Semipalatinsk nuclear test site (STS) (Kazakhstan)


      The Semipalatinsk Nuclear Test Site (STS) also known as gPolygonh is located on northeast Kazakhstan being attached to the Lrtysh river. The STS has been a primary venue for the nuclear test site of the former Soviet Union. Since the first nuclear test on September 1949, about 456 test explosions were performed until closure of STS on August 29, 1991. All together approximately one million residents of the villages near the test site were estimated to be exposed to radioactive fallout, mostly from the fallout of atmospheric explosions performed until 1962. The explosions thereafter were underground explosions. Hydrogen fusion explosion performed (-178m underground explosion with yield of 140 kt TNT equivalent) on January 15, 1965 resulted in a large lake of about 400 m diameter and 100 m depth (Chagan Lake; i.e., also known as gatomic lakeh).


[1] Preliminary study on chromosome aberrations, GPA- and TCR-mutation at STS (RBC, Kyoto University) 

     Blood samples were collected from residents of Kurchatov, Akzhar and Sarzhal on May 16-19, 2000. Blood samples were immediately mixed with equal amount of gcold stimulating medium*h and kept in the cool (below 20 oC). The blood samples were brought by plane to RBC, Kyoto University, where lymphocyte cultures were established on May 26, 2000 for chromosome aberration analysis, and a part of the blood sample (still in the cold stimulating medium kept in the cold) was mailed to Radiation Effects Research Foundation, where GPA and TCR mutations were analysed by Dr. S. Kyoizumi and Dr. T. Seyama. The results are presented elsewhere.

Commentary
     The cold stimulation method, which has been introduced in IAEA Technical Reports in 2001 and 2011. The method has been devised to keep the lymphocytes viable for a long time after blood sampling. However, unfortunately the method has not been applied as originally recommended, in particular, some reports use regular culture media, such as RPMI, and attempt to keep nutritional condition and pH by adding L-glutamine and HEPES. The original protocol recommends the use of Leibovitzfs L-15 medium. This culture medium is the one specifically produced to culture in ambient atmosphere without aeration of CO2 gas. The L-15 medium is composed of 10 times amounts of amino acids, i.e., the amino acid-based buffered against acidification by respiration of red blood cells, and high supply of nutrition as well. The use of other commercially available culture media resulted in earlier death of lymphocytes.
     Sasaki, M. S. (1998): Pulling lymphocyte viability for long-term transportation of blood samples. IAEA Symposium on Cytogenetic Analysis for Radiation Dose Assessment. June 29-July 2, 1998. Budapest, Hungary.
     IAEA (2001): Cytogenetic Analysis for Radiation Dose Assessment: A Manual. IAEA Technical reports series No. 405. International Atomic Energy Agency.
    IAEA (2011): Cytogenetic Dosimetry: Applications in Preparedness for and Response to Radiation Emergencies. International Atomic Energy Agency.



[2] Residents of Dolon (Testa et al. 2001) 
Reference  

     Testa, A., Stronati, L., Ranaldi, R., Spano, M., Steinhausler, F., Gastberger, M., Hubmer, A., Ptitkaya, L. and Akhmetov, M. (2001): Cytogenetic biomonitoring carried out in a village (Dolon) adjusent to the Semipalatinsk nuclear weapon test site. Radiat. Environ. Biophys., 40:125-129. 
Study design 

      Blood samples were collected from inhabitants of Dolon who have been living since 1949 (time of the first test explosion). The blood samples were immediately placed into polyform boxes and transferred by plane to the ENEA laboratory in Italy (with 36 hrs). Whole blood cultures were established, and chromosome preparations were stained with Giemsa. Control samples were collected from personnel of the Institute of Radiation Safety and Ecology of Kuruchatov. They were mainly involved in the administrative works and any occupational radiation exposure could be exclude.
Observations 
Prsons living in contaminated area. Persons living in non-contaminated area.
Subject Age Sex No. of Aberrations Subject Age Sex No. of Aberrations
ID (year) cells Dic+cR Ace ID (year) cells Dic+cR Ace
a 47 M 200 2 0 1 57 F 200 1 8
b 50 M 200 1 1 2 60 M 200 0 1
c 62 F 200 0 4 3 60 M 200 0 0
d 59 F 200 0 0 4 58 F 200 0 1
e 66 F 200 0 3 5 55 F 200 0 2
f 57 F 200 0 0 6 49 F 200 0 0
g 74 F 200 0 0 7 48 F 200 0 1
h 50 M 200 2 0 8 58 F 200 1 3
i 70 M 200 1 3 9 47 F 200 0 0
l 49 F 200 1 2 10 44 F 200 0 0
m 68 M 200 1 2 11 48 F 200 0 1
n 61 F 200 0 2 12 50 M 200 0 2
o 54 F 200 0 0 13 54 F 200 0 0
p 74 F 200 1 5 14 66 M 200 0 0
q 67 F 200 0 1 15 47 200 0 0
r 66 F 200 1 4 16 46 F 200 0 0
s 65 M 200 0 0 17 78 F 200 1 1
t 55 F 200 0 1 18 60 M 200 0 0
u 68 F 200 0 3 19 57 F 200 0 2
v 64 F 200 0 2 20 36 F 200 0 0
z 67 M 200 1 2
 

The distributions suggest difference between the exposed and non-exposed groups.



[3] Residents of areas near the STS (Tanaka et al. 2006) 
Reference  

     Tanaka, K., Iida, S., Takeichi, N., Chaizhunusova, N. J., Gusev, B. I., Aspalikov, K. N., Inaba, T. and Hoshi, M. (2006): Unstable-type chromosome aberrations in lymphocytes from individuals living near Semipalatinsk nuclear test site. J. Radiat. Res., 47(suppl):A159-A164. 
Study design 

     Blood samples were obtained from healthy residents of differently contaminated areas near STS, Dolon, Sarjar, Kaynar, Znamenka, Semipalatinsk and Socialistic, and remotely located uncontaminated area, Kokpekty. The residents had been living in the areas between 1948 and 1965 when the atmospheric nuclear tests were performed.
     Blood samples were transferred to Japan, where two series of lymphocyte cultures were established: one for chromosome aberration analysis and another for micronucleus assay. For chromosome aberration analysis, whole blood cultures were established, incubated for 52 hours. Chromosome preparations were stained with Giemsa. For micronucleus assay, lymphocytes were isolated and cultured for 72 hours with last last 28 hours in the presence of cytochalasin B. Micronuclei were counted in the Giemsa stained binucleated cells. 
Observations 
[1] Unstable-type chromosome aberrations [2] Micronuclei (MN)
Villages No. of Age No. of No. of Aerrations No. of No. of No. of cells Mean no. of No. of MN
subjects (mean}SD) cells abnormal cells Dic* cR subjects cells (mean) with MN (mean) MN/subject per 1000 cells
Exposed Dolon 35 58.5}5.18 9,794 62 17 (9) 8 53 5,018.5 46.8 50.8 9.36}3.46
Sarjar 48 56.9}5.36 13,642 141 16 (6) 7 44 4,098.3 38.9 42.2 9.9}3.62
Kaynar 33 56.9}4.84 11,650 40 14 (8) 4 43 5,132.4 50.2 55.8 9.87}3.62
Znamenka 3 49.7}0.57 1,340 41 5 (0) 4 10 5,112.1 35.9 35.9 7.1}3.0
Semipalatinsk 4 51.3}1.26 803 21 2 (0) 2 42 4,734.2 61.8 61.8 12.3}3.94
Socialistic - - - - - - 20 4,348.4 31.8 34.1 7.3}3.08
Controls Kokpekty 46 52.1}3.34 14,192 66 9 (6) 2 21 5,034.5 36.5 38.5 7.25}2.14
*) Number in parentheses shows dicentrics with asssociated fragments.
**) Higher aberration frequencies were found in some of the contaminated areas, but none was statistically significant as compared to the values in the control populations.


[4] Residents of Dolon (Stephan et al. 2001)
Reference 
     Stephan, G., Pressl, S., Koshpessova. And Gusev, B. I. (2001): Analysis of FISH-painted chromosomes in individuals living near the Semipalatinsk nuclear test site. Radiat. Res., 155:796-800.  
Study design 
     Blood samples were collected from 10 subjects who were born before the first explosion in 1949 and lived continuously in the village of Dolon. Dolon is highly polluted area among other villages.
hromosome translocations were analyzed by FISH painting method using whole chromosome painting probes for chromosomes 2, 4 and 8.
     The external dose to the inhabitants was estimated based on the physical measurements of dose rates along the track of radioactive clouds from explosion. Data derived from radiochemical analyses of soil, water, vegetation and food samples were used to estimate the internal dose. In Dolon, the effective external dose contributed about 48 % and the internal dose about 52 % of the total effective equivalent dose (B. J. Gusev, personal communication cited therein). 

Observations 
Subject Age* Effective equivalen dose (Sv) No. of Chromosome aberrations** Cells with
ID (years) External Internal Total cells tc ti dicc dici del cr complex aberrations
KD1 0.2 1.61 1.72 3.33 2,484 3 4 1
KD2 2.8 1.61 1.73 3.34 2,073 6 1 3 2
KD3 2.0 1.61 1.73 3.34 2,172 13 3 2 2 1
KD4 12.2 1.61 1.44 3.05 2,005 3 1 3a
KD5 2.2 1.61 1.73 3.34 2,121 4 1 1
KD6 1.4 1.61 1.73 3.34 2,455 2 6b 1 1 3
KD7 2.9 1.61 1.73 3.34 2,192 2 1 1
KD8 4.3 1.61 1.73 3.34 2,555 2 2c 1 1
KD9 3.3 1.61 1.73 3.34 2,123 3 1 3
KD10 3.2 1.61 1.73 3.34 2,060 2 1
*) Age: age at the first explosion in 1949.
**) t: translocation. dic: dicentrics. del: deletions. cr: centric rings. Subscripts (c) and (i) are "complete" and "incomplate" exchanges, respectively.
a): One cell containing t(Ba)+t(Ab)+2t(Ab)+dic(AB); one cell with all labelled chromosome damaged.
b) One translocation in combination withone dic and one t(bAA).
c) One translocation in combination with one ring.


[5] Residents of districts near the STS (Salomaa et al. 2002)
Reference 

     Salomaa, S., Lindholm, C., Tankimanova, M. K., Zh. Mamyrbaeva, Z., Koivistoinen, A., Hulten, M., Mustonen, R., Dubrova, Y. E. and Bersimbaev, R. I. (2002): Stable chromosome aberrations in the lymphocytes of a population living in the vicinity of the Semipalatinsk nuclear rest site. Radiation Res., 158:591-596. 
Study design 

     The frequencies of stable-type chromosome aberrations have been studied by FISH chromosome painting technology in the inhabitants of villages near the Semipalatinsk nuclear test site (STS).
To test the validity of the use of stable translocations for retrospective biological dosimetry, the study populations were categorized into two groups; members of P0 generation who were living from the first nuclear test explosion in 1949, and members of P1 generation who were bone between 1955 and 1985. Because the massive atmospheric explosion were carried out between 1949 and 1956 (1957?), the P0 generation is expected to have been exposed to high doses from fallout as compared to P1 generation. They were all residents living in areas of Beskaragai district contaminated by fallout from STS. Further to compare any effects, the non-contaminated controls were selected from the Taldy Kurgan district in southeast Kazakhstan.
     Blood sampling was performed within a year of 1999. Lymphocytes were isolated and cultured at the Institute of General Genetics and Cytology in Almaty. The cultures were matched for 60 exposed and 40 control subjects for age, generation, smoking and ethnic background. FISH painting was performed by the use of whole chromosome painting probes for chromosomes 1, 2 and 4. 
Conclusion 
     The translocation frequencies were not significantly different among P0, P1 generations living in contaminated district and controls living in non-contaminated district. The observations could not substantiate the previously reported does of the order of 1-4.5 Gy to P0 generation in the contaminated area. Moreover, the age-dependent increase of translocation frequencies was not diferent between the exposed and control populations. 
Generations FISH analysis Village exposed Controls
Bodene Chagan Cheremushki Dolon Kanonerka Karamyrza Mostik All combined
P0 No. of subjects 8 1 2 10 8 1 1 31 19
No. of cells analysed 15,800 2,000 3,860 19,243 13,752 1,105 753 56,513 40,743
Two-way translocations* 8.7}1.3 5.7}2.8 7.7}2.3 9.5}1.2 4.8}1.0 5.2}3.7 3.8}3.8 7.7}0.6 8.1}0.8
All translocations* 10.8}1.4 8.6}3.5 8.9}2.6 11.9}1.5 8.1}1.3 5.2}3.7 7.6}5.4 10.0}0.7 10.2}0.8
P1 No. of subjects 12 - 3 3 10 - - 28 21
No. of cells analysed 24,276 - 7,231 6,000 18,029 - - 55,536 43,999
Two-way translocations* 1.5}0.4 - 4.7}1.4 2.4}1.1 4.0}0.8 - - 2.8}0.4 5.8}0.6
All translocations* 2.0}0.4 - 7.9}1.8 2.4}1.1 5.7}0.9 - - 4.0}0.5 5.8}0.6
*) Translocations per 1000 cell equivalent } SE. Two-way translocation: t(Ab)+t(Ba). Other translocation: t(Ab) or t(Ba).





[6] Residents of Dolon and Chekoman (Chaizhunusova et al. 2006) 
Reference  

     Chaizhunusova, N., Yang, T. C., Land, C., Luckyanov, N., Wu, H., Apsalikov, K. N. and Madieva, M. (2006): Biodosimetry study in Dolon and Chekoman villages in the vicinity of Semipalatinsk nuclear test site. J. Radiat. Res., 47 (suppl):A165-A169. 
Study design 

     A total of 15 women were selected from Dolon and 15 women from Chekoman districts. They were all born before the first nuclear test in August 1949. Chromosome aberrations in blood lymphocytes were studied by FISH painting technique.
     Chromosome aberrations were higher in the resident of Dolon as compared to those in Chekoman, indicating a reflection of the estimated dose levels of the two districts. 
Observations 
[1] Dolon [2] Chekoman
Subject Cells Aberrations Subject Cells Aberrations
ID scored Number Genome equivalent ID scored Number Genome equivalent
D1 964 8 26.4 C1 851 1 3.3
D2 221 0 0 C2 2011 5 16.5
D3 3348 11 36.3 C3 283 0 0
D4 3562 9 29.7 C4 934 1 3.3
D5 1105 5 16.5 C5 712 0 12.4
D6 132 1 3.3
D7 181 0 0
D8 164 1 3.3
D9 58 1 3.3
D10 2723 19 62.7