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Women's Health Science Journal Research Article 22 min read

Possible Role of Inadequate Quantities of Intra-Thyroidal Cobalt, Rubidium and Zinc in the Etiology of Female Subclinical Hypothyroidism

Vladimir Zaichick1* and Sofia Zaichick 1*, 2
* Corresponding author
ISSN: 2639-2526  10.23880/whsj-16000108  Received: January 20, 2018  Published: February 21, 2018
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Keywords
Subclinical hypothyroidism Female Thyroid Trace Elements Neutron activation analysis
Abstract

Background: Subclinical hypothyroidism does affect fertility. The prevalence of subclinical hypothyroidism is 10-15 times more common in women than in men. Trace elements play important roles in thyroid function and fertility. Objective: The aim of this exploratory study was to evaluate whether significant differences of trace element contents exists between female and male thyroids and how they can be related to the etiology of subclinical hypothyroidism. Methods: Thyroid tissue levels of ten trace elements: silver, cobalt, chromium, iron, mercury, rubidium, antimony, scandium, selenium, and zinc were prospectively evaluated in 105 healthy persons (33 females and 72 males). Measurements were performed using instrumental neutron activation analysis with high resolution spectrometry of long-lived radionuclides. Tissue samples were divided into two portions. One was used for morphological study while the other was intended for trace element analysis. Results: It was found that content of cobaltin thyroid of females was significantly higher than that of males, while contents of rubidiumand zinc were lower. Conclusions: Inappropriate content of intra-thyroidal cobalt, rubidiumand zinc can be associated with the etiology of female subclinical hypothyroidism.

Vladimir Zaichick1* and Sofia Zaichick 1,2

Russia

Northwestern University, USA

60289; Fax: +7 (495) 956 1440; E-mail: vezai@obninsk.com hypothyroidism.

the other was intended for trace element analysis.

contents of rubidiumand zinc were lower.

female subclinical hypothyroidism.

Lived Radionuclides Fluorescent Analysis; SRM: Standard Reference Material; CRM: Certified Reference Material; BSS: Biological Synthetic Standards.

Introduction

Adequate thyroid function is important to maintain normal reproduction, because thyroid dysfunction affects fertility in various ways resulting in abnormal ovulatory cycles, luteal phase defects, high prolactin levels, and sex hormone imbalances [1, 2]. Therefore, normal thyroid function is necessary for fertility, and to sustain a healthy pregnancy [2]. From large population studies, which measured thyroid function, and systematic reviews of this subject carried out in the 1990s to 2010s, it is known that untreated hypothyroidism is a common condition all over the world [2, 3, 4, 5, 6, 7, 8, 9, 10]. The prevalence of subclinical hypothyroidism (SCH) is between 1% and 10% in different countries [2, 3, 4, 5, 6, 7, 8, 9, 10] and almost everywhere it is 10-15 times more common in women than in men [2, 4, 9]. Form such a great gender-related difference in the prevalence of SCH arises a question about a specific sensitivity of female thyroid tissue to some external and internal factors. Although the etiology of SCH and other thyroidal disorders is unknown in detail, several risk factors including deficiency or excess of such micronutrients as iodine (I) has been well identified [11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22]. Besides I involved in thyroid function, other trace elements (TE) also play important roles such as stabilizers, structural elements, maintenance and regulation of cell function, gene regulation, enzyme cofactors, activation or inhibition of enzymatic reactions, normal peripheral utilization of thyroid hormones and regulation of cell membrane function [23]. Essential or toxic properties of TE depend on tissue-specific need or tolerance, respectively [24]. Both TE deficiencies as well as overexposures may disturb the thyroidal cell functions [24]. The reliable data on TE mass fractions in normal human thyroid separately for female and male gland is apparently extremely limited. There are a few studies regarding TE content in human thyroid, using chemical techniques and instrumental methods [25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35]. However, the majority of these data are based on measurements of processed tissue and in many studies tissue samples are ashed before analysis. In other cases, thyroid samples are treated with solvents (distilled water, ethanol etc) and then are dried at a high temperature for many hours. There is evidence that certain quantities of TE are lost as a result of such treatment [36, 37, 38]. Moreover, only a few of these studies employed quality control using certified/standard reference materials (CRM/SRM) for determination of the TE mass fractions. Sample-nondestructive technique such as instrumental neutron activation analysis with high resolution spectrometry of long-lived radionuclides (INAA-LLR) is good alternatives for multi- element determination in the samples of thyroid parenchyma. This work had three aims. The primary purpose of this study was to determine reliable values for such TE as silver (Ag), cobalt (Co), chromium (Cr), iron (Fe), mercury (Hg), rubidium (Rb), antimony (Sb), scandium (Sc), selenium (Se), and zinc (Zn) contents in intact (normal) thyroid gland of apparently healthy persons using INAA-LLR analysis. The second aim was to compare the levels of TE in the thyroid tissue of all females and males investigated in the study. The final aim was to compare the levels of TE in the thyroid tissue of females and males in age group 1 (≤40 years) and in age group 2 (>40 years). All studies were approved by the Ethical Committees of the Medical Radiological Research Centre, Obninsk.

Material and Methods

Samples of the human thyroid were obtained from randomly selected autopsy specimens of 33 females (European-Caucasian, aged 3.5 to 87 years) and 72 males (European-Caucasian, aged 2.0 to 80 years). All the deceased were citizens of Obninsk and had undergone routine autopsy at the Forensic Medicine Department of City Hospital, Obninsk. Age ranges for subjects were divided into two age groups, with group 1 (≤40 years), and group 2 (>40 years). For females in group 1 (n=11) mean age (±standard error of mean, SEM) was 30.9±3.1 years and in group 2 (n=22) mean age was 66.3±2.7 years. For males in group 1 (n=36) mean age was 22.5±1.4 years and in group 2 (n=36) mean age was 52.4±2.4 years. These groups were selected to reflect the condition of thyroid tissue in the children, teenagers, young adults and first period of adult life (group 1) and in the second period of adult life as well as in old age (group 2). The available clinical data were reviewed for each subject. None of the subjects had a history of an intersex condition, endocrine disorder, or other chronic disease that could affect the normal development of the thyroid. None of the subjects were receiving medications or used any supplements known to affect thyroid trace element contents. The typical causes of sudden death of most of these subjects included trauma or suicide and also acute untreated illness (cardiac insufficiency, stroke, embolism of pulmonary artery, alcohol poisoning). All right lobes of thyroid glands were divided into two portions using a titanium scalpel [39]. One tissue portion was reviewed by an anatomical pathologist while the other was used for the ChE content determination. A histological examination was used to control the age norm conformity as well as the unavailability of microadenomatosis and latent cancer.

After the samples intended for TE analysis were weighed, they were freeze-dried and homogenized [40, 41, 42]. The pounded sample weighing about50 mg was used for trace element measurement by INAA-LLR. The samples for INAA-LLR were wrapped separately in a high-purity aluminum foil washed with rectified alcohol beforehand and placed in a nitric acid-washed quartz ampoule. To determine contents of the TE by comparison with a known standard, biological synthetic standards (BSS) prepared from phenol-formaldehyde resins were used [43]. In addition to BSS, aliquots of commercial, chemically pure compounds were also used as standards. Ten certified reference material IAEA H-4 (animal muscle) and IAEA HH-1 (human hair) sub- samples weighing about 50 mg were treated and analyzed in the same conditions that thyroid samples to estimate the precision and accuracy of results. A vertical channel of nuclear reactor was applied to determine the content of Ag, Co, Cr, Fe, Hg, Rb, Sb, Sc, Se, and Zn by INAA-LLR. The quartz ampoule with thyroid samples, standards, and certified reference material was soldered, positioned in a transport aluminum container and exposed to a 24-hour neutron irradiation in a vertical channel with a neutron flux of 1.3⋅1013 n⋅cm-2⋅s-1. Ten days after irradiation samples were reweighed and repacked. The samples were measured for period from 10 to 30 days after irradiation. The duration of measurements was from 20 min to 10 hours subject to pulse counting rate. The gamma spectrometer included the 100 cm3 Ge (Li) detector and on-line computer-based MCA system. The spectrometer provided a resolution of 1.9 keV on the 60Co 1332 keV line. Details of used nuclear reactions, radionuclides, and gamma-energies were presented in our earlier publications concerning the INAA chemical element contents in human prostate and scalp hair [44, 45, 46, 47, 48]. A dedicated computer program for INAA mode optimization was used [49]. All thyroid samples were prepared in duplicate, and mean values of TE contents were used in final calculation. Using Microsoft Office Excel software, a summary of the statistics, including, arithmetic mean, standard deviation, standard error of mean, minimum and maximum values, median, percentiles with 0.025 and 0.975 levels was calculated for TE contents in thyroid tissue samples of females and males. The difference in the results between females and males (age group 1 and 2 combined), as well as between females and males separately in age group 1 and group 2 was evaluated by the parametric Student’s t-test and non-parametric Wilcoxon-Mann-Whitney U- test.

Results

(Table 1) depicts our data for Ag, Co, Cr, Fe, Hg, Rb, Sb, Sc, Se, and Zn mass fractions in ten sub-samples of IAEA H-4 (animal muscle) and IAEA HH-1 (human hair) certified reference material and the certified values of this material.

IAEA H-4 animal muscleThis work resultsIAEA HH-1 human hairThis work results
Element
95% confidence intervalM±SD95% confidence intervalM±SD
Ag-0.033±0.0080.19b0.18±0.05
Co0.0027b0.0034±0.00085.97±0.42a5.4±1.1
Cr0.06b0.071±0.0100.27b≤0.3
Fe49.1±6.5a47.0±1.023.7±3.1a25.1±4.3
Hg0.014b0.015±0.0041.70±0.09a1.54±0.14
Rb18.7±3.5a23.7±3.70.94b0.89±0.17
Sb0.0056b0.0061±0.00210.031b0.033±0.009
Sc0.0059b0.0015±0.0009--
Se0.28±0.08a0.281±0.0140.35±0.02a0.37±0.08
Zn86.3±11.5a91±2174±9a173±17

Table 1: Neutron activation analysis data of trace element contents in certified reference material IAEA H-4 (animal muscle) and

Table 1: Neutron activation analysis data of trace element contents in certified reference material IAEA H-4 (animal muscle) and IAEA HH-1 (human hair) compared to certified values ((mg/kg, dry mass basis). M – arithmetical mean, SD – standard deviation, a – certified values, b – information values. (Table 2) presents certain statistical parameters (arithmetic mean, standard deviation, standard error of mean, minimal and maximal values, median, percentiles with 0.025 and 0.975 levels) of the Ag, Co, Cr, Fe, Hg, Rb, Sb, Sc, Se, and Znmass fractionin normal thyroid tissue of female and male.

GenderElementMSDSEMMinMaxMedianP 0.025P 0.975
Males
n=72		Ag0.01560.01550.00210.00170.080.01040.00180.0661
Co0.03520.02340.00310.00460.1240.03020.01130.101
Cr0.520.2860.0410.131.30.4140.1520.98
Fe22296125148722176.1432
Hg0.04610.03910.00530.00910.180.03240.01020.15
Rb7.894.560.582.2429.46.862.7318.2
Sb0.1080.0760.010.00470.3080.09650.00950.291
Sc0.00510.00360.00120.00050.01180.00440.00070.0112
Se2.361.340.170.535.81.960.8045.7
Zn103435.53422194.640.5200
Females
n=33		Ag0.0140.00930.0020.00120.03310.0130.00210.0321
Co0.05050.03220.00640.0170.140.04050.01830.13
Cr0.5730.2460.0490.291.220.4880.3031.11
Fe232112226351219964.8480
Hg0.03290.02460.00510.00650.10.02630.00790.1
Rb6.162.420.481.1112.86.32.3810.8
Sb0.1160.0630.0120.01150.2480.1080.01830.247
Sc0.00420.0040.00120.00020.01430.00320.00030.0124
Se2.221.190.230.4395.322.070.7734.85
Zn85.7387.448.11668322.9156

Table 2: Some statistical parameters of Ag, Co, Cr, Fe,Hg, Rb, Sb, Sc,Se, and Znmass fractions (mg/kg, dry mass basis) in normal

Table 2: Some statistical parameters of Ag, Co, Cr, Fe,Hg, Rb, Sb, Sc,Se, and Znmass fractions (mg/kg, dry mass basis) in normal thyroid tissue of females and males. M – arithmetic mean, SD – standard deviation, SEM – standard error of mean, Min – minimum value, Max – maximum value, P 0.025 – percentile with 0.025 level, P 0.975 – percentile with 0.975 level. The comparison of our results with published data for Ag, Co, Cr, Fe, Hg, Rb, Sb, Sc, Se, and Znmass fraction in normal human thyroid is shown in Table 3.

This workPublished data [Reference]
ElementMales and femalesMedian of meansMinimum of means M orMaximum of means M or
M±SD(n)*M±SD, (n)**M±SD, (n)**
Ag0.015±0.0140.25 (12)0.000784 (16) [26]1.20±1.24(105) [27]
Co0.040±0.0270.34 (17)0.026±0.031 (46) [28]70.4±40.8 (14) [29]
Cr0.54±0.270.69 (17)0.105 (18) [30]24.8±2.4 (4) [31]
Fe225±100252 (21)56 (120) [32]2444±700 (14) [29]
Hg0.042±0.0360.08(13)0.0008±0.0002(10) [33]396±40 (4) [31]
Rb7.4±4.112.3 (9)≤0.85 (29) [33]294±191 (14) [29]
Sb0.111±0.0720.105 (10)0.040±0.003(-) [34]4.0 (-) [35]
Sc0.0046±0.00380.009 (4)0.0018±0.0003(17) [36]0.0135±0.0045 (10) [33]
Se2.32±1.292.61 (17)0.95±0.08(29) [33]756±680 (14) [29]
Zn97.8±42.3118 (51)32(120) [32]820±204 (14) [29]

Table 3: Median, minimum and maximum value of means of Ag, Co, Cr, Fe, Hg, Rb, Sb, Sc, Se, and Zn contents in normal thyroid acco

Table 3: Median, minimum and maximum value of means of Ag, Co, Cr, Fe, Hg, Rb, Sb, Sc, Se, and Zn contents in normal thyroid according to data from the literature in comparison with our results (mg/kg, dry mass basis). M –arithmetic mean, SD – standard deviation, (n)* – number of all references, (n)** – number of samples. The ratios of means and the difference between mean values Ag, Co, Cr, Fe, Hg, Rb, Sb, Sc, Se, and Zn mass fractions in normal thyroid of females and males are presented in Table 4. Because, in our previous studies age-dependents of many TE in thyroid gland was found [50, 51, 52, 53], the comparison between TE contents in thyroid of females and males separately in age group 1 and in age group 2 was also performed (Tables 4-6).

Thyroid tissueRatio
Element
Males 2.0-80 years n=72Females 3.5-87 years n=33Student’s t-test pU-test pFemales to Males
Ag0.0156±0.00210.0140±0.00200.586>0.050.9
Co0.0352±0.00310.0505±0.00640.0390.011.43
Cr0.520±0.0410.573±0.0490.411>0.051.1
Fe222±12232±220.692>0.051.05
Hg0.0461±0.00530.0329±0.00510.079>0.050.71
Rb7.89±0.586.16±0.480.0240.010.78
Sb0.108±0.0100.116±0.0120.649>0.051.07
Sc0.0051±0.00120.0042±0.00120.59>0.050.82
Se2.36±0.172.22±0.230.633>0.050.94
Zn103±5.585.7±7.40.0690.050.83

Table 4: Differences between mean values (M±SEM) of Ag, Co, Cr, Fe,Hg, Rb, Sb, Sc,Se, and Znmass fraction (mg/kg, dry mass basis)

Thyroid tissueRatio
Element
Males (MG1) n=44Females (FG1) n=11Student’s t-test pU-test pFG1/MG1
Ag0.0160±0.00320.0143±0.00320.7>0.050.89
Co0.0374±0.00460.0328±0.00420.467>0.050.88
Cr0.502±0.0510.567±0.0650.441>0.051.13
Fe224±16172±220.0780.050.77
Hg0.0439±0.00710.0275±0.00460.058>0.050.63
Rb7.96±0.614.95±0.580.00110.010.62
Sb0.109±0.0120.0880±0.00960.174>0.050.81
Sc0.0052±0.00120.0026±0.00170.32>0.050.5
Se2.05±0.181.86±0.270.565>0.050.91
Zn102±6.659.8±8.70.00080.010.59

Table 5: Differences between mean values (M±SEM) of Ag, Co, Cr, Fe,Hg, Rb, Sb, Sc,Se, and Znmass fraction (mg/kg, dry mass basis)

Table 5: Differences between mean values (M±SEM) of Ag, Co, Cr, Fe,Hg, Rb, Sb, Sc,Se, and Znmass fraction (mg/kg, dry mass basis) in normal thyroid tissue of males and females aged ≤40 years. M – arithmetic mean, SEM – standard error of mean, t-test - Student’s t-test, U-test - Wilcoxon-Mann-Whitney U-test, Statistically significant values are in bold.

Thyroid tissueRatio
ElementMales (MG2)Females (FG2)Student’s t-testU-test
FG2/MG2
n=28n=22pp
Ag0.0148±0.00190.0138±0.00270.753>0.050.93
Co0.0317±0.00310.0644±0.00960.0050.012.03
Cr0.548±0.0700.578±0.0730.767>0.051.05
Fe218±19279±310.105>0.051.28
Hg0.0491±0.00820.0370±0.00840.311>0.050.75
Rb7.75±1.257.05±0.630.62>0.050.91
Sb0.107±0.0180.136±0.0190.276>0.051.27
Sc0.0049±0.00240.0045±0.00140.868>0.050.92
Se2.89±0.322.48±0.340.396>0.050.86
Zn104±10104.7±8.40.98>0.051.01

Table 6: Differences between mean values (M±SEM) of Ag, Co, Cr, Fe,Hg, Rb, Sb, Sc,Se, and Znmass fraction (mg/kg, dry mass basis)

Table 6: Differences between mean values (M±SEM) of Ag, Co, Cr, Fe,Hg, Rb, Sb, Sc,Se, and Znmass fraction (mg/kg, dry mass basis) in normal thyroid tissue of males and females aged >40 years. M – arithmetic mean, SEM – standard error of mean, t-test - Student’s t-test, U-test - Wilcoxon-Mann-Whitney U-test, Statistically significant values are in bold.

Discussion

Precision and Accuracy of Results

Good agreement of the Ag, Co, Cr, Fe, Hg, Rb, Sb, Sc, Se, and Zncontents analyzed by INAA-LLR with the certified data of CRM IAEA H-4 and IAEA HH-1 (Table 1) indicates an acceptable accuracy of the results obtained in the study of TE of the thyroid presented in Tables 2– 6. The mean values and all selected statistical parameters were calculated for tenTE (Ag, Co, Cr, Fe, Hg, Rb, Sb, Sc, Se, and Zn) mass fractions in thyroid of female and male (Table 2).

Comparison with Published Data

Values obtained for Cr, Fe, Hg, Rb, Sb, Sc, Se, and Zn contents in the normal human thyroid (Table 3) agree well with median of mean values reported by other researches [25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35]. The obtained means for Ag and Co were almost one order of magnitude lower median of previously reported means but inside the range of means (Table 3). Data cited in Table 3 also includes samples obtained from patients who died from different non-endocrine diseases. A number of values for TE mass fractions were not expressed on a dry mass basis by the authors of the cited references. However, we calculated these values using published data for water (75%) and ash (4.16% on dry mass basis) contents in thyroid of adults [27, 54]. The range of means of Ag, Co, Cr, Fe, Hg, Rb, Sb, Sc, Se, and Zn level reported in the literature for normal human thyroid vary widely (Table 3). This can be explained by a dependence of TE content on many factors, including the region of the thyroid, from which the sample was taken, age, gender, ethnicity, and mass of the gland. Not all these factors were strictly controlled in cited studies. Another and, in our opinion, leading cause of inter-observer variability can be attributed to the accuracy of the analytical techniques, sample preparation methods, and insufficient quality control of results in these studies.

Conclusion

Our data indicate that there is a statistically significant gender-related difference between TE levels in thyroid tissue of females and males. The Co mass fraction is higher while the Rb and Zn mass fractions are lower in female thyroids compared with those in male thyroids. Subclinical hypothyroidism is amultietiological and multifactorial complex condition. The complete understanding of the role of inadequate levels of some TE in thyroid parenchyma in the etiology of SCH requires a global vision of their different mechanisms of action, which is not yet possible with the present state of knowledge. However, from the results of our study it follows that an involvement of inadequate contents of intra-thyroidal Co, Rb and Zn in the etiology of female SCH may be assumed.

Acknowledgements

Authors are grateful to Dr. Yu. Choporov, Head of the Forensic Medicine Department of City Hospital, Obninsk, for supplying thyroid samples.

Conflict of Interest

There is no any financial interest or any conflict of interest.

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@article{vladimir2018,
  title   = {Possible Role of Inadequate Quantities of Intra-Thyroidal Cobalt, Rubidium and Zinc in the Etiology of Female Subclinical Hypothyroidism},
  author  = {Vladimir Zaichick1* and Sofia Zaichick 1, 2},
  journal = {Women\'s Health Science Journal},
  year    = {2018},
  volume  = {2},
  number  = {1},
  doi     = {10.23880/whsj-16000108}
}
Vladimir Zaichick1* and Sofia Zaichick 1, 2 (2018). Possible Role of Inadequate Quantities of Intra-Thyroidal Cobalt, Rubidium and Zinc in the Etiology of Female Subclinical Hypothyroidism. Women's Health Science Journal, 2(1). https://doi.org/10.23880/whsj-16000108
TY  - JOUR
TI  - Possible Role of Inadequate Quantities of Intra-Thyroidal Cobalt, Rubidium and Zinc in the Etiology of Female Subclinical Hypothyroidism
AU  - Vladimir Zaichick1* and Sofia Zaichick 1, 2
JO  - Women's Health Science Journal
PY  - 2018
VL  - 2
IS  - 1
DO  - 10.23880/whsj-16000108
ER  -