Luksana Makonkawkeyoon

Helicobacter pylori infection in combination with the serum pepsinogen I/II ratio and interleukin-1β-511 polymorphisms are independent risk factors for gastric cancer in Thais

BackgroundThailand has the lowest incidence of gastric cancer in the world. Helicobacter pylori infection, a low serum pepsinogen I/II ratio, and interleukin (IL)-1β-511 polymorphisms are suspected to be risk factors for gastric cancer.MethodsA total of 167 Thais, comprising 56 cancer patients and 111 volunteers without cancer, underwent an esophagogastroduodenoscopic examination and three fixed-point biopsies; a cancer tissue biopsy was also done, and blood samples were collected. The subjects without cancer were divided into normal subjects and chronic gastritis patients. IL-1β-511 polymorphisms were analyzed by polymerase chain reaction-restriction fragment length polymorphism, and the serum levels of pepsinogen I and II were determined by a radioimmunoassay. Helicobacter pylori IgG antibody and tissue pathology were tested in all groups.ResultsThe pepsinogen I/II ratio was significantly lower in the gastric cancer group than in the normal and chronic gastritis groups [odds ratio (OR), 2.3; 95% confidence interval (CI), 1.10–4.80; P = 0.025]. Gastric cancer patients were positive for the H. pylori IgG antibody more frequently than negative (OR, 2.946; 95% CI, 1.4–6.39; P = 0.005). However, only 15 (27%) cancer patients were both positive for H. pylori IgG antibody and had low serum pepsinogen I/II. The C/C genotype was found more frequently in the gastric cancer group than in the group with a normal gastric mucosa (OR, 0.64; 95% CI, 0.50–0.81; P = 0.014).ConclusionsA low serum pepsinogen I/II ratio combined with positivity for H. pylori IgG, and a IL-1β-511 C/C genotype may be independent risk factors for gastric cancer in Thais.

Ginger extract inhibits human telomerase reverse transcriptase and c-Myc expression in A549 lung cancer cells.

The rhizome of ginger (Zingiber officinale Roscoe) has been reputed to have many curative properties in traditional medicine, and recent publications have also shown that many agents in ginger possess anticancer properties. Here we show that the ethyl acetate fraction of ginger extract can inhibit the expression of the two prominent molecular targets of cancer, the human telomerase reverse transcriptase (hTERT) and c-Myc, in A549 lung cancer cells in a time- and concentration-dependent manner. The treated cells exhibited diminished telomerase activity because of reduced protein production rather than direct inhibition of telomerase. The reduction of hTERT expression coincided with the reduction of c-Myc expression, which is one of the hTERT transcription factors; thus, the reduction in hTERT expression might be due in part to the decrease of c-Myc. As both telomerase inhibition and Myc inhibition are cancer-specific targets for cancer therapy, ginger extract might prove to be beneficial as a complementary agent in cancer prevention and maintenance therapy.

Predominant mucosal IL-8 mRNA expression in non-cagA Thais is risk for gastric cancer.

AIM To study gastric mucosal interleukine-8 (IL-8) mRNA expression, the cytotoxin-associated gene A (cagA) mutation, and serum pepsinogen (PG) I/II ratio related risk in Thai gastric cancer. METHODS There were consent 134 Thai non-cancer volunteers who underwent endoscopic narrow band imaging examination, and 86 Thais advance gastric cancer patients who underwent endoscopic mucosal biopsies and gastric surgery. Tissue samples were taken by endoscopy with 3 points biopsies. The serum PG I, II, and Helicobacter pylori (H. pylori) immunoglobulin G (IgG) antibody for H. pylori were tested by enzyme-linked immunosorbent assay technique. The histopathology description of gastric cancer and non-cancer with H. pylori detection was defined with modified Sydney Score System. Gastric mucosal tissue H. pylori DNA was extracted and genotyped for cagA mutation. Tissue IL-8 and cyclooxygenase-2 (COX-2) mRNA expression were conducted by real time relative quantitation polymerase chain reaction. From 17 Japanese advance gastric cancer and 12 benign gastric tissue samples, all were tested for genetic expression with same methods as well as Thai gastric mucosal tissue samples. The multivariate analysis was used for the risk study. Correlation and standardized t-test were done for quantitative data, P value < 0.05 was considered as a statistically significant. RESULTS There is a high non cagA gene of 86.8 per cent in Thai gastric cancer although there are high yields of the East Asian type in the positive cagA. The H. pylori infection prevalence in this study is reported by combined histopathology and H. pylori IgG antibody test with 77.1% and 97.4% of sensitivity and specificity, respectively. The serum PG I/II ratio in gastric cancer is significantly lower than in the non-cancer group, P = 0.045. The serum PG I/II ratio of less than 3.0 and IL-8 mRNA expression ≥ 100 or log10 ≥ 2 are significant cut off risk differences between Thai cancer and non-cancer, P = 0.03 and P < 0.001, respectively. There is a significantly lower PGI/II ratio in Japanese than that in Thai gastric cancer, P = 0.026. Serum PG I/II ratio at cut off less than 3.0 and IL-8 mRNA expression Raw RQ > 100 or log10 > 2 are significantly difference between Thai cancer group when compared to non-cancer group, P = 0.013 and P < 0.001, respectively. In the correlation study, low PG I/II ratio does not associate with chronic atrophic gastritis severity score in Thais non-cancer cases. However, there is a trend, but not significant convert correlation between IL-8 mRNA expression level and low PG I/II ratio in Thai positive H. pylori infection. The high expression of IL-8 gene demonstrates a poorer prognosis by stage and histology. CONCLUSION Predominant gastric mucosal IL-8 mRNA expression level, H. pylori infection, and low PG I/II ratio are relative risks for Thai gastric cancer without correlation with cagA mutation.

Comparison of seasonal variation in the fasting respiratory quotient of young Japanese, Polish and Thai women in relation to seasonal change in their percent body fat

BackgroundFrom the viewpoint of human physiological adaptability, we previously investigated seasonal variation in the amount of unabsorbed dietary carbohydrates from the intestine after breakfast in Japanese, Polish and Thai participants. In this investigation we found that there were significant seasonal variations in the amount of unabsorbed dietary carbohydrates in Japanese and Polish participants, while we could not find significant seasonal variation in Thai participants. These facts prompted us to examine seasonal variations in the respiratory quotient after an overnight fast (an indicator of the ratio of carbohydrate and fat oxidized after the last meal) with female university students living in Osaka (Japan), Poznan (Poland) and Chiang Mai (Thailand).MethodsWe enrolled 30, 33 and 32 paid participants in Japan, Poland and Thailand, respectively, and measurements were taken over the course of one full year. Fasting respiratory quotient was measured with the participants in their postabsorptive state (after 12 hours or more fasting before respiratory quotient measurement). Respiratory quotient measurements were carried out by means of indirect calorimetry using the mixing chamber method. The percent body fat was measured using an electric bioelectrical impedance analysis scale. Food intake of the participants in Osaka and Poznan were carried out by the Food Frequency Questionnaire method.ResultsThere were different seasonal variations in the fasting respiratory quotient values in the three different populations; with a significant seasonal variation in the fasting respiratory quotient values in Japanese participants, while those in Polish and Thai participants were non-significant. We found that there were significant seasonal changes in the percent body fat in the three populations but we could not find any significant correlation between the fasting respiratory quotient values and the percent body fat.ConclusionsThere were different seasonal variations in the fasting respiratory quotient values in the three different populations. There were significant seasonal changes in the percent body fat in the three populations but no significant correlation between the fasting respiratory quotient values and the percent body fat.

Production of a mouse hybridoma secreting monoclonal antibody highly specific to hemoglobin Bart's (gamma4).

Hemoglobin (Hb) Barts (gamma4) was isolated and purified from Hb Barts hydrops fetalis syndrome blood by CM-Sephadex C-50 chromatography. The isolated Hb Barts was analyzed for its purity by high performance liquid chromatography. Balb/c mice were immunized intraperitoneally with Hb Barts. The immunized mouse splenic cells were hybridized with mouse myeloma, X63-Ag8.653, by polyethylene glycol. There were 12 hybridoma clones, out of several thousand culture wells, secreting antibody against purified Hb Barts. However, when those 12 monoclonal antibodies (mAb) were tested with Hb Barts (gamma4), HbF (alpha2gamma12), HbH, HbE, and HbA2, there was only 1 hybridoma clone secreting mAb highly reactive to Hb Barts with very low reactivity to HbF. A rabbit polyclonal antibody with relative high reactivity to Hb Barts compared to HbF with the ratio of 2.4:1 was also produced by affinity column chromatography for the purpose of developing an enzyme-linked immunosorbent assay (ELISA) base for qualitative and quantitative determination of Hb Barts in adult hemolysates. Preliminary results in quantitative determination of Hb Barts in Hb solution of 3 alpha thalassemia families having at least 1 child with HbH disease and 6 normal subjects indicated that it was possible to quantify Hb Barts by our developed ELISA with appropriate sensitivity and specificity.

Comparison of variations between percentage of body fat, body mass index and daily physical activity among young Japanese and Thai female students

BackgroundIn our series of investigations concerning the causes of seasonal change in fat accumulation in young university students, we could not find any contribution of seasonal variation in the ratio of carbohydrate and fat metabolism to that of body fat percentage in Japanese and Thai participants. After our previous study, we examined the effect of daily physical activity on body fat percentage to look for the major causes of seasonal change in fat accumulation in young university students.FindingsIn this study, we measured participants’ (young Japanese and Thai university students) daily physical activity by a uniaxial accelerometer in addition to the measurements of body fat percentage and body mass index by a bioelectrical impedance meter. We found that there was significant and moderate negative correlation between body fat percentage and daily step counts among Japanese but not Thai participants. We observed significant, moderate and positive correlations between the percentage of body fat and body mass index among Japanese and Thai participants.ConclusionsDaily physical activity plays an important role in the seasonal variation of body fat percentage of Japanese female students. Our present study also confirmed the importance of daily physical activity for controlling body mass index and for the prevention of obesity.

Development of an ELISA strip for the detection of α thalassemias

α thalassemia is probably the most common of all single-gene disorders throughout the world. Most incidences of α thalassemia arise from the deletion of one (-α) or both (--) of the α globin genes, which are known as α+ thalassemia (-α/αα) or α thalassemia (--/αα), respectively.[1][1] The

Prevalence of α-thalassaemia genotypes in pregnant women in northern Thailand

Background & objectives: Alpha-thalassaemias are genetic disorders with high prevalence in northern Thailand. However, common genotypes and current data on the prevalence of α-thalassaemias have not been reported in this region. Therefore, the objective of the present study was to determine the prevalence of α-thalassaemia genotypes in pregnant women in northern Thailand. Methods: Genomic DNA was extracted from blood samples of pregnant women who came to Maharaj Nakorn Chiang Mai University Hospital during July 2009 to 2010. The common deletion and point mutation genotypes of α-thalassaemia were evaluated by gap- polymerase chain reaction (PCR) and PCR with restriction fragment length polymorphism (RFLP). Results: Genotypes of 638 pregnant women were: 409 samples (64.11%) being normal subjects (αα/αα) and 229 samples (35.89%) with α-thalassaemias. These 229 samples could be classified into deletional HbH disease (--SEA/-α3.7) for 18 samples (2.82%); heterozygous α0-thalassaemia --SEA type (--SEA/αα)) for 78 (12.23%); heterozygous α+-thalassaemia - α3.7 type (-α3.7/αα) for 99 (15.52%); homozygous α+-thalassaemia - α3.7 type (-α3.7/- α3.7) for five (0.78%); heterozygous α+-thalassaemia - α4.2 type (-α4.2/αα) for two (0.31%); and heterozygous HbCS (αCSα/αα) for 27 (4.23%) cases. Interpretation & conclusions: The prevalence of α-thalassaemias in pregnant women in northern Thailand was high. This finding supports the implementation of the prevention and control of this common genetic disorder by screening for α-thalassaemia genotypes.

Application of monoclonal and polyclonal antibodies to Hb Bart's for the detection of α thalassemias

Using a mouse monoclonal antibody (mAb) (“2D4”) with high specific reactivity to Hb Bart’s and a rabbit polyclonal antibody (“RPB”) with high reactivity to Hb Bart’s but low reactivity to HbF, an ELISA assay was developed for the quantification of Hb Bart’s in hemolysates of peripheral blood. In the preliminary study, hemoglobin solutions containing 4,000 μg/mL of hemoglobin were analyzed for the concentration of Hb Bart’s in samples collected from the following children and adult subjects of HbH families: 12 children with deletional HbH disease (--/3.7 ) or nondeletional HbH disease (HbH disease with HbCS) (--/ cs ), 12 adults with 0 thalassemia (--/ ), 12 adults with deletional or nondeletional + thalassemia (3.7 / or / cs ) and 12 normal adult subjects ( / ). The mean ± S.D. of Hb Bart’s concentration in those with deletional HbH disease or HbH disease with HbCS, 0 thalassemia, deletional or nondeletional + thalassemia, and normal subjects were 1,374±210 (range 1,164-1,584), 1,118±357 (range 761-1,475), 451 ± 230 (range 221-681), and 0 ng/mL, respectively. When the developed ELISA was further evaluated with additional samples of various types of thalassemia, including: 18 with deletional HbH disease (--/3.7 ); 21 of nondeletional HbH disease (HbH disease with HbCS) (--/ cs ); 33 with 0 thalassemia (--/ ); 19 with nondeletional + thalassemia ( / cs ); 11 with deletional + thalassemia (3.7 / ) and 58 normal subjects ( / ). It was found that the levels of Hb Bart’s in deletional + thalassemia was significantly lower than in 0 thalassemia (p<0.001). The levels of Hb Bart’s in 0 thalassemia was also significantly lower than in nondeletional and deletional HbH diseases (p=0.023 and p<0.001, respectively). When all types of thalassemia were compared with normal subjects, the Hb Bart’s levels in all types of thalassemia were significantly higher (p<0.0001). All of our results indicated that the developed ELISA was highly sensitive and specific for quantitative determination of Hb Bart’s in hemolysates. The ELISA assay might be used as a rapid screening test for the detection of thalassemias in general population.

Abstract 1148: The factual evidence of gastric mucosal IL-8 mRNA expression, cagA gene H.pylori infection, and pepsinogen I/II ratio in Thai gastric cancer .

Background: Interleukine-8 (IL-8) gene expression was reported in vitro that it may relate to cagA gene in Japanese. There is no in vivo study demonstrated how its level related to cagA genotype and serum pepsinogen I/II ratio in other Asian Ethnic beside of East Asian such as in Japanese. We aimed to study the relative risk of the cytokine gene expression level in Thai gastric cancer and how its correlation to other co-factors Methods: There were 86 Thai gastric cancer patients and 134 Thai non-cancer volunteers who underwent endoscopic mucosal biopsies. The serum pepsinogen I, II, and Helicobacter pylori Immunoglobulin G antibody were tested. The extracted H.pylori DNA was genotyped for cagA mutation. The IL-8 mRNA expression was measured by Real Time relative quantitation polymerase chain reaction in all Thai tissue samples from the specific site of biopsy, 17 Japanese gastric cancer, and 12 non-cancer gastric mucosal samples. The multivariate analysis was used for the risk study. The student t-test and Mann-Whitney were used for quantitative analysis. The correlation study was done in the subgroup analysis. The STATA 11.0, USA, and SPSS version 16, USA were used for statistical analysis. The p-value 10 = 2.62) and 1509.11 (log 10 =2.17), respectively, p= 0.014. The total mean IL-8 mRNA expression in non-cancer Thais is 2,262 (log 10 =1.49) while that in Japanese non-cancer is 10.79(log 10 = 0.69), p 10 >2 in both Thais and Japanese, Odds ratio= 7.97 (95%CI=3.75-16.97, p 10 ≥2 are significantly cut off risk difference between Thai cancer and non-cancer, p= 0.03 and p Citation Format: Sirikan Yamada, Shunji Kato, Takeshi Matsuhisa, Luksana Makonkawkeyoon, Bandhuphat Chakrabandhu, Thiraphat Chakrabandhu, Takeshi Azuma. The factual evidence of gastric mucosal IL-8 mRNA expression, cagA gene H.pylori infection, and pepsinogen I/II ratio in Thai gastric cancer . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1148. doi:10.1158/1538-7445.AM2013-1148