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Abstract Some drugs or chemicals exhibit different safety profiles in newborns/young children compared to adults. Polyhexamethyleneguanidine phosphate (PHMG-P) has been implicated in the humidifier disinfectant tragedy in 2011. There are limited reports on the toxicity of PHMG-P in neonatal animals. This study aimed to assess the toxicity of PHMG-P in neonates and to compare toxicity between young and adult mice. Mice aged 7–10 days and 8 weeks were anesthetized with isoflurane and then intranasally instilled with 0.9 mg/kg and 1.5 mg/kg PHMG-P once weekly for 4 weeks. The control group was given a corresponding volume of saline intranasally. Approximately 20 h after the 4th instillation, all mice (juveniles aged 28‒31 days and adults aged 11 weeks) were euthanized. Assessments included body weights, organ weights, cytokine production, and histopathological examinations. Both juvenile and adult mice exhibited significant pulmonary toxicity. There were no significant changes in body weight in either male or female juveniles, whereas adult mice experienced 5.0‒22.2% weight loss. However, lung weights increased in both age groups, accompanied by rises in cytokines and chemokines. Histopathological analyses revealed significant lung changes in both juvenile and adult mice, including immune cell infiltration, foamy macrophage, and granulomatous inflammation. PHMG-P is known to cause inflammation and fibrotic changes in rodents and humans that persist even during long recovery periods. Further research is required to explore the long-term health effects of PHMG-P following repeated early-life exposure.

Fish surveys were conducted near artificial structures installed in eight streams flowing from the Nakdong River region to the east coast of Korea. As a result of the survey, a total of 20,000 individuals belonging to nine orders, 17 families, and 47 species were found. The dominant species was Zacco koreanus, and the subdominant species was Pungtungia herzi. Among the surveyed fish species, endangered species were Pseudobagrus brevicorpus (rank I) and Cottus hangiongensis (rank II). In addition, migratory fish species were Oncorhynchus keta, Plecoglossus altivelis, and Tribolodon hakonensis. As a result of community analysis, the highest diversity, dominance, richness, and evenness indices were Wangpi, Chuksan, Gokgang, and Janggi Stream respectively. The highest number of species was surveyed in Gokgang Stream (25 species). Alpha diversity, the number of species at each survey point in each stream, was the highest at a survey point in Wangpi Stream (20 species), and beta diversity, species at each stream survey point that does not overlap with nearby survey point, was the highest at survey points in Gokgang Stream (12 species). As a result of the similarity analysis, surveyed streams were divided into four groups: group 1 (Chuksan, Baeknok Stream), group 2 (Uljinnamdae, Wangpi, Yungdeokosip Steam), group 3 (Gokgang Stream), and group 4 (Janggi, Hoeya Stream). This study is expected to serve as basic data for managing the aquatic ecosystem of streams flowing from the Nakdong River region to the east coast.

Abstract Tuberculosis is a highly contagious disease caused by Mycobacterium tuberculosis (Mtb), which is one of the prominent reasons for the death of millions worldwide. The bacterium has a substantially higher mortality rate than other bacterial diseases, and the rapid rise of drug-resistant strains only makes the situation more concerning. Currently, the only licensed vaccine BCG (Bacillus Calmette–Guérin) is ineffective in preventing adult pulmonary tuberculosis prophylaxis and latent tuberculosis re-activation. Therefore, there is a pressing need to find novel and safe vaccines that provide robust immune defense and have various applications. Vaccines that combine epitopes from multiple candidate proteins have been shown to boost immunity against Mtb infection. This study applies an immunoinformatic strategy to generate an adequate multi-epitope immunization against Mtb employing five antigenic proteins. Potential B-cell, cytotoxic T lymphocyte, and helper T lymphocyte epitopes were speculated from the intended proteins and coupled with 50 s ribosomal L7/L12 adjuvant, and the vaccine was constructed. The vaccine’s physicochemical profile demonstrates antigenic, soluble, and non-allergic. In the meantime, docking, molecular dynamics simulations, and essential dynamics analysis revealed that the multi-epitope vaccine structure interacted strongly with Toll-like receptors (TLR2 and TLR3). MM-PBSA analysis was performed to ascertain the system’s intermolecular binding free energies accurately. The immune simulation was applied to the vaccine to forecast its immunogenic profile. Finally, in silico cloning was used to validate the vaccine’s efficacy. The immunoinformatics analysis suggests the multi-epitope vaccine could induce specific immune responses, making it a potential candidate against Mtb. However, validation through the in-vivo study of the developed vaccine is essential to assess its efficacy and immunogenicity profile, which will assure active protection against Mtb.

The lateral connectivity of Suncheon’s Dong stream was evaluated based on landside, waterside, and levee data of the river. A total of six items was evaluated, crosssectional structure, vegetation cover ratio, connecting waterway gate, levee slopes, urbanized area ratio and wetland area ratio. Among the items, the connectivity was high in the following order: cross-sectional structure, urbanization area ratio, and levee slope. In a total of 16 survey areas of Suncheon’s Dong stream, one, 12, and three areas were evaluated as continuity, damaged, and discontinuity respectively. The downstream section was evaluated as one continuity and three damaged, the lower-middle stream section was evaluated as one damaged and three discontinuity, and the upper-middle stream and upstream sections were evaluated as eight damaged. The cross-sectional structure, vegetation cover ratio, levee slope, and urbanized area ratio had low lateral connectivity in the lower-middle stream survey section, while the wetland area ratio had high lateral connectivity in the downstream survey section. The lateral connectivity evaluation results of the river aquatic ecosystem in Suncheon’s Dong stream can be used as basic data that is helpful in future ecological river management policies.

IntroductionDecreasing rates of blood donation and close margins between blood supply and demand pose challenges in healthcare. Genetically engineered pig red blood cells (pRBCs) have been explored as alternatives to human RBCs for transfusion, and triple-gene knockout (TKO) modification improves the compatibility of pRBCs with human blood in vitro. In this study, we assessed the efficacy and risks of transfusing wild-type (WT)- and TKO-pRBCs into nonhuman primates (NHPs).MethodsBlood from O-type WT and TKO pigs was processed to produce pRBCs for transfusion, which were transfused or not into NHPs (n=4 per group: WT, TKO, and control) after 25% total blood volume withdrawal: their biological responses were compared. Hematological, biochemical, and immunological parameters were measured before, immediately after, and at intervals following transfusion. Two months later, a second transfusion was performed in three NHPs of the transfusion group.ResultsTransfusion of both WT- and TKO-pRBCs significantly improved RBC counts, hematocrit, and hemoglobin levels up to the first day post-transfusion, compared to the controls. The transfusion groups showed instant complement activation and rapid elicitation of anti-pig antibodies, as well as elevated liver enzyme and bilirubin levels post-transfusion. Despite the higher agglutination titers with WT-pRBCs in the pre-transfusion crossmatch, the differences between the WT and TKO groups were not remarkable except for less impairment of liver function in the TKO group. After the second transfusion, more pronounced adverse responses without any hematological gain were observed.ConclusionsWT- and TKO-pRBC transfusions effectively increased hematologic parameters on the first day, with rapid clearance from circulation thereafter. However, pRBC transfusion triggers strong antibody responses, limiting the benefits of the pRBC transfusion and increasing the risk of adverse reactions.

The vertical connectivity of the river aquatic ecosystem was evaluated based on fish and artificial structures in Dorim stream, an urban stream in Seoul. As a result of a survey in the downstream area in 100.0 m of a total of 71 artificial structures, 13,728 individuals of fishes belonging to five orders, seven families, and 25 species were investigated, with the dominant species Zacco platypus and the subdominant species Rhynchocypris oxycephalus. As for endemic species, seven species were investigated and in terms of feeding characteristics, omnivorous species were the most common with 17 species (68%). Also an alien species, Poecilia reticulata was found. Fish species tended to decrease as the survey was conducted to upstream. Based on the movement characteristics of the fish species and the features of artificial structure survey results, the longitudinal continuity of each artificial structure was evaluated as 43 continuity, two damaged, 19 discontinuity, and seven absent. In inclined structures, stream velocity was found to be the main factor for discontinuity. In vertical structures, the down depth and head drop appeared to be the main factors for discontinuity. The results of this survey are expected to serve as basic data for the conservation of river aquatic ecosystems in the future.

Herbaceous strata with a dominant plant community 50 m in size were selected from 10 reservoir embankment areas in Shinan-gun, a southwestern island region of South Korea. Terrestrial insects were sampled 100 times with quantitative sweep sampling to investigate the insect fauna and their association with flora. The survey areas were mainly dominated either by Phragmites japonica or Miscanthus sinensis plant species. A total of 1,948 individual insects representing 173 species from 64 families within 9 orders were collected. The number of species was the highest at Jaeundo while the Shannon-Wiener diversity index was the highest at Saokdo. Similarity analysis showed that the insect species composition tended to be similar among survey sites with plant communities comprising the same dominant plant species. Species diversity was the highest in April while the number of individual insects was the highest in September. Petaphora maritima and Ochetellus glaber were the dominant and subdominant insect species, with a high frequency of occurrence at these sites. The results of this study could be applied towards biodiversity conservation and monitoring of reservoir embankment areas in the future.

The purpose of this study was to develop a monoclonal antibody (mAb) that can identify porcine dendritic cells (DCs) that have differentiated from bone marrow progenitor cells. Hybridoma technology was used to obtain mAbs, and bone marrow-derived DCs (BMDCs) were employed as immunogens for producing antibodies. The generated PD9-9 mAbs exhibited considerable reactivity towards porcine BMDCs with applications in flow cytometry and immunostaining. The antibody was composed of heavy immunoglobulin gamma-1 chains and light kappa chains. The PD9-9 mAb recognized fully differentiated porcine BMDCs and cells undergoing DC differentiation. In contrast, bone marrow cells and macrophages were not recognized by PD9-9. In addition, the PD9-9 mAb promoted porcine DC proliferation. Consequently, the PD9-9 mAb may be a biomarker for porcine DCs and will be advantageous for investigating porcine DC biology.

Because there is a shortage of donor kidneys, researchers are exploring the possibility of using genetically modified pig kidneys for transplantation. Approaches involving knockout of carbohydrate genes or knockin of protective proteins have been attempted to determine the best gene modifications. In this study, we utilized GalT −/− ; hCD39;hCD55 and GalT −/− ; hCD39;hCD46;hCD55;thrombomodulin (TBM) pigs for transplantation in nonhuman primates (NHPs). The NHPs survived for 4 weeks after kidney transplantation (4 WAT) from the GalT −/− ; hCD39;hCD55 pig and for 6 WAT from the GalT −/− ; hCD39;hCD46;hCD55;TBM pig. However, messenger RNA (mRNA) sequencing and immunohistochemistry analysis revealed that the 6 WAT kidney exhibited more severe apoptosis, inflammation, loss of renal function, and renal fibrosis than the 4 WAT kidney. These results indicate that additional knockin of complement regulator ( hCD46 ) and coagulation regulator ( TBM ) is not enough to prevent renal damage, suggesting that improved immune suppression is needed for more prolonged survival.

IntroductionA reliable standard model is required to evaluate the efficacy of new drugs for companion animals, especially dogs. Canine atopic dermatitis (cAD), also known as allergic inflammatory skin disease, is a common condition. Currently, the house dust mite animal model is used in the research of cAD; however, this model exhibits significant individual variation and is difficult to standardize. In this study, we used ovalbumin as an antigen to sensitize and stimulate dogs, thereby establishing a stable model mimicking the T-helper 2 (Th2) response seen in cAD. Our objective was to create a cAD model that could be employed to evaluate the efficacy of novel drugs and mimic the Th2 dominant allergic response observed in the pathogenesis of atopic dermatitis of dogs.MethodsIn this study, six beagles were used. Normal saline was applied to two animals, and ovalbumin to four, on their dorsal skin.ResultsThe ovalbumin-treated groups exhibited clinical cAD symptoms, such as pruritus and erythema. Moreover, plasma levels of the cAD markers immunoglobulin E and CCL17 chemokine were higher in the ovalbumin-treated group than in the vehicle control group. The skin thickness of the epidermis was significantly increased in the ovalbumin-treated group, with infiltration of inflammatory cells observed in the thickened dermis region. In conclusion, treatment of canine skin with an optimal concentration of ovalbumin induced typical cAD-like symptoms, and histological and molecular analyses confirmed an enhanced Th2-related immune response.ConclusionTherefore, we successfully established a suitable Th2-dominant response mimicking cAD, which will facilitate targeted research of atopic dermatitis in dogs.

IntroductionThe global shortage of human blood for medical use has prompted the development of alternative blood sources. Nonhuman primates (NHPs) are commonly used owing to their physiological similarities to humans. The objective of the current study was to establish a controlled-blood-loss model in NHPs to explore their clinical and biological responses.MethodsBlood was sequentially withdrawn from 10 cynomolgus monkeys (10, 14, 18, 22, and 25% of the total blood volume); their vital signs were monitored, and blood parameters were serially analyzed. Humoral mediators in the blood were measured using flow cytometry and enzyme-linked immunosorbent assays.ResultsIn NHPs subjects to 25% blood loss and presenting with related clinical symptoms, the systolic blood pressure ratio on day 0 after bleeding was significantly lower than that of the animals from the other groups (median: 0.65 vs. 0.88, P = 0.0444). Red blood cell counts from day 0–14 and hematocrit levels from day 0–7 were markedly decreased relative to the baseline (P < 0.01). These parameters showed a direct correlation with the extent of blood loss. The levels of creatine phosphokinase, aspartate aminotransferase, and alanine aminotransferase exhibited increases in response to blood loss and had a stronger correlation with the hemoglobin ratio than the volume of blood loss. The levels of C3a and C4a, as well as interleukin (IL)-1α and IL-15, displayed a strong correlation, with no apparent association with blood loss.ConclusionThe findings of the present study showed that only NHPs with 25% blood loss exhibited clinical decompensation and significant systolic blood pressure reduction without fatalities, suggesting that this level of blood loss is suitable for evaluating blood transfusion efficacy or other treatments in NHP models. In addition, the ratio of hemoglobin may serve as a more dependable marker for predicting clinical status than the actual volume of blood loss. Thus, our study could serve as a basis for future xenotransfusion research and to predict biological responses to massive blood loss in humans where controlled experiments cannot be ethically performed.

The development of a cost-effective and accurate model for predicting the fatigue life of materials is essential for designing thermal power plants and assessing their structural reliability under operational conditions. This paper reports a novel energy-based approach for developing unified models that predict the fatigue life of boiler tube materials in ultra-supercritical (USC) power plants. The proposed method combines the Masing behavior with a cyclic stress–strain relationship and existing stress-based or strain-based fatigue life prediction models. Notably, the developed models conform to the structure of the modified Morrow model, which incorporates material toughness (a temperature compensation parameter) into the Morrow model to account for the effects of temperature. A significant advantage of this approach is that it eliminates the need for tensile tests, which are otherwise essential for assessing material toughness in the modified Morrow model. Instead, all material constants in our models are derived solely from fatigue test results. We validate our models using fatigue data from three promising USC boiler tube materials—Super304H, TP310HCbN, and TP347H—and their welded joints at operating temperatures of 500, 600, and 700 °C. The results demonstrate that approximately 91% of the fatigue data for all six materials fall within a 2.5× scatter band of the model’s predictions, indicating a high level of accuracy and broad applicability across various USC boiler tube materials and their welded joints, which is equivalent to the performance of the modified Morrow model.

Abstract Background Bentonite, a montmorillonite clay, has been used as a classical remedy strategy for a long time. Recently, bentonite has been used as a raw material in cosmetic products because of its antibacterial and antioxidant properties. However, the therapeutic effect of bentonite on burn injuries has not yet been identified. Objectives The aim of this study was to explore the therapeutic effect of a novel bentonite complex, which was developed for medical use, on burn wounds and the anti‐inflammatory function of bentonite clay in the complex in vitro. Methods A novel bentonite complex and bentonite clay were prepared for each in vivo and in vitro assay (C&L Biotech, Seoul, Korea). Burn wounds were induced on the dorsal skin of two Yucatan minipigs, and effects of tissue regeneration and anti‐inflammation were assessed by histological and gene expression analysis. Results The bentonite complex improved skin regeneration in burn wounds by inducing collagen synthesis, cell proliferation and angiogenesis in vivo. Furthermore, expression of inflammatory cytokines was significantly inhibited by treatment of the bentonite complex. In vitro study for identification of anti‐inflammatory effect showed that bentonite clay significantly regulated COX‐2 signalling in both HacaT keratinocytes and 3D4/2 macrophage cell lines. Conclusions This study identified the therapeutic effect of a novel bentonite complex in burn wounds by inducing skin regeneration and bentonite‐mediated anti‐inflammatory responses.

Abstract Background Given its similar structure and immune response to the human skin, porcine is a good model for dermal studies. Here, we sensitized ovalbumin (Ova) on minipig back skin for 2–4 weeks to induce chronic atopic dermatitis (AD). Results Gross observation, serum cytokine level, epidermal thickness, and epidermal integrity did not change after 4 weeks of Ova induction compared with the control, indicating AD modeling failure. Only the neutrophils in the blood and macrophages in bronchoalveolar lavage fluid changed slightly until 3 or 2 weeks after Ova sensitization, respectively. The successful and failed Ova-induced AD minipig models only differ in age and body weight of the minipigs. The minipigs, 12 months old with a 30-kg median weight, had a two-fold thicker dermis than minipigs 8–10 months old, with an 18.97-kg median weight, resulting in impaired Ova permeability and immune response. Conclusion Age and body weight are key factors that should be considered when developing an Ova-induced AD minipig model.

Abstract Background Quantitative real time PCR (qPCR) is a powerful tool to evaluate mRNA expression level. However, reliable qPCR results require normalization with validated reference gene(s). In this study, we investigated stable reference genes in seven tissues according to four developmental stages in minipigs. Six candidate reference genes and one target gene (ACE2) were selected and qPCR was performed. BestKeeper, geNorm, NormFinder, and delta Ct method through the RefFinder web-based tool were used to evaluate the stability of candidate reference genes. To verify the selected stable genes, relative expression of ACE2 was calculated and compared with each other. Results As a result, HPRT1 and 18S genes had lower SD value, while HMBS and GAPDH genes had higher SD value in all samples. Using statistical algorithms, HPRT1 was the most stable gene, followed by 18S, β-actin, B2M, GAPDH, and HMBS. In intestine, all candidate reference genes exhibited similar patterns of ACE2 gene expression over time, whereas in liver, lung, and kidney, gene expression pattern normalized with stable reference genes differed from those normalized with less stable genes. When normalized with the most stable genes, the expression levels of ACE2 in minipigs highly increased in intestine and kidney at PND28, which is consistent with the ACE2 expression pattern in humans. Conclusions We suggest that HPRT1 and 18S are good choices for analyzing all these samples across the seven tissues and four developmental stages. However, this study can be a reference literature for gene expression experiments using minipig because reference gene should be validated and chosen according to experimental conditions.

The microstructure of TP310HCbN welded joints and its effect on the tensile and low-cycle fatigue properties were investigated in the temperature range 20–700 °C, particularly in the operating range of 500–700 °C. The results showed that the welded joint was fully austenitic; however, a significantly different microstructure was formed in each welding zone, inducing material inhomogeneity. The material inhomogeneity was most pronounced in the weld metal, wherein an austenitic columnar grain structure composed of dendrite cores and interdendritic regions was developed. A combined analysis of the nanoindentation, failure mechanism, and kernel average misorientation using electron backscatter diffraction revealed that the dendrite core was the softest region in the welded joint and the deformation was localized here, thereby serving as a crack nucleation site and propagation path. This promoted premature failure of the welded joint, resulting in a reduction in the tensile strength, ductility, and the fatigue resistance, in comparison to the base metal. As both the welded joint and base metal showed significant cyclic hardening behavior (more than two-fold increase in the tensile peak stress) during fatigue deformation, the plastic strain energy density was found to be a suitable fatigue parameter, which was invariant throughout the fatigue life. An energy-based unified fatigue life prediction model using material toughness was developed, and its validity was demonstrated by successfully predicting the temperature dependence of the fatigue life as well as the fatigue life reduction of the welded joint.

Abstract The use of minipigs (Sus scrofa) as a platform for toxicological and pharmacological research is well established. In the present study, we investigated the effect of formaldehyde (FA) exposure on helper T cell-mediated splenic immune responses in Yucatan minipigs. The minipigs were exposed to different inhaled concentrations of FA (0, 2.16, 4.62, or 10.48 mg/m3) for a period of 2 weeks. Immune responses elicited by exposure to FA were determined by assessing physiological parameters, mRNA expression, and cytokine production. Additionally, the distribution of helper T cells and regulatory T (Treg) cells and expression of NFAT families, which are well-known T cell receptor signalling proteins associated with regulatory T cell development, were evaluated. Exposure to FA suppressed the expression of genes associated with Th1 and Th2 cells in minipigs in a concentration-dependent manner. The subsequent production of cytokines also declined post-FA exposure. Furthermore, exposure to FA induced the differentiation of CD4+ Foxp3+ Treg cells with divergent expression levels of NFAT1 and NFAT2. These results indicated that exposure to FA increased the Treg cell population via the NFAT-mediated T cell receptor signalling pathway, leading to suppression of effector T cell activity with a decline in T cell-related cytokine production.

Abstract Background Rheumatoid arthritis (RA) is a chronic inflammatory disease of joint, but there is no known cure. 3′-sialyllactose (3′-SL) is an oligosaccharide that is abundant in breast milk of mammals, and has anti-inflammatory properties. However, the efficacy of 3′-SL on RA remains unclear. The objective of the present study was to evaluate the therapeutic effect of 3′-SL after it was directly injected into the knee joint cavity of a RA minipig model. Results Minipig RA model was induced by intra-articular injection of bovine type II collagen emulsified with complete or incomplete Freund’s adjuvant into left knee joint. In clinical assessment, lameness and swelling of the hindlimb and increased knee joint width were observed in all animals. After the onset of arthritis, 3′-SL (0, 2, 10, and 50 mg/kg) was directly administered to the left knee joint cavity once a week for 4 weeks. Compared to the vehicle control group, no significant difference in macroscopic observation of the synovial pathology or the expression of inflammation-related genes (IL-1β, TNF-α, and COX2) in the synovial membrane of the knee joint was found. In microscopic observation, cell cloning of the articular cartilage was significantly reduced in proportion to the concentration of 3′-SL administered. Conclusions Our results suggest that intra-articular injected 3′-SL had a therapeutic effect on collagen-induced arthritis at the cellular level with potential as a medication for RA.

This study was conducted on the association of terrestrial insect herbivore communities with flora and plant communities in the riparian zone of an urban river in Daejeon, Korea. The survey was conducted in May, August, and October on three sites that are composed typical habitat diversity in the river. As a result of the analysis, the highest matching rate of insect herbivore host plant and flora of the site was site 1 with 93%. In addition, the dominant and subdominant species of the terrestrial insect herbivores showed a very close association with the dominant plant species that make up the plant community. As a result of analysis of similarity among the sites, the most heterogeneous group of terrestrial insect herbivore communities were in May, which presumed to be affected by changes in biomass of host plants before and after the rainy season. The results of this study can be used as data necessary for the conservation of biodiversity in riparian zone in the future.

Diclofenac sodium salt (DSS) is a widely used nonsteroidal anti-inflammatory drug. The present study was performed under good laboratory practice (GLP) regulations to investigate the toxicity of DSS after 4 weeks of repeated intramuscular administration at doses of 0, 2, 10, or 20 mg/kg/day in 32 minipigs and to evaluate the DSS effect following a 2-week recovery period. Dose-related clinical signs and alterations of hematological or clinical chemistry parameters, organ weight, and macroscopic as well as histopathological findings in hepatic, renal, gastrointestinal, skin and injection sites were observed in both sexes’ animals of the 10 or 20 mg/kg/day group. With the exception of the skin-related findings, most symptoms showed a tendency to resolve after the 2-week recovery period. The systemic exposure (AUClast) of DSS in plasma showed similar pattern to the increase rate of the dose and similar values between males and females except for the female 20 mg/kg dose group (56 %) on Day1. The systemic exposure showed a decreasing trend in the 10 or 20 mg/kg group after 4-week of repeated administration compared to Day1. The no-observed-adverse-effect level of DSS in this study was considered to be 2 mg/kg/day in both male and female minipigs.

Abstract Recently, minipig has been considered as an animal model that is appropriate for human disease model to study toxicology, pharmacology, and xenotransplantation. Nevertheless, minipigs are bred in various environment according to their use. Here, we suggest that minipigs used for research should be bred in well-controlled facility, comparing immune status of pigs raised in different breeding environment. DNA microarray was performed with ear skin and placenta of Landrace domestic pigs (DPs) and Minnesota germ-free minipigs (GPs). Their immune transcriptome was analyzed by gene ontology (GO) annotation database, based on criteria of |log2 fold change| ≥1 with P ≤ 0.05. As a result, we found that immune related genes in the ear skin of DPs were highly activated, compared to GPs. On the other hand, no significant s were found in the placenta. Quantitative real-time PCR (qRT-PCR) was performed in five candidate immune genes. Their fold changes were consistent with the results from DNA microarray (P ≤ 0.05). In conclusion, we experimentally proved that porcine immune system was affected by different breeding environment, suggesting the importance of controlling microbes in animal room for the qualified research.

Monocytes and macrophages that originate from common myeloid progenitors perform various crucial roles in the innate immune system. Stimulation with LPS combined with TLR4 drives the production of pro-inflammatory cytokines through MAPKs and NF-κB pathway in different cells. However, the difference in LPS susceptibility between monocytes and macrophages is poorly understood. In this study, we found that pro-inflammatory cytokines—IL-1β, IL-6 and TNFα showed greater induction in phorbol-12-myristate-13-acetate (PMA)-differentiated THP-1 cells than in THP-1 cells. To determine the difference in cytokine expression, the surface proteins such as TLR4-related proteins and intracellular adaptor proteins were more preserved in PMA-differentiated THP-1 cells than in THP-1 cells. MyD88 is a key molecule responsible for the difference in LPS susceptibility. Moreover, MAPKs and NF-κB pathway-related molecules showed higher levels of phosphorylation in PMA-differentiated THP-1 cells than in THP-1 cells. Upon MyD88 depletion, there was no difference in the phosphorylation of MAPK pathway-related molecules. Therefore, these results demonstrate that the difference in LPS susceptibility between THP-1 cells and PMA-differentiated THP-1 cells occur as a result of gap between the activated MAPKs and NF-κB pathways via changes in the expression of LPS-related receptors and MyD88.

Four carabid beetles, Holcoderus smaragdinus Andrewes, 1926, Lachnothorax biguttatus Motschulsky, 1862, Callistomimus chalcocephalus quadristigma (Bates, 1892), and Platyrhopalopsis picteti (Westwood, 1874) are newly recorded from Cambodia. Diagnostic characters, some collecting information, and photographs of each species are provided. Keywords: Carabidae, Coleoptera, Cambodia, New record

We targeted three major Leptocorisa species (L. chinensis, L. acuta, and L. oratoria) and evaluated their potential distributions using MaxEnt. The results showed that most Asian countries and northern Australia would be suitable for at least one of these pest species, and climate change will expand their habitat northward. All of the developed models were evaluated to be excellent with AUC, TSS, and OR10%. Most of the recorded regions of the Leptocorisa species are consistent with the result of potential distributions predicted in this study. The results confirmed that the minimum temperature of the coldest month mainly influences the three Leptocorisa species distributions. The potential distributions of the three species cover major rice cultivation areas regardless of climate change, suggesting that it would be necessary to establish a sustainable control strategy for the pests.

Minipigs have remarkably similar physiology to humans, therefore, they it can be a good animal model for inflammation study. Thus, the conventional (serum chemistry, histopathology) and novel analytic tools [immune cell identification in tissue, cytokine level in peripheral blood mononuclear cells (PBMC) and serum, NF-kB target gene analysis in tissue] were applied to determine inflammation in Chicago Miniature Swine (CMS) minipig. Lipopolysaccharide (LPS)-induced acute systemic inflammation caused liver and kidney damage in serum chemistry and histopathology. Immunohistochemistry (IHC) also showed an increase of immune cell distribution in spleen and lung during inflammation. Moreover, NF-kB-target gene expression was upregulated in lung and kidney in acute inflammation and in heart, liver, and intestine in chronic inflammation. Cytokine mRNA was elevated in PBMC under acute inflammation along with elevated absolute cytokine levels in serum. Overall, LPS-mediated systemic inflammation affects the various organs, and can be detected by IHC of immune cells, gene analysis in PBMC, and measuring the absolute cytokine in serum along with conventional inflammation analytic tools.

Xenotransplantation has been considered an alternative to the moderate shortage of donor organs for transplantation. To achieve successful xenotransplatation, there is the need to overcome immune rejection. Although, hyperacute rejection has been overcome by α1,3-galactosyltransferase knockout pig, cellular immune rejection remains as a subsequent barrier. Interleukin-10 (IL-10) is known as an anti-inflammatory and immunomodulatory cytokine which has been shown to limit inflammatory responses by inhibiting macrophage activation in several animal experiments. To study the effect of human IL-10 (hIL-10) on pig-to-human xenotransplantation, porcine kidney epithelial cell line (PK(15)) expressing hIL-10 was established. The cytotoxicity of macrophages decreased by hIL-10 from transgenic cells. Furthermore, there is a decreased production of pro-inflammatory cytokines, tumor necrosis factor-α and interleukin-23, and increased anti-inflammatory cytokines like IL-10, but not transforming growth factor beta, in the presence of hIL-10. Also, macrophage polarization toward M2-like phenotype were induced by hIL-10 from transgenic PK(15) cells. Finally, we suggest that the cytotoxicity of human macrophages was reduced by hIL-10 from transgenic cells, inducing M2-like macrophage polarization. Therefore, these results show that hIL-10 transgenic pig can be used as a model to overcome acute immune rejection in pig-to-human xenotransplantation.

To investigate insect and plant community relationships in riparian zones, terrestrial insect communities were compared in plant communities in the riparian zone of the Miho River, Korea. The sweep netting method was used to sample insects in 50 m transects in three herbaceous plant communities. In 2020, each plant community—Chenopodium album, Beckmannia syzigachne, and Artemisia indica—was swept 100 times (50 sweeps × 2). In 2021, two communities had an additional 100 sweeps collected using 10 subsamples of 10 sweeps (excluding C. album communities). The surveyed dominant species or subdominant species of the insect community in each site preyed on the dominant plant species at the site. The Bray–Curtis similarity was significantly higher than the Sørensen similarity when comparing datasets across different years for the same plant species community. The predicted optimum sampling size to obtain approximately 80% of the total species estimated to be at each survey site, for effective quantitative collection of terrestrial insect herbivores in each plant community, was examined. Fifty sweeps were required for the A. indica community and 100 sweeps were required for the B. syzigachne community. The results of this study provide important data for riparian biodiversity conservation and future pest monitoring.

In vitro generation of dendritic cells (DCs) is advantageous for overcoming the low frequency of primary DCs and the difficulty of applying isolation techniques for studying DC immunobiology. The culture of bone marrow cells with granulocyte-macrophage colony-stimulating factor (GM-CSF) has been used extensively to generate bone marrow-derived dendritic cells (BMDCs). Studies have reported the heterogeneity of cells grown in murine GM-CSF culture based on the levels of MHCII expression. Although porcine DCs are generated by this classical method, the exact characteristics of the BMDC population have not yet been defined. In this study, we discriminated GM-CSF-grown BMDCs from gnotobiotic miniature pigs according to several criteria including morphology, phenotype, gene expression pattern and function. We showed that porcine BMDCs were heterogeneous cells that differentially expressed MHCII. MHCIIhigh cells displayed more representative of DC-like morphology and phenotype, including costimulatory molecules, as well as they showed a superior T cell priming capacity as compared to MHCIIlow cell. Our data showed that the difference in MHCIIhigh and MHCIIlow cell populations involved distinct maturation states rather than the presence of different cell types. Overall, characterization of porcine BMDC cultures provides important information about this widely used cellular model.

Sialyllactose (SL), an acidic oligosaccharide, has immune-protective effects against pathogens and helps with the development of the immune system and intestinal microorganisms. To elucidate the pharmacokinetic characterization after oral administration to rats, the simultaneous quantification method for 3′-SL and 6′-SL in rat plasma was validated, using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in an electrospray ionization (ESI) mode. Several types of columns [C18, amide, and hydrophilic interaction liquid chromatography (HILIC) phase] were used to separate the peaks of 3′-SL and 6′-SL, which improved chromatographic selectivity. Ultimately, the HILIC phase column had a good peak shape and quick resolution, with a mobile phase comprising ammonium acetate buffer and acetonitrile obtained by gradient elution. In addition, the simultaneous quantification of 3′-SL and 6′-SL in rat plasma samples were adequately applied to pharmacokinetic study.

Graphene, a two-dimensional carbon sheet with single-atom thickness, shows immense promise in several nanoscientific and nanotechnological applications, including in sensors, catalysis, and biomedicine. Although several studies have shown the cytotoxicity of graphene oxide in different cell types, there are no comprehensive studies on human embryonic kidney (HEK293) cells that include transcriptomic analysis and an in vitro investigation into the mechanisms of cytotoxicity following exposure to graphene oxide. Therefore, we exposed HEK293 cells to different concentrations of graphene oxide for 24 h and performed several cellular assays. Cell viability and proliferation assays revealed a significant dose-dependent cytotoxic effect on HEK293 cells. Cytotoxicity assays showed increased lactate dehydrogenase (LDH) leakage and reactive oxygen species (ROS) generation, and decreased levels of reduced glutathione (GSH) and increased level of oxidized glutathione indicative of oxidative stress. This detailed mechanistic approach showed that graphene oxide exposure elicits significant decreases in mitochondrial membrane potential and ATP synthesis, as well as in DNA damage and caspase 3 activity. Furthermore, our RNA-Seq analysis revealed that HEK293 cells exposed to graphene oxide significantly altered the expression of genes involved in multiple apoptosis-related biological pathways. Moreover, graphene oxide exposure perturbed the expression of key transcription factors, promoting these apoptosis-related pathways by regulating their downstream genes. Our analysis provides mechanistic insights into how exposure to graphene oxide induces changes in cellular responses and massive cell death in HEK293 cells. To our knowledge, this is the first study describing a combination of cellular responses and transcriptome in HEK293 cells exposed to graphene oxide nanoparticles, providing a foundation for understanding the molecular mechanisms of graphene oxide-induced cytotoxicity and for the development of new therapeutic strategies.

Ovarian cancer incidence continues to increase at an alarming rate. Although various therapeutic approaches exist for ovarian cancer, they have limitations, including undesired side effects. Therefore, nanoparticle (NP)-mediated therapy may be a viable, biocompatible, and suitable alternative. To the best of our knowledge, no comprehensive analysis has been undertaken on the cytotoxicity and cellular pathways involved in ovarian cancer cells, particularly SKOV3 cells. Here, we investigated the effect of palladium NPs (PdNPs) and the molecular mechanisms and cellular pathways involved in ovarian cancer. We assayed cell viability, proliferation, cytotoxicity, oxidative stress, DNA damage, and apoptosis and performed an RNA-Seq analysis. The results showed that PdNPs elicited concentration-dependent decreases in cell viability and proliferation and induced increasing cytotoxicity at increasing concentrations, as determined by leakage of lactate dehydrogenase, increased levels of reactive oxygen species and malondialdehyde, and decreased levels of antioxidants like glutathione and superoxide dismutase. Furthermore, our study revealed that PdNPs induce mitochondrial dysfunction by altering mitochondrial membrane potential, reducing adenosine triphosphate levels, inducing DNA damage, and activating caspase 3, all of which significantly induced apoptosis in SKOV3 cells following PdNPs treatment. Gene ontology (GO) term analysis of PdNPs-exposed SKOV3 cells showed various dysregulated pathways, particularly nucleosome assembly, telomere organization, and rDNA chromatin silencing. When genes downregulated by PdNPs were applied to GO term enrichment analysis, nucleosome assembly was the top-ranked biological pathway. We also provide evidence for an association between PdNPs exposure and multiple layers of epigenetic transcriptional control and establish a molecular basis for NP-mediated apoptosis. These findings provide a foundation, potential targets, and novel insights into the mechanism underlying toxicity and pathways in SKOV3 cells, and open new avenues to identify novel targets for ovarian cancer treatment.

Welding alloy 617 with other metals and alloys has been receiving significant attention in the last few years. It is considered to be the benchmark for the development of economical hybrid structures to be used in different engineering applications. The differences in the physical and metallurgical properties of dissimilar materials to be welded usually result in weaker structures. Fatigue failure is one of the most common failure modes of dissimilar material welded structures. In this study, fatigue life prediction of dissimilar material weld was evaluated by the accelerated life method and artificial neural network approach (ANN). The accelerated life testing approach was evaluated for different distributions. Weibull distribution was the most appropriate distribution that fits the fatigue data very well. Acceleration of fatigue life test data was attained with 95% reliability for Weibull distribution. The probability plot verified that accelerating variables at each level were appropriate. Experimental test data and predicted fatigue life were in good agreement with each other. Two training algorithms, Bayesian regularization (BR) and Levenberg⁻Marquardt (LM), were employed for training ANN. The Bayesian regularization training algorithm exhibited a better performance than the Levenberg⁻Marquardt algorithm. The results confirmed that the assessment methods are effective for lifetime prediction of dissimilar material welded joints.

In this study, dissimilar material welding between Alloy617 and 12Cr steel was performed using the buttering welding technique on the 12Cr steel side in order to increase the weldability. After multi-pass welding, post weld heat treatment (PWHT) was performed in order to reduce the welding residual stresses, and metallurgical microstructures were observed on dissimilar material weld. Additionally, the corrosion fatigue crack growth characteristics of dissimilar material weld was assessed according to Fracture Mechanics. Based on the results, the fatigue and corrosion fatigue strength of dissimilar material weld between Alloy617 and 12Cr steel did not show a big difference prior to and after post weld heat treatment. Fatigue and corrosion fatigue crack growth of dissimilar material weld were slightly faster than those of similar material weld of Alloy617. However, the characteristics of fatigue and corrosion fatigue crack growth did not show a big difference prior to and after PWHT.

The reliability of welded structure can be evaluated through welding residual stress analysis and fatigue strength assessment. In this study, welding residual stresses of multi-pass dissimilar material welded joint between alloy 617 and 12Cr steel were analyzed numerically and experimentally. Fatigue strength was then assessed in the air. Based on results of welding residual stress analysis and fatigue strength assessment, a fatigue design method considering welding residual stress was investigated. Welding residual stresses at the weld of dissimilar welded joints distributed complicatedly on longitudinal and transverse directions, showing differences but a very similar distribution tendency between numerical and experimental results. Numerical and experimental peak values of welding residual stresses at HAZ of the weld on the 12Cr steel side were predicted to be 333 MPa and 282 MPa HAZ, respectively. The fatigue limit of dissimilar material welded joint between alloy 617 and 12Cr steel was assessed to be 306.8 MPa, which was 40% of tensile strength (767 MPa) of dissimilar material welded joint. However, the stress range including welding residual stress was assessed to be 206.9 MPa, which was 14% lower than that calculated by including the effect of residual stresses.

The most effective method to reduce CO2 gas emission from the steam power plant is to improve its performance by elevating the steam temperature to more than 700 °C. For this, it is necessary to develop applicable materials at high temperatures. Ni-based Alloy 617 and 12Cr steel are used in steam power plants, due to their remarkable mechanical properties, high corrosion resistance, and creep strength. However, since Alloy 617 and 12Cr steel have different chemical compositions and thermal and mechanical properties, it is necessary to develop dissimilar material welding technologies. Moreover, in order to guarantee the reliability of dissimilar material welded structures, the assessment of mechanical and metallurgical properties, fatigue strength, fracture mechanical analysis, and welding residual stress analysis should be conducted on dissimilar material welded joints. In this study, first, multi-pass dissimilar material welding between Alloy 617 and 12Cr steel was performed under optimum welding conditions. Next, mechanical properties were assessed, including the static tensile strength, hardness distribution, and microstructural analysis of a dissimilar material welded joint. The results indicated that the yield strength and tensile strength of the dissimilar metal welded joint were higher than those of the Alloy 617 base metal, and lower than those of the 12Cr steel base metal. The hardness distribution of the 12Cr steel side was higher than that of Alloy 617 and the dissimilar material weld metal zone. It was observed that the microstructure of Alloy 617 HAZ was irregular austenite grain, while that of 12Cr steel HAZ was collapsed martensite grain, due to repeatable heat input during multi-pass welding.

Introduction: Decreasing rates of blood donation and close margins between blood supply and demand pose challenges in healthcare. Genetically engineered pig red blood cells (pRBCs) have been explored as alternatives to human RBCs for transfusion, and triple-gene knockout (TKO) modification improves the compatibility of pRBCs with human blood in vitro. In this study, we assessed the efficacy and risks of transfusing wild-type (WT)- and TKO-pRBCs into nonhuman primates (NHPs). Methods: Blood from O-type WT and TKO pigs was processed to produce pRBCs for transfusion, which were transfused or not into NHPs (n=4 per group: WT, TKO, and control) after 25% total blood volume withdrawal: their biological responses were compared. Hematological, biochemical, and immunological parameters were measured before, immediately after, and at intervals following transfusion. Two months later, a second transfusion was performed in three NHPs of the transfusion group. Results: Transfusion of both WT- and TKO-pRBCs significantly improved RBC counts, hematocrit, and hemoglobin levels up to the first day post-transfusion, compared to the controls. The transfusion groups showed instant complement activation and rapid elicitation of anti-pig antibodies, as well as elevated liver enzyme and bilirubin levels post-transfusion. Despite the higher agglutination titers with WT-pRBCs in the pre-transfusion crossmatch, the differences between the WT and TKO groups were not remarkable except for less impairment of liver function in the TKO group. After the second transfusion, more pronounced adverse responses without any hematological gain were observed. Conclusions: WT- and TKO-pRBC transfusions effectively increased hematologic parameters on the first day, with rapid clearance from circulation thereafter. However, pRBC transfusion triggers strong antibody responses, limiting the benefits of the pRBC transfusion and increasing the risk of adverse reactions. [ABSTRACT FROM AUTHOR]