DNA extraction and amplification of 10-day, room-temperature blood samples.

DNA was serially studied in 20 samples of buffy coat stored at room temperature. Each sample was divided into 5 equal volumes, namely D0, D3, D5, D7 and D10. DNA extraction was performed on days 0, 3, 5, 7 and 10 after blood collection. The mean ratio of OD260/OD280 of the DNA obtained from D0 to D10 ranged from 1.77 to 1.79, and the mean amounts of the DNA obtained from D0 to D10 ranged from 602 to 740 ng/ul. There were no significant differences in the mean ratio and amounts of DNA obtained among these samples (p > 0.05). Subsequently, amplification was successfully performed from this template DNA to yield products of 1.4 kb and 142 bp at the sites associated with beta globin and factor VIII genes, respectively. These findings suggest the possibility of sending blood samples for DNA analysis by mail, or no ice is required during transportation.

Serum copper and zinc levels in Thai patients with various diseases.

This study was conducted to evaluate the possible alteration of serum Cu and Zn levels in 118 medical inpatients (53 women and 65 men) in Ramathibodi Hospital. Patients were classified according to their main clinical diseases: pulmonary (n = 12), renal (14), infectious (30), malignant (9), cardiovascular (22), GI & hepatic (13) and hematological (18) diseases. Significantly increased serum Cu concentrations were found in patients with pulmonary, malignant, cardiovascular and infectious diseases; moreover, 75, 75, 50 and 37 per cent of these diseases, respectively, had serum levels greater than the normal mean + 2SD (23.6 mumol/L). Besides, 5 per cent of patients (3 in renal, 1 in infectious and 2 in GI & hepatic diseases) had low serum Cu levels suggestive of Cu depletion. By contrast, significantly decreased serum Zn concentrations were found in patients with GI & hepatic, infectious, renal, cardiovascular and malignant diseases. Serum Zn levels below the normal mean - 2SD (8.1 mumol/L) were presented in 46, 37, 29, 23 and 22 per cent of cases, respectively. It was found that serum Cu/Zn ratio in our patients not only with cancer but also with other diseases were statistically significant from the normal group. Hence, the use of serum Cu/Zn ratios as markers for the diagnosis of cancer or for staging tumors must be interpreted cautiously.

Thiamin and riboflavin status of medical inpatients.

Thiamin status was assessed by erythrocyte transketolase activity (ETKA) and thiamin pyrophosphate effect (TPPE) and riboflavin status by erythrocyte glutathione reductase activity (EGRA) and activity coefficient (AC) in 165 medical inpatients in Ramathibodi Hospital. Based on TPPE > 15 per cent, 9 per cent of the medical inpatients had thiamin depletion. Most of them were patients with renal, cardiovascular, hematological and infectious diseases. Based on AC > or = 1.2, 17 per cent of these inpatients had riboflavin depletion. Most of them were patients with pulmonary, cardiovascular and hematological diseases. Only one patient with pulmonary disease had both thiamin and riboflavin depletion. The proportion of thiamin depletion (2/37) in subjects with thiamin supplementation (mean 32.4, median 6, mode 2 md/d) tended to be less than those without (9/98). But, subjects with riboflavin supplementation (mean 3.3, median 4, mode 1 md/d) had the proportion of riboflavin depletion (0/31) significantly (Z-test, p < 0.005) lower than without supplementation (23/104). The data suggested that although the usual dose of vitamin supplementation in medical inpatients is beneficial thiamin depletion can still be present in catabolic patients.

Levels of serum interleukin-6 and tumor necrosis factor in postsplenectomized thalassemic patients.

Levels of serum interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) were studied in 34 nonsplenectomized thalassemic patients (Thal/nonsplenec), 43 postsplenectomized thalassemic patients (Thal/postsplenec), 13 splenectomized non-thalassemic patients (nonThal/ postsplenec) and 18 normal control by enzyme linked immunosorbent assay method. Serum IL-6 concentration in Thal/postsplenec was significantly increased when compared with Thal/ nonsplenec and normal volunteers (3.55 +/- 2.47 pg/ml vs 2.38 +/- 2.31 pg/ml, p = 0.036 and 3.55 +/- 2.47 pg/ml vs 2.66 +/- 0.45 pg/ml, p = 0.028, respectively). This study also demonstrated that TNF-alpha value in Thal/postsplenec was drastically increased above normal control level (15.8 +/- 4.86 pg/ml vs 9.16 +/- 2.18 pg/ml, p = 0.001) and the level was statistically significantly higher than that in Thal/ nonsplenec (15.5 +/- 4.86 pg/ml vs 9.96 +/- 5.19 pg/ml, p = 0.001). There was a trend toward increasing of cytokine levels in Thal/postsplenec with higher platelet count although no correlation was observed. This study addresses the possible role of IL-6 and TNF-alpha in the pathogenesis of reactive thrombocytosis in Thal/postsplenec.