Hindering the biofilm of microbial pathogens and cancer cell lines development using silver nanoparticles synthesized by epidermal mucus proteins from Clarias gariepinus.

Scientists know very little about the mechanisms underlying fish skin mucus, despite the fact that it is a component of the immune system. Fish skin mucus is an important component of defence against invasive infections. Recently, Fish skin and its mucus are gaining interest among immunologists. Characterization was done on the obtained silver nanoparticles Ag combined with Clarias gariepinus catfish epidermal mucus proteins (EMP-Ag-NPs) through UV-vis, FTIR, XRD, TEM, and SEM. Ag-NPs ranged in size from 4 to 20 nm, spherical in form and the angles were 38.10°, 44.20°, 64.40°, and 77.20°, Where wavelength change after formation of EMP-Ag-NPs as indicate of dark brown, the broad band recorded at wavelength at 391 nm. Additionally, the antimicrobial, antibiofilm and anticancer activities of EMP-Ag-NPs was assessed. The present results demonstrate high activity against unicellular fungi C. albicans, followed by E. faecalis. Antibiofilm results showed strong activity against both S. aureus and P. aeruginosa pathogens in a dose-dependent manner, without affecting planktonic cell growth. Also, cytotoxicity effect was investigated against normal cells (Vero), breast cancer cells (Mcf7) and hepatic carcinoma (HepG2) cell lines at concentrations (200-6.25 µg/mL) and current results showed highly anticancer effect of Ag-NPs at concentrations 100, 5 and 25 µg/mL exhibited rounding, shrinkage, deformation and granulation of Mcf7 and HepG2 with IC50 19.34 and 31.16 µg/mL respectively while Vero cells appeared rounded at concentration 50 µg/mL and normal shape at concentration 25, 12.5 and 6.25 µg/ml with IC50 35.85 µg/mL. This study evidence the potential efficacy of biologically generated Ag-NPs as a substitute medicinal agent against harmful microorganisms. Furthermore, it highlights their inhibitory effect on cancer cell lines.

Antibacterial Activity of Green Synthesized Silver Nanoparticles Using Lawsonia inermis Against Common Pathogens from Urinary Tract Infection.

New and creative methodologies for the fabrication of silver nanoparticles (Ag-NPs), which are exploited in a wide range of consumer items, are of significant interest. Hence, this research emphasizes the biological approach of Ag-NPs through Egyptian henna leaves (Lawsonia inermis Linn.) extracts and analysis of the prepared Ag-NPs. Plant extract components were identified by gas chromatography mass spectrometry (GC-mass). The analyses of prepared Ag-NPs were carried out through UV-visible (UV-Vis), X-ray diffraction (XRD), transmission electron microscope (TEM), scanning electron microscope (SEM), and Fourier transform infrared (FTIR) analysis. UV-Vis reveals that Ag-NPs have a maximum peak at 460 nm in visible light. Structural characterization recorded peaks that corresponded to Bragg's diffractions for silver nano-crystal, with average crystallite sizes varying from 28 to 60 nm. Antibacterial activities of Ag-NPs were examined, and it is observed that all microorganisms are very sensitive to biologically synthesized Ag-NPs.

Optimization of oil yield of Pelargonium graveolens L'Hér using Box-Behnken design in relation to its antimicrobial activity and in silico study.

Pelargonium graveolens L'Hér is an important species of genus Pelargonium with an economic value. The unique rose scent of its oil is used in perfume and cosmetic industry. The oil is characterized by the presence of citronellol, geraniol and rose oxide. Fresh aerial parts of P. graveolens at GC-MS analysis of four seasons revealed that autumn constituted the highest yield of the oil. For the first time, optimization of the yield of extracted oil of P. graveolens was performed employing 3-level Box-Behnken design using 3-factors. The GC-MS analysis of the essential oil was performed for the 17-runs. The optimized extraction of the oil was performed employing numerical optimization and studied for antimicrobial, Minimum Inhibitory Concentration (MIC) and biofilm inhibitory activities. The 3 factors followed rank (plant material amount > water volume > NaCl percent in water), in their magnitude of effect on increasing yield of the oil. Increasing the plant material amount increased the yield of the oil by 6-folds compared to NaCl percent in water. The optimized yield of oil (4 ml) was obtained from extraction criteria (150 g of plant, 750 ml of water and 3.585% (26.85 g) of NaCl). Computational docking was performed to overcome the multi-drug resistant Gram-negative bacilli targeting undecaprenyl pyrophosphate synthase (UPPS). The optimized oil exhibited a promising inhibitory activity against Gram-negative bacteria (K. pneumonia and P. aeruginosa) with significant antibiofilm action (P < 0.05). Moreover, it exerted a synergistic effect when combined with various antibiotics (Cefoxitin, Cloxacillin, Oxacillin and Vancomycin) against MRSA clinical strains.

Interception of Epoxide ring to quorum sensing system in Enterococcus faecalis and Staphylococcus aureus.

Quorum sensing inhibitor (QSI) has been attracting attention as anti-virulence agent which disarms pathogens of their virulence rather than killing them. QSI marking cyclic peptide-mediated QS in Gram-positive bacteria is an effective tool to overcome the crisis of antibiotic-dependent chemotherapy due to the emergence of drug resistance strain, e.g., methicillin resistant Staphylococcus aureus (MRSA) and Vancomycin resistant Enterococci (VRE). From a semi-large-scale screening thus far carried out, two Epoxide compounds, Ambuic acid and Synerazol, have been found to efficiently block agr and fsr QS systems, suggesting that the Epoxide group is involved in the mode of action of these QSIs. To address this notion, known natural Epoxide compounds, Cerulenin and Fosfomycin were examined for QSI activity for the agr and fsr systems in addition to in silico and SAR studies. As a result, most of investigated Epoxide containing antibiotics correlatively interfere with QSI activity for the agr and fsr systems under sublethal concentrations.

Recent Insights into Nanotechnology in Colorectal Cancer.

Colorectal cancer (CRC) is the third cancer among the known causes of cancer that impact people. Although CRC drug options are imperfect, primary detection of CRC can play a key role in treating the disease and reducing mortality. Cancer tissues show many molecular markers that can be used as a new way to advance therapeutic methods. Nanotechnology includes a wide range of nanomaterials with high diagnostic and therapeutic power. Several nanomaterials and nanoformulations can be used to treat cancer, especially CRC. In this review, we discuss recent insights into nanotechnology in colorectal cancer.

Assessment of macronutrients dietary intake, central adiposity among pre- and postmenopausal Egyptian women with benign and malignant breast tumors.

BACKGROUND: Breast cancer (BC) is the second most frequent cancer in women and the second most common cancer worldwide. Lifestyle factors, like body weight, physical activity and diet, may be accompanying with higher BC risk. AIM: The assessment of macronutrients dietary intake; protein, fat, carbohydrates and their components of amino, fatty acids, and central obesity/adiposity among pre- and postmenopausal Egyptian women with benign and malignant breast tumors. METHODS: The current case control study included 222 women: 85 control, 54 benign and 83 breast cancer patients. Clinical, anthropocentric and biomedical examinations were performed. Dietary history and health attitude were done. RESULTS: The anthropometric parameters including waist circumference (WC) and the body mass index (BMI) of the benign and the women with malignant breast lesions showed the highest values when compared to the control (35.45 ± 15.58 km2 and 101.24 ± 15.01 cm, 31.39 ± 6.77 km2 and 98.85 ± 13.53 cm and 27.51 ± 7.10 km2 and 84.33 ± 13.78 cm). The biochemical parameters revealed high concentration of the total cholesterol (TC) (192.83 ± 41.54 mg/dl), low density lipoprotein-cholesterol (LDL-C) (117.88 ± 35.18 mg/dl) and the median insulin level 13.8 (10.2-24.1) µu/ml in the malignant patients with high significant difference compared to the control. The malignant patients had the highest daily caloric intake (795.84 ± 519.95 K calories) proteins (65.39 ± 28.77 g), total fats (69.09 ± 32.15 g) and carbohydrates (196.70 ± 85.35 g), when compared to the control. Data also revealed the high daily consumption of the different types of the fatty acids with high linoleic/linoleinic ratio among the malignant group (14.284 ± 6.25). Branched chain amino acids (BGAAs), sulphur amino acids (SAAs), conditional amino acids (CAAs) and aromatic amino acids (AAAs) proved to be the highest in this group. Correlation coefficient between the risk factors revealed either positive or negative weak correlation except that between serum LDL-C concentration and the amino acids (isoleucine, valine cysteine, tryptophan and tyrosine) and negative association with the protective polyunsaturated fatty acids. CONCLUSION: Participants with breast cancer had the greatest levels of body fatness and unhealthy feeding habits relative to their high calorie, protein, carbohydrate, and fat intake.

Advanced approaches for endotoxin detection and removal from snake antivenoms.

A comprehensive assessment of the literature on strategies for the detection and removing endotoxin from biotechnological preparations was conducted. This study highlighted the brief history of endotoxin. After that, a review of endotoxin's chemical and physical features, as well as its pathophysiological consequences when the body is exposed to LPS excessively or systemically, is presented. The procedures for determining endotoxin and the interaction of endotoxin with proteins are also discussed, considering both known approaches and cutting-edge technology in this sector. This review presented the endotoxin detection and removal approaches from antisera with an economical approach using several processes documented in the literature (e.g., adsorption, ultrafiltration, and chromatography). Different methods with relatively high protein recoveries are mentioned. This review concludes that heat activation at 70 °C-80 °C for 10 min and rehydration of the LAL reagent with endotoxin-specific buffer solution is the best technique to control the enhancement problem when testing polyvalent snake venom antiserum samples by the LAL method. The most efficient method for eliminating endotoxins has proven to be affinity resin-based chromatography.

Biosynthesis of Silver and Gold Nanoparticles and Their Efficacy Towards Antibacterial, Antibiofilm, Cytotoxicity, and Antioxidant Activities.

The World Health Organization (WHO) reports that the emergence of multidrug-resistant and the slow advent of novel and more potent antitumor and antimicrobial chemotherapeutics continue to be of the highest concern for human health. Additionally, the stability, low solubility, and negative effects of existing drugs make them ineffective. Studies into alternative tactics to tackle such tenacious diseases was sparked by anticancer and antibacterial. Silver (Ag) and gold (Au) nanoparticles (NPs) were created from Trichoderma saturnisporum, the much more productive fungal strain. Functional fungal extracellular enzymes and proteins carried out the activities of synthesis and capping of the generated nano-metals. Characterization was done on the obtained Ag-NPs and Au-NPs through UV-vis, FTIR, XRD, TEM, and SEM. Additionally, versus methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa, and Klebsiella pneumoniae, the antibacterial activities of Ag-NPs and Au-NPs were assessed. In particular, the Ag-NPs were more effective against pathogenic bacteria than Au-NPs. Furthermore, antibiofilm study that shown Au-NPs had activity more than Ag-NPs. Interestingly, applying the DPPH procedure, these noble metallic NPs had antioxidant activity, in which the IC50 for Ag-NPs and Au-NPs was 73.5 µg/mL and 190.0 µg/mL, respectively. According to the cytotoxicity evaluation results, the alteration in the cells was shown as loss of their typical shape, partial or complete loss of monolayer, granulation, shrinking, or cell rounding with IC50 for normal Vero cell were 693.68 µg/mL and 661.24 µg/mL, for Ag-NPs and Au-NPs, respectively. While IC50 for cancer cell (Mcf7) was 370.56 µg/mL and 394.79 µg/mL for Ag-NPs and Au-NPs, respectively. Ag-NPs and Au-NPs produced via green synthesis have the potential to be employed in the medical industry as beneficial nanocompounds.

Fighting multidrug-resistant Enterococcus faecalis via interfering with virulence factors using green synthesized nanoparticles.

This study aims to synthesize silver nanoparticles by the green method and test it against specific virulence factors in multi-drug resistant Enterococcus faecalis bacteria. virulence factors of E. faecalis clinical isolates were determined and the most potent isolate was selected for further investigations. The prepared Ag-NPs were characterized using UV spectroscopy, FTIR spectroscopy, dynamic light scattering, and transmission electron microscopy (TEM). The result revealed the concentration of 0.0625 mg/ml Ag-NPs was significantly reducing virulence factors in multidrug-resistant E. faecalis without affecting planktonic cell growth. UV-Visible spectroscopy characterization revealed a characteristic surface Plasmon band [SPR] at a wavelength ranging from 256 to 345 and 510 nm in the prepared Ag-NPs. dynamic light scattering indicated it tended to an electrostatic attraction between nanoparticles in the prepared solution. TEM images revealed the average size of Ag-NPs were prepared to be 28.8 nm and the shape was spherical. Green synthesized Ag-NPs have the ability to combat multi-drug-resistant E. faecalis via reducing virulence factors, which is considered a good approach toward resolving the multidrug resistance crisis.

Daphnia magna and Gammarus pulex, novel promising agents for biomedical and agricultural applications.

Various studies have shown the importance of using different types of Zooplankton biomasses as an additional substance in the diet of fish. In addition, the drainage water of the fish cultures could be used in plant irrigation. In this study, biomasses of water flea Daphnia magna and Gammarus pulex collected and tested, for the first time, their effect against pathogenic microorganisms and on plant germination. The results showed significant antibacterial activity of D. magna and G. pulex against Staphylococcus aureus and Pseudomonas aeruginosa bacteria, as well as antifungal activity against Alternaria solani and Penicillium expansum, which gives the possibility to be used as biocontrol against these bacteria and plant pathogenic fungi. Furthermore, both animals showed positive activity in the germination rate of Vicia faba seed, reaching 83.0 ± 3.5 and 86.0 ± 3.8%, respectively. In conclusion, the biomasses of D. magna and G. pulex are promising and effective agents for their use in the medical field against some pathogenic microbes and as stimulators of plant growth.

Molecular docking and nucleotide sequencing of successive expressed recombinant fungal peroxidase gene in E.coli.

BACKGROUND: Fungal peroxidases are oxidoreductases that utilize hydrogen peroxide to catalyze lignin biodegradation. RESULTS: PER-K (peroxidase synthesis codon gene) was transformed from Aspergillus niger strain AN512 deposited in the National Center for Biotechnology Information with the accession number OK323140 to Escherichia coli strain (BL21-T7 with YEp356R recombinant plasmid) via calcium chloride heat-shock method. The impact of four parameters (CaCl2 concentrations, centrifugation time, shaking speed, growth intensity) on the efficacy of the transformation process was evaluated. Furthermore, peroxidase production after optimization was assessed both qualitatively and quantitatively, as well as SDS-PAGE analysis. The optimum conditions for a successful transformation process were as follows: CaCl2 concentrations (50 mM), centrifugation time (20 min), shaking speed (200 rpm), and growth optical density (0.45). PCR and gel electrophoresis detect DNA bands with lengths 175, 179, and 211 bps corresponding to UA3, AmpR, and PER-K genes respectively besides partially sequencing the PER-K gene. Pyrogallol/hydrogen peroxide assay confirmed peroxidase production, and the activity of the enzyme was determined to be 3924 U/L. SDS-PAGE analysis also confirms peroxidase production illustrated by the appearance of a single peroxidase protein band after staining with Coomassie blue R-250. CONCLUSION: A successful peroxidase-gene (PER-K) transformation from fungi to bacteria was performed correctly. The enzyme activity was screened, and partial sequencing of PER-K gene was analyzed successively. The protein 3D structure was generated via in silico homology modeling, and determination of binding sites and biological annotations of the constructed protein were carried out via COACH and COFACTOR based on the I-TASSER structure prediction.

Secondary Metabolites of Actinomycetales as Potent Quorum Sensing Inhibitors Targeting Gram-Positive Pathogens: In Vitro and In Silico Study.

Anti-virulence agents are non-bacteriostatic and non-bactericidal emerging therapeutic options which hamper the production of virulence factors in pathogenic flora. In Staphylococcus aureus and Enterococcus faecalis, regulation of virulence genes' expression occurs through the cyclic peptide-mediated accessory gene regulator (agr) and its ortholog fsr quorum sensing systems, respectively. In the present study, we screened a set of 54 actinomycetales secondary metabolites as novel anti-virulence compounds targeting quorum sensing system of the Gram-positive bacteria. The results indicated that four compounds, Phenalinolactones A-D, BU-4664LMe, 4,5-dehydrogeldamycin, and Questinomycin A, potentially inhibit the agr quorum sensing system and hemolytic activity of S. aureus. On the other hand, Decatromicin A and B, Okilactomycin, Rishirilide A, Abyssomicin I, and Rebeccamycin selectively blocked the fsr quorum sensing system and the gelatinase production in E. faecalis at sub-lethal concentrations. Interestingly, Synerazol uniquely showed the capability to inhibit both fsr and agr quorum sensing systems. Further, in silico molecular docking studies were performed which provided closer insights into the mode of action of these compounds and proposed that the inhibitory activity of these compounds could be attributed to their potential ability to bind to the ATP-active site of S. aureus AgrA. Taken together, our study highlights the potential of actinomycetales secondary metabolites with diverse structures as anti-virulence quorum sensing inhibitors.

Genetic and Histopathological Alterations in Caco-2 and HuH-7 Cells Treated with Secondary Metabolites of Marine fungi.

The present work aimed to study the activity of naturally derived fungal secondary metabolites as anticancer agents concerning their cytotoxicity, apoptotic, genetic, and histopathological profile. It was noticed that Aspergillus terreus, Aspergillus flavus, and Aspergillus fumigatus induced variable toxic potential that was cell type, secondary metabolite type, and concentration dependent. Human colonic adenocarcinoma cells (Caco-2) showed less sensitivity than hepatocyte-derived cellular carcinoma cells (HuH-7), and in turn, the half-maximal inhibitory concentration (IC50) was variable. Also, the apoptotic potential of Aspergillus species-derived fungal secondary metabolites was proven via detection of up-regulated pro-apoptotic genes and down-regulation of anti-apoptotic genes. The expression level was cell type dependent. Concurrently, apoptotic profile was accompanied with cellular DNA accumulation at the G2/M phase, as well as an elevation in Pre-G1 phase but not during G0/G1 and S phases. Also, there were characteristic apoptotic features of treated cells presented as abnormal intra-nuclear eosinophilic structures, dead cells with mixed euchromatin and heterochromatin, ruptured cell membranes, apoptotic cells with irregular cellular and nuclear membranes, as well as peripheral chromatin condensation. It can be concluded that Aspergillus secondary metabolites are promising agents that can be used as supplementary agents to the currently applied anti-cancer drug regimen.

UPLC-ESI-MS/MS profiling of the underground parts of common Iris species in relation to their anti-virulence activities against Staphylococcusaureus.

ETHNOPHARMACOLOGICAL RELEVANCE: The use of plant extracts and their phytochemicals as candidates for targeting the microbial resistance inhibition is increasingly focused in last decades. In Mongolian traditional medicine, Irises were long used for the treatment of bacterial infections. Irises have been used since the Ancient Egyptians. AIM OF THE STUDY: Chemical composition and virulence inhibition potential of both polar (PF) and non-polar fractions (NPF) of three common Iris species (I. confusa, I. pseudacorus and I. germanica) were explored. MATERIAL AND METHODS: Secondary metabolites profiling was characterized by the UPLC-HRMS/MS technique. Multi-variate data analysis was performed using Metaboanalyst 3.0. Anti-virulence inhibitory activity was evaluated via anti-haemolytic assay and Quantitative biofilm inhibition assay. RESULTS: I. pseudacorus PF exhibited the most potent effect against S. aureus haemolytic activity. All the tested fractions from all species, except I. pseudacorus NPF, have no significant inhibition on the biofilm formation of methicillin resistant and sensitive (MRSA and MSSA) S. aureus. I. pseudacorus NPF showed potent biofilm inhibitory potential of 71.4 and 85.8% against biofilm formation of MRSA and MSSA, respectively. Metabolite profiling of the investigated species revealed ninety and forty-five metabolites detected in the PFs and NPFs, respectively. Nigricin-type, tectorigenin-type isoflavonids and xanthones allowed the discrimination of I. pseudacorus PF from the other species, highlighting the importance of those metabolites in exerting its promising activity. On the other hand, triterpene acids, iridals, triacylglycerols and ceramides represented the metabolites detected in highest abundance in I. pseudacorus NPF. CONCLUSIONS: This is the sole map represents the secondary metabolites profiling of the PFs and NPFs of common Iris species correlating them with the potent explored Staphylococcus aureus anti-virulence activity.

Design, synthesis, molecular modeling, and antimicrobial potential of novel 3-[(1H-pyrazol-3-yl)imino]indolin-2-one derivatives as DNA gyrase inhibitors.

A series of 3-[(1H-pyrazol-3-yl)imino]indolin-2-one derivatives were designed using the molecular hybridization method, characterized using different spectroscopic techniques, and evaluated for their in vitro antimicrobial activity. Most of the target compounds demonstrated good to moderate antimicrobial activity compared with ciprofloxacin and fluconazole. Four compounds (8b, 9a, 9c, and 10a) showed encouraging results, with minimal inhibitory concentration (MIC) values (53.45-258.32 µM) comparable to those of norfloxacin (100.31-200.63 µM) and ciprofloxacin (48.33-96.68 µM). Noticeably, the four derivatives revealed excellent bactericidal and fungicidal activities, except for the bacteriostatic potential of compounds 8b and 9a against Escherichia coli and Staphylococcus aureus, respectively. The time-killing kinetic study against S. aureus confirmed the efficacy of these derivatives. Furthermore, two of the four promising derivatives, 9a and 10a, could prevent the formation of biofilms of S. aureus without affecting the bacterial growth at low concentrations. A combination study with seven commercial antibiotics against the multidrug-resistant bacterium P. aeruginosa showed a notable reduction in the antibiotic MIC values, represented mainly through a synergistic or additive effect. The enzymatic assay implied that the most active derivatives had inhibition potency against DNA gyrase comparable to that of ciprofloxacin. Molecular docking and density functional theory calculations were performed to explore the binding mode and study the reactivity of the promising compounds.

Statistical optimization of chromium (VI) reduction using response surface methodology (RSM) by newly isolated Stenotrophomonas sp. (a novel strain).

Isolation of Microorganisms capable of reducing toxic chromium (VI) into less toxic one (Cr (III)) has been given attention due to their significance in bioremediation of the contaminated sites. In the present study, Stenotrophomonas sp. Crt94-4A an isolated strain from tannery wastewater and identified genetically by 16s rRNA gene sequencing was able to grow at concentrations up to 354 mg/L of Cr (VI). The results revealed 1% (w/v) NaCl, 2% (v/v) (2 × 106 CFU) inoculum size, and PH 7 in culture containing glucose and peptone as carbon and nitrogen sources respectively were the best conditions for Cr (VI) reduction. Statistical optimization was performed using Plackett-Burman design where peptone, inoculum size, and NaCl had significant effects on Cr (VI) reduction which were tested by three factors Box-Behnken design (BBD) to determine their correlation. The reduction capacity of Cr (VI) by Stenotrophomonas Sp. Crt94-4A was increased from 82, 55, and 23 to 96, 76, and 45% at 88.5, 177 and 354 mg/L of Cr (VI) respectively, which make this strain a good candidate for bioremediation of Cr (VI).

Development of novel indolin-2-one derivative incorporating thiazole moiety as DHFR and quorum sensing inhibitors: Synthesis, antimicrobial, and antibiofilm activities with molecular modelling study.

Nowadays, it's imperative to develop novel antimicrobial agents active against both drug-sensitive and drug-resistant bacterial infections with favorable profiles as high efficacy, low toxicity, and short therapy duration. Accordingly, a series of new thiazolo-indolin-2-one derivatives were synthesized based on acid and base catalyzed condensation or reaction of thiosemicarbazone 8 with different electrophilic reagents. The structure of the new compounds was confirmed based on elemental analysis and spectral data. Based on the MIC results, the most active thiazolo-indoline derivatives 2, 4, 7a, and 12 exhibited promising antibacterial activity against gram-positive and gram-negative bacteria with weak to moderate antifungal activities. Surprisingly, the N-(thiazol-2-yl)benzenesulfonamide derivative 4 was found to be most active on antibiofilm activity against both S. aureus (ATCC 29213) with BIC50 (1.95 ± 0.01 µg/mL), while 5-(2-oxoindolin-3-ylidene)-thiazol-4(5H)-one derivative 7a exhibited the strongest antibiofilm activity against P. aeruginosa pathogens with BIC50 (3.9 ± 0.16 µg/mL). Further, the thiazole derivatives 2, 4 and 12 exhibited a significant inhibition activity against the fsr system in a dose-dependent manner without affecting bacterial growth. The target derivatives behaved synergistic and additively effect against MDR p. aeruginosa, and thiazole derivative 12 exhibited a high synergistic effect with most tested antibiotics except Cefepime with FIC value ranging between 0.249 and 1.0, reducing their MICs. Interestingly, the 3-(2-(4-thiazol-2-yl)hydrazono)indolin-2-one derivative 12 displayed the highest selectivity to DHFR inhibitory with IC50 value 40.71 ± 1.86 nM superior to those of the reference Methotrexate. Finally, in silico molecular modeling simulation, some physicochemical properties and toxicity predictions were performed for the most active derivatives.

Correlation between phenotypic virulence traits and antibiotic resistance in Pseudomonas aeruginosa clinical isolates.

Pseudomonas aeruginosa is a ubiquitous pathogen capable of infecting virtually all tissues and its one of the standout amongst the most hazardous microorganisms of high morbidity and mortality rates especially in debilitated patients with few successful antibiotic choices available. This pathogen regulating most virulence traits by that so-called quorum sensing (QS), a cell to cell communication system. the present study was intended to phenotypically evaluate the activity of specific virulence traits (including swarming and swimming motility, protease, pyocyanin, and biofilm production) in Pseudomonas aeruginosa clinical isolates and assess the statistical correlation between these traits and antibiotic resistance. One hundred and thirteen bacterial isolates were obtained from different clinical samples and identified as P. aeruginosa, among them, 73.4% have the ability to forming biofilm with different degrees; 59.2% were able to produce pyocyanin pigment while all isolates having the ability to make swarming and swimming motility and able to produce protease enzyme with different degrees. The isolates that produce the higher levels of the virulence traits were identified by both biochemical using Vitek2 automated system and genetically via 16s rRNA gene analysis. The statistical analysis results indicate that a positive significant correlation was found between biofilm formation and other studied virulence traits except for protease (r = 0.584: 0.324, P < 0.05) while a non-significant correlation was found between biofilm formation and protease activity (r = 0.105, P ˃ 0.05). Swimming and swarming motility have a positive significant correlation with other studied virulence traits (r = 0.613: 0.297, P < 0.05) except for protease. Pyocyanin pigment production have a positive significant correlation with other studied virulence traits (r = 0.33: 0.297, P < 0.05) except for protease. on the other hand, negative significant correlations were found between biofilm formation, swimming; and swarming motility, Pyocyanin pigment production, and the susceptibility of antibiotics (r = -0.512: -0.281, P < 0.05). Detection of such correlations in P. aeruginosa is useful for study the behavior of this pathogen and may be provide a new target for the treatment of MDR infections.

Staphylococcus aureus derived hyaluronic acid and bacillus Calmette-Guérin purified proteins as immune enhancers to rabies vaccine and related immuno-histopathological alterations.

PURPOSE: One of the essential goals regarding the successful control of rabies infection is the development of a safe, effective, and inexpensive vaccine. the current study aimed to evaluate the inactivation potential of ß-propiolactone (ßPL), binary ethyleneimine (BEI), and hydrogen peroxide (H2O2). MATERIALS AND METHODS: Estimating the inactivation kinetics of ßPL, BEI, and H2O2 revealed that the tested inactivants could completely and irreversibly inactivate rabies virus within 2, 12, and 4 hours, respectively while maintaining its viral immunogenicity. The potency of ßPL, BEI, and H2O2 inactivated vaccines was higher than the World Health Organization acceptance limit and were in the order of 3.75, 4.21, and 3.64 IU/mL, respectively. Monitoring the humoral and cellular immunity elicited post-immunization using Staphylococcus aureus derived hyaluronic acid (HA) and bacillus Calmette-Guérin purified protein derivative (PPD) adjuvanted rabies vaccine candidates were carried out using enzyme-linked immunosorbent assay. RESULTS: Results demonstrated that both adjuvants could progressively enhance the release of anti-rabies total immunoglobulin G as well as the pro-inflammatory mediators (interferon-gamma and interleukin-5) relative to time. However, a higher immune response was developed in the case of HA adjuvanted rabies vaccine compared to PPD adjuvanted one. The harmful consequences of the tested adjuvants were considered via investigating the histopathological changes in the tissues of the immunized rats using hematoxylin and eosin stain. Lower adverse effects were observed post-vaccination with HA and PPD adjuvanted vaccines compared to that detected following administration of the currently used alum as standard adjuvant. CONCLUSION: Our findings suggested that HA and PPD could serve as a promising platform for the development of newly adjuvanted rabies vaccines with elevated immune enhancing potentials and lower risk of health hazards.

HPLC-PDA-ESI-MS/MS Profiling and Anti-Biofilm Potential of Eucalyptussideroxylon Flowers.

The development of multidrug-resistant bacterial strains is a worldwide emerging problem that needs a global solution. Exploring new natural antibiofilm agents is one of the most important alternative therapies in combating bacterial infections. This study aimed at testing the antimicrobial potential of Eucalyptus sideroxylon flowers extract (ESFE) against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Candida albicans prior to testing the antibiofilm activity against S. aureus, P. aeruginosa and C. albicans. ESFE demonstrated antimicrobial activity and promising inhibition activity against methicillin-resistant S. aureus (MRSA) biofilm formation up to 95.9% (p < 0.05) at a concentration of 0.05 mg/mL and eradicated C. albicans biofilm formation up to 71.2% (p < 0.05) at a concentration of 0.7 mg/mL. LC-MS analysis allowed the tentative identification of eighty-three secondary metabolites: 21 phloroglucinol, 18 terpenes, 16 flavonoids, 7 oleuropeic acid derivatives, 7 ellagic acid derivatives, 6 gallic acid derivatives, 3 phenolic acids, 3 fatty acids and 2 miscellaneous. In conclusion, E. sideroxylon is a rich source of effective constituents that promote its valorization as a promising candidate in the management of multidrug-resistant bacterial infections.