The reprotoxic adverse side effects of neurogenic and neuroprotective drugs: current use of human organoid modeling as a potential alternative to preclinical models.

The management of neurological disorders heavily relies on neurotherapeutic drugs, but notable concerns exist regarding their possible negative effects on reproductive health. Traditional preclinical models often fail to accurately predict reprotoxicity, highlighting the need for more physiologically relevant systems. Organoid models represent a promising approach for concurrently studying neurotoxicity and reprotoxicity, providing insights into the complex interplay between neurotherapeutic drugs and reproductive systems. Herein, we have examined the molecular mechanisms underlying neurotherapeutic drug-induced reprotoxicity and discussed experimental findings from case studies. Additionally, we explore the utility of organoid models in elucidating the reproductive complications of neurodrug exposure. Have discussed the principles of organoid models, highlighting their ability to recapitulate neurodevelopmental processes and simulate drug-induced toxicity in a controlled environment. Challenges and future perspectives in the field have been addressed with a focus on advancing organoid technologies to improve reprotoxicity assessment and enhance drug safety screening. This review underscores the importance of organoid models in unraveling the complex relationship between neurotherapeutic drugs and reproductive health.

Dried blood spot sampling coupled with liquid chromatography-tandem mass for simultaneous quantitative analysis of multiple cardiovascular drugs.

Dried Blood Spots (DBS) revolutionize therapeutic drug monitoring using LC-MS for the precise quantification of cardiovascular drugs (CDs), enabling personalized treatment adapted to patient-specific pharmacokinetics with minimal invasiveness. This study aims to achieve simultaneous quantification of eight CDs in DBS, overcoming physicochemical challenges. A two-step protein precipitation method was used for simple and precise sample preparation. The drugs were analyzed using LC-MS/MS in ESI positive-ion mode, showing high sensitivity and linearity, with a correlation coefficient (r2) exceeding 0.999, after being separated on a reversed-phase chromatography by gradient elution of DW-acetonitrile containing 0.1 % formic acid + 2 mM ammonium formate. The validation results indicate good selectivity, with no observed matrix effect and carry-over. The intra- and inter-day accuracy and precision were within 6 % for most drugs, except for digoxin and deslanoside at low therapeutic levels where the variation was within 20 %. Stability tests confirmed suitable DBS handling and storage conditions, indicating drug stability for at least 30 days at room temperature. The analysis of whole spot has demonstrated remarkable precision and reliability in all target drugs. The analysis of 3 mm internal diameter discs, punched in and out of DBS, presumed to contain 3 µL of blood, showed acceptable accuracy for most drugs, with less polar drugs like digoxin and deslanoside showing lower accuracy, indicating a need for further correction due to non-uniform drug distribution. Consequently, the developed LC-MS/MS method enables the quantification of multiple CDs in a single DBS analysis, while suggesting the potential for accuracy-based analysis.

Melatonin and resveratrol alleviate molecular and metabolic toxicity induced by Bisphenol A in endometrial organoids.

Bisphenol A (BPA), a widespread environmental contaminant, poses concerns due to its disruptive effects on physiological functions of the uterine endometrium. In contrast, melatonin (MT) and Resveratrol (RSV) are under scrutiny for their potential protective roles against BPA-induced damage. For the efficacy and ethical concerns in the animal test, endometrial organoids, three-dimensional models mimicking endometrium, serve as crucial tools for unraveling the impact of environmental factors on reproductive health. This study aimed to comprehensively characterize the morphological, molecular and metabolic responses of porcine endometrial organoids to BPA and assess the potential protective effects of MT and RSV. Porcine uteri were prepared, digested with collagenase, mixed with Matrigel, and incubated at 38°C with 5 % CO2. Passaging involved dissociation through trypsin-EDTA treatment and subculturing. The culture medium was refreshed every 2-3 days. To investigate the environmental impact on reproductive health, endometrial organoids were treated with BPA (0.5 µM), MT (with/without BPA at 0.1 µM), and/or RSV (10 µM). Various molecular screening using gene expression, western blotting, immunofluorescence staining, and metabolites profiling were assessed the effects of BPA, MT, and RSV in terms of cell viability, morphology, reproductivity, and metabolism alteration in the endometrial organoids. As expected, BPA induced structural and molecular disruptions in organoids, affecting cytoskeletal proteins, Wnt/ß-catenin signaling, and epithelial/mesenchymal markers. It triggered oxidative stress and apoptotic pathways, altered miRNA expression, and disrupted the endocannabinoid system. The level of glucose, galactose, and essential amino acids were increased or decreased by approximately 1.5-3 times in BPA-treated groups compared to the control groups (p-value < 0.05), indicating metabolic changes. Moreover, MT and RSV treated groups exhibited protective effects, mitigating BPA-induced disruptions across multiple pathways. For the first time, our study models endometrial organoids, advancing understanding of environmental impacts on reproductive health.

Simultaneous quantification of 11 antiepileptic drugs using limited isotope-labeled internal standards in LC-MS/MS: An accuracy assessment.

This study aims to establish an LC-MS/MS method to simultaneously analyze 11 antiepileptic drugs with a particular focus on maintaining accuracy while reducing the number of isotope-labeled internal standards employed for cost-effectiveness. By applying a water/acetonitrile gradient elution containing 0.1 % formic acid and 2 mM ammonium formate as the mobile phase, optimal sensitivity for the target drugs could be obtained in positive ESI mode in LC-MS/MS. After optimizing various extraction techniques, extraction with 70 % acetonitrile was selected as it provided good recoveries (>93 %) for all targets without matrix effects. Accuracies within 3 % were achieved from the combination of six internal standards, while accuracies of 5 % and 10 % were obtained by reducing the number of internal standards to four and two, respectively, for more economical analysis. The accuracy of the established method was maintained in hyperglycemia, hyperlipidemia, and hyperalbuminemia sera, suggesting that it can be successfully applied to individual serum samples with various properties.

The secreted protein Amuc_1409 from Akkermansia muciniphila improves gut health through intestinal stem cell regulation.

Akkermansia muciniphila has received great attention because of its beneficial roles in gut health by regulating gut immunity, promoting intestinal epithelial development, and improving barrier integrity. However, A. muciniphila-derived functional molecules regulating gut health are not well understood. Microbiome-secreted proteins act as key arbitrators of host-microbiome crosstalk through interactions with host cells in the gut and are important for understanding host-microbiome relationships. Herein, we report the biological function of Amuc_1409, a previously uncharacterised A. muciniphila-secreted protein. Amuc_1409 increased intestinal stem cell (ISC) proliferation and regeneration in ex vivo intestinal organoids and in vivo models of radiation- or chemotherapeutic drug-induced intestinal injury and natural aging with male mice. Mechanistically, Amuc_1409 promoted E-cadherin/ß-catenin complex dissociation via interaction with E-cadherin, resulting in the activation of Wnt/ß-catenin signaling. Our results demonstrate that Amuc_1409 plays a crucial role in intestinal homeostasis by regulating ISC activity in an E-cadherin-dependent manner and is a promising biomolecule for improving and maintaining gut health.

Description of Mycolicibacterium arenosum sp. nov. Isolated from Coastal Sand on the Yellow Sea Coast.

A Gram-staining-positive, aerobic, non-spore-forming bacterium was isolated from coastal sand samples from Incheon in the Republic of Korea and designated as strain CAU 1645T. The optimum conditions for growth were observed at 30 °C in growth media containing 1% (w/v) NaCl at pH 9.0. The predominant respiratory quinone was MK-9 and the major fatty acids were C16:0, C17:1 w7c, and summed feature 7. Similarly, the 16S rRNA gene sequence exhibited the highest similarity with Mycolicibacterium bacteremicum DSM 45578T and Mycolicibacterium neoaurum JCM 6365T, both of which exhibited similarity rates of 97.2%. The genomic DNA G+C content was 68.2%. The whole genome of strain CAU 1645T was obtained and annotated with annotation using RAST server. The pan-genome analysis was determined using Prokka, Roary, and Phandango. In the pan-genome analysis, the strain CAU 1645T shared 40 core genes with closely related Mycolicibacterium species, including the AcpM gene, the meromycolate extension acyl carrier protein involved in forming impermeable cell walls in mycobacteria. Therefore, our findings demonstrated that the isolate represents a novel species of the genus Mycolicibacterium, for which we propose the name Mycolicibacterium arenosum sp. nov. The type strain is CAU 1645T (= KCTC 49724T = MCCC 1K07087T).

Generation of porcine endometrial organoids and their use as a model for enhancing embryonic attachment and elongation.

In brief: Porcine endometrial organoids (EOs) were isolated and characterized, revealing distinctive features such as unique extracellular matrix formation, fusion into uterine bud-like structures, and facilitation of embryo elongation. The yield of EOs was significantly enhanced by cryopreservation medium supplemented with the rock inhibitor (Y-27632), resulting in reduced expression of apoptotic mRNAs and microRNAs. Abstract: Endometrial organoids (EOs) are acceptable models for understanding maternal-embryonic cross talk. This study was conducted to generate EOs and optimize their cryopreservation and provide coculture modeling with embryos. The endometrial tissues were used for culturing the organoids inside domes of Matrigel®. To improve the long-term storage of EOs, 10 µM ROCK inhibitor (RI) was added to the cryopreservation medium. Day 7 parthenogenetically activated embryos were cocultured with EOs or EO outgrowths, and embryonic cell numbers and embryo attachment were monitored. Spherical EOs 100-300 µm in size can be retrieved on day 7 of culture, and larger EOs, approximately 1.5 mm in diameter, can be maintained in the Matrigel® dome for 21 days. The nuclear expression of Ki67 indicates that more than 80% of EOs nuclei were proliferative. EOs exhibit unique novel characters such as formation of extracellular matrix and ability for fusion. RI increased the yield and quality of organoids after freezing or thawing. The cell number of cocultured embryos increased five-fold, and the proportion of trophoblast outgrowths increased seven-fold compared with those of control embryos. The embryos cultured with EO-conditioned medium showed a better attachment rate than the other models, and - for the first time - we report embryonic elongation. Immunofluorescence staining of the attached embryos showed CDX2 in the periphery of EOs outgrowths. The 3D assembly and cryopreservation of EOs was optimized, and EO coculture supported embryo attachment, trophoblast outgrowth, and elongation, which would provide a valuable tool for studying the intricate processes involved in porcine embryo implantation.

Simultaneous quantification method for multiple antiviral drugs in serum using isotope dilution liquid chromatography-tandem mass spectrometry.

We describe the simultaneous quantification of six antiviral drugs in serum based on high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). The target drugs-hydroxychloroquine, chloroquine, favipiravir, umifenovir, ritonavir, and lopinavir-were extracted and purified from serum with 75 % v/v methanol as the precipitant reagent. The six analytes were clearly separated within 15 min using gradient elution and mixed-mode stationary phase. The measurement accuracy and precision were assured by adopting isotopes as internal standards. The optimized measurement procedure was strictly validated in linearity, sensitivity, accuracy, and precision. To confirm the robustness of the method in matrix, the method was additionally applied to various types of serum, namely hyperlipidemic and hyperglycemic serum. The method was then applied to assess the stability of the drugs in serum in order to set sample handling and storage guides for laboratory testing. Lastly, the method was implemented in different LC-MS systems to confirm its applicability across similar equipment commonly used in clinical testing laboratories. The overall results show that the optimized protocol is suitable for the accurate, simultaneous quantification of the six antiviral drugs in serum, and it is anticipated to satisfactorily serve as a reference protocol for the analysis of a wide range of other antiviral drugs for drug monitoring with various purposes.

Incident infection risks depending on oral antidiabetic exposure in insulin-treated type 2 diabetes patients.

Dipeptidyl peptidase-4 inhibitors (DPP4is) and sodium glucose cotransporter-2 inhibitors (SGLT2is) have been speculated to have a potential to increase infection risks in type 2 diabetes mellitus (T2DM) patients. We performed a cohort study using the Korean health insurance data to investigate infection risks with each drug class relative to metformin in insulin-treated T2DM patients. After propensity score matching, we included 1,498 and 749 patients in DPP4i + insulin vs metformin + insulin and 300 and 549 patients in SGLT2i + insulin vs metformin + insulin, respectively. In stratified analyses per patient factor, none of the odds ratios (ORs) were associated with a statistical significance across respiratory, genital, and urinary tract infections (UTIs), except that of the male stratum for respiratory infections (OR 0.77, p = 0.04). With regard to SGLT2is, a higher risk of genital infections was analyzed with their use than with metformin therapy (OR 1.76, p = 0.03). In stratified analyses, the OR for genital infections remained significant in the baseline cardiovascular disease stratum (OR 2.29, p = 0.01). No increased UTI risk was detected with SGLT2is compared against metformin. In this study on insulin-receiving T2DM patients, DPP4is were not associated with increased infection risks, whereas SGLT2is led to a higher risk for genital infections, but not for UTIs, relative to metformin.

Predictive modelling for the diagnosis of oral and laryngeal premalignant and malignant lesions using p53 and Ki-67 expression.

Oral and laryngeal epithelial lesions are currently diagnosed using histological criteria based on the World Health Organization (WHO) classification, which can cause interobserver variability. An integrated diagnostic approach based on immunohistochemistry (IHC) would aid in the interpretation of ambiguous histological findings of epithelial lesions. In the present study, IHC was used to evaluate the expression of p53 and Ki-67 in 114 cases of oral and laryngeal epithelial lesions in 104 patients. Logistic regression analysis and decision tree algorithm were employed to develop a scoring system and predictive model for differentiating the epithelial lesions. Cohen's kappa coefficient was used to evaluate interobserver variability, and next-generation sequencing (NGS) and IHC were used to compare TP53 mutation and p53 expression patterns. Two expression patterns for p53, namely, diffuse expression type (pattern HI) and null type (pattern LS), and the pattern HI for Ki-67 were significantly associated with high-grade dysplasia (HGD) or squamous cell carcinoma (SqCC). With an accuracy and area under the receiver operating characteristic curve (AUC) of 84.6% and 0.85, respectively, the scoring system based on p53 and Ki-67 expression patterns classified epithelial lesions into two types: non-dysplasia (ND) or low-grade dysplasia (LGD) and SqCC or HGD. The decision tree model constructed using the p53 and Ki-67 expression patterns classified epithelial lesions into ND, LGD, and group 2, including HGD or SqCC, with an accuracy and AUC of 75% and 0.87, respectively. The integrated diagnosis had a better correlation with near perfect agreement (weighted kappa 0.92, unweighted kappa 0.88). The patterns HI and LS for p53 were confirmed to be correlated with missense mutations and nonsense/frameshift mutations, respectively. A predictive model for diagnosis was developed based on the correlation between TP53 mutation and p53 expression patterns. These results indicate that the scoring system based on p53 and Ki-67 expression patterns can differentiate epithelial lesions, especially in cases when the morphological features are ambiguous.

Primary cardiac sarcomas: A clinicopathologic study in a single institution with 25 years of experience with an emphasis on MDM2 expression and adjuvant therapy for prognosis.

BACKGROUND: Primary cardiac sarcomas are rare and their clinicopathologic features are heterogeneous. Among them, particularly intimal sarcoma is a diagnostic challenge due to nonspecific histologic features. Recently, MDM2 amplification reported to be a characteristic genetic event in the intimal sarcoma. In this study, we aimed to identify the types and incidence of primary cardiac sarcomas that occurred over 25 years in tertiary medical institutions, and to find clinicopatholgical significance through reclassification of diagnoses using additional immunohistochemistry (IHC). METHODS: We reviewed the primary cardiac sarcoma cases between January 1993 and June 2018 at Asan Medical Center, South Korea, with their clinicopathologic findings, and reclassified the subtypes, especially using IHC for MDM2 and then, analyzed the significance of prognosis. RESULTS: Forty-eight (6.8%) cases of a primary cardiac sarcoma were retrieved. The tumors most frequently involved the right atrium (n = 25, 52.1%), and the most frequent tumor subtype was angiosarcoma (n = 23, 47.9%). Seven cases (53.8%) were newly reclassified as an intimal sarcoma by IHC for MDM2. Twenty-nine (60.4%) patients died of disease (mean, 19.8 months). Four patients underwent a heart transplantation and had a median survival of 26.8 months. This transplantation group tended to show good clinical outcomes in the earlier stages, but this was not statistically significant (p = 0.318). MDM2 positive intimal sarcoma showed the better overall survival (p = 0.003) than undifferentiated pleomorphic sarcoma. Adjuvant treatment is beneficial for patient survival (p < 0.001), particularly in angiosarcoma (p < 0.001), but not in intimal sarcoma (p = 0.154). CONCLUSION: Our study supports the use of adjuvant treatment in primary cardiac sarcoma, as it was associated with a significantly better overall survival rate. Further consideration of tumor histology may be important in determining the optimal use of adjuvant treatment for different types of sarcomas. Therefore, accurate diagnosis by MDM2 test is important condsidering patient's prognosis and treatment.

Characterization of a Potential Probiotic Lactiplantibacillus plantarum LRCC5310 by Comparative Genomic Analysis and its Vitamin B6 Production Ability.

Safety assessment and functional analysis of probiotic candidates are important for their industrial applications. Lactiplantibacillus plantarum is one of the most widely recognized probiotic strains. In this study we aimed to determine the functional genes of L. plantarum LRCC5310, isolated from kimchi, using next-generation, whole-genome sequencing analysis. Genes were annotated using the Rapid Annotations using Subsystems Technology (RAST) server and the National Center for Biotechnology Information (NCBI) pipelines to establish the strain's probiotic potential. Phylogenetic analysis of L. plantarum LRCC5310 and related strains showed that LRCC5310 belonged to L. plantarum. However, comparative analysis revealed genetic differences between L. plantarum strains. Carbon metabolic pathway analysis based on the Kyoto Encyclopedia of Genes and Genomes database showed that L. plantarum LRCC5310 is a homofermentative bacterium. Furthermore, gene annotation results indicated that the L. plantarum LRCC5310 genome encodes an almost complete vitamin B6 biosynthetic pathway. Among five L. plantarum strains, including L. plantarum ATCC 14917T, L. plantarum LRCC5310 detected the highest concentration of pyridoxal 5'-phosphate with 88.08 ± 0.67 nM in MRS broth. These results indicated that L. plantarum LRCC5310 could be used as a functional probiotic for vitamin B6 supplementation.

Development and validation of an analytical method using liquid chromatography-tandem mass spectrometry for the therapeutic drug monitoring of seven cardiovascular drugs in clinical usage.

Therapeutic drug monitoring (TDM) of cardiovascular drugs is essential to improve treatment efficacy and minimize toxicity because of the usage of multiple drugs with a very limited therapeutic range and the high pharmacokinetic variation in patients. We developed and validated a reliable and economical liquid chromatography/tandem mass spectrometry (LC-MS/MS) method for the determination of seven cardiovascular drugs-procainamide, lidocaine, quinidine, deslanoside, digoxin, atorvastatin, and digitoxin-for clinical usage. Serum samples were prepared by simple protein precipitation with an organic solvent consisting of acetonitrile and methanol (2:1 v/v) and analyzed under optimized LC-MS/MS conditions. The chromatographic separations were accomplished within 15 min on a reversed-phase C18 column with a gradient elution of aqueous solvent and acetonitrile while maintaining 0.1 (v/v) % formic acid and 2 mM ammonium formate. The optimized MS/MS conditions in ESI-positive mode offered sufficient sensitivity for the seven cardiovascular drugs (LOQs between 0.5 and 1 ng/mL). This method was fully validated including linearity, selectivity, accuracy, precision, carry-over, and matrix effects. Additionally, stability under several conditions was tested to determine how to handle the standard solutions and serum samples. The seven cardiovascular drugs, simultaneously, were precisely and accurately analyzed in intra- and inter-day assays (RSD < 6 % and recovery between 96.3 and 102.8 %) using only two isotope-labeled internal standards (lidocaine-(diethyl-d10) and digoxin-21, 21, 22-d3). The presented method also showed good accuracy in analyzing the seven drugs in hyperlipidemia, hyperalbuminemia, and hyperglycemia serum, allowing it to be recommended as a common and routine analysis method for cardiovascular drugs in clinical practice.

Muricauda lutisoli sp. nov., isolated from mudflat.

A Gram-negative, nonmotile, rod-shaped bacterium, designated strain CAU 1631T, was isolated from a mudflat sample in the Republic of Korea. Strain CAU 1631T grew optimally at 30 °C, pH 6.5, and 1% (w/v) NaCl solution. Phylogenetic analysis based on 16S rRNA gene sequencing and 92 core genes indicated that strain CAU 1631T is a member of the genus Muricauda and most closely related to Muricauda oceanensis 40DY170T and Muricauda lutimaris SMK-108T (98.1%, both). The draft genome was 3.4 Mb with 3064 protein-coding genes, and the DNA G + C content was 43.3 mol%. The major fatty acids were iso-C15:0, iso-C17:0 3-OH, and iso-C15:0 G, and the major polar lipid was phosphatidylethanolamine. The predominant respiratory quinone was MK-6. Based on the comprehensive taxonomic characterization, strain CAU 1631T is a novel species, for which the name Muricauda lutisoli sp. nov. has been proposed. The type strain is CAU 1631T (= KCTC 82456T = MCCC 1K06088T).

Development of Certified Reference Material for Amino Acids in Dried Blood Spots and Accuracy Assessment of Disc Sampling.

To achieve the measurement reliability of amino acids used as diagnostic markers in clinical fields, establishing a reference measurement system is required, in which certified reference materials (CRMs) are an essential step in the hierarchy of measurement traceability. This study describes the development of dried blood spot (DBS) CRMs for amino acid analysis with complete measurement traceability to the International System of Units (SI). Six essential amino acids─proline, valine, isoleucine, leucine, phenylalanine, and tyrosine─were analyzed using isotope-dilution liquid chromatography-mass spectrometry (ID-MS). For minimizing measurement bias and uncertainty overestimation, whole spots with 50 µL of whole blood were adopted in the certification. The between-spot homogeneities by whole spot sampling were lower than 2.1%. The relative expanded uncertainties of the six amino acids in the developed DBS CRMs were lower than 5.7% at 95% confidence. The certified values are traceable to SI through both gravimetric preparation and the primary method in certification, ID-MS. Comparison among DBS testing laboratories revealed discrepancies between the whole spot and disc sampling methods. The actual sampling volume was accurately estimated by weighing, which revealed the possibility of underestimation in routine DBS testing. The candidate CRMs can support the standardization of DBS testing for amino acids through the qualification and validation of many kinds of measurement procedures to compensate the measurement bias caused by matrix-specific sampling error.

Halomarinibacterium sedimenti gen. nov., sp. nov., a carotenoid pigment-producing bacterium isolated from marine sediment.

A Gram-negative, strictly aerobic, non-motile, rod-shaped bacterial strain CAU 1614T was isolated from a marine sediment sample collected in the Republic of Korea. Optimal growth of strain CAU 1614T proceeded at 30 °C, pH 7.0, and 2% (w/v) NaCl. 16S rRNA gene similarity was lower than 94.5% with genera Aureisphaera, Marinirhabdus, Aureitalea, Gilvibacter, Ulvibacter, and Jejudonia. The highest similarity was with Aureisphaera galaxeae 04OKA003-7T (94.5%). The major cellular fatty acids were iso-C15:0, iso-C16:0, iso-C15:1 G, iso-C16:0 3-OH, and iso-C17:0 3-OH and the predominant menaquinone was MK-6. The polar lipids were phosphatidylethanolamine, phosphoglycolipid, an unidentified lipid, two unidentified aminolipids, and an unidentified glycolipid. The draft genome of strain CAU 1614T was 3.9 Mb and DNA G+C content was 36.0 mol%. On the basis of the phenotypic, chemotaxonomic, and genomic data, strain CAU 1614T presents a novel genus in the family Flavobacteriaceae, for which the name Halomarinibacterium sedimenti gen. nov., sp. nov. is proposed. The type strain is CAU 1614T (= KCTC 82457T = MCCC 1K06083T).

Measurement comparability of insulin assays using conventional immunoassay kits.

BACKGROUND: The standardization of measurement aims to achieve comparability of results regardless of the analytical methods and the laboratory where analyses are carried out. In this paper, a comparison of results from several immunoassay-based insulin analysis kits is described, and the steps necessary to improve comparability are discussed. METHODS: Four manual enzyme-linked immunosorbent assay (ELISA) kits produced by Mercodia, Alpco, Epitope Diagnostics, and Abcam, and three automated chemiluminescent (CLIA) insulin assay kits (Siemens Centaur XP, Unicel Dxl800, Cobas e801) were compared by analyzing human serum samples and certified reference materials for human insulin. RESULTS: The seven evaluated assay kits showed substantial discrepancies in the results, with relative standard deviation ranges between 1.7% and 23.2%. We find that the traceability chains and the unit conversion factors are not yet harmonized, and current reference materials for insulin are not applicable for immunoassay-based method validation due to the use of different matrices. CONCLUSIONS: The findings suggest the need to fine tune insulin analysis methods, measurement traceability, and any conversion factor used in post-analysis steps in accordance with the necessity for standardization.

Arenibacterium arenosum sp. nov., isolated from sea sand.

A Gram-negative, non-motile, short rod-shaped aerobic bacterial strain CAU 1593T was isolated from a coastal sand sample collected in the Republic of Korea. Cells of strain CAU 1593T grew optimally at 30 °C and pH 7.5 in 4% (w/v) NaCl. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain CAU 1593T had the highest similarity to Arenibacterium halophilum (97.5%). The whole genome of strain CAU 1593T was 3,979,826 bp with 26 contigs, and the DNA G + C content was 64.3 mol%. The major fatty acid of strain CAU 1593 T was summed feature 8 (C18:1 ω7c/C18:1 ω6c). Q-10 was the only respiratory quinone. The polar lipids were composed of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, two phosphoglycolipids, an unidentified glycolipid, and an unidentified lipid. Based on the results of chemotaxonomic, phylogenetic, and phenotypic analyses, strain CAU 1593T represents a novel species in the genus Arenibacterium, which is named Arenibacterium arenosum sp. nov. The type strain is CAU 1593T (= KCTC 82402T = MCCC 1K05671T).

Lactiplantibacillus plantarum LRCC5314 includes a gene for serotonin biosynthesis via the tryptophan metabolic pathway.

As the functions of probiotics within the same species may not be shared, it is important to analyze the genetic characteristics of strains to determine their safety and usefulness before industrial applications. Hence the present study was undertaken to determine functional genes, and beneficial activities of strain LRCC5314, a bacterial strain isolated from kimchi through comparative genomic analysis. Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain LRCC5314 was a member of the species L. plantarum. Whole genome size of strain LRCC5314 was sequence was 3.25 Mb long, with a G + C content of 44.5 mol% and 3,031 predicted genes. Strain LRCC5314 could metabolize hexoses through homofermentation, which produces only lactic acid from hexoses. According to gene annotation, strain LRCC-5314 contained genes of EPS production and CRISPR. Moreover, the strain contained genes that could encode a complete biosynthetic pathway for the production of tryptophan, which can be used as a precursor of serotonin. Notably, the tryptophan and serotonin activities strain LRCC5314 were higher than those of reference strains, L. plantarum ATCC 14917T, DSM 20246, DSM 2601, and ATCC 8014, which reach tryptophan amount of 0.784 ± 0.045 µM/ml in MRS broth and serotonin concentration of 19.075 ± 0.295 ng/ml in HT-22 cells. These findings indicated that L. plantarum LRCC5314 could provide a source for serotonin production and could be used as a functional probiotic for stress regulation.

Clinico-cytopathologic analysis of 574 Pericardial Effusion Specimens: Application of the international system for reporting serous fluid cytopathology (ISRSFC) and long-term clinical follow-up.

INTRODUCTION: A pericardial effusion (PE) has a variable etiology and the primary role is diagnosis of metastatic malignancy. We analyzed the PE cytology in a large cohort in accordance with the international system for reporting serous fluid cytopathology (ISRSFC) and evaluated the long-term patient outcomes. METHODS: PE specimens from 2010 to 2014 with an available clinical history, cytologic data, and pericardial biopsy results were collected. RESULTS: A total of 574 PE specimens were obtained from 486 patients, representing 1.5% (574/38,589) of all body fluid specimens. Three hundred and eighty-two (66.6%) cases were "negative," 54 (9.4%) cases were "atypia of undetermined significance," 10 (1.7%) cases were "suspicious for malignancy," and 128 (22.3%) cases were "malignancy". The most common origin for malignant PE was the lung (82.1%), in both men (70.5%) and women (50.6%). Breast cancer (20%) in women and gastric cancer (4.9%) in men were the second most common malignant PE, respectively. The mean interval from the occurrence of malignant PE to death was 10.06 months (range; 0-116.03 months, median 3.5 months), and the 1-year survival rate was 16.7%. In addition, the 1-year survival rates after malignant PE onset were 0% for gastric cancer, 13.9% for lung cancer, 19.8% for breast cancer, and 21.1% for the other cancers (p = 0.011). CONCLUSION: Our present study is the first to our knowledge to classify the pericardial fluid from 574 cases in accordance with the recently published ISRSFC, and to present the long-term outcomes of patients with malignant PE at the same time. Moreover, we report for the first time that it is gastric and not lung cancer patients that have the poorest prognosis after the occurrence of malignant PE.