Characterization of 3D Organotypic Culture of Mouse Adipose-Derived Stem Cells.

Although stem cells are a promising avenue for harnessing the potential of adipose tissue, conventional two-dimensional (2D) culture methods have limitations. This study explored the use of three-dimensional (3D) cultures to preserve the regenerative potential of adipose-derived stem cells (ADSCs) and investigated their cellular properties. Flow cytometric analysis revealed significant variations in surface marker expressions between the two culture conditions. While 2D cultures showed robust surface marker expressions, 3D cultures exhibited reduced levels of CD44, CD90.2, and CD105. Adipogenic differentiation in 3D organotypic ADSCs faced challenges, with decreased organoid size and limited activation of adipogenesis-related genes. Key adipocyte markers, such as lipoprotein lipase (LPL) and adipoQ, were undetectable in 3D-cultured ADSCs, unlike positive controls in 2D-cultured mesenchymal stem cells (MSCs). Surprisingly, 3D-cultured ADSCs underwent mesenchymal-epithelial transition (MET), evidenced by increased E-cadherin and EpCAM expression and decreased mesenchymal markers. This study highlights successful ADSC organoid formation, notable MSC phenotype changes in 3D culture, adipogenic differentiation challenges, and a distinctive shift toward an epithelial-like state. These findings offer insights into the potential applications of 3D-cultured ADSCs in regenerative medicine, emphasizing the need for further exploration of underlying molecular mechanisms.

Restless legs syndrome in patients with obstructive sleep apnea: Association between apnea severity and symptoms of depression, insomnia, and daytime sleepiness.

OBJECTIVE: To determine if the prevalence and severity of restless legs syndrome (RLS) varies with apnea severity and analyze differences between the sexes in terms of comorbid RLS with symptoms of depression, insomnia, and daytime sleepiness in patients with obstructive sleep apnea (OSA). METHODS: Symptoms of depression, insomnia, and daytime sleepiness were defined as Patient Health Questionnaire-9 score ≥10, Insomnia Severity Index score ≥15, and Epworth Sleepiness Scale score ≥11. Multivariate logistic and linear regression analyses were conducted. RESULTS: In 707 adults with OSA (85.1% males), 16.1% (n = 114) had comorbid RLS. The prevalence of RLS was markedly lower in those with moderate and severe OSA than in those with mild OSA. Similarly, the odds of RLS significantly decreased with increasing apnea-hypopnea index. After controlling for age and sex, in patients with comorbid RLS, the International RLS Study Group Rating Scale scores were negatively correlated with apnea-hypopnea index and a nadir peripheral oxygen saturation during sleep. The presence of RLS was more likely to be associated with symptoms of depression, insomnia, and daytime sleepiness after controlling for confounding variables, but only in men. CONCLUSIONS: RLS is frequently noted in combination with OSA, with a female preponderance. The severities of OSA and RLS may be negatively associated. In patients with OSA, sex-related differences in terms of comorbid RLS with symptoms of depression, insomnia, and daytime sleepiness warrant further investigations.

Sarcopenia with decreased total psoas muscle area in children with high-risk neuroblastoma.

BACKGROUND: We calculated psoas muscle area (PMA) z-scores in high-risk neuroblastoma patients undergoing treatment to examine the clinical significance of sarcopenia in this cohort. METHODS: We analyzed retrospective data from patients aged 0-18 who were diagnosed with abdominal neuroblastoma between 2005 and 2019 at Samsung Medical Center. Patients categorized as high-risk undergone induction chemotherapy, neuroblastoma excision, and tandem high-dose chemotherapy with autologous stem cell transplantation (HDCT/auto-SCT) were selected. L3-4 lumbar levels on axial CT images were identified and we measured the areas of the left and right psoas muscles to determine tPMA. Total PMA z-scores were calculated using an open online tool. RESULTS: There were 45 boys and 25 girls with a mean age of 3.86 years. CT images taken at initial diagnosis and after tandem HDCT/auto-SCT were selected to calculate tPMA z-scores. Mean elapsed time between the two measurements was 12.91 ± 1.73 months. Mean tPMA z-score significantly decreased from -0.21 ± 1.29 to -0.66 ± 0.97 (p = 0.022). Length of hospital stay was significantly longer in the group of patients whose tPMA z-scores decreased by more than .45 (177.62 ± 28.82 days vs. 165.75 ± 21.34 days, p = 0.049). Presence of sarcopenia at initial diagnosis was a significant risk factor for bacterial infection during neuroblastoma treatment. CONCLUSION: tPMA z-scores in high-risk neuroblastoma patients decreased significantly following a treatment regimen that included induction chemotherapy, tumor resection surgery, and HDCT/auto-SCT. A greater decrease in tPMA z-score was associated with longer hospital stay during treatment.

Multitask Learning-Based Deep Signal Identification for Advanced Spectrum Sensing.

The explosive demand for wireless communications has intensified the complexity of spectrum dynamics, particularly within unlicensed bands. To promote efficient spectrum utilization and minimize interference during communication, spectrum sensing needs to evolve to a stage capable of detecting multidimensional spectrum states. Signal identification, which identifies each device's signal source, is a potent method for deriving the spectrum usage characteristics of wireless devices. However, most existing signal identification methods mainly focus on signal classification or modulation classification, thus offering limited spectrum information. In this paper, we propose DSINet, a multitask learning-based deep signal identification network for advanced spectrum sensing systems. DSINet addresses the deep signal identification problem, which involves not only classifying signals but also deriving the spectrum usage characteristics of signals across various spectrum dimensions, including time, frequency, power, and code. Comparative analyses reveal that DSINet outperforms existing shallow signal identification models, with performance improvements of 3.3% for signal classification, 3.3% for hall detection, and 5.7% for modulation classification. In addition, DSINet solves four different tasks with a 65.5% smaller model size and 230% improved computational performance compared to single-task learning model sets, providing meaningful results in terms of practical use.

Complete genome sequence of an oryctes rhinoceros nudivirus isolated from Korean rhinoceros beetles (Trypoxylus dichotomus) in Korea.

Nudivirus-infected Korean rhinoceros beetles (Trypoxylus dichotomus) were first identified in 2015, and while a complete genome sequence of the virus has long been uploaded to the NCBI database, it has not been examined in detail. Here, we describe the genomic characteristics of Trypoxylus dichotomus nudivirus (TdNV), which represents a new Oryctes rhinoceros nudivirus (OrNV) strain, isolated from infected T. dichotomus in the Republic of Korea. We examined factors derived by the cross-species infection of OrNV from nucleotide levels to the whole genome level. Our genomic analysis study suggests that TdNV-Korea is highly conserved with other OrNVs in terms of genomic structures and genome size. Our investigation of the genomic structure revealed that TdNV-Korea has the least number of open reading frames (ORFs) of all available OrNV genomes; three hypothetical genes were notably absent only in TdNV-Korea. In addition, the genomic alteration of the nudivirus core genes discloses that various amino acid mutations caused by single-nucleotide polymorphism and short indels (insertion/deletion) were found in most of the nudivirus core genes of TdNV-Korea. Our findings provide a valuable resource for those seeking a greater understanding of cross-species nudivirus transmission and will certainly provide valuable insight for reconstruction and reinterpretation of future and previously identified OrNV strains.

Developmental delay and non-phenylketonuria (PKU) hyperphenylalaninemia in DNAJC12 deficiency: Case and approach.

BACKGROUND: Hyperphenylalaninemia is a biomarker for several monogenic neurotransmitter disorders where the body cannot metabolise phenylalanine to tyrosine. Biallelic pathogenic variants in DNAJC12, co-chaperone of phenylalanine, tyrosine, and tryptophan hydroxylases, leads to hyperphenylalaninemia and biogenic amines deficiency. METHODS AND RESULTS: A male firstborn to non-consanguineous Sudanese parents had hyperphenylalaninemia 247 µmol/L [reference interval (RI) < 200 µmol/L] at newborn screening. Dried blood spot dihydropteridine reductase (DHPR) assay and urine pterins were normal. He had severe developmental delay and autism spectrum disorder without a notable movement disorder. A low phenylalanine diet was introduced at two years without any clinical improvements. Cerebrospinal fluid (CSF) neurotransmitters at five years demonstrated low homovanillic acid (HVA) 0.259 µmol/L (reference interval (RI) 0.345-0.716) and 5-hydroxyindoleaetic acid (5HIAA) levels 0.024 µmol/L (reference interval (RI) 0.100-0.245). Targeted neurotransmitter gene panel analysis identified a homozygous c.78 + 1del variant in DNAJC12. At six years, he was commenced on 5-hydroxytryptophan 20 mg daily, and his protein-restricted diet was liberalised, with continued good control of phenylalanine levels. Sapropterin dihydrochloride 7.2 mg/kg/day was added the following year with no observable clinical benefits. He remains globally delayed with severe autistic traits. CONCLUSIONS: Urine, CSF neurotransmitter studies, and genetic testing will differentiate between phenylketonuria, tetrahydrobiopterin or DNAJC12 deficiency, with the latter characterised by a clinical spectrum ranging from mild autistic features or hyperactivity to severe intellectual disability, dystonia, and movement disorder, normal DHPR, reduced CSF HIAA and HVA. DNAJC12 deficiency should be considered early in the differential workup of hyperphenylalaninemia identified from newborn screening, with its genotyping performed once deficiencies of phenylalanine hydroxylase (PAH) and tetrahydrobiopterin (BH4) have been biochemically or genetically excluded.

Evaluation of Radiation Sensitivity Differences in Mouse Liver Tumor Organoids Using CRISPR/Cas9-Mediated Gene Mutation.

BACKGROUND: To assess the radiosensitivity of liver tumors harboring different genetic mutations, mouse liver tumors were generated in vivo through the hydrodynamic injection of clustered regularly interspaced short palindromic repeat/caspase 9 (CRISPR/Cas9) constructs encoding single-guide RNAs (sgRNAs) targeting Tp53, Pten, Nf1, Nf2, Tsc2, Cdkn2a, or Rb1. METHODS: The plasmid vectors were delivered to the liver of adult C57BL/6 mice via hydrodynamic tail vein injection. The vectors were injected into 10 mice in each group. Organoids were generated from mouse liver tumors. The radiation response of the organoids was assessed using an ATP cell viability assay. RESULTS: The mean survival period of mice injected with vectors targeting Nf2 (4.8 months) was lower than that of other mice. Hematoxylin and eosin staining, immunohistochemical (IHC) staining, and target sequencing analyses revealed that mouse liver tumors harbored the expected mutations. Tumor organoids were established from mouse liver tumors. Histological evaluation revealed marked morphological similarities between the mouse liver tumors and the generated tumor organoids. Moreover, IHC staining indicated that the parental tumor protein expression pattern was maintained in the organoids. The results of the ATP cell viability assay revealed that the tumor organoids with mutated Nf2 were more resistant to high-dose radiation than those with other gene mutations. CONCLUSIONS: This study developed a radiation response assessment system for mouse tumors with mutant target genes using CRISPR/Cas9 and organoids. The Tp53 and Pten double mutation in combination with the Nf2 mutation increased the radiation resistance of tumors. The system used in this study can aid in elucidating the mechanism underlying differential intrinsic radiation sensitivity of individual tumors.

Automatic Thinning Detection through Image Segmentation Using Equivalent Array-Type Lamp-Based Lock-in Thermography.

Among the non-destructive testing (NDT) techniques, infrared thermography (IRT) is an attractive and highly reliable technology that can measure the thermal response of a wide area in real-time. In this study, thinning defects in S275 specimens were detected using lock-in thermography (LIT). After acquiring phase and amplitude images using four-point signal processing, the optimal excitation frequency was calculated. After segmentation was performed on each defect area, binarization was performed using the Otsu algorithm. For automated detection, the boundary tracking algorithm was used. The number of pixels was calculated and the detectability using RMSE was evaluated. Clarification of defective objects using image segmentation detectability evaluation technique using RMSE was presented.

DRL-OS: A Deep Reinforcement Learning-Based Offloading Scheduler in Mobile Edge Computing.

Hardware bottlenecks can throttle smart device (SD) performance when executing computation-intensive and delay-sensitive applications. Hence, task offloading can be used to transfer computation-intensive tasks to an external server or processor in Mobile Edge Computing. However, in this approach, the offloaded task can be useless when a process is significantly delayed or a deadline has expired. Due to the uncertain task processing via offloading, it is challenging for each SD to determine its offloading decision (whether to local or remote and drop). This study proposes a deep-reinforcement-learning-based offloading scheduler (DRL-OS) that considers the energy balance in selecting the method for performing a task, such as local computing, offloading, or dropping. The proposed DRL-OS is based on the double dueling deep Q-network (D3QN) and selects an appropriate action by learning the task size, deadline, queue, and residual battery charge. The average battery level, drop rate, and average latency of the DRL-OS were measured in simulations to analyze the scheduler performance. The DRL-OS exhibits a lower average battery level (up to 54%) and lower drop rate (up to 42.5%) than existing schemes. The scheduler also achieves a lower average latency of 0.01 to >0.25 s, despite subtle case-wise differences in the average latency.

3-Methylglutaconyl-CoA hydratase deficiency: When ascertainment bias confounds a biochemical diagnosis.

3-Methylglutaconyl-CoA hydratase deficiency (MGA1) is a defect in leucine catabolism, which causes the accumulation of urinary 3-methylglutaconate, with or without 3-hydroxyisovalerate and 3-methylglutarate. It is an ultra-rare condition, with <30 cases published in the literature. It is unclear whether the clinical features seen in reported patients are caused by the biochemical abnormalities, or whether they simply represent an ascertainment bias in patients that come to clinical attention. We reviewed the collective Australian experience of patients with confirmed MGA1, four of whom were diagnosed when asymptomatic through newborn screening (NBS). When our cohort is considered alongside the broader literature, there is no clear evidence of a specific childhood-onset clinical phenotype associated with this disorder. Some patients have non-specific clinical features (such as autism spectrum disorder [ASD]); however, there are also other family members with ASD in the absence of MGA1, suggesting a multifactorial aetiology. Importantly, all four patients diagnosed through NBS (including three with over 18 years of clinical follow-up) remain asymptomatic in the absence of treatment. Based on the available literature, we suggest that MGA1 represents a biochemical phenotype, with an absence of a childhood clinical phenotype. The burdens of sustained treatment (particularly with intensive dietary leucine restriction) in asymptomatic individuals may be of little benefit, and likely to result in poor compliance. Longer-term follow-up of patients detected via NBS (or biochemical screening of large cohorts of asymptomatic adult individuals) will be required to conclusively prove or disprove the association with adult-onset leukoencephalopathy.

Secretory SERPINE1 Expression Is Increased by Antiplatelet Therapy, Inducing MMP1 Expression and Increasing Colon Cancer Metastasis.

Contrary to many reports that antiplatelet agents inhibit cancer growth and metastasis, new solid tumors have been reported in patients receiving long-term antiplatelet therapy. We investigated the effects of these agents directly on cancer cells in the absence of platelets to mimic the effects of long-term therapy. When four antiplatelet agents (aspirin, clopidogrel, prasugrel, and ticagrelor) were administered to colon cancer cells, cancer cell proliferation was inhibited similarly to a previous study. However, surprisingly, when cells were treated with a purinergic P2Y12 inhibitor (purinergic antiplatelet agent), the motility of the cancer cells was significantly increased. Therefore, gene expression profiles were identified to investigate the effect of P2Y12 inhibitors on cell mobility, and Serpin family 1 (SERPINE1) was identified as a common gene associated with cell migration and cell death in three groups. Antiplatelet treatment increased the level of SERPINE1 in cancer cells and also promoted the secretion of SERPINE1 into the medium. Increased SERPINE1 was found to induce MMP1 and, thus, increase cell motility. In addition, an increase in SERPINE1 was confirmed using the serum of patients who received these antiplatelet drugs. With these results, we propose that SERPINE1 could be used as a new target gene to prevent the onset and metastasis of cancer in patients with long-term antiplatelet therapy.

A Semantic Data-Based Distributed Computing Framework to Accelerate Digital Twin Services for Large-Scale Disasters.

As natural disasters become extensive, due to various environmental problems, such as the global warming, it is difficult for the disaster management systems to rapidly provide disaster prediction services, due to complex natural phenomena. Digital twins can effectively provide the services using high-fidelity disaster models and real-time observational data with distributed computing schemes. However, the previous schemes take little account of the correlations between environmental data of disasters, such as landscapes and weather. This causes inaccurate computing load predictions resulting in unbalanced load partitioning, which increases the prediction service times of the disaster management agencies. In this paper, we propose a novel distributed computing framework to accelerate the prediction services through semantic analyses of correlations between the environmental data. The framework combines the data into disaster semantic data to represent the initial disaster states, such as the sizes of wildfire burn scars and fuel models. With the semantic data, the framework predicts computing loads using the convolutional neural network-based algorithm, partitions the simulation model into balanced sub-models, and allocates the sub-models into distributed computing nodes. As a result, the proposal shows up to 38.5% of the prediction time decreases, compared to the previous schemes.

A Novel Digital Twin Architecture with Similarity-Based Hybrid Modeling for Supporting Dependable Disaster Management Systems.

Disaster management systems require accurate disaster monitoring and prediction services to reduce damages caused by natural disasters. Digital twins of natural environments can provide the services for the systems with physics-based and data-driven disaster models. However, the digital twins might generate erroneous disaster prediction due to the impracticability of defining high-fidelity physics-based models for complex natural disaster behavior and the dependency of data-driven models on the training dataset. This causes disaster management systems to inappropriately use disaster response resources, including medical personnel, rescue equipment and relief supplies, to ensure that it may increase the damages from the natural disasters. This study proposes a digital twin architecture to provide accurate disaster prediction services with a similarity-based hybrid modeling scheme. The hybrid modeling scheme creates a hybrid disaster model that compensates for the errors of physics-based prediction results with a data-driven error correction model to enhance the prediction accuracy. The similarity-based hybrid modeling scheme reduces errors from the data dependency of the hybrid model by constructing a training dataset using similarity assessments between the target disaster and the historical disasters. Evaluations in wildfire scenarios show that the digital twin decreases prediction errors by approximately 50% compared with those of the existing schemes.

Binarization Mechanism Evaluation for Water Ingress Detectability in Honeycomb Sandwich Structure Using Lock-In Thermography.

The growing use of composite honeycomb structures in several industries including aircraft has demonstrated the need to develop effective and efficient non-destructive evaluation methods. In recent years, active thermography has attracted great interest as a reliable technology for non-destructive testing and evaluation of composite materials due to its advantages of non-contact, non-destructive, full-area coverage, high speed, qualitative, and quantitative testing. However, non-uniform heating, low spatial resolution, and ambient environmental noise make the detection and characterization of defects challenging. Therefore, in this study, lock-in thermography (LIT) was used to detect water ingress into an aircraft composite honeycomb sandwich structure, and the phase signals were binarized through the Otsu algorithm. A square composite honeycomb with dimensions of 210 mm × 210 mm along with 16 different defective areas of various sizes in groups filled with water by 25%, 50%, 75%, and 100% of the cell volume was considered. The sample was excited at multiple modulation frequencies (i.e., 1 Hz to 0.01 Hz). The results were compared in terms of phase contrast and CNR according to the modulation frequency. In addition, the detectability was analyzed by comparing the number of pixels of water ingress in the binarized image and the theoretical calculation.

Hydrogen-Bonding-Mediated Molecular Vibrational Suppression for Enhancing the Fluorescence Quantum Yield Applicable for Visual Phenol Detection.

It is generally accepted that while efficient suppression of molecular vibration is inevitable for purely organic phosphors due to their long emission lifetime in the regime of 1 ms or longer, fluorophores having a lifetime in the nanoseconds regime are not sensitive to collisional quenching. Here, however, we demonstrate that a fluorophore, 2,5-bis(hexyloxy)terephthaldehyde (BHTA), capable of having hydrogen bonding (H bonding) via its two aldehyde groups can have a largely enhanced (450%) fluorescence quantum yield (QY) in amorphous poly(acrylic acid) (PAA) matrix compared to its crystalline powder. We ascribe this enhanced QY to the efficient suppression of molecular vibrations via intermolecular H bonding. We confirm this feasibility by conducting temperature-dependent fluorescence emission intensity measurement. As gaseous phenol can intervene with the H bonding between BHTA and PAA, interestingly, BHTA embedded in PAA can selectively detect gaseous phenol by a sharp fluorescence emission intensity drop that is visibly recognizable by the naked eye. The results provide an insightful molecular design strategy for a fluorophore and fluorometric sensory system design for enhanced photoluminescence QY and convenient detection of various volatile organic compounds.

Dynamic Oxygen Conditions Promote the Translocation of HIF-1α to the Nucleus in Mouse Blastocysts.

Oxygen tension is one of the most critical factors for mammalian embryo development and its survival. The HIF protein is an essential transcription factor that activated under hypoxic conditions. In this study, we evaluated the effect of dynamic oxygen conditions on the expression of embryonic genes and translocation of hypoxia-inducible factor-1α (HIF-1α) in cultured mouse blastocysts. Two-pronuclear (2PN) zygotes harvested from ICR mice were subjected to either high oxygen (HO; 20%), low oxygen (LO; 5%), or dynamic oxygen (DO; 5% to 2%) conditions. In the DO group, PN zygotes were cultured in 5% O2 from days 1 to 3 and then in 2% O2 till day 5 after hCG injection. On day 5, the percentage of blastocysts in the cultured embryos from each group was estimated, and the embryos were also subjected to immunocytochemical and gene expression analysis. We found that the percentage of blastocysts was similar among the experimental groups; however, the percentage of hatching blastocysts in the DO and LO groups was significantly higher than that in the HO group. The total cell number of blastocysts in the DO group was significantly higher than that of both the HO and LO groups. Further, gene expression analysis revealed that the expression of genes related to the embryonic development was significantly higher in the DO group than that in the HO and LO groups. Interestingly, HIF-1α mRNA expression did not significantly differ; however, HIF-1α protein translocation from the cytoplasm to the nucleus was significantly higher in the DO group than in the HO and LO groups. Our study suggests that dynamic oxygen concentrations increase the developmental capacity in mouse preimplantation embryos through activation of the potent transcription factor HIF-1α.

Prediction of Recurrence of Completely Resected Pancreatic Solid Pseudopapillary Neoplasms in Pediatric Patients: A Single Center Analysis.

BACKGROUND: Many previous studies have investigated the risk factors for the recurrence of pancreatic solid pseudopapillary neoplasms (SPNs), although a consensus has not yet been reached, despite this effort. We aimed to identify the predictive factors for recurrence in patients with SPNs who underwent complete surgical resection of the tumor. METHODS: We retrospectively analyzed the records of pediatric patients with SPNs who underwent surgical resection at a single center between 2001 and 2018. RESULTS: During the study period, 47 patients with SPNs underwent radical resection of the tumor. The median age of the patients was 14 (8-18) years. R0 resection was confirmed in every case and none of the patients presented with systemic metastasis at the time of diagnosis. The median follow-up period was 53.1 (30.8-150.8) months. Of the 47 patients, only two (4.2%) experienced recurrence. Using comparative analysis, we found that some factors such as a large tumor size, peripancreatic tissue invasion, and capsule invasion did not increase the risk of recurrence of SPNs. Lymph node metastasis was the only significant factor for recurrence in our study (p = 0.043). CONCLUSION: During our single center analysis, we found that only lymph node metastasis was a predictive factor for recurrence of SPNs among patients who underwent complete tumor resection. Long-term follow-up is required to determine whether SPNs will recur if lymph node metastasis is observed after surgery. Furthermore, therapeutic benefits of routine lymphadenectomy or sentinel lymph node biopsy should be investigated in future studies to reduce the risk of recurrence in patients with SPNs.

Abnormal Orientation of the Superior Mesenteric Vessels Detected in Asymptomatic Infants: What Is Its Destiny?

Background: Ultrasonography (USG) has been described as an alternative diagnostic tool for malrotation that evaluates the orientation of the superior mesenteric vessels. However, literature concerning the management of patients who do not have abdominal symptoms is limited. We aimed to review the clinical course of infants showing abnormal orientation of the superior mesenteric vessels on USG who were asymptomatic at the time of diagnosis. Methods: Seventy asymptomatic infants with abnormal orientation of the superior mesenteric vessels in a single center between 2014 and 2018 were retrospectively analyzed. Results: The 70 patients, 21 underwent upper gastrointestinal series (UGIS) and 11 underwent abdominal surgery for other surgical conditions. Among the 32 (45.7%) patients who underwent UGIS or abdominal surgery, 11 were proven to have malrotation. Of the 38 (54.3%) patients who did not undergo UGIS or abdominal surgery, six patients were too unstable to undergo UGIS, five died due to cardiac complications, and the remaining patient developed midgut volvulus and died 3 days after emergency surgery. The remaining 32 patients who did not undergo UGIS or abdominal surgery were discharged without additional tests, and all were asymptomatic until their last follow-up. In the multivariate analysis, history of heart surgery and the presence of more than three anomalies were significantly associated with malrotation. Conclusion: A significant number of malrotation were diagnosed in asymptomatic infants with abnormal orientation of the superior mesenteric vessels on USG. Infants with major cardiac or multiple anomalies need special attention and should undergo UGIS in a promptly manner to confirm malrotation.

Detrimental effects of lipopolysaccharide on the attachment and outgrowth of various trophoblastic spheroids on human endometrial epithelial cells.

OBJECTIVE: Lipopolysaccharide (LPS) from Gram-negative bacteria causes poor uterine receptivity by inducing excessive inflammation at the maternal-fetal interface. This study aimed to investigate the detrimental effects of LPS on the attachment and outgrowth of various types of trophoblastic spheroids on endometrial epithelial cells (ECC-1 cells) in an in vitro model of implantation. METHODS: Three types of spheroids with JAr, JEG-3, and JAr mixed JEG-3 (JmJ) cells were used to evaluate the effect of LPS on early implantation events. ECC-1 cells were treated with LPS to mimic endometrial infection, and the expression of inflammatory cytokines and adhesion molecules was analyzed by quantitative real-time polymerase chain reaction and western blotting. The attachment rates and outgrowth areas were evaluated in the various trophoblastic spheroids and ECC-1 cells treated with LPS. RESULTS: LPS treatment significantly increased the mRNA expression of inflammatory cytokines (CXCL1, IL-8, and IL-33) and decreased the protein expression of adhesion molecules (ITGß3 and ITGß5) in ECC-1 cells. The attachment rates of JAr and JmJ spheroids on ECC-1 cells significantly decreased after treating the ECC-1 cells with 1 and 10 µg/mL LPS. In the outgrowth assay, JAr spheroids did not show any outgrowth areas. However, the outgrowth areas of JEG-3 spheroids were similar regardless of LPS treatment. LPS treatment of JmJ spheroids significantly decreased the outgrowth area after 72 hours of coincubation. CONCLUSION: An in vitro implantation model using novel JmJ spheroids was established, and the inhibitory effects of LPS on ECC-1 endometrial epithelial cells were confirmed in the early implantation process.