I asked Ray once. This was his response.
Q: What is your opinion on Centrophenoxine/Meclofenoxate, piracetam or DMAE, for lipofuscin removal? The former particularly seems to have interesting effects on neurotransmitters in the brain.
A: I don't think there's nearly enough knowledge about its interactions with diet, stress, and hormones.
Exp Gerontol. 1991;26(4):365-74.
Effect of vitamin E on the accumulation of fluorescent material in cultured
cerebral cortical cells of mice.
Kan S, Devi SA, Kawashima S.
Zoological Institute, Faculty of Science, University of Tokyo, Japan.
The effect of vitamin E on the accumulation of lipofuscin-containing fluorescent
material in the mouse cerebral cortical cells in primary culture was studied.
Fluorescent material was extracted in ethanol:diethylether (3:1) and
autofluorescence intensity of the extracts was measured by a
spectrofluorophotometer. Although vitamin E at the concentration of 0.005 IU/ml
was not effective, 0.01 IU/ml vitamin E inhibited the accumulation of fluorescent
material. Fluorescent material accumulation was reduced to 76.3-86.4% of the
control level in 6-, 12-, or 18-day treatment of 0.01 IU/ml vitamin E. High doses
of vitamin E (0.05 or 0.1 IU/ml) were toxic for cultured cells. Ethanol, the
vehicle of vitamin E, at the final concentration of 0.005% was also effective on
the reduction of fluorescent material accumulation (81.0% of the control level at
18 days). The inhibitory effects of vitamin E as well as ethanol on the
accumulation of fluorescent material in cultured cells are explained by their
nature as free radical scavengers.
Exp Gerontol. 2002 Oct-Nov;37(10-11):1223-8.
Mild stress-induced stimulation of heat-shock protein synthesis and improved
functional ability of human fibroblasts undergoing aging in vitro.
Fonager J, Beedholm R, Clark BF, Rattan SI.
Laboratory of Cellular Ageing, Department of Molecular Biology, Danish Centre for
Molecular Gerontology, University of Aarhus, Gustav Wieds Vej 10-C, DK-8000
Repeated mild heat-shock (RMHS) treatment has anti-aging hormetic effects on
human fibroblasts undergoing aging in vitro. Since heat and various other
stresses induce the transcription and translation of heat-shock proteins (Hsp),
it was investigated if RMHS treatment affected the basal levels of four major
stress proteins Hsp27, 70, 90 and Hsc70. The basal levels of Hsp27, Hsc70, and
Hsp70 increased significantly in late passage senescent cells, which is
indicative of an adaptive response to cumulative intracellular stress during
aging. RMHS increased the levels of these Hsp even in early passage young cells
and were maintained high throughout their replicative lifespan. In comparison,
the amount of Hsp90 decreased both with aging and RMHS treatment in vitro.
However, whereas the difference in the levels of Hsp70 and Hsp90 was
statistically significant, the levels of Hsp27 and Hsc70 were statistically
similar in normal and RMHS-treated serially passaged cells. These alterations
were accompanied by an improved functional and survival ability of the cells in
terms of increased proteasomal activities, increased ability to decompose
H(2)O(2), reduced accumulation of lipofuscin and enhanced resistance to ethanol,
H(2)O(2) and UV-A radiation.
Free Radic Biol Med. 1989;7(6):611-6.
Effect of ethanol on lipofuscin accumulation in cultured rat cardiac myocytes.
Sohal RS, Marzabadi MR, Brunk UT.
Department of Biological Sciences, Southern Methodist University, Dallas TX
The objective of this study was to determine the effect of ethanol on in vitro
life span, rate of contraction and lipofuscin content of neonatal rat cardiac
myocytes. Lipofuscin was quantified by microspectrofluorometry. The effects of 0,
3.1, 6.5, and 12.5 mM ethanol on myocytes, kept under an ambient oxygen
concentration of 20% and 40%, were studied. Exposure to low concentrations of
ethanol resulted in a decrease in the amount of lipofuscin whereas exposure to
high concentration of ethanol caused an increase in the level of lipofuscin. The
length of cell survival in controls and 3.1 mM ethanol exposed myocytes was
similar under 20% oxygen, but was longer in the latter group under 40% oxygen, as
compared to controls. The total number of contractions in 3.1 mM ethanol-exposed
myocytes were, respectively, 4% and 8% higher under 20% and 40% oxygen atmosphere
than in control cells.
Pharmacology. 1978;16 Suppl 1:36-44.
Dihydroergotoxine(Hydergine) and ethanol-induced aging of C57BL/6J male mice.
Samorajski T, Rolsten C, Pratte KA.
The residual effects of dihydroergotoxine (Hydergine), ethanol, and
Hydergine-plus-ethanol were investigated in old male mice of the C57BL/6J strain.
Prolonged 10% v/v ethanol, concomitant with adequate nutrition, produced a
significant decrease in life-span, decreased spontaneous locomotor activity and
reduced lipofuscin accumulation in neurons of the reticularis gigantocellularis.
These effects were not apparent in mice treated with Hydergine or alcohol
combined with Hydergine.
Indian J Exp Biol. 1996 Aug;34(8):776-81.
Age-related decline in multiple unit action potentials of cerebral cortex
correlates with the number of lipofuscin-containing neurons.
Sharma D, Singh R.
Neurobiology Laboratory, School of Life Sciences, Jawaharlal Nehru University,
New Delhi, India.
The present study examined whether there is any obvious correlation between the
density of lipofuscin-containing neurons and the spontaneous neuronal action
potentials (Multiple Unit Activity, MUA) in the parietal cortex of the aging rat
brain. The results showed that MUA counts were decreased with age while the
number of lipofuscin-containing neurons was increased. The cortex with the
highest percentage of lipofuscin-containing neurons had the lowest MUA counts
while the cortex with the lowest percentage of lipofuscin-containing neurons had
the highest MUA counts. The inverse correlation between MUA and
lipofuscin-containing neuron number was also evident when the population of the
lipofuscin-containing neurons was pharmacologically altered in vivo by the
administration of anti-lipofuscin drug centrophenoxine. The inverse relationship
between MUA and the lipofuscin-containing neuron numbers is consistent with: (i)
the correlations of MUA with age-related changes in lipid peroxidation and
biochemically measured lipofuscin concentration, and (ii) the oxidative
stress-induced impairments of neuronal electrophysiology.
Eye (Lond). 2011 Apr;25(4):519-27. doi: 10.1038/eye.2011.7. Epub 2011 Feb 11.
Calcium overload is associated with lipofuscin formation in human retinal pigment
epithelial cells fed with photoreceptor outer segments.
Zhang L, Hui YN, Wang YS, Ma JX, Wang JB, Ma LN.
Department of Ophthalmology, Eye Institute of Chinese PLA, Xijing Hospital, The
Fourth Military University, Xian, Shaanxi, China.
PURPOSE: To investigate the role of Ca²(+) in lipofuscin formation in human
retinal pigment epithelial (RPE) cells that phagocytize bovine photoreceptor
outer segments (POSs).
METHODS: Cultured human RPE cells fed with 2 × 10⁷per l bovine POS were treated
with flunarizine, an antagonist of Ca²(+) channel, or/and centrophenoxine, a
lipofuscin scavenger. The Ca²(+) changes and lipofuscin formation were measured
with fluoresence dye Fluo-3/AM ester, laser scanning confocal microscopy (LSCM)
and flow cytometry (FCM). The activity of RPE cells was measured by methyl
thiazolyl tetrazolium (MTT) assay and argyrophilic nucleolar organizer regions
RESULTS: The Ca²(+) fluorescence intensity (CFI) of RPE cells fed with POS was
significantly increased compared with the controls (165.36 ± 29.92 U). It reached
a peak with 777.33 ± 63.86 U (P<0.01) at 12 h, and then decreased but still
maintained a high level of 316.90 ± 36.07 U (P<0.01) for 4 days. Flunarizine and
centrophenoxine significantly decreased the Ca²(+) overload to 227.18 ± 14.00 U
at 12 h and 211.06 ± 20.45 U at 4 days. FCM confirmed these changes. The drugs
also showed an inhibitory effect on the lipofuscin formation. The proliferation
rate of the cells fed with POS increased significantly. Both drugs had inhibitory
effects on the activity of the cultured cells. This tendency was confirmed by
CONCLUSIONS: The Ca²(+) inflow initiated lipofuscin accumulation in RPE cells fed
with POS. Flunarizine and centrophenoxine can decrease Ca²(+) overload and
lipofuscin formation in RPE cells, accompanied by maintaining cellular vitality.
Indian J Exp Biol. 1992 Jun;30(6):470-3.
'Dark' cell formation under protein malnutrition: process of conversion and
concept of 'semi-dark' type Purkinje cells.
James TJ, Sharma SP, Gupta SK, Patro IK.
Department of Zoology, Kurukshetra University, India.
This paper deals with some deleterious effects of protein malnourishment in rat
cerebellum. Severe protein deprivation enhanced the formation of 'dark' cells in
white rats. It is postulated that abnormal changes in the neuronal contents
induced by nutritional stress play a vital role in the formation of the 'dark'
cells through an intermediary stage, 'semi-dark' cells. Centrophenoxine a
lipofuscinolytic agent, however, seems to interfere with the process of formation
of 'dark' cells and/or helps reconversion of the 'dark' cells into the normal or
'light' type Purkinje cells.
J Gerontol. 1983 Sep;38(5):525-31.
Lipofuscin response to the "aging-reversal" drug centrophenoxine in rat retinal pigment epithelium and frontal cortex.
Katz ML, Robison WG Jr.
The effects of centrophenoxine on the lipofuscin contents of the retinal pigment epithelium (RPE) and frontal cortex of the brain were examined in senescent female Fischer rats. Rats (106 weeks old) were injected daily for 11 weeks with centrophenoxine (80 to 120 mg/kg body weight) or saline, and then sacrificed along with untreated 28- and 46-week-old controls. The number of lipofuscin granules seen in the RPE by light microscopy increased by 70% between 28 and 117 weeks of age in control animals. There was a concomitant age-related increase in lipofuscin specific fluorescence in the RPE. Centrophenoxine treatment neither reduced the amount of lipofuscin, nor altered the ultrastructural appearance of lipofuscin granules in the RPE. Between 28 and 117 weeks of age, there was an almost nine-fold increase in the lipofuscin content of the frontal cortex of control animals; centrophenoxine treatment failed to reverse this pigment accumulation.
The roles of vitamin E and unsaturated fatty acids in the visual process.
Robison WG, Kuwabara T, Bieri JG.
Laboratory of Vision Research, National Eye Institute, National Institutes of
Health, Bethesda, Maryland, MD 20205.
Relatively high proportions of long-chain, polyunsaturated fatty acids seem to be
required in rod photoreceptor membranes in order to provide the precise
microenvironment for the proper function of the visual pigment rhodopsin. At the
same time, such high levels of lipid unsaturation put the photoreceptor membranes
at a high risk for autoxidation. The antioxidant vitamin E which can minimize
autoxidation of polyunsaturated fatty acids is found in rather high
concentrations in the outer segment membranes. Dietary deficiency in vitamin E
induces disintegration of rod outer segment membranes, probably by increasing
autoxidation. Also, it greatly accelerates the accumulation of aging pigments in
the retinal pigment epithelium, probably because these lipofuscin granules do
indeed represent the end products of lipid peroxidation. Vitamin E supplements,
up to threefold normal levels, appear to provide no significant protection of the
retina from light damage produced either by short but acute or by long-term, low
level exposures to light. This is not consistent with current theories which
implicate lipid peroxidation in the destruction of rod outer segments in light
damaged retinas; more work is needed before any relation between retinal light
damage and vitamin E levels can be assessed. Surprisingly, the amount of
lipofuscin granule accumulation in the retinal pigment epithelium is influenced
dramatically by dietary levels of vitamin A. Even retinas lacking a source of
polyunsaturated fatty acids from rod outer segments still may accumulate massive
lipofuscin if dietary vitamin A is provided. Perhaps vitamin A, which has such a
dynamic relationship with the retinal pigment epithelium, becomes oxidized, and
then contributes to the formation of a lipofuscin-like pigment. Centrophenoxine,
a drug claimed to be effective in reversing the accumulation of age-related
lipofuscin in the central nervous system, has no obvious effect in the eye or
uterus in removing the lipofuscin granules induced by vitamin E deficiency.
Microperoxisomes are abundant in the retinal pigment epithelium, and may be
associated with rapid lipid turnover and/or utilization of lipid soluble
vitamins. Their potential roles, however, need further documentation and
clarification. Recently developed techniques and new discoveries in lipid
research open the way for many fruitful studies on the interactions and precise
roles of lipids and lipid-soluble vitamins in vision.
J Gerontol (1977) 32 (1): 38-45.
Effects of Dimethylaminoethanol Upon Life-span and Behavior of Aged Japanese Quail1
Arthur Cherkin, Michael J. Eckardt
Psychobiology Research Laboratory, Veterans Administration Hospital, Sepulveda, CA 91343 and Depts. of Anesthesiology & Psychiatry. UCLA. Psychobiology Research Laboratory, Veterans Administration Hospital Sepulveda, CA 91343.
The lysosome hypothesis of aging predicts that membrane stabilizers will extend life-span. Stabilizers containing the dimethylaminoethanol moiety (DMAE) have been reported to extend the life-span of drosophila and mice. We tested the prediction in Japanese quail (N = 15) by administering DMAE bitartrate (18.4 mg/kg/day) in the drinking water for 69 weeks, starting at 195 weeks of age. A matched control group (N = 14) received tartaric acid (4.0 mg/kg/day) in the water. Contrary to the prediction, the DMAE-treated group had a shorter life-span after start of treatment (49 weeks) than the controls (69 weeks). No significant differences between the groups were observed in body weight or daily fluid intake. Three behavioral studies were carried out on survivors at 243-249 weeks of age, namely; activity response to light-flash; sexual mounting response to a female quail; and classical conditioning of the heart rate. Aged quail differed from young-adults in changes in motor activity in response to light flashes. Aged quail appeared less responsive initially to reinforced conditioning trials and demonstrated extinction when light flash was not followed by electric shock. There were no detectable differences in latency to mount or in basal heart rate, either as a function of age or as a function of DMAE treatment.