AMB Express最新文献

Process and culture medium composition in bioreactor could be optimized in order to find the best conditions that improve survival of probiotic microorganism under exposure to gastric conditions such low pH and bile salts. Therefore, this study aimed to optimize agitation, yeast extract, and di-sodium phosphate (Na₂HPO₄) concentration to improve the survival under gastric conditions of a multistrain consortium produced in a laboratory bioreactor. Viability, survival low pH (3.00), bile salt tolerance, and antagonistic activity against the pathogen Streptococcus agalactiae were evaluated. As the main result, a high concentration of di-sodium phosphate (2.63% w/v) increased the viability of L. lactis A12 (9.05 to 9.46 Log₁₀ CFU/mL) and Priestia species (0.00 to 6.88 Log₁₀ CFU/mL), survival to pH 3.00 (60 to 93%), survival of bile salts (58- 93%) antagonistic activity (8.74 to 15.56 mm), and final pH of culture medium (4.34 to 6.95). Optimal conditions that improved probiotics characteristics were 150 RPM, 0.83% w/v yeast extract, and 2.63% w/v Na₂HPO₄. Co-culture of L. lactis A12 with Priestia species improved significantly (p < 0.05) the antagonistic activity (10.41 mm) against S. agalactiae compared to mono-culture (7.70 mm). Our results suggested that was possible to produce a potential multistrain preparation in a lab bioreactor with high viability of L. lactis A12 (9.33 Log₁₀ CFU/mL), high survival to gastric conditions (> 85%), and with antagonistic activity against fish pathogen. This preparation could be used as a feed additive intended for fish nutrition.

Cryptococcal pneumonia is a severe fungal infection of the respiratory system, predominantly caused by Cryptococcus neoformans. Its incidence is increasing, driven by evolving pathogen dynamics and heightened susceptibility among patient populations. This investigation aimed to assess the combined therapeutic efficacy of Fluconazole and Amphotericin B for cryptococcal pneumonia and to explore the roles of miR-15b and TGF-β1 in modulating treatment response. Twenty-eight patients diagnosed with cryptococcal pneumonia were randomly allocated to receive either Amphotericin B monotherapy (control group) or a combination of Amphotericin B and Fluconazole (observation group) over a 14-day period. Key respiratory function indices-forced expiratory volume in one second (FEV1), forced vital capacity (FVC), and FEV1/FVC ratios-were measured pre- and post-treatment, alongside levels of procalcitonin (PCT), soluble triggering receptor expressed on myeloid cells-1 (sTREM-1), miR-15b, and TGF-β1. Results demonstrated marked improvements in pulmonary function within the observation group, with significantly higher FEV1, FVC, and FEV1/FVC values than those in the control group (P < 0.05). Additionally, the observation group exhibited greater reductions in PCT and sTREM-1, coupled with notable elevations in miR-15b and TGF-β1 levels. The combination therapy achieved a total response rate of 92.86%, significantly surpassing the control's 57.14% efficacy (P < 0.05). These findings indicate that Fluconazole combined with Amphotericin B not only enhances clinical efficacy by mitigating inflammation but also supports lung function recovery with a favourable safety profile, highlighting its utility in managing cryptococcal pneumonia effectively.

Candida albicans is a commensal fungus that naturally inhabits the vagina. However, overgrowth of C. albicans can result in vulvovaginal candidiasis (VVC), one of the most prevalent fungal infections affecting women. The rapid emergence of azole resistance in C. albicans, in addition to the limited available antifungal agents, complicates the treatment and emphasizes the urgent need for novel therapeutic options. Efflux-mediated azole resistance is a common resistance mechanism in fluconazole (FLZ)-resistant C. albicans. Combination therapy using natural compounds is a potential approach that can restore fluconazole's antifungal activity in azole-resistant isolates via efflux pump inhibition. This study aimed to evaluate the ability of celastrol, a natural triterpene, to retrieve FLZ antifungal activity against azole-resistant C. albicans in vitro and in vivo. Celastrol did not exhibit antifungal activity against the tested clinical isolates; however, the sub-MIC of celastrol inhibited rhodamine 6G (R6G) efflux and increased R6G accumulation inside celastrol-treated C. albicans cells. Synergy was spotted between celastrol and FLZ via a checkerboard assay. Quantification of m-RNA levels of efflux-mediated azole resistance genes within azole-resistant C. albicans demonstrated CDR1 overexpression. Upon celastrol treatment, a significant decline in ABC transporters transcript levels were detected. Moreover, molecular docking demonstrated that celastrol is a potential ABC efflux transporters blocker that successfully fits into target binding pockets. A negligible hemolytic effect of celastrol against human erythrocytes was observed. In the in vivo model of VVC, the combination of FLZ and celastrol in vaginal gel revealed a drastic reduction in the fungal burden with apparently normal vaginal tissue. Celastrol promising in vitro and in vivo findings strengthen its future use for the treatment of azole-resistant C. albicans.

The urgent need to address the growing problem of antimicrobial resistance in multidrug-resistant bacteria requires the development of pioneering approaches to treatment. The present study aims to evaluate the antimicrobial potential of the essential oils (EOs) of Moringa oleifera (moringa), Cinnamomum verum (cinnamon), and Nigella sativa (black seed) and the synergistic effect of the mixture of these oils against Staphylococcus aureus MCC 1351. Statistical modeling revealed cinnamon oil had the highest individual antimicrobial potency, followed by black seed oil. The combination of the three EOs exhibited significant synergistic effects compared to the individual oils, with a Fractional Inhibitory Concentration (∑FIC) index of 0.27. L-Optimal mixture design of response surface methodology (RSM) identified the optimal mixture as moringa: cinnamon: black seed oils by the ratio of (1:1:1) in run 15 (0.338:0.307:0.355 mL) (v/v). This mixture exhibited significant antibacterial efficacy, outperforming individual oils and conventional antibiotics like tetracycline. Specifically, the combination reduced the MIC values from 3.12, 0.78, and 6.25 to 0.25, 0.06, and 0.78 μg/mL for moringa, cinnamon, and black seed oil, respectively. Synergistic interactions between oils further boosted efficacy, with moringa-cinnamon and cinnamon-black seed pairings exhibiting the strongest synergies. The developed predictive models for IZD and MIC showed excellent fit, with R² values of 0.9843 and 0.9958, respectively. Pareto chart analysis highlighted the predominant individual and synergistic effects, with the Moringa-Cinnamon interaction exhibiting the highest positive synergy. Notably, the oil mixture of run 15 demonstrated excellent biocompatibility, maintaining 97.6% viability of normal human skin fibroblasts (HSF) after 24 h exposure to 200 μL EOs of the mixture per mL. Gas chromatography mass spectrometry (GC/MS) identified abundant bioactive phytochemicals like cinnamaldehyde, linoleic acid, and palmitic acid methyl esters underlying the observed antimicrobial effects. This rationally designed, synergistic phytochemical combination presents a promising natural therapeutic against antibiotic-resistant S. aureus while exhibiting minimal cytotoxicity. The results underscore how combining essential oils could help address the issue of antibiotic resistance in S. aureus.

The increasing demand for natural alternatives to synthetic fungicides has prompted research into natural products like essential oils for postharvest disease management. This study investigated the antifungal, antioxidant, cytotoxic, and genotoxic potential of essential oil mixtures derived from oregano, rosemary, and mint against Penicillium digitatum, the predominant fungal pathogen causing green mold in orange fruits. P. digitatum NPAGRASU 2024 was isolated and identified as the most abundant species (60.2%) from infected oranges. It was deposited in GenBank with gene accession number PP930644 and deposited in MIRCEN culture collection as EMCC 358874. The antimicrobial activity of individual essential oils was evaluated, with oregano exhibiting the highest antifungal activity (inhibition zone diameter of 4.2 cm) against P. digitatum. L-optimal mixture design of response surface methodology (RSM) optimization revealed a highly effective mixture (Run 8) comprising 46.26% oregano and 53.74% rosemary, with a 99.65% actual growth reduction. The oregano oil demonstrated potent antioxidant activity, reaching approximately 75% DPPH radical scavenging at 3.125 mg/mL. Cytotoxicity assessment using the MTT assay showed morphological changes and reduced cell viability in liver cells treated with the PEOs mixture at 300 µg/mL. However, the optimized mixture did not induce significant chromosomal aberrations compared to the control, suggesting minimal genotoxic effects. In vivo, evaluation on oranges revealed 60% inhibition of green mold by the 1% (v/v) optimized mixture for 7 days. Histological analysis indicated low toxicity to the liver at the highest tested concentration (1% mixture). GC-MS analysis identified major compounds like cavarcol, caryophyllene, eucalyptol, phenols and levomenthol in oregano oil, contributing to its bioactivities. This study demonstrates the potential of optimized essential oil mixtures as effective and eco-friendly alternatives for postharvest disease control.

Phyllostachys makinoi, an endemic bamboo species in Taiwan, is underutilized, despite its rich forest resources. Known for its antioxidant, anti-inflammatory, and antibacterial properties, this study explores the antimicrobial, anti-inflammatory, and wound-healing activities of P. makinoi extracts. The antibacterial potential of P. makinoi extracts was first evaluated using the agar diffusion method, along with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays. Subsequently, electron microscopy and a conductivity meter were employed to assess whether P. makinoi extracts exert antibacterial effects by disrupting bacterial cell structures. Finally, the anti-inflammatory and cell proliferation-promoting effects of P. makinoi extracts were assessed in RAW264.7 and CCD-1112Sk cell models. The MIC and MBC of the P. makinoi water extracts against two multidrug-resistant Staphylococcus aureus strains were 4 and 16-32 mg/mL, respectively, while those for ethanol extracts were 2 and 32 mg/mL, respectively. In the time-kill assay, both strains were killed after treatment with extracts for 12 and 18 h. Scanning electron microscopy revealed that the bacterial cells treated with the extracts appeared rough, ruptured, and shriveled. The extracts disrupted the cell membranes, causing electrolyte, protein, and nucleic acid leakage, leading to cell death. Additionally, P. makinoi extracts reduced lipopolysaccharide-induced nitric oxide, tumor necrosis factor-α, interleukin (IL)-1β, and IL-6 levels in RAW264.7 cells, and promoted wound healing by enhancing CCD-1112Sk cell proliferation. In conclusion, P. makinoi water and ethanol extracts demonstrated antibacterial and anti-inflammatory effects, showing potential for treating wound infections and accelerating healing, making them promising candidates for wound-healing therapies.

Cryptosporidium sp. is an obligatory intracellular apicomplexan protozoan parasite that causes a disease called cryptosporidiosis with substantial veterinary and medical importance. Therefore, this study aimed to evaluate an early diagnosis of cryptosporidiosis using the anti-Cryptosporidium parvum oocyst immunoglobulin IgG polyclonal antibodies (anti-C. parvum IgG PAbs)-based sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of Cryptosporidium oocyst antigens in fecal samples of farm animals in Egypt. Further molecular identification and sequencing were performed for the detected isolates. Eight hundred and twenty fecal samples of farm animals; 102 buffalo calves, 120 cattle calves, 100 lambs and 98 goat kids, 80 buffaloes, 60 cattle, 160 sheep and 100 goats, collected from different small-scale farms and local holders were examined for cryptosporidiosis by Modified Ziehl-Neelsen (MZN) technique. The percentage of positivity was 45.1%, 50%, 20%, 18.4%, 31.25%, 38.3%, 18.8%, and 11% in buffalo calves, cattle calves, lambs, goat kids, adult buffaloes, adult cattle, sheep, and goats, respectively. Molecular identification of Cryptosporidium samples was performed based on COWP gene, revealing the isolates: GenBank: OQ121955.1, OR029973.1 and PP316107.1 which were identical to the C. parvum and GenBank: PP316108.1 and OR029972.1 which were identical to C. hominis and C. andersoni, respectively. Then, C. parvum oocysts were used for preparation of antigens and rabbit immunization. Anti-C. parvum IgG PAbs were purified and characterized by SDS-PAGE and then labeled with horseradish peroxidase (HRP). Anti-C. parvum IgG PAbs in-house sandwich ELISA was prepared, then tested this ELISA on 820 samples and compared results with MZN microscopical examination and a commercial sandwich ELISA kit. In this study, in-house sandwich ELISA scored higher sensitivity of 98%, 100% specificity, validity 99% and relative agreement 98.6% than (92%, 90%, 91% and 91.4%) of MZN and (96%, 95%, 95.5% and 95.7%) of coproantigen commercial sandwich ELISA kit, respectively. Moreover, we used PCR to evaluate the positivity of in-house sandwich ELISA results, and the total PCR positive samples were 263 out of 268 sandwich ELISA positive samples (98.13%). In conclusion, the prepared Anti-C. parvum IgG PAbs based sandwich ELISA offered a simple and accurate diagnostic method for cryptosporidiosis in the fecal samples of different species of farm animals in Egypt with high sensitivity (98%) and specificity (100%). Further studies on this Anti-C. parvum IgG PAbs may help also in the protection against cryptosporidiosis.

Chemotherapeutic therapies for cancer are frequently associated with cytotoxic side effects that can be harmful to human health, including the development of intestinal mucositis (IM). It mostly affects the gastrointestinal tract, causing ulceration, inflammation, and the formation of lesions in the colon. Surprisingly, despite the frequency of IM, therapeutic choices remain restricted. In our search for new intestinal mucositis therapies, we wanted to see how Lentinan (LT), derived from Lentinus edodes, would fare in mouse models of intestinal mucositis. To create the intestinal mucositis model in mice, we gave them intra-peritoneal doses of 5-fluorouracil (5-FU) (50 mg/kg) and then tested the effects of Lentinan on intestinal mucositis. This examination required constant monitoring of several factors, such as body weight fluctuations, food consumption, and diarrhea. In addition, we measured the levels of certain inflammatory cytokines (Tumour Necrosis Factor-alpha (TNF-α), Interleukin-1 (IL-1), Interleukin-6 (IL-6), and Interleukin-10 (IL-10), looked at the expression of tight junction proteins (Zonula Occludens-1(ZO-1), Claudin-1), measured mucin-2 levels, and looked into changes in the gut flora. In the mouse model of intestinal mucositis, our findings showed that LT effectively reduced weight loss, increased food intake, and relieved diarrhea. Concurrently, we saw a decrease in the expression of inflammatory cytokines such as TNF-α, IL-1, and IL-6, as well as a considerable increase in the concentration of IL-10. Furthermore, LT reduced intestinal mucositis by increasing the length and structural integrity of the colon. Furthermore, increased expression of tight junction proteins (ZO-1, Claudin-1), mucin-2, and an increase in the number of goblet cells all confirmed our previous findings. Notably, the makeup of beneficial bacteria in the stomach increased as well. Finally, our findings suggest that LT can effectively prevent 5-fluorouracil-induced intestinal mucositis in mice by improving immune function, restoring intestinal barrier integrity, and rebalancing gut microbial flora.