• Journal of Animal Science and Technology
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• v.64 no.5
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• pp.842-853
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• 2022

Ochratoxin A (OTA) is a well-known mycotoxin that causes disease through the ingestion of contaminated food or feed, for example, in the porcine industry. The intestinal epithelium acts as the first barrier against food contamination. We conducted a study on the exposure of the porcine intestinal epithelium to OTA. We used the intestinal porcine epithelial cell line IPEC-J2 as an in vitro model to evaluate the altered molecular mechanisms following OTA exposure. Gene expression profiling revealed that OTA upregulated 782 genes and downregulated 896, totalling 1678 differentially expressed genes. Furthermore, immunofluorescence, quantitative real-time polymerase chain reaction, and western blotting confirmed that OTA damages the tight junction protein ZO-1. Moreover, OTA activated the expression of inflammatory genes (IL-6, IL-8, IL-10, NF-kB, TLR4, and TNF-α). In summary, this study confirmed that OTA alters various molecular mechanisms and has several adverse effects on IPEC-J2 cells.

• Journal of Life Science
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• v.34 no.9
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• pp.666-672
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• 2024

Plants recognize pathogens through intracellular receptors that trigger defense signaling. Nucleotide-binding leucine-rich repeat (NLR) proteins within a cell specifically recognize pathogenic molecules (effectors), leading to signal transduction that ultimately triggers the cell death pathway, thereby inducing effector-triggered immunity in plants. NLR proteins are broadly categorized into two types based on their N-terminal domains: coiled-coil domain NLRs (CNLs) and toll/interleukin-1 receptor (TIR) domain NLRs (TNLs) are defined by their unique N-terminal domains. The TIR domain, which is responsible for activates nicotinamide adenine dinucleoside hydrolases (NADases), is crucial for the degradation of the NAD+ cofactor. TNL-dependent immune signaling involves lipase-like proteins known as Enhanced Disease Susceptibility 1 (EDS1) and its partners Phytoalexin Deficient 4 (PAD4) and Senescence-Associated Gene 101 (SAG101). This immune system also requires helper NLR subfamilies, such as activated disease resistance 1 (ADR1) and N requirement gene 1 (NRG1). The catalytic activity of TIR domain proteins generates various small molecules reported to activate plant's immune responses. These small molecules bind to specific sites on EDS1-PAD4 and EDS1-SAG101, inducing structural changes in the EP domain, and subsequently enabling interaction with ADR1 or NRG1. Here, we will discuss the characteristics of these small molecules and describe their relationships with protein complexes based on their structural and biochemical characteristics. We will also discuss how these small molecules can activate immune pathways.

• Journal of Chest Surgery
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• v.31 no.12
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• pp.1200-1205
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• 1998

Background: Lung cancer formation is a multistage process involving activation of protooncogene and inactivation of tumor suppressor genes. We evaluate the significance of cyclin D1, p53, bcl-2 gene mutations in patients with curatively resected stage IIIA non-small cell lung cancer(NSCLC). Material and Method: One hundred consecutive cases of stage IIIA lung cancers from patients operated on curatvely between 1990 and 1995 for which adequate paraffin blocks and clinical history were available. Immunohistochemical studies were performed on the representative tissue sections from each case by the labelled streptovidin- biotin method. Sections for cyclin D1, p53, Bcl-2 immunostaining were pretreated in a microwave oven for 10 to 20 minutes in citrate buffer before immunostaining. The overnight incubation with NCL-cyclin D1-GM for cyclin D1, with clone DO-7 for p53, with clone 124 for bcl-2 was done. Mean follow-up was 24.1 months (range 2-84 months) after operation. Result: One hundred cases of lung cancers were composed of 56 cases of squamous cell carcinoma, 37 cases of adenocarcinoma, 5 cases of adenosquamous cell carcinoma, and 2 cases of large cell carcinoma. The 5-year survival was 32.1%. The positive expression rate of cyclin D1 was 35%, p53 was 56%, and bcl-2 was 17%. But there were no correlation between cyclin D1, p53, Bcl-2 protein expression and survival. Conclusion: These observation indicate that cyclin D1, p53, bcl-2 protein overexpression might be implicated in the oncogenesis of non-small cell lung carcinomas but they have no usefulness as a prognostic marker.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.8
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• pp.3301-3306
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• 2015

Nucleolin (C23) is an important anti-apoptotic protein that is ubiquitously expressed in exponentially growing eukaryotic cells. In order to understand the impact of C23 in radiation therapy, we attempted to investigate the relationship of C23 expression with the radiosensitivity of human non-small cell lung cancer (NSCLC) cells. We investigated the role of C23 in activating the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs), which is a critical protein for DNA double-strand breaks (DSBs) repair. As a result, we found that the expression of C23 was negatively correlated with the radiosensitivity of NSCLC cell lines. In vitro clonogenic survival assays revealed that C23 knockdown increased the radiosensitivity of a human lung adenocarcinoma cell line, potentially through the promotion of radiation-induced apoptosis and adjusting the cell cycle to a more radiosensitive stage. Immunofluorescence data revealed an increasing quantity of ${gamma}$-H2AX foci and decreasing radiation-induced DNA damage repair following knockdown of C23. To further clarify the mechanism of C23 in DNA DSBs repair, we detected the expression of DNA-PKcs and C23 proteins in NSCLC cell lines. C23 might participate in DNA DSBs repair for the reason that the expression of DNA-PKcs decreased at 30, 60, 120 and 360 minutes after irradiation in C23 knockdown cells. Especially, the activity of DNA-PKcs phosphorylation sites at the S2056 and T2609 was significantly suppressed. Therefore we concluded that C23 knockdown can inhibit DNA-PKcs phosphorylation activity at the S2056 and T2609 sites, thus reducing the radiation damage repair and increasing the radiosensitivity of NSCLC cells. Taken together, the inhibition of C23 expression was shown to increase the radiosensitivity of NSCLC cells, as implied by the relevance to the notably decreased DNA-PKcs phosphorylation activity at the S2056 and T2609 clusters. Further research on targeted C23 treatment may promote effectiveness of radiotherapy and provide new targets for NSCLC patients.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.7
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• pp.949-955
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• 2019

Objective: The present study was to investigate the association of polymorphisms in exon-9 of the bone morphogenetic protein receptor-1B (BMPR-1B) gene (C864T) with litter size in 240 Dorset, 232 Mongolian, and 124 Small Tail Han ewes. Methods: Blood samples were collected from 596 ewes and genomic DNA was extracted using the phenol: chloroform extraction method. The 304-bp amplified polymerase chain reaction product was analyzed for polymorphism by single-strand conformation polymorphism method. The genotypic frequency and allele frequency of BMPR-1B gene exon-9 were computed after sequence alignment. The ${\chi}^2$ independence test was used to analyze the association of genotypic frequency and litter size traits with in each ewe breed, where the phenotype was directly treated as category. Results: The results indicated two different banding patterns AA and AB for this fragment, with the most frequent genotype and allele of AA and A. Calculated Chi-square test for BMPR-1B gene exon-9 was found to be more than that of p value at the 5% level of significance, indicating that the population under study was in Hardy-Weinberg equilibrium for all ewes. The ${\chi}^2$ independence test analyses indicated litter size differences between genotypes was not the same for each breed. The 304-bp nucleotide sequence was subjected to BLAST analysis, and the C864T mutation significantly affected litter size in singletons, twins and multiples. The heterozygosity in exon-9 of BMPR-1B gene could increase litter size for all the studied ewes. Conclusion: Consequently, it appears that the polymorphism BMPR-1B gene exon-9 detected in this study may have potential use in marker assisted selection for litter size in Dorset, Mongolian, and Small Tail Han ewes.

• Korean Journal of Clinical Laboratory Science
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• v.56 no.3
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• pp.198-206
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• 2024

EZH2 is a methyltransferase that is a critical target for lymphoma treatment. However, it is not yet widely used in clinical settings. PROteolysis TArgeting Chimeras (PROTACs) represent a novel therapeutic strategy aimed at eliminating proteins that have been a challenging target using conventional small molecules. In our previous research, we compared the small molecules-based EZH2 inhibitor used in clinical settings with a PROTAC-based EZH2 degrader. We found that the PROTAC-based degrader was significantly more effective. Building on this, we further investigated the effects of combining the PROTAC-based EZH2 degrader (dEZH2) with a METTL3 inhibitor, both of which have demonstrated effectiveness in inhibiting cell proliferation and inducing apoptosis in Burkitt's lymphoma. Using the CCK-8 assay, we found that both drugs, alone and in combination, significantly inhibited Daudi and Ramos cell growth in a dose-dependent manner. The combined treatment markedly suppressed cell proliferation and induced apoptosis, as confirmed by Annexin V/PI staining. Our results revealed G2/M phase arrest with a significant decrease in the G0/G1 phase by flow cytometry. Our study also showed increased levels of cleaved PARP, cleaved caspase-3, tumor protein p53 (TP53), and PUMA using the western blot technique, indicating enhanced p53-dependent apoptosis. Our findings suggest that the combination therapy of dEZH2 and iMETTL3 could be a promising approach in the treatment of Burkitt's lymphoma.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.5
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• pp.659-666
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• 2009

Metabolizable protein (MP) supply and amino acid balance in the intestine were manipulated through selection of highly digestible rumen-undegradable protein (RUP) sources and protected methionine (Met) supplementation. Four ruminallycannulated, multiparous Holstein cows averaging 193${\pm}$13 days in milk were used in a 4${\times}$4 Latin square design to assess N utilization and milk production responses to changes in RUP level, post-ruminal RUP digestibility and protected Met supplementation. Treatments were A) 14.0% crude protein (CP), 8.0% rumen degradable protein (RDP) and 6.0% RUP of low intestinal digestibility (HiRUP-LoDRUP); B) 14.1% CP, 8.1% RDP and 6.0% RUP of high intestinal digestibility (HiRUP-HiDRUP); C) 13.1% CP, 7.9% RDP and 5.2% RUP of high intestinal digestibility (LoRUP-HiDRUP), and D) 13.1% CP, 7.9% RDP and 5.2% RUP of high intestinal digestibility plus rumen escape sources of Met (LoRUP-HiDRUP+Met). Experimental diets were formulated to have similar concentrations of RDP, net energy of lactation ($NE_L$), neutral detergent fiber (NDF), acid detergent fiber (ADF), calcium, phosphorus and ether extract using the NRC model (2001). Results showed that dry matter intake (DMI), production of milk fat and protein were similar among treatments. Milk production was similar for diet HiRUP-LoDRUP, HiRUP-HiDRUP and LoRUP-HiDRUP+Met, and significantly higher than diet LoRUP-HiDRUP. Milk fat and protein percentage were higher for cows receiving HiDRUP treatments, with the greatest increases in the diet LoRUP-HiDRUP+Met. There was no significant change in ruminal pH, $NH_3g-N$ and volatile fatty acid (VFA) concentration among all treatments. Apparent digestibility of dry matter (DM), CP, NDF and ADF and estimated bacterial CP synthesis were similar for all treatments. Nitrogen intakes, blood and milk urea-N concentrations were significantly higher for cows receiving HiRUP diets. Urine volume and total urinary N excretion were significantly lowered by LoRUP diets. Lowering dietary RUP level while supplementing the highly digestible RUP source with rumen escape sources of Met resulted in similar milk production, maximal milk fat and protein concentration and maximum N efficiency, indicating that post-ruminal digestibility of RUP and amino acid balance in the small intestine can be more important than total RUP supplementation.

• Korean Journal of Food Science and Technology
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• v.20 no.6
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• pp.883-889
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• 1988

The salted small shrimps(Acetes japonicus) were fermented for 3 months at room temperature. During the period of fermentation, the changes of shrimp protein properties were determined. The extractability of soluble protein was slightly decreased in 1 month fermentation, but thereafter increased. The contents of 10% TCA soluble fraction were gradually increased during 3 month fermentation, and the rate of 10% TCA soluble fraction/total soluble protein was also greatly increased during the period of fermentation. Sephadex G-100 gel filtration pattern was changed after 1 month fermentation, showing the disappearance of low molecular weight protein peaks, the decomposition and the delay of elution time of main shrimp protein peaks. Polyacrylamide gel disc electrophoresis patterns showed the degradation of main protein bands into lots of smaller bands after 1 month fermentation. The contents of total free amino acids were slightly decreased in 1 month fermentation and then gradually increased during the Period of fermentation. The rate of free amino acids/soluble protein was steadily increased during the period of fermentation, but the rate of free amino acids/10% TCA soluble fraction was decreased continually during the period of fermentation. The contents of most free amino acids were increased during the period of fermentation, but those of histidine and arginine were greatly decreased in 1 month fermentation. Ammonia was increased after 1 month fermentation. The pH value of salted shrimp was slowly changed during 3 months of fermentation, showing increase from 7.8 to 8.2.

• Korean Journal of Poultry Science
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• v.12 no.1
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• pp.39-44
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• 1985

These studies were conducted to compare the nutritive values and optimum prices of eggs among 6 groups of different egg Weight. With the total of 100 eggs of each weight group, after the weight percentage of egg yolk, albumen and shell in the whole egg were investigated, protein and fat contents of e99 yolk and albumen were analyzed. and then protein and fat contents in the whole eggs were calculated. Finally, the optimum prices of eggs in relation to the egg weight were studied on the basis of egg weight, protein content and protein plus fat contents of eggs, respectively. The results obtained are summarized as follows; 1. As the egg weight (X, g/10 eggs) increased, egg yolk (Y$_1$, %) and shell(Y$_2$, %) percentages tended to decrease, but egg albumen(Y$_3$, %) percentage increased lineally; Y$_1$=44.34-0.02X, Y$_2$=15.358-0.006 X, and Y$_3$=40.136＋0.026 X. 2. There were no significant differences in protein and fat contents of eggs among 6 different groups of egg weight. 3. Protein (Y$_1$, %), fat (Y$_2$, %) and protein plus fat (Y$_3$, %) contents in the whole eggs declined progressively as the egg weight (X, g/10 eggs) increased ; Y$_1$=11.943-0.00032X, Y$_2$=13.996-0.00614X, and Y$_3$=25.939-0.00646X. 4. Similar results were obtained whether the optimum prices of eggs were estimated on the basis of egg weight or protein content of eggs, and they were higher in the large size eggs and lower in the small size eggs than the optimum prices of eggs estimated on the basis of protein plus fat content of eggs.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.2
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• pp.205-212
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• 2010

The effects of chromium (Cr), dietary crude protein (CP) level, and potential interactions of these two factors were investigated in term of energy metabolism in lambs. Forty-eight 9-week-old weaned lambs (Dorper${\times}$Small-tail Han sheep, male, mean initial body weight = 22.96 kg${\pm}$2.60 kg) were used in a 2${\times}$3 factorial arrangement of supplemental Cr (0 ${\mu}g$/kg, 400 $\mu{g}$/kg or 800 ${\mu}g$/kg from chromium yeast) and protein levels (low protein: 157 g/d to 171 g/d for each animal, or high protein: 189 g/d to 209 g/d for each animal). Blood samples were collected at the beginning and end of the feeding trial. The lambs were then sacrificed and tissue samples were frozen for further analysis. Chromium at 400 ${\mu}g$/kg decreased fasting insulin level and the ratio of plasma insulin to glucagon, but these differences were not statistically significant; in contrast, chromium at 800 ${\mu}g$/kg increased the ratio significantly (p<0.05). Protein at the high level increased plasma tumor necrosis factor $\alpha$ (TNF-$\alpha$) level (p = 0.060). Liver glycogen content was increased significantly by Cr (p<0.05), which also increased liver glucose-6-phosphatase (G-6-Pase) and adipose hormone-sensitive lipase (HSL) activity. At 400 ${\mu}g$/kg, Cr increased muscle hexokinase (HK) activity. High protein significantly increased G-6-Pase activities in both the liver (p<0.05) and the kidney (p<0.05), but significantly decreased fatty acid synthase (FAS) activity in subcutaneous adipose tissue (p<0.05). For HSL activity in adipose tissue, a Cr${\times}$CP interaction (p<0.05) was observed. Overall, Cr improved energy metabolism, primarily by promoting the glycolytic rate and lipolytic processes, and these regulations were implemented mainly through the modulation by Cr of the insulin signal transduction system. High protein improved gluconeogenesis in both liver and kidney. The interaction of Cr${\times}$CP indicated that 400 $\mu{g}$/kg Cr could reduce energy consumption in situations where energy was being conserved, but could improve energy utilization when metabolic rate was increased.