Mesenchymal Stem Cell-Extracellular Vesicle Therapy for Stroke: Scalable Production and Imaging Biomarker Studies

A major clinical hurdle to translate MSC-derived extracellular vesicles (EVs) is the lack of a method to scale-up the production of EVs with customized therapeutic properties. In this study, we tested whether EV production by a scalable 3D-bioprocessing method is feasible and improves neuroplasticity in animal models of stroke using MRI study. MSCs were cultured in a 3D-spheroid using a micro-patterned well. The EVs were isolated with filter and tangential flow filtration and characterized using electron microscopy, nanoparticle tracking analysis, and small RNA sequencing. Compared to conventional 2D culture, the production-reproduction of EVs (the number/size of particles and EV purity) obtained from 3D platform were more consistent among different lots from the same donor and among different donors. Several microRNAs with molecular functions associated with neurogenesis were upregulated in EVs obtained from 3D platform. EVs induced both neurogenesis and neuritogenesis via microRNAs (especially, miR-27a-3p and miR-132-3p)-mediated actions. EV therapy improved functional recovery on behavioral tests and reduced infarct volume on MRI in stroke models. The dose of MSC-EVs of 1/30 cell dose had similar therapeutic effects. In addition, the EV group had better anatomical and functional connectivity on diffusion tensor imaging and resting-state functional MRI in a mouse stroke model. This study shows that clinical-scale MSC-EV therapeutics are feasible, cost-effective, and improve functional recovery following experimental stroke, with a likely contribution from enhanced neurogenesis and neuroplasticity

Effects of anxiety, depression, social support, and physical health status on the health-related quality of life of pregnant women in post-pandemic Korea: a cross-sectional study

Purpose This study aimed to examine the effects of anxiety, depression, social support, and physical health status on the health-related quality of life of Korean pregnant women using Spilker’s quality of life model. Methods This was a cross-sectional study with a correlational design. The participants included 166 pregnant women who were recruited via convenience sampling at two healthcare centers in South Korea. Questionnaires were collected from April 22 to May 29, 2023, in two cities in South Korea. The EuroQol-5D-3L, General Anxiety Disorder-7, Patient Health Questionnaire-2, Perceived Social Support through Others Scale-8, and EuroQol visual analog scale were used to assess the study variables. The t-test, Pearson correlation coefficients, and multiple regression tests were conducted using IBM SPSS ver. 26.0. Results Statistically significant correlations were identified between the health-related quality of life of pregnant women and anxiety (r=.29, p<.001), depression (r=.31, p<.001), social support (r=–.34, p<.001), and physical health status (r=–.44, p<.001). Physical health status (β=–.31, p<.001) and social support (β=–.21, p=.003) had the greatest effect on health-related quality of life (F=15.50, p<.001), with an explanatory power of 26.0%. Conclusion The health-related quality of life of pregnant women was affected by social support and physical health status. This study demonstrated that physical health and social support promotion can improve the health-related quality of life of pregnant women. Healthcare providers should consider integrating physical health into social support interventions for pregnant women in the post-pandemic era

Real-Time Temporal Dynamics of Bicistronic Expression Mediated by Internal Ribosome Entry Site and 2A Cleaving Sequence

Multicistronic elements, such as the internal ribosome entry site (IRES) and 2A-like cleavage sequence, serve crucial roles in the eukaryotic ectopic expression of exogenous genes. For utilization of multicistronic elements, the cleavage efficiency and order of elements in multicistronic vectors have been investigated; however, the dynamics of multicistronic element-mediated expression remains unclear. Here, we investigated the dynamics of encephalomyocarditis virus (EMCV) IRES- and porcine teschovirus-1 2A (p2A)-mediated expression. By utilizing real-time fluorescent imaging at a minute-level resolution, we monitored the expression of fluorescent reporters bridged by either EMCV IRES or p2A in two independent cultured cell lines, HEK293 and Neuro2a. We observed significant correlations for the two fluorescent reporters in both multicistronic elements, with a higher correlation coefficient for p2A in HEK293 but similar coefficients for IRES-mediated expression and p2A-mediated expression in Neuro2a. We further analyzed the causal relationship of multicistronic elements by convergent cross mapping (CCM). CCM revealed that in all four conditions examined, the expression of the preceding gene causally affected the dynamics of the subsequent gene. As with the cross correlation, the predictive skill of p2A was higher than that of IRES in HEK293, while the predictive skills of the two multicistronic elements were indistinguishable in Neuro2a. To summarize, we report a significant temporal correlation in both EMCV IRES- and p2A-mediated expression based on the simple bicistronic vector and real-time fluorescent monitoring. The current system also provides a valuable platform to examine the dynamic aspects of expression mediated by diverse multicistronic elements under various physiological conditions.National Research Foundation of Korea (NRF) of the Ministry of Science and ICT [NRF-2017R1C1B2008775, NRF-2017R1A4A1015534, NRF-2018M3C7A1022310]; KBRI basic research program through Korea Brain Research Institute - Ministry of Science and ICT [17-BR-04]Open access journalThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Novel LMNA Gene Mutation in a Patient With Atypical Werner's Syndrome

Hutchinson-Gilford progeria syndrome (HGPS) and Werner's syndrome are representative types of progeroid syndrome. LMNA (Lamin A/C) gene mutation with atypical Werner's syndrome have recently been reported. Atypical Werner's syndrome with the severe metabolic complications, the extent of the lipodystrophy is associated with A133L mutation in the LMNA gene and these patients present with phenotypically heterogeneous disorders. We experienced a 15-yr-old Korean female with progeroid features, generalized lipodystrophy, hypertriglyceridemia, fatty liver, steatohepatitis, and type 2 diabetes mellitus. Skin fibroblasts from the patient showed marked abnormal nuclear morphology, compared with that from normal persons. Gene analysis revealed that this patient had T506del of exon 2 in the LMNA gene. We report here the first case of atypical Werner's syndrome with frameshift mutation that was caused by T506del

A lab-on-a-disc platform enables serial monitoring of individual CTCs associated with tumor progression during EGFR-targeted therapy for patients with NSCLC

Rationale: Unlike traditional biopsy, liquid biopsy, which is a largely non-invasive diagnostic and monitoring tool, can be performed more frequently to better track tumors and mutations over time and to validate the efficiency of a cancer treatment. Circulating tumor cells (CTCs) are considered promising liquid biopsy biomarkers; however, their use in clinical settings is limited by high costs and a low throughput of standard platforms for CTC enumeration and analysis. In this study, we used a label-free, high-throughput method for CTC isolation directly from whole blood of patients using a standalone, clinical setting-friendly platform. Methods: A CTC-based liquid biopsy approach was used to examine the efficacy of therapy and emergent drug resistance via longitudinal monitoring of CTC counts, DNA mutations, and single-cell-level gene expression in a prospective cohort of 40 patients with epidermal growth factor receptor (EGFR)-mutant non-small cell lung cancer. Results: The change ratio of the CTC counts was associated with tumor response, detected by CT scan, while the baseline CTC counts did not show association with progression-free survival or overall survival. We achieved a 100% concordance rate for the detection of EGFR mutation, including emergence of T790M, between tumor tissue and CTCs. More importantly, our data revealed the importance of the analysis of the epithelial/mesenchymal signature of individual pretreatment CTCs to predict drug responsiveness in patients. Conclusion: The fluid-assisted separation technology disc platform enables serial monitoring of CTC counts, DNA mutations, as well as unbiased molecular characterization of individual CTCs associated with tumor progression during targeted therapy

A Study on ESD Protection Circuit Applying Silicon Controlled Rectifier-Based Stack Technology with High Holding Voltage

In this study, an improved Electrostatic Discharge (ESD) protection circuit with low trigger voltage and high holding voltage is proposed. ESD has become a serious problem in the semiconductor process because the semiconductor density has become very high these days. Therefore, much research has been done to prevent ESD. The proposed circuit is a stacked structure of the new unit structure combined by the Zener Triggering (SCR ZTSCR) and the High Holding Voltage SCR (HHVSCR). The simulation results show that the proposed circuit has low trigger voltage and high holding voltage. And the stack technology is applied to adjust the various operating voltage. As the results, the holding voltage is 7.7 V for 2-stack and 10.7 V for 3-stack

A Concept Analysis of Cultural Nursing Competence

PURPOSE: The aim of this study was to conduct a concept analysis of cultural nursing competence. METHODS: Cultural nursing competence was analyzed using Rodgers&apos; evolutionary concept development method. A literature search using the keywords &quot;cultural nursing competence&quot;, &quot;intercultural nursing competence&quot;, &quot;cultural nursing&quot;, &quot;cultural health nursing&quot;, and &quot;cultural competence&quot; was conducted in PubMed, CINAHL, ERIC, and RISS on material published before 2015. Database and bibliographic searches yielded 35 records. RESULTS: Cultural nursing competence comprised cognitive, affective, and behavioral domains. The critical attributes of the concept were sensitivity, equality, and activity. The analysis identified the following dimensions: awareness, openness, and coherence. The consequences of cultural nursing competence were personal satisfaction and social justice. The definition contained competence on both an individual and social level. CONCLUSION: Cultural competency enhances quality of care by narrowing health disparities and increasing client satisfaction. The concept analysis of cultural nursing competence may offer an acceptable framework which can be used to develop psychometric tools of this concept and provide guidelines in nursing practice

Intracellular Calcium Spikes in Rat Suprachiasmatic Nucleus Neurons Induced by BAPTA-Based Calcium Dyes

Background: Circadian rhythms in spontaneous action potential (AP) firing frequencies and in cytosolic free calcium concentrations have been reported for mammalian circadian pacemaker neurons located within the hypothalamic suprachiasmatic nucleus (SCN). Also reported is the existence of "Ca2+ spikes" (i.e., [Ca2+]c transients having a bandwidth of 10～100 seconds) in SCN neurons, but it is unclear if these SCN Ca2+ spikes are related to the slow circadian rhythms. Methodology/Principal Findings: We addressed this issue based on a Ca2+ indicator dye (fluo-4) and a protein Ca2+ sensor (yellow cameleon). Using fluo-4 AM dye, we found spontaneous Ca2+ spikes in 18% of rat SCN cells in acute brain slices, but the Ca2+ spiking frequencies showed no day/night variation. We repeated the same experiments with rat (and mouse) SCN slice cultures that expressed yellow cameleon genes for a number of different circadian phases and, surprisingly, spontaneous Ca2+ spike was barely observed (<3%). When fluo-4 AM or BAPTA-AM was loaded in addition to the cameleon-expressing SCN cultures, however, the number of cells exhibiting Ca2+ spikes was increased to 13～14%. Conclusions/Significance: Despite our extensive set of experiments, no evidence of a circadian rhythm was found in the spontaneous Ca2+ spiking activity of SCN. Furthermore, our study strongly suggests that the spontaneous Ca2+ spiking activity is caused by the Ca2+ chelating effect of the BAPTA-based fluo-4 dye. Therefore, this induced activity seems irrelevant to the intrinsic circadian rhythm of [Ca2+]c in SCN neurons. The problems with BAPTA based dyes are widely known and our study provides a clear case for concern, in particular, for SCN Ca2+ spikes. On the other hand, our study neither invalidates the use of these dyes as a whole, nor undermines the potential role of SCN Ca2+ spikes in the function of SCN