Reprogramming anchorage dependency to develop cell lines for recombinant protein expression.

As the biopharmaceutical industry continues to mature in its cost-effectiveness and productivity, many companies have begun employing larger-scale biomanufacturing and bioprocessing protocols. While many of these protocols require cells with anchorage-independent growth, it remains challenging to induce the necessary suspension adaptations in many different cell types. In addition, although transfection efficiency is an important consideration for all cells, especially for therapeutic protein production, cells in suspension are generally more difficult to transfect than adherent cells. Thus, much of the biomanufacturing industry is focused on the development of new human cell lines with properties that can support more efficient biopharmaceutical production. With this in mind, we identified a set of "Adherent-to-Suspension Transition" (AST) factors, IKZF1, BTG2 and KLF1, the expression of which induces adherent cells to acquire anchorage-independent growth. Working from the HEK293A cell line, we established 293-AST cells and 293-AST-TetR cells for inducible and reversible reprogramming of anchorage dependency. Surprisingly, we found that the AST-TetR system induces the necessary suspension adaptations with an accompanying increase in transfection efficiency and protein expression rate. Our AST-TetR system therefore represents a novel technological platform for the development of cell lines used for generating therapeutic proteins.

Disrupted Rotational Perception During Simultaneous Stimulation of Rotation and Inertia.

Two vestibular signals, rotational and inertial cues, converge for the perception of complex motion. However, how vestibular perception is built on neuronal behaviors and decision-making processes, especially during the simultaneous presentation of rotational and inertial cues, has yet to be elucidated in humans. In this study, we analyzed the perceptual responses of 20 participants after pairwise rotational experiments, comprised of four control and four test sessions. In both control and test sessions, participants underwent clockwise and counterclockwise rotations in head-down and head-up positions. The difference between the control and test sessions was the head re-orientation relative to gravity after rotations, thereby providing only rotational cues in the control sessions and both rotational and inertial cues in the test sessions. The accuracy of perceptual responses was calculated by comparing the direction of rotational and inertial cues acquired from participants with that predicted by the velocity-storage model. The results showed that the accuracy of rotational perception ranged from 80 to 95% in the four control sessions but significantly decreased to 35 to 75% in the four test sessions. The accuracy of inertial perception in the test sessions ranged from 50 to 70%. The accuracy of rotational perception improved with repetitive exposure to the simultaneous presentation of both rotational and inertial cues, while the accuracy of inertial perception remained steady. The results suggested a significant interaction between rotational and inertial perception and implied that vestibular perception acquired in patients with vestibular disorders are potentially inaccurate.

HSPA5 and FGFR1 genes in the mesenchymal subtype of glioblastoma can improve a treatment efficacy.

Tyrosine kinase inhibitors (TKIs) have emerged as a potential treatment strategy for glioblastoma multiforme (GBM). However, their efficacy is limited by various drug resistance mechanisms. To devise more effective treatments for GBM, genetic characteristics must be considered in addition to pre-existing treatments. We performed an integrative analysis with heterogeneous GBM datasets of genomic, transcriptomic, and proteomic data from DepMap, TCGA and CPTAC. We found that poor prognosis was induced by co-upregulation of heat shock protein family A member 5 (HSPA5) and fibroblast growth factor receptor 1 (FGFR1). Co-up regulation of these two genes could regulate the PI3K/AKT pathway. GBM cell lines with co-upregulation of these two genes showed higher drug sensitivity to PI3K inhibitors. In the mesenchymal subtype, the co-upregulation of FGFR1 and HSPA5 resulted in the most malignant subtype of GBM. Furthermore, we found this newly discovered subtype was correlated with homologous recombination deficiency (HRD) In conclusion, we discovered novel druggable candidates within the group exhibiting co-upregulation of these two genes in GBM, suggest potential strategies for combination therapy.

Efficacy of vestibular rehabilitation and its facilitating and hindering factors from real-world clinical data.

Background and purpose: Customized vestibular rehabilitation improved dizziness and imbalance in several randomized controlled trials. In the present study, we determined the efficacy of customized vestibular rehabilitation using real-world observational data. Methods: In this retrospective observational study, we recruited 64 patients (median age = 60, interquartile range = 48-66.3) who completed the customized vestibular rehabilitation from January to December 2022. The outcomes of rehabilitation were evaluated using the dizziness handicap inventory (DHI) or vestibular disorders activities of daily living scale (VADL). The factors associated with outcomes were assessed with a generalized linear model, of which covariates included patients' age, sex, duration of illness, type of vestibular disorders, initial DHI and VADL scores, exercise compliance, and initial hospital anxiety and depression scale (HADS) scores. Results: After the median of 6 (4-6) weeks of rehabilitation, DHI and VADL scores significantly improved in patients with either peripheral or central vestibular disorders (Wilcoxon signed-rank test, p < 0.05). The initial DHI and VADL scores showed a positive while the sum of HADS scores showed a negative correlation with the outcome. In contrast, the age, sex, duration of illness, types of vestibular disorders, and exercise compliance did not affect the outcome. Discussion and conclusion: Customized vestibular rehabilitation is effective for central as well as peripheral disorders, especially when the symptoms are severe and the psychological distress is mild.

Integration of in vivo electrophysiology and optogenetics in rodents with PEDOT:PSS neural electrode array.

Here, we present a protocol for the fabrication of transparent implantable electrode arrays for integrating optogenetics and electrophysiology. We describe steps for fabricating microelectrodes using the conductive polymer poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate). We then detail procedures for analyzing performance of the electrodes and recording light-evoked neural activities from the transgenic mouse. This protocol utilizes photolithography rather than conventional electrodeposition. For complete details on the use and execution of this protocol, please refer to Cho et al. (2022).1.

Sustainable production of natural products using synthetic biology: Ginsenosides.

Synthetic biology approaches offer potential for large-scale and sustainable production of natural products with bioactive potency, including ginsenosides, providing a means to produce novel compounds with enhanced therapeutic properties. Ginseng, known for its non-toxic and potent qualities in traditional medicine, has been used for various medical needs. Ginseng has shown promise for its antioxidant and neuroprotective properties, and it has been used as a potential agent to boost immunity against various infections when used together with other drugs and vaccines. Given the increasing demand for ginsenosides and the challenges associated with traditional extraction methods, synthetic biology holds promise in the development of therapeutics. In this review, we discuss recent developments in microorganism producer engineering and ginsenoside production in microorganisms using synthetic biology approaches.

A Comparison Between GalT-/-;hCD39;hCD55 and GalT-/-;hCD39;hCD46;hCD55;TBM Pig Kidneys Transplanted in Nonhuman Primates.

Because there is a shortage of donor kidneys, researchers are exploring the possibility of using genetically modified pig kidneys for transplantation. Approaches involving knockout of carbohydrate genes or knockin of protective proteins have been attempted to determine the best gene modifications. In this study, we utilized GalT-/-;hCD39;hCD55 and GalT-/-;hCD39;hCD46;hCD55;thrombomodulin (TBM) pigs for transplantation in nonhuman primates (NHPs). The NHPs survived for 4 weeks after kidney transplantation (4 WAT) from the GalT-/-;hCD39;hCD55 pig and for 6 WAT from the GalT-/-;hCD39;hCD46;hCD55;TBM pig. However, messenger RNA (mRNA) sequencing and immunohistochemistry analysis revealed that the 6 WAT kidney exhibited more severe apoptosis, inflammation, loss of renal function, and renal fibrosis than the 4 WAT kidney. These results indicate that additional knockin of complement regulator (hCD46) and coagulation regulator (TBM) is not enough to prevent renal damage, suggesting that improved immune suppression is needed for more prolonged survival.

Clinical impact of metformin exposure during Staphylococcus aureus bacteremia in patients with diabetes mellitus.

PURPOSE: Increasing evidence has suggested that metformin may play positive roles in a wide range of infectious diseases. This study aimed to investigate the clinical impact of metformin exposure during Staphylococcus aureus bacteremia (SAB) in patients with diabetes. METHODS: A 3-year observational cohort study of 452 patients (aged ≥ 16 years) with SAB was performed at a tertiary care hospital. Metformin exposure was defined as receiving metformin during SAB, regardless of metformin use before the onset of bacteremia. RESULTS: Of 452 patients, 51 (11.3%) were classified in Group A (diabetes with metformin exposure), 115 (25.4%) in Group B (diabetes without metformin exposure), and 286 (63.3%) in Group C (no diabetes). The 30-day mortality rate in Group A was significantly lower than that in Group B (3.9% [2/51] versus 14.8% [17/115]; p = 0.04) and lower than that in Group C (3.9% [2/51] versus 17.1% [49/286]; p = 0.02). The mortality rates did not differ between Group B and Group C (14.8% [17/115] versus 17.1% [49/286]; p = 0.57). The rates of persistent and recurrent bacteremia were comparable among the three groups. Multivariate analysis indicated that metformin exposure was significantly associated with reduced mortality (adjusted odds ratio, 0.20; 95% confidence interval, 0.04-0.88; p = 0.03). CONCLUSIONS: Metformin exposure during SAB appears to be an independent predictor of survival in patients with diabetes.

Pelargonium sidoides extract mediates nephrotoxicity through mitochondrial malfunction and cytoskeleton destabilization.

We investigated the cytotoxic effect of Pelargonium sidoides extract on Madin-Darby canine kidney (MDCK) cells. P. sidoides extract decreased the cell viability in a dose dependent manner (> 0.2%). The extract of P. sidoides decreased the mitochondrial action potential, increased the number of reactive oxygen species (ROS) inside the cell, and caused nicotinamide adenine dinucleotide hydride (NADH) to be released, all of which are signs of mitochondrial dysfunction. The results of unbiased mRNA sequencing showed that 0.3% P. sidoides extract upregulates the apoptosis-related gene (BBC3). This finding was supported by immunoblot analysis of apoptosis signal pathways, which included Bcl-2, Bax, cytochrome C (CytC), cleaved caspase 3 (CC3), cleaved caspase 7 (CC7), cleaved caspase 9 (CC9) and cleaved PARP (CP). It is interesting to note that the elevated levels of Bax, CytC, CC3, CC7, and CC9, as well as CP, were suppressed by N-acetyl-L-cysteine (NAC) pretreatment, which points to ROS-mediated apoptosis. The small GTPases, RhoA, and Rac1/cdc42-GTP-bound active form were all lowered when P. sidoides extract was used. Also, RhoA-related cytoskeleton signals (ROCK, p-LIMK1/2, p-cofilin) and Rac1/cdc42-related signals (N-WASP, WAVE-2) were inhibited by P. sidoides extract. NAC or RhoA/Rac1/cdc42 activator pretreatment reduced P. sidoides extract-induced actin destabilization. In this work, P. sidoides extract promotes apoptosis by causing mitochondrial dysfunction and cytoskeleton disassembly.

Impact of post-treatment symptoms on supportive care needs among breast cancer survivors in South Korea.

Objective: The aim of this study is to investigate the factors influencing the supportive care needs of survivors of breast cancer who have completed major treatment. Methods: A total of 121 survivors of breast cancer from an online community in South Korea participated in this study. The study variables were supportive care needs, physical symptoms, anxiety, and depression. Independent t-tests, one-way Analysis of Variance (ANOVA), Pearson's correlation, and hierarchical regression analyses were performed. Results: The highest rankings of supportive care needs of survivors of breast cancer were medical system and information needs, patient care and support needs, psychological needs, sexual needs, and physical and daily life needs. Hierarchical regression analysis revealed that the participants' supportive care needs were explained by physical symptoms (P â€‹< â€‹0.001) and anxiety (P â€‹< â€‹0.001), accounting for 52.1% of the variance. Conclusions: Supportive care needs of survivors of breast cancer have a high level of medical system and information needs, and posttreatment conditions are related to high physical symptoms and anxiety. In the future, it will be necessary to identify supportive care needs and apply interventions to reduce their physical symptoms and anxiety.

Distribution of corpora amylacea in the human midbrain: using synchrotron radiation phase-contrast microtomography, high-field magnetic resonance imaging, and histology.

Corpora amylacea (CA) are polyglucosan aggregated granules that accumulate in the human body throughout aging. In the cerebrum, CA have been found in proximity to ventricular walls, pial surfaces, and blood vessels. However, studies showing their three-dimensional spatial distribution are sparse. In this study, volumetric images of four human brain stems were obtained with MRI and phase-contrast X-ray microtomography, followed up by Periodic acid Schiff stain for validation. CA appeared as hyperintense spheroid structures with diameters up to 30 µm. An automatic pipeline was developed to segment the CA, and the spatial distribution of over 200,000 individual corpora amylacea could be investigated. A threefold-or higher-density of CA was detected in the dorsomedial column of the periaqueductal gray (860-4,200 CA count/mm3) than in the superior colliculus (150-340 CA count/mm3). We estimated that about 2% of the CA were located in the immediate vicinity of the vessels or in the peri-vascular space. While CA in the ependymal lining of the cerebral aqueduct was rare, the sub-pial tissue of the anterior and posterior midbrain contained several CA. In the sample with the highest CA density, quantitative maps obtained with MRI revealed high R2* values and a diamagnetic shift in a region which spatially coincided with the CA dense region.

The Significance of Digital Citizenship and Gender in the Relationship between Social Media Usage Time and Self-Esteem among Adolescents: A Secondary Analysis.

This study examined the significance of digital citizenship and gender in the relationship between social media usage time and self-esteem among adolescents. This cross-sectional study was a secondary analysis using national data on 506 Korean adolescents acquired from the 2020 Korea Media Panel Survey. The data were analyzed using SPSS 23.0 and SPSS PROCESS macro. We found that the moderated moderating effects of digital citizenship and gender on the relationship between social media usage time and self-esteem were significant. Both boys and girls had higher self-esteem in groups with higher digital citizenship than in groups with lower digital citizenship. The relationship between social media usage time and self-esteem for boys was positive in the high digital citizenship group and negative in the low digital citizenship group. Conversely, for girls, the relationship between social media usage time and self-esteem was positive in the low digital citizenship group and negative in the high digital citizenship group. It is important to take a differentiated approach that considers the relationship between digital citizenship and gender to promote healthy digital media use and positive self-esteem.

An Integrative Review of the Influence on Human Papillomavirus Vaccination Adherence among Adolescents.

To enhance the vaccination rate of adolescents against HPV, understanding their current vaccination intentions and identifying the factors that influence their decision to vaccinate are imperative. The Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) was used to search for relative literature. Five electronic databases (CINAHL, PubMed, Web of science, Research Information Sharing Service (RISS) and Korea Citation Index (KCI)) were searched from 2007 to 2022, and 19 studies were selected for final review. The Mixed Methods Assessment Tool (MMAT) was used as a critical appraisal tool to evaluate the quality for various types of research designs. This integrative review revealed three themes, including facilitators (personal characteristics and environmental factors), barriers (lack of awareness and knowledge about HPV, concerns about side effects, and the burden of costs), and ways to increase vaccination uptake (knowledge promotion and HPV vaccines coverage by public insurance). To encourage HPV vaccine uptake, it is essential to enhance public knowledge, create compelling advertisements, and ensure that the vaccines are covered by public insurance. Moreover, providing educational programs that emphasize the effectiveness and significance of HPV vaccines to adults who can influence the vaccination decisions of adolescents is vital.

A wireless, solar-powered, optoelectronic system for spatial restriction-free long-term optogenetic neuromodulations.

Numerous wireless optogenetic systems have been reported for practical tether-free optogenetics in freely moving animals. However, most devices rely on battery-powered or coil-powered systems requiring periodic battery replacement or bulky, high-cost charging equipment with delicate antenna design. This leads to spatiotemporal constraints, such as limited experimental duration due to battery life or animals' restricted movement within specific areas to maintain wireless power transmission. In this study, we present a wireless, solar-powered, flexible optoelectronic device for neuromodulation of the complete freely behaving subject. This device provides chronic operation without battery replacement or other external settings including impedance matching technique and radio frequency generators. Our device uses high-efficiency, thin InGaP/GaAs tandem flexible photovoltaics to harvest energy from various light sources, which powers Bluetooth system to facilitate long-term, on-demand use. Observation of sustained locomotion behaviors for a month in mice via secondary motor cortex area stimulation demonstrates the notable capabilities of our device, highlighting its potential for space-free neuromodulating applications.

Development of novel recombinant peroxidase secretion system from Pseudomonas putida for lignin valorisation.

Pseudomonas putida is a promising strain for lignin valorisation. However, there is a dearth of stable and efficient systems for secreting enzymes to enhance the process. Therefore, a novel secretion system for recombinant lignin-depolymerising peroxidase was developed. By adopting a flagellar type III secretion system, P. putida KT-M2, a secretory host strain, was constructed and an optimal secretion signal fusion partner was identified. Application of the dye-decolourising peroxidase of P. putida to this system resulted in efficient oxidation activity of the cell-free supernatant against various chemicals, including lignin model compounds. This peroxidase-secreting strain was examined to confirm its lignin utilisation capability, resulting in the efficient assimilation of various lignin substrates with 2.6-fold higher growth than that of the wild-type strain after 72 h of cultivation. Finally, this novel system will lead efficient bacterial lignin breakdown and utilization through enzyme secretion, paving the way for sustainable lignin-consolidated bioprocessing.

The influence of dietary patterns on skin bacterial diversity, composition, and co-occurrence relationships at forearm and neck sites of healthy Korean adults.

AIMS: Diet and nutrition are important aspects of skin physiology and health. However, the influence of diet on the bacterial flora of different skin sites is not well understood. Therefore, we investigated the relationship between dietary patterns (DPs) and skin bacterial flora on the forearm (a dry site) and the neck (a sebaceous site) of healthy Korean adults. METHODS AND RESULTS: In metagenomics analysis, Shannon and Simpson indices were higher on the forearm than on the neck and were negatively correlated with the two dominant species, Cutibacterium acnes and Staphylococcus epidermidis, on two skin sites. In addition, the Simpson index of the forearm was positively associated with DP1 (characterized by a high intake of vegetables, mushrooms, meat, fish and shellfish, seaweed, and fat and oil), while that on the neck was negatively associated with DP2 (characterized by a high intake of fast food). A high intake of DP1 was associated with a lower abundance of dominant species, including C. acnes, and higher degrees of the co-occurrence network, whereas a high intake of DP2 was associated with the opposite pattern. CONCLUSIONS: Specific diets may impact both skin bacterial diversity and composition, as well as the co-occurrence of bacteria, which may vary across different skin sites.

Functional rehabilitation of the maxillary sinus after modified endoscopic sinus surgery for displaced dental implants.

PURPOSE: Dental implants may become displaced into the maxillary sinus due to insufficient primary stability, changes in nasal air pressure, or surrounding bone resorption and should be removed as soon as possible. The aim of this study was to evaluate the efficacy of the modified endoscopic sinus surgery (MESS) approach for removal of displaced dental implants. METHODS: From September 2010 to November 2021, we studied 15 cases with displaced implants in the maxillary sinus. The patient characteristics, medical history, clinical and imaging results, and post-removal outcomes were retrospectively assessed. RESULTS: The symptoms included sinusitis (100%), pain (26.6%), postnasal drip (6.6%), nasal obstruction (26.6%), and oroantral communication (26.6%). Two cases were managed through the crestal approach (13.3%), while two cases were treated with the Caldwell-Luc procedure (13.3%). One case was addressed using functional endoscopic sinus surgery (6.7%), while 10 cases were managed with the MESS approach (66.7%). MESS allows functional rehabilitation of mucociliary clearance by the cilia in the sinus membrane. Implant displacement into the maxillary sinus can be classified as early, late, or delayed displacement. CONCLUSIONS: MESS is a reliable treatment option that can identify migrated dental implants in any part of the sinus with endoscopic assistance for functional rehabilitation of the maxillary sinus without postoperative sequelae.

Digital Citizenship and Life Satisfaction in South Korean Adolescents: The Moderated Mediation Effect of Poverty.

This study examined the moderated mediation effect of poverty on the paths between enactive mastery experience in digital life and life satisfaction mediated by digital citizenship and digital life among Korean adolescents using structural equation modelling. This cross-sectional study involved a secondary data analysis of 2020 national data in The Report on the Digital Divide provided by the National Information Society Agency (NIA) of Korea. Data from 1084 Korean adolescents were analyzed using IBM SPSS Statistics for Windows, version 26.0 and SPSS PROCESS macro. The results demonstrated a significant moderated mediation effect of poverty. Enactive mastery experience, which encompasses the self-knowledge, perceived task difficulty, and contextual factors of adolescents living in poverty, was associated with digital life and life satisfaction through the mediation of digital citizenship. For adolescents living in poverty, in contrast to their non-poor counterparts, enactive mastery experience in digital life and digital citizenship are two critical factors in life satisfaction. Therefore, institutional support enabling adolescents and their communities to forge partnerships is necessary to foster these two factors.

[The Effect of Socioeconomic Status to Depression of Perimenopause Women: Pathway Analysis Using the Reserve Capacity Model].

PURPOSE: Based on the Reserve Capacity Model, this study investigated the effects of pre- and postmenopausal women's socioeconomic status (SES) on depression, focusing on the mediating effects of self-esteem, happiness, and family relationship satisfaction with social network relationships. METHODS: This cross-sectional study involved secondary analysis of national data on 771 perimenopause women gathered from the 16th Korea Welfare Panel Study (KOWEPS) 2021. A path analysis model was constructed to evaluate the relationship between SES, social network satisfaction, self-esteem, perceived health status, and depression. Data were analyzed using ADANCO 2.3.1 and Mplus 8.4. RESULTS: Although SES had no direct effect on depression, it did affect depression through self-esteem, happiness, and satisfaction with family relationships. CONCLUSION: The findings of this study indicate that perimenopausal women's personal resources-psychosocial variables such as self-esteem and happiness-had a higher effect on depression than tangible reserves like SES. Therefore, interventions for enhancing self-esteem and happiness may prevent depression in perimenopausal women effectively.