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Study on changing concentration of TNT and some cytokines in workers occupationally exposed to tnt after non-specific detoxification treatment

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Subjects and methods: A clinical trial study was conducted on 70 workers in some military factories with the evidence of trinitrotoluene (TNT) in their blood from October 2014 to April 2015. The non-specific detoxification method had been applied according to the scientific principle of the Hubbard method for 3 weeks.

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STUDY ON CHANGING CONCENTRATION OF TNT AND SOME CYTOKINES IN WORKERS OCCUPATIONALLY EXPOSED TO TNT AFTER NON-SPECIFIC DETOXIFICATION TREATMENT

Nguyen Kien Cuong*; Nguyen Phuc Thai*; Do Phuong Huong**

Nguyen Xuan Kien***; Nguyen Minh Phuong****

SUMMARY

Subjects and methods: A clinical trial study was conducted on 70 workers in some military factories with the evidence of trinitrotoluene (TNT) in their blood from October 2014 to April

2015 The non-specific detoxification method had been applied according to the scientific principle of the Hubbard method for 3 weeks Results : Mean TNT concentration before

treatment was 0.150 (µg/mL) and reduced sharply down to 0.0154 (µg/mL) after treatment The difference was statistically significant (p < 0.05) Mean concentration of erythropoietin (EPO) pre-treatment was 676.629 (pg/mL) and rose 792.44 (pg/mL) after the treatment (p < 0.05) Mean erythropoietin receptor (EPOR) before treatment was 4574.23 (pg/mL) and increased up

to 5982.89 (pg/mL) after treatment, the difference was statistically significant (p < 0.05) Mean concentration of interferon gamma (IFN-γ) was 395.46 (pg/mL) before treatment and reached 621.12 (pg/mL) after the detoxification (p < 0.05)

* Keywords: TNT; Detoxification; Occupational exposure; Women; Erythropoietin; Erythropoietin receptor; Interferon gamma

INTRODUCTION

Trinitrotoluene has been widely used in

the defense with the large number of

workers directly exposed to TNT Until now,

there is no specific detoxification treatment

for TNT TNT not only damages the blood

cells in bone marrow and periferal [11] but

also inhibites the body's immune system

[4] Prophylaxis of occupational TNT toxicity

plays an important role in ensuring the

health of workers exposed to TNT

L.Ron Hubbard has successfully

researched and applied non-specific

detoxification method to a variety of lipid-soluble chemicals such as chlorobenzo-cyclic hydrocarbons (PCBs…), heavy metals (lead, mercury), etc However, this method has not been conducted for the detoxification

of TNT, one kind of lipid soluble chemicals

If the nonspecific detoxification for the TNT workers is effective, how will it affect some cytokines that plays an important role in hematopoietic and immune stimulation system? From these reasons,

we conducted a study with a view to:

Evaluating TNT, EPO, EPOR and IFN-γ

* Mlitary Institute of Preventive Medicine

** 103 Military Hospital

*** Military Medicine Department

**** Vietnam Military Medical University

Corresponding author: Nguyen Minh Phuong (phuongk21@gmail.com)

Date received: 06/07/2017 Date accepted: 18/08/2017

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level after treatment with non-specific

detoxification which applied Hubbard's

scientific principles The results will

contribute to assess therapeutic effects of

non-specific detoxification for TNT exposure

workers

SUBJECTS AND METHODS

1 Subjects

- 70 workers work for some military

plants, directly exposed to TNT, with the

evidence of TNT in their blood

- Workers had been selected with no

chronic diseases such as diabetes, high

blood pressure, etc… and no acute diseases

that needs to be cured

- Volunteer to participate in this research

- Workers were treated with

non-specific detoxification at Detox Center,

Military Hospital 103 for 3 weeks The

research was conducted from October,

2014 to April, 2015

2 Methods

- Methodology of clinical trial, evaluation pre-and post-intervention, with the permission from the Ethics Board of

103 Military Hospital

- Assessment of changes in TNT, EPO, EPOR and IFN-γ levels before and after non-specific detoxification treatment Non-specific detoxification process consists of 3 following key contents: Running exercise in 30 minutes before the dried sauna for 2.5 hours right after exercise All the workers had been supplied niacin and mineral salt, water, vitamins A, D, C, E, B and vegetable oil (unsaturated fatty acids)

- Data had been analyzed by SPSS software Comparison of mean values of TNT, EPO, EPOR and IFN-γ before and after treatment

RESULTS AND DISCUSSION

1 Characteristics of the subjects

Table 1: Characteristics of the research subjects

Subjects (n = 70) Characteristics

- The average age of the participants was 42.02 ± 6.26 They are all exposed to TNT and they have no signs of occupational TNT toxicity In fact, they can be considered as healthy people

- The proportion of female and male workers accounted for 51.4%, and 48.6%,

respectively The rate of women in our study is basically consistent with the proportion

of women labour structure in the Red River Delta (50.6%) [1]

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2 Changes in blood TNT concentration before and after treatment

Table 2: Blood TNT concentration pre- and post-treatment

Blood TNT

(µg/mL)

Before treatment (n = 70)

After treatment (n = 70)

The plasma TNT level before treatment

was 0.150 (µg/mL), after detoxification

TNT concentration reduced sharply to

0.0154 (µg/mL) The difference was

statistically significant (p < 0.05)

The highest TNT level before treatment

was 0.51 µg/mL and after treatment was

0.16 µg/mL

The results showed that the effects of

detoxification for TNT were well-activated

According to the science principles of

Hubbard methodology, toxins accumulate in

the lipid organization were excreted from

the body by doing exercise, sauna and

mobilizing lipid metabolism, which

accelerates TNT clearance more rapidly

than TNT's natural metabolic pathway

Niacin (vitamin B3) boosts the release of free fatty acids, which stimulates the release of accumulated xenobiotics in the fats and allows them to be excreted through the skin [6]

Reports on TNT dynamics in humans are still very rare It was observed that after 17 days of leaving the working environment, the presence of

dinitroaminotoluene (one metabolic of TNT) was still observed in the urine at 0.06 mg/L (Woolen BH et al, 1986) This indicated that TNT and its metabolites eliminated relatively slow in humans [12]

The results of our study revealed that TNT still accumulated in the body after 3 weeks

of leaving the working environment

3 Changes in some cytokines before and after treatment

Table 3: EPO changes before and after the treatment

(n = 70)

After treatment (n = 70)

p < 0.05

Mean EPO levels before and after

treatment were 676.629 pg/mL and

792.44 pg/mL, respectively The difference

was statistically significant (p < 0.05)

The increase in EPO level after

exercise has been well documented

Which mechanism that regulates and mobilizes hematopoietic cells after exercise are still unknown clearly Kidney is a major EPO production site, EPO deficiency

is commonly observed in people with anemia such as renal failure Experimental

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study by Baker et al (2011) showed that in

trained muscle groups, there was an

increase in the expression level of the

EPO up to 93% [2]

In bone marrow, EPO binds to its

receptor (EPOR) which is expressed on

the surface of erythroid progenitors, where

it stimulates the proliferation and suppresses

the apoptosis of newly formed cells

Under hypoxia, EPO gene transcription is

activated, resulting in an increase in EPO

level in the circulation, afterward EPO

binds to its specific receptor on the

surface of erythroid precursors in the

bone marrow, initiate the JAK2 (Janus

kiansse 2) signal pathway, and increase

the number of red blood cells

EPO is known to play a role in blood

vessels protection by acting on endothelial

cells and smooth muscle cells in blood

vessels through the mechanism of inhibiting apoptosis or by stimulating the production Nitric oxide and vasodilatation enhancement [10] From our point of view, due to the vasodilatation effect that was made by the combination of EPO and niacin during the treatment, therefore enhance the removal of toxins from the body It can be said that the enhancement

of EPO after the detoxification provides positive protective effects

We suggested that an increasing EPO after treatment is possibly due to the effects

of sauna, high temperature increasing vassolidation, leads to increase blood flow through the skin Therefore, it can reduce oxygen-carrying blood flow to organs especially the kidneys, which activates the kidneys to boost the production of EPO by reverse-regulating mechanism

Table 4: EPOR changes before and after the treatment

(n = 70)

After treatment (n = 70)

The average EPOR pre-treatment level was 4574.23 pg/mL, increased to 5982.89 pg/mL

after the treatment, statistically significant differences (p < 0.05)

EPOR is produced by lack of oxygen and leads to an increase in EPO sensitivities

The interaction between EPO and EPOR creates biological effects To the best of our

knowledge, the increase in the concentration of EPO and EPO receptors synergizes

and promotes the biological effect of EPO

Table 5: IFN-γ changes before and after the treatment

(n = 70)

After treatment (n = 70)

p < 0.05

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Mean IFN-γ level after treatment is

higher than that before treatment Prior to

treatment, the average IFN-γ level was

395.46 pg/mL After treatment, mean IFN-γ

level rose to 621.12 pg/mL, the difference

was statistically significant (p < 0.05)

The increase in IFN-γ level after

non-specific detoxification may be due to the

use of some vitamins, including the role of

vitamin E and vitamin A during treatment

Vitamin E has been shown to increase the

CD4/CD8 ratio Moreover, rising IFN-γ

production has been demonstrated in

patients with colorectal cancer using

vitamin E to enhance immunity [8]

Vitamin E has been known as

anti-inflammatory and antioxidant abilities, the

use of vitamin E in treatment is quite

widespread and common Because of the

anti-free radical effect, vitamin E plays a

significant role in treating many diseases

of various organs in the body [9]

In our opinion, the increase in IFN-γ

level after non-specific treatment is not

only the role of vitamins, but also the role

of heat stimulation due to sauna and the

effect of exercise In experiment, it was

found that hyperthermia increased the

ability of inhibition tumor growth, enhancing

the survivor time of animal with experimental

tumors [5] After a moderate exercise, M

Baum (1997) reported an increase in IFN-γ

level among participants, an IFN-γ

concentration of 974 pg/mL (before

practicing) increased to 1,450 pg/mL after

24 hours of doing exercise

Therefore, from the findings of our study

on changes in IFN-γ levels before and

after non-specific detoxification based on

the Hubbard method, it can be said that

non-specific detoxification applied Hubbard method including combined exercises, sauna and vitamins supplementation is effective to stimulate immune system and enhance the body's resistance

CONCLUSION

TNT level before treatment was 0.150 µg/mL, and reduced sharply to

0.0154 µg/mL after treatment The difference was statistically significant (p < 0.05)

Mean concentration of EPO pre-treatment were 676.629 pg/mL and rosed

up to 792.44 pg/mL after the treatment (p < 0.05)

Mean EPOR before treatment was 4574.23 pg/mL and increased to 5982.89 pg/mL after treatment, the difference was statistically significant (p < 0.05)

Mean concentration of IFN-γ were 395.46 pg/mL before treatment and increased up to 621.12 pg/mL after the detoxification (p < 0.05)

RECOMMENDATIONS

Step by step to do research on non-specific TNT detoxification method at military medicine at baseline in military factories, workers exposed to TNT should

be periodically treated by non-specific detoxification to protect and promote their

health

REFERENCES

1 Tổng cục Thống kê Báo cáo điều tra lao

động và việc làm quý 2 năm 2014 2014, tr.4

2 Baker J.M, De Lisio M, Parise G

medullary hematopoiesis Faseb J 2011, 25 (12), pp.4348-4357

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3 Baum M., Muller-Steinhardt M, Liesen H

et al Moderate and exhaustive endurance

exercise influences the interferon-gamma

levels in whole-blood culture supernatants

Eur J Appl Physiol Occup Physiol 1997, 76

(2), pp.165-169

4 B runs - Nagel D, Scheffer S et al Effect

of 2,4,6-trinitrotoluene and its metabolites on

human monocytes Environmental Science &

Technology.1999, 33 (15), pp 2566-2570

5 Hattori T, Kokura S, Okuda T et al

Antitumor effect of whole body hyperthermia

subcutaneous tumor model of colon cancer

Int J Hyperthermia 2007, 23 (7), pp 591-598

6 Jonathan E Prousky N, MSc Niacin for

detoxification: A Little-known therapeutic use

Journal of Orthomolecular Medicine 2011, 26 (2)

7 Lee F.S, Percy M.J The HIF pathway

and erythrocytosis Annu Rev Pathol 2011, 6,

pp.165-192

8 Malmberg K.J, Lenkei R, Petersson M et

al A short-term dietary supplementation of

high doses of vitamin E increases T helper 1 cytokine production in patients with advanced colorectal cancer Clin Cancer Res 2002, 8 (6), pp.1772-1778

9 Peh H.Y, Tan W.S, Liao W et al Vitamin

E therapy beyond cancer: Tocopherol versus tocotrienol Pharmacol Ther 2016, 162, pp.152-69

10 Santhanam A.V, d'Uscio L.V, Katusic Z.S Cardiovascular effects of erythropoietin

an update Adv Pharmacol 2010, 60,

pp.257-285

11 U.S Department of Health and Human Services Toxicological profile for 2,4,6 -

Trinitrotoluene Agency for Toxic Substances and Disease Registry 1995

12 Woollen B.H, Hall M.G, Craig R et al

trinitrotoluene: Assessment of occupational absorption during manufacture of explosives

Br J Ind Med 1986, 43 (7), pp.465-473

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