METTL17 coordinates ferroptosis and tumorigenesis by regulating mitochondrial translation in colorectal cancer.

Ferroptosis, an iron-dependent lipid peroxidation-induced form of regulated cell death, shows great promise as a cancer therapy strategy. Despite the critical role of mitochondria in ferroptosis regulation, the underlying mechanisms remain elusive. This study reveals that the mitochondrial protein METTL17 governs mitochondrial function in colorectal cancer (CRC) cells through epigenetic modulation. Bioinformatic analysis establishes that METTL17 expression positively correlates with ferroptosis resistance in cancer cells and is up-regulated in CRC. Depletion of METTL17 sensitizes CRC cells to ferroptosis, impairs cell proliferation, migration, invasion, xenograft tumor growth, and AOM/DSS-induced CRC tumorigenesis. Furthermore, suppression of METTL17 disrupts mitochondrial function, energy metabolism, and enhances intracellular and mitochondrial lipid peroxidation and ROS levels during ferroptotic stress. Mechanistically, METTL17 inhibition significantly reduces mitochondrial RNA methylation, including m4C, m5C, m3C, m7G, and m6A, leading to impaired translation of mitochondrial protein-coding genes. Additionally, the interacting proteins associated with METTL17 are essential for mitochondrial gene expression, and their knockdown sensitizes CRC cells to ferroptosis and inhibits cell proliferation. Notably, combined targeting of METTL17 and ferroptosis in a therapeutic approach effectively suppresses CRC xenograft growth in vivo. This study uncovers the METTL17-mediated defense mechanism for cell survival and ferroptosis in mitochondria, highlighting METTL17 as a potential therapeutic target for CRC.

Emergence of multidrug-resistant Bacillus spp. derived from animal feed, food and human diarrhea in South-Eastern Bangladesh.

BACKGROUND: Antimicrobial resistance poses a huge risk to human health worldwide, while Bangladesh is confronting the most severe challenge between the food supply and the huge consumption of antibiotics annually. More importantly, probiotics containing Bacillus spp. are claimed to be an alternative to antimicrobial stewardship programs. However, their antibiotic resistance remains elusive. Thus, we employed the antimicrobial susceptibility test and PCR to assess the prevalence of resistance, including multidrug resistance (MDR) and resito-genotyping of isolated Bacillus spp. RESULTS: The phenotypic profile showed that Bacillus spp. were 100% sensitive to gentamicin (2 µg/mL), whereas lowered sensitivity to levofloxacin (67.8%, 0.5-1 µg/mL), ciprofloxacin (62.3%, 0.5-1 µg/mL), clindamycin (52.2%, 0.25-0.5 µg/mL), amoxicillin-clavulanic acid (37.6%, 0.06 µg/mL), azithromycin (33.4%, 1-2 µg/mL), tetracycline (25.6%, 2-4 µg/mL), nitrofurantoin (21.1%, 16-32 µg/mL), co-trimoxazole (19.2%, 2 µg/mL), and erythromycin (18.8%, 0.25-0.5 µg/mL). The strains were completely resistant to penicillin, amoxicillin-clavulanic acid, cefixime, ceftriaxone, vancomycin, and co-trimoxazole, and a species-specific trend was seen in both phenotypic and genotypic resistance patterns. Genotypic resistance indicated prevalence of the bla1 (71.5%), tetA (33%), erm1 (27%), blaTEM (13.1%), blaCTX-M-1/blaCTX-M-2 /sul1 (10.1%), blaSHV (9.6%), and qnrS (4.1%) genes. The ß-lactamase resistance gene bla1 was found in all penicillin-resistant (MIC ≥ 32 µg/mL) Bacillus spp. One hundred ninety-one isolates (89.6%) were MDR, with 100% from diarrhea, 90.3% from food, and 88.7% from animal feed. CONCLUSION: Based on the MIC value and profile analysis of antibiotic resistance genes, this is the first study that Bacillus spp. antimicrobial susceptibilities have been identified in Bangladesh, and our study will shed light on the adverse effects of feed-borne Bacillus spp. emerging from animal feed to the food chain. A comprehensive investigation is urgently needed by policymakers on tolerance limits and harmful effects in the animal industry.

Physcion, a novel anthraquinone derivative against Chlamydia psittaci infection.

Physcion, a natural anthraquinone derivative, has been reported to exert remarkable antibacterial activities against Staphylococcus aureus,Staphylococcus epidermidis and Pseudomonas aeruginosa. However, it is not fully illustrated as anti-Chlamydia substance. In the present study, minimum inhibitory concentration(MIC)values for physcion against Chlamydia psittaci(C.psittaci) 6BC, C.psittaci SBL and C.psittaci HJ were 128 µg/mL,256 µg/mL and 128 µg/mL while minimum bactericidal concentration (MBC) values were 256 µg/mL,512 µg/mL and 256 µg/mL,respectively. Moreover, Chlamydial adhesion to Hela 229 cells was blocked in a dose-dependent manner and RB-to-EB differentiation was inhibited by physcion from 28 to 48 hpi.Post treatment,upregulation of LC3-II was in a dose-dependent manner, indicating physcion activated autophagy and bacterial clearance.To validate clinical efficacy,49 SPF chickens aged 21days were divided into 5 groups and infected intra-laryngeally with 0.2 mL of 1 × 107 IFU/mL C.psittaci 6 BCE.Three days later, birds received orally with serial doses of physcion (4 mg/kg to 9 mg/kg), or 3 mg/kg of doxycycline for 6 days.Chickens with difficulty in breathing were alleviated significantly with increasing concentrations of physicon.Postmortem,lesions of air sacs were reduced significantly in a dose-dependent manner.More importantly,birds with 9 mg/kg of physcion could alleviate lesions of air sacs and lungs, and reduce bacterial loads in spleens, which was comparable to doxycycline treatment. Based on above evidences, physcion is a promising cost-effective natural drug by blocking Chlamydial adhesions to host cells, RB-to-EB differentiation and activating bacterial autophagy and it will be a good alternative to doxycycline combating virulent C.psittaci infection, contributing to eradication of Chlamydial transmission from animals to human beings.

Protocatechuic acid: A novel detoxication agent of fumonisin B1 for poultry industry.

Fumonisin B1 (FB1) is a major fusarium mycotoxin that largely contaminates feedstuffs and foods, posing a health risk to both animals and humans. This mycotoxin can enter the human body directly through contaminated food consumption or indirectly by toxins and their metabolites. In a prior study, feed-borne FB1 is one of the leading mycotoxins in breeder eggs, leading to reduced hatchability and gizzard ulceration in chicken progenies. Currently, no effective way is available to remove FB1 from feeds and human-contaminated foods. We hypothesize that FB1 can be reduced to low risk by protocatechuic acid (PCA). To assess the ability of FB1 to be degraded in vivo, 1 ppm of FB1 was treated with PCA, or D-glucose, or silymarin, or anti-FB1 monoclonal antibody. Our study revealed that both D-glucose and PCA exhibited 53.4 and 71.43% degradation, respectively, at 80°C for 2 h, while 35.15% of FB1 detoxification was determined in the silymarin group at 60°C for 0.5 h. A dose-dependent manner was found after treatment with D-glucose or PCA at 80°C for 2 h. As for detoxification of anti-FB1 monoclonal antibody, the 1:3,000 dilution induced significant FB1 detoxification, accounting for 25.9% degradation at 25°C for 2 h. Furthermore, 50 SPF 11-day-old embryonated eggs were divided into 10 groups, with five eggs per group. Post treatment with PCA or D-glucose, or silymarin or anti-FB1 monoclonal antibody, the treated samples were inoculated into albumens and monitored daily until the hatching day. Consequently, 100% of the chickens survived in the D-glucose group and other control groups, except for the FB1 control group, while 80, 80, and 60% hatching rates were found in the PCA-treated group, the anti-FB1 monoclonal antibody-treated group, and the silymarin-treated group. Additionally, both the FB1 group and the silymarin-treated group yielded lower embryo growth than other groups did. Postmortem, lower gizzard ulceration index was determined in the PCA-treated group and the anti-FB1 monoclonal antibody-treated group compared to those of the silymarin-treated group and D-glucose-treated group. Based on the above evidence, PCA is a promising detoxification to reduce FB1 contamination in the poultry industry, contributing to the eradication of mycotoxin residuals in the food chain and maintaining food security for human beings.

Infectious bursal disease virus replication is inhibited by avain T cell chemoattractant chemokine CCL19.

Chemokine CCL19, together with its receptor CCR7, is one of the most important factors recruiting immune cells into target organ during virus infection. Our previous study has shown that CCL19 played a vital role in the process of T cell trafficking into bursae during bursal disease virus (IBDV) infection. In this study, we hypothesized that CCL19 could exert direct influences on IBDV replication other than recruiting immune cells. A eukaryotic expression vector of pEGFP-N1/CCL19 was successfully constructed and identified by PCR, double enzymes digestion, and sequencing. Different concentrations of pEGFP-N1/CCL19 plasmids were transfected into DF1 cells and CCL19 protein was highly expressed. Then, DF1 cells were infected with IBDV B87 strain post-transfection. Based on PCR and Western blot results, CCL19 could obviously decrease the gene levels of VP1 and VP2 and the protein levels of VP2 and VP3. When CCL19 was knocked down, the gene levels of VP1 and VP2 were significantly upregulated. Moreover, indirect immunostaining revealed that the IBDV content was largely decreased after CCL19 overexpression. Additionally, CCL19 inhibitory effects might rely on activation of the JNK signal pathway. Taken together, chemokine CCL19 directly blocks IBDV replication in DF1 cells, indicating that CCL19 could play crucial functions other than recruiting T cells during the pathogenesis of IBDV.

Identification and assessment of pathogenicity of a naturally reassorted infectious bursal disease virus from Henan, China.

Infectious bursal disease virus (IBDV) is still a vital etiological agent in poultry farms. IBDV outbreaks occasionally occur due to the presence of very virulent, reassortment or variant strains. Vaccine immunization has played crucial roles in IBD control for decades. However, survival pressure of IBDV from the vaccine immunization also increases the reassortments of circulating viruses. In this study, an IBDV strain was isolated from several broiler farms in Henan Province, central part of China, and named IBDV HN strain. Based on the results of RT-PCR, sequencing and phylogenic analyses of VP1 and VP2 genes, the IBDV HN strain is a novel reassortment strain in the Henan region. Segment A of this strain appears to originate from the very virulent IBDV strain, while segment B comes from the other field reassortment strains. This may be the result of natural reassortant of virus circulating in the field. About 60% (6/10) of experimentally infected specific pathogen-free chickens died after 3 to 5 d post-infection with typical symptom and pathological lesions. The IBDV HN strain was prone to horizontal transmission, which poses a serious threat to the chicken industry. Further investigation on the prevalence, virulence, and evolution of HN strain IBDV will provide a foundation for the prevention and control of the disease in this region.

High-efficiency production of bioactive oleosin-basic fibroblast growth factor in A. thaliana and evaluation of wound healing.

Basic fibroblast growth factor (bFGF) is a member of the fibroblast growth factors family. It is a highly specific mitogenic factor for many cell types, as though it be involved in wound repair, angiogenesis, nerve nutrition and embryonic development etc. Oil bodies have been applied for medicine, foodstuff and industry field. The heterogonous proteins expressed in oil bodies have distinct advantages, such as less purification steps and low costs. In this study, bFGF was expressed in A. thaliana seeds using oleosin fusion technology. The pOTB-bFGF vector contained an oleosin-bFGF fusion gene and a glufosinate resistance gene for selection. Transgenic A. thaliana lines were obtained by the floral dip method and protein expression was identified by SDS-PAGE and western blotting in transgenic A. thaliana lines. Moreover, MTT assays showed that the oil bodies expressed oleosin-bFGF fusion protein had a remarkable proliferation effect on NIH/3T3 cells and animal experiments showed that it could effectively decrease wound size and accelerate granulation tissue maturation. In conclusion, this may be a better method of producing oleosin-bFGF fusion protein to meet the increasing demand in its pharmacological application.