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JPMA. the Journal of the Pakistan... Apr 2023To characterise the biofilm matrix composition of a newly described Staphylococcus aureus biofilm phenotype.
OBJECTIVES
To characterise the biofilm matrix composition of a newly described Staphylococcus aureus biofilm phenotype.
METHOD
This experimental study was conducted at the Faculty of Pharmacy, Helwan University, Cairo, Egypt, from January 2021 to March 2022, and comprised methicillin-resistant Staphylococcus aureus and methicillin-susceptible Staphylococcus aureus biofilm-forming clinical isolates which were allowed to construct biofilms under two distinct culture conditions; one a commonly used condition, and the other one a novel, more biologically-relevant condition. The formed biofilms were analysed for matrix composition through treatment with proteinase,sodium meta-periodate, and streptokinase. The efficacy of Cis-2-Decenoic acid and hamamelitannin on the biologically-relevant biofilms was evaluated using biofilm viability assay based on a colorimetric assay for measuring cell metabolic activity and scanning electron microscope imaging. Data was analysed using GraphPad Prism 5.01.
RESULTS
Of the 58 isolates, 45(77.6%) were methicillin-resistant Staphylococcus aureus and 13(22.4%) were methicillin susceptible Staphylococcus aureus. There was significant difference in responses to streptokinase, proteinase and sodium meta-periodate (p<0.05) among the differentially-developed biofilms in methicillin-resistant Staphylococcus aureus isolates. Regarding the methicillin-susceptible Staphylococcus aureus isolates, the differentially-developed biofilms showed significantly different liabilities to streptokinase only (p<0.05). Mean biofilm inhibition for Cis-2- Decenoic acid was 54.27±27.93% and mean biofilm dispersion was 71.92±11.59% while the corresponding valuesfor hamamelitannin were 83.03±13.95% and 70.48±7.116% against the newly described methicillin-resistant Staphylococcus aureus biofilm phenotype.
CONCLUSIONS
Applying biologically-relevant culture conditions on staphylococci biofilms and antibiofilm drugs is recommended.
Topics: Humans; Staphylococcus aureus; Methicillin-Resistant Staphylococcus aureus; Methicillin; Anti-Bacterial Agents; Staphylococcal Infections; Biofilms; Peptide Hydrolases; Phenotype; Streptokinase; Sodium; Microbial Sensitivity Tests
PubMed: 37482852
DOI: 10.47391/JPMA.EGY-S4-34 -
Nutrients May 2023Royal jelly (RJ) is a naturally occurring substance synthesized by honeybees and has various health benefits. Herein, we focused on the medium-chain fatty acids (MCFAs)...
Royal jelly (RJ) is a naturally occurring substance synthesized by honeybees and has various health benefits. Herein, we focused on the medium-chain fatty acids (MCFAs) unique to RJ and evaluated their therapeutic efficacy in treating non-alcoholic fatty liver disease (NAFLD). We examined / mice that were exclusively fed a normal diet, / mice exclusively fed a normal diet, and / mice fed varying RJ quantities (0.2, 1, and 5%). RJ improved NAFLD activity scores and decreased gene expression related to fatty acid metabolism, fibrosis, and inflammation in the liver. RJ regulated innate immunity-related inflammatory responses in the small intestine and decreased the expression of genes associated with inflammation and nutrient absorption transporters. RJ increased the number of operational taxonomic units, the abundance of , and seven taxa, including bacteria that produce short-chain fatty acids. RJ increased the concentrations of RJ-related MCFAs (10-hidroxy-2-decenoic acid, 10-hydroxydecanoic acid, 2-decenedioic acid, and sebacic acid) in the serum and liver. These RJ-related MCFAs decreased saturated fatty acid deposition in HepG2 cells and decreased the gene expression associated with fibrosis and fatty acid metabolism. RJ and RJ-related MCFAs improved dysbiosis and regulated the expression of inflammation-, fibrosis-, and nutrient absorption transporter-related genes, thereby preventing NAFLD.
Topics: Mice; Animals; Bees; Non-alcoholic Fatty Liver Disease; Dysbiosis; Fatty Acids; Inflammation; Fibrosis; Mice, Inbred Strains
PubMed: 37299544
DOI: 10.3390/nu15112580 -
Molecules (Basel, Switzerland) Mar 2023is the causative agent of American foulbrood (AFB), the most serious bacterial disease affecting developing honeybee larvae and pupas. In this study, a library of 24...
is the causative agent of American foulbrood (AFB), the most serious bacterial disease affecting developing honeybee larvae and pupas. In this study, a library of 24 (thio)glycosides, glycosyl sulfones, 6--esters, and ethers derived from d-mannose, d-glucose, and d-galactose having C10 or C12 alkyl chain were evaluated for their antibacterial efficacy against two strains. The efficacy of the tested compounds determined as minimal inhibitory concentrations (MICs) varied greatly. Generally, dodecyl derivatives were found to be more potent than their decylated analogs. Thioglycosides were more efficient than glycosides and sulfones. The activity of the 6--ether derivatives was higher than that of their ester counterparts. Seven derivatives with dodecyl chain linked (thio)glycosidically or etherically at C-6 showed high efficacy against both strains (MICs ranged from 12.5 μM to 50 μM). Their efficacies were similar or much higher than those of selected reference compounds known to be active against -lauric acid, monolaurin, and honeybee larval food components, 10-hydroxy-2-decenoic acid, and sebacic acid (MICs ranged from 25 μM to 6400 μM). The high efficacies of these seven derivatives suggest that they could increase the anti- activity of larval food and improve the resistance of larvae to AFB disease through their application to honeybee colonies.
Topics: Bees; Animals; United States; Paenibacillus larvae; Esters; Sulfides; Anti-Bacterial Agents; Larva; Carbohydrates; Sulfones; Ethers; Glycosides; Paenibacillus
PubMed: 36985490
DOI: 10.3390/molecules28062516 -
Molecules (Basel, Switzerland) Feb 2023Hepatocellular carcinoma (HCC) is the most common form of liver cancer that occurs in hepatocytes. Although many chemical drugs, e.g., cisplatin, methotrexate, taxis,...
BACKGROUND
Hepatocellular carcinoma (HCC) is the most common form of liver cancer that occurs in hepatocytes. Although many chemical drugs, e.g., cisplatin, methotrexate, taxis, and doxorubicin are used to treat HCC, there have been numerous reports related to the side effects of these drugs (e.g., emerging drug resistance, bone marrow failure, and gastrointestinal disorders). These issues led scientists to search for the novel anti-cancer drugs, mainly in natural products with greater efficiency and less toxicity. The current survey was intended to assess the anti-cancer effects of queen bee acid (10-Hydroxy-2-Decenoic Acid, 10-HDA) and its cellular mechanisms against the human hepatoma cell line HepG2.
MATERIALS AND METHODS
The MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay was used to evaluate the effect of 10-HDA on the viability of HepG2 cells. The initial and late apoptosis in the HepG2 cells treated with 10-HDA were assessed by the Annexin-V (AV) assay. The level of the gene and protein expression of some apoptosis genes (e.g., caspase-3, Bcl-2-associated X protein (BAX), and B-cell lymphoma protein 2 (Bcl-2)), Poly (ADP-ribose) polymerases (PARP), and miRNA-34a (miR-34a), were measured by real-time PCR and Western blot.
RESULTS
The obtained findings revealed that HepG2 cell viability was markedly reduced ( < 0.01) following exposure to 10-HDA in a dose-dependent matter. The calculated half maximal cytotoxic concentration (CC50) value of 10-HDA was 59.6 µg/mL for HepG2 cells, while this value for normal THLE-3 cells was 106.4 µg/mL. We found that 10-HDA markedly elevated ( < 0.01) the percentage of necrotic and apoptotic cells from 0.94 to 9.7 and 27.6%, respectively. The real-time PCR results showed that the expression levels of the caspase-3, Bax, and miR-34a genes were significantly ( < 0.001) elevated. Contrary to these results, a significant ( < 0.01) reduction in the expression level of the Bcl2 gene was observed. The levels of protein expression of Caspase-3, PARP, and Bax were markedly elevated following exposure of HepG2 cells to 10-HDA at ¼ CC50, ½ CC50, and CC50. The level of protein expression of Bcl-2 was markedly reduced following exposure of HepG2 cells to 10-HDA at ¼ CC50, ½ CC50, and CC50 ( < 0.01).
CONCLUSION
The current results confirmed the potent in vitro cytotoxic effects of 10-HDA on HepG2 cells with no significant cytotoxic effects on normal cells. Although its mechanisms of action have not been fully studied, the induction of apoptosis via different pathways was determined as one of the principle mechanisms of action of 10-HDA against HepG2 cells. Nevertheless, additional surveys must be performed to clearly understand the mechanisms of action and safety of this fatty acid.
Topics: Humans; Carcinoma, Hepatocellular; Liver Neoplasms; bcl-2-Associated X Protein; Caspase 3; Poly(ADP-ribose) Polymerase Inhibitors; Apoptosis; Proto-Oncogene Proteins c-bcl-2; Antineoplastic Agents; Hep G2 Cells; MicroRNAs
PubMed: 36838959
DOI: 10.3390/molecules28041972 -
Biological & Pharmaceutical Bulletin 2023Royal jelly (RJ), an essential food for the queen honeybee, has a variety of biological activities. Although RJ exerts preventive effects on various lifestyle-related...
Royal jelly (RJ), an essential food for the queen honeybee, has a variety of biological activities. Although RJ exerts preventive effects on various lifestyle-related diseases, such as osteoporosis and obesity, no study evaluated the effect of RJ on the development of osteoarthritis (OA), the most common degenerative joint disease. Here, we showed that daily oral administration of raw RJ significantly prevented OA development in vivo following surgically-induced knee joint instability in mice. Furthermore, in vitro experiments using chondrocytes, revealed that raw RJ significantly reduced the expression of inflammatory cytokines and enzymes critical for the degradation of the extracellular matrix (ECM). Similar results were observed after treatment with 10-hydroxy-2-decenoic acid, the most abundant and unique fatty acid in raw RJ. Our results suggest that oral supplementation with RJ would benefit the maintenance of joint health and prophylaxis against OA, possibly by suppressing the activity of inflammatory cytokines and ECM-degrading enzymes.
Topics: Animals; Bees; Mice; Fatty Acids; Cytokines; Osteoarthritis; Dietary Supplements
PubMed: 36724964
DOI: 10.1248/bpb.b22-00654 -
Current Research in Food Science 2022Royal jelly (RJ) is a popular functional food with a wealth of health-promoting effects. Over 90% of the global RJ is produced in China mainly by a high RJ-producing...
Royal jelly (RJ) is a popular functional food with a wealth of health-promoting effects. Over 90% of the global RJ is produced in China mainly by a high RJ-producing honeybee (RJB) strain that can accept and feed a great number of queen larvae for RJ production. To elucidate RJ changes due to queen cell numbers (QCNs), we compared the yield, larval acceptance rate, metabolic and proteomic profiles, and antioxidant activities of RJ from 1 to 5 strips of queen cells (64 per strip) in RJB colonies. As QCNs increased, the larval acceptance rate was not found to vary ( = 0.269) whereas the RJ weight per cell began to significantly decline in the 5-strip colonies ( < 0.05). Increased QCNs had a profound impact on RJ metabolic profiles and mainly reduced fatty acid levels. Remarkably, the 10-hydroxy-2-decenoic acid (10-HDA) content, a most important indicator of RJ quality, declined gradually from 2.01% in the 1-strip colonies to 1.52% in the 5-strip colonies ( < 0.001). RJ proteomic profiles were minimally altered and antioxidant activities were not significantly changed by QCNs. Collectively, the metabolomics and proteomics data and the antioxidant activity test represent a global evaluation of the quality of RJ produced with different QCNs. Our findings gain new insights into higher-quality RJ production using the high-yielding RJBs.
PubMed: 36254242
DOI: 10.1016/j.crfs.2022.10.014 -
IScience Oct 2022The G protein-coupled receptor 84 (GPR84) is found in immune cells and its expression is increased under inflammatory conditions. Activation of GPR84 by medium-chain...
The G protein-coupled receptor 84 (GPR84) is found in immune cells and its expression is increased under inflammatory conditions. Activation of GPR84 by medium-chain fatty acids results in pro-inflammatory responses. Here, we screened available vertebrate genome data and found that GPR84 is present in vertebrates for more than 500 million years but absent in birds and a pseudogene in bats. Cloning and functional characterization of several mammalian GPR84 orthologs in combination with evolutionary and model-based structural analyses revealed evidence for positive selection of bear GPR84 orthologs. Naturally occurring human GPR84 variants are most frequent in Asian populations causing a loss of function. Further, we identified - and -2-decenoic acid, both known to mediate bacterial communication, as evolutionary highly conserved ligands. Our integrated set of approaches contributes to a comprehensive understanding of GPR84 in terms of evolutionary and structural aspects, highlighting GPR84 as a conserved immune cell receptor for bacteria-derived molecules.
PubMed: 36164652
DOI: 10.1016/j.isci.2022.105087 -
Cancer Biology & Medicine Aug 2022This study aimed at examining the alterations in metabolomic profiles caused by treatment of infection, and the associations between key plasma metabolites and the risk...
OBJECTIVE:
This study aimed at examining the alterations in metabolomic profiles caused by treatment of infection, and the associations between key plasma metabolites and the risk of gastric lesion progression during follow-up after treatment.
METHODS:
An intervention trial was performed in 183 participants, 117 of whom were positive participants receiving treatment for infection. positive participants were prospectively followed for 182 to 1,289 days. Untargeted metabolomics assays were conducted on plasma samples collected at baseline, 6 months after treatment, and during continued follow-up.
RESULTS:
We identified 59 metabolites with differential posttreatment changes between participants with successful and failed eradication, 17 metabolites significantly distinguished participants with successful failed eradication. Two metabolites [PC(18:1(11Z)/14:1(9Z)) and (2S)-6-amino-2-formamidohexanamide] showed posttreatment changes positively associated with successful eradication, and were inversely associated with the risk of gastric lesion progression among participants with successful eradication. In contrast, 9-decenoic acid showed posttreatment changes inversely associated with successful eradication: its level was positively associated with the risk of gastric lesion progression among participants with successful eradication. Although the identified metabolites showed a temporary but significant decline after treatment, the trend generally reversed during continued follow-up, and pretreatment levels were restored.
CONCLUSIONS:
Treatment of infection significantly altered plasma metabolic profiles in the short term, and key metabolites were capable of distinguishing participants with successful failed eradication, but might not substantially affect metabolic regulation in the long term. Several plasma metabolites were differentially associated with the risk of gastric lesion progression among participants with successful or failed eradication.
Topics: Follow-Up Studies; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Prospective Studies
PubMed: 36069529
DOI: 10.20892/j.issn.2095-3941.2022.0255 -
Biomaterials Advances May 2022Graphene oxide quantum dots (GOQDs) have attracted substantial attention in numerous fields due to their unique physicochemical properties. However, their nanotoxicity...
Graphene oxide quantum dots (GOQDs) have attracted substantial attention in numerous fields due to their unique physicochemical properties. However, their nanotoxicity and potential for use in biomedicine still require further study. In this work, the effects of GOQD and trans- 10-hydroxy-2-decenoic acid (10-HDA) cotreatment on the immune function of macrophages (RAW264.7 cells) were investigated. In particular, LC/MS-based metabolomics was performed to evaluate the effects of GOQDs on the metabolism of LPS-stimulated macrophages. Herein, we fabricated GOQDs with good dispersibility and a uniform size distribution of approximately 7 nm using a polyimide-pyrolyzed carbon film as the working electrode, a high-voltage graphite electrode as the cathode, and HO as the oxidant. The GOQDs entered the macrophages and emitted green fluorescence under UV irradiation. Cotreatment with GOQDs and 10-HDA induced RAW 264.7 cell proliferation. GOQDs promoted the anti-inflammatory effect of 10-HDA on LPS-stimulated RAW264.7 cells and attenuated the secretion of TNF-α, IL-6, and IL-1β. The metabolites in RAW264.7 cells treated with GOQDs were significantly different from those in RAW264.7 cells treated with LPS. The enrichment analysis showed that treatment with GOQDs interfered with amino acid metabolism, and lipid metabolism. Our results demonstrate the role of GOQDs in macrophages and provide a basis for their further application in biomedical fields.
Topics: Anti-Inflammatory Agents; Fatty Acids, Monounsaturated; Graphite; Hydrogen Peroxide; Lipopolysaccharides; Macrophages; Quantum Dots
PubMed: 35929313
DOI: 10.1016/j.bioadv.2022.212774 -
Food Technology and Biotechnology Jun 2022Acquisition of migratory potential is pivotal for cancer cells, enabling invasion and metastasis of colorectal carcinoma. Royal jelly and its bioactive component...
RESEARCH BACKGROUND
Acquisition of migratory potential is pivotal for cancer cells, enabling invasion and metastasis of colorectal carcinoma. Royal jelly and its bioactive component -10-hydroxy-2-decenoic acid (10H2DA) showed remarkable antimetastatic potential, but the molecular mechanism underlying this activity is unclear.
EXPERIMENTAL APPROACH
Identification and quantification of 10H2DA in royal jelly originating from Serbia was done by HPLC method. Cytotoxicity of 10H2DA was measured by tetrazolium dye MTT test in concentration range 1-500 μg/mL after 24 and 72 h. Its effect on the collective and single-cell migration was measured by wound healing and transwell migration assays. Invasive potential of cancer cells was evaluated by a transwell method modified with collagen. Immunofluorescence was used for migratory and invasive protein expression, while the gene expression of these markers was evaluated by quantitative real time polymerase chain reaction (qRT-PCR). All assays were applied on human colorectal carcinoma HCT-116 and SW-480 cell lines and, except for MTT, evaluated after 24 h of treatment with two selected concentrations of royal jelly and 10H2DA.
RESULTS AND CONCLUSIONS
According to HPLC, the mass fraction of 10H2DA in royal jelly was 0.92% (/). Treatment with 10H2DA showed no cytotoxic effect; however, significant inhibitory potential of royal jelly and 10H2DA on the motility and invasiveness of colorectal cancer cells was observed. More pronounced effect was exerted by 10H2DA, which significantly suppressed collective cell migration and invasiveness of SW-480 cells, as well as single- and collective cell migration and invasive potential of HCT-116 cell line. Treatments increased epithelial markers E-cadherin and cytoplasmic β-catenin in HCT-116 cells, thus stabilizing intercellular connections. In SW-480 cells, 10H2DA increased E-cadherin on protein and gene level, and suppressed epithelial-mesenchymal transition (EMT) markers. In both cell lines, treatments induced significant suppression of promigratory/proinvasive markers: N-cadherin, vimentin and Snail on protein and gene level, which explains decreased migratory and invasive potential of HCT-116 and SW-480 cells.
NOVELTY AND SCIENTIFIC CONTRIBUTION
Our study presents new findings and elucidation of royal jelly and 10H2DA molecular mechanism that underlies their antimigratory/antiinvasive activity on colorectal cancer cells. These findings are shown for the first time indicating that these natural products are a valuable source of anticancer potential and should be reconsidered for further antitumour therapy.
PubMed: 35910272
DOI: 10.17113/ftb.60.02.22.7495