Development of an in vitro screening system for synthetic signal peptide in mammalian cell-based protein production.

Optimizing appropriate signal peptides in mammalian cell-based protein production is crucial given that most recombinant proteins produced in mammalian cells are thought to be secreted proteins. Until now, most studies on signal peptide in mammalian cells have replaced native signal peptides with well-known heterologous signal peptides and bioinformatics-based signal peptides. In the present study, we successfully established an in vitro screening system for synthetic signal peptide in CHO cells by combining a degenerate codon-based oligonucleotides library, a site-specific integration system, and a FACS-based antibody detection assay. Three new signal peptides were screened using this new screening system, confirming to have structural properties as signal peptides by the SignalP web server, a neural network-based algorithm that quantifies the signal peptide-ness of amino acid sequences. The novel signal peptides selected in this study increased Fc-fusion protein production in CHO cells by increasing specific protein productivity, whereas they did not negatively affect cell growth. Particularly, the SP-#149 clone showed the highest qp, 0.73 ± 0.01 pg/cell/day from day 1 to day 4, representing a 1.47-fold increase over the native signal peptide in a serum-free suspension culture mode. In addition, replacing native signal peptide with the novel signal peptides did not significantly affect sialylated N-glycan formation, N-terminal cleavage pattern, and biological function of Fc-fusion protein produced in CHO cells. The overall results indicate the utility of a novel in vitro screening system for synthetic signal peptide for mammalian cell-based protein production. KEY POINTS: • An in vitro screening system for synthetic signal peptide in mammalian cells was established • This system combined a degenerate codon-based library, site-specific integration, and a FACS-based detection assay • The novel signal peptides selected in this study could increase Fc-fusion protein production in mammalian cells.

Multiple Daily Rounds of Theta-Burst Stimulation for Tinnitus: Preliminary Results.

Background and Objectives: Tinnitus is a condition that negatively affects the quality of life and is difficult to treat. Theta burst stimulation (TBS), a new method of repetitive transcranial magnetic stimulation (rTMS), is a promising treatment approach because it shows stronger and more prolonged effects in a shorter time of stimulation than other rTMS protocols. However, the therapeutic effect of TBS for tinnitus was inconsistent. We hypothesized that more stimulation would be more effective. Therefore, this study aimed to explore the safety and effectiveness of multiple daily rounds of TBS over five consecutive days. Materials and Methods: The continuous TBS (cTBS) protocol is 300 pulses/day, but we applied 8 sessions of 300 pulses in a day (total 2400 pulses/day). A total of 15 patients with tinnitus were randomly assigned to treatment and sham groups. Outcome measurements were taken three times: before and after 5-day of stimulation; at a 1-3 month follow-up visit. Outcome measurements were the degree of annoyance due to ear fullness, duration of tinnitus, visual analog scales of tinnitus for annoyance, Tinnitus Handicap Inventory, pitch, loudness, minimum masking level, and residual inhibition. Results: Five-day cTBS was completed without adverse events. We did not find any significant therapeutic effect in the treatment group, but we needed to be cautious to interpret our result due to the small sample size. Conclusions: In conclusion, multiple rounds of cTBS in a day may be safe. Further research is needed in a larger sample size to determine the effectiveness and confirm the safety.

Selective Generation of Lamb Wave Modes in a Finite-Width Plate by Angle-Beam Excitation Method.

A Lamb wave in a plate with a finite width has both thickness and width modes, whereas only thickness modes exist in an infinitely wide plate. The thickness and width modes are numerously formed in a finite-width plate, and they all have different cut-off frequencies, wave velocities, and wave structures. These different characteristics can be utilized in various applications, but a selective generation method for a particular Lamb wave mode in a finite-width plate has not been sufficiently studied, and only a method using multiple elements has been reported. This paper presents the selective generation of a certain Lamb wave mode in a finite-width plate by an angle-beam excitation method using single or dual wedges. In the proposed generation method, a specially designed wedge with grooves or a patch having insulation layers is employed for partial acoustic insulation of the ultrasonic energy incident into the plate. The feasibility of the proposed method was investigated through finite element method (FEM) simulations for Lamb wave excitation and propagation, and then experimentally demonstrated by the measurement of Lamb wave propagation using a laser scanning vibrometer.

Ear, Nose, and Throat Foreign Bodies in Children: A Retrospective Study.

BACKGROUND: This study analyzed the presentation, characteristics, and management of foreign bodies in different age groups of pediatric patients with ear, nose, and throat foreign bodies. METHODS: A retrospective study was performed using data from October 2012 to September 2020. A total of 1285 patients with ear, nose, and throat foreign bodies who were less than 12 years of age and who presented to the emergency room were included in this study. Their biographical data, clinical presentations, foreign body types and locations, and management outcomes were obtained from medical records and analyzed as three age groups (infancy: <2 years old; early childhood: 2-5 years old; and late childhood: 6-12 years old). RESULTS: The early childhood group had the highest number of patients (n = 672; 52.2%). Throat was the most common location (59.2%), and bone was the most common type of foreign body. Among the children who visited our hospital, foreign bodies were actually found in only 657 patients (51.1%) and removed by an otolaryngologist in 625 (95.1%) cases. CONCLUSION: Our study could provide guidance for the diagnosis and management of pediatric patients who present to emergency departments with foreign bodies.

Association between Thyroid Cancer and Weight Change: A Longitudinal Follow-Up Study.

BACKGROUND: The purpose of this study was to evaluate body mass index (BMI) and systolic blood pressure (SBP)/diastolic blood pressure (DBP) between Korean adults who underwent thyroidectomy and comparison groups. METHODS: Data were included from the Korean National Health Insurance Service-Health Screening Cohort (2002-2015). BMI and SBP/DBP were measured before thyroidectomy, 1 and 2 years after thyroidectomy (n = 1995 in study I, n = 2162 in study II), comparing 1:4 matched participants (n = 7980 in study I, n = 8648 in study II). The paired t-test and linear mixed model were used to identify the differences between groups. RESULTS: DBP in both thyroid cancer II and comparison II group were significantly lower after thyroidectomy than before thyroidectomy. However, the interaction effect of thyroidectomy in study I and study II did not reach statistical significance. CONCLUSION: BMI, SBP and DBP were not significantly different between the thyroidectomy group and the matched comparison group.

Effect of noise and reverberation on speech recognition and listening effort for older adults.

AIM: The purpose of the present study was to evaluate the speech recognition ability and degree of listening effort for older adults in noisy and reverberating environments. METHODS: A total of 80 participants (40 older and 40 younger adults) participated in a sentence recognition test under 20 conditions with four levels of background noise and five levels of reverberation time. In each condition, the degree of listening effort was self-reported using a 5-point Likert scale. RESULTS: The older and younger groups showed that both the error percentage of the sentence recognition task and scale of the listening effort increased as the background noise and reverberation time increased. In the sentence recognition task, the older group was more affected by the reverberation condition than the noisy background compared with their younger counterparts. For the listening effort, the older group had higher scales than the younger group in all conditions, thereby explaining that they required significant effort during the listening task. CONCLUSIONS: The older adults had poorer speech perception ability and required more listening effort than their younger counterparts in all conditions. Reverberation reversely significantly affected the speech recognition and listening effort for the older adults when compared with the background noise, suggesting that an appropriate level of noise and reverberation should be considered for comfort and a less stressful listening environment for the older adult population. Geriatr Gerontol Int 2018; 18: 1603-1608.

Korean herbal medicine for treating henoch-schonlein purpura with yin deficiency: five case reports.

OBJECTIVES: The purpose of this study is to report the clinical effect of Korean medicine (KM) treatment for Henoch-Schonlein purpura (HSP). METHODS: Five HSP patients who demonstrated a Yin deficiency and who had a history of a previous upper respiratory tract infection were included in this study. Four patients had arthritis and three had severe stomachache. One of them appeared to have proteinuria and hematuria before starting KM treatment. RESULTS: All patients were improved with only herbal medicine, Jarotang (JRT). Purpura in the lower extremities and abdominal pain, which were not treated by using a corticosteroid, disappeared and had not recurred after 6 months. CONCLUSION: These cases indicate that JRT may be effective in treating HSP in patients who demonstrate Yin deficiency, even though the number of cases was limited to five.

The therapeutic effect of neurologic music therapy and speech language therapy in post-stroke aphasic patients.

OBJECTIVE: To investigate the therapeutic effect of neurologic music therapy (NMT) and speech language therapy (SLT) through improvement of the aphasia quotient (AQ) in post-stroke aphasic patients. METHODS: Twenty-one post-stroke, nonfluent aphasia patients who had ischemic/hemorrhagic stroke on radiologic evaluation were divided into the NMT and SLT groups. They received NMT and SLT for 1 month. Language function was assessed by Korean version-Western Aphasia Battery before and after therapy. NMT consisted of therapeutic singing and melodic intonation therapy, and SLT consisted of language-oriented therapy. RESULTS: Significant improvements were revealed in AQ, repetition, and naming after therapy in the NMT group and improvements in repetition in the SLT group of chronic stroke patients (p<0.05). There were significant improvements in language ability in the NMT group of subacute stroke patients. However, there was no significant improvement in the SLT group of subacute stroke patients. CONCLUSION: We concluded that the two therapies are effective treatments in the chronic stage of stroke and NMT is effective in subacute post-stroke aphasic patients.

In vitro and in vivo mechanical stability of orthodontic mini-implants.

OBJECTIVE: To compare in vivo and in vitro mechanical stability of orthodontic mini-implants (OMIs) treated with a sandblasted, large-grit, and anodic-oxidation (SLAO) method vs those treated with a sandblasted, large-grit, and acid-etching (SLA) method. MATERIALS AND METHODS: Fifty-four titanium OMIs (cylindrical shape, drill-free type; diameter  =  1.45 mm, length  =  8 mm, Biomaterials Korea Inc, Seoul, Korea) were allocated into control, SLA, and SLAO groups (N  =  12 for in vivo and N  =  6 for in vitro studies per group). In vitro study was carried out on a polyurethane foam bone block (Sawbones, Pacific Research Laboratories Inc, Vashon, Wash). In vivo study was performed in the tibias of Beagles (6 males, age  =  1 year, weight  =  10 to 13 kg; OMIs were removed at 8 weeks after installation). For insertion and removal of OMIs, the speed and maximum torque of the surgical engine were set to 30 rpm and 40 Ncm, respectively. Maximum torque (MT), total energy (TE), and near peak energy (NPE) during the insertion and removal procedures were statistically analyzed. RESULTS: In the in vitro study, although the control group had a higher insertion MT value than the SLA and SLAO groups (P < .01), no differences in insertion TE and NPE or in any of the removal variables were noted among the three groups. In the in vivo study, the control group exhibited higher values for all insertion variables compared with the SLA and SLAO groups (MT, P < .001; TE, P < .01; NPE, P < .001). Although no difference in removal TE and removal NPE was noted among the three groups, the SLAO group presented with a higher removal MT than the SLA and control groups (P < .001). CONCLUSIONS: SLAO treatment may be an effective tool in reducing insertion damage to surrounding tissue and improving the mechanical stability of OMIs.

Unsteady flow characteristics through a human nasal airway.

Time-dependent characteristics of the flow in a human nasal airway constructed from the CT image of a healthy volunteer were investigated using a computational fluid dynamics (CFD) technique. To capture the time-varying nature of the flow as well as pressure and temperature fields, the large eddy simulation (LES) technique instead of the RANS (Reynolds Averaged Navier-Stokes) approach was adopted. To make the present analysis more relevant to a real human breathing cycle, the flow was designed to be induced by the pressure difference and the time-varying pressure at the end of trachea was described to reproduce the flow rate data from the measurement. Comparison of the present results with those of typical steady simulations showed that the difference in flow characteristics is magnified in the expiration phase. This fact may suggest that the inertial effect associated with unsteady flow is more important during the expiration period. Also, the fact that the distribution of the flow rate in a given cross-section of the airway changes significantly with time implies the importance of unsteady data for clinical decision. The wall shear stress was found to have relatively high values at the locations near nasopharynx and larynx but the magnitude changes with time during the whole respiratory cycle. Analysis of the temperature field showed that most of the temperature change occurs in the nasal cavity when the air is incoming and thus, the nasal cavity acts as a very efficient heat exchanger during an inspiration period.

A new nose tip-defining technique for Asians using autologous cartilage prefabricated with octyl-2-cyanoacrylate adhesive.

OBJECTIVES: To form and test a prefabricated, 3-dimensional tip graft structure from autologous cartilage pieces and octyl-2-cyanoacrylate adhesive (Dermabond). Displacement and instability of grafted cartilage are common drawbacks of endonasal tip surgery, and we evaluated the feasibility and efficacy of octyl-2-cyanoacrylate adhesive for clinical use. METHODS: Three New Zealand rabbits were examined in this study. We cut auricular cartilage into pieces of approximately 1 x 2 cm and glued them together with octyl-2-cyanoacrylate. Three months after subcutaneous implantation in the rabbits, we histopathologically analyzed foreign body reactions, histotoxic effects, and the structure of the regenerative tissue in the implanted cartilage. Furthermore, from human patients we harvested small pieces of autologous cartilage and cemented them together with octyl-2-cyanoacrylate adhesive to prefabricate 3-dimensional tip grafts, which we then inserted into the nasal tips of patients via an endonasal approach to project and contour the nasal tip. The changes in tip definition of 42 Asian patients were analyzed. RESULTS: Octyl-2-cyanoacrylate adhesive interacts well with cartilage tissues with no systemic or local adverse effects. Its ability to bond cartilage instantaneously allows for the fabrication of intricate cartilage implant components. The aesthetic results in 42 cases were excellent, and the implants were both safe and effective. CONCLUSION: Our results demonstrate that octyl-2-cyanoacrylate can be safely used in endonasal tip surgery to produce excellent cosmetic outcomes in Asian subjects.

Simple quantitative analysis of Escherichia coli K-12 internalized in baby spinach using Fourier Transform Infrared spectroscopy.

Bacterial contamination continues to be a serious concern for food safety. Although washing fresh produce helps in reducing pathogen levels, pathogen internalization often limits the effectiveness of washing. When pathogens internalize in leafy vegetables, the method of identification and quantitative measurement would be called into question. This study was aimed to use Fourier Transform Infrared (FTIR) spectroscopy integrated with an attenuated total reflectance kit for quantification of Escherichia coli K-12 internalized in baby spinach. The bacteria were inoculated into vascular and intracellar tissues of spinach leaves by syringe injection and the distribution of internalized E. coli K-12 cells was confirmed under scanning electron microscopy (SEM). FTIR measurement following the preparation of bacterial suspension from spinach leaves with high speed pulverizing enabled to detect the absorbance peaks in the amide II region between 1590 and 1490 cm⁻¹ as a fingerprint for the microbes. It was found that the estimated concentrations of E. coli K-12 agreed well with the concentrations determined by plate counting with R² values of 0.98 and 0.97 in peptone water and spinach extracts, respectively. The results demonstrated that FTIR can identify and quantify E. coli K-12 in baby spinach extracts at a limit of detection of approximately 100 CFU/mL in 5 min. The developed method is expected to be suitable for the analysis of pathogenic E. coli strains and other bacterial species in fresh vegetables.

The science of ecological economics: a content analysis of Ecological Economics, 1989-2004.

The Ecological Economics journal is a primary source for inquiry on ecological economics and sustainability. To explore the scholarly pursuit of ecological economics, we conducted a content analysis of 200 randomly sampled research, survey, and methodological articles published in Ecological Economics during the 15-year period of 1989-2004. Results of the analysis were used to investigate facets of transdisciplinarity within the journal. A robust qualitative approach was used to gather and examine data to identify themes representing substantive content found within the span of sampled journal papers. The extent to which each theme was represented was counted as well as additional data, such as author discipline, year published, etc. Four main categories were revealed: (1) foundations (self-reflexive themes stemming from direct discussions about ecological economics); (2) human systems, represented by the themes of values, social indicators of well-being, intergenerational distribution, and equity; (3) biophysical systems, including themes, such as carrying capacity and scarcity, energy, and resource use, relating directly to the biophysical aspects of systems; and (4) policy and management encompassing themes of development, growth, trade, accounting, and valuation, as well as institutional structures and management. The results provide empirical evidence for discussing the future direction of ecological economic efforts.

Nasal airflow during respiratory cycle.

BACKGROUND: Knowledge on the airflow patterns in the nasal cavity is essential to understanding the function of the nasal cavity. This study has attempted to observe the breath cycle of nasal airflow during respiration at rest. METHODS: We constructed a nasal cavity model by rapid prototyping using 1.25-mm-thick CT data and devised a piston pump driven by a cam, to simulate respiration at rest. The airflow was evaluated with particle image velocimetry and visualized in coronal reconstructed images. RESULTS: During the inspiration, a maximal velocity was observed at the valve area and the main stream occurred in the middle and superior airways. During the expiration, main stream was noted in the middle airway and was slow compared with the flow during inspiration. Vortexes were observed between inspiration and expiration. CONCLUSION: This result widens our knowledge of nasal airflow and this technique will allow a more physiological understanding of nasal operations.