*******************************************************

http://groups.yahoo.com/group/aspartameNM/message/1329
aspartame or MSG affects circadian rhythms in rats, two studies,
P. Subramanian, T. Manivasagam et al 2004:
Murray 2006.04.27

Pharmaceutical Biology
(Formerly International Journal of Pharmacognosy)
Publisher: Taylor & Francis
Issue: Volume 42, Number 1 / February 2004 Pages: 1 - 7
URL: Linking Options DOI: 10.1080/13880200390500885

Circadian variations of biochemical variables in aspartame treated rats.
P. Rajasekar, P. Subramanian, T. Manivasagam
Department of Biochemistry, Faculty of Science Annamalai University
Annamalainagar Tamil Nadu, India

[ Perumal Subramanian, Thamilarasan Manivasagam ]

Abstract:
In a study of the impact of aspartame on biochemical oscillations,
we examined the rhythms of blood glucose, plasma cholesterol,
and protein and serum aspartate transaminase (AST) in experimental rats.
Our results showed acrophase delays in glucose, total protein
and advances in AST rhythms and increased mesor (in AST),
amplitude (in cholesterol) and decreased amplitude values
(in glucose, AST) in aspartame treated animals.
Oral administration of aspartame might lead to increased levels
of aspartate in brain which could alter the characteristics of biochemical
variables possibly by modulating the transmission in several areas/nuclei
in brain including retinohypothalamic tract (RHT)
and suprachiasmatic nuclei (SCN).

Keywords:
Aspartame, Biochemical Variables, Circadian Rhythms

http://journalsonline.tandf.co.uk/media/8cac231aqh5tqlaf8q9x/contributions/a/a/d\
/h/aadhh4gxye1glka2.pdf

Accepted: May 4, 2004
Address correspondence to: P. Subramanian,
Department of Biochemistry,
Annamalai University, Annamalai Nagar - 608 002, Tamil Nadu,
India. E-mail: psub@...
DOI: 10.1080/13880200390500885 © 2004 Taylor & Francis Ltd.

Abstract
In a study of the impact of aspartame on biochemical oscillations,
we examined the rhythms of blood glucose, plasma cholesterol,
and protein and serum aspartate transaminase (AST)
in experimental rats.
Our results showed acrophase delays in glucose, total protein
and advances in AST rhythms
and increased mesor (in AST), amplitude (in cholesterol) and
decreased amplitude values (in glucose, AST)
in aspartame treated animals.
Oral administration of aspartame might lead
to increased levels of aspartate in brain which could alter the
characteristics of biochemical variables possibly by modulating
the transmission in several areas/nuclei in brain including
retinohypothalamic tract (RHT) and suprachiasmatic nuclei (SCN).

Keywords: Aspartame, biochemical variables, circadian rhythms.

Introduction

The biological clock in mammals is located in the suprachiasmatic
nuclei (SCN). (Hannibal, 2002).
Circadian clocks govern the timing of development, behaviour,
physiology, endocrinology and biochemistry, as well as photoperiodic
events (Forster et al., 2001).
These biological rhythms are adjusted daily (entrained) to the
environmental light/dark cycles via retinohypothalamic tract (RHT)
(Hannibal, 2002).
Many putative neurotransmitters have been identified in the
SCN (Vandenpol, 1980).
Aspartate has been reported to be a
putative excitatory neurotransmitter in retina
(Yagub & Eldred, 1991) in the RHT (Honma et al., 1996)
and SCN (Liou et al., 1986; Csaki et al., 2000).
This excitatory amino acid is involved in the transmission of light
information from retina to SCN via RHT
(Takeuchi & Takahashi, 1994).
Further, derivatives of aspartate (like N-acetyl aspartyl glutamate)
also act as a neurotransmitter in RHT (Hannibal, 2002).
Ingestion of L-aspartate into the SCN results in minor
phase advances during a subjective day
(Devries & Meijer, 1991).

Aspartame (ASP) is a dipeptide artificial sweetener;
on oral administration, it is hydrolysed in the gastrointestinal
tract to its constituent amino acids,
L-phenylalanine and L-asartic acid (Ranney & Oppermann, 1979).
Previous studies showed that dietary aspartame could alter the
diurnal feeding patterns, meal size, distribution of diurnal pattern of
spontaneous motor activity (Torie et al., 1985).

ASP reduced aggressive attack via a serotonergic meachanism
(Goerss et al., 2000).
The influences of chronic ASP ingestion
on brain neuropeptide Y (NPY) concentration, plasma hormone,
food intake and body fat in rats have been reported
(Beck et al., 2002).
The circadian nature of cholesterol synthesis
(Jones & Schoeller, 1990) and total protein rhythms in humans
and mice (Berezkin et al., 1992)
and circadian variation of AST (Coll et al., 1993) were documented.

However, the influences of ASP on biochemical circadian rhythms
has not been investigated intensively.
In the present study, ASP administration to Wistar rats
and its influence on circadian rhythms of blood variables
glucose, cholesterol, total protein and AST) were monitored.

Materials and methods

Animals

Male Wistar rats (180-190 g) were obtained from the Central
Animal House, Annamalai University.
They were housed in polypropylene cages and provided food pellets
(Agrocorporation Private Limited, Bangalore, India)
as a basal diet during the experiment.
Animals were maintained at room tempertature (25 ± 2 °C)
and under LD (12 : 12) conditions
(Rajakrishnan et al., 1999; Subramanian & Balamurugan,
1999; Subramanian et al., 1998, 2000, 2001).
Food and water were available ad libitum
throughout the experimental period.
Aspartame was obtained from SRL and all other biochemicals
used in the study were of analytical grade.

Treatment schedule

The animals were randomized and grouped into experimental
and control rats (n = 6 in each group).
Group I rats act as control, which received standard food pellets.
Group 2 animals were treated orally with aspartame
(500 mg/kg body weight) every day
(Sharma & Coulombe, 1987; Goerss et al., 2000)
throughout the experimental period (11 weeks).

Biochemical oscillations

At the end of the experimental period, a minimal amount of
blood (0.75ml) was collected from the orbital sinus using
heparinized tubes from normal and aspartame treated groups
every 4 hour (00:00, 04:00, 08:00, 12:00, 16:00, 20:00 and
24:00 hour) throughout the 24 hour period
(Subrmanian & Balamurugan, 1998; Subramanian et al., 2000).
Blood glucose (Fings et al., 1970),
plasma cholesterol (Zlatkis et al., 1953),
protein (Lowry et al., 1951)
and serum AST (Reitman & Frankel, 1957)
were estimated after blood collection.
At the end of the study both control and ASP treated rats
were killed by decapitation
and aspartate levels in brain tissues (Pfleiderer, 1969) were measured.

Figure 1. Temporal oscillations of glucose at 4 hour intervals for a period
of one day in Wistar rats [control (1A) and aspartame treated (1B)].
Dotted lines represent the raw data and solid lines represent the
best fitting cosinor curves
(obtained using "cosinorwin" computer software program).
Note 6 hour delay of the acrophase in aspartame treated animals
(Figure 1B).

Time series analysis

Time series analysis of the oscillation
(measurements of acrophase, amplitude and mesor)
were done by using "cosinorwin" computer software program.
Acrophase is the measure of peak time of the total rhythmic variability
in a 24 hour period.
mplitude corresponds to half of the total rhythmic variability in a cycle.
Mesor (M) is the rhythm adjusted mean.
It is equal to the arithmetic mean for equidistant data
covering the 24 hour period.
The acrophase is expressed in h;
mesor and amplitude values are expressed with the same
units as the documented variables.

Table 1. Changes in the rhythm characteristics of glucose
and cholesterol in control and aspartame treated rats.

Acrophase Amplitude Mesor
Biochemical variable Groups (h) (mg/dl) (mg/dl) r-value p-value

Glucose I 00:35 7.3 109.1 0.71dr (p < 0.05)
---------II 7.32 5.3 106.2 -0.34ns (p < 0.50)

Cholesterol I 18:41 8.2 51.9 -0.67dr (p < 0.05)
-----------II 18:49 8.7 51.3 -0.67dr (p < 0.05)

dr -- detectable rhythmicity.
ns -- no significant rhythmicity.

Figure 2. Diurnal rhythms of cholesterol at 4 hour intervals
for a period of one day in Wistar rats [control (2A)
and aspartame treated (2B)].
Note modulatory increase in amplitude of the rhythm.
No significant changes in acrophase was observed
in aspartame treated animals (Figure 2B).
Other details as in Figures 1A and 1B.

Results

Aspartate levels in brain tissues (mean ± SD) were increased
(4.212 ± 0.41) significantly (p < 0.001) in aspartame treated
animals when compared to controls (1.40 ± 0.15).

The endogenous circadian rhythms of glucose showed acrophase
at 00:35 hour in normal rats;
in the case of aspartame treated animals (group II),
maximum levels were found at 7:32 hour (Figure 1A, B).
The amplitude, mesor and r-values
were decreased significantly in group II
when compared with controls (group I).
Consinor analysis revealed detectable rhythmicity
in normal group and this rhythmicity is affected in
aspartame treated group (Table 1).

Cholesterol levels were maximum at 18:41 hour in normal
and at 18:49 hour in aspartame treated animals (Figure 2A, 2B).
Increased amplitude and decreased mesor values were found in
aspartame treated rats than that of normal ones.
Detectable cholesterol rhythms
were observed in two groups (Table 1).

Table 2. Changes in the rhythm characteristics of total protein and AST
in control and aspartame treated rats.

Acrophase Amplitude Mesor
Biochemical variable Groups (h) (mg/dl) (mg/dl) r-value p-value

Total protein I 08:48 1.0 5.0 0.02ns (p < 0.5)
-------------II 12:00 0.9 5.2 0.97dr (p < 0.001)

AST I 23:49 10.1 50.9 0.88dr (p < 0.002)
------------II 12:33 4.0 61.9 0.91dr (p < 0.001)

dr -- detectable rhythmicity.
ns -- no significant rhythmicity.

Figure 3. Temporal oscillations of total protein
at 4 hour intervals for a period of one day in Wistar rats
[control (3A) and aspartame treated (3B)].
Note 4 hour delay of the acrophase in aspartame treated animals
(Figure 3B).
Other details as in Figures 1A and 1B.

Total protein levels showed the peak value at 8:48 hour
and maximum value of aspartame treated animals was
observed at 12:00 hour (Figure 3A, 3B).
The amplitude and mesor values did not alter significantly in both cases.
The rvalue is increased in treated animals than that of normal
(Table 2).
Analysis of serum AST activity over the 24 hour period
revealed that the maximum activity
at 23:49 hour in normal and
at 12:33 hour in aspartame treated animals
(Figure 4A, B).
Low amplitude values and increased mesor
and r-values were observed in aspartame treated animals,
when compared with normal animals (Table 2).

Discussion

Neurotransmitters involving in regulation of circadian rhythms
were normally used to probe the nature of circadian rhythms.
Presence of aspartate in RHT (Liou et al., 1986),
horizontal, amacrine and ganglion cells, in some photoreceptors
and in some unidentified cells in the peripheral retina
(Yagub & Eldred, 1991) was reported.
The biochemical parameters chosen for this study showed
marked fluctuations over a 24 hour period.
From this study it can be concluded that
light dark cycles are the most effective synchronizers
for biochemical circadian rhythms studied in Wistar rats.

In the present study, peak time of glucose was at 3:00 hour
in normal and at 7:32 hour in treated animals;
this could be attributed to the food intake, digestion,
and accumulation of glucose in blood.
Rats administered aspartame with showed about 3 hour delays
in glucose rhythms.
However, no mesor value change was observed in group II
when compared with normal rats.
Aspartame could not alter glycemia (Ngugen et al., 1998).

Cholesterol levels in rats in the present study were increased
at night as reported previously (Ueberberg et al., 1984).
The whole body, free and total cholesterol syntheses
oscillate periodically and predictably (Jones & Scholler, 1990).

In rats, the rate limiting enzyme (HMG CoA reductase)
in the cholesterol synthesis pathway peak its activity at
midnight (Mietenen, 1982; Pappu & Illingworth, 1994).

The peak level of transcription of cholesterol-7a-hydroxylse
(7aH) gene was reported to occur in the evening.
All these factors may contribute to the nocturnal increase of
cholesterol.

Figure 4. Temporal variation of AST activity at 4 hour intervals for a
period of one day in Wistar rats
[control (4A) and aspartame treated (4B)].

Note 11 hour advance of the acrophase in aspartame treated animals
(Figure 4B).
Other details as in Figures 1A and 1B.

Circadian rhythms in total protein were reported in
humans and mice
(Touitou et al., 1986; Berezkin et al., 1992).
The positive and negative balance between synthesis
and degradation of protein might be responsible for the
rhythmic phenomenon.
In our experiment, AST levels are
maximum at 23 hour in control
and 11 hour advance in aspartame treated animals.
The significant increase in mesor and
amplitude of AST rhythm indicated that this may be due to
the aspartame metabolites;
aspartate might enter the TCA cycle via transamination of aspartate
to oxaloacetate (Ranney & Oppermann, 1979).

In the present study, administration of aspsartame
increased the brain aspartate levels corroborating
the previous results (Moller, 1991; Burns et al., 1991).
This increased brain aspartate might favour the transmission of
light information to the SCN (Takeuchi & Takahashi, 1994)

Further, aspartame is known to act on NMDA receptors
and has some neurological effects seen with glutamate
(Disk, 2000; Abdollahi et al., 2001).

Activation of NMDA receptors tends to transmit photic information
to the SCN (Mintz et al., 1999).
This activation leads to increased production of nitric oxide (NO)
(Meller & Gebhart, 1993).
Involvement of NO in the transmission of light information to SCN
is also suggested (Caillol et al., 2000).

Previous results showed that administration of aspartate into the SCN
induced phase shifts in the free running rhythms of hamsters
(Smith, 2000).

Hence, we speculate that increased aspartate levels in brain
could alter the characteristics of biochemical rhythms
studied, possibly by modulating the transmission in several
areas/nuclei in brain including RHT and SCN.

References

Abdollahi M, Nikfar S, Abdoli N (2001):
Potentiation by nitric oxide synthase inhibitor
and calcium channel blocker of aspartame
induced antinociception in the neouse formalin test.
Funda Clin Pharmacol 15: 117-123. mabdol@...

Beck B, Burlet A, Max JP, Stricker-Krongrad A. (2002):
Effects of long-term ingestion of aspartame on hypothalamic
neuropeptide Y, pasma leptin and body weight gain and composition.
J Physiol Behav 75: 41-47. bernard.beck@...;

Berezkin MV, Grastisinskii EN, Kudinova VF, Batygov AN,
Ponomareve LE, Prikazchikova O, Zhunkova GN (1992):
Seasonal and circadian fluctuactions in blood biochemical
indicators in mice in natural conditions and exposed to constant light.
Bull EKSP Biol Med 114: 75-78.

Burns TS, Stargel WW, Tsehanz C, Kotdonis FN, Hurwitz A (1991):
Aspartame and sucrose produce a similar increase
in the plasma phenylalanine to large neutal amino acid ratio
in healthy subjects. Pharmacology 43: 210-219.

Caillol M, Devinoy E, Lacroise MC, Schirar A (2000):
Endothelial and neuronal nitric oxide synthases are present in the
suprachiasmatic nuclei of syrian hamsters and rats.
Eur J Neurosci 12: 649-61.

Coll AR, Arderiux F, Noguera AD (1993):
Circadian rhythms of serum concentrations of 12 enzymes
of clinical interest. Chronobiol Int 10: 190-202.

Csaki A, Kocsis K, Halasz B, Kiss J (2000):
Localization of glutamatergic/aspartatergic neurons projecting
to the hypothalamic paraventricular nucleus
studied by retero grade transport. J Neurosci 101: 637-655.

Devries MJ, Meijer JH (1991):
Aspartate injections into the suprachaismatic region
of Syrian hamsters do not mimic
the effects of light on the circadian activity rhythys.
Neurosci Lett 127: 215-218.

Disk (2000):
Micromedex international health series.
Aspartame neurologic clinical effects. Toxicol Inf Poison: 105.

Fings CS, Toltiff CR, Duonin RT (1970):
Glucose determination by o-toluidine method
using glacial acetic acid.
In: G Toro PG Ackermann eds., Practical Clinical Chemistry.
Boston. Little Brown and Bronon CO 115-118.

Forster CH, Winter C, Hofbauer A, Hall JC,
Stanewsky R (2001):
The circadian clock of fruit flies is blind after
elimination of all known photoreceptor.
J Neuron 30: 249-261.

Goerss AL, Wagner GC, Wendy L (2000):
Acute effects of aspartame on aggression and neurochemistry of rats.
Life Sci 67: 1325-1329.

Hannibal J (2002):
Neurtransmitters of the retinohypothalamic tract.
J Cell Tissue Res 309: 73-88.

Honma S, Katsuno Y, hinohara K, Abe H, Honma K [1996]:
Circadian rhythm and response to light of extracellular glutamate
and aspartate in rat suprachiasmatic nucleus.
Am J Physiol 271: [Regulatory Integrative Comp Physiol 40]
R579-R585.

Jones PJH, Schoeller DA (1990):
Evidence of diurnal periodicity in human cholesterol synthesis.
J Lipid Res 37: 667-693.

Liou SY, Shibata S, Iwasaki K, Ueki (1986):
Optic nerve stimulation-induced increase of release of 3H-glutamate
and 3Haspartate but not 3H-GABA from the suprachiasmatic
nucleus in slices of rat hypothalamus.
Brain Res Bull 16: 527-531.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951):
Protein measurement with the Folin-phenol reagent.
J Biol Chem 193: 265-275.

Meller ST, Gebhart GF (1993):
Nitric oxide (NO) and nociceptive processing in the spinal cord.
Pain 2: 127- 136.

Mietencen TA (1982):
Diurnal variation of cholesterol precursors
squalene and methyl sterols in human plasma lipoproteins.
J Lipid Res 23: 466-473.

Mintz EM, Marvel CL, Gillespie CF, Price KM,
Albers HE (1999):
Activation of NMDA receptors in the suprachiasmatic
nucleus produces light-like phase shifts of the circadian
clock in vivo. J Neurosci 19: 5124-5130.

Moller SE (1991):
Effect of aspartame and protein administered
in phenylalanine equivalence doses, on plasma netural
amino acids, aspartate, insulin, and glucose in man.
Pharmacol Toxicol 68: 408-412. sem@...

Nguyen UN, Dumoulin G, Henriet MT, Regnard J (1998):
Aspartame ingestion increases urinary calcium, but not
oxalate excretion, in healthy subjects.
Clin Endocrinol Metabol 83: 165-168.

Pappu AS, Illingworth DR (1994):
Diurnal variations in the plasma concentrations of mevalonicacid
in patients with abeta lipoproteinaemia.
Eur J Clin Invest 24: 698-702.

Pfleiderer G (1969):
In: Methods of Enzymatic Analysis.
Bergmeyer HU Ed, New York and London,
Academic Press, pp. 381-383.

Rajakrishnan V, Subramanian P, Viswanathan P,
Menon VP (1999):
Effect of chronic ethanol ingestion on biochemical
circadian rhythms in Wistar rats.
Alcohol 18: 147-152.
[ Chronic ingestion of ethanol for 60 days was known to alter
the characteristics of biochemical circadian rhythms in Wistar rats.
Peak times of glucose, potassium and lactic acid rhythms
were delayed by 18 h, 3 h, and 3 h respectively,
whereas peak times of cholesterol and malondialdehyde rhythms
were advanced by 3 h and 9 h respectively during ethanol treatment.
Significant changes in range (p < 0.001 expect in calcium)
and 24 h mean (p < 0.001) of all the biochemical circadian rhythms
studied were observed during ethanol treatment.
The alterations in the characteristics of these biochemical
circadian rhythms could be principally due to the alterations
on the hepatic cellular architecture;
other plausible underlying reasons are also discussed.
PMID: 10456565 ]

Ranney RE, Oppermann JA (1979):
A review of the metabolism
of the aspartyl moiety of aspartame in experimental animals
and man. J Environ Pathol Toxicol 2: 979-985.

Reitman S, Frankel AS (1957):
A colorimetric method for the determination of serum glutamic,
oxaloacetic and glutamic pyruvic transaminases.
Am J Clin Path 28: 53-56.

Sharma KP, Coulombe RA (1987):
Effects of intense sweeteners on hunger, food intake and body weight.
Food Chem Toxicol 25: 565-568.

Smith RQ (2000):
Transport of glutamate and other aminoacids at the blood brain barrier.
J Nutr 30: 1016S-1022S.

Subramanian P, Balamurugan E (1999):
Temporal oscillations of serum electrolytes
in N-phathaloyl GABA-treated rats.
Pharmacol Biochem Behav 62: 511-514.
[ N-Phthaloyl gamma-aminobutyric acid (P-GABA) has been known
to cross the blood-brain barrier
and ultimately to increase brain GABA level.
In the present study, P-GABA was administered to Wistar rats
for 21 days and
circadian rhythms of sodium, potassium, and calcium levels in serum
were studied under seminatural light-dark conditions.
P-GABA administration caused desynchronization of the rhythms
and advanced the peak times of serum electrolytes.
Exogenously administered P-GABA could alter the photic information
received by the clock.
The results could be explained by slightly less or more than
1-h daily delays, which would bring the peak times to the points
21 days after the start of administration.
The changes in amount of electrolytes after P-GABA
administration are discussed. PMID: 10080244 ]

Subramanian P, Menon VP, Arokiam FV, Rajakrishnan V (1998):
Lithium modulates biochemical circadian rhythms in Wistar rats.
Chronobiol Int 15: 29-38.
[ We studied the effects of chronic lithium treatment
on circadian rhythms of
glucose, cholesterol, calcium, potassium, malondialdehyde (MDA),
and lactic acid in Wistar rats.
Lithium altered the peak time, range, and 24h mean
of these biochemical rhythms.
Peak times of the circadian rhythms of
glucose, calcium, and potassium were delayed by 3h, 6h, and 6h,
respectively, whereas circadian rhythms of MDA and lactic acid
were advanced by 9h and 3h, respectively, in lithium-treated rats.
Delays observed in our experiments would support the hypothesis
that lithium's therapeutic effect is to delay overtly fast circadian
rhythms.
Advances of peak times owing to lithium treatment are discussed.
PMID: 9493712 ]

Subramanian P, Sivabalan S, Menon VP, Vasudevan K (2000 March)
Influence of chronic zinc supplementation on biochemical
variables and circadian rhythms in Wistar rats.
J Nutr Res 20(3): 413-425.

Subramanian P, Sundaresen S, Balamurugan E (2001):
Temporal oscillations of phosphatases
in N-phthaloyl gammaaminobutyric acid treated rats.
Indian J Exptl Biol 36: 1141-1143.
[ N-pathaloyl gamma-aminobutyric acid (P-GABA)
was administered to Wistar and 24 hr rhythms
of acid and alkaline phosphatases were studied
under light-dark conditions.
P-GABA administration advanced the peak times of phosphatases.
Since GABA is being involved in conveying dark information
to the clock, exogenous administration of P-GABA
might reduce the photic information received by the clock.
The results could be explained by slight daily advances
which would bring the peak times to the points 21 days
after the start of administration. PMID: 10085783 ]

Takeuchi Y, Takahashi K (1994):
Circadian variations of amino acid content
of suprachiasmatic nucleus in rats.
Neuroscience Lett 178: 275-278.

Torie K, Mimura T, Takasaki Y, Ichimura M (1985):
Dietary aspartame with protein on plasma and brain amino acids,
brain monoamines and behaviour in rats.
J Physiol Behav 36: 765-771.

Touitou Y, Rein berg A, Bogdan A, Auzeby A, Beck H,
Touitou C (1986):
Differences between young and elderly subjects
in variations of total plasma proteins and blood volume as
reflected by hemoglobin, hematocrit and erythrocyte counts.
Clin Chem 32: 801-804.

Ueberberg H, Laque K, Trieb G (1984):
Comparative studies on
the circadian rhythm of conticosterone lipid and cholesterol
levels in adrenals and blood of rats.
Chronobiol Int 1: 41-49.

Vandenpol AN (1980):
The hypothalamic suprachiasmatic nucleus of the rat intrinsic anatomy.
J Comp Neurol 191: 661-702.

Yagub A, Eldred WD (1991):
Localization of aspartate like immuno reactivity
in the retina of the turtle (Pseudemys scripta).
J Comp Neurol 312: 584-598.

Yokogoshi H, Roberts CH, Caballero B, Wurtman RJ (1984):
Effects of aspartame and glucose administration on brain
and plasma levels of large neutral amino acids
and brain 5-hydroxy indoles. Am J Clin Nutr 40: 1-7.

Zlatkis A, Zak B, Boyle AJ (1953):
A new method for the direct determination of serum cholesterol.
J Lab Clin Med 45: 486.

[ Indian J Exp Biol. 2003 Aug; 41(8): 797-804.
Circadian clock genes in Drosophila: recent developments.
Subramanian P, Balamurugan E, Suthakar G. psub@...
Department of Biochemistry, Faculty of Science,
Annamalai University, Annamalainagar 608 002, India.

Circadian rhythms provide a temporal framework to living organisms
and are established in a majority of eukaryotes
and in a few prokaryotes.
The molecular mechanisms of circadian clock
is constantly being investigated in Drosophila melanogaster.
The core of the clock mechanism was described by a
transcription-translation feedback loop model involving period (per),
timeless (tim), dclock and cycle genes.
However, recent research has identified multiple feedback loops
controlling rhythm generation and expression.
Novel mutations of timeless throw more light
on the functions of per and tim products.
Analysis of pdf neuropeptide gene
(expressed in circadian pacemaker cells in Drosophila),
indicate that PDF acts as the principal circadian transmitter
and is involved in output pathways.
The product of cryptochrome is known to function as a circadian
photoreceptor as well as component of the circadian clock.
This review focuses on the recent progress in the field
of molecular rhythm research in the fruit fly.
The gene(s) and the gene product(s) that are involved
in the transmission of environmental information to the clock,
as well as the timing signals from the clock outward to cellular functions
are remain to be determined.
Publication Types: Review PMID: 15248475 ]

[ Pharmaceutical Biology (Formerly International Journal of Pharmacognosy)
Publisher: Taylor & Francis
Issue: Volume 43, Number 3 / April-May 2005 Pages: 209 - 218
URL: Linking Options DOI: 10.1080/13880200590928771

Influence of garlic extract on temporal characteristics of lipid
peroxidation
products and antioxidants in tumor-bearing rats.
P. Subramanian, S. Sundaresan, T. Manivasagam
Department of Biochemistry, Faculty of Science,
Annamalai University, Annamalainagar, Tamil Nadu, India

Abstract:
Effects of garlic (Allium sativum L.) extract on the temporal patterns
of circulatory lipid peroxidation products and antioxidants during
N-nitrosodiethylamine (NDEA)-induced hepatic tumorigenesis
were investigated in Wistar rats.
Experimental animals were divided into control, NDEA-treated,
NDEA + garlic-treated, and garlic-treated.
The characteristics of circadian rhythms
(acrophase, amplitude, and mesor) of lipid peroxidation products
(thiobarbituric acid reactive substances) and antioxidants
(reduced glutathione, glutathione peroxidase, superoxide dismutase,
and catalase) were analyzed.
Alterations in the temporal characteristics of all variables
were observed in NDEA-treated rats when compared to other groups.
The alterations in the characteristics of these variables
in NDEA-treated and in other groups deserve further investigation
for the prognosis and therapeutic efficacy of chronotherapy of cancer.

Keywords:
Allium sativum, antioxidants, circadian rhythms, hepatocarcinogenesis,
N-nitrosodiethylamine ]
*******************************************************


http://www.if-pan.krakow.pl/pjp/pdf/2004/1_79.pdf free full text
Polish J. Pharmacol., 2004, 56, 79-84. ISSN 1230-6002
Monosodium glutamate affects the temporal
characteristics of biochemical variables in Wistar rats.
Thamilarasan Manivasagam, Perumal Subramanian

Monosodium glutamate (MSG) was administrated chronically for 60
days to Wistar rats
and 24 h rhythms of glucose, Cholesterol, total protein and
alkaline phosphatase were studied.
MSG treatment was found to cause acrophase delays in the glucose
and alkaline phosphatase rhythms and advances in acrophases of
cholesterol and total protein levels.
Amplitude and mesor values of these rhythms were found to be
altered during MSG treatment.
Glutamate levels in the brain were found to be significantly increased,
which could alter these biochemical rhythms by modulating the
transmission in retinohypothalamic tract and in the hypothalamic nuclei,
probably including suprachiasmatic nuclei.

Key words: circadian rhythm, monosodium glutamate, glucose,
cholesterol, total protein, alkaline phosphatase

"MSG, 600 mg/kg (4) was injected subcutaneously to group II rats
every day (at irregular intervals) for 60 days." [ 6 rats ]
*******************************************************

http://groups.yahoo.com/group/aspartameNM/message/1328
migraine from sucralose, Bigal ME & Krymchantowski AV,
Headache 2006 March; formaldehyde from 11% methanol part of
aspartame or from red wine causes same toxicity (hangover) harm:
Murray 2006.04.24

"Of course, everyone chooses, as a natural priority,
to actively find, quickly share, and positively act upon the facts
about healthy and safe food, drink, and environment."

Rich Murray, MA Room For All rmforall@...
505-501-2298 1943 Otowi Road Santa Fe, New Mexico 87505

http://groups.yahoo.com/group/aspartameNM/messages
group with 151 members, 1,329 posts in a public, searchable archive
http://RMForAll.blogspot.com http://AspartameNM.blogspot.com

Dark wines and liquors, as well as aspartame, provide
similar levels of methanol, above 120 mg daily, for
long-term heavy users, 2 L daily, about 6 cans.

Within hours, methanol is inevitably largely turned into formaldehyde,
and thence largely into formic acid -- the major causes of the dreaded
symptoms of "next morning" hangover.

Fully 11% of aspartame is methanol -- 1,120 mg aspartame
in 2 L diet soda, almost six 12-oz cans, gives 123 mg
methanol (wood alcohol). If 30% of the methanol is turned
into formaldehyde, the amount of formaldehyde, 37 mg,
is 18.5 times the USA EPA limit for daily formaldehyde in
drinking water, 2.0 mg in 2 L average daily drinking water.

http://groups.yahoo.com/group/aspartameNM/message/1143
methanol (formaldehyde, formic acid) disposition: Bouchard M
et al, full plain text, 2001: substantial sources are
degradation of fruit pectins, liquors, aspartame, smoke:
Murray 2005.04.02

Any unsuspected source of methanol, which the body always quickly
and largely turns into formaldehyde and then formic acid, must be
monitored, especially for high responsibility occupations, often with
night shifts, such as pilots and nuclear reactor operators.

http://groups.yahoo.com/group/aspartameNM/message/1291
European Food Safety Authority to decide aspartame safety by May:
caffeine diet drinks cause female hypertension, WC Winkelmayer et al,
JAMA 2005.11.09: PubMed lists 50 items for "diet soft drinks" since
2004 Oct.: Murray 2006.01.24

http://groups.yahoo.com/group/aspartameNM/message/1279
all three aspartame metabolites harm human erythrocyte [red blood cell]
membrane enzyme activity, KH Schulpis et al, two studies in 2005,
Athens, Greece, 2005.12.14: 2004 research review, RL Blaylock:
Murray 2006.01.14

http://groups.yahoo.com/group/aspartameNM/message/939
aspartame (aspartic acid, phenylalanine) binding to DNA:
Karikas July 1998: Murray 2003.01.05 rmforall
Karikas GA, Schulpis KH, Reclos GJ, Kokotos G
Measurement of molecular interaction of aspartame and
its metabolites with DNA. Clin Biochem 1998 Jul; 31(5): 405-7.
Dept. of Chemistry, University of Athens, Greece
http://www.chem.uoa.gr gkokotos@...;
K.H. Schulpis inchildh@...; G.J. Reclos reklos@...;

http://groups.yahoo.com/group/aspartameNM/message/1271
combining aspartame and quinoline yellow, or MSG and brilliant blue,
harms nerve cells, eminent C. Vyvyan Howard et al, 2005
education.guardian.co.uk, Felicity Lawrence: Murray 2005.12.21

http://groups.yahoo.com/group/aspartameNM/message/925
aspartame puts formaldehyde adducts into tissues, Part 1/2
full text Trocho & Alemany 1998.06.26
Universitat Auṭnoma de Barcelona : Murray 2002.12.22

http://groups.yahoo.com/group/aspartameNM/message/1250
aspartame causes cancer in rats at levels approved for humans,
Morando Soffritti et al, Ramazzini Foundation, Italy &
National Toxicology Program
of National Institute of Environmental Health Sciences
2005.11.17 Env. Health Pers. 35 pages: Murray

http://groups.yahoo.com/group/aspartameNM/message/1106
hangover research relevant to toxicity of 11% methanol in aspartame
(formaldehyde, formic acid): Calder I (full text): Jones AW:
Murray 2004.08.05 rmforall

Since no adaquate data has ever been published on the exact disposition
of toxic metabolites in specific tissues in humans of the 11% methanol
component of aspartame, the many studies on morning-after hangover
from the methanol impurity in alcohol drinks are the main available
resource to date.

Jones AW (1987) found next-morning hangover from red wine with
100 to 150 mg methanol
(9.5% w/v ethanol, 100 mg/l methanol, 0.01%,
one part in ten thousand).

http://groups.yahoo.com/group/aspartameNM/message/870
Aspartame: Methanol and the Public Interest 1984: Monte:
Murray 2002.09.23

Humans suffer "toxic syndrome" (54) at a minimum lethal dose
of <1 gm/kg, much less than that of monkeys, 3-6 g/kg (42, 59).

The minimum lethal dose of methanol
in the rat, rabbit, and dog is 9.5, 7.0 , and 8.0 g/kg, respectively (43);
ethyl alcohol is more toxic than methanol to these test animals (43)."


http://groups.yahoo.com/group/aspartameNM/message/909
testable theory of MCS type diseases, vicious cycle of nitric oxide &
peroxynitrite: MSG: formaldehyde-methanol-aspartame:
Martin L. Pall: Murray: 2002.12.09 rmforall

Environ Health Perspect. 2003 Sep; 111(12): 1461-4.
Elevated nitric oxide/peroxynitrite theory of multiple chemical sensitivity:
central role of N-methyl-D-aspartate receptors
in the sensitivity mechanism. Pall ML.
School of Molecular Biosciences, 301 Abelson Hall, Washington State
University, Pullman, WA 99164, USA. martin_pall@...

The elevated nitric oxide/peroxynitrite and the neural sensitization
theories of multiple chemical sensitivity (MCS) are extended here
to propose a central mechanism for the exquisite sensitivity
to organic solvents apparently induced
by previous chemical exposure in MCS.
This mechanism is centered
on the activation of N-methyl-D-aspartate (NMDA) receptors
by organic solvents producing elevated nitric oxide and
peroxynitrite, leading in turn to increased stimulating of and
hypersensitivity of NMDA receptors.
In this way, organic solvent exposure may produce
progressive sensitivity to organic solvents.
Pesticides such as organophosphates and carbamates
may act via muscarinic stimulation to produce a similar biochemical
and sensitivity response.
Accessory mechanisms of sensitivity may involve
both increased blood-brain barrier permeability,
induced by peroxynitrite,
and cytochrome P450 inhibition by nitric oxide.
The NMDA hyperactivity/hypersensitivity and excessive nitric
oxide/peroxynitrite view of MCS provides answers
to many of the most puzzling aspects of MCS
while building on previous studies and views of this condition.
PMID: 12948884

Prof. Pall describes processes by which an initial trigger exposure,
such as carbon monoxide or formaldehyde,
can generate hypersensitivity to many substances.
He himself had recovered from a sudden, debilitating attack of
multiple chemical sensitivity in June/July 1997.

http://groups.yahoo.com/group/aspartameNM/message/1055
hormesis: possible benefits of low-level aspartame
(methanol, formaldehyde) use: Calabrese: Soffritti:
Murray 2004.03.11

http://groups.yahoo.com/group/aspartameNM/message/1056
disorders of NMDA glutamate receptors in brain range
from high activity (MCS, CF, PTSD, FM, from carbon monoxide
or formaldehyde (methanol, aspartame)-- Pall)
to low activity (schizophrenia-- Coyle, Goff, Javitts):
Murray 2004.03.13
*******************************************************