Improvement of the recombinant phytase expression by intermittent feeding of glucose during the induction phase of methylotrophic yeast Pichia pastoris.

PURPOSE: For growth of methylotrophic yeast, glycerol is usually used as a carbon source. Glucose is used in some cases, but not widely consumed due to strong repressive effect on AOX1 promoter. However, glucose is still considered as a carbon source of choice since it has low production cost and guarantees growth rate comparable to glycerol. RESULTS: In flask cultivation of the recombinant yeast, Pichia pastoris GS115(pPIC9K-appA38M), while methanol induction point(OD600) and methanol concentration significantly affected the phytase expression, glucose addition in induction phase could enhance phytase expression. The optimal flask cultivation conditions illustrated by Response Surface Methodology were 10.37 OD600 induction point, 2.02 h before methanol feeding, 1.16% methanol concentration and 40.36µL glucose feeding amount(for 20 mL culture volume) in which the expressed phytase activity was 613.4 ± 10.2U/mL, the highest activity in flask cultivation. In bioreactor fermentation, the intermittent glucose feeding showed several advantageous results such as 68 h longer activity increment, 149.2% higher cell density and 200.1% higher activity compared to the sole methanol feeding method. These results implied that remaining glucose at induction point might exhibit a positive effect on the phytase expression. CONCLUSION: Glucose intermittent feeding could be exploited for economic phytase production and the other recombinant protein expression by P. pastoris GS115.

Betterment of a displacement retrieval method based on data fitting in a near-half-fringe zone.

Raising the speed and accuracy of displacement retrieval by overcoming the weakness of time-consuming parameter estimation and a complex displacement retrieval algorithm is of key importance in improving the real-time quality of optical feedback self-mixing-based displacement measurement sensor. This paper proposes a high-speed and accurate simultaneous estimation method of displacement, optical feedback factor, and linewidth enhancement factor based on data fitting near a half-fringe cooperating with the fringe-counting method. The displacement measurement accuracy of the proposed method is about 16 nm. The high estimation speed ensured by the principle of the reported method constitutes the main character of the method; besides, the method can be applied to any motion, as well as a harmonic vibration.

Characteristics and Effectiveness of Mobile- and Web-Based Tele-Emergency Consultation System between Rural and Urban Hospitals in South Korea: A National-Wide Observation Study.

(1) Background: The government of South Korea has established a nationwide web- and mobile-based emergency teleconsultation network by designating urban and rural hospitals. The purpose of this study is to analyze the characteristics and effectiveness of the tele-emergency system in South Korea. (2) Methods: Tele-emergency consultation cases from May 2015 to December 2018 were analyzed in the present study. The definition of a tele-emergency in the present study is an emergency consultation between doctors in rural and urban hospitals via a web- and mobile-based remote emergency consultation system (RECS). Consultations through an RECS are grouped into three categories: medical procedure or treatment guidance, image interpretation, and transportation requests. The present study analyzed the characteristics of the tele-emergency system and the reduction in unnecessary transportation (RUT). (3) Results: A total of 2604 cases were analyzed in the present study from 2985 tele-emergency consultation cases. A total of 381 cases were excluded for missing data. Consultations for image interpretation were the most common in trauma cases (71.3%), while transfer requests were the most common in non-trauma cases (50.3%). Trauma patients were more frequently admitted to rural hospitals or discharged and followed up with at rural hospitals (20.3% vs. 40.5%) after consultations. In terms of disease severity, non-severe cases were statistically higher in trauma cases (80.6% vs. 59.4%; p < 0.001). The RUT was statistically highly associated with trauma cases (60.8% vs. 42.8%; p < 0.001). In an analysis that categorized cases by region, a statistically higher proportion of transportation was used in island regions (69.9% vs. 49.5%; p < 0.003). More RUT was associated with non-island regions (30.1% vs. 50.5%; p = 0.001). (4) Conclusions: The tele-emergency system had a great role in reducing unnecessary patient transportation in non-severe trauma cases and non-island rural area emergency cases. Further research is needed for a cost/benefit analysis and clinical outcomes.

Multidisciplinary integrated optimal design process for optomechanical structures.

This paper presents an integrated design process for optomechanical structures based on multidisciplinary optimization. The proposed integrated optimal design process comprises a finite element analysis by ANSYS Workbench, the MATLAB optomechanical transfer program, an optical analysis by ZEMAX, and the multidisciplinary optimization solver by Isight. In ANSYS Workbench, the deformation of optical surfaces, structures, and responses according to the design requirements is calculated in one project. Then, Zernike polynomial coefficients are calculated from surface deformation data of optical surfaces through a MATLAB optomechanical transfer program. In ZEMAX, the Zernike polynomial coefficients are imported into optical surface models of an optical system; then, optical performance parameters, such as the wavefront error, optical aberration, MTF, and OPD, are calculated. In the Isight environment, automatic iterative computations are performed between these three programs and, as a result, the design dimensions of optomechanical structures are determined, satisfying the design requirements and improving the performance of an optical system. By using this integrated optimal design process, the optimal design and analysis for a complete optomechanical structure, as well as individual structure parts, can be performed successfully. In this paper, the optimal design problem for three parts of a Cassegrain telescope, which consists of a primary mirror with an outer diameter of 156 mm and a secondary mirror with an outer diameter of 46 mm, was taken as an example. By using optimal parts, the image wavefront error of the Cassegrain telescope was decreased from 29.9 to 16.1 nm.

Lateral position does not cause an interhemicerebral difference of cerebral hemodynamic in healthy adult volunteers.

Cerebral perfusion is maintained at a consistent value irrespective of changes in systemic blood pressure or disease-induced changes in general physical condition. This regulatory mechanism is effective despite postural changes, working even during changes in posture, such as those from sitting to standing or from the head-down to the head-up position. However, no study has addressed changes in perfusion separately in the left and right cerebral hemispheres, and there has been no specific investigation of the effect of the lateral decubitus position on perfusion in each hemisphere. Surgery, particularly respiratory surgery, is often performed with the patient in the lateral decubitus position, and since intraoperative anesthesia may also have an effect, it is important to ascertain the effect of the lateral decubitus position on perfusion in the left and right cerebral hemispheres in the absence of anesthesia. The effects of the lateral decubitus position on heart rate, blood pressure, and hemodynamic in the left and right cerebral hemispheres assessed by regional saturation of oxygen measured by near-infrared spectroscopy were investigated in healthy adult volunteers. Although the lateral decubitus position causes systemic circulatory changes, it may not cause any difference in hemodynamic between the left and right cerebral hemispheres.

Effects of cloth face masks on physical and cognitive performance during maximal exercise testing.

Wearing a cloth face mask has been shown to impair exercise performance; it is essential to understand the impact wearing a cloth face mask may have on cognitive performance. Participants completed two maximal cardiopulmonary exercise tests on a cycle ergometer (with and without a cloth face mask) with a concurrent cognitive task. Blood pressure, heart rate, oxygen saturation, perceived exertion, shortness of breath, accuracy, and reaction time were measured at rest, during each exercise stage, and following a 4-minute recovery period. The final sample included 35 adults (age = 26.1 ± 5.8 years; 12 female/23 male). Wearing a cloth face mask was associated with significant decreases in exercise duration (-2:00 ± 3:40 min, P = 0.003), peak measures of maximal oxygen uptake (-818.9 ± 473.3 mL/min, -19.0 ± 48 mL·min-1·kg-1, P < 0.001), respiratory exchange ratio (-0.04 ± 0.08, P = 0.005), minute ventilation (-36.9 ± 18 L/min), oxygen pulse (-3.9 ± 2.3, P < 0.001), heart rate (-7.9 ± 12.6 bpm, P < 0.001), oxygen saturation (-1.5 ± 2.8%, P = 0.004), and blood lactate (-1.7 ± 2.5 mmol/L, P < 0.001). While wearing a cloth face mask significantly impaired exercise performance during maximal exercise testing, cognitive performance was unaffected in this selected group of young, active adults.

Normalization-free displacement reconstruction method based on fringe scaling.

The knotty problems in the displacement reconstruction based on the self-mixing (SM) technique are the estimation of the self-mixing interferometry parameters and the normalization of SM signals (SMSs) since they are all very time-consuming and based on complex algorithms. This has an unfavorable effect on the real-time and low-cost natures of the SM displacement sensor. In the paper, we have presented a simple method of displacement retrieval with a high resolution, which does not require the parameter estimation and normalization. The proposed method is based on the scaling of individual fringes in SMSs and speckle noise proof. The simplicity of the method will make a great contribution to lowering the cost of the SM displacement sensor and improving the reliability and resolution of the sensor.

Paraconexibacter antarcticus sp. nov., a novel actinobacterium isolated from Antarctic tundra soil.

A Gram-negative, non-motile, aerobic bacterium, named 02-257T, was isolated from Antarctic soil. The cells are surrounded by relatively thin capsules and were catalase-positive and oxidase-negative cocci. Growth of strain 02-257T was observed at 4-35 °C (optimum, 28-30 °C), pH 6.0-8.0 (optimum, pH 6.0) and with 0-1.5% NaCl (optimum, 0 %). Strain 02-257 showed the highest 16S rRNA gene sequence similarity to Paraconexibacter algicola Seoho-28T (95.06 %). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain 02-257T is a member of a novel species belonging to the clade formed by members of the genus Paraconexibacter in the family Paraconexibacteraceae. The DNA G+C content was 72.9 mol%. Strain 02-257T contained C16 : 0-iso (23.0 %), C18  :  1 ω9c (13.8 %), C16 : 0 (12.5 %) and C17 : 1 ω9c-iso (10.8 %) as major cellular fatty acids and menaquinone MK-7(H4) was detected as the only isoprenoid quinone. Diphosphatidylglycerol, phosphatidylinositol, phosphatidylinositole mannoside, phosphatidylinositole dimannoside, unidentified phosphoglycolipid, unidentified aminophospholipid, two unidentified phospholipids, three unidentified aminolipids and six unidentified lipids were the major polar lipids. meso-Diaminopimelic acids were the diagnostic diamino acids in the cell-wall peptidoglycan. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain 02-257T is considered to represent a novel species of the genus Paraconexibacter, for which the name Paraconexibacter antarcticus sp. nov. is proposed. The type strain is 02-257T (=CCTCC AB 2021030T=KCTC 49619T).

The wavevector star channel and symmetry group.

The concepts of `wavevector star channel' and `wavevector star channel group' are newly defined, which allow the effective study of phase transitions considering directly the translational symmetry breaking in crystals. A method is suggested by which the wavevector star channels can be found using the image of the representation of the translational group. According to this method, the wavevector star channels are found for the 80 Lifschitz stars in the reciprocal lattice. The wavevector star channel group is defined as the set of symmetry elements of the parent phase which leave the star channel invariant, and the wavevector star channel groups with one, two, three and four arms are calculated. It is shown that the complicated symmetry changes in the pyroelectric crystal Pb1-xCaxTiO3 (PCT) can be described using the new five-component reducible order parameter transformed according to the representation of the wavevector star channel group, rather than the nine-component one based on the theory of the full irreducible representation of the space group.

Verification of phase measurement error sources in phase-shifting interferometry with four step phase-shifting algorithms.

The phase-shifting technique is one of the most important techniques in interferometry. The measurement accuracy of this technique is affected by several kinds of systematic and random errors like miscalibration, nonlinear response and tilt shift of the phase shifter, nonlinear recording, speckles, fringe fluctuation, etc. In this paper, a method for verification of all of these phase measurement error sources is proposed by introducing a grayscale image. This image is composed from the lattice-site representation determined by a four step phase-shifting algorithm, which represents the distribution of phase-shift angle values. The shape of the grayscale image is definitely more expressive and demonstrative for the kinds of measurement error sources than the black dot distribution in lattice-site representation. So, with this technique, we can recognize the existence of various measurement error sources and roughly estimate their magnitudes before proceeding with experiments. Our method is to be rather qualitative than quantitative. The availability of this method is discussed in a phase-shifting Fizeau interferometer and a fringe projection profilometry. This method is useful in all kinds of interferometric measurements based on the phase-shifting technique.

Joint estimation of self-mixing interferometry parameters and displacement reconstruction based on local normalization.

It is not easy to estimate self-mixing interferometry parameters, namely, the optical feedback factor and the linewidth enhancement factor from the self-mixing signals (SMSs) affected by noise such as speckle. These SMSs call for normalization, which is not only difficult, but also apt to distort the intrinsic information of the signals, thereby resulting in incorrect estimation of the parameters and the displacement reconstruction. In this paper, we present what we believe is a novel normalization method we call "local normalization," which enables more exact and simpler estimation and displacement retrieval compared to previous methods, for it is based on an analytic relation instead of approximation. The method is very noise-proof, and especially speckle-noise-proof as well. The method proposed can be applied to moderate and strong feedback regimes. The simplicity and accuracy of the method will provide a fine tool for a low-cost self-mixing displacement sensor with a high resolution of about 40 nm.

Preventive effect of subacidic 1% NaF-HF gel on dental caries in 6- to 7-year-old schoolchildren: a randomized controlled trial.

Objective: This study aimed to examine and compare the anti-caries effects of slightly acidic 1% NaF-HF gel when applied once or twice (7-day or 6-month interval) to mandibular first molars in 6- to 7-year-old children. Method and materials: This was a 1-year, multi-arm, placebo-controlled, double-blind, parallel-group randomized trial. In total, 1,077 schoolchildren from eight primary schools were allocated to one of the four study groups (group 1, single application; group 2, two applications with a 7-day interval; group 3, two applications with a 6-month interval; group 4, placebo control). The occlusal surfaces of permanent mandibular first molars were examined by three calibrated examiners who were blind to the group allocation at baseline and the end of the study. Caries reduction and prevented fraction was assessed. Results: A total of 986 children completed the study. There was a statistically significant difference in the mean D1MFT increment across the groups (P < .001). The mean D1MFT increment was 0.37 in group 1 (single application), 0.18 in group 2 (two applications with a 7-day interval), 0.21 in group 3 (two applications with a 6-month interval), and 0.56 in the control group. The mean D2MFT increments (0.22 in group 1, 0.06 in group 2, 0.18 in group 3, and 0.50 in the control group) also showed a statistically significant difference across the groups (P < .001). Conclusion: Subacidic 1% NaF-HF gel exerts a high preventive effect on caries (more than 40%) when applied twice at a 7-day interval in 6- to 7-year-old schoolchildren who do not use fluoride toothpaste in areas where drinking water is not fluoridated.

Reduction of peroxynitrite by some manganoporphyrins of AEOL series: DFT approach with dispersion correction and NBO analysis.

The AEOL series of manganoporphyrins (MnP; AEOL compounds were named by US Aeolus pharmaceuticals) designed as superoxide dismutase mimic are well-known for their powerful catalytic activity to neutralize reactive oxygen and nitrogen species. Reductive oxygen atom cleavage from peroxynitrite (ONOO-) to form NO2 in aqueous solution by some AEOL compounds (AEOL-10113, AEOL-10150, AEOL-11114 and AEOL-11203) was studied by DFT/M06-2X computations with D3 dispersion correction and gCP (geometrical counterpoise correction) for basis set superposition error. DFT computation showed that AEOL-10150 can form the most stable association complex {MnP…OONO} among four AEOL models. AEOL-10150 complex with ONOO- has the lowest deformation energy. In AEOL compounds and their association complexes with ONOO-, Mn atom prefered the high spin state (S = 2) to the intermediate spin state (S = 1). Natural bond orbital analysis showed that electron transfer from the most negative oxygen atom in ONOO- to Mn atom in MnP has the biggest interaction energy among all kinds of donor-acceptor interactions between ONOO- and MnP.

Fast estimation method of feedback factor based on the derivative of the self-mixing signal.

The self-mixing (SM) technique has been a promising optical measurement technique for a few decades. The estimation of the optical feedback factor C is the key to SM displacement measurement with a better resolution than half a wavelength. But this estimation is extremely complex and tricky, and thus leads to its time-consuming nature, which is one of the important causes that increases the cost and labor for the development of a self-mixing sensor. There have been many methods reported for the C estimation, but they are very time consuming, very sensitive to noise, or can only work in a certain feedback regime. This paper presents a novel approach to the C estimation, which is straightforward and can work for all feedback regimes including weak, moderate, and strong. The estimation precision of the proposed method is not only comparable to that of the data fitting technique but also much simpler.

Evaluation method for the optical feedback factor and linewidth enhancement factor using phase discontinuities in self-mixing interferometry signals.

Self-mixing interferometry (SMI) is a reliable method that has been applied to measuring displacements, absolute distances, and velocities of remote targets. Evaluating the optical feedback factor C and the linewidth enhancement factor α is a vital step in calculating laser diode parameters and in processing SMI signals using phase unwrapping. This paper proposes an evaluation method for the optical feedback factor and the linewidth enhancement factor of arbitrary waveforms by investigating the slopes of phase discontinuity distribution in the optical feedback regime of 1

The KLF6 Super Enhancer Modulates Cell Proliferation via MiR-1301 in Human Hepatoma Cells.

BACKGROUND: Recent studies have attempted to elucidate the function of super enhancers by means of microRNAs. Although the functional outcomes of miR-1301 have become clearer, the pathways that regulate the expressions of miR-1301 remain unclear. OBJECTIVE: The objective of this paper was to consider the pathway regulating expression of miR- 1301 and miR-1301 signaling pathways with the inhibition of cell proliferation. METHODS: In this study, we prepared the cell clones that the KLF6 super enhancer was deleted by means of the CRISPR/Cas9 system-mediated genetic engineering. Changes in miR-1301 expression after the deletion of the KLF6 super enhancer were evaluated by RT-PCR analysis, and the signal pathway of miR-1301 with inhibition of the cell proliferation was examined using RNA interference technology. RESULTS: The results showed that miR-1301 expression was significantly increased after the deletion of the KLF6 super enhancer. Over-expression of miR-1301 induced by deletion of the KLF6 super enhancer also regulated the expression of p21 and p53 in human hepatoma cells. functional modeling of findings using siRNA specific to miR-1301 showed that expression level changes had direct biological effects on cellular proliferation in Human hepatoma cells. Furthermore, cellular proliferation assay was shown to be directly associated with miR-1301 levels. CONCLUSION: As a result, it was demonstrated that the over-expression of miR-1301 induced by the disruption of the KLF6 super enhancer leads to a significant inhibition of proliferation in HepG2 cells. Moreover, it was demonstrated that the KLF6 super enhancer regulates the cell-proliferative effects which are mediated, at least in part, by the induction of p21and p53 in a p53-dependent manner. Our results provide the functional significance of miR-1301 in understanding the transcriptional regulation mechanism of the KLF6 super enhancer.

An intronic mutation c.6430-3C>G in the F8 gene causes splicing efficiency and premature termination in hemophilia A.

: Hemophilia A is a bleeding disorder caused by coagulation factor VIII protein deficiency or dysfunction, which is classified into severe, moderate, and mild according to factor clotting activity. An overwhelming majority of missense and nonsense mutations occur in exons of F8 gene, whereas mutations in introns can also be pathogenic. This study aimed to investigate the effect of an intronic mutation, c.6430-3C>G (IVS22-3C>G), on pre-mRNA splicing of the F8 gene. We applied DNA and cDNA sequencing in a Chinese boy with hemophilia A to search if any pathogenic mutation in the F8 gene. Functional analysis was performed to investigate the effect of an intronic mutation at the transcriptional level. Human Splicing Finder and PyMol were also used to predict its effect. We found the mutation c.6430-3C>G (IVS22-3C>G) in the F8 gene in the affected boy, with his mother being a carrier. cDNA from the mother and pSPL3 splicing assay showed that the mutation IVS22-3C>G results in a two-nucleotide AG inclusion at the 3' end of intron 22 and leads to a truncated coagulation factor VIII protein, with partial loss of the C1 domain and complete loss of the C2 domain. The in-silico tool predicted that the mutation induces altered pre-mRNA splicing by using a cryptic acceptor site in intron 22. The IVS22-3C>G mutation was confirmed to affect pre-mRNA splicing and produce a truncated protein, which reduces the stability of binding between the F8 protein and von Willebrand factor carrier protein due to the loss of an interaction domain.

Terrimonas crocea sp. nov., isolated from the till of a high Arctic glacier.

A yellow-pigmented bacterial strain, designated M1-33108T, was isolated from the till of high Arctic glacier Midtre Lovénbreen near Ny-Ålesund, in the West Svalbard Archipelago, Norway. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain M1-33108T belonged to the genus Terrimonas and its closest neighbour was Terrimonas arctica R9-86T with 96.12 % 16S rRNA gene sequence similarity. Cells of strain M1-33108T were Gram-reaction-negative, aerobic, non-spore-forming, rod-shaped bacteria that lacked motility. Cells contained iso-C15 : 1 G, iso-C15 : 0, iso-C17 : 0 3-OH, C16 : 0 and summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c) as its major cellular fatty acids and menaquinone-7 as the sole respiratory quinone. The polar lipid profile of strain M1-33108T consisted of phosphatidylethanolamine, two unknown aminophospholipids, eight unknown aminolipids, an unknown glycolipid and three unknown polar lipids. The DNA G+C content was 45.0 mol%. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain M1-33108T is considered to represent a novel species in the genus Terrimonas, for which the name Terrimonas crocea sp. nov. is proposed. The type strain is M1-33108T (=CCTCC AB 2016103T=KCTC 52448T).

Hymenobacter rutilus sp. nov., isolated from marine sediment in the Arctic.

Strain K2-33028T, which appeared as a brick-red colony on an R2A plate, was isolated from a marine sediment sample from Kings Bay, Svalbard Archipelago, Norway. Phylogenetic analysis based on 16S rRNA gene sequences indicated that K2-33028T represented a member of the genus Hymenobacter. Cells were Gram-reaction-negative, non-spore-forming, aerobic, rod-shaped and without motility. Growth occurred at 4-37 °C (optimum 28 °C) and at pH 6.0-8.0 (optimum pH 7.0). Cells contained menaquinone-7 as the main respiratory quinone and iso-C15 : 0, summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c), C16 : 1ω5c, summed feature 4 (comprising anteiso-C17 : 1B and/or iso-C17 : 1I) and anteiso-C15 : 0 as the major cellular fatty acids. Phosphatidylethanolamine was predominant in the polar lipid profile. The DNA G+C content was 64.3 mol%. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain K2-33028T is considered to represent a novel species of the genus Hymenobacter, for which the name Hymenobacterrutilussp. nov. is proposed. The type strain is K2-33028T (=CCTCC AB 2016091T=KCTC 52447T).

Study on the vibration displacement distribution of a circular ultrasonic motor stator.

In this paper is presented a theoretical consideration on the stator's displacement distribution, which is one of the most important problems in defining the structure of a circular ultrasonic motor stator. The results are compared with ones obtained utilizing a holographic interferometer, a laser vibrometer and FEM (finite element method) simulation. They are in good agreement with each other.