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Investigating immunization via the esophagus:carrageenan’s impact on immune activation

Journal of Biomedical and Translational Research.2024;25(2):101-106. doi:10.12729/jbtr.2024.25.2.101

This study aimed to investigate the effectiveness of carrageenan (CGN) as an oral immune adjuvant. During the initial research, the inadvertent shallow insertion of an oral gavage nee-dle confirmed CGN’s effect as an adjuvant for esophageal immunization. However, in oral immunization, antibody formation was not observed regardless of CGN’s presence or ab-sence as an adjuvant. Conversely, when bovine serum albumin (used as an antigen) was in-troduced into the esophagus along with CGN, it resulted in the production of antigen-specific IgG. An exploration was conducted to ascertain whether CGN’s adjuvant effects were associ-ated with prolonging the antigen’s residence time in the esophagus. Upon introducing the an-tigen into the esophagus without CGN, it was undetectable at two minutes post-introduction.Conversely, when administered with CGN, the antigen remained detectable in the esophagus for up to five minutes post-introduction. To investigate whether this immune response was elicited through mucosal immune mechanisms in the esophagus, the production of IgA, a representative immunoglobulin of mucosal immunity, was assessed. Following esophageal immunization with CGN as an adjuvant, total IgG, IgG1, and IgG2a were detected in serum, while IgA was not detected. These findings suggest that under specific conditions, the esoph-agus may serve as a site for initiating a novel immune response.

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Patents trends analysis of microbiome-based treatment technology for neurological disorders in pets

Byung-Suk JEON ; Huiyeong JEONG ; Sohee JEONG ; Changjong MOON ; Jong-Hwan PARK ; Sung-Ho KIM ; Joong-Sun KIM

Journal of Biomedical and Translational Research.2024;25(2):33-39. doi:10.12729/jbtr.2024.25.2.33

This study was conducted to collect the patents of microbiome-based treatment technology for pets. An electronic search for microbiome or probiotics in brain nervous system disease was studied using the WINTELIPS database. Patent Cooperation Treaty of Korea, Japan, the EU, the US, and China that were registered by October 31, 2022 were selected in this study.A total of 206 patents were included for final analysis. Since 2016, patent activity has shown an explosive increase in recent years. China is the leading market in this technology field, and Korea has the second-highest market share. To provide the groundwork for the next research and development, we examined the industrial trend of microbiome for brain nervous system diseases in this study using an analysis of patents that have been applied for andregistered up to this point. Looking at the overall patent trends by year in the technology field related to treating of brain and nervous system diseases using the microbiome, there was a tendency to repeat increasing and decreasing trends. However, considering 2021 and 2022, which have undisclosed sections, it can be seen that patent activity has tended to increase explosively in recent years, starting in 2016. If related studies use the patent analysis data constructed in this way strategically, it is expected that it will lead to patent registration and the development of new products, ultimately contributing to the revitalization of the companion animal industry.

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Preliminary investigation on antimicrobial uses in small animal clinics in South Korea

Obaidul ISLAM ; Heyong-seok KIM ; Jong-Won KIM ; Kyung-Duk MIN

Journal of Biomedical and Translational Research.2024;25(2):89-99. doi:10.12729/jbtr.2024.25.2.89

Antimicrobial resistance significantly threatens human and animal health globally, with con-siderable mortality and economic impact. This study investigated antimicrobial usage in small animal clinics in South Korea, focusing on understanding the trends in prescriptions for therapeutic and preventive purposes. Data were collected from 12 small animal clinics that were analyzed for antimicrobial prescriptions from 2018–2020. A comprehensive dataset was used, including patient signalment, clinical notes, and prescription details, and statistically analyzed using SPSS software. The results indicated that most antimicrobials (93.1%) were prescribed for the treatment of infectious diseases, with a smaller portion (6.9%) used for pre-ventive measures, such as surgery. High prescription rates were observed for the treatment of cutaneous and otological diseases, which may reflect common diseases in companion an-imals. The study highlighted a higher prescription rate for adult age groups, possibly because of the higher prevalence in those groups. Overall, this study provides valuable insights into common prescription patterns in veterinary practice and underscores the need for more strin-gent antimicrobial stewardship to curb the rise of antimicrobial resistance. This suggests that ongoing surveillance and education on appropriate antimicrobial use are crucial for optimizing treatment outcomes and minimizing the development of resistance.

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Inflammatory bowel disease pathogenesis mediated by Th17 cells: cytokines, microbiota, and therapies

Ji-Hyun PARK ; Min Hyeok LEE ; Chan-Su PARK

Journal of Biomedical and Translational Research.2024;25(2):15-32. doi:10.12729/jbtr.2024.25.2.15

Inflammatory bowel disease (IBD) is a chronic condition characterized by continuous inflam-mation of the gastrointestinal tract that varies in intensity over time. IBD is caused by several factors including aberrant gut flora, immunological dysregulation, altered environmental con-ditions, and genetic variations. However, the pathogenesis of IBD remains unclear. Studies have indicated that an imbalance between T helper 17 (Th17) and regulatory T (Treg) cells contributes significantly to the development of IBD. Intestinal Tregs suppress inflammation and are critical for maintaining tissue homeostasis. Th17 cells are known to play an import-ant role in the development and pathogenesis of IBD and provide non-inflammatory support for the integrity of the intestinal barrier against bacterial and fungal infections. Therefore, the Th17/Treg cell balance is crucial in the pathogenesis of IBD and gut integrity. The micro-environment of the intestinal mucosal immunity is regulated by the secretion of cytokines associated with Th17 cells and Tregs. Several studies have indicated that the gut bacteria contribute to the control of the immune milieu and play a key role in the regulation of Th17 cell development. Intestinal bacteria and cytokines control Th17 cell development. Th17 cells secrete cytokines that regulate the immune microenvironment in the gut mucosa. This review provides an overview of Th17 cells and examines the strategies for treating patients with IBD using Th17 cell-targeted drugs.

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Potential use of ashwagandha extract in storage of liquid boar semen

Malavi Arachchi Gamage Nayodya Dananjanee Malavi Arachchi GAMAGE ; Wijesooriya Mudhiyanselage Nadeema DISSANAYAKE ; Young-Joo YI

Journal of Biomedical and Translational Research.2024;25(2):79-88. doi:10.12729/jbtr.2024.25.2.79

The functional roles of plant extracts have been investigated for the treatment of variousdiseases including subfertility. Recent studies have highlighted the benefits of ashwagandha extract (AE) in enhancing sperm production, boosting testosterone levels, and lowering re-active oxygen species (ROS) levels in mammals. The current study is to examine the effectsof the addition of AE to liquid boar semen on sperm quality during storage and its potentialapplication in assisted reproductive technology. A hot water extract of ashwagandha was prepared from the dried powder of ashwagandha roots. Boar spermatozoa were stored in Beltsville thawing solution (BTS) at 17℃ for 5 days, with various concentrations of AE (1–50 mg/mL). During storage, motility, viability, acrosomal integrity and ROS of boar spermatozoa were examined. The results have shown that sperm stored in BTS with varying quantities of AE ranging from 1–20 mg/mL exhibited higher motility compared to those without AE (control) or with 50 mg/mL AE for 5 days. Similarly, sperm viability was better maintained in sperm treated with 1–20 mg/mL AE. Moreover, sperm stored in BTS with AE led to significantly high-er acrosomal integrity and chromatin stability rates than sperm stored without AE. Notably, intracellular ROS levels significantly decreased in sperm stored in BTS with AE. Particularly, spermatozoa stored at 10 mg/mL AE exhibited an effective reduction in ROS during storage. These findings suggest the potential role of AE as an additive during sperm storage maintains sperm quality and can be used during subfertility treatment in both animals and humans.

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Alteration of goat kids’ gut microbiota by Cryptosporidium parvum infection

Subin LEE ; Badriah ALKATHIRI ; Ah-Young KIM ; Kyung-Hyo DO ; Kwangwon SEO ; Soochong KIM ; Wan-Kyu LEE ; Dongmi KWAK ; Seung-Hun LEE

Journal of Biomedical and Translational Research.2024;25(2):41-52. doi:10.12729/jbtr.2024.25.2.41

Caprine cryptosporidiosis mainly occurs in young goats, with morbidity rates of 80%–100% and mortality over 50% in goat kids. However, limited research has been conducted on the impact of Cryptosporidium parvum, a diarrhea-causing pathogen, on the intestinal microbiota of goat kids. In this study, 16S rRNA-based metataxonomic analysis was performed to compare the microbial diversity and abundance of the gut microbiota between C. parvum-infected and uninfected goat kids. In total, 12 goat fecal samples were collected, including seven naturally C. parvum-infected and five uninfected goats from Chungcheongbuk-do, Korea. After amplification of the V3–V4 hypervariable region of the bacterial 16S rRNA, high-throughput sequencing was performed. The results showed differences in the microbial composition between C. parvum-infected and uninfected groups based on beta diversity. Firmicutes and Bacteroidetes were the most dominant phyla in both groups. However, no significant differ-ence was observed in the Bacteroidetes/Firmicutes ratio between the two groups. Compared with the uninfected group, the C. parvum-infected group showed significantly higher abun-dances of Tyzzerella nexillis, Lactobacillus johnsonii, Butyricicoccus pullicaecorum, Entero-coccus raffinosus, Enterococcus faecalis, and Negativicoccus massiliensis, and significantly reduced abundances of Aerococcus vaginalis, Faecalicoccus pleomorphus, Oribacterium parvum, and Coprococcus comes. These findings indicate that C. parvum infection, which is associated with diarrhea in neonatal goats, induces alterations in the caprine gut microbiota.

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Single dose oral toxicity study of a water extract of Gekko gecko in Sprague-Dawley rats

Byung-Suk JEON ; Hyeon Hwa NAM ; Huiyeong JEONG ; Joong-Sun KIM ; Sueun LEE ; Ji Hye LEE ; Yun-Soo SEO

Journal of Biomedical and Translational Research.2024;25(2):69-77. doi:10.12729/jbtr.2024.25.2.69

Traditional medicine and herbal remedies are gaining popularity worldwide, comprising a significant portion of healthcare research, advancements, and market demand. Growing scientif-ic evidence supports their substantial efficacy as pharmaceutical ingredients and dietary sup-plements in preventive healthcare. When developing pharmaceuticals, it is crucial to ensure that ingredients are free from side effects and toxicity in order to prioritize safety. Geckos, known as shou gong, are a diverse group of lizards that are widely utilized for treating various diseases in Korean Medicine. This study was conducted to assess the potential acute toxicity of a water extract Gekko gecko by a single oral dose in Sprague-Dawley rats. Twenty rats of each sex were randomly assigned to four groups (5 rats each). Test articles were administrat-ed once by oral gavage to rats at dose levels of 0, 500, 1,000, or 2,000 mg/kg body weight. Mortality, changes of body weight, and clinical signs of gross observation were monitored for 14 days after dosing. At the end of a 14-day observation period, all animals were sacrificed and complete macroscopic and hematological examinations were performed. There was no dead animal or test article-related effect on clinical signs, body weight, or gross finding. Other specific changes were not found between control and treated groups in hematology. Results showed no adverse effect at a dose of 500, 1,000, or 2,000 mg/kg in rats. The minimal lethal dose was considered to be over 2,000 mg/kg body weight in rats.

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Comparison of in vitro models for drug-induced liver injury assessment

Nam-Ju KIM ; Ji-Hyun BANG ; Hee YI ; Hyun-Ok KU ; Joong-Sun KIM ; Ji-Yeon KIM ; Byung-Suk JEON

Journal of Biomedical and Translational Research.2024;25(2):53-67. doi:10.12729/jbtr.2024.25.2.53

Drug-induced liver injury (DILI) is considered to be a significant cause of drug wastage. Tomitigate clinical DILI risks, assessing drugs using human liver models is crucial since animal studies may fall short due to species-specific liver pathway variations. Cell-based preclinical hepatotoxicity testing is often pertinent. In the present study, cells from a human liver cancer line (HepG2 and HepaRG) were cultured in both formats of 2D and 3D spheroids to explore their responses to drugs. Liver-specific marker expressions across cell lines and culture formats were also examined to assess disparities in DILI marker expressions. After treating each cell with the drugs, cytotoxicity and liver injury markers aspartate aminotransferase and alanine aminotransferase were increased. In addition, liver specific markers albumin and urea decreased in a drug concentration-dependent manner. These findings were consistent with drug sensitivity. Additionally, mRNA expression levels of cytochrome P450 enzymes (CYPs) involved in hepatocellular drug metabolism were compared following treatment with enzyme inducers. CYP1A2 and CYP2C9 were not epxressed in HepG2 cells. HepaRG cells exhibited significantly increased expression of CYP1A2, 2C9, and 3A4 post-treatment. No-tably, enzyme expression was notably higher in 3D cultures than in 2D cultures. Collectively, these findings suggest that HepaRG cells and 3D cultures hold promise for evaluating DILI during early-stage drug development.

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Ex vivo culture conditions for bioluminescence imaging of porcine skin tissue

Mina KIM ; Hao JIN ; Yong Jin LEE ; Sang-Myoeng LEE

Journal of Biomedical and Translational Research.2024;25(1):1-9. doi:10.12729/jbtr.2024.25.1.1

In response to the expanding landscape of the biotechnology industry and the increasing demand for comprehensive drug development as well as the conduct of preclinical and clinical trials, there is a growing need for employment of diverse animal models, including both small and large animals. The focus of this study was on refining ex vivo culture techniques for bioluminescence imaging following administration of intradermal injections in large animals. To examine the feasibility of our approach, varying concentrations of the rFluc protein were administered to rats and live imaging was employed to validate the corresponding levels of expression. Subsequently, following administration of rFluc to mini-pigs, ex vivo analyses were performed on sample tissues to assess the levels of protein expression across different concentrations. In particular, optimal culturing conditions that facilitated the sustained expres-sion of the protein in samples post-euthanasia were identified. Moreover, by employing small animal imaging devices, we were able to capture clear images of the sample plates, which provided evidence of the successful application of our experimental techniques. The findings from this research represent a significant effort toward refining bioluminescence imaging methods tailored for use with large animal models—an imperative facet of contemporary drug development and biomedical research.

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Histopathological characteristics of progressive multifocal leukoencephalopathy in a patientwith acquired immune deficiency syndrome (AIDS)

Hyo Jong KIM ; Chang Gok WOO

Journal of Biomedical and Translational Research.2024;25(1):11-14. doi:10.12729/jbtr.2024.25.1.11

Progressive multifocal leukoencephalopathy (PML) is a demyelinating disease by John Cunningham virus (JC virus) infection in oligodendrocytes. The radiographic and clinical features,along with the identification JC in cerebrospinal fluid polymerase chain reaction, are sufficient for the diagnosis of PML in immunodeficiency. However, it is difficult to suspect PML without the patient history of immunodeficiency. A 32-year-old man presented with headache for a month without any medical history. Based on clinical and image features, the differential di-agnoses included demyelinating lesion and neoplasms. Microscopically, biopsy specimen showed multifocal demyelinating and degenerative white matter, consistent with PML. Oligodendrocytes cells with increased nuclei and plum-colored inclusions were admixed withperivascular lymphocytic and histiocytic infiltration, and loss of myelin. Atypical astrocytes had large or multiple nuclei. After brain biopsy, human immunodeficiency virus infection was confirmed by serum chemiluminescent immunoassay. It is unlikely that PML would be con-sidered without the information of immunosuppression. Therefore, it is very important to be aware of the histological features of PML.

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