1.The study of allergen of mare’s milk, airag and mugwort pollen
Oyuntsatsral B ; Gantulga B ; Munkhbayarlakh S ; Narantsetseg L
Mongolian Medical Sciences 2015;173(3):19-22
BACKGROUND: Mugwort is the important source of fall allergic symptoms in the Mongolia. Mugwort pollen allergicpatients frequently present allergic symptoms of ingestion after several kinds of foods.OBJECTIVE: We sought to study the clinical manifestations of airag (fermented mare’s milk) and mare’s milkhypersensitivity in patients with mugwort allergy, identify the molecular weight of allergens, andevaluate their IgE cross-reactivity.MATERIAL AND METHODS: We collected to mugwort pollens from around UB in august and practiced fresh airag and mare’s milk.Airag, mare’s milk and mugwort extracts prepared by Hames Richmond method and their allergenswere identified by means of SDS-PAGE. ELISA inhibition experiments were done to study crossreactivitybetween airag and mugwort.RESULTS: In SDS-PAGE determined mugwort allergen 12-43 kDa, mare’s milk allergen 13-70 kDa, airag allergen12-68 kDamolecular weight.The study of the cross-reactivity between mugwort allergens and some food allergens was becamepractical significance on diagnosis, treatment and prevention of respiratory allergies. We were definedallergenic cross-reactivity between airag allergens and mugwort allergens which are common causesof upper respiratory allergy. On the Mugwort-ELISA inhibition test, the 50 % inhibitory dose to MugwortspecificIgE was 0.01μg/ml Mugwort allergens and 0.025μg/ml Airag allergens.However, the 1.0 μg/mlof Mugwort and Airag allergens were completely inhibited to Mugwort-specific IgE and Airag-specificIgE antibodies.CONCLUSION: In determined the mugwort pollen has 5 band(12, 23, 28, 38, 43kDa), in mare’s milk (13, 15, 60, 70кДа) and airag(12, 30, 50, 68 кДа) has separately 4 bands allergen protein by SDS-PAGE. ELISAinhibition study was strong cross-reactivity between airag allergen and mugwort allergen.
2.Changes follicule stimulating hormone in ageing
Batnaran D ; Solongo E ; Enebish D ; Munkhtsetseg J ; Narantsetseg L
Mongolian Medical Sciences 2010;152(2):2-5
INTRODUCTION: Gonadotropins are released under the control of gonadotropin-releasing hormone (GnRH) from thearcuate nucleus and preoptic area of the hypothalamus. The gonads — testes and ovaries — are the primary targetorgans for LH and FSH. The gonadotropins affect multiple cell types and elicit multiple responses from the targetorgans. As a simplified generalization, LH stimulates the Leydig cells of the testes and the theca cells of the ovaries to produce testosterone (and indirectly estradiol), while FSH stimulates the spermatogenic tissue of the testes andthe granulosa cells of ovarian follicles.Reproductive aging is endocrinologically characterized by a progressive rise in serum FSH levels associated with adecrease in serum estradiol (E2) and testosterone (T) levels. The rise in FSH is associated with reduced levels of sexsteroid and peptide negative feedback regulators of FSH secretion.The aim of study is: determination of serum FSH level changes in relation to aging and sex.MATERIALS AND METHODS: In this study were involved 169 healthy Mongolian adults aged above 45 years old. Subjectswere randomly selected and undergone physical examination by geriatrician. People, who are receiving hormonereplacement therapy, using inproper use of alcohol, injured and had surgery were excluded from our surgery. Bloodsamples were collected in the early morning (8.30–10.30 AM) after an all night fast and plasma was separatedimmediately by centrifugation; then sera obtained were stored at -20°C until assayed by ELISA kit from United BiotechCoLTD, USA, which sensitivity is 1mIU/ml. Statistical analyses have been performed by statistical software SPSS 16,using ANOVA, Pearson correlation, T-test.RESULT: Mean level of FSH for both sexes was 21.19±16.2 mIU/ml, which is in comparison with males (12.33±10.58mIU/ml) it was comparatively higher (p<0.001) in women (29.61±16.15 mIU/ml). FSH has no correlation with aging inman (r=0.084, p>0.05), but in women it was stronger correlation (r=0.203, p<0.05). in 51-60 years age group FSH wasincreased by 56%, in 61-65 years group by 91%, in 66-70 years group it was increased 100% in comparison with until50 years age group. In older age (above 70 years) it decreased to 70% from reached concentration. ANOVA analysishas not showed significant difference between age groups.CONCLUSIONS: Average mean of FSH in old age are: 29.61±16.15 mIU/ml in women and 12.33±10.58 mIU/ml in men.Correlation with aging was observed stronger in women than in man (r=0.203, p<0.05). FSH increases with aging untilround 70 and decreases after 70 years old.
3.Immunobiological characteristic of grass pollen allergens
Narantsetseg L ; Javzandolgor N ; Enkhbayar B ; Munkhbayarlakh S
Mongolian Medical Sciences 2014;169(3):81-86
Grass pollens are one of the most important airborne allergen sources worldwide. The Poaceaefamily comprises about 9000 species, 20 species from five subfamilies are considered to be the mostfrequent causes of grass pollen allergy, and the allergenic relationships among them closely follow theirphylogenetic relationships. The allergic immune response to pollen of several grass species has beenstudied extensively over more than three decades. Eleven groups of allergens have been identified anddescribed, in most cases from more than one species. The most complete set of allergens has so farbeen isolated and cloned from Phleum pratense (timothy grass) pollen. Based on the prevalence of IgEantibody recognition among grass pollen-sensitized individuals, several allergens qualify as major, butmembers of two groups, groups 1 and 5, have been shown to dominate the immune response to grasspollen extract. Isoform variation has been detected in members of several of the allergen groups, whichin some cases can be linked to observed genetic differences. N-linked glycosylation occurs in membersof at least three groups. Carbohydrate- reactive IgE antibodies have been attributed to grass pollensensitization and found to cross-react with glycan structures from other allergen sources, particularlyvegetable foods. Another cause of extensive cross-reactivity are the group 12 allergens (profilins), whichbelong to a family of proteins highly conserved throughout the plant kingdom and present in all tissues.Members of eight allergen groups have been cloned and expressed as recombinant proteins capableof specific IgE binding. This development now allows diagnostic dissection of the immune response tograss pollen with potential benefits for specific immunotherapy.
4.The total protein and some characterization of allergenic proteins in the body of blatella germanica cockroach
Narantsetseg L ; Oyuntsatsral B ; Javzandolgor N ; Myagmarsuren D ; Munkhbayarlakh S
Mongolian Medical Sciences 2012;161(3):17-19
Introduction: Prevalence of asthma is increasing year by year, especially among children and exposure to high levels of indoor allergens is a very important factor [1]. Cockroaches are an important cause of asthma in many other regions of the world, including Taiwan, Thailand and Singapore in the Pacific Rim, Costa Rica and Puerto Rico in Centrel America, India, South Africa and more recently, Europe [2]. Goal: The aim of this study was determined total protein amounts allergenic proteins and protein bands of сockroach.Material and Methods: The сockroachs were collected in Ulaanbaatar. The allergenic protein components of the сockroach was purified by the method of Hames Richmond. The total protein of extracts was measured by the Bradford method and the protein components of cockroach were determined by the SDS-PAGE.Results: Among the 4000 known species of cockroaches, only 5 commonly inhabit homes and have the potential to contribute to indoor allergens. These include the American (periplaneta americana), German (Blattella germanica), Oriental (Blatta orientals), Smokey brown (Periplaneta fuliginosa), and brownbanded (Supella longipalpis) varieties [3]. We were defined 2,25mg/ml protein amounts (w/v) in extracts of the purified and lyophilized protein of the сockroach. We were used a standard marker 195,7; 104,0; 59,8; 41,6; 27,8; 21,1; 15,2; 6,5kd molecular weight proteins on the 13% separation gel of SDS-PAGE. On column determined protein bands with 82,3; 59,9; 55,2; 44,0; 41,6; 34,4, 22,7, 17,1 kd molecular weights.Conclusions: The сockroach was included 8 allergenic protein components between ranges of 17,1-82,3 kd molecular weights were determined in the extracts of the body Blatella germanica.
5.Change of follicle stimulating hormone and estradiol levels in aging women and their relevance
Batnaran D ; Munkhtsetseg J ; Enebish D ; Narantsetseg L
Mongolian Medical Sciences 2010;153(3):16-19
Background:In females reproductive system aging is very important. Females spend the last 1/3 period of their life in insufficiency of sex hormone of late menopause. Due to decrease of ovary function noticeable change is detected in nervous endocrine system. Although menstrual cycle hasn’t been lost amount of FSH increased. It shows decrease of reproductive function females. Therefore recently researchers define that amount of FSH increase is an important biomarker which detects in an early period of menopause. But amount of estradiol hormone is almost in normal level until the late post menopause. Goal: to study the dynamic feature of age related changes and dependence of serum FSH and estradiol level in female aging.Materials and Methods:In this study were involved 177 healthy Mongolian women aged above 35 years old. We drained 5 ml fasting vein blood at 8.00-10.00 am. Sera were separated and kept frozen until assayed by ELISA. Results:Аaverage mean of FSH was 19.84±22.6 IU/l, at the age of 35-45 1.62±3.29 IU/l, at the age of 46-55 16.39±15.39 IU/l, at the age of 56-65 31.38±33.69IU/l, at the age of 66-75 28.83±17.31 IU/l, over 75 34.52±13.94 IU/l and there was positive correlation between age and FSH levels (r=0.647, p<0.001).Average mean of estradiol of participants: at the age of 35-45 37.03±22.09 pg/ml, at the age of 46-55 21.1±16.65pg/ml, at the age of 56-65 13.7±20.07 pg/ml, at the age of 66-75 9.11±15.1pg/ml, over 75 age 1.8±1.53 pg/ml and there was inverse correlation between age and estradiol levels (r= -0.453, р<0.001).There was an inverse correlation between log E2 and FSH levels (r= -0.434, p<0.001). Multiple regression analysis shows that age (β= -0.350, p<0.001) and FSH (β=-0.222, p=0.016) had significant inverse correlation with serum estradiol level. Conclusion: For females over 35 average mean of FSH is 19.84±2.26 IU/l, average mean of estradiol hormone is 19.6±2.0pg/ml. between the mid age (35–45 years) and oldest (>75 years) groups, mean estradiol level declined by 20.6 times, whereas FSH increased by 21.3 times. It shows that this is an important marker of menopause. Multiple regression analysis shows that age (β= -0.350, p<0.001) and FSH (β= -0.222, p=0.016) had had significant inverse correlation with serum estradiol level.
6.The study of allergen’s protein of bromus inermis pollen
Narantsetseg L ; Javzandolgor N ; Oyuntsatsral B ; Khandsuren G ; Munkhbayarlakh S
Mongolian Medical Sciences 2010;153(3):23-25
Background: The prevalence and incidence of allergic rhinitis is increasing in the last years in the Asia Pacific countries and for this reason, the number of research in aeroallergen and aeropollinology increasing. It depends on changes of geography, weather and plants, pollination period of time and air pollution.Goal: The aim of this study was determined allergenic characterization of proteins detected from Bromus inermis pollen.Mаterials and Methods: To define morphologic characteristics of Bromus inermis grass pollen and allergenic protein amounts’ of pollen and protein components.- The pollen morphologic characteristics of the Bromus inermis were defined and measured by optic microscopy (Aristoplan, Leitz, Germany).- The allergenic protein components of the Bromus inermis pollen were purified by the method of Hames, Richmond- Protein contents were measured by the Bradford method- The protein components of Bromus inermis pollen were determined by the SDS-PAGEResults and discussion:The diameter of the B.inermis dry pollen were mean length 41, 5±2, 3 μm and mean wide 32, 3±4, 1 μm. B.inermis dry pollen has oval and sphere shape and concaved on 3 sides with diameter 32.3-41.5 μm and was similar results one of subfamily of the Gramineae, Poaceae pollen size were defined 22-80 μm in diameter and with oval and sphere shapes. We were defined 1.5±0.02 mg protein amounts in the 5mg/ml extracts of the purified of the Bromus inermis pollen. Researcher [3] determined 1, 45 mg/ml protein on Elymus chinensis, 1, 96 mg/ml protein on Artimesia sieversiana, 3, 29 mg/ ml protein on Chenopodium album allergens. These study results are similar with our study result on Bradford method. We were defined 7 bands with 12, 26, 32, 55, 66, 84, 97 kDa molecular weight protein components. SDS-PAGE were deteсted relatively bright bands of 12, 32, 55, 66 kDa molecular weight protein components of Bromus inermis pollen proteins. Researcher Kaiser M et al were defined 16, 30, 40, 47, 50, 57, 60, 67, 70, 90, 95 and 110 kDa molecular weight bands in Lolium perenne pollen allergens. These study results are similar with our study result on SDS-PAGE. Conclusions:- The pollen of Bromus inermis was oval and sphere shapes with 32.3-41.5 μm in diameter.- We were defined 1.5±0.02 mg protein amounts in the 1mg/ml of the Bromus inermis pollen.- The 7 bands with 12, 26, 32, 55, 66, 84, 97 kDa molecular weight protein components of Bromus inermis pollen. SDS-PAGE were deteсted relatively bright bands of 12, 32, 55, 66 kDa molecular weight protein components.
7.The study of co sentization allergens to pollen of dandelion (taraxacum), pyrethrum (aquilegia and) weed, grass plants
Nyamdavaa N ; Janerke T ; Enkhbayar B ; Munkhbayrlakh C ; Narantsetseg L
Mongolian Medical Sciences 2015;172(2):87-90
IntroductionOver the last few years, the prevalence of aeroborn allergic disease is rising rapidly throughoutdeveloped and developing countries, and one of every four children in Western Europe has had allergy.In any country of the world over the last 10-20 years, studies on airopollinology and aeroallergenshowing increase of allergen rhinitis to pollen. This is catching many scientists and researchersattentions. These studies deemed that geological locations, climate and plant patterns changing as airpollution increases. Studies identified that pollen allergy in Europe is mainly produced by segmentedplants and in north Europe Betulacease type of allergy is dominating [2]. In our country, Mongolia,plant sturctures are relatively well studied geologically and climatically, and 203 species of plants,98 types and 31 families showed plants may cause allergies. Specifically recent years in Umnugoviprovince, with the expanding operations of many mining companies, increased air pollution leadingto a respiratory disease. With the uses of Compositae in medicine, cosmetics and food, sentization toplant of this family has been increasing in Europe as well as in Asia [3].Goal:To determine co sentization allergens to pollen of Taraxacum, columbine and weed, grass plants.Materials and МethodsThe Research has been done under the Biochemistry Department of Bio – Medical School, HSUMwith the help of “Effect” Allergy – Asthma Hospital. During the study of research, one period descriptiveresearch is done by studying the selected 432 patients who are diagnosed positive for the TaraxacumandAquilegia allergens by skin pricking test and these group is chosen from the airborne allergic patients“Effect” Allergy – Asthma Hospital in 2010 -2014 census.Tables and graphs showing study result were processed by using Excel-2013, SPSS-21.0 software.ResultIn this study, in 2010-2014, aeroallergens skin prick test was done on 5601 people and 18% or 1031people were aeroallergen sensitized.ConclusionsFindings from the skin prick tests for airborne allergens show that 8%, 5% of the patient is positive forTaraxacum, Aquilegia allergen. On SPTs taraxacum, aquilegia and mugwort 60% and grass pollen56,90% of population was cosensitized to all pollens.
8.The result of rongalite determination in some fruit grown in Mongolia
Byambasuren D ; Bayarmaa E ; Bayarjargal N ; Narantsetseg L
Health Laboratory 2017;6(1):24-26
Introduction:
Rongalite have many names, the trade name for sodium hydroxymethylsulfinate also sodium formaldehydesulfoxylate. It causes serious side effects to human body and is forbidden to be used as food additives by law. It is still frequently used illegally in rice and flour products. Based on this study, we determination rongalite level in some friut widespread of in our country. The study was done materials used in Lingonberry, Blueberry, Hippophae, Blackcurrant fruits collected in Khuvsgul. To determine rongalite was followed by protocol to sanitation-laboratory of food of the school of social of health in MNUMS used for NC-860 Universal Food Safety Analyzer. The concentration in sample that were measure to 153.88- 213.85 mg/kg of Lingonberry, Blueberry, Hippophae, Blackcurrant fruits weight.
Materials and methods :
The study was done materials used in a Lingonberry, Blueberry, Hippophae, Blackcurrant fruits collected in Khuvsgul. To determine rongalite was followed by protocol to sanitation-laboratory of food of the school of social of health in MNUMS used for NC-860 Universal Food Safety Analyzer. Weigh accurately 5.0 g of the fruit in a plastic extract bottle, add 20 ml of distilled water, extract for 10 min by centrifugated. Than filtrate into 6 time respectively test tubes through 0.45 μm membrane using remove the color. Rongalite determined in use a disposable test tube, add 1 drop of oxidizing reagent. Amount of rongalite used for NC-860 Universal Food Safety Analyzer.
Result and discussion
Based on the obtained result, the maximum concentration observed in Blueberry was 213.85 mg/kg, and the minimum concentration observed in Hippophae was 153.88 mg/kg, the concentration observed in Lingonberry was 192.8 mg/kg, the concentration observed in Blackcurrant was 155.29 mg/kg of contained weight respectively. The difference of rongalite concentrations in wheat powder and rice powder was 0.41 mg/kg in China. Identified to the result was higher than other research.
9.ISOLATION AND PURIFICATION OF IMMUNE MODULATING LACTOFERRIN FROM MONGOL BOVINE COLOSTRUM
Chingunjav E ; Jambal B ; Amarsaikhan B ; Gerelmaa T ; Narantsetseg L ; Sarantuya R ; Bilegtsaikhan Ts ; Purevjargal N ; Tengis A ; Javkhlan B ; Tsendmaa Ts ; Galindev B ; Munkhtulga L ; Nyambayar D ; Munkhbat B ; Baigalmaa B
Innovation 2017;11(1):30-33
BACKGROUND
Bovine colostrums is the milk secreted by cows during the first few days after parturition. It
contains many essential nutrients and bioactive components, including growth factors,
immunoglobulins, lactoperoxidase, lactoferrin and cytokines ets. Lactoferrin has been reported
for its multifunctional properties such as antifungal, antibacterial, antiviral antioxidant and
anticancer activities. The aims of this study focused on the isolation and purification of lactoferrin
from Mongolian bovine colostrums. Lactoferrin purified using HiTrap DEAE an ion exchange
chromatography. Lactoferrin purification efficiency was about 60.5%. The single band of purified
lactoferrin has been observed in SDS-PAGE electrophoresis.
METHODS
Bovine colostrum was collected at a cow farm in the Darkhan province of Mongolia. At first
the cream was separated by centrifugation (10000 xg 20 min at 4oC). In order to separate the
whey, the samples were precipitated with 1mol/l to pH 4.6 and centrifuged at 10000 g 20 min
again. The samples of whey were stored at -18oC to the analysis. Lactoferrin was purified by
HiTrap DEAE an ion exchange chromatography using 0.005 M phosphate buffer (pH 7.7) and
linear gradient NaCl from 0.25M, 0.5M, 1M. During chromatography, protein in the eluents was
monitored by ultraviolet absorbation at 280 nm with the instrument. Purity test done by using
polyacrylamide gel electrophoresis under denaturated condition (SDS-PAGE) method by Laemmli
(1970). For HPLC determination of the lactoferrin by Shimadzu Nexera X2 HPLC system with UV/
VIS detector were used. Detection was carried out at the wavelength 280 nm. Separation was
performed on a chromatographic column Protein R C18 ,2.2 x 150 mm, 5 μm particle size. Linear
gradient and flow rate 0.2 ml/min were used. Mobile phase a consisted of water / acetonitrile/
trifluoroacetic acid ( 95:5:0.1). The column temperature was set at 40oC and injection volume
was 10 μl. Data were collected and evaluated by software Lab Solution. An external standard
method for quantification analytes was used.
RESULTS
Purified lactoferrin in the present study had a good concentration and purification efficiency
was about 60.5 %. Protein fraction from 1M NaCl gradient delivers sharp and clean peak to
HPLC chromatogram that fits intensity and retention time of standard bovine lactoferrin.
Ammount of lactoferrin in bovine colostrums was 0.6 mg/ml and it`s molecular weight 80 kDa as
a standard sample. The retention time of lactoferrin fraction which is purified by SDS-PAGE gel
electrophoresis. The peak of fraction same compared to the standard lactoferrin 5.8 minutes
by HPLC analysis.
CONCLUSION
Ion exchange chromatography shows reliable and easy isolation of lactoferrin from Mongol
bovine colostrum.