Hemophagocytic lymphohistiocytosis associated with acute otitis media: A case report.

INTRODUCTION: Hemophagocytic lymphohistiocytosis (HLH) is a potentially life-threatening syndrome for which early recognition and treatment are essential for improving outcomes. HLH is characterized by uncontrolled immune activation leading to fever, cytopenias, hepatosplenomegaly, coagulation abnormalities, and elevated typical markers. This condition can be genetic or secondary, with the latter often triggered by infections. Here, we present a unique case of HLH secondary to acute otitis media (AOM), a common ear infection. PATIENT CONCERNS: We describe a 4-year-old boy who initially presented with a high fever and otalgia, later diagnosed with bilateral AOM. Despite antibiotic treatment, his condition deteriorated. DIAGNOSIS: The patient fulfilled diagnostic criteria for HLH. INTERVENTIONS: Aggressive treatment by using combination therapy with immunoglobulins, intravenous steroids (dexamethasone), cyclosporine, and etoposide was performed. OUTCOMES: After 1 month of treatment, improvement in the otologic symptoms was observed, and hematological findings gradually improved and normalized. LESSIONS: The link between AOM and HLH may be associated with inflammatory responses and immunological mechanisms, highlighting the importance of considering HLH in severe infection cases. This case emphasizes the need for prompt diagnosis and management, especially in secondary HLH scenarios, to improve patient outcomes. It is imperative to be aware of the potential correlation between these 2 conditions, and healthcare professionals should consider the likelihood of HLH.

Generation of sarconoids from angiosarcoma patients as a systematic-based rational approach to treatment.

Angiosarcoma is a rare subtype of malignant neoplasm originating from vascular or lymphatic endothelial cells; its low incidence has posed significant challenges for comprehensive investigations into its pathogenic mechanisms and the development of innovative treatment modalities through in vitro and in vivo models. Recent endeavors spearheaded by patient-partnered research initiatives have aimed to elucidate the intricacies of angiosarcomas by leveraging biological omics approaches, with the overarching objective of enhancing prognostic indicators and therapeutic options for this uncommon pathology. To bridge the gap between preclinical research and translational applications, we engineered angiosarcoma-derived organoids from surgically resected primary tumors, hereafter referred to as "sarconoids," as a proof-of-concept model. A novel protocol for the establishment of these sarconoids has been developed and validated. To ensure that the sarconoids faithfully recapitulate the heterogeneity and complexities of the patients' original tumors, including transcriptomic signatures, cell-type specificity, and morphological traits, exhaustive histological and transcriptomic analyses were conducted. Subsequently, we expanded the scope of our study to include an evaluation of a sarconoid-based drug screening platform; for this purpose, a drug library (AOD IX), supplied by the National Cancer Institute's Developmental Therapeutics Program, was screened using 96-well plates. Our findings suggest that sarconoids can be reliably generated from angiosarcoma patient-derived tissues and can serve as accurate models for evaluating therapeutic responses, thereby holding far-reaching implications for translational research and clinical applications aimed at advancing our understanding and treatment of angiosarcoma.

Therapeutic effect of intraperitoneal dexamethasone on noise-induced permanent threshold shift in mice model.

The purpose of the study was to which investigate whether dexamethasone, which has anti-inflammatory and immune response suppression roles, could treat noise-induced hearing loss caused by damage to hair cells in the cochlea. The experiment used 8-week-old CBA mice exposed to white noise at an intensity of 110 dB SPL for 2 h, with hearing loss confirmed by the auditory brainstem response test. Dexamethasone was administered by intraperitoneal injection for 5 days, and the therapeutic effect was investigated for 3 weeks. The experimental groups were 3 mg/kg of dexamethasone (3 mpk) and 10 mg/kg of dexamethasone (10 mpk), and the control group was a saline-administered group. The results showed that compared to the control group, the hearing threshold value was recovered by 10 dB SPL compared to the saline group from the 14th day in the 3 mpk group. In the 10 mpk group, thresholds were recovered from the 7th day compared to the saline group. This difference was similar at 4 kHz, and in the case of the 10 mpk group, the threshold was recovered by 20 dB SPL compared to the saline group. The study also confirmed the restoration of nerve cell activity and showed a recovery effect of about 20 µV in the amplitude value change in the 10 mpk group. In conclusion, the study suggests that dexamethasone has a therapeutic effect for noise-induced hearing loss by increasing the activity of nerve cells and showing a recovery effect from hair cells damaged by noise.

Recent Updates on Tinnitus Management.

In this comprehensive review, we discuss recent updates on tinnitus evaluation and treatment. Tinnitus evaluation commences with comprehensive medical history taking and audiological evaluation, which can provide valuable insight into the nature and extent of auditory disturbances. Additionally, tinnitus evaluation includes investigation of psychosomatic comorbidities to determine the intricate interplay between psychological factors and tinnitus perception. Various therapeutic approaches are available to minimize the burden of tinnitus. Cognitive behavioral therapy reshapes negative thought patterns and behaviors that are closely associated with tinnitus-induced distress. Acceptance and commitment therapy fosters mindfulness and value-aligned actions to address emotional effects. Tinnitus retraining therapy combines counseling and sound therapy for habituation. Tailor-made notched music therapy offers customized auditory experiences for symptom relief. Hearing aids and cochlear implants compensate for hearing loss and associated stress. Both neuromodulation and neurofeedback may be potentially useful. The role of pharmacotherapy and dietary supplements remains uncertain. Physiotherapy and head-neck manipulation relieve tinnitus associated with orofacial factors. Virtual reality, smartphone applications, and photobiomodulation may serve as novel therapeutic avenues. Although promising interventions are available, further research is warranted to confirm their effectiveness and long-term effects.

Cochlear Implantation in the Elderly: Speech Performance, Associated Factor, Complication, and Surgical Safety.

BACKGROUND AND OBJECTIVES: The guidelines for cochlear implantation (CIs) are expanding, and the number of CI procedures performed on the elderly is increasing. The purpose of this study was to analyze the results and safety of cochlear implantation in the elderly, as well as to evaluate the predictive factors on CI outcomes. SUBJECTS AND METHODS: The study included 56 patients aged ≥40 years, who received CIs between 2009 and 2020. They were divided into two groups: 27 younger adults (40-64 years) and 29 elderly (>64 years). The study compared their pre- and postoperative speech perception and category of auditory performance (CAP) scores, surgical complications, and hospitalization periods. It also evaluated associated factors in the elderly group by examining categorical and continuous variables and postoperative CAP score. RESULTS: There was a significant improvement in speech recognition tests (both word and sentence) and CAP scores in both groups compared to the pre-implantation scores (p<0.001). Postoperative results were slightly lower in the elderly group than in younger adults for sentence recognition and CAP scores, except for word recognition. No significant associated factors were found on postoperative CAP scores, except for etiology. Postoperative CAP significantly improved in the sudden hearing loss group compared to the groups with other etiologies (p=0.045). The elderly group had more comorbidities than that in the younger adult group (p=0.026), but there were no significant differences in postoperative complications and hospitalization periods. CONCLUSIONS: While speech recognition and CAP scores were relatively lower in the elderly group compared to the younger adults, the elderly group showed significant improvements in audiological results after CI. Moreover, CI was safe and well tolerated in elderly patients.

Association between fatty liver disease and hearing impairment in Korean adults: a retrospective cross-sectional study.

BACKGROUND: We hypothesized that fatty liver disease (FLD) is associated with a high prevalence of hearing loss (HL) owing to metabolic disturbances. This study aimed to evaluate the association between FLD and HL in a large sample of the Korean population. METHODS: We used a dataset of adults who underwent routine voluntary health checkups (n=21,316). Fatty liver index (FLI) was calculated using Bedogni's equation. The patients were divided into two groups: the non-FLD (NFLD) group (n=18,518, FLI <60) and the FLD group (n=2,798, FLI ≥60). Hearing thresholds were measured using an automatic audiometer. The average hearing threshold (AHT) was calculated as the pure-tone average at four frequencies (0.5, 1, 2, and 3 kHz). HL was defined as an AHT of >40 dB. RESULTS: HL was observed in 1,370 (7.4%) and 238 patients (8.5%) in the NFLD and FLD groups, respectively (p=0.041). Compared with the NFLD group, the odds ratio for HL in the FLD group was 1.16 (p=0.040) and 1.46 (p<0.001) in univariate and multivariate logistic regression analyses, respectively. Linear regression analyses revealed that FLI was positively associated with AHT in both univariate and multivariate analyses. Analyses using a propensity score-matched cohort showed trends similar to those using the total cohort. CONCLUSION: FLD and FLI were associated with poor hearing thresholds and HL. Therefore, active monitoring of hearing impairment in patients with FLD may be helpful for early diagnosis and treatment of HL in the general population.

Functional and metagenomic level diversities of human gut symbiont-derived glycolipids.

Bioactive metabolites produced by symbiotic microbiota causally impact host health and disease, nonetheless, incomplete functional annotation of genes as well as complexities and dynamic nature of microbiota make understanding species-level contribution in production and actions difficult. Alpha-galactosylceramides produced by Bacteroides fragilis (BfaGC) are one of the first modulators of colonic immune development, but biosynthetic pathways and the significance of the single species in the symbiont community still remained elusive. To address these questions at the microbiota level, we have investigated the lipidomic profiles of prominent gut symbionts and the metagenome-level landscape of responsible gene signatures in the human gut. We first elucidated the chemical diversity of sphingolipid biosynthesis pathways of major bacterial species. In addition to commonly shared ceramide backbone synthases showing two distinct intermediates, alpha-galactosyltransferase (agcT), the necessary and sufficient component for BfaGC production and host colonic type I natural killer T (NKT) cell regulation by B. fragilis, was characterized by forward-genetics based targeted metabolomic screenings. Phylogenetic analysis of agcT in human gut symbionts revealed that only a few ceramide producers have agcT and hence can produce aGCs, on the other hand, structurally conserved homologues of agcT are widely distributed among species lacking ceramides. Among them, alpha-glucosyl-diacylglycerol(aGlcDAG)-producing glycosyltransferases with conserved GT4-GT1 domains are one of the most prominent homologs in gut microbiota, represented by Enterococcus bgsB . Of interest, aGlcDAGs produced by bgsB can antagonize BfaGC-mediated activation of NKT cells, showing the opposite, lipid structure-specific actions to regulate host immune responses. Further metagenomic analysis of multiple human cohorts uncovered that the agcT gene signature is almost exclusively contributed by B. fragilis , regardless of age, geographical and health status, where the bgsB signature is contributed by >100 species, of which abundance of individual microbes is highly variable. Our results collectively showcase the diversities of gut microbiota producing biologically relevant metabolites in multiple layers-biosynthetic pathways, host immunomodulatory functions and microbiome-level landscapes in the host.

Kinetic and mechanistic diversity of intestinal immune homeostasis characterized by rapid removal of gut bacteria.

Symbiotic microbiota critically contribute to host immune homeostasis in effector cell-specific manner. For exclusion of microbial component, germ-free animals have been the gold standard method. However, total removal of the entire gut microbiota of an animal from birth significantly skews physiological development. On the other hand, removal of gut microbiota from conventional mice using oral antibiotics has its own limitations, especially lack of consistency and the requirement for long-term treatment period. Here, we introduce an improved regimen to quickly remove gut microbiota and to maintain sterility, that is well received by animals without refusal. Rapid and consistent exclusion of resident bacteria in the gut lumen revealed kinetic differences among colonic lymphocyte subsets, which cannot be observed with typical germ-free animal models. Furthermore, the proposed method distinguished the mechanism of microbiota contribution as a direct stimulus to capable effector cells and a homeostatic cue to maintain such cell types.

Effect of prehydration solution on hearing threshold after chemotherapy in patients with head and neck cancers: a retrospective study.

BACKGROUND: The study aimed to evaluate the effect of prehydration solution on hearing thresholds after cisplatin chemotherapy. METHODS: In this retrospective cohort study, we reviewed the data of patients who underwent ≥3 courses of cisplatin-based chemotherapy for locally advanced head and neck cancers at a tertiary referral center (n=64). The dextrose solution (DW) group (n=26) received 2 L of normal saline and 1 L of 5% dextrose. The Hartmann solution (HS) group (n=38) received 2 L of normal saline and 1 L of HS. Hearing data were measured 1 day before starting the first course of chemotherapy, and again 20 days after the first, second, and third courses of chemotherapy. The severity of hearing loss was evaluated using the Common Terminology Criteria for Adverse Events (CTCAE). RESULTS: Thresholds at all frequencies after chemotherapy were greater in the DW group than in the HS group. The increase in thresholds in 1 to 4 kHz after the third course of chemotherapy was greater in the DW group than in the HS group. CTCAE grades after the second and third courses of chemotherapy were greater in the DW group than in the HS group. Logistic regression showed that the odds ratio for CTCAE grade 3 or 4 after the third course of chemotherapy in the DW group was 4.84 on univariate analysis. CONCLUSION: Prehydration using a solution with salt was associated with a decrease in change in hearing thresholds after cisplatin chemotherapy in patients with head and neck cancers.

N-glycosylation of UNC93B1 at a Specific Asparagine Residue Is Required for TLR9 Signaling.

Toll-like receptors (TLRs) play critical roles in the first line of host defense against pathogens through recognition of pathogen-associated molecular patterns and initiation of the innate immune responses. The proper localization of TLRs in specific subcellular compartments is crucial for their ligand recognition and downstream signaling to ensure appropriate responses against pathogens while avoiding erroneous or excessive activation. Several TLRs, including TLR7 and TLR9 but not TLR4, depend on UNC93B1 for their proper intracellular localization and signaling. Accumulating evidence suggest that UNC93B1 differentially regulates its various client TLRs, but the specific mechanisms by which UNC93B1 controls individual TLRs are not well understood. Protein N-glycosylation is one of the most frequent and important post-translational modification that occurs in membrane-localized or secreted proteins. UNC93B1 was previously shown to be glycosylated at Asn251 and Asn272 residues. In this study, we investigated whether N-glycosylation of UNC93B1 affects its function by comparing wild type and glycosylation-defective mutant UNC93B1 proteins. It was found that glycosylation of Asn251 and Asn272 residues can occur independently of each other and mutation of neither N251Q or N272Q in UNC93B1 altered expression and localization of UNC93B1 and TLR9. In contrast, CpG DNA-stimulated TLR9 signaling was severely inhibited in cells expressing UNC93B1(N272Q), but not in cells with UNC93B1(N251Q). Further, it was found that glycosylation at Asn272 of UNC93B1 is essential for the recruitment of MyD88 to TLR9 and the subsequent downstream signaling. On the other hand, the defective glycosylation at Asn272 did not affect TLR7 signaling. Collectively, these data demonstrate that the glycosylation at a specific asparagine residue of UNC93B1 is required for TLR9 signaling and the glycosylation status of UNC93B1 differently affects activation of TLR7 and TLR9.

Gut Microbiota-Derived Unconventional T Cell Ligands: Contribution to Host Immune Modulation.

Besides the prototypic innate and adaptive pathways, immune responses by innate-like lymphocytes have gained significant attention due to their unique roles. Among innate-like lymphocytes, unconventional T cells such as NKT cells and mucosal-associated invariant T (MAIT) cells recognize small nonpeptide molecules of specific chemical classes. Endogenous or microbial ligands are loaded to MHC class I-like molecule CD1d or MR1, and inducing immediate effector T cell and ligand structure is one of the key determinants of NKT/MAIT cell functions. Unconventional T cells are in close, constant contact with symbiotic microbes at the mucosal layer, and CD1d/MR1 can accommodate diverse metabolites produced by gut microbiota. There is a strong interest to identify novel immunoactive molecules of endobiotic (symbiont-produced) origin as new NKT/MAIT cell ligands, as well as new cognate Ags for previously uncharacterized unconventional T cell subsets. Further studies will open an possibility to explore basic biology as well as therapeutic potential.

Internal Carotid Artery Pseudoaneurysm Secondary to Skull Base Osteomyelitis: A Case Report.

We report a case of a pseudoaneurysm secondary to skull base osteomyelitis (SBO) in an 82-year-old female. The patient was hospitalized with an acute episode of bleeding from the right ear, which persisted despite packing placed in the ear. We suspected bleeding from the internal carotid artery (ICA) and performed angiography, which revealed a pseudoaneurysm that presumably developed secondary to invasion of the wall of the petrous segment of the right ICA, and the patient underwent emergency coil embolization. Bleeding from the ear recurred a week later, and we performed repeat angiography, followed by embolization and deployment of multiple stents at the site of the pseudoaneurysm, which controlled the bleeding. Clinicians should be mindful of a pseudoaneurysm as a rare complication of SBO, following the spread of infection to adjacent soft tissues or vessels. A pseudoaneurysm should be considered in the differential diagnosis in patients with recurrent epistaxis or bleeding from the ears in addition to cranial nerve symptoms, and this condition warrants urgent evaluation.

Host immunomodulatory lipids created by symbionts from dietary amino acids.

Small molecules derived from symbiotic microbiota critically contribute to intestinal immune maturation and regulation1. However, little is known about the molecular mechanisms that control immune development in the host-microbiota environment. Here, using a targeted lipidomic analysis and synthetic approach, we carried out a multifaceted investigation of immunomodulatory α-galactosylceramides from the human symbiont Bacteroides fragilis (BfaGCs). The characteristic terminal branching of BfaGCs is the result of incorporation of branched-chain amino acids taken up in the host gut by B. fragilis. A B. fragilis knockout strain that cannot metabolize branched-chain amino acids showed reduced branching in BfaGCs, and mice monocolonized with this mutant strain had impaired colonic natural killer T (NKT) cell regulation, implying structure-specific immunomodulatory activity. The sphinganine chain branching of BfaGCs is a critical determinant of NKT cell activation, which induces specific immunomodulatory gene expression signatures and effector functions. Co-crystal structure and affinity analyses of CD1d-BfaGC-NKT cell receptor complexes confirmed the interaction of BfaGCs as CD1d-restricted ligands. We present a structural and molecular-level paradigm of immunomodulatory control by interactions of endobiotic metabolites with diet, microbiota and the immune system.

Prediction of hearing outcomes in chronic otitis media patients underwent tympanoplasty using ossiculoplasty outcome parameter staging or middle ear risk indices.

PURPOSE: Ossiculoplasty outcome parameter staging (OOPS) and middle ear risk index (MERI) are the most commonly used indices for predicting prognosis of patients with chronic otitis media (COM). This study aimed to verify the efficiency of OOPS and MERI scores in predicting outcomes of patients with COM who underwent tympanoplasty. METHODS: We retrospectively reviewed the data of patients who underwent tympanoplasty (n = 526). OOPS, and MERI scores were collected. Hearing data were measured 1 day preoperatively, and 3 and 12 months postoperatively. Operation success was defined according to the Korean Society of Otology guidelines. RESULTS: For calculation of success, the ROC values of MERI were 0.551 at 12 months. ROC values of OOPS were 0.637 at 12 months. There were no significant differences in hearing variables among the three groups according to MERI. There were significantly favorable outcomes in hearing variables in the low-risk group in OOPS. The mean OOPS score was greater in patients with success than those with non-success. Otorrhea, ossicle status, and status of mucosa as variables in both indices were associated with success. The type of mastoidectomy as a variable in OOPS alone was associated with success. Absence of hypertension, presence of ossiculoplasty, and use of incus as ossiculoplasty material were associated with poor success rate. CONCLUSION: Compared with MERI, the OOPS index was more closely associated with the hearing outcomes, which may be due to the extent of inflammation in the OOPS index.

Catalyst-controlled divergent transformations of N-sulfonyl-1,2,3-triazoles into isoquinolin-3-ones and 2-aminoindanones.

Novel catalyst-controlled divergent intramolecular reactions of N-sulfonyl-1,2,3-triazoles with tethered-allylic alcohol have been developed. In the presence of the Pd(0) catalyst alone, 1-vinylated 1,4-dihydroisoquinolin-3-ones were formed, whereas 3-vinylated 2-aminoindanones were accessed under tandem, one-pot, Rh(ii)/Pd(0) dual catalytic conditions. Based on deuterium-labelling experiments and isolation of the intermediate, a plausible reaction mechanism has been proposed.

Clinical outcomes of otogenic skull base osteomyelitis.

PURPOSE: Skull base osteomyelitis (SBO) is an uncommon and a potentially life-threatening condition if not promptly recognized and properly treated. The aim of our study was to present a 32-case series of patients diagnosed with SBO at a single center. METHODS: In this retrospective study, we reviewed the data of patients diagnosed with otogenic SBO between January 2011 and January 2020. 32 patients were enrolled in the study. SBO diagnosis was based on a combination of symptoms and physical examination, bone scan, brain magnetic resonance imaging, and pathologic examination findings. The following clinical data were collected during the follow-up period: types of antibiotics used, duration of antibiotic treatment, C-reactive protein level, presence of disease control, duration from the onset of symptoms to diagnosis, and patient survival. RESULTS: The mean follow-up period was 11 (1-110) months. The mean duration of antibiotic treatment was 115 (19-223) days. The mean C-reactive protein levels at the time of diagnosis and at the endpoint of follow-up were 3.05 (0.56-18.31) and 0.21 (0.03-33.61) mg/dL, respectively (P < 0.001). Disease control rate was 34.9% at 1-year and 83.7% at 5-year follow-up. Patient survival rate was 90.6% at 1- and 3-year follow-ups. At the endpoint of follow-up, three patients died. The mean durations from the onset of symptoms to diagnosis were 50 (5-360) and 90 (30-480) days in patients with the controlled disease and in those with the uncontrolled disease, respectively, at the endpoint of follow-up (P = 0.043). CONCLUSION: Comprehensive assessment and aggressive treatment of patients exhibiting symptoms suggestive of SBO would help in the rapid diagnosis of otogenic SBO, resulting in an improvement in prognosis.

Comparison of Ossiculoplasty Outcomes Using Different Materials in the Treatment of Chronic Otitis Media.

OBJECTIVE: This study aimed to evaluate success rates after ossicular chain reconstruction using different materials. STUDY DESIGN: Retrospective cohort study. SETTING: Tertiary referral center. PATIENTS: Four hundred forty-three participants who underwent ossiculoplasty at a tertiary medical center were included. INTERVENTION: Ossicular chain reconstruction using five materials: autologous malleus, incus, and cortical bone, as well as Hydroxyapatite (HA) and titanium. MAIN OUTCOME MEASURES: Hearing data were measured 1 day preoperatively and 6 months postoperatively. Successful hearing outcomes were defined by the fulfillment of more than one of the following criteria: postoperative air-bone gap of 20 dB or less, hearing air conduction (AC) gain of 15 dB or more, or postoperative AC hearing less than 30 dB. RESULTS: Preoperative median AC values (95% confidence interval) among participants with malleus, incus, cortical bone, HA, and titanium transplants or prostheses were 50 (39.6-54.6) dB, 51.3 (48.1-51.8) dB, 50 (45.2-52.2) dB, 56.3 (50.9-57.6) dB, and 54.3 (48.5-56.0) dB, respectively (p = 0.092). The success rates in malleus, incus, cortical bone, HA, and titanium were 53.3%, 60.3%, 51.7%, 61.6%, and 69.7%, respectively. Titanium had the highest success rate among the five materials, but the differences between the materials were not statistically significant (p = 0.283). Titanium had highest success rate among the participants with erosive stapes suprastructure or obstructed Eustachian tubes (p = 0.042 for erosive stapes suprastructure and p = 0.010 for obstructed Eustachian tubes). CONCLUSION: Our study demonstrated that titanium prostheses would be a good alternative for ossiculoplasty in cases wherein autologous material is unavailable, especially in association with unfavorable conditions, such as with the presence of cholesteatoma, erosive stapes suprastructure, edematous middle ear mucosa, and obstructed Eustachian tube.

Synergistic Dual Transition Metal Catalysis.

Synergistic catalysis, a type of plural catalysis which utilizes at least two different catalysts to enable a reaction between two separately activated substrates, has unlocked a plethora of previously unattainable transformations and novel chemical reactivity. Despite the appreciable utility of synergistic catalysis, specific examples involving two transition metals have been limited, as ensuring a judicious choice of reaction parameters to prevent deactivation of catalysts, undesirable monocatalytic event(s) leading to side products, or premature termination and other potentially troublesome outcomes present a formidable challenge. Excluding those driven by photocatalytic mechanisms, this review will highlight the reported examples of reactions that make use of two simultaneous catalytic cycles driven by two transition metal catalysts.

Flow enhances phenotypic and maturation of adult rat liver organoids.

A biologically relevant in vitro model of hepatic microtissue would be a valuable tool for the preclinical study of pharmacokinetics and metabolism. Although considerable advances have been made in recent years in the establishment of alternative in vitro culture systems that mimic liver tissue, generating an effective liver model remains challenging. Specifically, existing model systems still exhibit limited functions for hepatocellular differentiation potential and cellular complexity. It is essential to improve the in vitro differentiation of liver progenitor cells (LPCs) for disease modeling and preclinical pharmatoxicological research. Here, we describe a rat liver organoid culture system under in vivo-like steady-state flow conditions; this system is capable of controlling the expansion and differentiation of rat liver organoids over 10-15 d. LPCs cultured in medium flow conditions become self-assembled liver organoids that exhibit phenotypic and functional hepato-biliary modeling. In addition, hepatocytes that are differentiated using liver organoids produced albumin and maintained polygonal morphology, which is characteristic of mature hepatocytes.