Enhanced mating-type switching and sexual hybridization in heterothallic yeast Yarrowia lipolytica.

Yarrowia lipolytica is a non-conventional, heterothallic, oleaginous yeast with wide range of industrial applications. Increasing ploidy can improve advantageous traits for industrial applications including genetic stability, stress resistance, and productivity, but the construction of knockout mutant strains from polyploid cells requires significant effort due to the increased copy numbers of target genes. The goal of this study was to evaluate the effectiveness of a mating-type switching strategy by single-step transformation without a genetic manipulation vestige, and to optimize the conventional method for increasing ploidy (mating) in Y. lipolytica. In this study, mating-type genes in haploid Y. lipolytica cells were scarlessly converted into the opposite type genes by site-specific homologous recombination, and the resulting MATB-type cells were mated at low temperature (22°C) with addition of sodium citrate with each MATA-type haploid cell to yield a MATA/MATB-type diploid strain with genetic information from both parental strains. The results of this study can be used to increase ploidy and for whole genome engineering of a yeast strain with unparalleled versatility for industrial application.

Information on functioning found in the medical records of patients with stroke.

[Purpose] To explore data on functioning in the medical records of patients with stroke by linking them to the ICF. [Subjects and Methods] The admission and discharge summaries of patients' electronic medical records (EMRs) were investigated. Information on functioning included in the summaries were mapped into the ICF. Each of the linked categories of ICF was analyzed using frequency and percentage analysis. [Results] Thirty stroke patients' records were evaluated. A total of 1,832 items on functioning in the EMRs were found to be linked to eighty-five categories of the ICF. The majority of those categories (52.9%) belonged to the body function domain, whereas only 8.2% were environment factors. Categories in the domain of activity and participation, and body structure were found to be 22.4% and 16.5% respectively. In each domain, the most frequently found categories were muscle power function (b730), structure of brain (s110), walking (d450), and products or substances for personal consumption (e110). [Conclusion] It was found that the admission and discharge summary in the current medical records of patients with stroke contained much noticeable information on functioning and the data on functioning may be linked to the ICF. Further study is needed to adapt ICF in Korean clinical settings.

Fitness costs of rifampicin resistance in Mycobacterium tuberculosis are amplified under conditions of nutrient starvation and compensated by mutation in the ß' subunit of RNA polymerase.

Rifampicin resistance, a defining attribute of multidrug-resistant tuberculosis, is conferred by mutations in the ß subunit of RNA polymerase. Sequencing of rifampicin-resistant (RIF-R) clinical isolates of Mycobacterium tuberculosis revealed, in addition to RIF-R mutations, enrichment of potential compensatory mutations around the double-psi ß-barrel domain of the ß' subunit comprising the catalytic site and the exit tunnel for newly synthesized RNA. Sequential introduction of the resistance allele followed by the compensatory allele in isogenic Mycobacterium smegmatis showed that these mutations respectively caused and compensated a starvation enhanced growth defect by altering RNA polymerase activity. While specific combinations of resistance and compensatory alleles converged in divergent lineages, other combinations recurred among related isolates suggesting transmission of compensated RIF-R strains. These findings suggest nutrient poor growth conditions impose larger selective pressure on RIF-R organisms that results in the selection of compensatory mutations in a domain involved in catalysis and starvation control of RNA polymerase transcription.

Surface EMG during the Push-up plus Exercise on a Stable Support or Swiss Ball: Scapular Stabilizer Muscle Exercise.

[Purpose] Scapular stabilizer strengthening exercise is crucial for shoulder rehabilitation. The purpose of this study was to compare two types of push-up plus exercises, on a stable and unstable bases of support, using surface electromyography (EMG), to suggest an effective shoulder rehabilitation program. [Subjects and Methods] Ten healthy men volunteered for this study. All volunteers performed two sets of push-up plus exercise (standard push up and knee push up) on stable and unstable bases of support. The muscle activities of five important scapular stabilizer muscles (upper trapezius, middle trapezius, lower trapezius, serratus anterior, latissimus dorsi) were recorded during the exercise. [Results] The upper trapezius showed greater mean electric activation amplitude in the scapular retraction posture than in the scapular protraction posture, and the serratus anterior showed greater mean electric activation amplitude in the scapular protraction posture than in the scapular retraction posture. The root-mean-square normalized EMG values of the muscles were greater during the exercise performed on the unstable support than those on the stable support. [Conclusion] The standard push-up plus exercise on an unstable base of support helps to increase muscle activity, especially those of the upper/middle trapezius and serratus anterior.

Amperometric Sensing of Carbon Monoxide: Improved Sensitivity and Selectivity via Nanostructure-Controlled Electrodeposition of Gold.

A series of gold (Au) nanostructures, having different morphologies, were fabricated for amperometric selective detection of carbon monoxide (CO), a biologically important signaling molecule. Au layers were electrodeposited from a precursor solution of 7 mM HAuCl4 with a constant deposition charge (0.04 C) at various deposition potentials. The obtained Au nanostructures became rougher and spikier as the deposition potential lowered from 0.45 V to 0.05 V (vs. Ag/AgCl). As prepared Au layers showed different hydrophobicity: The sharper morphology, the greater hydrophobicity. The Au deposit formed at 0.05 V had the sharpest shape and the greatest surface hydrophobicity. The sensitivity of an Au deposit for amperometric CO sensing was enhanced as the Au surface exhibits higher hydrophobicity. In fact, CO selectivity over common electroactive biological interferents (L-ascorbic acid, 4-acetamidophenol, 4-aminobutyric acid and nitrite) was improved eminently once the Au deposit became more hydrophobic. The most hydrophobic Au was also confirmed to sense CO exclusively without responding to nitric oxide, another similar gas signaling molecule, in contrast to a hydrophobic platinum (Pt) counterpart. This study presents a feasible strategy to enhance the sensitivity and selectivity for amperometric CO sensing via the fine control of Au electrode nanostructures.

Production of Bio-Based Isoprene by the Mevalonate Pathway Cassette in Ralstonia eutropha.

Isoprene has the potential to replace some petroleum-based chemicals and can be produced through biological systems using renewable carbon sources. Ralstonia eutropha can produce value-added compounds, including intracellular polyhydroxyalkanoate (PHA) through fatty acid and lipid metabolism. In the present study, we engineered strains of R. eutropha H16 and examined the strains for isoprene production. We optimized codons of all the genes involved in isoprene synthesis by the mevalonate pathway and manipulated the promoter regions using pLac and pJ5 elements. Our results showed that isoprene productivity was higher using the J5 promoter (1.9 ± 0.24 µg/l) than when using the lac promoter (1.5 ± 0.2 µg/l). Additionally, the use of three J5 promoters was more efficient (3.8 ± 0.18 µg/l) for isoprene production than a one-promoter system, and could be scaled up to a 5-L batch-cultivation from a T-flask culture. Although the isoprene yield obtained in our study was insufficient to meet industrial demands, our study, for the first time, shows that R. eutropha can be modified for efficient isoprene production and lays the foundation for further optimization of the fermentation process.