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This study investigates a method for improving real-time decisions regarding the storage location of export containers while the containers are arriving. To manage the decision-making process, we propose a two module-based data-driven dynamic stacking strategy that facilitates stowage planning. Module 1 generates the Gaussian mixture model (GMM) specific to each container group for container weight classification. Module 2 implements the data-driven dynamic stacking strategy as an online algorithm to determine the storage location of an arriving container in real time. Numerical experiments were conducted using real-life data to validate the effectiveness of the proposed method compared to other alternative stacking strategies. These experiments revealed that the performance of the proposed method is robust, and therefore it can improve yard operations and container terminal competitiveness.

AIM To investigate the difference in the initial surgical results between a new monofocal intraocular lens (IOL) with enhanced intermediate vision and the standard monofocal IOL in patients with retinal disease. METHODS We retrospectively reviewed the medical records of patients with retinal disease who underwent cataract surgery due to accompanying cataracts. Preoperative best corrected distant visual acuity, spherical equivalent(SE), and types of retinal diseases were investigated. The surgical results were investigated using uncorrected visual acuity(UCVA) and SE at 1 day, 1 week, and 1 month after surgery. RESUTLS Seventeen eyes treated with a new monofocal IOL enhanced for intermediate vision (ICB00 group) and 18 eyes treated with the standard monofocal IOL (AAB00 group) were included in this study. There were no significant differences in the baseline characteristics, including the type of underlying retinal disease, between the groups. There were no significant differences between the groups in terms of distant, intermediate, or near UCVA at day 1 and week 1 after surgery. However, at 1 month after surgery, the ICB00 group showed a significantly better intermediate vision improvement than the AAB00 group (p = 0.001). CONCLUSION Even in patients with cataract accompanied by retinal disease, the use of the ICB00 IOL showed significant improvement in intermediate vision compared to the use of the AAB00 (standard monofocal) IOL. The ICB00 IOL might be a good option for patients with cataract and retinal disease in the era of increased intermediate vision needs in daily life.

PURPOSE To evaluate the clinical efficacy and safety of immediate manual meibomian gland expression (MGX) after LipiFlow thermal pulsation (TearScience Inc, Morrisville, NC) for obstructive meibomian gland dysfunction and to compare the LipiFlow only and MGX after LipiFlow. METHODS Patients who underwent immediate manual MGX after LipiFlow or who received only LipiFlow treatment were included. Thirty eyes from 15 patients were enrolled in each group. All patients underwent 3 treatments at monthly intervals. All patients were followed up for 6 months after treatment. All patients were examined before and at 3 and 6 months after treatment. Examinations included the Ocular Surface Disease Index score, noninvasive tear film breakup time (NIBUT), lipid layer thickness (LLT), corneal and conjunctival staining, and tear meniscus height. RESULTS The Ocular Surface Disease Index scores improved in both groups during the follow-up periods (P = 0.001 and P = 0.001). In the LipiFlow-only group, the NIBUT and LLT significantly improved at 3 months (P < 0.001 and P = 0.006) but deteriorated at 6 months. In the MGX after LipiFlow group, the NIBUT and LLT improved at 3 months (P < 0.001 and P < 0.001), and this improvement was maintained at 6 months. The improvement of NIBUT at 3 months was greater in the MGX after LipiFlow group (3.24 ± 1.16 to 9.25 ± 1.36 s) than in the LipiFlow-only group (3.78 ± 1.75 to 7.18 ± 2.70 s), and the improvements of the LLT at 6 months were greater in the MGX after LipiFlow group (30.27 ± 10.74 to 46.93 ± 20.81 μm) than in the LipiFlow-only group (34.70 ± 10.79 to 38.73 ± 14.70 μm). CONCLUSIONS Both LipiFlow only and MGX after LipiFlow were clinically effective for obstructive meibomian gland dysfunction. However, the efficacy and persistence of treatment were greater in patients who received MGX after LipiFlow.

AIM: To evaluate clinical outcomes of unilateral implantation of a diffractive multifocal intraocular lens (IOL) in patients with contralateral monofocal IOL. METHODS: Twenty-two patients who already had implantation of a monofocal IOL in unilateral eye underwent implantation of a diffractive multifocal IOL in contralateral eye were enrolled. After 1, 6, and 12mo, uncorrected and distant corrected distant visual acuity (UCDVA and DCDVA), uncorrected and distant corrected intermediate-visual acuity (UCIVA and DCIVA), uncorrected and distant corrected near visual acuity (UCNVA and DCNVA), and contrast sensitivity were obtained. Halo/glare symptoms, spectacle dependence, and patient satisfaction were also evaluated. RESULTS: The mean age was 67.86±7.25y and the average interval between two IOL implantations was 645.82±878.44d. At 1mo, binocular UCDVA was lower than 0.20 logMAR in 76% of patients (mean 0.12±0.13 logMAR), which increased to 90% by 6 and 12mo. The binocular UCDVA was significantly better than the monocular results (P

Purpose To compare the protective effects of cyclosporine A emulsion (Restasis: 0.05% cyclosporine A) (CsAE) and cyclosporine A cationic emulsion (Ikervis: 0.1% cyclosporine A) (CsACE) on cellular inflammation, apoptosis, proliferation, and survival in an in vitro dry eye model. Methods The concentration of CsA in CsAE and CsACE was verified using a liquid chromatography tandem mass spectrometry system. Human corneal epithelial cells were subjected to desiccation stress. Human corneal epithelial cells were incubated with or without 3 groups of cyclosporine A medium (CsAE 1:50, CsACE 1:50, and CsACE 1:100). p-NF-κB p65, p-IκBα, Bax, Bcl-xL, p-Erk1/2, and p-Akt levels were determined using Western blots, and TNF-α levels were quantified using an enzyme-linked immunosorbent assay. Results The CsA concentration of CsACE 1:100 was nearly the same as that of CsAE 1:50. Compared with CsAE 1:50 (0.78 ± 0.19 fold), the p-NF-κB p65 level was further reduced in CsACE 1:50 (0.38 ± 1.20 fold) and 1:100 (0.29 ± 0.11 fold) as well as p-IκBα. Levels of TNF-α were also lower in CsACE 1:50 and 1:100 than in CsAE 1:50. Induction of the apoptotic protein Bax was significantly decreased in CsACE 1:50 and 1:100 compared with CsAE 1:50, whereas that of the antiapoptotic protein Bcl-xL was increased in CsACE 1:50 and 1:100. p-ERK1/2 and p-Akt levels were higher in CsACE 1:50 and 1:100 than in CsAE 1:50. Conclusions CsACE had more potent anti-inflammatory and antiapoptotic effects than CsAE in a transwell desiccation stress model. CsACE also enhanced proliferation and survival factors under desiccation stress compared with CsAE in this in vitro dry eye model.

Wind and solar energy are recognized as environmentally friendly energies. Wind energy is generated from the surface of the earth by solar energy. While wind energy can be regarded as "free" energy, generating it entails high installation cost as well as a large land footprint and noise, which is deemed to be a nuisance. In addition, maintenance costs are high. In this study, we propose a novel tree-wrapped wind energy harvester, which is inexpensive and easy to manufacture. Triboelectric generators follow simple principles and can be of various generator types. Triboelectric generators running on wind energy have been studied by many researchers. Our design comprises light and flexible tree-wrapped triboelectric generator modules. Multiple generator modules can be connected in parallel to harvest electrical energy. We show that the proposed generator can potentially harvest sufficient wind energy and can even be used in an urban environment by mounting them on trees in cities.

In this study, we propose a module-type triboelectric nanogenerator (TENG) capable of harvesting electricity from a variety of mechanical energy sources and generating power from diverse forms that fit the modular structure of the generator. The potential energy and kinetic energy of water are used for the rotational motion of the generator module, and electricity is generated by the contact/separation generation mode between the two triboelectric surfaces inside the rotating TENG. Through the parametric design of the internal friction surface structure and mass ball, we optimized the output of the proposed structure. To magnify the power, experiments were conducted to optimize the electrical output of the series of the TENG units. Consequently, outputs of 250 V and 11 μA were obtained when the angle formed between the floor and the housing was set at 0° while nitrile was set as the positively charged material and the frequency was set at 7 Hz. The electrical signal generated by the module-type TENG can be used as a sensor to recognize the strength and direction of various physical quantities, such as wind and earthquake vibrations.

To evaluate the clinical efficacy and safety of combination treatment with intense pulsed light (IPL), LipiFlow thermal pulsation, and manual meibomian gland expression (MGX) for patients with refractory meibomian gland dysfunction (MGD).A total of 23 eyes from 23 patients who underwent combination treatment with IPL, LipiFlow, and MGX for refractory MGD were included in the study. All patients were followed up for 12 months after the first treatment. All patients were examined prior to treatment and underwent three sessions of IPL, LipiFlow, and MGX, a subsequent three sessions of IPL and MGX, and six subsequent sessions of MGX. Patients were examined 10 weeks, 6 months, and 12 months after the first treatment. The ophthalmologic examination included ocular surface disease index (OSDI), non-invasive tearfilm break-up time (NIBUT), lipid layer thickness (LLT), meibomian gland expressibility (MGE), meibomian gland morphology (meiboscore), and tear meniscus height (TMH).OSDI, NIBUT, LLT, and MGE significantly improved with treatment, which was maintained during the follow-up period. TMH and meiboscore remained unchanged throughout follow-up. No serious adverse events occurred during follow-up.Combination therapy with IPL, LipiFlow, and MGX was clinically effective and safe for the treatment of refractory MGD. Combination treatment could be considered for patients with insufficient improvement despite conventional MGD treatments, including IPL treatment with MGX.

Highly flexible and compressible porous polyurethane (PU) structures have effectively been applied in capacitive pressure sensors because of the good elastic properties of the PU structures. However, PU porous structure-based pressure sensors have been limited in practical applications owing to their low durability during pressure cycling. Herein, we report a flexible pressure sensor based on a three-dimensional porous structure with notable durability at a compressive pressure of 500 kPa facilitated by the use of a shape memory polymer (SMP). The SMP porous structure was fabricated using a sugar templating process and capillary effect. The use of the SMP resulted in the maintenance of the sensing performance for 100 cycles at 500 kPa, the SMP can restore its original shape within 30 s of heating at 80 °C. The pressure sensor based on the SMP exhibited a higher sensitivity of 0.0223 kPa−1 than a typical PU-based sensor and displayed excellent sensing performance in terms of stability, response time, and hysteresis. Additionally, the proposed sensor was used to detect shoe insole pressures in real time and exhibited remarkable durability and motion differentiation.

Maritime Informatics can support recreational boating and professional fishing vessels in communicating with relevant actors in their environment. There are opportunities for digital solutions for recreational and fishing vessels, when they use the digital infrastructures developed for merchant shipping. When ships and vessels use the same waterways, they should all have access to the same digital infrastructure and services to enable complete situational awareness and thus improve safety, environmental sustainability and efficiency for all at sea.

In recent times, polymer-based flexible pressure sensors have been attracting a lot of attention because of their various applications. A highly sensitive and flexible sensor is suggested, capable of being attached to the human body, based on a three-dimensional dielectric elastomeric structure of polydimethylsiloxane (PDMS) and microsphere composite. This sensor has maximal porosity due to macropores created by sacrificial layer grains and micropores generated by microspheres pre-mixed with PDMS, allowing it to operate at a wider pressure range (~150 kPa) while maintaining a sensitivity (of 0.124 kPa&minus, 1 in a range of 0~ 15 kPa) better than in previous studies. The maximized pores can cause deformation in the structure, allowing for the detection of small changes in pressure. In addition to exhibiting a fast rise time (~167 ms) and fall time (~117 ms), as well as excellent reproducibility, the fabricated pressure sensor exhibits reliability in its response to repeated mechanical stimuli (2.5 kPa, 1,000 cycles). As an application, we develop a wearable device for monitoring repeated tiny motions, such as the pulse on the human neck and swallowing at the Adam&rsquo, s apple. This sensory device is also used to detect movements in the index finger and to monitor an insole system in real-time.

AIM: To investigate the ocular biodistribution and clearance of topically administered 7-taurocholic acid conjugated low-molecular weight heparin (LHT7) in a neovascularized mouse cornea using an in vivo optical imaging system. METHODS: A total of 10 eyes of 6 to 8-week-old BALB/c mice were analyzed. Corneal neovascularization (CoNV) was induced in the inferior cornea (IC) of each animal by penetrating the stroma with two interrupted sutures. The development of CoNV was verified after one week and the area of each neovascularized region was measured. A near-infrared fluorescent probe of 20 µmol/L Cy5.5 labeled LHT7 (LHT7-Cy5.5) in 0.02 mL solution was topically instilled onto the cornea in the experimental group (n=5). Free-Cy5.5 of 20 µmol/L in 0.02 mL was instilled in the control group (n=5). In vivo optical images were obtained before instillation and 5min, 2, 4, and 6h after instillation. The intensities were separately measured at the superior cornea (SC) and the IC. RESULTS: The mean CoNV areas were 1.97±0.17 mm2 and 1.92±0.96 mm2 in the experimental and control groups, respectively (P=0.832). The SC remained normal in all 10 subject animals. The IC intensity of the LHT7-Cy5.5 was greater than the SC intensity at 5min (P=0.038), 2h (P=0.041), and 4h (P=0.041) after application. The IC intensity fell to less than half of its initial value (42.9%±8.6%) at 6h in the experimental group. In the control mice, here were no significant differences in the free-Cy5.5 intensity between the IC and SC. CONCLUSION: Topically administered LHT7 shows a high biodistribution in CoNV areas for 4h and should be reapplied accordingly to maintain its effects. In vivo optical imaging can be a useful tool for evaluating the ocular biodistribution of a drug in an animal model.

Thick biological tissues give rise to not only the multiple scattering of incoming light waves, but also the aberrations of remaining signal waves. The challenge for existing optical microscopy methods to overcome both problems simultaneously has limited sub-micron spatial resolution imaging to shallow depths. Here we present an optical coherence imaging method that can identify aberrations of waves incident to and reflected from the samples separately, and eliminate such aberrations even in the presence of multiple light scattering. The proposed method records the time-gated complex-field maps of backscattered waves over various illumination channels, and performs a closed-loop optimization of signal waves for both forward and phase-conjugation processes. We demonstrated the enhancement of the Strehl ratio by more than 500 times, an order of magnitude or more improvement over conventional adaptive optics, and achieved a spatial resolution of 600 nm up to an imaging depth of seven scattering mean free paths., Optical imaging deep in biological tissue is difficult due to multiple scattering and specimen induced aberrations of both the incident and reflected light. Here, Kang et al. develop an adaptive closed-loop algorithm to correct tissue aberrations in the presence of multiple scattering for deep tissue imaging.

Chinese hamster ovary (CHO) cells are the most widely used host cell lines for the commercial production of therapeutic proteins including Fc-fusion proteins. During the culture of recombinant CHO (rCHO) cells, host cell proteins (HCPs), secreted from viable cells and released from dead cells, accumulate extracellularly, potentially impairing product quality. In this study, the HCPs that accumulated extracellularly in batch and fed-batch cultures of Fc-fusion protein-producing rCHO cell lines (DG-Fc and DUKX-Fc) were identified and quantified using nanoflow liquid chromatography-tandem mass spectrometry (LC-MS/MS), followed by gene ontology and functional analysis. When the proteome database of Cricetulus griseus was used as a reference to identify the HCPs, more HCPs were identified for DG-Fc (1632 HCPs in batch culture and 1733 HCPs in fed-batch culture) than for DUKX-Fc (1114 HCPs in batch culture and 1002 HCPs in fed-batch culture). Clustering analysis of HCPs, which were classified into four clusters according to their concentration profiles during culture, showed that the concentration profiles of HCPs affecting the quality of Fc-fusion proteins correlated with changes in Fc-fusion protein quality. Taken together, the dataset of HCPs obtained in this study using the two different rCHO cell lines provides insights into the determination of appropriate target proteins to be removed during the culture and purification steps so as to ensure good Fc-fusion protein quality. Biotechnol. Bioeng. 2017;114: 2267-2278. © 2017 Wiley Periodicals, Inc.

The mechanical properties of copper foils electroplated with (200) and (220) preferred orientations and thicknesses of 8–24 μm were assessed at various temperatures. It was found that the yield strength and elastic modulus both decrease with increasing temperature to 150 °C, and that the greater susceptibility of a columnar structure to defects makes it more sensitive to temperature than an equiaxed structure. Thus, for a given grain size, a columnar structure has a lower yield strength. A modified Hall-Petch relation was subsequently developed to take into account the effect of micro-structure on material strength by introducing a structure factor (S) : σy = σ0 + kd− 1/2 + S. As the slope (k) of this equation is constant, it can be used for copper thin films of any thickness, provided they are at room temperature and above the micro-level in scale. This structure factor concept, however, is only valid at room temperature. In the modified Hall-Petch relation, the new variable related to the equiaxed and columnar structures in the copper thin films was abstracted from the Hall-Petch relation. Then, if the materials are the same, the k value and the σ0 value are constant regardless of the scale, and the relationship between the grain size and the yield strength can be shown by only changing the structure factor.

Purpose To evaluate and compare the predictability and safety of immediate small-incision lenticule extraction, also known as SMILE, after suction loss during the procedure with those of uneventful small-incision lenticule extraction. Setting Nunemiso Eye Center, Seoul, South Korea. Design Retrospective case series. Methods Patients who had immediate small-incision lenticule extraction using reapplied suction (study group) or uneventful small-incision lenticule extraction (control group) for myopia or myopic astigmatism were included. All patients were followed for at least 12 months. Demographic as well as visual and refractive outcomes were compared between the 2 groups. Results Twenty-three eyes were enrolled in the study group and 48 eyes in the control group. The overall cumulative incidence of suction loss was 0.2% (28 of 11 762 eyes). Twenty-two eyes (96%) in the study group had unchanged or improved decimal Snellen corrected distance visual acuity (CDVA). At 12 months, the mean uncorrected distance visual acuity, CDVA, and manifest refraction defocus equivalent in the study group and control group were 0.99 ± 0.17 (SD), 1.11 ± 0.14, 0.75 ± 0.50 diopters (D), and 1.25 ± 0.22 D, 1.26 ± 0.21 D, 0.35 ± 0.26 D, respectively (P = .000, P = .007, and P = .000, respectively). The correlation between the achieved and the attempted spherical equivalent refraction was slightly better in the control group (R2 = 0.972) than in the study group (R2 = 0.933). Conclusions Immediate small-incision lenticule extraction using reapplied suction resulted in safe and clinically predictable long-term outcomes. However, predictability was lower for immediate than for uneventful small-incision lenticule extraction.

Energy harvesting, especially for powering low-power internet-of-things (IoT) devices, is gaining attention in recent years. Triboelectric nanogenerators have been studied to improve the output by applying a structure that can concentrate electrons on the surface of the generator materials. For enhancing the triboelectrification output, we herein focused on the power output line. A method for increasing the amount of electrons on the power lead by potential difference and their acceleration was studied. A rod was shaken by external vibrations, the accumulated charges were discharged in a manner similar to that of a lightning rod. Micro plasma was generated when the rod made contact with the mating micro-cracked surface innumerable times. The micro-cracked surface was fabricated with a diamond tip moving horizontally to the surface. As the resistance of the micro plasma was close to zero, the amount of electron movement was instantaneously accelerated. This type of triboelectric generator can be fabricated in the form of an electric box. By using this triboelectric power amplifier, voltage can be amplified 2 to 3 times, and the current can be amplified 10 to 15 times, thus, enhanced energy harvesting efficiency is attained.

Study Objective How perioperative heart rate variability (HRV) indices differ according to the anxiety or depressed mood of patients scheduled to undergo a major surgical procedure for cancer. Design Prospective observational study. Setting Operating room. Patients Forty-one male patients between 40 and 70 years of age with hepatocellular carcinoma were included in the final analysis. Interventions HRV was measured on the day before surgery (T1), impending anesthesia (T2), and after anesthetic induction (T3). Preoperative anxiety and depressed mood of all patients were evaluated using the State-Trait Anxiety Inventory and Self-Rating Depression Scale (SDS). Measurements and Results HRV was significantly different among T1, T2, and T3. At T2, high frequency (HF) (normalized units of HF [nuHF]) was decreased and low frequency (LF) (normalized units of LF) and LF/HF were increased compared with those at T1 and T3. In the subgroup analysis between high and low SDS groups, high SDS group showed significantly decreased nuHF ( P = .035), increased nuLF ( P = .039), and increased LF/HF ( P = .020) compared to low SDS group at T1. However, these values at T2 and T3 were not different between 2 groups. In analysis within the groups, low SDS group showed significant differences in nuHF, nuLF, and LF/HF among T1, T2, and T3 ( P Conclusions HRV decreased significantly immediately before anesthesia and recovered to baseline with anesthetic induction. Preoperative, more depressed patients showed increased sympathetic tone at baseline and blunted response to impending anesthesia on the HRV measurements.

Technology is changing the way of navigation. New technologies for communication and navigation can be found on virtually every vessel. System architectures define structure and cooperation of components and subsystems. IMO, IALA, costal authorities, technology provider and many more actually propose new architectures for e-Navigation. This paper looks at other transportation domains and technical as normative requirements for e-Navigation architectures. With the aim of identifying possible synergies in the research, development, certification and standardization, this paper sets out to compare requirements and approaches of these two domains with respect to safety and security aspects. Since from an autonomy perspective, the automotive domain has started earlier and therefore has achieved a higher degree of technical progress, we will start with an overview of the developments in this domain. After that, the paper discusses the requirements on automation and assistance systems in the maritime domain and gives an overview of the developments into this direction within the maritime domain. This then allows us to compare developments in both domains and to derive recommendations for further developments in the maritime domain at the end of this paper.

Chinese hamster ovary (CHO) cell cultivation for production of therapeutic proteins is accompanied by production of metabolic wastes, mostly ammonia and lactate. To reduce ammonia production, the glutamine synthetase (GS) system was used to develop therapeutic monoclonal antibody (mAb)-producing CHO cells (SM-0.025). Additionally, the lactate dehydrogenase-A (LDH-A) was downregulated with shRNA to reduce lactate production in SM-0.025. The resulting mAb-producing cell lines (#2, #46, and #52) produced less ammonia than the host cell line during the exponential phase due to GS protein overexpression. LDH-A downregulation in SM-0.025 not only reduced lactate production but also further reduced ammonia production. Among the three LDH-A-downregulated clones, clone #2 had the highest mAb production along with significantly reduced specific lactate and ammonia production rates compared to those in SM-0.025. Waste reduction increased the galactosylation level of N-glycosylation, which improved mAb quality. LDH-A downregulation was also successfully applied to the host cell lines (CHO K1 and GS knockout CHO-K1). However, LDH-A downregulated host cells could not survive the pool-selection process wherein glutamine was excluded and methionine sulfoximine was added to the media. Taken together, LDH-A downregulation in the mAb-producing cell line generated with the GS system successfully reduced both ammonia and lactate levels, improving mAb galactosylation. However, LDH-A downregulation could not be applied to host cell lines because it hampered the selection process of the GS system.

The purpose of this study was to investigate whether and how topical nerve growth factor (NGF) attenuates streptozotocin (STZ)-induced diabetic cataracts in vivo. Rats were randomly divided into three groups, including the normal control rat group, STZ-induced diabetic cataract rat group (DM group), and STZ-induced diabetic cataract rat group treated with 200 μg/mL recombinant rat β-NGF (DM + NGF group). Cataract formation was evaluated by portable slit lamp biomicroscopy following pupil dilation at 8 weeks. The expression levels of NGF, aldose reductase (AR), and Na+/K+-ATPase in the lens epithelial cells (LECs) of the three groups were measured in the presence or absence of topical NGF. TUNEL-positive LECs were quantified to determine if hyperglycemia caused LEC apoptosis. At 8 weeks, the mean cataract score in the control group was significantly lower than that in DM and DM + NGF groups, and the score in the DM + NGF group was significantly lower than that in the DM group. At the equatorial zone and anterior central zone of lens, NGF and Na+/K+-ATPase expression levels were significantly decreased in the DM group; however, they were partially restored in the DM + NGF group. At the equatorial zone and anterior central zone of lens, AR expression and TUNEL-positive apoptotic LECs were significantly increased in the DM group compared with the control group, however, they were significantly decreased in the DM + NGF group. In conclusion, topical NGF could delay the progression of diabetic cataracts by attenuating polyol pathway activation and increasing Na+/K+-ATPase protein levels.

When designing a hand caliber with a high-temperature, high-pressure internal fluid transport pipe, reliability, safe use, and performance must be considered. Reliability refers to the stress caused by thermo-mechanical load, safe use refers to the low-temperature burns that might occur upon contact, and high-temperature burns caused by gas leakage occurring in the cylinder gap, and performance refers to projectile velocity. In this study, numerical simulation methods for heat transfer, structure analysis, and gas leakage are proposed so that solutions can be designed to account for the above three criteria. Furthermore, a hand-caliber design guide is presented. For heat transfer and structural analysis, mesh size, the transient convective heat transfer coefficient, and boundary conditions are described. Regarding gas leakage, methods reflecting projectile motion and determination of the molecular weight of the propellant are described. As a result, a designed hand caliber will have a high reliability, because the thermo-mechanical stress is lower than the yield stress. There will be little risk of low-temperature burns, but there will be a high temperature-burn risk, owing to gas leakage in the cylinder gap. The larger the cylinder-gap size, the greater the gas leakage and the smaller projectile velocity. The presented numerical simulation method can be applied to evaluate various aspects of other structures that require high-temperature, high-pressure fluid-transport pipes.

Generation of recombinant Chinese hamster ovary (rCHO) cell lines is critical for the production of therapeutic proteins. However, the high degree of phenotypic heterogeneity among generated clones, referred to as clonal variation, makes the rCHO cell line development process inefficient and unpredictable. Here, we investigated the major genomic causes of clonal variation. We found: (1) consistent with previous studies, a strong variation in rCHO clones in response to hypothermia (33 vs 37 oC) after random transgene integration; (2) altered DNA sequence of randomly integrated cassettes, which occurred during the integration process, affecting the transgene expression level in response to hypothermia; (3) contrary to random integration, targeted integration of the same expression cassette, without any DNA alteration, into three identified integration sites showed the similar response of transgene expression in response to hypothermia, irrespective of integration site; (4) switching the promoter from CMV to EF1α eliminated the hypothermia response; and (5) deleting the enhancer part of the CMV promoter altered the hypothermia response. Thus, we have revealed the effects of integration methods and cassette design on transgene expression levels, implying that rCHO cell line generation can be standardized through detailed genomic understanding. Further elucidation of such understanding is likely to have a broad impact on diverse fields that use transgene integration, from gene therapy to generation of production cell lines.

Purpose To evaluate the effect of diquafosol on corneal epithelium in a dry eye model using Transwell culture and a scopolamine-induced dry eye rat model. Methods Desiccation stress induced in an in vitro dry eye model using human corneal epithelial cells was used, and the cells were incubated with or without diquafosol media diluted at 1:100. Reactive oxygen species (ROS) generation was measured using 2',7'-dichlorofluorescein diacetate (DCFH-DA). Apoptosis was analyzed, and levels of phosphorylated Erk1/2, phosphorylated p90RSK, phosphorylated Akt, IκB-α, and NF-κB-p65 were determined. Levels of IL-1β, TNF-α, IL-6, IL-8, and GM-CSF were quantified. To investigate the in vivo effects of diquafosol, we induced dry eye in Wistar rats using scopolamine hydrobromide. The rats were divided into three groups: control, dry eye, and dry eye diquafosol; topical DIQUAS was applied four times daily for 28 days. We used immunohistochemistry to detect the levels of phosphorylated Erk1/2, phosphorylated p90RSK, and IL-1β, and used the TUNEL assay in corneal tissue. Results The distribution of highly fluorescent dichlorofluorescein and the proportion of annexin V- and PI-positive cells decreased in the diquafosol medium. Diquafosol increased the levels of phospho-Erk1/2, phospho-90RSK, phospho-Akt, and IκB-α, whereas it significantly decreased the levels of NF-κB-p65, IL-1β, and TNF-α. In vivo, apoptosis was enhanced in dry eye group. This response was markedly reduced and the level of phosphorylated p90RSK and phosphorylated ERK1/2 were upregulated and IL-1β was downregulated by DIQUAS. Conclusions Diquafosol treatment reduced intracellular ROS levels, apoptosis, and inflammation, all of which were increased in the dry eye model through desiccation.

Epidural blood patch (EBP) has been shown to be an effective treatment option for spontaneous intracranial hypotension (SIH). We investigated whether response to the EBP was related to the distance of the injection site from the leakage site in patients with SIH.We reviewed patients with SIH who underwent EBP at a single hospital. Patients were assigned to group R (response after EBP) or group N (no response after EBP). We then analyzed the demographics, clinical characteristics, leakage site, leakage length, EBP injection level and distance from leakage site, and injected EBP volume.Sixty-two patients were included in the analysis. The overall response rate to EBP was 59.7% (37 patients). The leakage length and injection distance from the leakage site did not differ between the two groups. Age, gender, body mass index, leakage site, and EBP volume did not differ significantly between the two groups.The clinical effect of EBP in SIH was not affected by leakage length or injection distance to leakage site. Further large studies must be conducted to investigate the efficacy of targeted EBP for SIH.

Chinese hamster ovary (CHO) cells are the preferred host for the production of a wide array of biopharmaceuticals. Thus, efficient and rational CHO cell line engineering methods have been in high demand to improve quality and productivity. Here, we provide a novel genome engineering platform for increasing desirable phenotypes of CHO cells based upon the integrative protocol of high-throughput RNA sequencing and DNA-free RNA-guided Cas9 (CRISPR associated protein9) nuclease-based genome editing. For commercial production of therapeutic proteins, CHO cells have been adapted for suspension culture in serum-free media, which is highly beneficial with respect to productivity and economics. To engineer CHO cells for rapid adaptation to a suspension culture, we exploited strand-specific RNA-seq to identify genes differentially expressed according to their adaptation trajectory in serum-free media. More than 180 million sequencing reads were generated and mapped to the currently available 109,152 scaffolds of the CHO-K1 genome. We identified significantly downregulated genes according to the adaptation trajectory and then verified their effects using the genome editing method. Growth-based screening and targeted amplicon sequencing revealed that the functional deletions of Igfbp4 and AqpI gene accelerate suspension adaptation of CHO-K1 cells. The availability of this strand-specific transcriptome sequencing and DNA-free RNA-guided Cas9 nuclease mediated genome editing facilitates the rational design of the CHO cell genome for efficient production of high quality therapeutic proteins.

PURPOSE The aim of this study was to investigate the effect of intracameral injection of bisulfite-containing phenylephrine on rabbit corneal endothelium cells in rabbits. METHODS Fifteen eyes of 15 New Zealand white rabbits were divided into 3 groups. Five eyes from each group were injected with 0.5% (0.5 mL, 0.05% bisulfite, group 1), 1.0% bisulfite-containing phenylephrine (0.5 mL, 0.1% bisulfite, group 2), or an equal volume of balanced salt solution as a control (group 3). Specular microscopy, corneal pachymetry, and a slit-lamp examination were used to evaluate corneal endothelial cell loss and other complications before injection, and at 1 hour and 1 week after injection. For morphological studies, corneal buttons were excised at 1 week and stained with alizarin red and trypan blue. Optical microscopy was used to examine 1 cornea from each of the 3 groups. RESULTS Within each group, there were no changes in specular microscopy and corneal pachymetry findings over the 1-week duration of the study. Although all groups showed the presence of several giant cells in the endothelium, the hexagonal shape of the corneal endothelium was morphologically preserved, and no abnormal endothelial cells were observed. CONCLUSIONS Intracameral injection of bisulfite-containing phenylephrine (up to 1.0%) does not affect endothelial cell viability or morphology of the rabbit cornea.