Medical and non-medical correlates of carpal tunnel syndrome in a Taiwan cohort of one million.

BACKGROUND: Carpal tunnel syndrome (CTS), with unclear etiology, is the most common entrapment neuropathy. Its occurrence is related to lots of medical and non-medical conditions with uncertain causality. With a large population, we characterized selected demographical and clinical factors to add more information on CTS-correlated factors and new insight into future CTS prevention. METHODS: A national insurance claim dataset of one million enrollees in Taiwan was used to identify 15 802 patients with CTS and 31 604 randomly selected controls, during a period of 7 years starting 1 January 2000. Statistical association with CTS was determined for five sociodemographic and nine medical factors. RESULTS: Patients were predominantly women (65.6% vs. 47.7% in the control group) and older (40 and above, 62.6% vs. 36.2%). Rheumatoid arthritis was found to be the most significant comorbidity associated with CTS, followed by gout, hypertension, diabetes, obesity, uremia, and acromegaly. For younger group age ≤39, the association of these comorbidities was stronger, and hypothyroidism and vitamin B(6) deficiency were additional comorbidities. Aging appears to reduce the relative impact of the diseases commonly associated with CTS as the possible risk factors. CONCLUSIONS: Identification of the CTS correlates in younger group would be of greater value in timely detection and treatment for these diseases. Correcting these disorders may aid in removing possible causes of CTS. This is the first report on the effect of aging on probable CTS risk factors. How factors associated with aging contribute to the development of CTS remains to be determined.

Toxicological analysis points to a lower tolerable daily intake of melamine in food.

Intensified food safety concern over melamine has prompted national authorities to assess its tolerable daily intake (TDI) for protection of general population including young children. TDI is calculated by dividing a no-observed-adverse-effect level (NOAEL) by a safety factor (SF). Based on appropriate choices of values, the US Food and Drug Administration determined two TDI values in the unit of mg per kg body weight per day as first 0.63 and then 0.063, while the World Health Organization, 0.5 and then 0.2, as a result of increasing the SF values in calculation. We used a similar procedure, with judicious selection of pertinent values, to obtain a TDI of 0.0081. Arguments in support of this lower TDI value were provided to alert the international community.

Toxicity identification evaluation (TIE) of pore water of contaminated marine sediments collected from Hong Kong waters.

Marine sediment can function both as a source and as a sink of marine chemical contaminants. The toxicity of contaminated marine sediment can be assessed by toxic evaluation of its pore water, the inter-particle water of sediment, because toxicants in the pore water may be bioavailable to marine organisms. In this study, the toxicity identification evaluation (TIE) was performed to identify the major toxicants in the pore water of marine sediment collected in Hong Kong waters. In Phase 1 TIE, the suspected toxicants were characterized as anions or organic compounds that are either oxidizable or filterable in alkaline medium. In Phase 2 TIE, the suspected toxicants were identified as sulfide (S(2-)) based on the reduction of toxicity due to lowering of sulfide concentrations by experimental manipulations. The mass balance and spiking analyses in Phase 3 confirmed that S(2-) was one of the major toxicants and that some non-toxic unknown compounds measured by LC-MS, which was removed by C18 solid phase extraction, enhanced the toxicity of S(2-) in the pore water samples.

Application of a toxicity identification evaluation for a sample of effluent discharged from a dyeing factory in Hong Kong.

A first toxicity identification evaluation (TIE) was conducted in three phases using the Microtox test to identify the major toxicant(s) in effluent discharged from a dyeing plant in Hong Kong. In Phase I toxicity characterization indicated that anions were likely to be the major toxicants for the entire effluent. In Phase II concentrations of sulfite and other anions in the original and the anion exchange resin-treated effluent samples were determined by ion chromatography. Anions, which were found in the effluent at comparatively high concentrations and were suspected of being responsible for the toxicity to luminescent bacteria, were selected for further study in Phase III. Investigation in Phase III using the spiking and mass balance approaches confirmed that the sulfite ion was the major toxicant in the effluent.

Dynamics of C2 toxin and chlorophyll-a formation in the dinoflagellate Alexandrium tamarense during large scale cultivation.

The production of paralytic shellfish toxins (PSTs) by the dinoflagellate Alexandrium tamarense ATCI01, a toxigenic strain isolated from South China coastal waters, was studied in batch cultures in relatively large volumes (20l). Under nutrient-replete conditions, this strain produced C2 toxin (C2T) as a predominant PST. In a 15-day production culture, phosphate was depleted by day 4, the stationary phase began at day 6, and the toxin productivity peaked at day 10, in which the cell content of C2T reached 76 fmol per cell. Much of the toxin was produced after the depletion of phosphate in the medium suggesting that C2T is a secondary metabolite. Aeration with small bubbles was useful in increasing cell mass and toxin yield. Chlorophyll-a (Chl-a) was formed in algal cells until the culture entered the stationary phase, after which Chl-a began to disappear rapidly from the culture while the C2T content continued to rise. These results suggest a metabolic relationship between Chl-a and C2T.

Influences of metal concentration in phytoplankton and seawater on metal assimilation and elimination in marine copepods.

Radiotracer experiments were conducted to examine the influence of the concentration of Cd, Se, and Zn in ingested phytoplankton (dinoflagellate Prorocentrum minimum and diatom Thalassiosira weissflogii) and in ambient seawater on metal assimilation and elimination efficiencies of three marine copepods, Acartia spinicauda, Paracalanus aculeatus, and Calanus sinicus. The assimilation efficiencies (AEs) decreased by 1.7 to 2.0 times, 1.4 to 4.1 times, and 1.3 to 2.2 times in the copepods with an increase in metal concentration in ingested algae by 16 to 84 times, 14 times, and 45 to 153 times, for Cd, Se, and Zn, respectively. However, the physiologic turnover rate constant was relatively independent of the metal concentration in copepods. No evidence was found of any interaction between Cd and Zn in their assimilation by copepods. Assimilation efficiencies of Cd were higher in copepods feeding on the dinoflagellate P. minimum, whereas the AEs of Zn were higher in copepods feeding on the diatom T. weissflogii. Differences in metal distribution in algal cytoplasm at different ambient metal concentrations may be partially responsible for the observed influence of metal concentration in algal cells on metal assimilation in copepods. However, metal desorption within the gut of the copepod may have little influence on metal assimilation, as a result of the short gut residence time of food particles and the neutral gut pH. Our study also indicated that the ingestion rate of copepods was reduced by a higher concentration of Cd and Se, but was not affected by Zn concentration in the food particles. Consequently, partial regulation of metal trophic transfer in response to increasing metal contamination may be achieved by a change in metal assimilation efficiency and the ingestion activity of the copepod, but not by changes in metal turnover rates from the animals.

Purification and in vitro folding of recombinant human thrombopoietin receptor expressed in Escherichia coli.

Thrombopoietin receptor (TPOR) is a member of the cytokine receptor superfamily expressed primarily on hematopoietic cells. TPOR plays an important role in regulating platelet production. Due to its low expression level in human tissue, studies on the biochemical and biophysical properties of TPOR have been limited. In the present study, an extracellular domain of recombinant human TPOR (rh TPOR-EN) was expressed in Escherichia coli as inclusion bodies. Purification was achieved by metal chelated chromatography under denaturing condition and was refolded by gel filtration chromatography. Far UV circular dichroism spectroscopy and surface plasmon resonance experiments were performed to demonstrate that the protein was in a refolded state and could bind with its ligand. Thus, a production and purification scheme was developed by which sufficient quantities of rh TPOR-EN can be made available for biochemical and biophysical characterizations.

Cytotoxicity assessment of Ma-huang (Ephedra) under different conditions of preparation.

Ma-huang is a traditional Chinese medicinal herb derived from EPHEDRA: sinica Stapf and other EPHEDRA: species, used to treat asthma, nose and lung congestion, and fever with anhidrosis. It contains 0.5-2.5% by weight of total alkaloids, of which ephedrine accounts for 30 to 90%. Recently, large amounts of ma-huang were used as a source of ephedrine in many dietary supplements formulated for weight reduction, because ephedrine has been found effective in inducing weight loss in diet-restricted obese patients. However, indiscriminate consumption of ma-huang-containing products has resulted in many cases of poisoning, some of which were fatal. The objective of this study is to investigate the relative toxicity of ma-huang extracted under different conditions. The toxicities of various extracts were assayed using MTT colorimetry on a battery of cell lines, while ephedrine alkaloids were analyzed with HPLC. The results are summarized as follows. (1) The cytotoxicity of all ma-huang extracts could not be totally accounted for by their ephedrine contents, suggesting the presence of other toxins in the extracts. (2) Grinding was a significant condition enhancing the toxicity of the extracts. (3) The relatively high sensitivity of the Neuro-2a cell line to the toxicity of ma-huang extracts suggests that the toxic principles were acting on neuronal cells. (4) One condition to produce a ma-huang extract with high ephedrine-to-toxins ratio would be to boil the whole herb for two h.

Determination of interactions between human thrombopoietin and its receptor MPL by yeast two-hybrid system and affinity biosensor.

The binding of human thrombopoietin to the extracellular domain of its receptor MPL prompts a cascade transduction of intracellular signals, leading to the development of megakaryocyte precursors and the production of circulating platelets. We have used a yeast two-hybrid system to reveal, via in vivo interactions between different deletion constructs of MPL and thrombopoietin, that the extracellular subunit 1 of MPL is the ligand binding site and the N-terminal domain of thrombopoietin alone is sufficient for the binding. The extracellular portion of MPL was heterologously expressed in E. coli and its specific affinity with thrombopoietin was visualized in vitro by resonance mirror biosensor technique.

Purification and characterization of the esterases involved in aflatoxin biosynthesis in Aspergillus parasiticus.

The esterases from the cell-free extracts (CFEs) of Aspergillus parasiticus ATCC15517, an aflatoxin-producing strain, catalyzing the hydrolytic conversion of versiconal hemiacetal acetate (VHA) to versiconal was biochemically studied. The specific activity of the enzymes increased 2.5-fold during incubation of mycelia through 40-55 h. No metal ions were required for enzyme stability, but EDTA at 1 mM and dithiothreitol at 0.5-5 mM increased its stability. Three peaks of VHA esterase activity were resolved when the proteins in the CFEs prepared from the mycelia of different ages were separated by anion-exchange column chromatography, suggesting that at least three VHA esterases were present in the eluate of this purification step. One of these esterases extracted from the mycelia of a 55-h culture was partially purified in five steps by means of preparative chromatography and fast protein liquid chromatography. The partially purified enzyme when reacted with [14C]diisopropylfluorophosphate followed by sodium dodecyl sulfate - polyacrylamide gel electrophoresis gave a single radiolabelled band, which corresponded to a protein of 32 kDa. The molecular mass of the partially purified VHA esterase determined with gel filtration was around 60 kDa. The results suggested that the enzyme consists of two isomeric subunits.

Antimutagenicity of ellagic acid against aflatoxin B1 in the Salmonella microsuspension assay.

Ellagic acid (EA) is a phenolic compound with antimutagenic and anticarcinogenic properties. It occurs naturally in some foods such as strawberries, raspberries, grapes, black currants and walnuts. In the present study, we used the Salmonella microsuspension assay to examine the antimutagenicity of EA against the potent mutagen aflatoxin B1 (AFB1) using tester strains TA98 and TA100. Further, we used a two-stage incubation procedure that incorporates washing the bacterial cells free of the incubation mixture after the first incubation to investigate EA and AFB1 interaction. Three different concentrations of AFB1 (2.5, 5 and 10 ng/tube) were tested against five different concentrations of EA for TA98 and TA100. EA significantly inhibited mutagenicity of all doses of AFB1 in both tester strains with the addition of S9. EA alone was not mutagenic at the concentrations tested. The greatest inhibitory effect of EA on AFB1 mutagenicity occurred when EA and AFB1 were incubated together. Lower inhibition was apparent when the cells were first incubated with EA followed by a second incubation with AFB1, and also when the cells were first incubated with AFB1 followed by a second incubation with EA alone. The results of the sequential incubation studies support the hypothesis that one mechanism of inhibition could involve the formation of a chemical complex between EA and AFB1.

Airborne mutagens produced by frying beef, pork and a soy-based food.

Airborne cooking by-products from frying beef (hamburgers), pork (bacon strips) and soybean-based food (tempeh burgers) were collected, extracted, tested for mutagenicity and chemically analysed. The fumes generated by frying pork and beef were mutagenic, with 4900 and 1300 revertants/g of food cooked, respectively. No mutagenicity was detected in fumes from frying tempeh burgers. Bacon fried to a well-done but non-charred state was eight times more mutagenic in a microsuspension Ames/Salmonella test (TA98 with S-9) than hamburgers and about 350 times more mutagenic than tempeh burgers. Among food samples cooked to a well-done, non-charred state, bacon strips had almost 15-fold more mass (109.5 ng/g) than that of the beef, whereas no heterocyclic amine (HCA) was detected in the fried tempeh burgers. 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) was the most abundant HCA, followed by 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) and 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (DiMeIQx). No 2-amino-9H-pyrido[2,3-b]indole (A alpha C) was detected in the food samples fried at about 200 degrees C, although it was present in the collected airborne products. The total amounts of HCAs in the smoke condensates were 3 ng/g from fried bacon, 0.37 ng/g from fried beef and 0.177 ng/g from fried soy-based food. This study indicates that cooks are potentially exposed to relatively high levels of airborne mutagens and carcinogens and that long-term sampling inside restaurants and kitchens may be warranted in order to assess the potential risk of prolonged exposure.

Recent aflatoxin exposure and mutation at codon 249 of the human p53 gene: lack of association.

Experiments were done to show whether a G to T mis-sense mutation at the third base of codon 249 of the p53 tumour suppressor gene is a 'hot spot' of aflatoxin attack as suggested by the results of epidemiological studies. Liver tissue from liver cancer patients in Taiwan and Japan was analysed for the presence of aflatoxin-DNA adducts (ADA) as a marker for aflatoxin exposure and an AGG to AGT transversion at codon 249 of the p53 gene. Ten per cent of samples containing ADA, indicating definite exposure of the subjects to aflatoxin, was found to harbour the codon 249 mutation, whereas 18% of the samples with no detectable adducts also contained the mutation. Our data do not support the hypothesis that codon 249 of the p53 gene DNA is a hot spot for aflatoxin mutagenesis as a 'late stage event' in human hepatocellular carcinogenesis.

Aflatoxin M1 8,9-epoxide: preparation and mutagenic activity.

Treatment of aflatoxin M1 (AFM1) with dimethyldioxirane in an anhydrous mixture of CH2-Cl2 and acetone afforded the corresponding aflatoxin M1 8,9-epoxide (AFM1-E) in practically quantitative yield. This highly reactive intermediate was identified by 1H NMR and characterized by its neat conversion into the corresponding trans-methoxyhydrin derivative 1. The analysis of the 1H NMR spectrum of the above epoxide revealed that one stereoisomer, which should be that with the exo configuration, was present as major component. The mutagenicities of AFM1-E, the parent mycotoxin (AFM1), aflatoxin B1 (AFB1), and its epoxide (AFB1-E) were assessed by using a sensitive improved Ames test with the Salmonella typhimurium strain TA-100. AFM1 and AFB1 had specific mutagenic activities (SMA) of 13 and 121 revertants/ng, respectively, with S9 metabolic activation. AFM1-E was mutagenic with and without metabolic activation showing SMA of 13 and 12 revertants/ng, respectively. AFB1-E had a SMA of 42 and 29 revertants/ng, with and without S9 metabolic enzymes, respectively. These results suggest that the epoxidation of AFM1 can constitute a major route accounting for the cytotoxic effects elicited by this mycotoxin and that AFM1-E is not as active as AFB1-E in reacting with the constituents of the mutagenicity assay.

Comparison of multi-media transport and transformation models: regional fugacity model vs. CalTOX.

Two multimedia environmental transport and transformation computer models are summarized and compared. The regional fugacity model published by Mackay and Paterson (1991), termed Fug3ONT, is a four compartment steady-state model designed to simulate the relative distribution of nonionic organic chemicals in a multimedia system. CalTOX is a seven compartment multimedia total exposure model for hazardous waste sites. Both models are based on the principles of fugacity. CalTOX, however, separates the soil into three layers (surface, root, and vadose) and uses a different approach to estimate the diffusive mass transfer rate in soil. These differences result in lower estimates of the steady-state contaminant concentrations of six environmentally relevant chemicals in the root soil of CalTOX as compared to the bulk soil of Fug3ONT. The difference is greatest for compounds with low mobility in soil such as 2,3,7,8-Tetrachlorodibenzo-p-dioxin and Benzo(a)pyrene where estimates from CalTOX and Fug3ONT differ by more than 3 orders of magnitude. Otherwise, the models provide similar estimates for the distribution of the six chemicals among the air, water, sediment and surface soil.

Aflatoxin toxicity in cultured human epidermal cells: stimulation by 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Aflatoxin B1 (AFB1) at 1 microgram/ml was markedly toxic to human epidermal cells grown in the Rheinwald-Green 3T3 feeder layer system. At 0.1 microgram/ml, the toxicity was barely evident, as assessed by colony expansion during a 2 week exposure, but it was dramatically stimulated by 5 nM 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), which was non-toxic alone. Neither AFB1-dihydrodiol nor AFB2 were toxic in the presence or absence of TCDD, indicating that metabolism to the 8,9-epoxide was responsible for the AFB1 toxicity. Stimulation of AFB1 epoxidation by TCDD was also indicated by the > 20-fold increase in DNA adduct formation in cultures exposed to [14C]AFB1 and TCDD for 4 days as compared to AFB1 alone. Analysis of free metabolites in culture medium by reverse-phase HPLC revealed that confluent epidermal cultures metabolized AFB1 to AFM1, AFB2a and aflatoxicol. In the presence of TCDD, the levels of AFM1 were higher (14 versus 3% of dose) as were those of AFB2a (3 versus 0.5% of dose), while aflatoxicol levels were lower (0.8 versus 2% of dose). In the absence of irradiated 3T3, the toxicities of AFB1, AFB2, AFM1 and aflatoxicol to cells in serum-free medium (0.15 mM Ca2+) were similar to those in the feeder layer system. Although this moderately low calcium concentration appeared quite satisfactory for observing toxicity, the response was attenuated at a lower calcium concentration (0.09 mM Ca2+).

Quantitative integration of the Salmonella microsuspension assay with supercritical fluid extraction of model airborne vapor-phase mutagens.

Vapor-phase mutagens are potentially a major class of toxic contaminants in ambient and indoor air. These compounds are not routinely analyzed due to a lack of an established integrated methodology to quantitatively trap, extract and test the compounds in a bioassay. In a previous report, we emphasized the trapping of volatile and semi-volatile mutagens and the extraction of these compounds using supercritical carbon dioxide (CO2). In the present study, we discuss the use of a bioassay for the quantitation of the model mutagens, ethylene dibromide(EDB) and 4-nitrobiphenyl (4-NB), trapped from an airstream. The compounds EDB and 4-NB were released into a controlled airstream, trapped on XAD-4 adsorbent, and were extracted using supercritical CO2. The extract was tested in a microsuspension modification of the Ames Salmonella/microsome test adapted for volatile compounds. Linear dose-response relationships were obtained for supercritical CO2-extracted EDB using tester strain TA100 (+/- S9) and for 4-NB using tester strains TA98 and TA100 (-S9). Standard dose-response curves with known amounts of the compounds were also determined for comparison with measured amounts of the model compounds collected in an airstream. The gas chromatographic (GC)- and bioassay-determined quantities of EDB and 4-NB were highly correlated, accurate and precise. For example, bioassay-determined EDB concentrations were within 10% of the GC-determined concentrations. Our results demonstrate that the integrated methodology for vapor-phase mutagens developed in this study would be useful for quantitative analysis of these and related airborne vapor-phase mutagenic compounds.

Micronucleus formation in cultured fetal liver blood cells using mitomycin C.

Mouse fetal-liver blood cells were cultured and used to investigate micronucleus formation after exposure to mitomycin C (MMC). The isolated fetal cells were incubated in a medium supplemented with erythropoietin (EPO), and the frequency of micronuclei formation was detected in polychromatic erythrocytes (PCE). The effects of four variables were investigated: (1) MMC exposure dose, (2) MMC exposure time, (3) incubation time, and (4) EPO concentration. PCE were formed by proliferation and differentiation of the erythroid cells in culture. Micronucleated PCE (MNPCE) were observed in a dose-dependent manner after exposing the cultured cells with up to 1.0 microgram/ml MMC. The optimum time of MMC exposure and post-exposure incubation was 3 h and 48 h, respectively, and the optimum EPO concentration was 0.25 U/ml. Mouse fetal-liver PCE are sensitive primordial cell targets that can be obtained in relatively large numbers from a single pregnant animal. The procedure is relatively simple and potentially useful in detecting mutagens and carcinogens capable of causing chromosomal damage.