Cytotoxicity of Cratoxylum Formosum Subsp. Pruniflorum Gogel Extracts in Oral Cancer Cell Lines.

BACKGROUND: Oral cancer is a health problem in Thailand. Cratoxylum formosum subsp. pruniflorum Gogel (Teawdang), normally consumed in northeast Thailand, has proven cytotoxic to cervical cancer cell lines including HeLa, SiHa and C-33A. Recently, Asian oral cancer cell lines, ORL-48 and ORL-136, were established. Therefore, we aimed to study cytotoxicity of Teawdang in these. Total phenolic (TPC) and flavonoid content (TFC), and antioxidant activity of Teawdang were also determined. MATERIALS AND METHODS: Teawdang was purchased from Khon Kaen market during June-October 2013. Hexane (CHE), ethyl acetate (CEE) and methanol (CME) extracts of its edible part were analyzed for TPC by the folin-ciocalteau method and for TFC by an aluminium colorimetric method. Antioxidant activity and cytotoxicity in normal Vero cells and oral cancer cells were investigated. Cell viability was assessed using 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assays. RESULTS: CME and CEE had higher TPC and TFC and antioxidant activity than CHE. Both CME and CEE, at 200 µg dry wt/mL, were cytotoxic to the studied oral cancer cell lines. However, CME was cytotoxic to Vero cells whereas CEE was not. Compared to Vero cells, CEE significantly inhibited ORL-48 and ORL-136 growth (p=0.03 and p=0.02, respectively). CONCLUSIONS: CEE exhibited cytotoxic effects on the studied oral cancer cell lines but not normal Vero cells. The bioactive compounds in CEE should be further purified and elucidated for their mechanisms of action for development as anticancer agents.

Anticancer Potential of Cratoxylum formosum Subsp. Pruniflorum (Kurz.) Gogel Extracts Against Cervical Cancer Cell Lines.

BACKGROUND: Most northeast Thai vegetables may play roles in human health by acting as antioxidant and anticancer agents. Recent study showed that Cratoxylum formosum subsp. pruniflorum (Kurz.) Gogel. (Teawdang) could inhibit growth of liver cancer cell lines. Cervical cancer, which has human papilloma virus as its main cause, is found at high incidence in Thailand. Due to increasing drug resistance, searches for potential anticancer compounds from natural source are required. Therefore, our purpose was to evaluate the cytotoxicity of Teawdang extracts in cervical cancer cell lines. MATERIALS AND METHODS: Teawdang edible parts, purchased from Khon Kaen market during July-October 2013 was extracted with organic solvent. Phenolic profiles of crude hexane (CHE), ethyl acetate (CEE), methanol (CME) and water (CWE) extracts were performed by high performance liquid chromatographic (HPLC) techniques. Their cytotoxic effects on cervical cancer cells were investigated with HPV-non infected (C-33A) and HPV-infected (HeLa and SiHa) cell lines. RESULTS: HPLC profiles showed that all crude extracts contained caffeine, ferulic acid and resveratrol. CME and CEE had high contents of gallic acid and quercetin. Catechin was found only in CWE. Cytotoxicity test showed that CEE had the lowest IC50 on HeLa (143.18±13.35 µg/mL) and SiHa cells (106.45±15.73 µg/mL). C-33A cells were inhibited by CWE (IC50 = 130.95±3.83 µg/mL). CONCLUSIONS: There were several phenolic compounds in Teawdang extracts which may have cytotoxic effects on cervical cancer cell lines. Investigation of these bioactive compounds as new sources of anticancer agents is recommended.

Oxidative stress in cancer-bearing dogs assessed by measuring serum malondialdehyde.

BACKGROUND: Oxidative stress, an excess of reactive oxygen species (ROS), causes lipid peroxidation resulting in cell and tissue damages. It may be associated with the development and progression of cancers in dogs. Malondialdehyde (MDA), the end product of lipid peroxidation, is commonly used as a marker of oxidative stress. The objective of this study was to assess oxidative stress in cancer-bearing dogs by measuring serum MDA levels. All client-owned dogs underwent physical examination at the Veterinary Teaching Hospital, Faculty of Veterinary Medicine, Khon Kaen University to determine the health status with the owner's consent. Blood samples of cancer-bearing dogs (N = 80) and clinically normal dogs (N = 101) were obtained and subjected for determination of MDA levels. In addition, complete blood count, creatinine, and alanine aminotransferase were measured. RESULTS: Serum MDA was significantly higher in cancer-bearing dogs than in clinically normal dogs (mean ± SD, 4.68 ± 1.32 µmol/L vs 2.95 ± 0.61 µmol/L, respectively; p < 0.001). Packed cell volume (mean ± SD, 36.18 ± 7.65% vs 44.84 ± 5.54%), hemoglobin (mean ± SD, 11.93 ± 2.88 g% vs 15.17 ± 2.00 g%) and red blood cells (median (IQA), 6.05 (2.15) vs 8.09 (1.34)) were all significantly lower in cancer-bearing dogs than in clinically normal dogs. However, the reverse was true for white blood cells (median (IQA), 18.20 (11.95) vs 14.90 (5.10)). Neither creatinine nor alanine aminotransferase levels were significantly different between groups. CONCLUSIONS: This study supports the conclusion that oxidative stress is associated with many types of cancers in dogs, as serum MDA levels were significantly higher in cancer-bearing dogs compared to clinically normal dogs.

Monosodium glutamate (MSG) intake is associated with the prevalence of metabolic syndrome in a rural Thai population.

BACKGROUND: Epidemiology and animal models suggest that dietary monosodium glutamate (MSG) may contribute to the onset of obesity and the metabolic syndrome. METHODS: Families (n = 324) from a rural area of Thailand were selected and provided MSG as the sole source for the use in meal preparation for 10 days. Three hundred forty-nine subjects aged 35-55 years completed the study and were evaluated for energy and nutrient intake, physical activity, and tobacco smoking. The prevalence of overweight and obesity (BMI ≥ 25 kg/m2), insulin resistance (HOMA-IR >3), and the metabolic syndrome (ATP III criteria) were evaluated according to the daily MSG intake. RESULTS: The prevalence of the metabolic syndrome was significantly higher in the tertile with the highest MSG intake. Further, every 1 g increase in MSG intake significantly increased the risk of having the metabolic syndrome (odds ratio 1.14, 95% confidence interval-CI- 1.12 - 1.28) or being overweight (odds ratio 1.16, 95% CI 1.04 - 1.29), independent of the total energy intake and the level of physical activity. CONCLUSION: Higher amounts of individual MSG consumption are associated with the risk of having the metabolic syndrome and being overweight independent of other major determinants.

Serum antioxidant levels and nutritional status in early and advanced stage lung cancer patients.

OBJECTIVE: Malnutrition frequently occurs in lung cancer patients. We aimed to determine nutritional status and antioxidant and mineral levels in Thai patients with lung cancer. METHODS: A prospective study with matched case-control was conducted. Nutritional status was assessed by body mass index (BMI) and subjective global assessment (SGA). Eastern Cooperative Oncology Group (ECOG) performance status was used to assess the performance. The serum antioxidant and mineral levels were determined. RESULTS: Forty-nine patients with a mean age of 58.8 (range, 35-82) who were first diagnosed with lung cancer were enrolled. They were compared with 60 healthy controls, and levels of retinol, α-tocopherol, ß-carotene, lycopene, ß-cryptoxanthin, selenium, and zinc were lower (P < 0.05). However, peroxidase activity was higher (P = 0.002) in patients. Selenium levels were higher in early stage compared to advanced stage patients (P = 0.041). Overweight patients had higher selenium levels (0.04 mg/L) than normal BMI patients (ß = 0.04, P = 0.035). Patients with SGA class C had lower selenium levels (0.03 mg/L) than those with class A (ß = -0.03, P = 0.035). The poorer ECOG performance patients had significantly lower ß-carotene (ß = -0.192, P = 0.003) and selenium (ß = -0.031, P = 0.011) levels compared with those with good ECOG performance status. CONCLUSIONS: Significantly lower levels of antioxidants and selenium were found in lung cancer patients compared to healthy controls. Levels of some antioxidants and minerals differed among categories of BMI, SGA categories, or ECOG performance status. These findings may be helpful for further studies, such as the effect of nutritional supplementation on clinical outcomes.

Comparative study for the detection of C4d in paraffin-embedded renal allograft biopsies by immunohistochemical techniques.

BACKGROUND: Peritubular capillary complement C4d deposition is one of the criteria for diagnosis of acute humoral rejection in kidney allografts. Little information is provided for the effective technique to stain C4d protein. OBJECTIVE: To compare C4d staining results by using indirect immunofluorescence detection (IF) with immunoperoxidase methods (IP) in formalin-fixed, paraffin-embedded renal allograft tissue. MATERIAL AND METHOD: C4d protein was detected in renal allograft tissues by IF and IP methods. The antigen unmarking procedures were used including (i) heating with 0.05% citraconic anhydride in a water bath plus proteinase K digestion, (ii) heating with 10 mM citrate buffer in a microwave and (iii) digesting with proteinase K for comparing the deposition of C4d in paraffin-embedded tissues. RESULTS: The results showed that the unmarking solution containing 0.05% citraconic anhydride pH 7.4 and heating in a water bath revealed a signal enhancement in the IP method whereas the solution containing 0.05% citraconic anhydride pH 7.4 and heating in a water bath plus proteinase K digestion showed a greatly enhanced signal in the IF method. The prevalence of C4d staining detected in peritubular capillaries was 68% (17/25) and the results observed for both methods were similar. CONCLUSION: 0.05% citraconic anhydride in a water bath with or without proteinase K digestion is useful for unmasking C4d deposition in peritubular capillaries of renal allografts performed by IP and IF methods.

Serum beta-carotene, lycopene and alpha-tocopherol levels of healthy people in northeast Thailand.

Human serum contains many different antioxidants which may be important in the maintenance of antioxidant status. beta-carotene and lycopene are carotenoids with potent antioxidant activity. Carotenoids intake probably protects against cancers and may affect the risk of several chronic conditions. alpha-tocopherol is well known for its function as antioxidant and in reduction of heart disease and cancer risk. We aimed to establish baseline values for serum beta-carotene, lycopene and alpha-tocopherol concentrations in healthy northeast Thais. Fasting serum beta-carotene, lycopene and alpha-tocopherol levels from 294 subjects aged 23-75 years old in northeast Thailand were determined by high performance liquid chromatography (HPLC). The mean serum beta-carotene, lycopene and alpha-tocopherol levels were 0.53 +/- 0.32 micromol/L, 0.57 +/- 0.37 micromol/L, and 26.64 +/- 14.85 micromol/L respectively. Serum beta-carotene and lycopene levels in females (N = 118) were significantly higher than the value for males (N = 176), ie 0.60 +/- 0.31 micromol/L versus 0.48 +/- 0.32 micromol/L (p = 0.002) for beta-carotene and 0.74 +/- 0.38 micromol/L versus 0.46 +/- 0.33 micromol/L (p<0.001) for lycopene whereas alpha-tocopherol level in males (28.60 +/- 14.34 micromol/L) was significantly higher than in females (23.72 +/- 15.16 micromol/L) (p = 0.006). beta-carotene level was positively correlated with alpha-tocopherol (r = 0.22, p<0.001) and lycopene levels (r = 0.63, p<0.001). The results from this study give the baseline data of serum beta-carotene, lycopene and alpha-tocopherol levels in healthy northeast Thai population and also suggest future study on the relationship of dietary intake.

A preliminary study of thiamine status in northeastern Thai children with acute diarrhea.

This study is a preliminary determination of thiamine status in children with diarrhea and metabolic acidosis admitted to hospital. Children with diarrhea (N = 14; age 2 m-6 yr) were divided into 2 groups according to anion gap type; group 1 (21.4%) with a normal anion gap (5.5 +/- 5.2 mmol/l) and group 2 (78.6%) with a wide anion gap (21.2 +/- 5.2 mmol/l). Blood was taken on the day of admission to determine thiamine and lactate levels. Sixty-six point seven percent of patients in group 1 had a normal lactate level (1.5 +/- 0.8 mmol/l) and 33.3% had a high lactate level (2.2 mmol/l); none had thiamine deficiency (TPPE < 20%). High lactate (3.5 +/- 1.4 mmol/l) was found in 54.5% of group 2 and thiamine deficiency was observed in 18.2% of this group. In conclusion, no thiamine deficiency was noted in patients with normal anion gap, but thiamine deficiency was not uncommon in patients with a wide anion gap, regardless of lactic acidosis.

Thiamine deficiency and parasitic infection in rural Thai children.

A study was carried out to determine whether thiamine deficiency in northeast Thailand is linked with parasitic infection. We assessed the thiamine status and tested for parasitic infections in 231 children between 6 and 12 years of age in Nam Phong District of Khon Kaen Province, Thailand. Thiamine deficiency [thiamine pyrophosphate effect (TPPE) > 20%] and parasitic infections (mostly Strongyloides stercoralis) were identified in 16 and 45% of the children, respectively. No association between thiamine deficiency and parasitic infection was found. The general health of the children, according to Thai standard anthropometric curves, was within the normal range. The suboptimal thiamine status and the high prevalence of parasitic infection require attention.

Serum vitamin A and zinc levels of healthy people in northeast Thailand.

BACKGROUND: Vitamin A and zinc are micronutrients which co-related to diseases afflicting northeast Thais. Vitamin A and zinc concentrations in serum have been studied in healthy northeast Thais between 23 and 75 years. METHODS: Vitamin A was analyzed by HPLC and zinc was determined by flame atomic absorption spectrometry. RESULTS: The average serum vitamin A level of the population (n=744) was 2.30 micromol/l (95% CI=2.25-2.35). Males had significantly higher vitamin A levels than females, i.e. 2.61 micromol/l (95% CI=2.53-2.68) vs. 2.03 micromol/l (95% CI=1.98-2.09) (p<0.0001). The vitamin A level of females tended to increase significantly with age (p<0.005), whereas in males levels were relatively constant throughout the age range studied. The average serum zinc level of the population (n=1113) was 18.20 micromol/l (95% CI=18.05-18.36). There was no significant difference in the zinc levels between males and females, i.e. 18.20 micromol/l (95% CI=17.90-18.36) vs. 18.36 micromol/l (95% CI=18.05-18.66). The zinc level tended to decrease significantly as age increased, particularly in the male population (p<0.05). CONCLUSION: The results from this study provide baseline data of serum vitamin A and zinc levels in healthy northeast Thais.