Evaluation of rapid methods for the determination of okadaic acid in mussels

L.CROCI, A.STACCHINI, L.COZZI, G.CICCAGLIONI, F.MAZZEI, F.BOTRÈ AND L.TOTI. 2001 .
Aims: Two different screening methods, a Buffalo Green Monkey cytotoxicity test and a biosensor test, have been considered to replace the official mouse bioassay in monitoring for okadaic acid (OA) levels in mussels.
Methods and Results: Diarrhoetic shellfish poison-contaminated mussels from the Adriatic Sea were assayed in parallel by means of the mouse bioassay and both alternative methods. Both the cytotoxicity test and the biosensor test showed high sensitivity (OA 0·01 mg g^–1 hepatopancreas and 0·002 mg g^–1 hepatopancreas, respectively) and a high correlation with the mouse bioassay ( r =0·932, P  < 0·001 and r =− 0·850, P  < 0·001, respectively).
Conclusions: Both methods are efficacious, quick, inexpensive and provide data on the amount of toxin present in mussels.
Significance and Impact of the Study: Both methods, besides allowing the simultaneous assay of a great number of samples, comply with the ethical need to reduce the use of animals in the laboratory.     

Development of a F actin-based live-cell fluorimetric microplate assay for diarrhetic shellfish toxins

A new cytotoxicity assay for detection and quantitation of diarrhetic shellfish toxins (DSP) is presented. This assay is based upon fluorimetric determination of F-actin depolymerization induced by okadaic acid (OA)-class compounds in the BE(2)-M17 neuroblastoma cell line. No interferences were observed with other marine toxins such as saxitoxin, domoic acid, or yessotoxin, thus indicating a good specificity of the assay as expected by the direct relationship between protein phosphatase inhibition and cytoskeletal changes. The proposed method is rapid (<2h) and shows a linear response in the range of 50-300 nM OA. The detection limit of the assay for crude methanolic extracts of bivalves lies between 0.2 and 1.0 microg OA per gram of digestive glands, depending on the type of samples (fresh or canned), thus being similar to that of the mouse bioassay. The performance of this assay has been evaluated by comparative analysis of 32 toxic mussel samples by the F-actin assay, mouse bioassay, HPLC and PP2A inhibition assay. Results obtained by the F-actin method showed no differences with HPLC and significant correlation with PP2A inhibition assay (r(2)=0.71). No false negative results were obtained with this new cell assay, which also showed optimum reproducibility.     

Monitoring brevetoxins during a Gymnodinium breve red tide: comparison of sodium channel specific cytotoxicity assay and mouse bioassay for determination of neurotoxic shellfish toxins in shellfish extracts

In October of 1996, a Gymnodinium breve bloom occurred in shellfish harvesting waters of Alabama, Mississippi and Louisiana, Gulf of Mexico, USA. Bloom densities reached 5.6x10(5) cells liter(-1) and bloom residence at shellfish sampling stations ranged from 3 to 28 days. Brevetoxin-2 dominated G. breve toxin profiles in bloom seawater extracts. Shellfish toxicity, assessed by mouse bioassay, exceeded the guidance level for up to 75 days after the bloom had dissipated. Cytotoxicity assays and mouse bioassays showed similar temporal patterns of shellfish toxicity, but the two methods differed in estimations of brevetoxin-3 equivalent toxicity by a factor of 93 to 1. LC-ESI-MS showed the temporal patterns in shellfish toxicity reflected metabolism of G. breve toxins. The molecular ions m/z 1004, 1017 and 1033 dominated LC-ESI-MS spectra of toxic chromatographic fractions from the extracts and were identified as brevetoxin metabolites on the basis of LC-APCI-MS-MS. The discrepancy between cytotoxicity and mouse bioassay estimates of brevetoxin-3 equivalent toxicity resulted from the difference in extraction efficiency of solvents used in the respective methods and the relative sensitivity of the assays to toxin metabolite mixtures present in the extracts. The normalized cytotoxicity assay showed 75% agreement with mouse bioassay positive test samples and 64% agreement with mouse bioassay negative test samples. Published in 1999 by John Wiley & Sons, Ltd.     

Penicillium chrysogenum glucose oxidase -- a study on its antifungal effects

AIMS: Purification and characterization of the high molecular mass Candida albicans-killing protein secreted by Penicillium chrysogenum. METHODS AND RESULTS: The protein was purified by a combination of ultrafiltration, chromatofocusing and gel filtration. Enzymological characteristics [relative molecular mass (M(r)) = 155 000, subunit structure alpha(2) with M(r,alpha) = 76 000, isoelectric point (pI) = 5.4] were determined using SDS-PAGE and 2D-electrophoresis. N-terminal amino acid sequencing and homology search demonstrated that the antifungal protein was the glucose oxidase (GOX) of the fungus. The enzyme was cytotoxic for a series of bacteria, yeasts and filamentous fungi. Vitamin C (1.0 mg ml(-1)) prevented oxidative cell injuries triggered by 0.004 U GOX in Emericella nidulans cultures but bovine liver catalase was ineffective even at a GOX : catalase activity ratio of 0.004 : 200 U. A secondary inhibition of growth in E. nidulans cultures by the oxygen-depleting GOX-catalase system was likely to replace the primary inhibition exerted by H(2)O(2). CONCLUSIONS: Penicillium chrysogenum GOX possesses similar enzymological features to those described earlier for other Penicillium GOXs. Its cytotoxicity was dependent on the inherent antioxidant potential of the test micro-organisms. SIGNIFICANCE AND IMPACT OF THE STUDY: Penicillium chrysogenum GOX may find future applications in glucose biosensor production, the disinfection of medical implants or in the food industry as an antimicrobial and/or preservative agent.     

Decreased flight performance and sperm production in drones of the honey bee (Apis mellifera) slightly infested by Varroa destructor mites during pupal development

We developed a bioassay to measure the flying power of drone, in order to determine which drones could reach a drone congregation area. A wind tunnel was used to test unparasitized drones and drones slightly parasitized by one or two mites during pupal development, and counts were made of the number of spermatozoa that they produced. Drones parasitized with one mite flew as long as control drones (x= 6'55" and 6'48", respectively, P = 0.512); however, those that had been infested by two mites flew significantly less (x= 2'16", P<0.001). There was a significant positive correlation (P<0.01) between flight duration and the number of spermatozoa per drone in control group (r = 0.53), and in both the one mite (r = 0.43) and two mite (r = 0.54) groups. Drones infested during development with one or two mites produced 24 and 45% fewer sperm, respectively.     

Comparison of LST + MUG broth technique and conventional method for the enumeration of Escherichia coli in foods

AIMS: To reduce the analysis time needed for the enumeration of Escherichia coli, a rapid fluorogenic method (MUG) which takes only 48 h was compared with the standard most probable number (MPN) method which takes 6 days as described in the International Standards Organization (ISO). This study provides reliability data for the fluorogenic method applied to certain foods. METHODS AND RESULTS: Both methods were applied to 500 food samples which were analysed for E. coli enumeration. Agreement between the two methods was found in 409 (81 x 8%) samples; 81 (16 x 2%) samples gave higher values by the fluorogenic method, and only 10 (2 x 0%) samples were more effectively assayed by the ISO method. According to statistical analysis, the reliability between the methods was r = 0 x 9706, r(2) = 0 x 9421 and Cronbach's alpha = 0 x 9851. While all three values showed a high degree of correlation (P < 0 x 0001) between the two methods, McNemar's test demonstrated a significant difference between them, indicating that the MUG method was more reliable than the ISO method. CONCLUSIONS: The data suggest that the fluorogenic method is more reliable and shorter to perform than the standard ISO method. SIGNIFICANCE AND IMPACT OF THE STUDY: Comparison of the two methods may provide a rapid and more reliable alternative for the enumeration of E. coli in food samples.     

Inhibition of the reattachment of young adult zebra mussels by single-species biofilms and associated exopolymers

AIMS: To determine the effects of single-species bacterial films and their associated extracellular products on the reattachment of young adult zebra mussels. MATERIALS AND RESULTS: Ten strains of bacteria were isolated from surfaces where adult zebra mussels can be found attached in nature. Single-species biofilms were developed on both glass and polystyrene using these bacteria. The reattachment of zebra mussels (i.e. with byssal threads) was compared between surfaces with and without films. Although no differences were observed in mussel reattachment between glass surfaces with and without films (P > 0.05, anova), a reduction in mussel reattachment between polystyrene surfaces with and without films was observed for seven of the 10 strains (P < or = 0.05 to <0.001, anova). Bacterial extracellular products (BEP) were isolated from five bacterial films and tested for their effects on mussel reattachment. Four of the five sets of isolated extracellular products evoked the same effects as their respective intact biofilms. CONCLUSIONS: We conclude that depending on the substratum, individual strains of bacteria in biofilms can inhibit the reattachment of adult zebra mussels. In some cases, BEP were the source of the inhibitory effects. SIGNIFICANCE AND IMPACT OF THE STUDY: The nature of the substratum on which the biofilms develop affects properties of the biofilm and its extracellular components, which subsequently influences zebra mussel reattachment.     

The relationship between anaerobic performance and muscle metabolic capacity and fibre distribution

Relationships between functional anaerobic indicators and the character of cellular muscle energy metabolism were studied. Twelve untrained male students were tested by a specific anaerobic test on the treadmill. The mean values of the anaerobic test were as follows: blood lactate 10.69 mmol . 1(-1), running speed 16.08 km . h-1 and duration 92.67 s. The average distribution of muscle fibres (m. vastus lateralis) was: type I 52.2%, type II A 29.0% and type II B 18.8%. The mean enzyme activity values were: triosephosphate dehydrogenase (TPDH) 4.67 mu kat . g-1 w.w., lactate dehydrogenase (LDH) 5.76 mu kat . g-1 w.w, citrate synthase (CS) 0.21 mu kat . g-1 w.w. and hydroxyacyl-CoA dehydrogenase (HAD) 0.12 mu kat . g-1 w.w. Significant negative correlations were found between delta LA and CS (r = 0.64) and % of fibre type II B and CS (r = 0.78) and positive correlations between % of fibre type I and CS and/or HAD (r = 0.60 and r = 0.62, respectively).     

Concentration effect of Riesling Icewine juice on yeast performance and wine acidity

AIMS: The objective of this study was to determine the effect of increasing juice soluble solids above 40 degrees Brix on wine yeast's ability to grow and ferment the juice, with particular focus on acetic acid production, titratable acidity (TA) changes and the maximum amount of sugar consumed by the yeast. METHODS AND RESULTS: Riesling Icewine juices at 40, 42, 44 and 46 degrees Brix were inoculated with K1- V116 at 0.5 g 1(-1) and fermented at 17 degrees C until sugar consumption ceased. Increasing soluble solids showed strong negative linear correlations with yeast growth, sugar consumption and ethanol production (r = -0.999, -0.997 and 0.984, P < 0.001, respectively). Acetic acid, glycerol and TA production normalized to sugar consumed showed strong positive correlations to the initial juice concentration (r = 0.992, 0.963, and 0.937, P < 0.001 respectively) but no correlation was found for ethanol production. The acetic acid produced as a function of sugar consumed was positively correlated to the glycerol produced (r = 0.970, P < 0.001). The final TA of the wines ranged between 11.8 and 13.7 g 1(-1) tartaric acid, increasing by 2.3-3 g 1(-1) over the starting juice. The increase in TA was positively correlated to the increase in acetic acid produced after normalizing the data to the amount of sugar consumed (r =0.975, P < 0.001). The acid equivalents resulting from the increase in acetic acid accounted for 80-100% of the TA increase when converted to units of tartaric acid. In the final Icewines, acetic acid represented 19-20% of wine TA. CONCLUSIONS: Increasing Icewine juice concentration from 40 to 46 degrees Brix increases the proportion of yeast sugar metabolism towards glycerol and acetic acid production to cope with the increased osmotic stress by decreasing yeast growth, sugar consumption rate, the total amount of sugar consumed and the total amount of ethanol produced. The high proportional contribution of acetic acid to titratable acidity in Riesling Icewine may affect acidity perception. SIGNIFICANCE AND IMPACT OF THE STUDY: We have determined that 10% v/v ethanol would not be achievable with initial juice concentrations above 42 degrees Brix and that Riesling Icewine juice above 52.5 degrees Brix would be theoretically unfermentable. The high proportional contribution of acetic acid to TA may be an important factor in the organoleptic balance of these Icewines.     

Hematologic iron analyte values as an indicator of hepatic hemosiderosis in Callitrichidae

Hepatic hemosiderosis is one of the most common postmortem findings in captive callitrichid species. Noninvasive evaluation of hematologic iron analytes has been used to diagnose hepatic iron storage disease in humans, lemurs, and bats. This study evaluated the relationship between hematologic iron analyte values (iron, ferritin, total iron binding capacity, and percent transferrin saturation) and hepatic hemosiderosis in callitrichids at the Wildlife Conservation Society's Central Park and Bronx Zoos. Results revealed that both ferritin and percent transferrin saturation levels had strong positive correlations with hepatic iron concentration (P<0.001, r=0.77, n=20; P<0.001, r=0.85, n=10, respectively). Serum iron levels positively correlated with hepatic iron concentration (P=0.06, r=0.56, n=11), but this finding was not significant. Serum total iron binding capacity did not significantly correlate with hepatic iron concentration (P=0.47, r=0.25, n=10). Both ferritin and hepatic iron concentration positively correlated with severity of hepatic iron deposition on histology (P<0.05, r=0.49, n=21; P<0.001, r=0.67, n=21, respectively). This study suggests that ferritin, serum iron concentration, and percent transferrin saturation are convenient, noninvasive, antemortem methods for assessing severity of hemosiderosis in callitrichids.     

The influence of packaging methodology on the microbiological and fatty acid profiles of refrigerated African catfish fillets

AIMS: The shelf-life of refrigerated catfish fillets was determined at 2 degrees C, to simulate retail conditions, using two types of packaging materials, vacuum packing (VP) and oxygen permeable packaging (OPP). METHODS AND RESULTS: Representative samples (n=5) from both types of packaging methods were drawn at random every 2 d until a microbiological count of 106 cfu g-1 was reached. Samples were pooled and screened microbiologically using standard methods. Fatty acid analyses of total lipids, neutral lipid, glycolipid and phospholipid fractions were also conducted, to determine at which point the fish was regarded as spoiled and which packaging method provided a longer shelf-life. OPP limits storage to a maximum of 4 d (aerobic plate count of 8.2 x 105 cfu g-1), whereas VP extends the shelf-life of the fillets to between 6 and 8 d (aerobic plate count of 9.2 x 104 cfu g-1 and 1.66 x 106 cfu g-1, respectively). Similarly, coliform counts increased with time; however, packaging material had no statistical influence thereon. CONCLUSION: Until d 13, when the experiment was terminated, no deterioration in lipid composition of the various fractions was noted. SIGNIFICANCE AND IMPACT OF THE STUDY: An extended shelf-life microbiologically-speaking, for potential processors, could thus be obtained by using VP instead of OPP.     

Apolipoprotein A-V, triglycerides and risk of coronary artery disease: the prospective Epic-Norfolk Population Study

In mouse models, apolipoprotein A-V (apoA-V) exhibits triglyceride (TG)-lowering effects. We investigated the apoA-V/TG relationship and the association of apoA-V with coronary artery disease (CAD) risk by determining serum apoA-V levels and genotypes in a nested case-control (n = 1,034/2,031) study. Both univariate and multivariate apoA-V levels showed no association with future CAD (P = 0.4 and 0.5, respectively). Unexpectedly, there was a significant positive correlation between serum apoA-V and TG in men and women (r = 0.36 and 0.28, respectively, P < 0.001 each) but a negative correlation between apoA-V and LPL mass (r = -0.14 and -0.12 for men and women respectively, P < 0.001 each). The frequency of the c.56C>G polymorphism did not differ between cases and controls despite significant positive association of c.56G with both apoA-V and TG levels. For -1131T>C, the minor allele was significantly associated with lower apoA-V yet higher TG levels and was overrepresented in cases (P = 0.047). The association of -1131T>C with CAD risk, however, was independent of apoA-V levels and likely acts through linkage disequilibrium with APOC3 variants. The positive correlation of apoA-V levels with TG levels, negative correlation with LPL levels, and lack of association with CAD risk highlight the need for further human studies to clarify the role of apoA-V.     

Human chondrocyte senescence and osteoarthritis

Although osteoarthritis (OA) is not an inevitable consequence of aging, a strong association exists between age and increasing incidence of OA. We hypothesized that this association is due to in vivo articular cartilage chondrocyte senescence which causes an age-related decline in the ability of the cells to maintain articular cartilage, that is, increasing age increases the risk of OA because chondrocytes lose their ability to replace their extracellular matrix. To test this hypothesis, we measured senescence markers in human articular cartilage chondrocytes from 27 donors ranging in age from one to 87 years. The markers included expression of the senescence-associated enzyme beta-galactosidase, mitotic activity measured by 3H-thymidine incorporation, and telomere length. beta-galactosidase expression increased with age (r=0.84, p=0.0001) while mitotic activity and mean telomere length declined (r=-0.774, p=0.001 and r=-0.71, p=0.0004, respectively). Decreasing telomere length was strongly correlated with increasing expression of beta-galactosidase and decreasing mitotic activity. These findings help explain the previously reported age related declines in chondrocyte synthetic activity and responsiveness to anabolic growth factors and indicate that in vivo articular cartilage chondrocyte senescence is responsible, at least in part, for the age related increased incidence of OA. The data also imply that people vary in their risk of developing OA because of differences in onset of chondrocyte senescence; and, the success of chondrocyte transplantation procedures performed to restore damaged articular surfaces in older patients could be limited by the inability of older chondrocytes to form new cartilage. New efforts to prevent the development or progression of OA might include strategies that delay the onset of chondrocyte senescence or replace senescent cells.     

Sensitivity of three thyrotropin receptor antibody assays in thyroid-associated orbitopathy

BackgroundThyrotropin receptor autoantibodies (TSH-RAb) are indispensable biomarkers in the laboratory assessment of thyroid-associated orbitopathy (TAO). Clinical sensitivity of three different assays for TSH-R-Ab determination was evaluated in patients with TAO.Methods87 consecutive TAO patients were enrolled and their serum samples analyzed in parallel with three assays. An ECLIA competitive binding and a chemiluminescent bridge immunoassay were used to measure total and binding TSH-R-Ab concentration, while their functional activity was determined using a stimulatory TSH-R-Ab (TSAb) cellbased bioassay.ResultsCompared to the two binding assays (ECLIA p<0.001, bridge p=0.003), the TSAb bioassay was more sensitive pertaining to the positive detection of TSH-R-Ab in TAO patients. No difference (p=0.057) was noted between the ECLIA and bridge assays regarding sensitivity rate. All patients with active and/or moderate-to-severe TAO tested positive in the TSAb bioassay (100% and 100%, respectively), while the positivity rates for bridge and ECLIA binding assays were 89.7% and 82.1% for active TAO, and 90.2% and 86.3% for severe TAO, respectively. Negative predictive values of the bioassay, bridge, and ECLIA assays were 100%, 75%, and 71%, respectively for active TAO, and 100%, 86%, and 71%, respectively for moderate-to-severe TAO. The superiority of the bioassay was most prominent in euthyroid (ET) TAO. Positivity rates of the TSAb bioassay, bridge and ECLIA binding assays were 89.6%, 75%, and 64.6%, respectively for inactive TAO; 86.1%, 69.4%, and 52.8%, respectively for mild TAO; 87.5%, 62.5%, and 12.5%, respectively for euthyroid TAO. The bridge assay correlated better with the ECLIA binding assay (r=0.893, p<0.001), compared to the bioassay (r=0.669, p<0.001).ConclusionsIn patients with TAO of various activity and severity, the TSAb bioassay demonstrates a superior clinical performance compared to both ECLIA and bridge binding assays.     

Separation of iodothyronines from iodinated metabolites in serum from reverse T3 tracer kinetic studies in man and dog: a comparison of two methods

Iodothyronine separation from free iodide and iodoalbumin in serial serum samples obtained from 7 human and 5 dog studies following intravenous injection of radiolabeled reverse triiodothyronine (reverse T3) was compared using acidified ammonium acetate/tetrahydrofuran (THF) elution from C-18 SEP-PAK cartridges or ethyl acetate/butanol (EAB) extraction. Both methods excluded greater than 98% free iodide and greater than 99% iodoalbumin from the iodothyronine fraction. Recovery of labeled reverse T3 was higher for the THF/SEP-PAK (79.4 +/- 4.1%) than for the EAB method (43.2 +/- 6.1%, P less than 0.001), and intra-assay coefficients of variation were lower (2.1 +/- 0.6% and 4.4 +/- 2.0%, respectively, P less than 0.001); HPLC analysis of iodothyronine fractions revealed a single peak co-migrating with injected tracer. The THF/SEP-PAK technique allowed use of larger serum samples at later time points. Serum disappearance curves derived from these two methods were highly correlated in all cases (r = 0.998, P less than 0.001), as were fits of data to sums of exponentials and calculated serum kinetic parameters.     

Detection of bacteria originally isolated from Alexandrium spp. in the midgut diverticula of Mytilus edulis after water-borne exposure

Bacteria associated with toxic dinoflagellates have been implicated in the production of paralytic shellfish poisoning (PSP) toxins, but it has not been substantiated that bacteria are truly capable of autonomous PSP toxin synthesis or what role bacteria may play in shellfish toxification. In this study, different putatively PSP toxin producing bacteria originally isolated from toxic Alexandrium spp. were exposed to the blue mussel Mytilus edulis. To document that these bacteria accumulated in the digestive tract of the mussels, hybridization techniques that use rRNA targeted oligonuceotides for in situ identification of these bacteria were applied. The mussel hepatopancreas was dissected and paraffin and frozen sections were made. The dissected glands were hybridized with digoxigenin-labelled 16S rRNA oligonucleotide probes. Results demonstrate that mussels will readily uptake and accumulate these bacteria in the hepatopancreas. However, the mussels were not rendered toxic by the ingestion of the bacteria as determined by HPLC with UV detection for PSP toxins and determination of sodium channel blocking activity using the mouse neuroblastoma assay. Thus, although the role that bacteria play in mussel toxification remains unclear, methods are now available which will aid in further investigation of this relatively unexplored area.     

Comparison of serum estradiol to urinary estrone conjugates in the rhesus macaque (Macaca mulatta)

Paired urine and serum samples were collected daily during fourteen nonconceptive (7 females) and ten conceptive (9 females) ovarian cycles from a total of 12 female rhesus monkeys (Macaca mulatta). Daily urine samples were analyzed for concentrations of estrone conjugates (Ei Conj). Serum samples were evaluated for concentrations of estradiol (E2) and progesterone (P) by radioimmunoassay (RIA), and bioactive luteinizing hormone (bLH) and monkey chorionic gonadotropin (mCG) were analyzed by a mouse Leydig cell bioassay. Linear correlation (r) between urinary E1 Conj and serum E2 (r range = -0.176-0.948) during nonconceptive cycles aligned by the preovulatory E1 Conj peak (Day 0) improved when daily hormone values were realigned to account for an approximately 24-h delay in the excretion of hormonal metabolites in urine (r range = 0.465-0.967). Similarly, correlation between urinary E1 Conj and serum E2 during conceptive cycles aligned by Day 0 (r range = 0.300-0.824) improved when values were offset by 24 h (r range = 0.408-0.876). When conceptive cycles were compared to nonconceptive cycles, serum P levels were significantly elevated over nonconceptive levels by Day +12 (p less than 0.001), and urinary E1 Conj levels by Day +13 (p less than 0.02), whereas serum E2 and bLH were both significantly elevated by Day +14 (p less than 0.0006 and p less than 0.01, respectively). In both nonconceptive and conceptive cycles, urinary E1 Conj paralleled serum E2 and demonstrated incremental increases above baseline levels, which were greater than for serum E2.     

Predicting rodent carcinogenicity of Ames test false positives by in vivo biochemical parameters

28 chemicals known to be mutagenic in the Ames test but not carconigenic in rodent bioassays were selected for study. The chemicals were administered by gavage in 2 dose levels to female Sprague-Dawley rats. The effects of these 28 chemicals on 4 biochemical assays (hepatic DNA damage by alkaline elution (DD), hepatic ornithine decarboxylase activity (ODC), serum alanine aminotransferase activity (ALT), and hepatic cytochrome P-450 content (P450)) were determined. The scientific approach taken was to either experimentally find individual cancer predictors of high specificity or to mathematically create composite predictors of high specificity.

Composite predictive parameters are defined as follows: CP = [ODC and P450], CT = [ALT and ODC] and TS = [DD or CP or CT]. The specificity (percent of rodent noncarcinogens which test negative) of DD, ODC, ALT, P450, CP, CT and TS was 100%, 46%, 89%, 86%, 93%, 93% and 86%, respectively. For these 28 mutagenic noncarcinogens, the specificity of structural alerts (SA) 13%, mutation in mouse lymphoma cells (MOLY) 0%, chromosomal aberrations in Chinese hamster ovary cells (ABS) 13%, and sister-chromatid exchange in Chinese hamster ovary cells (SCE) 0% were much lower. The k_e test, an experimental measure of electron attachment, had a specificity of 33%. DD was the only DNA related parameter to predict well the noncarcinogenic rodent bioassay result of Ames false-positive chemicals. 5 nongenotoxic parameters (ALT, P450, CP, CT and [CP or CT]) predicted the rodent bioassay result well. Depending on the prevalence of chemicals carcinogenic to humans, the problem of Ames test false positives for predicting human cancer may be either small or large.     

Does acute whole-body vibration training improve the physical performance of people with knee osteoarthritis?

ABSTRACT: Salmon, JR, Roper, JA, and Tillman, MD. Does acute whole-body vibration training improve the physical performance of people with knee osteoarthritis? J Strength Cond Res 26(11): 2983-2989, 2012-The purpose of this study was to determine the effects of a single session of whole-body vibration training (WBVT) on the physical performance of individuals with knee osteoarthritis (OA) in 3 tests designed to simulate activities of daily living (ADLs). Fifteen individuals with symptomatic knee OA completed the Timed-Up-and-Go Test, step test, 20-m walk test, and visual analog scale (VAS) recordings of knee pain intensity. A main effect was detected for time to complete the step test (F[2,28] = 6.243, p = 0.006, (Equation is included in full-text article.)). Post hoc analyses revealed that the time to complete the step test at 5 minutes after WBVT improved significantly (p = 0.042) from that of the pretest. A moderate correlation (r = 0.465, p = 0.001) was found between the VAS scores and the time to complete the step test across all trials. A main effect was found for time to complete the walk test (F[2,28] = 4.370, p = 0.022, (Equation is included in full-text article.)). Post hoc analyses did not indicate significant improvements from pretest seen at 5 minutes after WBVT (p = 0.110) and 1 hour after WBVT (p = 0.224). The WBVT was well tolerated in nearly all the participants, and we observed that an acute bout of WBVT was effective in improving the ability of individuals with knee OA to perform a step test and 20-m walk test. Our findings suggest that WBVT may be an effective nonpharmacologic modality to treat some knee OA symptoms and improve ADLs.     

Effects of temperature and temperature acclimation on survival of zebra mussels (Dreissena polymorpha) and Asian clams (Corbicula fluminea) under extreme hypoxia

Following acclimation to 5°, 15° or 25°C for 14days, samples of 30 Dreissena polymorpha (zebra mussels) andCorbicula fluminea (Asian clams) were held in either aerated(control) or extremely hypoxic N₂ gassed water (PO2 < 3%of full air saturation). Mortality was negligible in all aeratedcontrols. Mean hypoxia tolerance in D. polymorpha ranged from3–4 days at 25°C to 38–42 days at 5°C. Hypoxiatolerance time of zebra mussels increased significantly withdeclining test temperature (P < 0.001) and increasing acclimationtemperature (P < 0.001). Larger zebra mussels were more tolerantthan smaller individuals. Asian clams were 2–7 times moretolerant of hypoxia than zebra mussels, surviving a mean of11.8 and 35.1 days at 25°C and 15°C, respectively, andwithout mortality for 84 days at 5°C, and were not influencedby temperature acclimation. At 25°C, larger specimens ofAsian clams were less tolerant of hypoxia than smaller individuals.Both species are amongst the least hypoxia tolerant freshwaterbivalve molluscs, reflecting their prevalence in well-oxygenatedshallow water habitats. Prolonged exposure to extreme hypoxiamay provide an efficacious control strategy, particularly forD. polymorpha (Received 12 January 1998; accepted 30 September 1998)