An international comparability study on quantification of mRNA gene expression ratios: CCQM-P103.1.

Measurement of RNA can be used to study and monitor a range of infectious and non-communicable diseases, with profiling of multiple gene expression mRNA transcripts being increasingly applied to cancer stratification and prognosis. An international comparison study (Consultative Committee for Amount of Substance (CCQM)-P103.1) was performed in order to evaluate the comparability of measurements of RNA copy number ratio for multiple gene targets between two samples. Six exogenous synthetic targets comprising of External RNA Control Consortium (ERCC) standards were measured alongside transcripts for three endogenous gene targets present in the background of human cell line RNA. The study was carried out under the auspices of the Nucleic Acids (formerly Bioanalysis) Working Group of the CCQM. It was coordinated by LGC (United Kingdom) with the support of National Institute of Standards and Technology (USA) and results were submitted from thirteen National Metrology Institutes and Designated Institutes. The majority of laboratories performed RNA measurements using RT-qPCR, with datasets also being submitted by two laboratories based on reverse transcription digital polymerase chain reaction and one laboratory using a next-generation sequencing method. In RT-qPCR analysis, the RNA copy number ratios between the two samples were quantified using either a standard curve or a relative quantification approach. In general, good agreement was observed between the reported results of ERCC RNA copy number ratio measurements. Measurements of the RNA copy number ratios for endogenous genes between the two samples were also consistent between the majority of laboratories. Some differences in the reported values and confidence intervals ('measurement uncertainties') were noted which may be attributable to choice of measurement method or quantification approach. This highlights the need for standardised practices for the calculation of fold change ratios and uncertainties in the area of gene expression profiling.

Effect of oral anabolic steroid on muscle strength and muscle growth in hemodialysis patients.

BACKGROUND AND OBJECTIVES: Sarcopenia is common in hemodialysis patients. This study examined whether the anabolic steroid oxymetholone improves muscle mass and handgrip strength in hemodialysis patients and possible mechanisms that might engender such changes. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Forty-three eligible hemodialysis patients were randomly assigned to ingest oxymetholone or placebo for 24 weeks. Body composition, handgrip strength, and quality of life were measured during the study. Muscle biopsies were performed and analyzed for mRNA levels for myostatin, IGF-I, IGF binding proteins, and myosin heavy chains and protein expression. Muscle fiber types and diameter were assessed by reduced nicotinamide-adenine dinucleotide staining. RESULTS: There was a significantly greater increase in fat-free mass and handgrip strength and decrease in fat mass in the oxymetholone compared with the placebo group. Moreover, compared with baseline values, patients given oxymetholone exhibited an increase in fat-free mass, handgrip strength, physical functioning scores, and type I muscle fiber cross-sectional area and a decrease in fat mass, whereas patients receiving placebo did not undergo changes. There was a significantly greater increase in muscle mRNA levels for myosin heavy chain 2×, IGF-I, and IGF-II receptor with oxymetholone treatment than placebo. Liver enzyme rose significantly in the oxymetholone group, but the number of values greater than three times the upper limit of normal were not different between these groups. CONCLUSIONS: In hemodialysis patients, ingesting oxymetholone was associated with an increase in fat-free mass, handgrip strength, and muscle mRNA levels for several growth factors and a decrease in fat mass, but it also induced liver injury.

Serum zinc, selenium and copper in priest subjects.

OBJECTIVE: To study the serum zinc, selenium, and copper of priest subject who attended the outpatient Department, Priest Hospital Bangkok for a physical check-up, between March and October 2003 MATERIAL AND METHOD: The serum zinc, selenium, and copper of 112 priest subjects, 33 healthy and 79 diseased, compared with a control group of 90 males and 119females Thai volunteers. RESULTS: Serum copper was statistically significantly higher in the priest subjects than in the controls. However, serum selenium in the priest group was significantly lower than in the control group. Higher serum copper, zinc, and selenium levels were show in the male priest group than in the male control group. Of the priests subjects 50.9% (56/110) and 16.5% (18/109) had decreased zinc and selenium levels respectively. The controls had decreased zinc level at 53.4% (39/73) in males, and 34.3% (36/105) in females, and decreased selenium level at 18.6% (35/188). Serum copper concentrations were greater than 140 mg/dl in 44.5% of priest and 30% control subjects. CONCLUSION: Statistically significant difference in serum selenium and copper found in the priest subject when compared with the control subject.

Exposure to lead of boatyard workers in southern Thailand.

Lead oxide is used extensively in the construction and repair of wooden boats in Thailand, but the behaviors of boatyard workers that could place them at risk of contamination have not previously been documented. Baseline data on practices and behaviors of boatyard workers and on the level of worker and workplace contamination with lead were therefore collected. Fifty workers in two boatyards participated in this study. Lead exposure of workers was assessed by determining airborne and blood lead levels. A questionnaire was administered to collect information on work history, suspected exogenous lead sources, personal behavior and knowledge about lead. Evidence obtained by the study indicated that safety behavior and personal hygiene were poor--workers used no mask, gloves or hood, wore open sandals, smoked, drank, chewed and ate during work and did not wash their hands before drinking or eating. Some workers had lunch in the working area. The mean personal airborne lead of caulkers (36.4 microg/m3) was higher than that of carpenters (8.3 microg/m3). Forty-eight percent of all workers and 67% of caulkers had a blood lead level (BLL) exceeding 40 microg/dl. Multiple linear regression indicated that blood lead levels of workers were significantly related to job and education level, with significant differences between boatyards. In addition, the potential for "take-home" contamination was high; none of the workers took a shower or changed their clothes prior to going home. These results indicate a problem of lead exposure of sufficient magnitude to be a public health concern.

HLa-class II (DRB & DQB1) in Thai sudden unexplained death syndrome (Thai SUDS) families (Lai-Tai families).

Thai Sudden Unexplained Death Syndrome (Thai SUDS), or Lai-Tai, is a major health problem among rural residents of northeastern Thailand. The cause has been identified as a genetic disease. SUDS, a disorder found in Southeast Asia, is characterized by an abnormal electrocardiogram with ST-segment elevation in leads V1-V3, identical to that seen in Brugada Syndrome (Brugada Sign, BS) and sudden death due to ventricular fibrillation and cardiac arrest (represents an arrhythmogenic marker that identifies high-risk for SUDS). SUDS victims have a sleeping disorder (narcolepsy). The HLA-DR locus is tightly associated with narcoleptic Japanese patients and HLA-DR2, DQ haplotypes were also found in Oriental narcoleptic patients. These circumstances prompted us to study the association between the disease and HLA Class II by HLA DNA typing using a PCR-SSO method, with five Thai SUDS families (18 BS-positive subjects as the cases, and 27 BS-negatives as the controls). We found that the HLA-DRB1 *12021 allele was significantly increased in BS-positive subjects (p = 0.02; OR = 4.5), the same as the HLA-DRB1*12021-DQB1 *0301/09 haplotype (p = 0.01; OR = 7.95). Our data suggests that the HLA-DRB1* 12021 allele associated with BS and the HLA-DRB1*12021-DQB1 *0301/09 is a haplotype susceptible to arrhythmogenic markers that can identify a high risk for SUDS.

Relationship between serum antioxidant vitamins A, E, and C and lipid profiles in priest subjects at the Priest Hospital.

The serum vitamins A, E, and C (antioxidant vitamins) of 112 priest subjects, compared with 90 males and 119 females in a control group, were investigated. Subjects for the study were Thai volunteers who attended the Outpatient Department, Priest Hospital, Bangkok, for a physical check-up from July to September 2003. There was no age difference between the priest group and the controls. All serum vitamins, A, E, and C, of the priest group were significantly lower than the control group. Statistically significantly higher levels of total cholesterol, LDL-C, and LDL-C/HDL-C ratio were found in the priest subjects compared with the controls. The median serum retinol concentration in the priest subjects was 3.02 micromol/l (range 1.47-4.01 micromol/l) compared with 3.23 micromol/l (range 1.74-4.57 micromol/l) in the controls (p<0.01). The median serum a-tocopherol concentration in the priest subjects was 18.1 mmol/l (range 5.8-27.3 micromol/l) compared with 19.6 mmol/l (range 7.3-37.7 micromol/l) in the controls (p<0.01). The median serum ascorbic acid concentration in the priest subjects was 3.74 mg/l (range 0.0-17.0 mg/l) compared with 6.37 mg/l (range 0.0-18.0 mg/l) in the controls. The median values for retinol, alpha-tocopherol, and ascorbic acid serum concentrations in the male priests were lower than the control males. A total of 28% and 65% of the priest subjects had decreased alpha-tocopherol and ascorbic acid levels, while the controls had decreased alpha-tocopherol and ascorbic acid levels of 20% and 31.5%, respectively. A total of 67.8% and 54.4% of priest and control subjects, respectively, had cholesterol concentrations of > or = 5.18 mmol/l. However, a prevalence of low HDL-C (HDL-C < or = 0.91 micromol/l) was found in 1.8% of priest subjects and 1.4% of controls. Statistically significant associations were found between alpha-tocopherol, cholesterol, LDL-C, triglyceride, and serum retinol. A positive correlation was found between age, retinol, and serum alpha-tocopherol. A negative correlation was found between cholesterol, HDL-C, LDL-C, and the serum alpha-tocopherol/cholesterol ratio. In addition, negative correlations were found between weight, cholesterol, LDL-C, triglyceride, and the serum alpha-tocopherol/(cholesterol + triglyceride) ratio in priest and control subjects. The results suggest more research should be conducted into the health and nutritional problems of both healthy and diseased priest subjects concerning vitamins and oxidative stress.

Relationship of tobacco smoking with serum vitamin B12, folic acid and haematological indices in healthy adults.

OBJECTIVES: To investigate the effects of tobacco smoking on serum vitamin B12, folic acid and haematological parameters in healthy Thai smokers and non-smokers. DESIGN: Cross-sectional study of smokers and non-smokers in a military unit in Bangkok, Thailand. SETTING: A military unit in Thailand. SUBJECTS: One hundred and twenty-three male smokers from a military unit in Bangkok, who participated voluntarily in the study, were investigated. Sixty-six male non-smokers from the same unit were selected as controls. Fasting blood samples were collected for investigation of vitamin B12, folic acid and haematological variables. RESULTS: The serum folic acid concentration of smokers was lower than that of non-smokers, but was not statistically significantly different. Haemoglobin was lower in smokers than in non-smokers; 16.3% of smokers were anaemic compared with only 3.0% of non-smokers. Anaemia was not related to folate deficiency. The white blood cell count was found to be higher in smokers than in non-smokers. CONCLUSION: The results of this study suggest that there were low serum folic acid concentrations in smokers compared with non-smokers, which might contribute to the development of vascular and cardiovascular diseases. The higher white blood cell count might be indicative alterations in the immune functions of smokers.

Serum copper, zinc, ceruloplasmin and superoxide dismutase in Thai overweight and obese.

The serum copper, selenium, ceruloplasmin, superoxide dismutase (SOD) (specific activities of antioxidant enzymes), anthropometric measurements, including waist/hip ratio 51 male and 190 female overweight subjects (body mass index (BMI) > or = 25.0 kg/m2) compared with a 26 male and 83 female control group (BMI = 18.5-24.9 kg/m2) Thai volunteers who attended the Out-patient Department, General Practice Section, Rajvithi Hospital, Bangkok, for a physical check-up from March to October, 1998, were investigated. There was no age difference between the overweight group and the controls. All of the anthropometric variables, except the height of the overweight group, were significantly higher than those of the normal subjects. The medians of weight and waist/hip ratio of overweight and obese males were significantly higher than those of overweight and obese females. Serum ceruloplasmin, copper were statistically significantly higher in overweight subjects than in the controls. However, serum zinc and superoxide dismutase activity in the overweight group were found to be lower than in the control group. Higher serum ceruloplasmin, copper, zinc and superoxide dismutase activity were shown in the female overweight group than in the male overweight group. Ceruloplasmin was found to correlate positively with copper concentration but negatively related with superoxide dismutase enzyme activity. A negative correlation was found between serum copper and zinc concentrations in both sexes of the overweight and obese subjects. Low SOD activity found in the overweight and obese subjects might be caused by low zinc intake.

The relationships between anthropometric measurements, serum vitamin A and E concentrations and lipid profiles in overweight and obese subjects.

The weight, height, body mass index (BMI), waist/hip ratio, serum retinol and alpha-tocopherol and lipid profiles of 16 overweight (BMI > or = 25.0 kg/m2) Thai males and 56 overweight females, compared with 14 males and 58 females in a control group (BMI 18.5-24.9 kg/m2), were investigated. Subjects for the study were those persons who turned up regularly for physical check-up at the Outpatient Department, General Practice Section of Rajvithi Hospital, Bangkok. The study was conducted between December 2000-March 2001. Higher levels of cholesterol, LDL-C, LDL-C/HDL-C ratio were found in the overweight compared with the control subjects. Statistically significant higher triglyceride levels were found in the overweight compared with the control subjects. The median serum retinol concentration in overweight subjects was 2.80 mumol/L (range 0.53-4.62 mumol/L) compared with 2.97 mumol/L (range 1.21-4.12 mumol/L) in control subjects (p = 0.0736). The median serum alpha-tocopherol concentration in overweight subjects was 17.30 mumol/L (range 6.29-28.65 mumol/L) compared with 18.75 mumol/L (range 5.30-30.28 mumol/L) in control subjects (P < 0.05). The median values of retinol and alpha-tocopherol serum concentrations in the overweight and obese males were lower than those of the overweight and obese females. A total of 6.3% (1 out of 16) and 12.5% (2 out of 16) of the overweight/obese males had decreased retinol and alpha-tocopherol levels, while the overweight/obese females had decreased retinol and alpha-tocopherol level of 1.8% (1 out of 56) and 10.7% (6 out of 56), respectively. A total of 12.5% and 39.3% of the overweight/obese males and females had cholesterol concentrations of > or = 6.48 mmol/l. However, the prevalence of low HDL-C (HDL-C < or = 0.91 mmol/l) was found to be 50% in the overweight and obese males and 10.7% in the overweight and obese females. Statistically significant associations were found between age, cholesterol, LDL-C, and serum alpha-tocopherol in the overweight and obese male and female subjects. A negative correlation was found between weight, BMI, AC, MAMC, hip circumference and serum retinol in both the overweight and obese subjects. A negative correlation was found between weight, BMI, MAMC, waist, hip circumferences and serum alpha-tocopherol in both the overweight and obese subjects.

Serum homocysteine, B12 and folic acid concentration in Thai overweight and obese subjects.

This study investigated levels of serum homocysteine, vitamin B12, folic acid, vitamin B6 and vitamin C, in 37 male and 112 female overweight and obese Thai volunteers (body mass index; BMI > or = 25.00), and 23 male and 90 female normal-weight Thai volunteers, who came for a physical check-up at the Out-patient Department, General Practice Section, Rajvithi Hospital, Bangkok from March to October of 2000. Data included anthropometric measurements and waist/hip ratios. All anthropometric variables, except height, were significantly higher for the overweight subjects than for the normal subjects. Statistically significantly higher levels of serum homocysteine were found in the overweight subjects. Serum homocysteine concentrations in overweight and obese males were significantly higher than in overweight and obese females. Serum folic acid and vitamin C in the overweight and obese were found to be statistically significantly lower than in the control subjects. No statistically significant difference in vitamin B12 was found in the overweight and obese subjects compared with the normal control subjects. The medians of serum folic acid and vitamin C concentrations for the overweight and obese males were significantly lower than those of the overweight and obese females. A negative correlation was found between serum folic acid and homocysteine concentrations in all overweight and obese subjects. A significant negative correlation between serum folic acid and vitamin B6 was observed in both male and female overweight and obese subjects. The results of the investigation suggest that homocysteine levels in overweight and obese subjects seem to be caused by insufficient dietary folic acid intake and probably not by B12 deficiency.