Similar scaling of contralateral and ipsilateral cortical responses during graded unimanual force generation.

Hemibody movements are strongly considered as being under the control of the contralateral hemisphere of the cerebral cortex. However, some neuroimaging studies have found a bilateral activation of either the primary sensori-motor (SM1) areas or the rostral prefrontal cortex (PFC), during unimanual tasks. More than just bilateral, the activation of these areas was found to be symmetrical in some studies. However, the symmetrical response remains strongly controversial notably for handgrip force generations. We therefore aimed to examine the bilateral SM1 and rostral PFC area activations in response to graded submaximal force generation during a unilateral handgrip task. Fifteen healthy subjects performed 6 levels of force (ranging from 5 to 50% of MVC) during a handgrip task. We concomitantly measured the activation of bilateral SM1 and rostral PFC areas through near-infrared spectroscopy (NIRS) and the electromyographic (EMG) activity of the bilateral flexor digitorum superficialis (FDS) muscles. Symmetrical activation was found over the SM1 areas for all the investigated levels of force. At the highest level of force (i.e., 50% of MVC), the EMG of the passive FDS increased significantly and the ipsilateral rostral PFC activation was found more intense than the corresponding contralateral rostral PFC activation. We suggest that the visuo-guided control of force levels during a handgrip task requires the cross-talk from ipsi- to contralateral SM1 to cope for the relative complexity of the task, similar to that which occurs during complex sequential finger movement. We also propose alternative explanations for the observed symmetrical SM1 activation including (i) the ipsilateral corticospinal tract and (ii) interhemispheric inhibition (IHI) mechanism. The increase in EMG activity over the passive FDS could be associated with a release of IHI at 50% of MVC. Finally, our results suggest that the greater ipsilateral (right) rostral PFC activation may reflect the greater demand of attention required to control the motor output at high levels of force.

A 12-channel, real-time near-infrared spectroscopy instrument for brain-computer interface applications.

A continuous wave near-infrared spectroscopy (NIRS) instrument for brain-computer interface (BCI) applications is presented. In the literature, experiments have been carried out on subjects with such motor degenerative diseases as amyotrophic lateral sclerosis, which have demonstrated the suitability of NIRS to access intentional functional activity, which could be used in a BCI as a communication aid. Specifically, a real-time, multiple channel NIRS tool is needed to realise access to even a few different mental states, for reasonable baud rates. The 12-channel instrument described here has a spatial resolution of 30 mm, employing a flexible software demodulation scheme. Temporal resolution of approximately 100 ms is maintained since typical topographic imaging is not needed, since we are only interested in exploiting the vascular response for BCI control. A simple experiment demonstrates the ability of the system to report on haemodynamics during single trial mental arithmetic tasks. Multiple trial averaging is not required.

Characterization of the cellulolytic and hydrogen-producing activities of six mesophilic Clostridium species.

AIMS: To characterize cellulolytic, hydrogen-producing clostridia on a comparable basis. METHODS AND RESULTS: H(2) production from cellulose by six mesophilic clostridia was characterized in standardized batch experiments using MN301 cellulose, Avicel and cellobiose. Daily H(2) production, substrate degradation, biomass production and the end-point distribution of soluble fermentation products varied with species and substrates. All species produced a significant amount of H(2) from cellobiose, with Clostridium acetobutylicum achieving the highest H(2) yield of 2.3 mol H(2) mol(-1) hexose, but it did not degrade cellulose. Clostridium cellulolyticum and Clostridium populeti catalysed the highest H(2) production from cellulose, with yields of 1.7 and 1.6 mol H(2 )mol(-1) hexose from MN301 and 1.6 and 1.4 mol H(2) mol(-1) hexose from Avicel, respectively. These species also achieved 25-100% higher H(2) production rates from cellulose than the other species. CONCLUSIONS: These cellulolytic, hydrogen-producing clostridia varied in H(2) production, with Cl. cellulolyticum and Cl. populeti achieving the highest H(2) yields and cellulose degradation. SIGNIFICANCE AND IMPACT OF THE STUDY: The fermentation of cellulosic materials presents a means of H(2) production from renewable resources. This standardized comparison provides a quantitative baseline for improving H(2) production from cellulose through medium and process optimization and metabolic engineering.

Validation of the NFL-225 test for predicting 1-RM bench press performance in college football players.

BACKGROUND: The purpose of this study was to evaluate the accuracy of repetitions-to-fatigue (RTF) using an absolute load of 102.3 kg (225 lbs) to estimate one-repetition maximum (1-RM) bench press performance in college football players using various prediction equations. EXPERIMENTAL DESIGN: a prospective study on the association between muscular endurance and muscular strength. PARTICIPANTS: 260 players from NCAA Division IA (n=43), IAA (n=63), II (n=129), and red-shirts (n=25) were evaluated at the conclusion of a minimum of eight weeks of heavy-resistance training during the off-season. MEASURES: all subjects performed a 1-RM bench press and RTF using an absolute load of 102.3 kg. RESULTS: The Mayhew et al. NFL-225 equation nonsignificantly overestimated 1-RM from RTF by 0.5 kg, while the Chapman et al. NFL-225 equation significantly underpredicted by 3.2 kg, although both equations were comparable in the number of players predicted within +/-4.5 kg of actual 1-RM (52% vs 51%, respectively). Only two of nine RTF equations currently in use produced predicted 1-RM values that were not significantly different from actual 1-RM performance. CONCLUSIONS: Specific NFL-225 equations are more accurate in estimating 1-RM bench press from absolute muscle endurance in college football players than previous published RTF equations. The accuracy of prediction decreases at higher repetitions.

Efficacy and reproducibility of a produce wash in killing Salmonella on the surface of tomatoes assessed with a proposed standard method for produce sanitizers.

The reproducibility of a method developed to evaluate point-of-use sanitizers for fresh produce was tested at three different laboratories. Mixtures of five Salmonella serotypes were inoculated on the surface of ripe tomatoes. After the inoculum was dry, tomatoes were placed inside a plastic bag and sprayed with sterile USP water, Dey and Engley (D/E) neutralizer broth, or a prototype Fit produce wash (PW), an alkaline solution comprised of generally recognized as safe ingredients (water, oleic acid, glycerol, ethanol, potassium hydroxide, sodium bicarbonate, citric acid, and distilled grapefruit oil), and rubbed for 30 s. The tomatoes were rinsed 10 s with 195 ml of D/E neutralizer broth (rinse solution), then combined with 20 ml of D/E neutralizer (residual wash solution) and rubbed by hand to remove residual Salmonella. Populations of Salmonella were determined for each tomato in the rinse solution and residual wash solution. Treatment with PW resulted in reductions in the number of Salmonella 2 to 4 logs greater than those achieved with the sterile water or D/E neutralizer broth controls. Consistent results were obtained across the three study sites, indicating reproducible results were obtained using the test method. The method used to determine the efficacy of killing or removing Salmonella from tomatoes in this study is suggested as a standard method for measuring the efficacy of sanitizers on tomatoes and other similar fruits and vegetables with rigid, smooth surfaces.

Development of a proposed standard method for assessing the efficacy of fresh produce sanitizers.

A series of studies was done for the purpose of developing a proposed standard method to evaluate point-of-use home sanitizers for fresh produce. Preliminary experiments were done to determine the survival of Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes after inoculation onto the surface of ripe tomatoes and drying for up to 24 h at 22 +/- 2 degrees C. Within 2 h, the initial population (6.88 log10 CFU/tomato) of E. coli O157:H7 was reduced by approximately 3 log10, while reductions in similar initial populations of Salmonella and L. monocytogenes were approximately 1 and 0.6 log10 CFU/tomato, respectively, after 40 min and 3 h. A pilot study evaluated treatment with 200 ppm free chlorine and a prototype Fit produce wash (Fit) for their efficacy in killing a five-serotype mixture of Salmonella or L. monocytogenes spot inoculated on tomatoes using the proposed inoculation and recovery procedures. Inoculated tomatoes were sprayed with chlorinated water, Fit, or sterile distilled water (control) and hand rubbed for 30 s. Each tomato was then placed in a plastic bag and rinsed with 200 ml of sterile water by vigorously agitating for 30 s to simulate a procedure consumers might use for sanitizing and rinsing produce in a home setting. Each tomato was transferred to a second bag, and 20 ml of sterile 0.1% peptone was added; tomatoes were rubbed by hand for 40 s. Populations of Salmonella or L. monocytogenes in the rinse water and the 0.1% peptone wash solution were determined. Treatment with 200 ppm chlorine and Fit resulted in > or = 3.07 and > 6.83 log10 reductions, respectively, in Salmonella. Treatment with 200 ppm chlorine and Fit reduced the number of L. monocytogenes by > or = 3.33 and > or = 4.96 log10 CFU/tomato, respectively. The proposed standard method for testing the efficacy of point-of-use produce sanitizers needs to be evaluated for reproducibility of results through a larger scale series of experiments.

Comparison of chlorine and a prototype produce wash product for effectiveness in killing Salmonella and Escherichia coli O157:H7 on alfalfa seeds.

Outbreaks of Salmonella and Escherichia coli O157:H7 infections associated with alfalfa and other seed sprouts have occurred with increased frequency in recent years. This study was undertaken to determine the efficacy of a liquid prototype produce wash product (Fit), compared with water and chlorinated water, in killing Salmonella and E. coli O157:H7 inoculated onto alfalfa seeds. We investigated the efficacy of treatments as influenced by seeds from two different lots obtained from two seeds suppliers and by two methods of inoculation. The efficacy of treatments was influenced by differences in seed lots and amount of organic material in the inoculum. Significant (alpha = 0.05) reductions in Salmonella populations on seeds treated with 20,000 ppm of chlorine or Fit for 30 min ranged from 2.3 to 2.5 log10 CFU/g and 1.7 to 2.3 log10 CFU/g, respectively. Reductions (alpha = 0.05) in E. coli O157:H7 ranged from 2.0 to 2.1 log10 CFU/g and 1.7 to more than 5.4 log10 CFU/g of seeds treated, respectively, with 20,000 ppm of chlorine or Fit. Compared with treatment with 200 ppm of chlorine, treatment with either 20,000 ppm of chlorine or Fit resulted in significantly higher reductions in populations of Salmonella and E. coli O157:H7. None of the treatments eliminated these pathogens as evidenced by their detection on enrichment of treated seeds. Considering the human health and environmental hazards associated with the use of 20,000 ppm of chlorine, Fit provides an effective alternative to chlorine as a treatment to significantly reduce bacterial pathogens that have been associated with alfalfa seeds.

The Bruce treadmill protocol: does walking or running during the fourth stage alter oxygen consumption values?

OBJECTIVE: To investigate heart rate (HR) and relative oxygen consumption (VO2) measures during two modes (walking or running stages four) of the Bruce treadmill protocol. PARTICIPANTS: Male volunteers (n = 27), ranging in age from 25 to 56 years (M = 39.1 +/- 10.7 yrs). EXPERIMENTAL DESIGN: S's performed to volitional fatigue on the two randomly assigned treadmill tests. MEASURES: HR and VO2 were taken each minute and at point of exhaustion. RESULTS: Dependent "t"-tests revealed a significantly (p < or = 0.05) difference between the protocols at 11 minutes (running = 46.7 +/- 3.9 > walking = 44.6 +/- 3.7 ml[kg.min-1]) and at 12 minutes (running = 49.3 +/- 4.1 > walking = 47.6 +/- 3.5 ml[kg.min-1]) on the VO2 values. A significant differences was noted on HR at 11 minutes (running = 158.1 +/- 13.5 > walking = 156.0 +/- 13.0 bpm) and at 12 minutes (running = 160.4 +/- 11.0 > walking = 157.8 +/- 11.4 bpm) between the protocols. The two-way ANOVA technique revealed no significant differences or interactions on VO2 or HR between younger (< 45 yrs) and older (> or = 45 yrs) subjects during either protocol. A one-way ANCOVA indicated no significant differences between taller and shorter subjects on VO2 during the fourth stage of the Bruce protocol. The correlations, between the two protocols, for HR were strong but were weaker and inconsistent for VO2. The repeated measures ANOVA indicated significant within subject variability between test administrations. CONCLUSIONS: When testing endurance trained males, modality, age and height are not factors in differences of VO2 values during the 4th stage of the Bruce treadmill test but learning effect could be.

Direct 5S rRNA Assay for Monitoring Mixed-Culture Bioprocesses.

This study demonstrates the efficacy of a direct 5S rRNA assay for the characterization of mixed microbial populations by using as an example the bacteria associated with acidic mining environments. The direct 5S rRNA assay described herein represents a nonselective, direct molecular method for monitoring and characterizing the predominant, metabolically active members of a microbial population. The foundation of the assay is high-resolution denaturing gradient gel electrophoresis (DGGE), which is used to separate 5S rRNA species extracted from collected biomass. Separation is based on the unique migration behavior of each 5S rRNA species during electrophoresis in denaturing gradient gels. With mixtures of RNA extracted from laboratory cultures, the upper practical limit for detection in the current experimental system has been estimated to be greater than 15 different species. With this method, the resolution was demonstrated to be effective at least to the species level. The strength of this approach was demonstrated by the ability to discriminate between Thiobacillus ferrooxidans ATCC 19859 and Thiobacillus thiooxidans ATCC 8085, two very closely related species. Migration patterns for the 5S rRNA from members of the genus Thiobacillus were readily distinguishable from those of the genera Acidiphilium and Leptospirillum. In conclusion, the 5S rRNA assay represents a powerful method by which the structure of a microbial population within acidic environments can be assessed.

Characterization of a new bacteriophage which infects bacteria of the genus Acidiphilium.

A novel bacteriophage, termed phi AC1, that infects strains of the genus Acidiphilium (acidophilic, heterotrophic, aerobic, Gram-negative eubacteria) most commonly isolated from acidic mine drainage environments, has been discovered and several of its properties have been determined. This is the first report of a bacteriophage infecting such cells. The virion has a lambdoid morphology and is larger than lambda, as shown by electron microscopy and sucrose gradient centrifugation. The sedimentation coefficient of the virion is approximately 615S. The nucleic acid of phi Ac1 is dsDNA, approximately 102 kb in length. Several experimental results show that phi Ac1 is a temperate phage. The plaques are turbid, and most cells isolated from plaques produced on sensitive cells by filter-sterilized phage preparations contain the phage and are resistant to further phage infection. Southern blot analysis shows that phi Ac1 prophage DNA is integrated into the bacterial genome during the temperature growth phase.

Transformation of Acidiphilium by electroporation and conjugation.

Two techniques, electroporation and conjugation, have been used to introduce the RK2-based broad-host-range plasmids pRK415 and pLAFR3 into strains of the bacterial genus Acidiphilium. Using electroporation, cells were also transformed with a series of chimeric plasmids constructed by cloning cryptic Acidiphilium plasmids into the Escherichia coli vector pBR328. Various parameters affecting electroporation were investigated. Transformation efficiency varied widely with different recipient strains. Growth at an elevated temperature (37 degrees C) prior to electroporation increased transformation efficiency 10-fold compared with growth at 32 degrees C. For three strains tested, optimum transformation efficiency was obtained with field strengths of 10-15 kV/cm. Transformation efficiency increased linearly with increasing DNA concentration up to 10 micrograms/mL. Transformation efficiencies in these experiments ranged up to 10(4) transformants/micrograms DNA. Mobilization of pRK415 and pLAFR3 from E. coli strain S17.1 into several Acidiphilium strains was achieved following incubation for 3 h on nutrient agar medium (pH 7.0). Conjugation frequencies in the range of 10(-5)-10(-9) per recipient cell were obtained. Conjugation frequency was also dependent on recipient strain.

Relationships of structural dimensions to bench press strength in college males.

The purpose of this study was to determine the relationships between structural dimensions and bench press performance in college males. Members of required fitness classes (n = 170) were measured after 14 weeks of strength and aerobic endurance training. Anthropometric dimensions included upper arm and chest circumferences, upper and lower arm lengths, shoulder and hip widths, %fat, and height. Arm muscle cross-sectional area (CSA) was calculated from upper arm circumference corrected for triceps skinfold. Drop distance was measured from the bar to the pectoral muscles. Multiple regression analysis selected upper arm CSA, %fat, and chest circumference as the best items to predict bench press strength (R = 0.83; SEE = 11.6 kg). Cross-validation of the prediction equation on a similar sample (n = 89) produced an r = 0.74 between predicted and actual bench press (t = 0.53, p greater than 0.50). In a second cross validation sample (n = 57) who had trained more extensively with weights, the correlation between predicted and actual bench press was r = 0.57 (p less than 0.05). The prediction equations significantly (t = 6.59, p less than 0.01) underestimated bench press performance in the more extensively weight trained subjects. The results of this study suggest that bench press performance is related to structural dimensions in males and that extensive strength training may alter the relationship between size and strength.

Biodegradation kinetics of linear alkylbenzene sulfonate in sludge-amended agricultural soils.

The kinetics of ultimate biodegradation (mineralization to CO2) of linear alkylbenzene sulfonate (LAS) were studied in sludge-amended agricultural soils for a series of pure chain length LAS homologs containing 10 to 14 carbon atoms in the alkyl chain. Degradation rates were measured by following the production of 14CO2 from uniformly 14C-ring-labeled material. In general, degradation of LAS was rapid in soil over a broad concentration range (0.1 to 10 times the expected environmental concentration) and demonstrated little variation among different homologs. Half-lives for mineralization of the benzene ring ranged from 18 to 26 days and were not significantly different for any homolog over the range of alkyl chain lengths tested. Half-lives measured for LAS degradation in these studies were comparable to values reported in the literature and also to values obtained for naturally occurring materials (stearic acid, cellulose) typically present in soil environments. On the basis of the results of the present studies and those of other investigators, it is concluded that soil environments exposed to LAS in sewage sludges contain microbial communities which can actively metabolize this material. Rates of biodegradation of the benzene ring, the final step in the LAS biodegradation pathway prior to complete mineralization, are also sufficient to prevent LAS from accumulating in soil environments.

Aerobic and anaerobic biodegradation of nitrilotriacetate in subsurface soils.

Studies were conducted to characterize mineralization of nitrilotriacetate (NTA) in subsurface soils under aerobic and anaerobic conditions. Chemical (redox indicator, resazurin) and biological (dentrification) markers were used as indicators of anaerobic conditions in the test system. The indigenous microflora in subsurface soils previously exposed to septage containing NTA were able to rapidly mineralize NTA under aerobic and anaerobic conditions. The half-lives (t1/2) of mineralization for NTA in aerobic soils ranged from 87 to 160 hr. Biodegradation of NTA under anaerobic conditions where NO3- indicates that a mechanism exists for the anaerobic biodegradation of this substrate. NTA biodegradation can occur readily in the absence of molecular oxygen and the NTA-monooxygenase which are required for the aerobic mineralization of this substrate. These results provide the first evidence that the indigenous microflora in subsurface soils of septic tank tile fields can rapidly degrade NTA under anaerobic conditions.

Extrachromosomal replication of shuttle vectors in Dictyostelium discoideum.

We cloned a 12.3-kilobase (kb) endogenous plasmid, Ddp1, found in several wild-type and laboratory strains of Dictyostelium discoideum into pBR322. The cloned plasmids have been used to cotransform D. discoideum cells with B10S, a transformation vector carrying a gene fusion conferring resistance to G418. Whereas B10S DNA alone appears to integrate in a tandem array, the cloned Ddp1 plasmids replicate extrachromosomally and are stably maintained in the absence of selection with an average copy number of 50 to 100 copies per cell. The Ddp1-derived plasmids can be directly recovered by transforming Escherichia coli with bulk nuclear DNA from these cells. Preliminary deletion analysis indicates that not all regions of Ddp1 are necessary for stable replication in D. discoideum. Several recombinant vectors which replicate extrachromosomally in D. discoideum were also isolated. One contains the Act6-neor gene fusion from B10S recombined into one of the cloned derivatives of Ddp1 and can be used to directly transform D. discoideum amoebae, selecting for G418 resistance. Another recombinant is only 5.6 kb and resulted from a deletion of a 16.6-kb cloned Ddp1 hybrid plasmid. An analysis of the vector DNAs present in clones derived from single D. discoideum transformants is also described.

Intron bypass: a rapid procedure for eliminating introns from cloned genomic DNA and its application to a cellulase gene.

We have devised a DNA cloning procedure in which the introns present in a genomic DNA fragment can be eliminated easily and rapidly. The technique combines the methods of cDNA and genomic cloning in a way which assures full-length representation of the intron-free transcript. Moreover, plasmids made by this technique can be designed to contain flanking untranscribed regions which may play a role in the regulation of expression. One strand of a linearized plasmid containing the 3'-end of a gene is used to prime cDNA synthesis from an annealed mRNA template. A second plasmid containing the 5'-end of the gene is linearized, denatured, and annealed to the extended 3'-end molecules and the resulting circular, partial duplexes are used to transform bacterial cells. Two different recombinant plasmids which contain DNA encoding the cellulase, exocellobiohydrolase I, from Trichoderma reesei have been constructed using this method. They both contain the entire translated region of the gene uninterrupted by introns. One plasmid contains additional DNA at the 5'-end, including approximately 150 bp 5' to the start of transcription. The inserts of both plasmids can be excised in one piece.

Identification of an endogenous plasmid in Dictyostelium discoideum.

A plasmid has been discovered in a strain of the eukaryote, Dictyostelium discoideum, which has an unstable, non-chromosomal, cobalt resistance phenotype. The plasmid, termed Ddp1, is 13.5 kbp in size and is found in the nucleus. It has an A-T content typical of Dictyostelium DNA as judged by its restriction enzyme digestion pattern, and it is not related to either mitochondrial or ribosomal DNA. Similar or identical plasmids have been found in two original, cobalt-sensitive, isolates, NC4 and V12, but no plasmid was detected in three other isolates (WS472, WS526, WS584). The plasmid codes for non-essential functions since it is absent from the latter isolates, and it is lost from mutant strains which are capable of axenic growth.

Sediment distribution of methanogenic bacteria in lake erie and cleveland harbor.

The direct fluorescent-antibody technique was employed to determine the distribution patterns of four species of methanogens in the sediments of Lake Erie and Cleveland Harbor. Methanobacterium ruminantium was the most numerous methanogen found in regions of high-organic-silt sediments. The population of this species ranged from 10 to 10 cells/g of dry sediment. Methanobacterium strain MoH and Methanosarcina barkeri were identified in sand-silt, clay, or sand sediments. These methanogens ranged in density from 10 to 10 cells/g of dry sediment. Methanospirillum hungatii was observed only after an organic enrichment was performed on Cleveland Harbor sediments. The seasonal and selective sediment distribution of these methanogens appears to be related to the type and concentration of carbon as substrate as well as to the activities of heterotrophic and sulfate-reducing bacteria.