Mois : septembre 2024

Researchers have tested several combinations of ingredients with anti-aging properties. Those combinations, including a 12-ingredient formulation created by NOVOS, helped to reduce DNA damage and oxidative stress in human skin cells in cultures [1].

Fighting multiple Hallmarks of Aging at once

Aging impacts multiple cellular processes, and these impacts can be assessed through biomarkers. The researchers of this paper used human skin cells (keratinocytes) as their targets for assessing the impact of the NOVOS supplement and six other formulations on DNA damage and oxidative stress markers. They believe that since skin is frequently exposed to DNA-damaging agents, such as UV light, it is the right choice for assessing these markers.

The tested supplement formulation created by NOVOS, a New York-based company, is a mix of 12 ingredients: pterostilbene, a resveratrol-related polyphenol derived from blueberries; glucosamine sulfate, a supplement used for osteoarthritis treatment; fisetin, a senotherapeutic; glycine, an amino acid; lithium aspartate, a mineral; calcium alpha-ketoglutarate, a small molecule present in the human body; magnesium malate, a natural substance found in apples; vitamin C, also known as ascorbic acid; L-theanine, an amino acid found primarily in green and black tea; hyaluronic acid; Rhodiola rosea root extract; ginger root extract; and a boost consisting of β-nicotinamide mononucleotide.

These nature-based ingredients, all of which have good safety profiles, were selected based on their anti-aging activity, documented role in counteracting multiple aging processes, or association with aging-related mechanisms. We have previously written about them and their selection process in more detail.

The authors explained that they tested combinations of different compounds because this approach could address multiple aging processes at once. A combination of compounds could also effectively reduce unwanted side effects that can appear when a single compound is used in high concentrations.

Reduction in DNA damage

Even beneficial ingredients can be toxic in excessive concentrations. Therefore, the researchers started their study by assessing the compounds’ highest safe concentrations to use. They followed up by testing the impact of supplement formulation on protecting cells from DNA damage, using tumor p53 binding protein 1 (53BP1) as a DNA damage marker.

The researchers treated 2D cell cultures with the six formulations and NOVOS. Following this treatment, they induced DNA damage with the genotoxic agent neocarzinostatin (NCS).

Data analysis revealed that treating cells with the six different formulations caused mild adverse effects on cells, as evidenced by higher numbers of DNA damage-related foci in some cells. NOVOS treatment did not cause that.

Cells pre-treated with NOVOS were less likely to have many of these foci after NCS administration, which suggests that NOVOS helps alleviate the DNA damage caused by NCS. However, it didn’t bring the damage down to the level of control cells that were not exposed to NCS.

The six different formulations also positively affected cells exposed to NCS. Those formulas “significantly prevented DNA damage, observed as reduced foci numbers, even with a stronger effect compared to the NOVOS’ original formulation itself.”

Based on this statistical analysis and the number of ingredients in each formulation, the researchers chose NOVOS, F1, F2, and F4 for the following experiments.

The first was a test of one more DNA damage marker, γ-H2AX, a marker that reflects double-strand breaks in the DNA. However, they performed it in the 3D skin cell cultures since 3D cultures better reflect the skin microenvironment and ”allow the formation of a cellular structure, like a tissue.” The results of this experiment suggested that all four of these formulations significantly reduced DNA damage compared to cells that were only exposed to NCS.

Notably, the concentration of NOVOS used in this experiment is lower than that of other formulations. Still, it can achieve similar effects in counteracting DNA damage as F1, F2, and F4. The authors suggest that this may be because of some synergistic effects between the NOVOS compounds.

Decreased oxidative stress

In the following test, the researchers aimed to assess the formulations’ ability to prevent oxidative damage to the cells. Intracellular reactive oxygen species (ROS) are difficult to measure. However, since many of them are converted to hydrogen peroxide, hydrogen peroxide can be used as a proxy for intracellular ROS level changes and oxidative stress.

The researchers performed this experiment using 2D and 3D cultures. Those cells were treated with different F1, F2, F4, and NOVOS formulation concentrations. Then, the researchers used the pro-oxidant medication to induce oxidative damage.

F1 and F4 formulations were able to significantly reduce intracellular hydrogen peroxide production following menadione treatment in 2D and 3D cell cultures. NOVOS was able to achieve this in 2D but not 3D culture.

Limitations

This study suggests that NOVOS, along with the F1, F2, and F4 combinations, are able to counteract some aging-related processes in skin cells.

While the researchers discussed the benefits of using a combination of compounds over a single compound, this research lacks an experiment that compares the combinations to single compounds. Such experiments would be very informative in testing whether the observed effects result from the combined effect of the multiple compounds or whether the observed effects come mainly from the action of a single compound and there is a negligible contribution from the remaining compounds, which would allow for simpler supplements. This is notable as the F1, F2, and F4 combinations with half the NOVOS compounds show similar effects as NOVOS in the experiments.

The authors also note that further investigation into specific molecular pathways involved in the observed processes is necessary.

One of the study authors is an employee of NOVOS Labs. However, in the Conflict of Interest, the authors note: ”The funder NOVOS Labs was not involved in the study design, collection, analysis, interpretation of data, of this article or the decision to submit it for publication.”

Disclosure: A portion of the profits and equity from NOVOS are being donated to nonprofits working in the longevity science space, which includes us here at Lifespan.io.

Our Vice President, Dr. Oliver Medvedik, is also a scientific consultant, putting him in good company with Dr. Joao Pedro Magalhaes, Dr. Pamela Maher, Dr. Avi Rosenbaum, and Dr. Matt Kaeberlein, names with whom regular readers may be familiar.

Literature

[1] Punzo, A., Perillo, M., Silla, A., Malaguti, M., Hrelia, S., Diogo Barardo, Cristiana Caliceti, & Lorenzini, A. (2024). Promising Effects of Novel Supplement Formulas in Preventing Skin Aging in 3D Human Keratinocytes. Nutrients, 16(16), 2770–2770.

A new observational study suggests that lower LDL cholesterol levels are not necessarily better. Instead, the ratio of triglycerides to HDL may be more important [1].

Putting common wisdom to the test

LDL cholesterol is largely considered harmful, but lower levels might not be beneficial according to a new study by researchers from the University of Pittsburgh and the University of South Florida.

The researchers based their analysis on electronic medical records that were derived from the University of Pittsburgh Medical Center’s healthcare system and dated 2000 to 2022. This allowed them to assemble a vast cohort of 178,000 patients aged 50 to 89.

The participants had to meet several criteria, including a lack of diabetes or statin therapy at baseline and during the first year of follow-up. Patients who died during the first year of follow-up or had outlier levels of total or LDL cholesterol were also excluded to mitigate reverse causation. The researchers divided the sample into six LDL-C categories: 30-79, 80-99, 100-129, 130-159, 160-189, and over 190 mg/dL.

The golden mean

Over a 6-year average follow-up time, intriguing results emerged. A U-shaped relationship was observed between LDL-C levels and 10-year overall mortality. The highest all-cause mortality (19.8%) was found in the 30-79 group. In the other groups, it was 14.7%, 11.7%, 10.7%, 10.1%, and 14.0%, respectively.

Compared to the referent group of 80-99 mg/dL, this translated into a 23% increase in mortality risk for the lowest LDL-C group. In the following three groups, within the 100-189 mg/dL range, the mortality risk was lower than in the referent group by 13%, 12%, and 9%, respectively. In the highest LDL-C group of over 190 mg/dL, the mortality risk was again higher than in the referent group – by 19%.

The results were broadly similar for atherosclerotic cardiovascular disease (ASCVD) risk. However, in this case, the highest risk was recorded for the group with the highest LDL-C levels (over 190 mg/dL). Another interesting finding was that in men, even in this subgroup, mortality risk was not significantly higher than in the referent subgroup.

The parameter that could

However, the researchers did find a parameter with a linear relationship with mortality risk: the ratio of triglycerides and HDL cholesterol (HDL-C), which is often considered beneficial. Total cholesterol to HDL cholesterol ratio (T-C/HDL-C) also seemed to be a better predictor of mortality than LDL-C.

“Compared with patients in the highest quintile of triglycerides/HDL-C ratio (value of ≥3.44), those in the lowest quintile (value of ≤1.06) had an estimated 24% lower risk of mortality,” the paper says. “Thus, in aggregate and irrespective of age, the secondary lipid measures of T-C/HDL-C ratio and triglycerides/HDL-C ratio appeared to be more predictive of mortality than LDL-C, and a triglycerides/HDL-C ratio of about 1 or lower appears to be optimal.”

The researchers note that the insurance industry seems to have already discovered this relationship [2]: both T-C and HDL-C are more widely used in life insurance policy underwriting than LDL-C. This is not the first time that insurance companies, financially incentivized to discover factors related to health and mortality risks, have provided clues for scientists.

As with any observational study, this one cannot definitely prove causality. However, it aligns with some, though not all, of the previous research on this subject [3]. The authors call for more studies, including those on statin use, that would take into account both statins’ effectiveness in preventing cardiovascular disease and their side effects, such as muscle pain and increased risk of type 2 diabetes [4].

In this analysis among primary prevention-type patients without diabetes aged 50–89 years not on statin therapy at baseline or within 1 year, we found no evidence of a gradient relationship between LDL-C and long-term mortality risk. Instead, we observed that within the entire LDL-C range of 100–189 mg/dL (about two-thirds of the total patient population), mortality risk was similar and slightly lower than the referent LDL-C category of 80–99 mg/dL. These data conflict with the prevailing belief that ‘lower LDL-C is better’ yet align with results from multiple studies.

Literature

[1] Kip, K. E., Diamond, D., Mulukutla, S., & Marroquin, O. C. (2024). Is LDL cholesterol associated with long-term mortality among primary prevention adults? A retrospective cohort study from a large healthcare system. BMJ open, 14(3), e077949.

[2] Fulks, M., Stout, R. L., & Dolan, V. F. (2009). Association of cholesterol, LDL, HDL, cholesterol/HDL and triglyceride with all-cause mortality in life insurance applicants. J Insur Med, 41(4), 244-253.

[3] Johannesen, C. D. L., Langsted, A., Mortensen, M. B., & Nordestgaard, B. G. (2020). Association between low density lipoprotein and all cause and cause specific mortality in Denmark: prospective cohort study. Bmj, 371.

[4] Mansi, I. A., Chansard, M., Lingvay, I., Zhang, S., Halm, E. A., & Alvarez, C. A. (2021). Association of statin therapy initiation with diabetes progression: a retrospective matched-cohort study. JAMA internal medicine, 181(12), 1562-1574.

Neurological researchers, in Aging Cell, have deepened our understanding of the brain, explaining how turning off a dopamine receptor may lead to better memory in older people.

Sometimes it’s better for the dopamine not to hit

With aging comes changes in how the brain processes dopamine [1]. This work focuses on specific dopamine receptors called D3Rs, which are less common than related D2Rs although much stronger in binding dopamine [2]. This potency has made them attractive targets in drug discovery, and prior work has focused on D3Rs for the treatment of such disorders as schizophrenia and depression [3].

One of the brain regions that contains D3Rs is the hippocampus [4], which is responsible for the formation of memories. Rodent studies have found that, unlike D2Rs, blocking D3Rs improves memory [5] and cognition [6], and the same appears to be true for people [7]. However, the reasons why blocking a receptor would lead to cognitive benefits are not clear, which is why these researchers took a closer look.

Cells and mice agree

This work began with an examination of CA3-CA1 synapses, which are very common targets in brain research, derived from the hippocampi of mice. Without a D3R blocker, giving light stimulation to these synapses creates a weak long-term connection, which takes time and reinforcement to become a more permanent one.

However, administering a D3R blocker makes even this light stimulation form a permanent connection much more quickly. Strong stimulation does normally cause these synapses to form a permanent connection, but even this was enhanced with a D3R blocker. These findings were confirmed with a strain of mice that has the genes for D3 reception knocked out. Further work found that these behavioral changes were post-synaptic: they did not affect how the neurons worked before the stimulation.

Turning to living animals, the researchers found that D3R blocking turned short-term memory into long-term memory much more readily. Normally, a light training regimen allows mice to distinguish between familiar and unfamiliar objects for six to eight hours [8], after which they forget about it. However, administering D3R blockers to the mice allowed them to distinguish between the objects 24 hours later, as if they had gone through a heavier training regimen. The same was true of D3R knockout mice: they gained long-term memories much more quickly.

Although memories are harder to form with age, D3R numbers were found to be decreased with age, but mostly in the pre-synaptic sites. Post-synaptic D3R sites were largely similar between adult and aged mice. Comparing the hippocampal neurons of D3R knockout mice and wild-type mice, the researchers found that the age-related loss of memory formation did not occur: these neurons’ ability did not seem to have diminished. These findings were confirmed in mice that had their D3R receptors blocked through genetics or chemical intervention. No sex differences were found.

These findings are impressive, and it may be that D3 receptor blockers might become an effective “memory pill” for people seeking to learn information quickly or people suffering from age-related memory problems. However, there may be significant neurological side effects to engraving more things in long-term memory that would normally be lost. Further work needs to be done to judge the effectiveness and safety of blocking D3 in animals and, ultimately, in people.

Literature

[1] Shohamy, D., & Wimmer, G. E. (2013). Dopamine and the cost of aging. Nature Neuroscience, 16(5), 519-521.

[2] Maramai, S., Gemma, S., Brogi, S., Campiani, G., Butini, S., Stark, H., & Brindisi, M. (2016). Dopamine D3 receptor antagonists as potential therapeutics for the treatment of neurological diseases. Frontiers in neuroscience, 10, 451.

[3] Kiss, B., Laszlovszky, I., Krámos, B., Visegrády, A., Bobok, A., Lévay, G., … & Román, V. (2021). Neuronal dopamine D3 receptors: translational implications for preclinical research and CNS disorders. Biomolecules, 11(1), 104.

[4] Li, Y., & Kuzhikandathil, E. V. (2012). Molecular characterization of individual D 3 dopamine receptor-expressing cells isolated from multiple brain regions of a novel mouse model. Brain Structure and Function, 217, 809-833.

[5] Watson, D. J., Loiseau, F., Ingallinesi, M., Millan, M. J., Marsden, C. A., & Fone, K. C. (2012). Selective blockade of dopamine D3 receptors enhances while D2 receptor antagonism impairs social novelty discrimination and novel object recognition in rats: a key role for the prefrontal cortex. Neuropsychopharmacology, 37(3), 770-786.

[6] Xing, B., Meng, X., Wei, S., & Li, S. (2010). Influence of dopamine D3 receptor knockout on age-related decline of spatial memory. Neuroscience letters, 481(3), 149-153.

[7] Gross, G., Wicke, K., & Drescher, K. U. (2013). Dopamine D 3 receptor antagonism—still a therapeutic option for the treatment of schizophrenia. Naunyn-Schmiedeberg’s archives of pharmacology, 386, 155-166.

[8] Blokland, A., & Sesia, T. (2023). Delay-dependent forgetting in object recognition and object location test is dependent on strain and test. Behavioural Brain Research, 437, 114161.