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s For investigating the possible effect of Lactobacillus casei on the immune status of Shabot, Tor grypus , 480 fish weighing 40 ± 10 g were randomly divided into four groups in triplicate. The first group (control) was fed on a commercial diet without probiotic supplementation. Other groups (A, B, and C) were received diet supplemented by different levels of L. casei , i.e., 5 × 10 6 , 5 × 10 7 , and 5 × 10 8  CFU g −1 , respectively. The feeding trial was performed for 60 continuous days followed by 15 days as withdraw period, i.e., day 75. Serum immune responses were determined on days 0, 30, 60, and 75 of feeding trial. Gene expression in the kidneys was analyzed on days 0, 60, and 75 as well as following bacterial challenge. The results showed that dietary L. casei supplementation significantly ( P  < 0.05) increased serum lysozyme, serum bactericidal, complement, and respiratory burst activities as compared with control group, while it was observed that treated fish did not show any significant differences in immune responses on day 75 ( P  < 0.05). Results of gene expression (TNF-α, IL-1β, and IL-8) showed significant increases among different experimental times ( P  < 0.05). According to obtained results, it might be concluded that the feeding by L. casei could likely enhance the immune responses and gene expression in T. grypus . Treatments A and C were better than B, but the lower concentration is better as an economic viewpoints and biosecurity. Therefore, concentration A is suggested.

The prevalence of pulmonary disease caused by nontuberculous mycobacteria (NTM) is reportedly on the rise in the world. Some of the species are resistant to various antibiotics; hence, limited treatment options are available. The aims of this study were to investigate the prevalence of NTM and to determine the effect of d-cycloserine against and isolated from clinical specimens to find out the synergistic effect of d-cycloserine and clarithromycin. A total of 95 nonduplicate pulmonary isolates of NTM were collected from three major Regional Tuberculosis (TB) Centers. NTM isolates were identified by conventional tests and PCR sequence analysis of the gene. PCR sequencing of was performed for detecting the inducible resistance to macrolides. In vitro susceptibilities and activities of d-cycloserine-clarithromycin combinations were accessed using the broth microdilution method. Among 714-positive acid-fast bacilli from TB-suspected cases, 95 isolates were identified as NTM (13.3%). The prevalence of identified isolates was as follows: 46 (48.4%), 16 (16.8%), 15 (15.7%), 7 (7.3%), 4 (4.2%), 3 (3.2%), 2 (2.1%), and 2 (2.1%). In addition, sequence analysis could identify all NTM isolates. The effect of d-cycloserine was better than that of clarithromycin. The synergistic effect of d-cycloserine with clarithromycin was observed for six (100%) and five (71.5%) strains of and , respectively. In the present study, we demonstrated a wide range of NTM in processed samples from different provinces of Iran. Our observations indicated that d-cycloserine was very active against and ; hence, d-cycloserine, either alone or in combination with clarithromycin, may be promising for the treatment of - and associated diseases.

The purpose of this study was to investigate the function of the NLRP3-Caspase 1 signaling pathway in the colon during fecal microbiota transplantation (FMT) in colitis induced by acetic acid. Additionally, the study aimed to determine the impact of FMT on anxiety behaviors by analyzing the function of the NLRP3-Caspase 1 signaling pathway in the hippocampus. A total of twenty-four rats were selected randomly for the study and divided into two groups, a control group, and an acid acetic-induced colitis group. The acid acetic-induced colitis group further consisted of three subgroups: untreated acid acetic-induced colitis group, mesalazine 0.3 gr/kg group, and FMT group. After 6 days, the colon was evaluated for macroscopic and microscopic damage, and the signaling pathway NLRP3-Caspase1-related genes in the colon and hippocampus were analyzed. Additionally, anxiety-related behaviors of the rats were observed. FMT decreased colonic mRNA expression levels of NLRP3, NF-кB, and Caspase1 and pro-inflammatory cytokines (IL-1β and IL-18). Also, FMT reduced the expression of NLRP3, NF-κB, and Caspase1 protein levels as well as pro-inflammatory cytokines IL-1β and IL-18 in the hippocampus, resulting in a reduction of anxiety behaviors in the open field and elevated plus maze tests in the colitis model. FMT may improve acetic acid-induced colitis by regulating the NLRP3-Caspase1 signaling pathway in the colon. It also reduced colitis-induced anxiety behavior by regulating the expression of proteins related to the NLRP3-Caspase 1 pathway in the hippocampus.

Gastric ulcers are a type of digestive disease that can severely affect a person's quality of life. Our study aimed to investigate the effects of fish oil on ethanol-induced gastric ulcers in rats, with the purpose of providing more comprehensive information on the topic. The study looked at various factors such as gastric ulcer index, and nitric oxide (NO) levels in stomach tissue. To investigate apoptosis, the mRNA levels of Bax, Bcl-2, and Caspase 3 were analyzed. The results showed that fish oil can reduce gastric acidity and the gastric ulcer index in cases of ethanol-induced gastric ulcers. It was found that fish oil can increase NO levels and improve the anti-apoptotic system by increasing the expression of Bcl-2 while decreasing the expression of Bax and Caspase 3. In general, the study demonstrates that fish oil can protect the stomach from ethanol-induced damage by reducing the apoptosis pathway via nitric oxide.

Neonatal hypoxia has a negative impact on the developing brain during the sensitive period. Inflammation plays a key role in the physiological response to hypoxic stress. Considering the anti-inflammatory properties of alpha-pinene, which has received a lot of attention in recent years, in this research we focused on the impact of alpha-pinene on the behavioral responses and proinflammatory factors in rats subjected to the neonatal hypoxia. This study involved Wistar rats (7-day-old) that were divided into six experimental groups, including a control group, groups receiving different doses of alpha-pinene (5 and 10 mg/kg), a hypoxia group receiving 7% O 2 and 93% N 2 , 90 min duration for 7 days, and groups receiving alpha-pinene 30 min before hypoxia. All injections were done intraperitoneally. The rats were evaluated for proinflammatory factors 24 h after exposure to hypoxia (PND14) and at the end of the behavioral test (PND54). The results showed that hypoxia led to decreased motor activity, coordination, and memory, as well as increased inflammation. However, the rats that received alpha-pinene showed improved behavioral responses and reduced inflammation compared to the hypoxia group (all cases p  < 0.05). This suggests that alpha-pinene may have a protective effect via anti-inflammatory properties against the negative impacts of hypoxia on the developing brain.

Cis-aconitate decarboxylase (CAD), also known as ACOD1 or IRG1, catalyzes the conversion of cis-aconitate to itaconate, playing a pivotal role in innate immunity and inflammatory diseases. Although CAD is a key enzyme in the pathophysiology of inflammatory diseases, no specific inhibitors for CAD currently exist. In this study, we screened 86,326 compounds similar to CAD ligand from the Zinc database to identify potential CAD inhibitors using molecular docking with Molegro Virtual Docker and AutoDock-Vina, followed by molecular dynamics (MD) simulations. The top candidates were further assessed through molecular dynamics (MD) simulations, density functional theory (DFT) calculations, and free energy estimation using the MM/GBSA method. Pharmacokinetics, drug-likeness, and toxicity were evaluated using SWISSADME and Discovery Studio. Among the tested ligands, four compounds demonstrated strong binding affinity, stable interactions, and favorable pharmacokinetic properties, including high gastrointestinal absorption, solubility, and non-toxicity. These compounds demonstrated promising pharmacokinetics, good gastrointestinal absorption, solubility, non-toxicity, and compliance with Lipinski’s rules for drug-like properties. MD simulations further confirmed the stability of ligand-CAD complexes, with cumulative deviations and fluctuations under 2 Å. These findings suggest novel CAD inhibitors with potential as anti-inflammatory agents, paving the way for CAD-targeted drug discovery.

The Non-tuberculous mycobacterial (NTM) isolates should be distinguished from tuberculosis and identified at the species level for choosing an appropriate treatment plan. In this study, two molecular methods were used to differentiate NTM species, including a new designed High Resolution Melting (HRM) and Multilocus Sequence Analysis (MLSA). Seventy-five mycobacterial isolates were evaluated by sequencing four genes ( MLSA) and a HRM assay specifically targeting atpE was designed to rapidly and accurately identify and differentiate mycobacterium species. Out of 70 NTM isolates, 66 (94.3%), 65 (92.9%), 65 (92.9%) and 64 (91.4%) isolates were identified to the species level by PCR of atpE, tuf , rpoB and dnaK genes. We could identify 100% of the isolates to the species level (14 different species) by MLSA. By using HRM assay, all NTM isolates were identified and classified into eight groups, in addition, Mycobacterium tuberculosis and Nocardia were also detected simultaneously. The MLSA technique was able to differentiate all 14 species of NTM isolates. According to the results, the HRM assay is a rapid and beneficial method for identifying NTM, M. tuberculosis (MTB), and Nocardia isolates without sequencing.

Protodioscin (PD) is a natural saponin with anti-inflammatory, anticancer, and metabolic regulatory properties. Senolytic agents, which selectively eliminate senescent cells, are gaining attention for enhancing cancer therapy. This study evaluated the senolytic potential of PD in 3D HepG2 spheroids subjected to Methotrexate (MTX)-induced senescence. Spheroids were generated using the hanging drop method with a 1:1 mixture of HepG2 medium and HFF1-conditioned medium. Senescence was induced with 6 µM MTX, followed by 24-h PD treatment at 1.6, 4, and 8.14 µM (10%, 25%, and 50% of IC₅₀). Cell viability was assessed via MTT; ROS, NO, and TGF-β levels via ELISA; apoptosis via Annexin V/PI; and cell death via Calcein-AM. Expression of p16, p21, p27, p53, and p-Smad2/4 was measured by Western blot, and β-Galactosidase activity was used as a senescence marker. PD dose-dependently reduced spheroid viability and, combined with MTX, significantly enhanced apoptosis, cell death, ROS, NO, β-Gal activity, and expression of p16, p27, and p53, while downregulating p-Smad2/4 and TGF-β compared to MTX treatment alone. These findings suggest that PD potentiates MTX-induced senescence through modulation of cell cycle regulators and inhibition of the TGF-β/p-Smad2/4 pathway, highlighting its potential as a senolytic adjuvant in liver cancer therapy.

In this study, we developed an injectable DNA vaccine targeting the aopB gene of Aeromonas hydrophila , encapsulated within poly(lactic‐co‐glycolic acid) (PLGA) nanoparticles, for use in common carp. Juvenile common carp were divided into six groups with three replicates each. Groups A and B received intramuscular injections of the plasmid containing the target gene (pCDNA3.1‐aopB) with or without encapsulation, respectively. Groups C and D received the plasmid lacking the target gene via the same route. Group E received PLGA, while Group F (control) received phosphate‐buffered saline. Sampling occurred on days 0, 30, and 60, and hematological and immunological indices were compared among the groups. On day 60, all groups were challenged with Aeromonas hydrophila , and cumulative mortality rates were assessed. Our results indicated successful detection of the target gene in various tissues of vaccinated fish. Notably, vaccinated groups exhibited a significant decrease in cumulative mortality ( p < 0.05). Immunological indices, such as serum antibody titer and nonspecific immune responses, significantly improved in vaccinated groups, particularly those receiving the DNA vaccine with PLGA encapsulation ( p   < 0.05). Overall, the DNA vaccine, especially when nanoencapsulated with PLGA, demonstrated efficacy and immunogenicity against A. Hydrophila in common carp, suggesting its potential as a vaccination strategy against this infection. Further research could optimize its effectiveness and applicability.

Potassium diformate (KDF) and calcium diformate (CaDF) are organic acids that modulate growth performance, stress, and biochemical status. So, the present study aimed to investigate the effects of different levels of potassium diformate and calcium diformate on growth performance, stress markers, oxidant/antioxidant status, microbial flora, and some serum biochemical analytes in juvenile Beluga (mean weight: 34 ± 4.5 g). Juvenile Beluga fed control food or diet supplemented with different levels of KDF (1, 1.5, and 2 g/kg) and CaDF (1, 1.5, and 2 g/kg) for 60 days. The results showed that fish fed the 0.15% (1.5 g/kg) KDF showed the favorable growth value at 30 days and relatively less, 0.15% CaDF improved significantly (P<0.05) fish growth performance following 60 days of application (P<0.05). The results showed that dietary administration of KDF and CaDF significantly (P<0.05) increased digestive enzymes. Moreover, elevated biochemical parameters were observed in H. huso fed KDF and CaDF supplemented the diet. Serum oxidant/antioxidant status was significantly (P<0.05) improved in the KDF and CaDF treatments than the control group. Moreover, dietary administration of KDF and CaDF significantly (P<0.05) decreased stress markers of H. huso after 60 days. The liver enzymes activities considerably altered in the KDF and CaDF groups compared with the control group after 60 days. Furthermore, dietary administration of KDF and CaDF significantly (P<0.05) increased intestinal lactic acid bacteria (LAB) of H. huso after 60 days. Based on the results of this study, it appears that incorporating KDF and CaDF into the diet of H. huso can have positive effects on their growth performance and physiological response. The study found that a short-term use of 0.15% (1.5) g/kg of KDF for 30 days was the most effective in promoting growth. However, the potential use of 0.2% (2 g/kg) of KDF and 0.15% (1.5 g/kg) of CaDF for a longer period of 60 days may also be beneficial.