Fluoride Deposition in the Aged Human Pineal Gland

[Guest guest]

It has been very difficult to find " scientific truth " for a very long

time now. Most of it has usually come from countries who do not

benefit directly from the the sale of some " product " involved. In the

USA it is especially hard to find. In the modern day climate what

passes for " scientific truth " depends on if you are pitching or

catching or selling or buying.

 

 

 

http://www.icnr.com/jluke/FluorideDeposition.htm

 

 

 

 

Fluoride Deposition in the Aged Human Pineal Gland

Jennifer Luke

School of Biological Sciences, University of Surrey, Guildford, UK

Department of Obstetrics and Gynaecology, The Royal London Hospital

 

 

 

Original Paper

Caries Res 2991;35:125-128

 

2001 S. Karger AG, Basel

Presented by permission of the author.

 

Key Words

Calcium, Distribution, Fluoride, Human pineal gland, Hydroxyapatite,

Pineal concretions

 

Abstract

The purpose was to discover whether fluoride (F) accumulates in the

aged human pineal gland. The aims were to determine (a)

F-concentrations of the pineal gland (wet), corresponding muscle (wet)

and bone (ash); (b) calcium-concentration of the pineal. Pineal,

muscle and bone were dissected from 11 aged cadavers and assayed for F

using the HMDS-facilitated diffusion, F-ionspecific electrode method.

Pineal calcium was determined using atomic absorption spectroscopy.

Pineal and muscle contained 297±257 and 0.5±0.4 mg F/kg wet weight,

respectively; bone contained 2,037±1,095 mg F/kg ash weight. The

pineal contained 16,000±11,070 mg Ca/kg wet weight. There was a

positive correlation between pineal F and pineal Ca (r = 0.73, p<0.02)

but no correlation between pineal F and bone F. By old age, the pineal

gland has readily accumulated F and its F/Ca ratio is higher than bone.

 

The pineal gland is a small organ situated near the centre of the

brain. It is intimately related to the third ventricle. It is composed

of pinealocytes and neuroglial cells amongst which ramifies a rich

network of capillaries and postganglionic nerve fibres. The pineal

gland is a mineralizing tissue. Its calcified concretions range from a

few micrometres to several millimetres in diameter. The larger ones

are identifiable on skull X-rays, cranial CT and MRI scans. The

concretions are composed of hydroxyapatite (HA) [Angervall et al.,

1958; Earle, 1965; Mabie and Wallace, 1974; Galliani et al., 1990;

Bocchi and Valdre, 1993] whose chemical composition, morphology, and

unit cell dimensions are similar to HA in bone and teeth [Mabie and

Wallace, 1974; Bocchi and Valdre, 1993]. The pineal (calcified and

uncalcified) has a high trace element content (zinc, iron, manganese,

magnesium, strontium and copper) in humans [Krstic, 1976; Michotte et

al., 1977] and in rats [Humbert and Pévet, 1991, 1996]. Michotte et

al. [1977] suggested that, within the pineal, there are areas which

are heavily loaded with calcium and which attract trace elements, even

though these calcium-rich areas are not yet identifiable as

concretions. Calcium is distributed throughout the pinealocytes: in

the mitochondria, Golgi apparatus, cytoplasm, and nucleus [Krstic,

1976, 1995; Welsh, 1984; Pizarro et al., 1989, Lewczuk et al., 1994].

 

Fluoride does not accumulate in brain. Of all tissues, brain has the

lowest fluoride concentration [Jenkins, 1991; Whitford, 1996;

Ekstrand, 1996]. It is generally agreed that the blood-brain barrier

restricts the passage of fluoride into the central nervous system. The

human pineal gland is outside the blood-brain barrier [Arendt, 1995].

It is one of a few unique regions in the brain (all midline structures

bordering the third and fourth ventricles) where the blood-brain

barrier is weak. Cells in these regions require direct and unimpeded

contact with blood [Rapoport, 1976]. Therefore, pinealocytes have free

access to fluoride in the bloodstream. This fact, coupled with the

presence of HA, suggest that the pineal gland may sequester fluoride

from the bloodstream.

 

The purpose of this study was to discover whether fluoride accumulates

in the aged pineal gland. Its objectives were to determine (a) the

fluoride concentrations of the pineal gland (wet), corresponding

muscle (wet) and bone (ash); (b) the pineal concentrations of calcium

and HA.

 

Materials and Methods

The pineal glands and corresponding bone and muscle samples were

dissected from I I aged cadavers (7 females and 4 males) in the

Anatomy Department, UCL. The mean age was 82 years (range 70-100).

 

Preparation of the Samples

The pineal glands were blotted dry with tissue paper, weighed to the

nearest milligram, homogenized in I ml double-distilled water using an

agate pestle and mortar and sonicated for 10 min. Each pineal was

divided into two portions that were analysed separately. Muscle

samples weighing about 100 mg were treated likewise. Bone samples were

cleaned of any adherent soft tissue with a razor blade, dried

overnight at 110'C in an oven, and ashed (in porcelain crucibles with

no fixatives) at 550-600°C in a muffle furnace for 8 h. Bone ash was

pulverized into a fine powder using an agate pestle and mortar. Bone

solutions were made by dissolving known weights of bone ash in 3 ml 2

M HC104. Bone solutions were analysed for fluoride in replicates of

six and the mean fluoride concentrations in bone were calculated.

 

Determination of the Fluoride Concentrations

The homogenized pineal, homogenized muscle and bone solutions were

assayed for fluoride using the HMDS-facilitated diffusion,

F-ion-specific electrode method originally described by Taves [19681

and modified by Whitford and Reynolds [1979]. The protocol was further

modified for use with the pineal glands. The concentration and volume

of the base trap were increased to 0.5 M NaOH and 100 µl,

respectively; the strength of the acetate buffer was increased to 50

µl 2 M acetic acid and the volume of the analysed solution was

adjusted to 150 µl with double-distilled water. Diffusion time for

pineal and muscle was 3 days; for bone 18 h.

 

Standards: pineal: 1,000, 2,500 and 5,000 nmol F; muscle: 5, 50 and

500 nmol F; bone: 10, 50, 100 and 500 nmol F.

 

Determination of the Concentration of Total Calcium in the Human

Pineal Gland

The acid digests, which remained in the Petri dishes following the

separation of fluoride from the pineal glands, were wet-acid ashed to

decompose the organic component. The acid digests were placed in clean

glass tubes and I ml conc. HN03 was added. The tubes were heated

slowly to 50°C and maintained at 50°C for 30 min in a fume cupboard.

The procedure was repeated using I ml 60% HC104. Two millilitre of

double-distilled water was added to each tube and the volumes were

measured. Calcium concentration was determined using atomic absorption

spectroscopy.

 

Statistical Methods

Results were expressed as means ± SD. Differences between the groups

were tested for significance using unpaired Student's t-test.

Differences were regarded as statistically significant when p<0.05.

 

Pearson's correlation coefficient was used to test association between

pineal fluoride and pineal calcium; and pineal fluoride and bone fluoride.

 

 

Fig. 1. The relationship between the calcium and fluoride contents of

ten aged human pineal glands.

 

Results

The aged pineal gland weighed 112±52 mg (56-198 mg). Pineal had a

significantly higher F concentration than muscle: 297 ± 257 (14-875)

vs. 0.5 ± 0.4 (0.2-1.5) mg F/kg wet weight (p<0.001). Bone contained

2,037± 1,095 (838-3,711) mg F/kg ash weight. The mean coefficient of

variation between the replicates of F contents of bone solutions was

2.5 ± 1. 1 %. There was no correlation between pineal F and bone F.

The pineal gland contained 16,000±11,070 (4,600-37,250) mg Ca/kg wet

weight. Pineal fluoride and pineal calcium were directly correlated: r

= 0.73, p < 0.02, n = 10, slope= 0.02 (fig. 1). Assuming

stoichiometric HA, the pineal contained an estimated 40,000 ± 27,700

mg HA/kg wet weight (11,600-93,200 mg HA/kg). The estimated F

concentration of pineal HA was 9,000 ± 7,800 mg/kg (650-21,800 mg/kg).

Figure 2 shows that the F/calcium ratio was higher in pineal HA than

in corresponding bone HA.

 

Discussion

This study has added new knowledge on the fate and distribution of

fluoride in the body. It has shown for the first time that fluoride

readily accumulates in the human pineal gland although there was

considerable inter-individual variation (14-875 mg F/kg). By old age,

the average pineal gland contains about the same amount of fluoride as

teeth (300 mg F/kg) since dentine and whole enamel contain 300 and 100

mg F/kg, respectively [Newbrun, 1986]. Unlike brain capillaries,

pineal capillaries allow the free passage of fluoride through the

endothelium. If there had been a bloodbrain barrier in the pineal, it

would have prevented the passage of fluoride into the pinealocytes and

the pineal fluoride content would have been similar to or lower than

muscle. This was obviously not the case: the fluoride concentration of

the pineal was significantly higher (p<0.001) than muscle. The high

fluoride levels in the pineal are presumably due to the large surface

area of the HA crystallites both intra- and extracellularly. In

addition, the pineal has a profuse blood flow and high capillary

density; pineal blood flow (4 ml/min/g) is second only to the kidney

[Arendt, 19951.

 

The extent of pineal calcification also varied between individuals:

ranging from 4,600 to 37,250 mg Ca/kg wet weight. One of the aged

pineals had very little precipitation. This supports the age

independence of pineal calcification and agrees with previous studies

[Cooper, 1932; Arieti, 1954; Tapp and Huxley, 197 1; Hasegawa et aL,

1987; Galliani et al., 1990]. The estimated fluoride concentration of

pineal HA was 9,000 ± 7,800 mg/kg. The F/Ca ratio was higher in pineal

HA than in corresponding bone (fig. 2). The extremely high level of

substitution in the crystal structure of pineal HA by fluoride

illustrates the readiness with which fluoride replaces the hydroxyl

ion in the HA crystal. By old age, pineal HA has a higher fluoride

content than other biological apatites. Unlike pineal concentrations

of magnesium, manganese, zinc and copper, which, although very high,

were generally within the limits found in bone and teeth [Michotte et

al., 1977].

 

 

Fig. 2. A comparison of the F/Ca ratio in aged pineal HA and

corresponding bone ash (µg F/100 µg calcium). Calcium content of bone

determined stoichiometrically.

 

There was no corTelation between pineal fluoride and bone fluoride.

Therefore, unlike bone, pineal fluoride concentrations are not

indicators of long-term fluoride exposure and body burden. Pineal

fluoride, however, was significantly correlated with pineal calcium.

 

The methodology used in this project was accurate because the F values

obtained for bone and muscle agreed with literature values. For

example, the mean fluoride concentration of bone from elderly subjects

was 2,000 mg/kg ash weight which agrees with previous studies using

bone from subjects of a similar age [Ebie et al., 1992; Charen et al.,

1979; Zipkin et al., 1958]. In this study, muscle contained 0.5 mg

F/kg wet weight, a typical fluoride concentration for soft tissue

[WHO, 1984]. The pineal and bone were treated differently during

sample preparation (the pineal was wet-acid ashed, bone was dry ashed)

which may somewhat obscure a direct comparison of the fluoride

contents of pineal HA and bone. However, it is unlikely that there

would be a significant analytical error.

 

In conclusion, this study presented evidence that fluoride readily

accumulates in the aged pineal. Fluoride may also accumulate in a

child's pineal because significant amounts of calcification have been

demonstrated in the pineals from young children [Cooper, 1932;

Wurtman, 1968; Kerényi and Sarkar, 1968; Tapp and Huxley, 197 1;

Doskocil, 1984]. In fact, calcification of the developing enamel

organs and the pineal gland occur concurrently. If fluoride does

accumulate in the child's pineal (this needs verification), the

pinealocytes will be exposed to relatively high local concentrations

of fluoride. This could affect pineal metabolism in much the same way

that high local concentrations of fluoride in the developing enamel

organ affect ameloblast function. Research is presently underway to

discover whether fluoride affects pineal physiology during childhood:

specifically pineal synthesis of melatonin.

 

Acknowledgements

I acknowledge the Anatomy Deptartment, UCL, for providing the pineal

glands; Prof. Gary Whitford, Medical College of Georgia, USA, and

Gordon Hartman, University of Surrey, for their assistance with the

fluoride analysis; Nicholas Porter, The Royal Surrey County Hospital,

Guildford, for help with the determination of pineal-calcium

concentrations. The Colt Foundation, The Heinz and Anna Kroch

Foundation and The New Moorgate Trust funded the research. References

 

1. Angervall L, Berger S, Röckert H: A microradiographic and X-ray

crystallographic study of calcium in the pineal body and intracranial

tumours. Acta Pathol Microbiol Scand 1958;44: 113-119.

2. Arendt J: Melatonin and the Mammalian Pineal Gland, ed 1.

London. Chapman & Hall, 1995, p 17.

3. Arieti S: The pineal gland in old age. J Neuropathol Exp Neurol

1954; 13:482-49 1.

4. Bocchi G, Valdre G: Physical, chemical, and mineralogical

characterization of carbonate-hydroxyapatite concretions of the human

pineal gland. J Inorg Biochem 1993;49:209-220.

5. Charen J, Taves DR, Stamm JW, Parkins FM: Bone fluoride

concentrations associated with fluoridated drinking water. Calcif Tiss

Int 1979;27: 95-99.

6. Cooper ERA: The human pineal gland and pineal cysts. J Anat

(Lond) 1932;67:28-46.

7. Doskocil M: Development of concrements in the human pineal body.

Folia Morphol (Praha) 1984;32:16-26.

8. Earle KM: X-ray diffraction and other studies of the calcareous

deposits in human pineal glands. J Neuropathol Exp Neurol 1965;4: 108-118.

9. Ebie DM, Deaton TG, Wilson FC, Bawden JW: Fluoride

concentrations in human and rat bone. J Public Health Dent

1992;52:288-29).

10. Ekstrand J: Fluoride metabolism; in Fejerskov 0, Ekstrand J,

Burt B (eds): Fluorides in Dentistry. Munksgaard, Copenhagen, 1996, pp

55-68.

11. Galliani 1, Falcieri E, Giangaspero F, Valdre G, Mongiorgi R: A

preliminary study of human pineal gland concretions: Structural and

chemical analysis. Boll Soc Ital Biol Sper 1990;66: 615-622.

12. Hasegawa A, Ohtsubo K, Mori W: Pineal gland in old age;

quantitative and qualitative morphological study of 168 human autopsy

cases. Brain Res 1987;409:343-349.

13. Humbert W, Pévet P: Calcium content and concretions of pineal

glands of young and old rats, Cell Tissue Res 199 1;263:593-596.

14. Humbert W, Pévet P: Electron probe x-ray microanalysis of the

elemental composition of the pineal gland of young adults and aged

rats. J Pineal Res 1996;20:39-44.

15. Jenkins GN: Physiology of fluoride; in Murray JJ, Rugg-Gunn AJ,

Jenkins GN (eds): Fluorides in Caries Prevention, ed 3. London,

Butterworth-Heinemann, 199 1, pp 262-294.

16. Kerényi NA, Sarkar K: The postnatal transformation of the pineal

gland. Acta Morphol Acad Sci Hung 1968; 16:223-236.

17. Krstic R: A combined scanning and transmission electron

microscopic study and electron probe microanalysis of human pineal

acervuli. Cell Tiss Res 1976; 174:129-137.

18. Krstic R: Ultracytochemical localization of calcium in the

superficial pineal gland of the Mongolian gerbil. J Pineal Res

1985;2:21-37.

19. Lewczuk B, Przybylska B, Wyrzykowski Z: Distribution of

calcified concretions and calcium ions in the pig pineal gland. Folia

Histochem Cytobiol 1994;32:243-249.

20. Mabie CP, Wallace BM: Optical, physical and chemical properties

of pineal gland calcifications. Calcif Tissue Res 1974; 16:59-7 1.

21. Michotte Y, Lowenthal A, Knaepen L, Collard M, Massart DL: A

morphological and chemical study of calcification of the pineal gland.

J Neurol 1977;215:209-219.

22. Newbrun E: Fluorides and Dental Caries, ed 3. Springfield,

Thomas, 1986.

23. Pizarro MDL, Gil JAL, Vasallo JL, Munoz Barragan L: Distribution

of calcium in the pineal glands of normal rats. Adv Pineal Res 1989;

3:39-42.

24. Rapoport SI: Blood-Brain Barrier in Physiology and Medicine. New

York, Raven Press, 1976, p 77-78.

25. Tapp E, Huxley M: The weight and degree of calcification of the

pineal gland. J Pathol 197 1; 105:31-39.

26. Taves DR: Separation of F by rapid diffusion using

hexamethyldisiloxane. Talanta 1968;15:969-974.

27. Welsh MG: Cytochemical analysis of calcium distribution in the

superficial pineal gland of the Mongolian gerbil. J Pineal Res

1984;1:305-316.

28. Whitford GM: The Metabolism and Toxicity of Fluoride. Monogr

Oral Sci, 16, ed 2. Basel, Karger, 1996.

29. Whitford GM, Reynolds KE: Plasma and developing enamel fluoride

concentrations during chronic acid-base disturbances. J Dent Res

1979;58:2058-2065.

30. WHO: Fluorine and Fluorides. Environmental Health Criteria 36.

Geneva, WHO, 1984.

31. Wurtman RJ: The pineal gland; in Endocrine Pathology. Baltimore,

Williams & Wilkins, 1968, pp 117-132.

32. Zipkin 1, McClure FJ, Leone NC, Lee WA: Fluoride deposition in

human bones after prolonged ingestion of fluoride in drinking water.

US Public Health Rep 1958;73:732-740.