to the vet yesterday for some blood work due to excessive water consumption. Turns out she has a broken tooth. She turns 16 this fall and the vet told my son that if she didn't know how old the dog was and had to guess by the blood work, she would think she was a much younger dog. She acted shocked and made the comment that our dog is as healthy as a horse.

 

What is the history of C60 use in that dog?   Number of years and what regimen?

 

I can't speak for Allen, but we have had a number of reports of old dogs perking up dramatically when given c60.  These were of the form "I gave my old dog c60oo and the next day he could [do something] that he hadn't done in years."

[Kalliste]

 

 

 

Carbon nanoparticles and oxidative stress: could an injection stop brain damage in minutes?

Lizanne G Nilewski​‌¹, William KA Sikkema​‌¹, Thomas A Kent​‌^2,3 & James M Tour​‌*^,1

*Author for correspondence: tour@rice.edu

Sections: ChooseChallenges in treatment o...Carbon nanoparticle-based...Applications to brain inj...Future perspectiveReferences

 

Challenges in treatment of oxidative stress ChooseTop of pageChallenges in treatment o... <<Carbon nanoparticle-based...Applications to brain inj...Future perspectiveReferences

Reactive oxygen species (ROS), which include superoxide, nitric oxide, hydroxyl radicals, nitrogen dioxide and peroxynitrite, play a crucial role in vital processes such as blood pressure regulation, cell migration, neurotransmission, immune regulation, microorganism defense and smooth muscle relaxation [1]. Under normal conditions in healthy systems, ROS are regulated by enzymes including superoxide dismutase (SOD) and catalase, as well as by small molecule natural antioxidants such as glutathione, ascorbic acid, coenzyme Q10, vitamin A and vitamin E. The overproduction of ROS leads to a condition known as oxidative stress, and at elevated concentrations, ROS readily cause oxidative damage to biomolecules including DNA, lipids, proteins and enzymes. Extreme levels of oxidative stress lead to extensive cellular damage, and these elevated levels of ROS often accompany acute injuries such as traumatic brain injury, inflammatory response and stroke as well as chronic diseases including cancer, cardiovascular dysfunction, rheumatoid arthritis, Parkinson's disease and Alzheimer's disease [1]. While natural antioxidants can regulate and alleviate oxidative stress to some degree, they have little effect on extremely elevated ROS levels, and this is due to their specific mechanisms of action. Many naturally occurring antioxidants and antioxidant enzymes act through chain reactions of radical transfers, often requiring cofactors, or need to act in concert with other antioxidants to be effective. For example, SOD converts two molecules of superoxide into oxygen and hydrogen peroxide, but it requires the presence of another enzyme, catalase, to complete the catalytic cycle. Additionally, as with SOD, many natural antioxidants can only quench one or two reactive species per molecule before requiring regeneration. Thus, these natural antioxidants can be easily overwhelmed by the highly elevated levels of ROS during disease or traumatic injury, and the effects are exacerbated when the trauma is accompanied by hemorrhagic shock. Overall, these limitations of natural antioxidants explain why most studies of antioxidant therapies in a clinical setting show little to no benefit in disease or injury treatment [2]. The benefit there is from antioxidant therapy is mostly apparent in models where the antioxidant is administered before the injury takes place, and few have been shown to have any significant affect postinjury. There is therefore a distinct need to develop synthetic antioxidant treatments that renormalize elevated levels of oxidative stress, and especially therapies that can be administered after an injury has taken place. An optimal antioxidant would be able to regenerate itself without needing a cascade of other enzymes or small molecules to complete the cycle, rapidly react with multiple ROS, be able to quench reactive species without creating potentially harmful byproducts, not react with helpful radicals, such as the vasodilator nitric oxide, show efficacy when administered postinjury, and be nontoxic and readily cleared post elevated levels of ROS. If such an antioxidant could be realized, it could act orthogonally to the cell's natural antioxidant system and thereby mitigate the ROS independently, allowing it to be effective when the cell's antioxidant system is compromised.

Carbon nanoparticle-based antioxidants ChooseTop of pageChallenges in treatment o...Carbon nanoparticle-based... <<Applications to brain inj...Future perspectiveReferences

Our team has discovered nontoxic carbon nanoparticle-based antioxidants that have reactivity profiles toward ROS that fit with the optimal profile. The nanoparticles are called PEG-functionalized hydrophilic carbon clusters (PEG-HCCs). These are 3 × 40 nm highly edge-oxidized PEGylated fragments of pi-conjugated monoatomic layer graphene-like domains [3]. The PEG-HCCs are nontoxic, and they have a half-life of 2 to 3 h in vivo. Encouragingly, these PEG-HCC carbon nanoparticles show antioxidant activity that has not been realized for any enzymatic system or small molecule therapy.

Originally, PEG-HCCs were used as passive drug delivery nanovectors, where the hydrophobic network of sp² carbons acts as a core that can physisorb similarly hydrophobic drugs and targeting antibodies. The oxidation on the edges of the HCCs and the PEG moieties allows the entire complex to be water or phosphate-buffered saline soluble, permitting intravenous delivery of the drug/PEG-HCC combination where the drugs are physisorbed to the conjugated domains. The use of paclitaxel/PEG-HCCs has been shown to be as effective as the standard drug formulation Taxol™ [4], the latter requiring patient premedication of high-dose corticosteroids to avoid clinical acute hypersensitivity to the surfactant Cremophor EL. The Cremophor EL is used to maintain water solubility of the otherwise insoluble paclitaxel. The PEG-HCCs can also be used in an antibody/drug/PEG-HCC formulation where both the drug and the antibody are physisorbed to the HCC core. Using an EGFR antibody physisorbed to the surface of the PEG-HCCs, we are able to selectively target an EGFR+ tumor over an EGFR- tumor in a single mouse [5]. We have also found that using short peptides that are covalently attached to the PEG termini, drug/peptide–PEG-HCC combinations are effective at treating several cancer types [6]. Thus, PEG-HCCs can be targeted through noncovalent antibody delivery, or through covalently bound peptide recognition schemes, thereby minimizing nanoparticle dosages.

Extending beyond the passive drug delivery applications of the PEG-HCCs, they can act as powerful active antioxidants due to the extended pi-conjugated domains of the HCCs. Like some fullerene derivatives which also contain extended pi-conjugation [7], PEG-HCCs are antioxidants that quench ROS, such as superoxide and hydroxyl radicals, in cell-free systems [3], in vitro and in vivo [4–5,8]. However, PEG-HCCs act much more quickly than any fullerene-based system. The turnover rate of superoxide to oxygen by PEG-HCCs was measured to be >20,000 s⁻¹, which is higher than most single-active-site enzymes, suggesting that the PEG-HCCs have multiple active sites [9]. This allows the PEG-HCCs to be effective even when administered after the traumatic event, making them clinically unique [10].

Applications to brain injury ChooseTop of pageChallenges in treatment o...Carbon nanoparticle-based...Applications to brain inj... <<Future perspectiveReferences

In cases of extreme oxidative stress in the brain, for example, in conditions of stroke or traumatic brain injury accompanied by hemorrhagic shock, the rapid kinetics of the PEG-HCCs allows them to act quickly to stop brain damage. In addition to the elevated levels of superoxide produced during the initial injury, a second wave of superoxide is produced by cells as the injury is being treated by reperfusion of blood. One contribution to this wave is the buildup of hypoxanthine during hypoxia which suddenly reacts via xanthine oxidase with the oxygen in reperfused blood to produce superoxide [11]. This second spike of superoxide can be addressed clinically by administration of the high-capacity antioxidants PEG-HCCs directly before reperfusion [12]. Indeed, in a rat model of traumatic brain injury accompanied by hemorrhagic shock, measurements taken after PEG-HCC treatment and blood reperfusion showed that animals treated with PEG-HCCs had normal levels of superoxide in the brain and vasculature, while those not treated with PEG-HCCs had as much as double the levels of superoxide. Additionally, the PEG-HCCs did not react with the essential nitric oxide. Rather, it was found that PEG-HCCs normalized nitric oxide levels in the treatment group, while in the controls, the nitric oxide levels were significantly lowered, presumably due to reaction between nitric oxide and the excess superoxide to form the highly deleterious peroxynitrite [10]. Preliminarily, PEG-HCCs have been extended to stroke models where the PEG-HCCs are delivered before clearance of the clot, potentially mitigating superoxide buildup during blood reperfusion to the brain.

Future perspective ChooseTop of pageChallenges in treatment o...Carbon nanoparticle-based...Applications to brain inj...Future perspective <<References

Injury and disease due to oxidative stress extend far past the brain. Cancer becomes more metastatic under oxidative stress conditions [13], inflammation and superoxide are intimately coupled [11,14], and superoxide plays a role in numerous chronic disorders such as diabetes, Parkinson's and Alzheimer's diseases [1,15]. PEG-HCCs have been shown to carry out therapeutic functions in preclinical models that have been unattainable from enzymes or small molecule treatment, and this could be a harbinger for the use of active carbon nanoparticle-based therapies. Therefore, PEG-HCCs could soon enjoy a prominent place in the treatment of acute injuries and chronic disorders.

 

 

I emailed the PEG-HCC people a couple of months ago and queried them about C60, they never replied but they make a quiet reference to C60 in this recent paper.

 

[Turnbuckle]

Here is an interesting paper discussing the solubility of C60 and C70 in food oils and the way it changes color from purple to red. It's interesting that C60 goes through a three stage solution process whereas C70 has only one stage and goes into solution much faster. This is rationalized as a result of the greater free volume in C70 crystals, but that is not entirely convincing. Regarding C60--

 

 

At this stage, attaching fatty acids to the outer surface of fullerene molecules occurs. Reaction is accompanied by the change of solution color – now it can be defined as “ripe cherry”, the completion of the reaction leads to a further darkening of the solution.

 

 

 

 

[Kalliste]

Mitoflashes may not exist after all, there is some confusion about the probes used for those trials anyway:

 

 

The ‘mitoflash’ probe cpYFP does not respond to superoxide

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Arising from E.-Z. Shen et al. Nature 508, 128–132 (2014); doi:10.1038/nature13012

Ageing and lifespan of organisms are determined by complicated interactions between their genetics and the environment, but the cellular mechanisms remain controversial; several studies suggest that cellular energy metabolism and free radical dynamics affect lifespan, implicating mitochondrial function. Recently, Shen et al.¹ provided apparent mechanistic insight by reporting that mitochondrial oscillations of ‘free radical production’, called ‘mitoflashes’, in the pharynx of three-day old Caenorhabditis elegans correlated inversely with lifespan. The interpretation of mitoflashes as ‘bursts of superoxide radicals’ assumes that circularly permuted yellow fluorescent protein (cpYFP) is a reliable indicator of mitochondrial superoxide², but this interpretation has been criticized because experiments and theoretical considerations both show that changes in cpYFP fluorescence are due to alterations in pH, not superoxide^{3, 4, 5, 6, 7}. Here we show that purified cpYFP is completely unresponsive to superoxide, and that mitoflashes do not reflect superoxide generation or provide a link between mitochondrial free radical dynamics and lifespan. There is a Reply to this Brief Communication Arising by Cheng, H. et al. Nature 514, http://dx.doi.org/10.1038/nature13859 (2014).

 

Statines and mitochondrial damage, interesting considering what some statin users have reported about C60

 

Physiology 2015 (Cardiff, UK) (2015) Proc Physiol Soc 34, C21

Oral Communications

Statin induced myopathy: a role for NO and ROS in enhanced sarcoplasmic reticulum Ca2+ leak

S. Lotteau1, D. MacDougall1, D. S. Steele1, S. Calaghan1

1. School of Biomedical Sciences, University of Leeds, Leeds, United Kingdom.

Medline articles by:

• Lotteau, S
• MacDougall, D
• Steele, DS
• Calaghan, S

 

 

Myopathy is the most common side effect of statins and its incidence is under-reported by randomised controlled clinical trials (1.5-5%) because of exclusion of susceptible individuals 1. The need to understand the mechanism of statin-induced myopathy is increasing as the cardiovascular risk threshold for statin prescription is reduced across the globe. We have previously shown that statin treatment increases the frequency and duration of Ca2+ sparks/embers in intact fast skeletal muscle fibres from the rat, consistent with increased sarcoplasmic reticulum (SR) Ca2+ leak (a common myopathic mechanism) 2. Here we investigate a role for nitric oxide (NO) and reactive oxygen species (ROS) in this increased leak. Male Wistar rats were treated with simvastatin 40 mg/kg/day by oral gavage over a 4 week period. Intact (non-permeabilised) type II skeletal muscle fibres from flexor digitorum brevis (FDB) were isolated by collagenase digestion and loaded with fluorescent dyes (Fluo-4AM, DAF-2 or JC-1). Data are given as mean ± S.E.M. of 17-50 cells from 5-11 rats, and compared with the Student's t-test or 2 way ANOVA. Inhibition of NOS with L-NAME (1 mM) had a greater impact (P<0.05) on NO (indexed with DAF-2) in statin fibres compared with controls, consistent with increased NOS activity with statin treatment. Inhibition of NOS reduced spark frequency by 60% (P<0.01) in fibres from statin-treated animals, but was without effect in controls, suggesting a role for NO in increased leak. Expression of caveolin 1 and 3 (normalised to GAPDH) was reduced by 67% (P<0.05) and 24% (P>0.05) in gastrocnemius (GAS) muscle from statin treated animals; this could contribute to increased NOS activity as caveolins are the main constitutive inhibitors of nNOS and eNOS. There was also evidence of increased ROS production with statin treatment. Mitochondrial membrane potential (indexed with JC-1) was reduced in statin fibres compared with controls (1.1 ± 0.1 vs. 1.7 ± 0.2 AU; P<0.05). Uncoupling of the mitochondrial respiratory chain increases ROS production. We also saw a trend for a decrease in the ratio of reduced:oxidised gluthatione (GSH:GSSG) in GAS from statin-treated animals compared with controls (12 ± 1 vs. 14 ± 1; P>0.05). Importantly, the superoxide dismutase mimetic MnTMPyP (0.1 mM) and the mitochondrial ROS scavenger Mitotempo (25 µM) both significantly reduced (P<0.05) spark frequency in statin fibres but were without effect in controls. Together these data show that increased SR Ca2+ leak seen in intact muscle fibres from statin-treated rats is linked to an increase in NO and ROS. We propose that caveolin-regulated NO and Ca2+-dependent mitochondrial ROS production modify the RyR to effect this leak. Defining the cellular processes that underlie statin induced myopathy is the first step in the development of co-therapies to improve statin compliance.

 

 

 

[Allen Walters]

 

I'm guessing c60/oo works on dogs. My son took the dog to the vet yesterday for some blood work due to excessive water consumption. Turns out she has a broken tooth. She turns 16 this fall and the vet told my son that if she didn't know how old the dog was and had to guess by the blood work, she would think she was a much younger dog. She acted shocked and made the comment that our dog is as healthy as a horse.

 

What is the history of C60 use in that dog?   Number of years and what regimen?

 

 

I don't know the exact dose she gets. I work out and eat between 2-2.5 lbs of chicken a day, I use a big jar of chicken gravy with 1 Tbs of c60/oo mixed in and divide it up into 3 meals. The last meal of the day I eat all but a few bites of the chicken and give the rest to her. She isn't getting much that's for sure. I contacted the vet to ask her about the blood work and she seemed very interested in the c60. She told me she had never seen enzyme levels that good in a dog that old. I've been giving her the c60 for at least a year now.

 

Edited by Allen Walters, 14 August 2015 - 05:26 PM.

[Huckfinn]

 

I was about to order some (more) C60 to make my own mix.

I usually buy it from SES.

This time I received the message: "......the C60, 99.0% is no longer begin offered. Our lowest grade of C60 is 99.5%..."

 

Would that be fine you reckon?

 

 

99.5% is more pure than 99.0%, so it's closer to the 99.9+% that Baati used.  If you were ok with 99.0%, I don't understand why you would worry about 99.5%.  A lot of people use the higher purity grades, but to tell you the truth, I don't think it really matters all that much.  The impurity is mostly c70, which has similar antioxidant properties to c60.  A couple very interesting therapeutics from Luna Bioscience are c70-based.  These are anti-allergy and hair growth-promoting compounds.  One could speculate that the reports we've had of anti-allergy effects and hair growth from c60oo might be at least partially due to a c70 impurity, or might be more pronounced with c70oo.  If that's the case, then you might see more of those effects with the 99.0% product.

 

Thanks Niner.

The reason I was asking was basically because at SES they now charge just over 160$ for 5gr of C60 at 99.5% as opposed to the approx. 130$ I was paying them before for just 1gr at 99.0%...

Bizarre, isn't it?

[Turnbuckle]

 

 

I was about to order some (more) C60 to make my own mix.

I usually buy it from SES.

This time I received the message: "......the C60, 99.0% is no longer begin offered. Our lowest grade of C60 is 99.5%..."

 

Would that be fine you reckon?

 

 

99.5% is more pure than 99.0%, so it's closer to the 99.9+% that Baati used.  If you were ok with 99.0%, I don't understand why you would worry about 99.5%.  A lot of people use the higher purity grades, but to tell you the truth, I don't think it really matters all that much.  The impurity is mostly c70, which has similar antioxidant properties to c60.  A couple very interesting therapeutics from Luna Bioscience are c70-based.  These are anti-allergy and hair growth-promoting compounds.  One could speculate that the reports we've had of anti-allergy effects and hair growth from c60oo might be at least partially due to a c70 impurity, or might be more pronounced with c70oo.  If that's the case, then you might see more of those effects with the 99.0% product.

 

Thanks Niner.

The reason I was asking was basically because at SES they now charge just over 160$ for 5gr of C60 at 99.5% as opposed to the approx. 130$ I was paying them before for just 1gr at 99.0%...

Bizarre, isn't it?

 

 

You likely remember that wrong. Likely you bought 99.9% before.

[Turnbuckle]

I posted this in the supplier thread, but it's more appropriate here. On the subject of the oxidation of C60 to form an epoxide, it hasn't been clear what that did. Did it just degrade the antioxidant properties, or did it even turn C60 into a prooxidant? Apparently it does neither.

 

Effects of Pin-up Oxygen on [60]Fullerene for Enhanced Antioxidant Activity

 

The introduction of pin-up oxygen on C60, such as in the oxidized fullerenes C60O and C60On, induced noticeable increase in the antioxidant activity as compared to pristine C60. The water-soluble inclusion complexes of fullerenes C60O and C60On reacted with linoleic acid peroxyl radical 1.7 and 2.4 times faster, respectively.

 

 

 

 

 

Edited by Turnbuckle, 15 August 2015 - 12:10 PM.

[Kalliste]

This could be an interesting article but I can't access it myself.

http://www.fasebj.or...275404.abstract

 

Mitochondrial-targeted antioxidants

1. Anne O. Oyewole and
2. Mark A. Birch-Machin¹

+ Author Affiliations
 

This Article

1. Published online before print August 7, 2015, doi: 10.1096/fj.15-275404 fj.15-275404

1. Dermatological Sciences, Institute of Cellular Medicine, Medical School, Newcastle University, Newcastle upon Tyne, United Kingdom

1. ↵1Correspondence: Dermatological Sciences, Institute of Cellular Medicine, Medical School, Framlington Place, Newcastle University, Newcastle upon Tyne, NE2 4HH, United Kingdom. E-mail: m.a.birch-machin@ncl.ac.uk

Abstract

Redox homeostasis is maintained by the antioxidant defense system, which is responsible for eliminating a wide range of oxidants, including reactive oxygen species (ROS), lipid peroxides, and metals. Mitochondrial (mt)-localized antioxidants are widely studied because the mitochondria; the major producer of intracellular ROS, has been linked to the cause of aging and other chronic diseases. Mt-targeted antioxidants have shown great potential because they cross the mt phospholipid bilayer and eliminate ROS at the heart of the source. Growing evidence has identified mt-targeted antioxidants, such as MitoQ and tiron as potentially effective antioxidant therapies against the damage caused by enhanced ROS generation. This literature review summarizes the current knowledge on mt-targeted antioxidants and their contribution to the body’s antioxidant defense system. In addition to addressing the concerns surrounding current antioxidant strategies, including difficulties in targeting antioxidant treatment to sites of pathologic oxidative damage, we discuss promising therapeutic agents and new strategic approaches.—Oyewole, A. O., Birch-Machin, M. A. Mitochondrial-targeted antioxidants.

 

 

 

Edited by Cosmicalstorm, 15 August 2015 - 12:29 PM.

[Kalliste]

Thymic involution is dependent on redox problems. This is nice news for those of us who are younger and still have the time to save it via bioavailable effective antioxidants. I was 30 when I started C60 so that was probably too late, if C60 can help

 

 

Rapid aging of the thymus linked to decline in free radical defenses Date: August 6, 2015 Source: Cell Press Summary: A critical immune organ called the thymus shrinks rapidly with age, putting older individuals at greater risk for life-threatening infections. A new study reveals that thymus atrophy may stem from a decline in its ability to protect against DNA damage from free radicals.

The damage accelerates metabolic dysfunction in the organ, progressively reducing its production of pathogen-fighting T cells.

 

A critical immune organ called the thymus shrinks rapidly with age, putting older individuals at greater risk for life-threatening infections. A study published August 6 in Cell

Reports reveals that thymus atrophy may stem from a decline in its ability to protect against DNA damage from free radicals. The damage accelerates metabolic dysfunction in the organ, progressively reducing its production of pathogen-fighting T cells.

The findings suggest that common dietary antioxidants may slow thymus atrophy and could represent a promising treatment strategy for protecting older adults from infections.

 

"The thymus ages more rapidly than any other tissue in the body, diminishing the ability of older individuals to respond to new immunologic challenges, including evolving pathogens and the vaccines that may otherwise offer protection from them," says senior study author Howard Petrie of the Scripps Research Institute. "We provide, for the first time, a mechanistic link between antioxidants and normal immune function, opening new avenues for potential treatment strategies that could improve immune defenses in the aging population."

 

The thymus produces essential immune cells called T cells, which are continuously lost and must be replaced throughout life. But starting around the time of puberty, the thymus rapidly decreases in size and loses its capacity to produce enough new T cells. This loss is partially offset by the duplication of existing T cells, but the resulting population of cells becomes more and more biased toward memory T cells, which recognize pathogens from previous or ongoing infections. As a result, broad-spectrum immunity against new pathogens and protective immune responses elicited by new vaccines diminish with age.

The development of interventions to slow the progression of thymus atrophy has been limited by the lack of knowledge about the underlying mechanisms. The prevailing theory suggests that sex hormones play a key role, but this explanation does not account for the accelerated speed at which the thymus diminishes in size in comparison to other tissues. Moreover, the body of scientific evidence clearly indicates that other factors must be involved in age-related thymus atrophy.

To address this question, Petrie and first author Ann Griffith, currently at the University of Texas Health Science Center at San Antonio, developed a computational approach for analyzing the activity of genes in two major thymic cell types--stromal cells and lymphoid cells--in mouse tissues, which are very similar to human thymic tissues in terms of function and the properties of atrophy. They found that stromal cells were deficient in an antioxidant enzyme called catalase, resulting in the accumulation of free radical and metabolic damage.

To test whether catalase deficiency plays a causal role in thymus atrophy, the researchers performed genetic experiments to enhance catalase levels in mice. By 6 months of age, the size of the thymus of the genetically engineered mice was more than double that of normal mice. Moreover, mice that were treated with two common antioxidants from the time of weaning achieved nearly normal thymus size by 10 weeks of age.

Taken together, the findings provide support for the free-radical theory of aging, which proposes that reactive oxygen species such as hydrogen peroxide cause cellular damage that contributes to aging and a variety of age-related diseases. These toxic molecules, which form in cells as a natural byproduct of the metabolism of oxygen, have been linked to progressive atrophy in many organs and tissues as part of the normal aging process. However, these are generally slow, progressive processes that do not become apparent until late in life and often go mostly unnoticed.

"In the case of the thymus, atrophy is more rapid than other tissues, which we now show is a consequence of stromal catalase deficiency in the context of a highly metabolic environment designed to support the demands of T-cell proliferation," Petrie says. "Our studies show that, rather than an idiosyncratic relationship to sex steroids, thymic atrophy represents the widely recognized process of accumulated cellular damage resulting from lifelong exposure to the oxidative byproducts of aerobic metabolism."

In future studies, the researchers will investigate whether antioxidant supplementation improves the functioning of the thymus and the immune system during aging. If these studies provide support for this idea, then they could lead to the development of new clinical recommendations for the prevention or treatment of age-related thymus atrophy in humans.

 

Griffith et al. Metabolic damage and premature thymus aging caused by stromal catalase deficiency. Cell Reports, August 2015 DOI: 10.1016/j.celrep.2015.07.008

 

The study in my quotation above showed that the Thymus could be protected by efficient antioxidants. That is interesting to ponder while reading the below quote:

 

Although there were statistically significant differenc-es in the following findings at the end of the administra-tion period, they were not considered to be toxicologicalbecause there was no dose-dependency; an increase in thenumber of positive incidences of urine ketone bodies inmales, a decrease in the differential lymphocyte ratio andan increase in the differential eosinophil ratio in malesin hematology, an increase in serum creatinine in males,and an increase in relative thymus weight in females.As for decreases in serum albumin and the relative kid-ney weight at 1,000 mg/kg/day in males at the end of theadministration period, and also an increase in total proteinat 1,000 mg/kg/day in females at the end of the recoveryperiod, their toxicological significance remains to be elu-cidated in the present study

 

 

Sub-acute oral toxicity study with fullerene C60 in rats

http://owndoc.com/pd...s non-toxic.pdf