Molecular analysis to identify novel potential biomarkers as drug targets in colorectal cancer therapy: an integrated bioinformatics analysis.

Colorectal cancer (CRC) is a heterogeneous disease that requires new diagnostic and prognostic markers. Integrated bioinformatics approach to identify novel therapeutic targets associated with CRC. Using GEO2R identified DEGs in CRC, and Funrich software facilitated the visualization of DEGs through Venn diagrams. From a total of 114 enhanced DEGs, potential hub genes were further filtered based on their nodal strength and edges using STRING database. To gain insights into the functional roles of these hub genes, gene ontology and pathway enrichment were conducted thorough g: profiler web server. Subsequently, overall survival plots from GEPIA and oncogenic predictive functions like mRNA expressions for stages and nodal metastasis were employed to identify hub genes in CRC patient samples. Additionally, the cBioPortal and HPA databases also revealed genetic alterations and expression levels in these hub genes in CRC patients, further supporting their involvement in colorectal cancer. Gene expression by RT-PCR shows upregulation of hub genes in HT-29 cells. Finally, our integrated bioinformatic analysis revealed that ABCE1, AURKA, HSPD1, PHKA1, CDK4, and YWHAE as hub genes with potential oncogenic roles in CRC. These genes hold promise as diagnostic and prognostic markers for colorectal tumorigenesis, providing insights into targeted therapies for improved patient outcomes.

Biopreservative and Anti-Mycotoxigenic Potentials of Lactobacillus paracasei MG847589 and Its Bacteriocin in Soft White Cheese.

Probiotics and their bacteriocins have increasingly attracted interest for their use as safe food preservatives. This study aimed to produce soft white cheese fortified with Lacticaseibacillus MG847589 (Lb. paracasei MG847589) and/or its bacteriocin; cheese with Lacticaseibacillus (CP), cheese with bacteriocin (CB), and cheese with both Lacticaseibacillus and bacteriocin (CPB) were compared to control cheese (CS) to evaluate their biopreservative and anti-mycotoxigenic potentials for prolonged shelf life and safe food applications. The effects of these fortifications on physiochemical, microbial, texture, microstructure, and sensory properties were studied. Fortification with Lacticaseibacillus (CP) increased acidity (0.61%) and microbial counts, which may make the microstructure porous, while CPB showed intact microstructure. The CPB showed the highest hardness value (3988.03 g), while the lowest was observed with CB (2525.73 g). Consequently, the sensory assessment reflected the panelists' preference for CPB, which gained higher scores than the control (CS). Fortification with Lb. paracasei MG847589 and bacteriocin (CPB) showed inhibition effects against S. aureus from 6.52 log10 CFU/g at time zero to 2.10 log10 CFU/g at the end of storage, A. parasiticus (from 5.06 to 3.03 log10 CFU/g), and P. chrysogenum counts (from 5.11 to 2.86 log10 CFU/g). Additionally, CPB showed an anti-mycotoxigenic effect against aflatoxins AFB1 and AFM1, causing them to be decreased (69.63 ± 0.44% and 71.38 ± 0.75%, respectively). These potentials can extend shelf life and pave the way for more suggested food applications of safe food production by fortification with both Lb. paracasei MG847589 and its bacteriocin as biopreservatives and anti-mycotoxigenic.

Enhanced Nanozymatic Activity on Rough Surfaces for H2O2 and Tetracycline Detection.

The needless use of tetracyclines (TCs) in foodstuffs is a huge health concern in low- and middle-income and Arab countries. Herein, a sensitive and faster monitoring system for H2O2 and TCs is proposed, utilizing the large surface-to-volume ratio of a non-spherical gold nanoparticle/black phosphorus nanocomposite (BP-nsAu NPs) for the first time. BP-nsAu NPs were synthesized through a single-step method that presented nanozymatic activity through 3,3',5,5'-Tetramethylbenzidine (TMB) oxidation while H2O2 was present and obeyed the Michaelis-Menten equation. The nanozymatic activity of the BP-nsAu NPs was enhanced 12-fold and their detection time was decreased 83-fold compared to conventional nanozymatic reactions. The proposed method enabled us to quantify H2O2 with a limit of detection (LOD) value of 60 nM. Moreover, target-specific aptamer-conjugated BP-nsAu NPs helped us detect TCs with an LOD value of 90 nM. The present strategy provides a proficient route for low-level TC monitoring in real samples.

Neoteric Biofilms Applied to Enhance the Safety Characteristics of Ras Cheese during Ripening.

The milk's natural flora, or the starter, can preserve cheesemaking and allow for microbial competition. This investigation aimed to improve cheese safety and assess its characteristics using probiotic cell pellets (LCP) or cell-free extracts (CFS). Cheese samples were collected from different areas to investigate the current contamination situation. Six CFSs of probiotics were assessed as antifungal against toxigenic fungi using liquid and solid media and their aflatoxin reduction impact. The most effective CFS was chosen for cheese coating in nanoemulsion. Coated cheese with CFS, LCP, and LCP-CFS was assessed against control for changes in chemical composition, ripening indications, rheological properties, and microbiology. Results showed significant contamination levels in the collected samples, and toxic fungi were present. Lactobacillus rhamnosus CFS has aflatoxins reducibility in liquid media. During cheese ripening, uncoated cheese showed higher fat, protein, salt content, soluble nitrogen, total volatile fatty acids, tyrosine, and tryptophan contents than coated samples, except for LCP-coating treatment. Cheese rheology indicated that coating treatments had the lowest hardness, cohesiveness, gumminess, chewiness, and springiness compared to uncoated cheese. Uncoated cheese had the highest yeast and mold counts compared to the treated ones. The LCP-CFS-coated cheese showed no Aspergillus cells for up to 40 days. Uncoated Ras cheese recorded slightly lower flavor, body, texture, and appearance scores than coated cheeses. In conclusion, coating cheese with L. rhamnosus nanoemulsion has antifungal and antiaflatoxigenic properties, even for LCP, CFS, and CFS-LCP, which could extend cheese shelf life.

Non-spherical gold nanoparticles enhanced fluorescence of carbon dots for norovirus-like particles detection.

BACKGROUND: Norovirus is a common pathogen that causes foodborne outbreaks every year and the increasing number of deaths caused by it has become a substantial concern in both developed and underdeveloped countries. To date, no vaccines or drugs are able to control the outbreak, highlighting the importance of finding specific, and sensitive detection tools for the viral pathogen. Current diagnostic tests are limited to public health laboratories and/or clinical laboratories and are time-consuming. Hence, a rapid and on-site monitoring strategy for this disease is urgently needed to control, prevent and raise awareness among the general public. RESULTS: The present study focuses on a nanohybridization technique to build a higher sensitivity and faster detection response to norovirus-like particles (NLPs). Firstly, the wet chemical-based green synthesis of fluorescent carbon quantum dots and gold nanoparticles (Au NPs) has been reported. Then, a series of characterization studies were conducted on the synthesized carbon dots and Au NPs, for example, high-resolution transmission emission microscopy, fluorescence spectroscopy, fluorescence life-lime measurement, UV-visible spectroscopy, and X-ray diffraction (XRD). The fluorescence emission of the as-synthesized carbon dots and the absorption of Au NPs were located at 440 nm and 590 nm, respectively. Then, the plasmonic properties of Au NPs were utilized to enhance the fluorescence emission of carbon dots in the presence of NLPs in human serum. Here, the enhanced fluorescence response was linearly correlated up to 1 µg mL-1. A limit of detection (LOD) value was calculated to be 80.3 pg mL-1 demonstrating that the sensitivity of the proposed study is 10 times greater than that of the commercial diagnostic kits. CONCLUSIONS: The proposed exciton-plasmon interaction-based NLPs-sensing strategy was highly sensitive, specific, and suitable for controlling upcoming outbreaks. Most importantly, the overall finding in the article will take the technology a step further to applicable point-of-care (POC) devices.

Prevalence of Aflatoxins in Camel Milk from the Arabian Peninsula and North Africa: A Reduction Approach Using Probiotic Strains.

Camel milk is known as a source of nutritional and health supplements. It is known to be rich in peptides and functional proteins. One main issue facing it is related to its contamination, mainly with aflatoxins. The present study aimed to evaluate camel milk samples from different regions while trying to reduce its toxicity using safe approaches based on probiotic bacteria. Collected samples of camel milk were sourced from two main regions: the Arabic peninsula and North Africa. Samples were tested for their contents of aflatoxins (B1 and M1) using two techniques to ensure desired contamination levels. Additionally, feed materials used in camel foods were evaluated. Applied techniques were also tested for their validation. The antioxidant activity of camel milk samples was determined through total phenolic content and antioxidant activity assays. Two strains of probiotic bacteria (Lactobacillus acidophilus NRC06 and Lactobacillus plantarum NRC21) were investigated for their activity against toxigenic fungi. The result revealed high contamination of aflatoxin M1 for all samples investigated. Furthermore, cross-contamination with aflatoxin B1 was recorded. Investigated bacteria were recorded according to their significant inhibition zones against fungal growth (11 to 40 mm). The antagonistic impacts were between 40% and 70% against toxigenic fungi. Anti-aflatoxigenic properties of bacterial strains in liquid media were recorded according to mycelia inhibition levels between 41 to 52.83% against Aspergillus parasiticus ITEM11 with an ability to reduce aflatoxin production between 84.39% ± 2.59 and 90.4% ± 1.32 from media. Bacteria removed aflatoxins from the spiked camel milk in cases involving individual toxin contamination.

In-Vitro and In-Silico Investigation for the Spent-Coffee Bioactive Phenolics as a Promising Aflatoxins Production Inhibitor.

Aflatoxin, is a naturally occurring polyketide generated by Aspergillus flavus via biosynthetic pathways, including polyketide synthase (PKS) and non-ribosomal enzymes. The in vitro analysis supported by molecular dynamics (MD) techniques was used to examine the antifungal and anti-aflatoxigenic activity of spent coffee grounds (SCGs) methanol extract. The High-Performance Liquid Chromatography results revealed the presence of 15 phenolic acids and five flavonoids. (R)-(+)-Rosmarinic acid (176.43 ± 2.41 µg/g) was the predominant of the detected acids, followed by gallic acid (34.83 ± 1.05 µg/g). At the same time, apigenin-7-glucoside is the dominant flavonoid in the SCGs extract by 1717.05 ± 5.76 µg/g, and naringin (97.27 ± 1.97 µg/g) comes next. The antifungal and anti-aflatoxigenic activity of the SCGs extracts was 380 µL/mL and 460 µL/mL, respectively. The SGGs' effect of inhibiting five Aspergillus strains' growth on the agar media ranged between 12.81 ± 1.71 to 15.64 ± 1.08 mm by two diffusion assays. Molecular docking results confirmed the inhibitory action of different phenolics and flavonoids on the PKS and NPS key enzymes of the aflatoxin biosynthetic mechanism. The SCGs extract components with the highest free binding energy, naringin (-9.1 kcal/mL) and apigenin 7-glucoside (-9.1 kcal/mol), were subjected to an MD simulation study. The computational results infer the stabilizing effects on the enzymes upon ligand binding led to the impairment in its functionality. The current study represents a novel attempt to assess the anti aflatoxins mechanism of phenolics and flavonoids targeting PKS and NPS via computational approaches compared to in-vitro assays.

Quality Attributes of Sesame Butter (Tahini) Fortified with Lyophilized Powder of Edible Mushroom (Agaricus blazei).

Sesame butter (tahini) is a common appetizer and food additive in the Mediterranean basin. Pathogenic strains and mycotoxin content are the most hazardous issues in the final product. This investigation aimed to enhance the quality and safety properties of tahini products against microbial hazards and mycotoxins. Local samples of tahini were evaluated for natural contamination, including mycotoxin level determinations. Agaricus blazei was utilized as a bioactive source and evaluated for the bioactive content of laccase, B-glucan, antioxidant activity, and phenolic content, as well as antimicrobial and antioxidant potency. Two fortification ratios (0.5% and 1.0%) were chosen to apply Agaricus in tahini sesame as a model. Chemical composition, color attributes, sensory properties, emulsion, and oxidative stability were evaluated for the fortified samples versus the control. The results reflected increments of protein (22.91 ± 0.64% to 29.34 ± 0.96%), fiber content (3.09 ± 0.05% to 6.27 ± 0.06%), emulsion stability (84.9 ± 1.24% to 95.41 ± 0.56%), oxidative stability, and bioactive group content. The fortification process is reflected by the absence of Salmonella, Listeria, and E. coli bacteria from contaminated samples after 30 days of storage. The water activity for 1.0% fortification (0.154 ± 0.001) was recorded as lower than the control sample (0.192 ± 0.002). Moreover, the degradation of aflatoxins and zearalenone content was recorded during storage. The degradation ratio reached 68% and 97.2% for 0.5% and 1.0% fortifications, respectively, while zearalenone degradation recorded a decline of 26.7% and 33.7%, respectively, for the same fortification ratios. These results recommended 1.0% lyophilized mushroom fortification as a quality and ameliorative safety treatment for tahini products.

Effect of Cactus (Opuntia ficus-indica) and Acacia (Acacia seyal) Gums on the Pasting, Thermal, Textural, and Rheological Properties of Corn, Sweet Potato, and Turkish Bean Starches.

This study was planned to explore the locally available natural sources of gum hydrocolloids as a natural modifier of different starch properties. Corn (CS), sweet potato (SPS), and Turkish bean (TBS) starches were mixed with locally extracted native or acetylated cactus (CG) and acacia (AG) gums at 2 and 5% replacement levels. The binary mixtures (starch-gums) were prepared in water, freeze dried, ground to powder, and stored airtight. A rapid viscoanalyzer (RVA), differential scanning calorimeter (DSC), texture analyzer, and dynamic rheometer were used to explore their pasting, thermal, textural, and rheological properties. The presence of acetylated AG or CG increased the final viscosity (FV) in all three starches when compared to starch pastes containing native gums. Plain SPS dispersion had a higher pasting temperature (PT) than CS and TBS. The addition of AG or CG increased the PT of CS, SPS, and TBS. The thermograms revealed the overall enthalpy change of the starch and gum blends: TBS > SPS > CS. The peak temperature (Tp) of starches increased with increasing gum concentration from 2 to 5% for both AG and CG native and modified gums. When compared to the control gels, the addition of 2% CG, either native or modified, reduced the syneresis of starch gels. However, further addition (5% CG) increased the gels' syneresis. Furthermore, the syneresis for the first cycle on the fourth day was higher than the second cycle on the eighth day for all starches. The addition of native and acetylated CG reduced the hardness of starch gels at all concentrations tested. All of the starch dispersions had higher G' than G″ values, indicating that they were more elastic and less viscous with or without the gums. The apparent viscosity of all starch gels decreased as shear was increased, with profiles indicating time-dependent thixotropic behavior. All of the starch gels, with or without gums, showed a non-Newtonian shear thinning trend in the shear stress vs. shear rate graphs. The addition of acetylated CG gum to CS resulted in a higher activation energy (Ea) than the native counterparts and the control. More specifically, starch gels with a higher gum concentration (5%) provided greater Ea than their native counterparts.

Characterization of Orange Peel Extract and Its Potential Protective Effect against Aluminum Chloride-Induced Alzheimer's Disease.

Alzheimer's disease (AD) is a devastating neurodegenerative disorder without a cure. Hence, developing an effective treatment or protective agent is crucial for public health. The present study aims to characterize orange peel extract (OPE) through in vitro and in silico studies. Furthermore, it examines the protective effect of OPE against experimentally-induced Alzheimer's disease in rats. The total phenolic and flavonoid content of OPE was 255.86 ± 1.77 and 52.06 ± 1.74 (mg/100 g), respectively. Gallic acid, the common polyphenol in OPE detected by HPLC was 3388.60 µg/100 g. OPE antioxidant IC50 was 67.90 ± 1.05, 60.48 ± 0.91, and 63.70 ± 0.30 by DPPH, ABTS and Hydroxyl radical scavenging activity methods, respectively. In vitro anti-acetylcholinesterase (AChE) IC50 was 0.87 ± 0.025 mg/mL for OPE and 2.45 ± 0.001 mg/mL for gallic acid. Molecular docking analysis for human AChE (4EY7) with donepezil, gallic acid, and acetylcholine showed binding energy ΔG values of -9.47, -3.72, and -5.69 Kcal/mol, respectively. Aluminum chloride injection (70 mg/Kg/day for 6 weeks) induced Alzheimer's-like disease in male rats. OPE (100 and 200 mg/kg/d) and gallic acid (50 mg/kg/d) were administered orally to experimental animals for 6 weeks in addition to aluminum chloride injection (as protective). OPE was found to protect against aluminum chloride-induced neuronal damage by decreasing both gene expression and activity of acetylcholinesterase (AChE) and a decrease in amyloid beta (Aß42) protein level, thiobarbituric acid-reactive substances (TBARS), and nitric oxide (NO), and increased reduced glutathione (GSH) level and activity of the antioxidant enzymes in the brain tissues. Additionally, gene expressions for amyloid precursor protein (APP) and beta secretase enzyme (BACE1) were downregulated, whereas those for presinilin-2 (PSEN2) and beta cell lymphoma-2 (BCL2) were upregulated. Furthermore, the reverse of mitochondrial alternation and restored brain ultrastructure might underlie neuronal dysfunction in AD. In conclusion, our exploration of the neuroprotective effect of OPE in vivo reveals that OPE may be helpful in ameliorating brain oxidative stress, hence protecting from Alzheimer's disease progression.

Boiling Technique-Based Food Processing Effects on the Bioactive and Antimicrobial Properties of Basil and Rosemary.

Rosemary (Rosmarinus officinalis) and basil (Ocimum sanctum Linn) are mostly used as herbal teas, made by steeping whole or ground herbs in boiling water. Hence, it is important to know the effect of boiling time on the bioactivity of these herbs. The effect of different boiling times (5, 10, and 15 min) on the antioxidant and antimicrobial properties, and some selected phenolic compounds of these herbs was examined in this study. Experimental results revealed that basil displayed the highest total polyphenol content (TPC), total flavonoid content (TFC), and antioxidant activity when it was boiled for 5 min, and the lowest TPC was obtained when it was boiled for 15 min. On the other hand, rosemary had the highest TPC, TFC, and antioxidant potential after being boiled for 15 min, while it had the lowest after being boiled for 5 min. There was no growth inhibition of rosemary extracts against gram-negative bacteria, whereas higher growth inhibition was observed against gram-positive bacteria. The MIC and MBC of rosemary ethanolic extract against Listeria monocytogenes were 5 and 5 mg/mL and against B. subtilis were 10 and 10 mg/mL, respectively. While MIC and MBC of methanolic extract against L. monocytogenes were 5 and 5 mg/mL and against Bacillus subtilis were and 5 and 5 mg/mL, respectively. Salicylic acid was the most abundant (324.7 mg/100 g dry weight (dw)) phenolic compound in the rosemary sample boiled for 5 min, and acetyl salicylic acid was the most abundant (122.61 mg/10 g dw) phenolic compound in the basil sample boiled for 15 min.

Development of a novel homogeneous immunoassay using the engineered luminescent enzyme NanoLuc for the quantification of the mycotoxin fumonisin B1.

Compared to the traditional heterogeneous assays, a homogeneous immunoassay is a preferred format for its simplicity. By cloning and isolating luminescent proteins from bioluminescent organisms, bioluminescence has been widely used for various biological applications. In this study, we present the development of a homogeneous luminescence immunoassay (FNanoBiT assay) for detection of fumonisin B1 (FB1), based on the binding of two subunits of an engineered luminescent protein (NanoLuc). For the detection of the mycotoxin FB1 in foods, the anti-fumonisin antibody was conjugated to the large subunit of NanoLuc (FLgBiT), and the FB1 was conjugated to the small subunit (FSmBiT). The conjugates were used for the detection of FB1 in a competitive immunoassay format without the need of a secondary antibody, or washing steps. The developed FNanoBiT assay revealed high specificity toward FB1 with no cross-reactivity with other mycotoxins, and it demonstrated acceptable recovery (higher than 94%) and relative standard deviation from spiked maize samples. Further, the assay was successfully applied for the detection of FB1 in naturally contaminated maize, with a dynamic range of 0.533-6.81 ng mL-1 and a detection limit of 0.079 ng mL-1. The results derived with FNanoBiT assay of all spiked samples showed a strong correlation to those obtained by the High-performance liquid chromatography method. Thus, the FNanoBiT based homogeneous immunoassay could be used as a rapid, and simple tool for the analysis of mycotoxin-contaminated foods.

Rapid detection and differentiation of Staphylococcus colonies using an optical scattering technology.

Staphylococcus species are a major pathogen responsible for nosocomial infections and foodborne illnesses. We applied a laser-based BARDOT (bacterial rapid detection using optical scattering technology) for rapid colony screening and detection of Staphylococcus on an agar plate and differentiate these from non-Staphylococcus spp. Among the six growth media tested, phenol red mannitol agar (PRMA) was found most suitable for building the Staphylococcus species scatter image libraries. Scatter image library for Staphylococcus species gave a high positive predictive value (PPV 87.5-100%) when tested against known laboratory strains of Staphylococcus spp., while the PPV against non-Staphylococcus spp. was 0-38%. A total of nine naturally contaminated bovine raw milk and ready-to-eat chicken salad samples were tested, and BARDOT detected Staphylococcus including Staphylococcus aureus with 80-100% PPV. Forty-five BARDOT-identified bacterial isolates from naturally contaminated foods were further confirmed by tuf and nuc gene-specific PCR and 16S rRNA gene sequence. This label-free, non-invasive on-plate colony screening technology can be adopted by the food industries, biotechnology companies, and public health laboratories for Staphylococcus species detection including S. aureus from various samples for food safety and public health management. Graphical abstract.