Christy A. Smith

Characterization of the Initial Reactions during the Cometabolic Oxidation of Methyl tert-Butyl Ether by Propane-Grown Mycobacterium vaccae JOB5

ABSTRACT The initial reactions in the cometabolic oxidation of the gasoline oxygenate, methyl tert-butyl ether (MTBE), by Mycobacterium vaccae JOB5 have been characterized. Two products, tert-butyl formate (TBF) and tert-butyl alcohol (TBA), rapidly accumulated extracellularly when propane-grown cells were incubated with MTBE. Lower rates of TBF and TBA production from MTBE were also observed with cells grown on 1- or 2-propanol, while neither product was generated from MTBE by cells grown on casein-yeast extract-dextrose broth. Kinetic studies with propane-grown cells demonstrated that TBF is the dominant (≥80%) initial product of MTBE oxidation and that TBA accumulates from further biotic and abiotic hydrolysis of TBF. Our results suggest that the biotic hydrolysis of TBF is catalyzed by a heat-stable esterase with activity toward several other tert-butyl esters. Propane-grown cells also oxidized TBA, but no further oxidation products were detected. Like the oxidation of MTBE, TBA oxidation was fully inhibited by acetylene, an inactivator of short-chain alkane monooxygenase in M. vaccae JOB5. Oxidation of both MTBE and TBA was also inhibited by propane (Ki = 3.3 to 4.4 μM). Values for Ks of 1.36 and 1.18 mM and for Vmax of 24.4 and 10.4 nmol min−1 mg of protein−1 were derived for MTBE and TBA, respectively. We conclude that the initial steps in the pathway of MTBE oxidation by M. vaccae JOB5 involve two reactions catalyzed by the same monooxygenase (MTBE and TBA oxidation) that are temporally separated by an esterase-catalyzed hydrolysis of TBF to TBA. These results that suggest the initial reactions in MTBE oxidation by M. vaccae JOB5 are the same as those that we have previously characterized in gaseous alkane-utilizing fungi.

Effects of notebook computer configuration and task on user biomechanics, productivity, and comfort

Abstract This study took a comprehensive approach to evaluating effects of using a notebook computer stand-alone or along with inexpensive peripheral input devices. The study examined effects on biomechanics, productivity, and discomfort, and considered the impact of both computer configuration and task performed. It was hypothesized that, in general, the stand-alone configuration would induce greater postural fixity and more non-neutral postures than configurations with peripheral input devices. Dependent measures included muscle activity, posture and posture variation/fixity, productivity, and subjective assessments of discomfort and preference. The data were generally consistent with the hypothesis, though some biomechanical advantages were identified for each configuration; specifics and exceptions are discussed, along with reasons for a general recommendation for the use of an external mouse, or mouse and keyboard (without number pad) when using a notebook computer for an extended period of time, as in a desktop replacement scenario. Relevance to industry Notebook computer use is rapidly increasing, in industry and schools. Yet the notebook form factor is inconsistent with a number of current design recommendations. Little research concerning physical ergonomics of notebook computer use has been conducted, so recommendations for use are currently limited and not strongly supported by objective evidence.

Cometabolism of Methyl tertiary Butyl Ether and Gaseous n-Alkanes by Pseudomonas mendocina KR-1 Grown on C5 to C8 n-Alkanes

ABSTRACT Pseudomonas mendocina KR-1 grew well on toluene, n-alkanes (C5 to C8), and 1° alcohols (C2 to C8) but not on other aromatics, gaseous n-alkanes (C1 to C4), isoalkanes (C4 to C6), 2° alcohols (C3 to C8), methyl tertiary butyl ether (MTBE), or tertiary butyl alcohol (TBA). Cells grown under carbon-limited conditions on n-alkanes in the presence of MTBE (42μ mol) oxidized up to 94% of the added MTBE to TBA. Less than 3% of the added MTBE was oxidized to TBA when cells were grown on either 1° alcohols, toluene, or dextrose in the presence of MTBE. Concentrated n-pentane-grown cells oxidized MTBE to TBA without a lag phase and without generating tertiary butyl formate (TBF) as an intermediate. Neither TBF nor TBA was consumed by n-pentane-grown cells, while formaldehyde, the expected C1 product of MTBE dealkylation, was rapidly consumed. Similar Ks values for MTBE were observed for cells grown on C5 to C8n-alkanes (12.95 ± 2.04 mM), suggesting that the same enzyme oxidizes MTBE in cells grown on each n-alkane. All growth-supporting n-alkanes (C5 to C8) inhibited MTBE oxidation by resting n-pentane-grown cells. Propane (Ki = 53 μM) and n-butane (Ki = 16 μM) also inhibited MTBE oxidation, and both gases were also consumed by cells during growth on n-pentane. Cultures grown on C5 to C8n-alkanes also exhibited up to twofold-higher levels of growth in the presence of propane or n-butane, whereas no growth stimulation was observed with methane, ethane, MTBE, TBA, or formaldehyde. The results are discussed in terms of their impacts on our understanding of MTBE biodegradation and cometabolism.

Oxidation of methyl tert-butyl ether by alkane hydroxylase in dicyclopropylketone-induced and n-octane-grown Pseudomonas putida GPo1.

ABSTRACT The alkane hydroxylase enzyme system in Pseudomonas putida GPo1 has previously been reported to be unreactive toward the gasoline oxygenate methyl tert-butyl ether (MTBE). We have reexamined this finding by using cells of strain GPo1 grown in rich medium containing dicyclopropylketone (DCPK), a potent gratuitous inducer of alkane hydroxylase activity. Cells grown with DCPK oxidized MTBE and generated stoichiometric quantities of tert-butyl alcohol (TBA). Cells grown in the presence of DCPK also oxidized tert-amyl methyl ether but did not appear to oxidize either TBA, ethyl tert-butyl ether, or tert-amyl alcohol. Evidence linking MTBE oxidation to alkane hydroxylase activity was obtained through several approaches. First, no TBA production from MTBE was observed with cells of strain GPo1 grown on rich medium without DCPK. Second, no TBA production from MTBE was observed in DCPK-treated cells of P. putida GPo12, a strain that lacks the alkane-hydroxylase-encoding OCT plasmid. Third, all n-alkanes that support the growth of strain GPo1 inhibited MTBE oxidation by DCPK-treated cells. Fourth, two non-growth-supporting n-alkanes (propane and n-butane) inhibited MTBE oxidation in a saturable, concentration-dependent process. Fifth, 1,7-octadiyne, a putative mechanism-based inactivator of alkane hydroxylase, fully inhibited TBA production from MTBE. Sixth, MTBE-oxidizing activity was also observed in n-octane-grown cells. Kinetic studies with strain GPo1 grown on n-octane or rich medium with DCPK suggest that MTBE-oxidizing activity may have previously gone undetected in n-octane-grown cells because of the unusually high Ks value (20 to 40 mM) for MTBE.

Induction of methyl tertiary butyl ether (MTBE)-oxidizing activity in Mycobacterium vaccae JOB5 by MTBE.

ABSTRACT Alkane-grown cells of Mycobacterium vaccae JOB5 cometabolically degrade the gasoline oxygenate methyl tertiary butyl ether (MTBE) through the activities of an alkane-inducible monooxygenase and other enzymes in the alkane oxidation pathway. In this study we examined the effects of MTBE on the MTBE-oxidizing activity of M. vaccae JOB5 grown on diverse nonalkane substrates. Carbon-limited cultures were grown on glycerol, lactate, several sugars, and tricarboxylic acid cycle intermediates, both in the presence and absence of MTBE. In all MTBE-containing cultures, MTBE consumption occurred and tertiary butyl alcohol (TBA) and tertiary butyl formate accumulated in the culture medium. Acetylene, a specific inactivator of alkane- and MTBE-oxidizing activities, fully inhibited MTBE consumption and product accumulation but had no other apparent effects on culture growth. The MTBE-dependent stimulation of MTBE-oxidizing activity in fructose- and glycerol-grown cells was saturable with respect to MTBE concentration (50% saturation level = 2.4 to 2.75 mM), and the onset of MTBE oxidation in glycerol-grown cells was inhibited by both rifampin and chloramphenicol. Other oxygenates (TBA and tertiary amyl methyl ether) also induced the enzyme activity required for their own degradation in glycerol-grown cells. Presence of MTBE also promoted MTBE oxidation in cells grown on organic acids, compounds that are often found in anaerobic, gasoline-contaminated environments. Experiments with acid-grown cells suggested induction of MTBE-oxidizing activity by MTBE is subject to catabolite repression. The results of this study are discussed in terms of their potential implications towards our understanding of the role of cometabolism in MTBE and TBA biodegradation in gasoline-contaminated environments.

An investigation of ergonomic interventions in dental hygiene work.

Alternative methods for viewing teeth while performing simulated dental procedures were investigated. The methods allowed participants to assume postures requiring less neck flexion than the standard direct view. One alternative used a video camera and monitor to view the mouth, the other incorporated 90 degrees prism glasses. The study was conducted in two parts: (1) novice participants performing a targeting task; (2) dental hygienists performing a scaling task on a mouth model. Posture and subjective perceptions were assessed in Parts 1 and 2. Muscle activity and performance were also assessed in Part 1. The alternative methods significantly reduced muscle activity, neck flexion, and discomfort, compared to the direct view. Preferences were a function of criteria (general, comfort, productivity, or accuracy). Previously, recommendations for reducing ergonomic risk factor exposure of dental professionals emphasized reducing time spent performing dental procedures. This study shows ergonomic interventions offer alternative means of risk exposure reduction.

Ergonomic interventions for the furniture manufacturing industry. Part II -- Handtools

Abstract The objectives of this intervention research project were to develop and evaluate engineering controls for the reduction of the upper extremity injury risk in workers in the furniture manufacturing industry. The analysis of OSHA Form 200 logs and surveys of furniture workers revealed that upholsterers, workers who use random orbital sanders and workers who use spray guns are at higher levels of risk of illness than the rest of the working population. An on-site ergonomic analysis of these three jobs was performed and the following risk factors were identified for each of these three work groups: upholsterers—repetitive, high-force pinch grips; sanders—long-duration static grip forces; and sprayers—awkward postures (ulnar wrist deviations and wrist flexion). Engineering interventions in the form of new or modified handtools were then evaluated in the laboratory to assess their effectiveness in reducing exposure to these risk factors. For sanding, an interface was created that secured the hand to the sander with the intention of reducing the need for static grip forces during sanding. A new handtool was created for upholsterers that replaced the repetitive pinch grips with a power grip. Finally, a commercially available spray gun with ergonomic features was evaluated. Each of these modified tools/methods was compared with the standard methods typically used in industry. The results show that most of the intended beneficial effects were realized. The random orbital sander interface reduced extensor muscle activities by an average of 30%. The upholstery handtool reduced the intrinsic hand muscle activities by an average of 51%. The effects of the adapted spray gun were most prominent when working on horizontal surfaces and showed an average reduction of 40° of wrist flexion and 14° of ulnar deviation as compared to the standard pistol grip spray gun in this activity. Relevance to industry The ergonomic intervention research described in this report documents a reduction in exposure to risk factors for upper extremity cumulative trauma disorders for three work activities in the furniture manufacturing industry.

Technical Note The use of mirrors during an assembly task: a study of ergonomics and productivity

Industrial assembly tasks often require awkward, sustained neck and/or shoulder postures that can lead to increased musculoskeletal discomfort and reduced productivity. The aim of this study was to investigate the effects of mirror and periscope visual aids as ergonomic interventions designed to eliminate awkward postures of the cervicobrachial region during assembly tasks. Participants simulated a simple assembly task by using a cordless screwdriver to drive screws into a pre-tapped aluminium block. Trials of 15 min were run for each of four distinct assembly workstation configurations: industry standard (in-line screwdriver, work at elbow height, no visual aid); pistol grip (pistol grip screwdriver, work at shoulder height, no visual aid); mirror (in-line screwdriver, work at elbow height, single mirror visual aid); and periscope (in-line screwdriver, work at elbow height, two-mirror visual aid system). Muscular activity, discomfort, body posture, productivity and operator subjective assessment were recorded to determine the effects of the visual aid interventions. The results show that when comparing the interventions to the industry standard condition, there was a 45% reduction in average cervical erector spinae activity, a 90% reduction in average neck flexion angle and a 72% reduction in neck discomfort with the interventions. When comparing these interventions to the pistol grip condition there was an 80% reduction in activity of the dominant side deltoid, a 92% reduction in shoulder flexion angle and an 81% decrease in shoulder discomfort with the interventions. Productivity was greatest in the industry standard configuration followed by the pistol grip (9% lower), the periscope (13% lower) and the mirror (23% lower) configurations. A follow-up study that compared the productivity of the periscope configuration with that of the industry standard configuration showed that within a 4-h work period this productivity differential decreased by over 33%.

Ergonomic interventions for the furniture manufacturing industry. Part I -- Lift assist devices

Abstract The objectives of this intervention research project were to develop and evaluate engineering controls for the reduction of low back injury risk in workers in the furniture manufacturing industry. An analysis of injury/illness records and survey data identified upholsterers and workers in the machine room as two occupations within the industry at elevated risk for low back injury. A detailed ergonomic evaluation of the activities performed by these workers was then performed and the high risk subtasks were identified. The analysis for upholsterers revealed: (1) high forces during the loading and unloading of the furniture to and from the upholstery bucks, (2) static awkward postures ( extreme flexion>50° , lateral bending>20° , twisting>20°) during the upholstering of the furniture, and (3) repetitive bending and twisting throughout the operation. For machine room workers, this ergonomic evaluation revealed repetitive bending and twisting (up to 5 lifts/min and sagittal flexion>80°, lateral bending>15°, twisting>45°) when getting wooden components from or moving them to the shop carts that are used to transport these materials. Engineering interventions were then developed and evaluated in the laboratory to document the reduction of exposure to these stressors. The height-adjustable upholstery buck system eliminated the lifting and lowering requirements and affected trunk kinematics during the upholstery operation by reducing peak sagittal angles by up to 79% (average: 52%; range: 27–79%), peak sagittal accelerations by up to 42% (average: 71%; range: 0–74%) and peak lateral position by up to 31% (average: 20%; range: 12–31%), and showed no impact on time to complete the task. The machine room lift reduced peak sagittal angle by up to 90% (average: 76%; range: 64–90%), peak sagittal accelerations by up to 86% (average: 72%; range: 59–86%) and had a positive impact on the time to complete the task (average reduction: 19%). Relevance to industry The ergonomic intervention research documented in this report shows the impact of engineering controls for the furniture manufacturing industry on the risk factors for work-related low back injuries.

Oxidation of gasoline oxygenates by closely related non-haem-iron alkane hydroxylases in Pseudomonas mendocina KR1 and other n-octane-utilizing Pseudomonas strains.

Pseudomonas mendocina KR1 oxidizes the gasoline oxygenate methyl tertiary butyl ether (MTBE) to tertiary butyl alcohol (TBA) during growth on C5 -C8 n-alkanes. We have further explored oxidation of ether oxygenates by this strain to help identify the enzyme that catalyses these reactions. High levels of MTBE-oxidizing activity occurred in resting cells grown on C5 -C8 n-alkanes. Lower activities occurred in cells grown on longer-chain n-alkanes (C9 -C11 ) and 1°-alcohols (C5 -C10 ). N-octane-grown cells also oxidized tertiary amyl methyl ether (TAME) to tertiary amyl alcohol (TAA), but did not oxidize ethyl tertiary butyl ether (ETBE), TBA or TAA. A 39 kDa polypeptide in whole cell extracts of n-octane-grown cells strongly cross-reacted with an anti-AlkB polyclonal antiserum in an SDS-PAGE/immunoblot. This polypeptide was absent or less abundant in cells grown on dextrose, dextrose plus dicyclopropylketone or 1-octanol. N-octane-grown cells of Pseudomonas aeruginosa strains KSLA-473 and ATCC 17423 oxidized MTBE and TAME but not ETBE. N-hexadecane-grown cells of these strains and strain PAO1 did not oxidize any of the oxygenates tested. Our results indicate ether oxygenate-degrading activity in alkane-utilizing pseudomonads is consistently observed with close homologues of the GPo1 non-haem-iron alkane hydroxylases but is otherwise not a consistent catalytic feature of these diverse enzymes.