Distinct properties of putative trophoblast stem cells established from somatic cell nuclear-transferred pig blastocysts.

BACKGROUND: Genetically modified pigs are considered ideal models for studying human diseases and potential sources for xenotransplantation research. However, the somatic cell nuclear transfer (SCNT) technique utilized to generate these cloned pig models has low efficiency, and fetal development is limited due to placental abnormalities. RESULTS: In this study, we unprecedentedly established putative porcine trophoblast stem cells (TSCs) using SCNT and in vitro-fertilized (IVF) blastocysts through the activation of Wing-less/Integrated (Wnt) and epidermal growth factor (EGF) pathways, inhibition of transforming growth factor-ß (TGFß) and Rho-associated protein kinase (ROCK) pathways, and supplementation with ascorbic acid. We also compared the transcripts of putative TSCs originating from SCNT and IVF embryos and their differentiated lineages. A total of 19 porcine TSCs exhibiting typical characteristics were established from SCNT and IVF blastocysts (TSCsNT and TSCsIVF). Compared with the TSCsIVF, TSCsNT showed distinct expression patterns suggesting unique TSCsNT characteristics, including decreased mRNA expression of genes related to apposition, steroid hormone biosynthesis, angiopoiesis, and RNA stability. CONCLUSION: This study provides valuable information and a powerful model for studying the abnormal development and dysfunction of trophoblasts and placentas in cloned pigs.

Pre-treatment quality of life in patients with salivary gland cancer in comparison with those of head and neck cancer patients.

INTRODUCTION: Salivary gland cancer (SGC) is a rare malignant tumor arising from the salivary glands, with a variety of clinical and biological behaviors different from head and neck cancer (HNC). Because of the rarity of SGC, there are limited data on pre-treatment quality of life (QoL). Therefore, we evaluated the pre-treatment QoL in SGC patients by stage and compared it with that of HNC patients. METHODS: From a prospective registry of HNC patients (2016-2020), we selected 225 patients with SGC, and 912 patients with oral cavity (OC) and oropharyngeal cancer (OPC) who were diagnosed in the same period as the HNC control group. The European Organization for Research and Treatment of Cancer (EORTC) Quality of Life Questionnaire Core 30 (QLQ-C30) and HNC-Specific Module (H&N35) were used to assess QoL. RESULTS: SGC patients had a statistically better baseline QoL (69.8 vs. 64.0), emotional (82.1 vs. 78.8), cognitive (92.0 vs. 88.7), and social function (86.3 vs. 80.5), and fewer symptoms than HNC patients. The estimated average QoL differences between SGC patient diagnosed at stages I and IV was -12.9. Especially, advanced-stage of tumors was associated with much lower role functioning and emotional functioning scores in SGC patients, compared to those in HNC patients, among females and of younger age. DISCUSSION: Although the overall QoL score was higher in SGC patients than in HNC patients, specific domains were significantly affected in SGC patients according to the tumor stage. Females and those of younger age were more affected by severity of disease in SGC. STUDY REGISTRATION: ClinicalTrials.gov Identifier NCT02546895.

Long-Term Efficacy and Safety of Eculizumab in Patients With Paroxysmal Nocturnal Hemoglobinuria and High Disease Burden: Real-World Data From Korea.

BACKGROUND: Paroxysmal nocturnal hemoglobinuria (PNH) is a rare hematologic disorder characterized by uncontrolled terminal complement activation. Eculizumab, a monoclonal antibody C5 inhibitor was introduced in Korea in 2009 and has been the standard treatment option for PNH. METHODS: This study assessed the long-term efficacy/safety of eculizumab in PNH using real-world data from the Korean Health Insurance Review and Assessment Service. Eighty patients who initiated eculizumab from 2009-2020 were enrolled. RESULTS: At eculizumab initiation, the median age was 51.5 years, lactate dehydrogenase (LDH) 6.8 × upper limit of normal, and granulocyte clone size 93.0%. All patients had at least one PNH-related complication before eculizumab initiation, including renal failure (n = 36), smooth muscle spasm (n = 24), thromboembolism (n = 20), and pulmonary hypertension (n = 15). The median (range) duration of eculizumab treatment was 52.7 (1.0, 127.3) months (338.6 total treated patient-years). Despite high disease activity in the study population before treatment initiation, overall survival was 96.2% and LDH levels were stabilized in most patients during treatment. PNH-related complications at treatment initiation were resolved in 44.4% of patients with renal failure, 95.8% with smooth muscle spasm, 70.0% with thromboembolism, and 26.7% with pulmonary hypertension. Extravascular hemolysis occurred in 28.8% of patients (n = 23; 0.09 per patient-year) and breakthrough hemolysis in 18.8% (n = 15; 0.06 per patient-year). No treatment discontinuation cases related to eculizumab were observed. CONCLUSION: These data provided evidence for the long-term efficacy and safety of eculizumab in Korean PNH patients with high disease burdens.

Advancements in 2D and 3D In Vitro Models for Studying Neuromuscular Diseases.

Neuromuscular diseases (NMDs) are a genetically or clinically heterogeneous group of diseases that involve injury or dysfunction of neuromuscular tissue components, including peripheral motor neurons, skeletal muscles, and neuromuscular junctions. To study NMDs and develop potential therapies, remarkable progress has been made in generating in vitro neuromuscular models using engineering approaches to recapitulate the complex physical and biochemical microenvironments of 3D human neuromuscular tissues. In this review, we discuss recent studies focusing on the development of in vitro co-culture models of human motor neurons and skeletal muscles, with the pros and cons of each approach. Furthermore, we explain how neuromuscular in vitro models recapitulate certain aspects of specific NMDs, including amyotrophic lateral sclerosis and muscular dystrophy. Research on neuromuscular organoids (NMO) will continue to co-develop to better mimic tissues in vivo and will provide a better understanding of the development of the neuromuscular tissue, mechanisms of NMD action, and tools applicable to preclinical studies, including drug screening and toxicity tests.

Flavones: The Apoptosis in Prostate Cancer of Three Flavones Selected as Therapeutic Candidate Models.

Cancer is a widespread but dangerous disease that can strike anyone and is the second 1leading cause of death worldwide. Prostate cancer, in particular, is a prevalent cancer that occurs in men, and much research is being done on its treatment. Although chemical drugs are effective, they have various side effects, and accordingly, anticancer drugs using natural products are emerging. To date, many natural candidates have been discovered, and new drugs are being developed as drugs to treat prostate cancer. Representative candidate compounds that have been studied to be effective in prostate cancer include apigenin, acacetin and tangeretin of the flavone family among flavonoids. In this review, we look at the effects of these three flavones on prostate cancer cells via apoptosis in vitro and in vivo. Furthermore, in addition to the existing drugs, we suggest the three flavones and their effectiveness as natural anticancer agents, a treatment model for prostate cancer.

Radiation dose evaluation to organs in neonatal patients by field size during potable X-ray examination in incubators: A Monte Carlo simulation study.

BACKGROUND: Neonatal patients located in incubators are exposed to as many as 159 radiographs until discharge. To reduce the dose exposed to the patient, factors that may cause unnecessary exposure to the patient were judged. When conducting portable X-rays of neonatal patients located in an incubator, it is not easy to determine the exact field size because collimation light is exposed on the acrylic plate, an incubator canopy, and the resulting shadow is reflected on the patient's body. OBJECTIVE: This study aims to measure the organ dose exposed to the patient according to the field size when a portable radiograph is given to a neonatal patient in a neonatal intensive care unit (NICU) incubator. METHODS: To identify the absorbed organ dose depending on the radiation field size during portable X-ray examination of neonatal patient, a Monte Carlo N-Particle (MCNP) simulation, a SpeckCalc program, and a neonatal phantom from the ICRP 89 are applied for the calculation. According to the minimal field size (MinFS) standards of the European Commission (EC), the smaller field size is intended to measure tightly from the top of the lung apices to the bottom of the genitals; a larger field size is also calculated by adding 6 cm in width and length. RESULTS: Compared to the hospital C condition from the previous study, the larger and smaller field sizes are decreased by an average of 45% and 67%, respectively. Study results also show a 42% reduction in smaller field size compared to the larger field size. CONCLUSION: When taking chest and abdomen radiographic images of neonatal patients in incubators, appropriate field sizes are required to prevent inappropriate dose absorption for non-thoracic organs.

Exploring the mechanism of trehalose: dual functions of PI3K/Akt and VPS34/mTOR pathways in porcine oocytes and cumulus cells†.

Autophagy, an intracellular recycling system, is essential for the meiotic maturation of porcine oocytes. Trehalose has been reported as a novel mammalian target of rapamycin (mTOR)-independent autophagy inducer in many cells. Furthermore, we previously have demonstrated that trehalose supplementation during in vitro maturation of porcine oocytes improves the developmental competence of parthenogenetic embryos, possibly via autophagic activation, whereas the underlying mechanisms remain unclear. Therefore, the aim of this study was to address this issue. We found that trehalose plays a role as an autophagy activator by autophagic flux assay and determined that it promotes phosphatidylinositol-3 kinase (PI3K)/protein kinase B (Akt) inhibition and vacuolar protein sorting 34 (VPS34)/mTOR activation by immunoblotting, both in cumulus cells (CCs) and oocytes. However, interestingly, the effects and the mechanisms regulated by trehalose were different in them, respectively. In CCs, the autophagy was activated through the improvement of lysosomal function/autophagic clearance viability by upregulation of coordinated lysosomal expression and regulation genes via PI3K/Akt inhibition. Whereas in oocytes, autophagy was activated via induction of VPS34, which directly influences autophagosome formation, and the precise meiotic process was ensured via Akt inhibition and mTOR activation. Taken together, this study furtherly elucidates the novel detailed mechanism of trehalose during porcine oocyte maturation, thus laying the biological foundations for pharmacological application.

Amine-modified nanoplastics promote the procoagulant activation of isolated human red blood cells and thrombus formation in rats.

BACKGROUND: Microplastics (MPs) and nanoplastics (NPs) formed from decomposed plastic are increasing environmental threats. Although MPs and NPs exposed through various routes enter the systemic circulation, the potential toxicity of those is largely unknown. We investigated whether polystyrene NPs (PS-NPs) promote the coagulation activity of red blood cells (RBCs). RESULTS: We tested several types of PS-NPs using human RBCs and found that amine-modified 100 nm PS-NPs were the most potent. We measured the uptake of PS-NPs using flow cytometry and confocal microscopy. Electron microscopy revealed morphological changes of RBCs by PS-NPs. PS-NPs induced the externalization of phosphatidylserine, generation of microvesicles in RBCs, and perturbations in the intracellular microenvironment. PS-NPs increased the activity of scramblases responsible for phospholipid translocation in RBCs. PS-NPs modulated the functional interaction to adjacent tissues and coagulation cascade, enhancing RBC adhesion and thrombin generation. Our observations in human RBCs were consistent with those in isolated rat RBCs, showing no inter-species differences. In rat venous thrombosis models, the intravenous administration of PS-NPs enhanced thrombus formation. CONCLUSION: Amine-modified PS-NPs induce the prothrombotic activation of RBCs causing thrombus formation. We believe that our study will contribute to understanding the potential toxicity of amine-modified polystyrene particles in blood cells and cardiovascular systems.

Community leaders' perceptions of depression and the perceived barriers in seeking mental health services for older Korean Americans.

OBJECTIVES: This study explored community leaders' understanding of depression among older Korean Americans and barriers to seeking mental health services. Depression is prevalent among older Korean Americans, but they are less likely to seek help from professionals and prefer to use informal methods. Older Korean Americans strongly prefer to maintain their ethnic traditions and use community service agencies provided by their same ethnicity. In this regard, community leaders who provide services for older adults play a significant role in not only advocating for the population but also in developing services and programs for their communities. DESIGN: In this qualitative study, 12 Korean community leaders who provide services for older adults were interviewed. Data were analyzed thematically, and Nvivo 12 was used to organize the data and to detect relevant themes. RESULTS: Findings showed that community leaders had an in-depth understanding of the causes of depression from environmental and cultural contexts including isolation, losing independence, cultural factors, and lack of family support. However, although the leaders were aware of the severity of depression among older Korean Americans, they did not understand biologically-based factors; rather, they considered it a natural part of aging. Findings indicated that they were biased towards older adults like older Korean Americans, attributing depressive symptoms to personality issues. They also heavily rely on personal experiences to understand depression instead of on having educational training. Barriers to getting help for older adults include lack of the perceived need of older adults, lack of professionals, and a prevalent stigma against depression that encourages individuals to hide their illness. CONCLUSIONS: The findings indicated that community-based educational training is necessary to increase understanding of depression not only for the community leaders but also for individuals and families.

Community agency directors' attitudes on depression treatment among older Korean Americans and barriers to providing services.

Objectives: The purpose of this study is to explore Korean agency directors' attitudes about depression treatment and their current efforts and barriers to providing services for older Korean Americans.Methods: Interviews were conducted with 11 agency directors, providing services from ethnic community centers and health care agencies. A constant comparative approach was applied to detect emerging themes.Results: Findings revealed that the agency directors had a negative attitude about using antidepressants and counseling. Based on these beliefs, they were less likely to recommend individuals with depression to use medical treatment or counseling; instead, they recommended informal ways of treating depression, including reliance on peers, family, and religious activities. A lack of educational training, funding, collaboration, and Korean-speaking professionals were perceived to be barriers to providing services.Discussion: Findings indicate the importance of educational training to increase understanding of depression treatment and to support the agency directors in offering services for older Korean Americans.

Prevention of partner violence: virtual case simulation for religious leaders of Korean American immigrant communities.

The project 'Religious Leaders for Healthy Families' aims to prevent intimate partner violence (IPV) and increase access to resources for immigrant victims by enhancing the capacity of religious leaders. Due to cultural and psychosocial barriers, immigrant women prefer to exhaust informal resources, including religious leaders, before seeking help from professional service providers. This study reports the development of a virtual case simulation in which Korean American (KA) religious leaders living in the USA practice how to prevent and address IPV in their congregation. Intervention mapping (IM) guided the development of the simulation. This process was informed by theory, research and expertise in prevention and virtual case simulation. We partnered with victim service organizations and received feedback from religious leaders. The simulation was pilot tested with nine KA religious leaders and three community leaders with IPV expertise in the KA community. The resulting intervention consists of 4 behavioral outcomes and 24 performance objectives. We identified the knowledge, attitudes, outcome expectations and self-efficacy needed to accomplish each objective. The most creative phase was the development of four modules, as interactive virtual case simulations, that address all performance objectives. IM and social cognitive theory provided a useful framework for developing this virtual case simulation. With culturally responsive modifications, the intervention has the potential to be adapted for religious leaders from other immigrant communities.

Religion can play a pivotal role in helping immigrant women. Religious institutions help immigrants maintain their ethnic identity and language and support their integration into the new culture. This study explains the development of an online intervention for Korean American religious leaders to prevent partner violence. Religious leaders in immigrant communities can play a significant role in perpetuating cultural norms that contribute to partner violence or, conversely, create norms that promote equity between partners and value seeking help. Partner violence is high in cultures that adhere to traditional gender roles, with male-dominated interactions, and perceive seeking help as a failure that would bring shame to the whole family. The intervention consists of four modules that depict interactions between a religious leader and female parishioners: (i) a deacon worried about a parishioner who might be a victim, (ii) an immigrant woman married to an abusive husband, (iii) a woman in the hospital because of multiple bruises and a broken arm and (iv) a graduate student concerned about her fiancé's behavior. Experts in various fields, community leaders in the prevention of partner violence, and religious leaders provided their feedback and expertise for program development.

Monitoring clinical exposure index and deviation index for dose optimization based on national diagnostic reference level: Focusing on general radiography of extremities.

BACKGROUND: The International Electrotechnical Commission established the concept of the exposure index (EI), target exposure index (EIT) and deviation index (DI). Some studies have conducted to utilize the EI as a patient dose monitoring tool in the digital radiography (DR) system. OBJECTIVE: To establish the appropriate clinical EIT, this study aims to introduce the diagnostic reference level (DRL) for general radiography and confirm the usefulness of clinical EI and DI. METHODS: The relationship between entrance surface dose (ESD) and clinical EI is obtained by exposure under the national radiography conditions of Korea for 7 extremity examinations. The EI value when the ESD is the DRL is set as the clinical EIT, and the change of DI is then checked. RESULTS: The clinical EI has proportional relationship with ESD and is affected by the beam quality. When the clinical EIT is not adjusted according to the revision of DRLs, there is a difference of up to 2.03 in the DI value and may cause an evaluation error of up to 1.6 times for patient dose. CONCLUSIONS: If the clinical EIT is periodically managed according to the environment of medical institution, the appropriate patient dose and image exposure can be managed based on the clinical EI, EIT, and DI.

Evaluation of radiation dose to organs of neonatal patients during portable X-ray examination in incubators: A Monte Carlo simulation study.

BACKGROUND: Infants admitted to neonate intensive care units (NICUs) are placed in incubators to maintain body temperature and condition, which undergo normal radiographs and are exposed to radiation. Furthermore, different incubator structures in different hospitals exhibit varying object to image receptor distance (OID), source to image receptor distance (SID), presence of canopy, which results in variations in X-ray radiation conditions and doses absorbed by the neonatal patients. OBJECTIVE: To measure organ dose exposed to neonatal patient in different incubator settings. METHODS: A portable X-ray was performed on a neonatal patient placed in an incubator to identify disease progress, the injection path of the drug, and various factors. To minimize direct contact between neonatal patients and image receptor, radiologic technologists place the image receptor on a tray underneath the incubator and place the portable X-ray tube on top of the acrylic canopy of the incubators. SID and OID settings and value of organ dose exposed to the patient varied based on the incubator structure, and the organ absorbed dose was determined using Monte Carlo N-Particle (MCNP) simulation, PC-based Monte Carlo program (PCXMC) 2.0 simulation, and neonate phantoms. RESULTS: Evaluations of organ dose of neonatal patients in three hospitals with different incubator settings reveal that the average organ dose differs by 36% depending on change in OID and SID settings and reduces by 10% with an acrylic canopy. Therefore, owing to the presence of an acrylic canopy on the top of the incubator and the longer SID with the corresponding shorter OID, a lower dose was absorbed by organs of neonatal patient. CONCLUSION: Our results provide proof that proper incubator standard decreases organ dose to neonatal patient during continuously diagnostic X-ray procedure.

Clinical outcomes of bulky parotid gland cancers: need for self-examination and screening program for early diagnosis of parotid tumors.

BACKGROUND: Early detection and diagnosis of parotid gland cancer (PGC) are essential to improve clinical outcomes, because Tumor-Node-Metastasis stage at diagnosis is a very strong indicator of prognosis in PGC. Nevertheless, some patients still present with large parotid mass, maybe due to the unawareness or ignorance of their disease. In this study, we aimed to present the clinical outcomes of bulky PGC (defined by a 4 cm cutoff point for T3-4 versus T1-2 tumors), to emphasize the necessity of a self-examination tool for parotid gland tumor. METHODS: We retrospectively reviewed 60 consecutive cases with bulky (equal to and greater than 4 cm in the longest diameter, determined radiologically) malignant tumors arising from the parotid gland from 1995 to 2016. The clinical and pathological factors were analyzed to identify risk factors for poor outcomes using Cox proportional hazard models. In addition, we designed a self-examination tool for parotid gland tumors, similar to breast self-examination for breast cancer detection. RESULTS: Patients with bulky parotid cancer showed 48.9% 5-year and 24.5% 10-year overall survival rates and a 47.9% risk of high-grade malignancy. The common pathological diagnoses were carcinoma ex pleomorphic adenoma (18.3%), adenocarcinoma (16.7%), mucoepidermoid carcinoma (16.7%), salivary duct carcinoma (16.7%), and adenoid cystic carcinoma (11.7%). Survival analyses revealed that tumor size (hazard ratio, HR = 1.262 upon increase of 1 cm, 95% confidence interval, 95%CI 1.059-1.502), lymph node metastasis (HR = 2.999, 95%CI 1.048-8.583), and high tumor grade (HR = 4.148, 95%CI 1.215-14.154) were independent prognostic factors in multivariable analysis. Functional preservation of the facial nerve was possible only in less than half of patients. CONCLUSION: In bulky PGC, lymph node metastasis at diagnosis and high tumor grade indicated poor survival outcomes, and functional outcomes of the facial nerve were suboptimal. Thus, a public effort seems to be necessary to decrease these patients with bulky PGC, and to increase patients' self-awareness of their disease. As a way of early detection, we proposed a parotid self-examination tool to detect parotid gland tumors at an early stage, which is similar to breast self-examination.

Particulate Nitrosamines and Nitramines in Seoul and Their Major Sources: Primary Emission versus Secondary Formation.

Seven nitrosamines and three nitramines in particulate matter with an aerodynamic diameter of less than or equal to 2.5 µm (PM2.5) collected in 2018 in Seoul, South Korea, were quantified. Annual mean concentrations of the sum of nitrosamines and nitramines were 9.81 ± 18.51 and 1.12 ± 0.70 ng/m3, respectively, and nitrosodi-methylamine (NDMA) and dimethyl-nitramine (DMN) comprised the largest portion of nitrosamines and nitramines, respectively. Statistical analyses such as non-parametric correlation analysis, positive matrix factorization, analysis of covariance, and orthogonal partial least squared discrimination analysis were carried out to identify contribution of the atmospheric reactions in producing NDMA and DMN. In addition, kinetic calculation using reaction information obtained from the previous chamber studies was performed to estimate concentrations of NDMA and DMN that might be produced from the atmospheric reactions. It was concluded that (1) the atmospheric reactions contributed to the concentrations of NDMA more than they did for those of DMN, (2) the contribution of atmospheric reactions to the concentrations of NDMA and DMN was significant due to high NO2 concentrations in winter, and (3) primary emissions predominantly affected the ambient concentrations of NDMA and DMN in spring, summer, and autumn.

Comparative study of two isothiazolinone biocides, 1,2-benzisothiazolin-3-one (BIT) and 4,5-dichloro-2-n-octyl-isothiazolin-3-one (DCOIT), on barrier function and mitochondrial bioenergetics using murine brain endothelial cell line (bEND.3).

Isothiazolinone (IT) biocides are potent antibacterial substances used as preservatives and disinfectants. These biocides exert differing biocidal effects and display environmental stability based upon chemical structure. In agreement with our recent study reporting that 2-n-octyl-4-isothiazolin-3-one (OIT) induced dysfunction of the blood-brain barrier (BBB), the potential adverse health effects of two IT biocides 1,2-benzisothiazolin-3-one (BIT) and 4,5-dichloro-2-n-octyl-isothiazolin-3-one (DCOIT) were compared using brain endothelial cells (ECs) derived from murine brain endothelial cell line (bEND.3). BIT possesses an unchlorinated IT ring structure and used as a preservative in cleaning products. DCOIT contains a chlorinated IT ring structure and employed as an antifouling agent in paints. Data demonstrated that DCOIT altered cellular metabolism at a lower concentration than BIT. Both BIT and DCOIT increased reactive oxygen species (ROS) generation at the mitochondrial and cellular levels. However, the effect of DCOIT on glutathione (GSH) levels appeared to be greater than BIT. While mitochondrial membrane potential (MMP) was decreased in both BIT- and DCOIT-exposed cells, direct disturbance in mitochondrial bioenergetic flux was only observed in BIT-treated ECs. Taken together, IT biocides produced toxicity in brain EC and barrier dysfunction, but at different concentration ranges suggesting distinct differing mechanisms related to chemical structure.

Using knowledge translation to establish a model of hospital-based early supported community reintegration for stroke patients in South Korea.

BACKGROUND: In 2019, the South Korean government started designating rehabilitation medical institutions to facilitate the early return of patients with stroke (PWS) to their communities after discharge. However, a detailed operating model has not yet been suggested. We aimed to develop a hospital-based early supported community reintegration model for PWS that is suitable for South Korea based on knowledge translation in cooperation with clinical experts and PWS. METHODS: Clinical experts (n = 13) and PWS (n = 20) collaboratively participated in the process of developing the early supported community reintegration model at a national hospital in South Korea, using the following phases of the knowledge-to-action cycle: (1) identifying knowledge, (2) adapting the knowledge to the local situation, (3) assessing barriers and facilitators to local use of knowledge, and (4) tailoring and developing the program. Barriers and facilitators to local use of knowledge were assessed multidimensionally at the individual, interpersonal, organizational, and community level based on the social-ecological model. Literature reviews, workshops, individual and group interviews, and group meetings using nominal group technique were conducted in each phase of the knowledge-to-action cycle. RESULTS: Each phase of the knowledge-to-action cycle for developing the early supported community reintegration model and a newly developed model including the following components were reported: (1) revision of strategies of organizations related to community reintegration support, (2) establishment of a multidepartmental and multidisciplinary community reintegration support system, (3) standardization of patient-centered multidisciplinary goal setting, (4) multidimensional classification of community reintegration support areas, and (5) development of guidelines for a tailored community reintegration support program. CONCLUSIONS: We designed a hospital-based multidimensional and multidisciplinary early supported community reintegration model that comprehensively included several elements of community rehabilitation in connection with hospitals and communities, taking into account the South Korean situation of lacking community rehabilitation infrastructure. In developing a guideline for a tailored community reintegration support program, we attempted to take into consideration various situations faced by PWS in South Korea, which is in a transitional stage for community rehabilitation. It is expected that this early supported community reintegration model can be referenced in other countries that are in a transitional stage of community rehabilitation.

A Commonly Used Biocide 2-N-octyl-4-isothiazolin-3-oneInduces Blood-Brain Barrier Dysfunction via Cellular Thiol Modification and Mitochondrial Damage.

Isothiazolinone (IT) biocides are potent antibacterial substances commonly used as preservatives or disinfectants, and 2-n-Octyl-4-isothiazolin-3-one (OIT; octhilinone) is a common IT biocide that is present in leather products, glue, paints, and cleaning products. Although humans are exposed to OIT through personal and industrial use, the potentially deleterious effects of OIT on human health are still unknown. To investigate the effects of OIT on the vascular system, which is continuously exposed to xenobiotics through systemic circulation, we treated brain endothelial cells with OIT. OIT treatment significantly activated caspase-3-mediated apoptosis and reduced the bioenergetic function of mitochondria in a bEnd.3 cell-based in vitro blood-brain barrier (BBB) model. Interestingly, OIT significantly altered the thiol redox status, as evidenced by reduced glutathione levels and protein S-nitrosylation. The endothelial barrier function of bEnd.3 cells was significantly impaired by OIT treatment. OIT affected mitochondrial dynamics through mitophagy and altered mitochondrial morphology in bEnd.3 cells. N-acetyl cysteine significantly reversed the effects of OIT on the metabolic capacity and endothelial function of bEnd.3 cells. Taken together, we demonstrated that the alteration of the thiol redox status and mitochondrial damage contributed to OIT-induced BBB dysfunction, and we hope that our findings will improve our understanding of the potential hazardous health effects of IT biocides.

Carnosine Protects against Cerebral Ischemic Injury by Inhibiting Matrix-Metalloproteinases.

Stroke is one of the leading causes of death and disability worldwide. However, treatment options for ischemic stroke remain limited. Matrix-metalloproteinases (MMPs) contribute to brain damage during ischemic strokes by disrupting the blood-brain barrier (BBB) and causing brain edemas. Carnosine, an endogenous dipeptide, was found by us and others to be protective against ischemic brain injury. In this study, we investigated whether carnosine influences MMP activity. Brain MMP levels and activity were measured by gelatin zymography after permanent occlusion of the middle cerebral artery (pMCAO) in rats and in vitro enzyme assays. Carnosine significantly reduced infarct volume and edema. Gelatin zymography and in vitro enzyme assays showed that carnosine inhibited brain MMPs. We showed that carnosine inhibited both MMP-2 and MMP-9 activity by chelating zinc. Carnosine also reduced the ischemia-mediated degradation of the tight junction proteins that comprise the BBB. In summary, our findings show that carnosine inhibits MMP activity by chelating zinc, an essential MMP co-factor, resulting in the reduction of edema and brain injury. We believe that our findings shed new light on the neuroprotective mechanism of carnosine against ischemic brain damage.

Establishment of 3D Neuro-Organoids Derived from Pig Embryonic Stem-Like Cells.

Although the human brain would be an ideal model for studying human neuropathology, it is difficult to perform in vitro culture of human brain cells from genetically engineered healthy or diseased brain tissue. Therefore, a suitable model for studying the molecular mechanisms responsible for neurological diseases that can appropriately mimic the human brain is needed. Somatic cell nuclear transfer (SCNT) was performed using an established porcine Yucatan EGFP cell line and whole seeding was performed using SCNT blastocysts. Two Yucatan EGFP porcine embryonic stem-like cell (pESLC) lines were established. These pESLC lines were then used to establish an in vitro neuro-organoids. Aggregates were cultured in vitro until 61 or 102 days after neural induction, neural patterning, and neural expansion. The neuro-organoids were sampled at each step and the expression of the dopaminergic neuronal marker (TH) and mature neuronal marker (MAP2) was confirmed by reverse transcription-PCR. Expression of the neural stem cell marker (PAX6), neural precursor markers (S100 and SOX2), and early neural markers (MAP2 and Nestin) were confirmed by immunofluorescence staining. In conclusion, we successfully established neuro-organoids derived from pESLCs in vitro. This protocol can be used as a tool to develop in vitro models for drug development, patient-specific chemotherapy, and human central nervous system disease studies.