Screening for Early detection of subclinical hypothyroidism: prognostic value of 3D-NLS bio resonance feedback diagnostic

Mohammad Ebrahimi ()21^,2, Vera Ivanovna Nesterova³, Vladimir Igorevich Nesterov³.,

1.The Research Center for New Technologies in Life Science Engineering, Institute of Bio-signal and Immunculus, Tehran University, Tehran, Iran

2.The Medical Scientific Center for Development of Bioelectric and Bio-feedback Technologies, Tehran, Iran

3.The Institute of Practical Psychophysics, 2, 1st Proizvodstvennaya street, 644001 Omsk, Russia.,

 Key words: Sub-hypothyroidism, Screening, TSH, Ultrasound, 3D-NLS bio resonance feed- back diagnostic, Spectral-entropy-analysis.

Abbreviation:

SCH- subclinical hypothyroidism

US – ultrasonography

3D NLS BR- 3D-Nonlinear Bio feedback bioresoance system

SEA – spectral-entropy analysis

Abstract: Hypothyroidism is a commonly seen condition in medical practice. Subclinical hypothyroidism can progress to overt hypothyroidism. Hypothyroidism as a chronic disease has long and variable preclinical phases. The preclinical phase is that portion of the disease’s natural history during which the disease is potentially detectable, but unrecognized.  Screening of individuals for sub-clinical thyroids in important, because this condition can lead to overt hypothyroidism. Screening has two major objectives. One is the early detection of the disease at a point when treatment is more effective and less expensive. And the second objective in screening is to identify risk factors that render an individual at a higher risk for developing a disease, with the goal of modifying the risk factors to prevent or reduce the disease.

TSH is the accepted first-line screening test for the diagnosis of the majority of patients suspected of having hypothyroidism or hyperthyroidism and is measured by automated immunoassays.

Serum TSH measurements may yield misleading results for individuals with changing levels of thyroid hormones and the secretion and action of TSH affected by many factors such as non-thyroidal disorder, immune system disturbance, infections, different medications, aging and other factors. Thyroid ultrasonography (US) has established itself as a common and valuable tool in the evaluation and management of thyroid disorder, but US also has some limitation is scanning of thyroid gland.  But, neither the TSH serum level, nor the ultrasound method can meet both criteria in screening.

In this study, biophysical markers were used for evaluation of thyroid dysfunction. The NLS-3D bio-feedback bioresoance system was used for evaluation 3 biophysical spectral-entropy parameters. Analysis data extracted by using 3D NLS bioresonance feedback system showed that Spectral Entropy Analysis (SEA) parameters are valuable markers for early detection of thyroid-related disorders and can be used in screening of asymptomatic patients.

Introduction:

Hypothyroidism is a decreased functioning of thyroid gland and common conditions among chronic diseases and the most common disorder arising from hormone deficiency. It has lifelong effects on health [1-2].

According to the time of onset, it is divided in congenital and acquired, according to the level of endocrine dysfunction in primary and secondary or central and according to the severity in severe or clinical and mild or subclinical hypothyroidism (SCH). Hypothyroidism is a commonly seen condition. The most common cause of primary hypothyroidism is thyroiditis due to antithyroid antibodies which results from under-secretion of thyroid hormone, a condition called “Hashimoto’s thyroiditis.” and secondary hypothyroidism is caused by lack of TSH production from the pituitary [3-4].

The distinction between subclinical and clinical hypothyroidism is of major significance, as in clinical hypothyroidism symptoms are more severe -even coma may occur, while in subclinical hypothyroidism, symptoms are less serious and may even be absent. Subclinical hypothyroidism can progress to overt hypothyroidism, especially if antithyroid antibodies are present, and has been associated with adverse metabolic, cardiovascular, reproductive, maternal-fetal, neuromuscular, and cognitive abnormalities and lower quality of life [5-7]. Persons with subclinical thyroid dysfunction have been found to have a higher mean body mass index and a higher occurrence of obesity [8].

Normal functioning of the thyroid gland is essential for successful conception and pregnancy. The impact of subclinical hypothyroidism during pregnancy is unclear. Some observational studies have shown adverse outcomes [9]. In a large prospective study, pregnant women with SCH were at a higher risk for placental abruption and preterm delivery compared with euthyroid women and also, their offspring were more likely to be admitted in the neonatal intensive care unit and have respiratory distress syndrome [10].

Overt hypothyroidism during pregnancy is associated with different adverse outcomes, including miscarriage, pre-eclampsia, placental abruption, preterm birth, low birth weight and reduced IQ in offspring [11-13].

Hypothyroidism is a chronic disease.  Chronic diseases have long and variable preclinical phases. The preclinical phase is that portion of the disease’s natural history during which the disease is potentially detectable, but unrecognized.

Lead time is the interval between the time of disease detection through screening and the time of disease recognition in the absence of screening.  The lead time produced by a screening program for a given individual depends on the time of screening, in relation to the preclinical phase, and on the sensitivity function of the screening test. Screening is the application of a test to detect condition in an individual who has no known signs or symptoms of that disease [14-15].

Although, most serum thyroid markers are straightforward to interpret and confirm the clinical impression of thyroid disorders (euthyroidism, hypothyroidism or hyperthyroidism), however, in an important subgroup of patients the results of serum thyroid markers can seem unclear, either by virtue of being discordant with the clinical picture or because they appear incongruent with each other [e.g. raised thyroid hormones (TH), but with non-suppressed thyrotropin (TSH); elevated TSH, but with normal TH] [16].

The most sensitive test for thyroid dysfunction is the serum TSH. Serum TSH measurements may yield misleading results for individuals with changing levels of thyroid hormones.

Non-thyroidal disorder, in field of acute and chronic inflammation process, can modify hypothalamic/pituitary function and thyroid hormone metabolism and consequently lead to thyroid test abnormalities, including both decreased and increased serum TSH levels. Among individuals with serious, acute illness, the serum TSH is less specific for thyroid disease because a serious illness alone can depress TSH secretion. The prevalence of both low and high serum TSH levels (with normal serum free T4 results) is increased in elderly subjects compared with younger people [17-20].  

TSH exhibits diurnal variation, with the lowest value in the late afternoon and highest value between midnight and 4 AM. Therefore, variations of serum TSH values within the normal range of up to 50% do not necessarily reflect a change in thyroid status [21].

There is now evidence linking cells of the immune system to the regulation of thyroid hormone activity in normal physiological conditions as well as during times of immunological stress. It is known that thyroid stimulating hormone (TSH) can be produced by many types of extra-pituitary cells — including T cells, B cells, splenic dendritic cells, bone marrow hematopoietic cells, intestinal epithelial cells and lymphocytes [22].

Studies showed that estrogen therapy/hormone therapy raises the circulating levels of thyroxine-binding globulin (TBG), thereby increasing the bound fraction and decreasing the free fraction of circulating thyroxine (T4) [23-24].  

Recently, it was reported that thyroid hormone receptors (TRs) are also present in human ovarian surface epithelium and act on ovarian follicles and show some slight localization in granulosa cells of ovarian follicles) [25].

Hypothyroidism (non-autoimmune) is commonly observed in chronic kidney disease (CKD) patients. CKD patients have low T3 and normal or reduced T4 levels, and consequently elevated TSH and attendant increase in thyroid gland volume [26-28].

Drugs can lead to thyroid dysfunction and abnormality in serum level of thyroid hormones with different mechanisms. Drugs may cause impairment in thyroxine absorption, may increase hepatic metabolism and cause hypothyroidism (carbamazepine, hydantoins, phenobarbital, rifampin); that may cause a decrease in thyroid hormone secretion and therefore, cause a higher dose requirement of thyroxine (aminoglutethimide, amiodarone, iodide-including and iodine-containing radiographic contrast agents, lithium, methimazole, propylthiouracil, sulfonamides, tolbutamide); or may decrease T4 5′-deiodinase activity (amiodarone, beta-adrenergic antagonists, glucocorticoids, propylthiouracil); they may increase or  decrease serum  thyroxine-binding globulin (TBG) concentration (estrogen, clofibrate, heroin/methadone, mitotane, tamoxifen,androgens/anabolic steroids, glucocorticoids, nicotinic acid, asparginase); and may cause protein-binding site displacement (furosemide, heparin, hydantoins, nonsteroidal anti-inflammatory drugs: fenamates, phenylbutazone, salicylates) [29-32].  

Thyroid ultrasonography (US) has established itself as a common and valuable tool in the evaluation and management of thyroid disorders. Ultrasound scanning is non-invasive, widely available, less expensive, and does not use any ionizing radiation [33]. Several studies demonstrated an association between hypoechogenicity at thyroid US and higher levels of serum TSH even in subjects without overt thyroid disease, suggesting decreased echogenicity as an early sign of thyroid dysfunction [34-36].

Limitations of sonography are the following:

The major limitation of ultrasound in thyroid imaging is that it cannot determine thyroid function, i.e., whether the thyroid gland is hypo-, hyper- or normal in function [33]. US is an observer-dependent method. An important source of error in ultrasonography depends on the technical skill of the operator. A diagnostic procedure performed by a poorly trained ultrasound technician or a well-trained technician making an error can produce inconsistent and incorrect results. Emergency US is particularly susceptible to errors, more than any other diagnostic imaging technique; in fact, the misinterpretation of sonographic images should be considered as a serious risk in US-based diagnosis [37].  

Technical limits of thyroid US are represented by thyroid nodules spreading into substernal, retroclavicular, intrathoracic or retrotracheal locations that may not be easily imaged with US. Sonography can be effective in detection of retrosternal goiter when it is in the upper mediastinum, however, location of the goiter below the bifurcation of the trachea limits the possibility of US [38].  

 Nowadays, the use of US scanners may detect fluid lesions 1 mm or more and solid lesion 2 to 3 mm or more in size in thyroid [39].

Another disadvantage of ultrasound technology involves body size of the patient. Sonography visualization problems can arise when the target area is deep within the body. The presence of gas also affects the visual quality of ultrasound images, as gas induces poor quality image output [40].  When performing thyroid US, proper compression is essential: insufficient force may lead to imaging artifacts. Failure to apply adequate pressure results in deeper lesions appearing more indistinct because of ultrasound attenuation.

Sonography procedure is generally safe, but may lead to the non-thermal, mechanical and thermal effect at the cellular and molecular levels.

Non-thermal effects of ultrasound include acoustic cavitation, by acceleration and movement of particles in liquid and the sudden release of energy, which can be sufficiently intense to disrupt molecular bonds. In molecular level, according to the frequency resonance hypothesis, ultrasound’s non-thermal action affects enzyme activity and possibly gene regulation in tissues through two different mechanisms. First, absorption of mechanical energy by a protein may produce a transient conformational shift in the 3-dimensional structure and alter the protein’s functional activity. Second, the resonance properties of the wave may dissociate a multi-molecular complex, thereby disrupting the complex’s function [41].

Human body can be considered as an open, non-isolated superorganism. Thyroid diseases are multifactorial and complex disorders.  As with other chronic diseases, there is a multifactorial etiology with a complex interaction of environmental and physiological factors in development of thyroid disease [42-43].   

Probably, any physiological and pathological phenomena basically cannot be explained on separated factors and mechanism alone, without the consideration of other aspect of life. Reduction of life phenomena to separated biological levels of organization of living organisms means almost totally refusing consideration of such typical features living systems as nonlinearity, networking, stochasticity, emergence and others aspects of complex systems [44]. In the recent decade the use of informative-wave technologies has been widely applied into practical medicine. The 3D-Nonlinear Bio Resonance Feed-back Diagnostics Systems (3D NLS BR) have been extensively used recently and are gaining ever growing popularity. Non-linear (NLS) diagnostic approaches are based on a new physics of quantum-entropic interactions or Quantum-entropic logic theory, that was previously described [43,45-46].  

To our knowledge, to date, no studies have been published regarding using 3D-NLS BR Systems in screening of thyroid glands. 

The purpose of this study was to test individuals with no dominant clinical symptoms with spectral-entropy analysis (SEA) parameters using 3D-NLS BR diagnostic method. This study was carried out in the medical scientific center of bioelectric and bio-feedback technologies in cooperation with Institute of Practical Psychophysics (IPP). 

Participants and Methods

This experimental study was evaluated and approved by our Institutional Review Board, and was carried out from 2015 till 2017. The study was conducted on 1,200 adults (aged from 18 to 60 years old), who annually underwent thyroid hormone measurement with normal results and did not have any history of thyroid disease or other systemic diseases in their past medical history. 

Inclusion and Exclusion Criteria:

In this study, our team initially used biophysical markers for evaluation of thyroid dysfunction. For confirmation or exclusion of an underlying thyroid disorder, the NLS-3D BR system was used for evaluation 3 biophysical spectral-entropy parameters:

1) Fleindler’s 12-point polychrome scale (FI).,2) Entropy level (EI)., and 3) Noise/information index(NII).

The NLS-bioresonance diagnostic is carried out using the hardware-software complex metatron device registration no FSNO 022a2005/222105 using the Spectral Entropy Analysis (SEA) parameters, which were described in the previous article [43].

Individuals who showed FI- from 1 to 3, EI- from 1-4 and NII- less than 1, were classified as the Control Group (with this range indexes, it was expected the thyroid function and structure to be normal. Individuals, who showed FI: 4 or more than 4., EI: 5 and more than 5, and NII equal 1 or more than 1, were classified as the Case Group (with this range indexes, it was expected the thyroid function and structure to be abnormal (see Table1).

Table1:

Groups	Fleindler’s index(FI)	Entropy index(EI)	Noise/information index(NII)
Control=normal	3	Up to 4	Less than 1
Case	4 and more	5 and more	Equal 1 and more

After that, according to greatness of biophysical parameters, the Case Group was subdivided into two subgroups, namely A and B. All cases with the FI=4, EI=5 and NII=1 were assigned to Case Group A, and those with FI=5 or more than 5., EI=6-7 and NII-more than 1 were assigned to case group B (cases B, were expected to show worse thyroid structural and functional abnormality than Cases A) (see Table2).

Table2:

Cases	Fleindler’s index(FI)	Entropy index	Noise/information index(NII)
Case A	4	5	Equal 1
Case B	5 or more	6 -7	more than1

According to the inclusion and exclusion criteria, after scanning with 3D NLS-BR system, participants were subdivided into two groups namely case group and control group. Case group consisted of 400 individuals, 300 of them were women (75%) and 100 of them were men (25%) (average age 42±28 years). Control group consisted of 800 individuals. 562 of them were women (70.25%) and 238 of them were men (29.85%). (average age 43±34 years). All participants were from Tehran city, Iran.

Ultra-sound screening of thyroid gland:

After evaluation with NLS-bioresonance diagnostic method, the blood sampling was sent to reference lab for TSH, T3, T4 and anti-TPO testing. In addition, ultrasound examinations were performed on both Control and Case groups. Thyroid ultrasound (US) was performed using the HDI 5000 or IU 22 (Philips medical system, Best, the Netherlands) with a 7.5 MHz linear probe. A reference radiologic center with 15 years of experience in thyroid US interpreted the US results in consensus. The Case Group and Control Group were classified into three groups based on the US results: group A (patients who had normal US) and group B (patients who were hypoechoic relative to the submandibular gland, but hyperechoic relative to strap muscle), and finally group C (It defined when the US showed multiple scattered small hypoechoic nodules in the whole thyroid parenchyma).

Decreased echogenicity was defined when more than two-thirds of the whole thyroid parenchyma showed lower echogenicity than that of the individual patient’s submandibular gland by visual assessment. When the echogenicity of submandibular gland was abnormal due to sialoadenitis or connective tissue disease, the echogenicity of parotid gland was considered as standard.

Laboratory procedures

Thyroid status was evaluated from blood samples by laboratory serum markers. Laboratory tests for thyroid function (serum levels of T3, T4, and TSH) and serum thyroid anti-thyroid peroxidase autoantibody [anti-TPO) were evaluated in these subjects.

Total T3 and T4 and TSH were determined by standard Electro-chemiluminescence (ECL) technique, supplied by Architect-Abott. Reference ranges(micg/dl) for total thyroxine (T4) were as follows: In 1-3 days (11.8-22.6), 4-14 day (9.8-16.6)., 15 day-4 months (7.2-14.4)., 5-12 month (7.8-16.5)., 13 month-5 years (7.3-15)., 6-10 year (6.4-13.3 and more than 11 years (5.1-14.1).

Reference ranges (ng/ml) for total thyroxine (T3) were as follows: In 1-3 days (1-7.4), 4 day- 5 years (1.05-2.69)., 6-10year (0.94-2.41)., 11-15year (0.83-2.13)., and more than 16 years (0.58-1.59).

For TSH value: between 4.7 and 10 mU/L is considered subclinical hypothyroidism., over 10 mU/L is overt (symptomatic) hypothyroidism., between 1.5 and 2.0 mU/L is suggestive of thyroid dysfunction., between 0.1 and 0.5 mU/L is considered subclinical hyperthyroidism and less than 0.1 mU/L is overt hyperthyroidism.

Anti-TPO (IU/ml) was determined by Elisa technique, supplied by Tecan-sunrise: less than 50 IU/ml –negative., 50-75 borderline and more 75 considered as positive.

Statistical analysis

Data analyses were conducted in SPSS (version 25) using Mann-Whitney U test (this nonparametric test was employed in this study, because the data were classified as nominal scales) to confirm whether the differences occurred between groups.

Results: Table 3 shows the biochemical and ultra-sound outcome in the case group and the control group.  

In the Control Group, all 800 (100%) individuals had normal levels of TSH and T4 hormones, and 784 individuals (98%) had normal and 16(2%) showed the positive level of anti-TPO.

In the Case Group, out of 400 cases (A+B), 362 (90%) had normal levels of TSH and 38(10%) cases had borderline levels of TSH. 396 (99%) had normal levels of T4 and 4(1%) had the abnormal levels of T4. For Anti-TPO marker, 230(57.5%) at the normal (42.5%levels, normal and abnormal level, respectively (see Table 3).

Table3:

Groups	group		Biochemical						Ultrasono
			TSH micIU/ml		T4 micg/dl		Anti TPO IU/ml		Hypo/echo		nodule
			0.4-4.6	4.7-10	5-14	≥14.2	≤50	≥51	yes	NUS	yes	no
Case	A+ B= 400	N	362	38	396	4	230	170	389	11	116	284
		%	90	10	99	1	57.5	42.5	97	3	29	71
Control	C= 800	N	800	0	800	0	784	16	800	0	800	0
		%	100	0%	100	0%	98%	2%	0%	100%	0%	100

Serum level of T3 hormone in all participants was normal. Sonography of 800 participants in the Control group showed 100% had normal pattern, compared to the Case group, in which, 394 (97%) had decreased echogenicity and 116 (29%) had nodular pattern of thyroid gland.

Mann Whitney U test showed a difference between Case group and Control group in biochemical and sonography at the significance level of .05. (Table 4).

 

Table 4. Descriptive statistics of biochemical parameters
Case Group (n=400)				Control Group (n=800)
Min	Max	M	SD	Min	Max	M	SD
1.00	2.00	1.10	0.29	1.00	1.00	1.00	0.00
1.00	2.00	1.91	0.29	2.00	2.00	2.00	0.00
1.00	2.00	1.01	0.10	1.00	1.00	1.00	0.00
1.00	2.00	1.99	0.10	2.00	2.00	2.00	0.00
1.00	2.00	1.43	0.49	1.00	2.00	1.02	0.14
1.00	2.00	1.58	0.49	1.00	2.00	1.98	0.14
1.00	2.00	1.97	0.16	1.00	2.00	1.00	0.04
1.00	2.00	1.03	0.16	2.00	2.00	2.00	0.00
1.00	2.00	1.71	0.45	2.00	2.00	2.00	0.00
Note: Mann Whitney U test showed a difference between group case group and control group in biochemical and sonography at the significance level of .05

On the other hand, according to Table 2, when all cases were divided into two groups, the following data collected of the 400 cases (A+B), 280 and 120 were classified as A and B, respectively.

In the A cases, 280 (100%) had normal levels of TSH and T4 thyroid hormone.  For Anti-TPO marker, 181 (64%) had negative, 32 (12%) were borderline, and 67(24%) had positive serum levels.

In the B cases, 82 (68%) had normal, 38 (32%) had borderline levels of TSH serum, 116 (97%) had normal, and 4 (3%) had abnormal T4 serum levels.  For Anti-TPO marker, 49 (41%) had negative, 0% were borderline, and 71 (59%) had positive serum levels. In the Control group, 800 (100%) individuals had normal levels of TSH and T4 hormones, 784 persons (98%) had normal levels, and 16(2%) had positive levels of anti-TPO (see Table5).

In sonography, in Case A group, 269 (96%) had decreased echogenicity,11 (4%) had normal echo thyroid pattern, and 59 (21%) with and 221 (79%) without nodular pattern of thyroid gland, compared with Case B group, in which, 120 (100%) of cases had decreased echogenicity and 57(48%) showed nodular pattern of thyroid gland. Of the 800 controls, 100% had normal pattern of thyroid gland (Table 5).

A Mann-Whitney U test was run to determine if there were significant differences between case group and control group in biochemical markers. As can be seen in Table 4, there were significant differences in all biochemical markers between case group and control group.

Table5:

 

Groups	group		biochemical							Ultrasono
			TSH micIU/ml		T4 micg/dl		Anti TPO IU/ml			Hypo/echo		Nodule
			0.4-4.6	4.7-10	5-14	≥14.2	Ng	Bl	Po	yes	NUS	yes	no
Case	A=280	N	280	0	280	0	181	32	67	269	11	59	221
		%	100	0	100%	0%	64%	12%	24%	96%	4%	21%	79%
	B=120	N	82	38	116	4	49	0	71	120	0	57	63
		%	68%	32%	97%	3%	41%	100	59	100%	0%	48%	52%
Control	C=800	N	800	0	800	0	784	7	9	800	0	800	0
		%	100	0%	100%	0%	98%	0.8%	1.2%	0%	100%	0%	100

Mann Whitney U test showed a difference between group A and group B in biochemical and sonography at the significance level of .05 (Table6).

 

Table 6. Mean and standard deviation values of  biochemical and sonography for two sub-scales of A and B groups.
Biophysical Index			TSH1	TSH2	T43	T44	AntiTPO≤50	AntiTPO ≥50	NUS5	HHPEP6	Nodule
Fleindler’s index	Case A (n=280)	Mean	1.00	2.00	1.00	2.00	1.35	1.65	1.96	1.04	1.79
		SD	0.00	0.00	0.00	0.00	0.48	0.48	0.19	0.19	0.41
	Case B (n=120)	Mean	1.32	1.68	1.03	1.97	1.59	1.41	2.00	1.00	1.53
		SD	0.47	0.47	0.18	0.18	0.49	0.49	0.00	0.00	0.50
Entropy index	Case A (n=280)	Mean	1.00	2.00	1.00	2.00	1.35	1.65	1.96	1.04	1.79
		SD	0.00	0.00	0.00	0.00	0.48	0.48	0.19	0.19	0.41
	Case B (n=120)	Mean	1.32	1.68	1.03	1.97	1.59	1.41	2.00	1.00	1.53
		SD	0.47	0.47	0.18	0.18	0.49	0.49	0.00	0.00	0.50
Noise/ information index	Case A (n=253)	Mean	1.00	2.00	1.00	2.00	1.29	1.71	1.96	1.04	1.87
		SD	0.00	0.00	0.00	0.00	0.45	0.45	0.20	0.20	0.34
	Case B (n=147)	Mean	1.26	1.74	1.03	1.97	1.66	1.34	2.00	1.00	1.44
		SD	0.44	0.44	0.16	0.16	0.48	0.48	0.00	0.00	0.50
Note: Mann Whitney U test showed a difference between group A and group B in biochemical and sonography at the significance level of .05
1= TSH (normal range: 0.4-4.6 micIU/ml), 2=(more than 4.7 to 10 -subclinical hypothyroidism), 3= T4 micg/dl (5-14 normal range), 4=(more than 14.2 abnormal range),  5= normal sonography, 6= Heterogeneous and Hypoechoic Parenchymal Echo Pattern

A Mann-Whitney U test was run to determine if there were significant differences between group A and group B in biochemical markers and sonography. As shown in Table 5, there were significant differences in all biochemical markers and sonography between group A and group B.

Discussion:

This experimental study showed the evidence for a useful application of 3D-NLS BR system in screening of thyroid dysfunction. To the best of the authors’ knowledge, this is the first study on this topic. Subclinical hypothyroidism (SH), also known as isolated hyperthyrotropinemia, is often associated with adverse health risk, such as cardiovascular disorder, high cholesterol, hypertension, endothelial dysfunction and atherosclerotic. The ischemic abnormalities are probably related to long-term consequences of a slowly progressing development of hypothyroidism [47- 49].

The most useful marker for the assessment of thyroid hormone action is TSH level.  It is defined as an elevated level of TSH in the framework of free T4 and T3 levels within the normal reference range. But the secretion and action of TSH are affected by many factors such as non-thyroidal disorder, immune system disturbance, infections, different medications, aging etc.

Meanwhile, the key role of the clinical laboratory is to report accurate results for diagnosis of disease. This goal has, to a great extent been achieved for routine biochemical tests, but not for immunoassays. Immunoassays may lack adequate specificity and accuracy and remained susceptible to interference from endogenous immunoglobulin antibodies, causing false, and clinically misleading results [50- 52]. Interference in immunoassay is one factor that contributes to the uncertainty of medical testing. In recent years, it has become evident that a consensus on the exact limits for cut-off between normal and subclinically hypothyroid individuals is not currently possible. Reference populations used as the basis for a normal range are highly different in relations of genetics and epigenetics background, and environmental factors such as iodine intake, age, gender and presence of thyroid autoantibodies and should not be confused with cut-off limits [53].

Then, although, the TSH level is a useful marker for the assessment of peripheral thyroid hormone action, but the values should be interpreted carefully, and TSH level alone could not be used to assess the thyroid hormone action in vivo.

Thyroid ultrasonography (US) also plays a key role in the diagnosis of thyroid-related diseases. For appropriate thyroid examination, a high frequency (10-15 MHz) linear transducer is recommended [54-55]. Ultrasound may have biological thermal and non-thermal effects in body tissues. All waves carry energy and momentum. Acoustic radiation force, radiation torque, acoustic streaming, shock waves and cavitation are considered non-thermal effects of the ultrasound.  Acoustic radiation force is a physical phenomenon resulting from the interaction of an acoustic wave with an obstacle placed along its path. Cavitation is described as the formation and oscillation of a gas bubble and is recognized as a major cause of ultrasound-induced mechanical and thermal effects.  Tissues naturally containing gas bodies, such as the lung and intestine, are more sensitive to the bio-effects of ultrasound exposure because of the presence of gas. Cavitation-related bio-effects are more dependent on frequency [56].

Several animal studies observed hemorrhage in lung tissues after ultrasound exposure (at 1.2 MHz, 3-min exposure) in mice, rabbit and pigs [57-59]. Acoustic cavitation can be generated in a wide variety of intestinal environments, as this part of body contain gas bodies located in a fluid-like medium [60].

In in-vivo studies with laboratory animals revealed that the destruction of blood cells in the presence of ultrasound contrast agents at diagnostic levels of ultrasound has occurred [61]. Fetal red blood cells are more susceptible to lysis from exposure to ultrasound in the presence of contrast agents in vitro [62].  

Cells and tissue can be damaged when exposed to ultrasound pulses if they lie close to a region of gas already contained within tissue. The shear forces generated at the tissue/gas interface may be sufficient to cause damage. But, it is generally accepted that diagnostic ultrasound does not have serious side effects in organisms, but as mentioned before, have some limitation.

Compared with ultrasound, 3D NLS systems do not have such limitations as observer-dependent and have a possibility for evaluation of functional status and structure of thyroid gland, simultaneously.

Analysis data extracted by using 3D NLS bioresonance feed-back system showed that Spectral Entropy Analysis (SEA) parameters are valuable markers for early detection of thyroid–related disorders and can be used in screening of asymptomatic patients.

In general, screening has two major objectives. One is the early detection of the disease —before the development of symptoms—at a point when treatment is more effective and less expensive. And the second objective in screening is to identify risk factors that render an individual at a higher risk for developing a disease, with the goal of modifying the risk factors to prevent or reduce the disease [63-66]. Neither the TSH serum level, nor the ultrasound method can meet both criteria in screening, nevertheless the 3D NLS BR system method, because of its holistic characterization can overcome both of these criteria.

Conclusion:

 According to the findings obtained from this study, 3D NLS bioresonance feed-back system is clinically effective in screening of hypothyroidism.

Acknowledgements:

This study was supported by grants from The Medical Scientific Center for Development of Bioelectric and Bio-feedback Technologies, Tehran, Iran

Conflict of interest: Authors have no conflict of interest.

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