SON is an essential RNA splicing factor promoting ErbB2 and ErbB3 expression in breast cancer.

BACKGROUND: In breast cancer, ErbB receptors play a critical role, and overcoming drug resistance remains a major challenge in the clinic. However, intricate regulatory mechanisms of ErbB family genes are poorly understood. Here, we demonstrate SON as an ErbB-regulatory splicing factor and a novel therapeutic target for ErbB-positive breast cancer. METHODS: SON and ErbB expression analyses using public database, patient tissue microarray, and cell lines were performed. SON knockdown assessed its impact on cell proliferation, apoptosis, kinase phosphorylation, RNA splicing, and in vivo tumour growth. RNA immunoprecipitation was performed to measure SON binding. RESULTS: SON is highly expressed in ErbB2-positive breast cancer patient samples, inversely correlating with patient survival. SON knockdown induced intron retention in selective splice sites within ErbB2 and ErbB3 transcripts, impairing effective RNA splicing and reducing protein expression. SON disruption suppressed downstream kinase signalling of ErbB2/3, including the Akt, p38, and JNK pathways, with increased vulnerability in ErbB2-positive breast cancer cells compared to ErbB2-negative cells. SON silencing in ErbB2-positive breast cancer xenografts led to tumour regression in vivo. CONCLUSION: We identified SON as a novel RNA splicing factor that plays a critical role in regulating ErbB2/3 expression, suggesting SON is an ideal therapeutic target in ErbB2-positive breast cancers.

Electromyography-Triggered Constraint-Induced Movement Cycling Therapy for Enhancing Motor Function in Chronic Stroke Patients: A Randomized Controlled Trial.

This single-blind randomized controlled trial investigated the effectiveness of surface electromyography (sEMG)-triggered constraint-induced movement cycling therapy (CIMCT) in improving balance, lower extremity strength, and activities of daily living in patients with chronic stroke. The participants included patients with chronic stroke-induced hemiplegia who had been diagnosed for more than 6 months, with a minimum score of 24 points on the Mini-Mental State Examination and above level 3 on the Brunnstrom stages. The trial lasted 4 weeks and participants were divided into a CIMCT group and a general cycling training (GCT) group. The CIMCT group (n = 20) used an sEMG-triggered constrained-induced movement therapy device, whereas the GCT group (n = 19) used a standard stationary bicycle. The primary outcome measures showed a significant increase in muscle strength on the affected side in the CIMCT group, as assessed by a manual muscle tester (p < 0.05), with a large effect size (d = 1.86), while no meaningful improvement was observed in the GCT group. Both groups demonstrated significant improvements in dynamic balance, as measured by the Timed Up and Go (TUG) test (p < 0.05), with the CIMCT group showing superior results compared to the GCT group, reflected by a large effect size (d = 0.96). Additionally, both groups showed significant improvements in balance as assessed by the Berg Balance Scale (BBS) and the Functional Reach Test (FRT). The CIMCT group exhibited more pronounced improvements than the GCT group, with large effect sizes of 0.83 for the BBS and 1.25 for the FRT. The secondary outcome measures revealed significant improvements in activities of daily living in both groups, as assessed by the modified Barthel index (MBI), with the CIMCT group achieving a substantial improvement (p < 0.05), accompanied by a large effect size (d = 0.87). This study concludes that sEMG-triggered CIMCT effectively improved muscle strength, postural balance, and activities of daily living in patients with chronic stroke.

Double dissociation of visuomotor interaction mediated by visual feedback during continuous de novo motor learning.

While the sensorimotor cortices are central neural substrates for motor control and learning, how the interaction between their subregions with visual cortices contributes to acquiring de novo visuomotor skills is poorly understood. We design a continuous visuomotor task in fMRI where participants control a cursor using their fingers while learning an arbitrary finger-to-cursor mapping. To investigate visuomotor interaction in the de novo motor task, we manipulate visual feedback of a cursor such that they learn to control using fingers under two alternating conditions: online cursor feedback is available or unavailable except when a target is reached. As a result, we find double dissociation of fMRI activity in subregions of the sensorimotor and visual cortices. Specifically, motor and late visual cortices are more active with online cursor feedback, and somatosensory and early visual cortices are more active without online cursor feedback. We also find a significant reduction in functional connectivity between somatosensory cortices and early visual cortices, which is highly correlated with performance improvement. These findings support the distinct interaction between subregions of sensorimotor cortices and visual cortices, while the connectivity analysis highlights the critical role of somatosensory cortices during de novo motor learning.

Predictive value of left atrial volumes assessed using real-time three-dimensional echocardiography for pulmonary hypertension in dogs with myxomatous mitral valve disease.

Introduction: Left atrial volume (LAV) obtained using real-time three-dimensional echocardiography (RT3DE) is an independent predictor of post-capillary pulmonary hypertension (PH) in humans; however, no studies have investigated LAV obtained using RT3DE as a predictor of post-capillary PH in dogs with myxomatous mitral valve disease (MMVD). Therefore, we aimed to evaluate the clinical applicability of LAV obtained using RT3DE compared to that obtained using two-dimensional echocardiography (2DE) in dogs with MMVD, with or without PH. Methods: Medical records and echocardiographic images of 237 privately owned dogs with naturally occurring MMVD with or without PH were retrospectively reviewed. A total of 49 privately owned dogs with naturally occurring MMVD, with or without PH, were finally included (35 MMVD without PH, 14 MMVD with PH). The LAV and left ventricular volumes were obtained using 2DE and RT3DE. Echocardiographic parameters were analyzed to identify independent predictors of post-capillary PH. Results: We found that the left atrial and left ventricular volumes obtained using 2DE and RT3DE indexed to body weight and several 2DE-derived variables were univariately associated with post-capillary PH. Furthermore, multivariable logistic regression analysis revealed that the RT3DE minimum LAV indexed to body weight (LAVi min) was the only significant independent predictor of post-capillary PH (odds ratio, 12.86; 95% confidence interval [CI], 2.40-68.99; p = 0.003), with the highest area under the curve value of 0.86 (95% CI, 0.75-0.96; p < 0.001). Discussion: In conclusion, LAV indexed to body weight obtained using 2DE and RT3DE, can be a useful predictor of post-capillary PH in dogs with MMVD. In particular, the RT3DE LAVi min was observed to be the strongest predictor of post-capillary PH.

Association between Gut Microbiota and Metabolic Health and Obesity Status in Cats.

Obesity is a major public health concern in both humans and animals, leading to several metabolic complications. Recent human studies have classified obesity into two phenotypes, metabolically healthy (MHO) and metabolically unhealthy (MUO) obesity based on cardiovascular and metabolic risk factors. MHO cases lack these risk factors and are protected from metabolic complications of obesity, whereas MUO cases exhibit the opposite characteristics. Moreover, recent studies have highlighted the possible role of the gut microbiome in determining metabolic health of obese individuals. However, studies on the association between the gut microbiome and obesity and metabolic abnormalities in cats are limited. Therefore, we aimed to examine the association between metabolic health phenotypes and gut microbiota composition and diversity in obese cats. We investigated hormone and serum biochemistry parameters and composition of the gut microbiota in non-obese (NO), MHO, and MUO groups. The abundances of Bifidobacteriaceae, Coriobacteriaceae, and Veillonellaceae were significantly higher in the obese versus NO group, showing a positive correlation with body mass index. The abundance of Ruminococcaceae was significantly higher in the MUO versus NO group, showing a positive correlation with triglyceride and total cholesterol levels. However, obesity, whether MHO or MUO, had only a minimal impact on fecal microbiota. Therefore, further studies are warranted to investigate whether gut microbiota could be a beneficial tool for the treatment or management of this condition.

Longitudinal proteome-wide antibody profiling in Marburg virus survivors identifies wing domain immunogen for vaccine design.

Limited knowledge exists on the quality of polyclonal antibody responses generated following Marburg virus (MARV) infection and its evolution in survivors. In this study, we evaluate MARV proteome-wide antibody repertoire longitudinally in convalescent phase approximately every six months for five years following MARV infection in ten human survivors. Differential kinetics were observed for IgM vs IgG vs IgA epitope diversity, antibody binding, antibody affinity maturation and Fc-receptor interaction to MARV proteins. Durability of MARV-neutralizing antibodies is low in survivors. MARV infection induces a diverse epitope repertoire with predominance against GP, VP40, VP30 and VP24 that persisted up to 5 years post-exposure. However, the IgM and IgA repertoire declines over time. Within MARV-GP, IgG recognize antigenic sites predominantly in the amino-terminus, wing domain and GP2-heptad repeat. Interestingly, MARV infection generates robust durable FcɣRI, FcɣRIIA and FcɣRIIIA IgG-Fc receptor interactions. Immunization with immunodominant MARV epitopes reveals conserved wing region between GP1 and GP2, induces neutralizing antibodies against MARV. These findings demonstrate that MARV infection generates a diverse, long-lasting, non-neutralizing, IgG antibody repertoire that perturbs disease by FcɣR activity. This information, along with discovery of neutralizing immunogen in wing domain, could aid in development of effective therapeutics and vaccines against Marburg virus.

The factors related to depressive symptoms in urban older adults in South Korea: a study based on the Seoul Aging Survey.

PURPOSE: This study aims to comprehensively examine the determinants of depression among urban older adults in Seoul, utilizing the social-ecological model to address multifaceted influences. METHODS: A comprehensive analysis was conducted using data from the 2022 Seoul Aging Survey, which surveyed 2,914 individuals aged 65 and above. A multiple logistic regression analysis was performed to identify factors contributing to depressive symptoms, including socio-demographic characteristics, health behaviors and status, research accessibility, social support, and environmental influences. RESULTS: This study revealed that poor subjective health (OR = 1.47) and the presence of multiple chronic diseases (OR = 1.59) significantly increased the risk of depressive symptoms among urban older adults. From a social support standpoint, living alone was associated with a higher risk of depression (OR = 1.66), low food security (OR = 2.56), and low digital competency (OR = 2.70) were all significant predictors of depressive symptoms. Additionally, a lack of engagement with cultural facilities (OR = 2.15) was identified as a critical environmental factor contributing to depression. CONCLUSIONS: The findings underscore the need for comprehensive policy and practical interventions aimed at preventing chronic disease, enhancing social support networks, improving digital literacy, ensuring food security, and expanding access to healthcare and cultural facilities. Such measures are crucial in mitigating depression among urban older adults, thereby enhancing their overall well-being and quality of life.

Attitudes and experiences of traditional Korean medicine practitioners in cases of traffic accidents: A qualitative study.

Objective: The surge in vehicles has escalated traffic volume, leading to an upswing in traffic accidents and subsequent disorders. Complex symptoms often characterize post-traumatic syndrome from these accidents. Traditional Korean medicine (TKM), increasingly used in car insurance, forms a substantial part of treatment costs. However, the current system lacks explicit fee guidelines and approval criteria for non-reimbursable TKM procedures, relying heavily on practitioners' judgment without robust evidence-based decision-making. This scenario raises concerns about treatment appropriateness and transparency. We aim to explore physicians' perspectives on utilizing TKM in emergency medicine, their participation sentiments, and their session selection process post-traffic accident. Methods: We collected TKM practitioners' opinions regarding their role in clinical environment and involvement in treating patients after traffic accidents. The need for comprehensive and standardized protocols for the diagnosis, treatment, management, and prognosis of patients with post-traumatic syndrome is evident. Additionally, improvements that facilitate rational decision-making by medical consumers and protect the treatment rights of healthcare providers are necessary. Results has emphasized the importance of evidence-based decision-making, establishing appropriate fee structures and detailed criteria for non-reimbursable TKM-based procedures, and enhancing regulations for the reliability and transparency of TKM-based treatments in the context of car insurance. Results and conclusions: The perspective of healthcare providers directly involved in TKM-based treatments must be considered to maintain a sustainable vehicular insurance system, transcending administrative policy discourse. We highlighted the challenges and potential solutions for improving the effectiveness and appropriateness of TKM-based treatments in the context of car insurance.

Direct Observation and Analysis of Low-Energy Magnons with Raman Spectroscopy in Atomically Thin NiPS3.

van der Waals (vdW) magnets have rapidly emerged as a fertile playground for fundamental physics and exciting applications. Despite the impressive developments over the past few years, technical limitations pose a severe challenge to many other potential breakthroughs. High on the list is the lack of suitable experimental tools for studying spin dynamics on atomically thin samples. Here, Raman scattering techniques are employed to directly observe the low-lying magnon (∼1 meV) even in bilayer NiPS3. The advantage is that it offers excellent energy resolutions far better on low-energy sides than most inelastic neutron spectrometers can offer. More importantly, with appropriate theoretical analysis, the polarization dependence of the Raman scattering by those low-lying magnons also provides otherwise hidden information on the dominant spin-exchange scattering paths for different magnons. By comparing with high-resolution inelastic neutron scattering data, these low-energy Raman modes are confirmed to be indeed of magnon origin. Because of the different scattering mechanisms involved in inelastic neutron and Raman scattering, this information is fundamental in pinning down the final spin Hamiltonian. This work demonstrates the capability of Raman spectroscopy to probe the genuine two-dimensional spin dynamics in atomically thin vdW magnets, which can provide insights that are obscured in bulk spin dynamics.

Dual-Salts Localized High-Concentration Electrolyte for Li- and Mn-Rich High-Voltage Cathodes in Lithium Metal Batteries.

Limited electrochemical stability windows of conventional carbonate-based electrolytes pose a challenge to support the Lithium (Li)- and manganese (Mn)-rich (LMR) high-voltage cathodes in rechargeable Li-metal batteries (LMBs). To address this issue, a novel localized high-concentration electrolyte (LHCE) composition incorporating LiPF6 and LiTFSI as dual-salts (D-LHCE), tailored for high-voltage (>4.6 Vvs.Li) operation of LMR cathodes in LMBs is introduced. 7Li nuclear magnetic resonance and Raman spectroscopy revealed the characteristics of the solvation structure of D-LHCE. The addition of LiPF6 provides stable Al-current-collector passivation while the addition of LiTFSI improves the stability of D-LHCE by producing a more robust cathode-electrolyte interphase (CEI) on LMR cathode and solid-electrolyte interphase (SEI) on Li-metal anode. As a result, LMR/Li cell, using the D-LHCE, achieved 72.5% capacity retention after 300 cycles, a significant improvement compared to the conventional electrolyte (21.9% after 100 cycles). The stabilities of LMR CEI and Li-metal SEI are systematically analyzed through combined applications of electrochemical impedance spectroscopy and distribution of relaxation times techniques. The results present that D-LHCE concept represents an effective strategy for designing next-generation electrolytes for high-energy and high-voltage LMB cells.

ECMO is associated with decreased hospital mortality in COVID-19 ARDS.

This study determined whether compared to conventional mechanical ventilation (MV), extracorporeal membrane oxygenation (ECMO) is associated with decreased hospital mortality or fibrotic changes in patients with COVID-19 acute respiratory distress syndrome. A cohort of 72 patients treated with ECMO and 390 with conventional MV were analyzed (February 2020-December 2021). A target trial was emulated comparing the treatment strategies of initiating ECMO vs no ECMO within 7 days of MV in patients with a PaO2/FiO2 < 80 or a PaCO2 ≥ 60 mmHg. A total of 222 patients met the eligibility criteria for the emulated trial, among whom 42 initiated ECMO. ECMO was associated with a lower risk of hospital mortality (hazard ratio [HR], 0.56; 95% confidence interval [CI] 0.36-0.96). The risk was lower in patients who were younger (age < 70 years), had less comorbidities (Charlson comorbidity index < 2), underwent prone positioning before ECMO, and had driving pressures ≥ 15 cmH2O at inclusion. Furthermore, ECMO was associated with a lower risk of fibrotic changes (HR, 0.30; 95% CI 0.11-0.70). However, the finding was limited due to relatively small number of patients and differences in observability between the ECMO and conventional MV groups.

Hypertrophic Osteopathy Concurrent with an Aberrant Right Subclavian Artery in a Dog.

A 13-year-old spayed female cocker spaniel was presented with a 2-month history of swelling in several digits and intermittent hindlimb lameness. Radiographs revealed marked soft-tissue swelling and periosteal new bone formation without cortical bone destruction, characteristic of hypertrophic osteopathy (HO), in the distal parts of all extremities except for the right forelimb. However, no notable findings were detected in thoracic radiographs. An ultrasonography indicated cranial bladder wall thickening, which resolved following antibiotic therapy. Computed tomographic angiography identified a potential underlying cause as an aberrant right subclavian artery (ARSA) originating from the aortic arch, compressing the esophagus and causing mild esophageal cranial dilation to the aberrant vessel. No other intrathoracic or neoplastic lesions were observed. Gastrointestinal symptoms, such as regurgitation, were absent. Although an ARSA was likely the cause of HO, surgical correction was declined by the owner. To the best of our knowledge, this is the first reported case of HO concurrent with ARSA in dogs.

Adapt-cMolGPT: A Conditional Generative Pre-Trained Transformer with Adapter-Based Fine-Tuning for Target-Specific Molecular Generation.

Small-molecule drug design aims to generate compounds that target specific proteins, playing a crucial role in the early stages of drug discovery. Recently, research has emerged that utilizes the GPT model, which has achieved significant success in various fields to generate molecular compounds. However, due to the persistent challenge of small datasets in the pharmaceutical field, there has been some degradation in the performance of generating target-specific compounds. To address this issue, we propose an enhanced target-specific drug generation model, Adapt-cMolGPT, which modifies molecular representation and optimizes the fine-tuning process. In particular, we introduce a new fine-tuning method that incorporates an adapter module into a pre-trained base model and alternates weight updates by sections. We evaluated the proposed model through multiple experiments and demonstrated performance improvements compared to previous models. In the experimental results, Adapt-cMolGPT generated a greater number of novel and valid compounds compared to other models, with these generated compounds exhibiting properties similar to those of real molecular data. These results indicate that our proposed method is highly effective in designing drugs targeting specific proteins.

Establishment and characterization of six canine hepatocellular carcinoma cell lines.

Background: Hepatocellular carcinoma (HCC) is the most common malignant liver tumor in dogs. Although surgical resection is a major treatment option for canine HCC, there are no distinct strategies for unresectable tumor subtypes or adjuvant chemotherapy for tumors with positive margins. We aimed to establish and characterize novel HCC cell lines from canine patients. Methods: The cellular morphology, general growth features and tumorigenicity of the established cell lines were evaluated. We also examined the sensitivity of the cell lines to multi-target tyrosine kinase inhibitors (TKIs). Results: We established novel canine HCC cell lines from hepatic tumors and an additional kidney tumor of six canine patients. All cell lines showed colony forming and migratory ability. KU-cHCC-001 and KU-cHCC-001-Kidney, two cell lines exhibiting high epithelial-mesenchymal transition characteristics, showed tumorigenicity in xenografted mice. Toceranib, a veterinary TKI that targets vascular endothelial growth factor (VEGFR)/platelet-derived growth factor receptor (PDGFR)/c-kit, effectively inhibited the mitogen-activated protein kinase pathway and induced apoptosis. The established canine HCC cell lines showed greater sensitivity to toceranib than to sorafenib, a first-line treatment for human HCC targeting RAF/VEGFR/PDGFR. Sorafenib showed improved anti-tumor effects when co-treated with SCH772984, an extracellular signal-regulated kinase inhibitor. Conclusion: Our study suggests new therapeutic strategies for canine HCC, and these cell lines are valuable research materials for understanding HCC tumor biology in both humans and dogs.

Comparative analysis and classification of highly divergent mouse rDNA units based on their intergenic spacer (IGS) variability.

Ribosomal DNA (rDNA) repeat units are organized into tandem clusters in eukaryotic cells. In mice, these clusters are located on at least eight chromosomes and show extensive variation in the number of repeats between mouse genomes. To analyze intra- and inter-genomic variation of mouse rDNA repeats, we selectively isolated 25 individual rDNA units using Transformation-Associated Recombination (TAR) cloning. Long-read sequencing and subsequent comparative sequence analysis revealed that each full-length unit comprises an intergenic spacer (IGS) and a ∼13.4 kb long transcribed region encoding the three rRNAs, but with substantial variability in rDNA unit size, ranging from ∼35 to ∼46 kb. Within the transcribed regions of rDNA units, we found 209 variants, 70 of which are in external transcribed spacers (ETSs); but the rDNA size differences are driven primarily by IGS size heterogeneity, due to indels containing repetitive elements and some functional signals such as enhancers. Further evolutionary analysis categorized rDNA units into distinct clusters with characteristic IGS lengths; numbers of enhancers; and presence/absence of two common SNPs in promoter regions, one of which is located within promoter (p)RNA and may influence pRNA folding stability. These characteristic features of IGSs also correlated significantly with 5'ETS variant patterns described previously and associated with differential expression of rDNA units. Our results suggest that variant rDNA units are differentially regulated and open a route to investigate the role of rDNA variation on nucleolar formation and possible associations with pathology.

Chronological distribution and potential sources of persistent toxic substances in soils from the glacier foreland of Midtre Lovénbreen, Svalbard.

Despite its reputation as one of the cleanest regions globally, recent studies have identified the presence of various persistent toxic substances (PTSs) in the environmental matrices collected from Svalbard. This study investigated the chronological distribution and potential sources of 81 PTSs in soils from the glacier foreland of Midtre Lovénbreen. Soil samples (n = 45) were categorized by age based on exposure to the atmosphere due to glacier retreat in July 2014 into five age groups: 80-100 years (n = 7), 60-80 years (n = 12), 40-60 years (n = 16), 20-40 years (n = 7), and <20 years (n = 3). Concentrations of polychlorinated biphenyls (PCBs, n = 32) in soils varied with age, ranging from 0.29 to 0.74 ng g-1 dw. In addition, the concentrations of polycyclic aromatic hydrocarbons (PAHs, n = 28), perylene, and alkyl-PAHs (n = 20) in soils ranged from 21 to 80 ng g-1 dw, 2.9-62 ng g-1 dw, and 73-420 ng g-1 dw, respectively. The concentrations of PTSs were observed to be greater in older soils. Principal component analysis revealed that PCBs in soils originated from various product sources. Positive matrix factorization modeling estimated the association of PAHs in soils with potential origins, such as diesel emissions, petroleum and coal combustion, and coal. Potential sources of PAHs were mainly coal in younger soils and diesel emissions and petroleum combustion in older soils. Alkyl-PAH compositions in the soil were similar to those of bituminous coal, with a noteworthy degree of weathering observed in older soils. The accumulation rate and flux of PTSs in soils exhibited compound-specific patterns, reflecting factors such as long-range transport, fate, origin, and recent inputs. These findings can serve as baseline data for protecting and preserving polar environments.

Successful post-incomplete resection management of gastrointestinal stromal tumor using imatinib based on adenosine triphosphate-based tumor sensitivity assay in a dog.

Gastrointestinal stromal tumors arising from gastric cardia are uncommon in dogs. A few studies have shown the effectiveness of tyrosine kinase inhibitors in the treatment of canine gastrointestinal stromal tumors, but no standardized protocols are currently available. An 11-year-old spayed female Maltese dog was diagnosed with a gastrointestinal stromal tumor using histopathological and immunohistochemical analyses. An adenosine triphosphate-based tumor chemosensitivity assay revealed that imatinib at lower concentrations had a stronger inhibitory effect than toceranib. Based on the results of the assay, the dog was treated with imatinib after surgery. After 28 mo of therapy, there was no recurrence of the tumor. Key clinical message: Adenosine triphosphate-based tumor chemosensitivity assays may help clinicians to select appropriate postoperative chemotherapeutic drugs for incompletely resected gastrointestinal stromal tumors in dogs.

Gestion réussie à la suite d'une résection incomplète d'une tumeur stromale gastro-intestinale à l'aide de l'imatinib basée sur un test de sensibilité tumorale à base d'adénosine triphosphate chez un chien. Les tumeurs stromales gastro-intestinales résultant du cardia gastrique sont rares chez le chien. Quelques études ont montré l'efficacité des inhibiteurs de la tyrosine kinase dans le traitement des tumeurs stromales gastrointestinales canines, mais aucun protocole standardisé n'est actuellement disponible. Une chienne maltaise stérilisée de 11 ans a reçu un diagnostic de tumeur stromale gastro-intestinale à l'aide d'analyses histopathologiques et immunohistochimiques. Un test de chimiosensibilité tumorale à base d'adénosine triphosphate a révélé que l'imatinib à des concentrations plus faibles avait un effet inhibiteur plus fort que le tocéranib. Sur la base des résultats du test, le chien a été traité avec de l'imatinib après l'opération. Après 28 mois de traitement, il n'y a eu aucune récidive de la tumeur.Message clinique clé :Les tests de chimiosensibilité tumorale à base d'adénosine triphosphate peuvent aider les cliniciens à sélectionner les médicaments chimiothérapeutiques postopératoires appropriés pour les tumeurs stromales gastro-intestinales incomplètement réséquées chez le chien.(Traduit par Dr Serge Messier).

Contribution of carbonate-derived dissolved inorganic carbon into autochthonous particulate organic carbon in two small temperate Korean rivers (Geum and Seomjin).

In this study, we estimated the contributions of carbonate mineral weathering to dissolved inorganic carbon (DIC) and carbonate-derived DIC to autochthonous particulate organic carbon (POC) in two temperate Korean rivers. We combined stoichiometric and stable carbon isotopic approaches to calculate the contribution of autochthonous POC, considering diverse riverine DIC sources. We collected surface water samples from May 2016 to May 2018 and analyzed the major ion composition of rivers along with the concentrations and stable carbon isotopes of DIC. Our estimates showed that the relative abundances of carbonate mineral weathering (0.41 ± 0.11 in the Geum River and 0.43 ± 0.07 in the Seomjin River) were only slightly lower than those of silicate mineral weathering (0.59 ± 0.1 in the Geum River and 0.57 ± 0.07 in the Seomjin River). The resulting percentage contributions of DIC derived from the carbonate mineral weathering to riverine autochthonous POC, if we consider the additional DIC sources of atmospheric and soil-derived CO2, were 10 ± 3 % in the Geum River and 2 ± 1 % in the Seomjin River. The calculated annual fluxes of carbonate-derived DIC for 2016-2018 were 23.2 ± 0.3 Gg C yr-1 in the Geum River and 1.1 ± 0.4 Gg C yr-1 in the Seomjin River. Moreover, the calculated annual fluxes of carbonate-derived POC were 3.6 ± 0.5 Gg C yr-1 in the Geum River and 0.1 ± 0.7 Gg C yr-1 in the Seomjin River. Accordingly, our study provides the first insight into the contribution of carbonate-derived DIC to riverine autochthonous POC in small temperate Korean river systems, dominated by silicate rocks.

Guidelines for Manufacturing and Application of Organoids: Lung.

The objective of standard guideline for utilization of human lung organoids is to provide the basic guidelines required for the manufacture, culture, and quality control of the lung organoids for use in non-clinical efficacy and inhalation toxicity assessments of the respiratory system. As a first step towards the utilization of human lung organoids, the current guideline provides basic, minimal standards that can promote development of alternative testing methods, and can be referenced not only for research, clinical, or commercial uses, but also by experts and researchers at regulatory institutions when assessing safety and efficacy.

LC-MS-MS method for mitragynine and 7-hydroxymitragynine in hair and its application in authentic hair samples of suspected kratom abusers.

Kratom is a natural psychoactive product known primarily in Southeast Asia, including Thailand, Malaysia, etc. It is also known as krathom, kakuam, ithang, thom (Thailand), biak-biak, ketum (Malaysia) and mambog (Philippines) and is sometimes used as an opium substitute. It is stimulant at doses of 1-5 g, analgesic at doses of 5-15 g and euphoric and sedative at doses of >15 g. Mitragynine is the most abundant indole compound in kratom (Mitragyna speciosa) and is metabolized in humans to 7-hydroxymitragynine, the more active metabolite. Adverse effects include seizures, nausea, vomiting, diarrhea, tachycardia, restlessness, tremors, hallucinations and death. There are few studies on the analytical method for the detection of mitragynine and 7-hydroxymitragynine in hair. Therefore, this study proposes a liquid chromatography-tandem mass spectrometry (LC-MS-MS) method for the analysis of kratom in hair. Hair samples were first weighed to ∼10 mg and washed with methanol. Then the washed hair samples were cut into pieces and incubated in methanol with stirring and heating (16 h/38℃). Extracts were then analyzed by LC-MS-MS. This method was validated by determining the limit of detection (LOD), limit of quantification, linearity, intra- and inter-day accuracy and precision, recovery and matrix effects. The intra- and inter-day precision (CV%) and accuracy (bias%) were within ±20%, which was considered acceptable. Using this newly developed LC-MS-MS method, the simultaneous detection of mitragynine and 7-hydroxymitragynine in six authentic hair samples was achieved to provide the direct evidence of kratom use in the past. Mitragynine concentrations ranged from 16.0 to 2,067 pg/mg (mean 905.3 pg/mg), and 7-hydroxymitragynine concentrations ranged from 0.34 to 15 pg/mg (mean 7.4 pg/mg) in six authentic hair samples from kratom abusers. This may be due to the higher sensitivity of the LOD in this study, with values of 0.05 pg/mg for mitragynine and 0.2 pg/mg for 7-hydroxymitragynine in hair.