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mature carrot (Daueus carota) roots and purification and characterization of a novel isoform. -Physiol. Plant. 96: 130-138,The profile of cMtinases (EC 3.2.1,14) in mature carrot {Daucus carota L. cv. Eagle) roots was studied. Multiple chitinase bands (8-10) were observed in native and sodium dodecylsulfate-denaturing polyacrylamide gels. The molecular masses of these chitinases were estimated to be from 20 000 to 40 000. One major chitinase was purified and found to be an acidic protein with pi at 4,3 and a molecular mass of 39 500. The optimum pH for enzymatic activity was around 5 and the optimum temperature was 25°C, The enzyme was stable at pHs below 8 and temperatures below 60°C. The protein did not have a chitin-binding domain, but showed some similarity to the amino acid composition of tobacco class I chitinase. The N-terminal amino acid sequence did not resemble any ofthe described classes of chitinases. This chitinase did not possess lysozyme activity and showed antifungal activity when tested against Trichoderma sp.

The profile of chaitinases (EC 3.2.1.14) in mature carrot (Daucus carota L. cv. Eagle) roots was studied. Multiple chitinase bands (8–10) were observed in native and sodium dodecylsulfate‐denaturing polyacrylamide gels. The molecular masses of these chitinases were estimated to be from 20 000 to 40 000. One major chitinase was purified and found to be an acidic protein with pI at 4.3 and a molecular mass of 39 500. The optimum pH for enzymatic activity was around 5 and the optimum temperature was 25°C. The enzyme was stable at pHs below 8 and temperatures below 60°C. The protein did not have a chitin‐binding domain, but showed some similarity to the amino acid composition of tobacco class I chitinase. The N‐terminal amino acid sequence did not resemble any of the described classes of chitimases. This chitinase did not possess lysozyme activity and showed antifungal activity when tested against Trichoderma sp.

Xylose reductase catalyzes the reduction of xylose to xylitol and is known to play a pivotal role in pentose metabolism in yeasts. We previously showed that a cystein residue may be involved in binding of the coenzyme NADPH to the Pichia stipitis xylose reductase through chemical modification studies. The question arose as to which of the three cysteine residues in this enzyme may be involved in coenzyme binding. We cloned the XYL1 gene encoding xylose reductase from P. stipitis into the phagemid pEMBL18(+) suitable for site-directed mutagenesis. Each of the three cysteine residues (Cys19, Cys27 and Cys130) was individually mutated to serine. All three Cys-->Ser variants remained functional, but with reduced catalytic activity. Sensitivity of the P. stipitis xylose reductase to thiol-specific reagents was attributed to both Cys27 and Cys130 residues as substitution of either residue with Ser resulted in a significant but incomplete loss of sensitivity to PCMBS. The apparent Km values of the Cys variants for NADPH, NADH and xylose did not differ from those of the wild-type enzyme isolated from yeast by more than 4-fold. Our results suggest that none of the Cys residues are directly involved in NADPH binding, although Cys130 may reside in or near the coenzyme binding region and might play a role in coenzyme specificity.

BackgroundPhysician dissatisfaction with more time spent on related paperwork but less time available for direct interaction with patients is increasing internationally. Increased physician workload resulting from paperwork might negatively affect their interaction with patients and increase the risk for burnout. This study aimed to investigate the level of physician workload tethered to paperwork during outpatient encounters and explore its latent workload subgroups among Chinese physicians.MethodsA cross-sectional survey was conducted via online questionnaire primarily in 24 hospitals in 6 provinces in Eastern, Central, and Western China from November 2020 to February 2021. The Chinese physician mental workload scale developed by our research team was used for assessment of physician workload tethered to paperwork. Physicians were categorized into different subgroups of workload via latent profile analysis. Multinomial logistic regression was subsequently performed to examine how demographic variables differ among physicians belonging to different subgroups.ResultsA total of 1,934 valid questionnaires were received. Chinese physicians reported medium levels of workload while performing non-physician-patient communication work tasks characterized by paperwork during outpatient encounters. Four latent workload subgroups were identified: “low workload group” (8.8%), “medium workload group” (34.0%), “high workload group” (42.1%) and “very high workload group” (15.1%). Compared with the other latent workload subgroups, physicians belonging to the “very high workload group” were more likely to be younger, married, those who had worse health status, lower educational levels and lower average monthly incomes, those who worked more years in the current institution, more hours per week and longer outpatient hours per week, and those who worked in public general hospitals, tertiary B hospitals and Obstetrics and Gynecology, and saw more than 50 outpatients per day, with more time spent on per patient.ConclusionsThere exit four latent workload subgroups among Chinese physicians tethered to paperwork during outpatient encounters along with great individual variations among these subgroups. The characteristics of the latent “very high workload group” can help permit more targeted guidance for developing interventions with optimized human resource allocation to, in turn, increase the time available for direct interaction with patients, thereby resulting in improved quality of physician-patient interactions and decreased risk for physician burnout.