Lowest Minimum Inhibitory Concentration Research Articles

Staphylococcus aureus small colony variants: A potentially underestimated microbiological challenge in peritoneal dialysis

IntroductionPeritonitis remains the major infectious complication in the setting of peritoneal dialysis (PD). Despite known only moderate pathogenicity, the most frequently detected pathogens in PD-related peritonitis are surprisingly coagulase-negative staphylococci. However, this could be explained, at least in part, by Staphylococcus aureus small colony variants (SCVs) induced by PD fluids (PDFs) and misidentified by routinely used microbiological methods. Material and methodsBacteria were exposed to commonly used PDFs in various regimens designed to simulate daily use as closely as possible. Wild-type isolates and SCVs were subsequently used to determine minimum inhibitory concentrations (MICs), in vitro biofilm formation capacities, and auxotrophies. Underlying genetic alterations were investigated using whole-genome sequencing, and various microbial identification methods were tested to determine their performance for wild-types and SCVs. ResultsStable SCVs could be isolated most successfully after exposure to glucose-containing PDFs alone. The reading of MICs was significantly affected by the reduced growth of SCVs, resulting in lower MIC values in 44% of all tests. Nonsynonymous mutations were found in all but one SCV, while only two isolates showed typical auxotrophic responses. While MALDI-TOF, PCR and Pastorex Staph-Plus correctly identified all S. aureus SCVs, API-Staph and VITEK-2 yielded identification rates of only 40% and 10%, respectively. ConclusionsOverall, the present study has shown that commercially available PDFs induce S. aureus SCVs in vitro, which are difficult to identify and test for antimicrobial susceptibility and can potentially lead to recurrent or persistent infections. Thus, they represent a potentially underappreciated challenge not only for microbiologists, but also for clinicians.

DEVELOPMENT OF THE PHARMACEUTIC CRYSTALLIZATION PROCESS USING PLASMODIUM FALCIPARUM, A DERIVATIVE OF THIOSEMICARBAZONE

Objective: The current study was designed with the goal of analyzing novel derivatives of organic and semi-organic NLO molecules that are just entering this field as the foundation for future therapeutic development. Methods: Thiosemicarbazones (TSC) are semicarbazide analogs that have sulfur atoms in place of oxygen atoms. The greatest number of therapeutic treatments for various disorders are also being used carbonyl compounds. These two types of organic compounds were combined, and their replacement with crystal growth procedures using solution growth was tested as an antibiotic for a few fastidious and nonfastidious species. The preparation of meta-substituted benzaldehydes using TSC. Tests were conducted with this single component. The following experimental technique was used to determine the antibacterial activity of Plasmodium falciparum. Half-inhibitory maximum (IC50) by calculation technique, cell viability % calculation method, and MTT assay by colorimetric method. Results: Method using agar discs. Using the agar disc diffusion method to measure the inhibition zone width and comparing the minimal inhibitory concentration (MIC) and inhibition zone width of available antibiotics against the aforementioned organism using data from the EUCAST and NCCLS databases, The absolute, relative, and mound slope values of an antibiotic that is currently on the market may be determined using the GraphPad Prism software; the epidemiological cutoff value can be determined using the ECOF Finder software; and the nature of the antibiotic can be determined using the WHONET 5.6 software. The newly developed antibiotics’ size and structure are contrasted with the structures of commercially available antibiotics and living organisms. Conclusion: The design of new drugs using novel derivatives of organic and semiorganic NLO molecules is based on the in vitro method of analysis, which has recently been introduced to this field. P. falciparum has an extremely low MIC value (0.625 μg/mL) against TSCMNB. The MIC break points are always larger than, not equal to, the MIC values and fall within the range of 110 μg/mL–130 μg/mL. We concluded that all of the novel compounds exhibit a susceptible form of inhibition.

In vitro activity of tedizolid against 43 species of Nocardia species

The purpose of the present study was to evaluate the in vitro activity of tedizolid against several clinically significant species of Nocardia by comparing with that of linezolid. A total of 286 isolates of Nocardia species, including 236 clinical isolates recovered from patients in Japan and 50 strains (43 species) purchased from NITE Biological Resource Center, were studied. Antimicrobial susceptibility testing was performed using the broth microdilution method. For the 286 Nocardia isolates, the minimal inhibitory concentration (MIC)50 and MIC90 values of tedizolid were 0.25 and 0.5 μg/ml, and those of linezolid were 2 and 2 μg/ml, respectively. The distribution of the linezolid/tedizolid ratios (MICs of linezolid/MICs of tedizolid) showed that tedizolid had four- to eight-fold higher activity than linezolid in 96.1% (275/286) of Nocardia isolates. Both the tedizolid and linezolid MIC90 values for Nocardia brasiliensis were two-fold higher than those for the other Nocardia species. Both tedizolid and linezolid had low MIC values, 0.25–1 μg/ml and 0.5–4 μg/ml, respectively, even against nine isolates (five species) that were resistant to trimethoprim/sulfamethoxazole. One Nocardia sputorum isolate showed reduced susceptibility to tedizolid (4 μg/ml). Bioinformatics analysis suggests different resistance mechanisms than the oxazolidinone resistance seen in enterococci and staphylococci.

Safety Assessment of Levilactobacillus brevis KU15006: A Comprehensive Analysis of its Phenotypic and Genotypic Properties.

Levilactobacillus brevis KU15006, isolated from kimchi, exhibits pathogen-antagonistic and anti-diabetic activities; however, the safety of this strain has not been assessed. In the present study, L. brevis KU15006 was evaluated to elucidate its safety as a probiotic strain using phenotypic and genotypic analyses. Its safety was assessed using a minimum inhibitory concentration test comprising nine antibiotics, 26 antibiotic resistance genes, a single conjugative element, virulence gene analysis, hemolysis, cell cytotoxicity, mucin degradation, and toxic metabolite production. L. brevis KU15006 exhibited equal or lower minimum inhibitory concentration for the nine antibiotics than the cut-off value established by the European Food Safety Authority. It did not harbor antibiotic resistance and virulence genes. L. brevis KU15006 lacked β-hemolysis, mucin degradation, cytotoxicity against Caco-2 cells, gelatin liquefaction, bile salt deconjugation, and toxic metabolite production abilities. Based on the results, L. brevis KU15006, which has antagonistic and anti-diabetic effects, could be marketed as a probiotic in the future.

Green synthesis and characterization of silver nanoparticles from Ducrosia flabellifolia Boiss. aqueous extract: Anti-quorum sensing screening and antimicrobial activities against ESKAPE pathogens.

Biosynthesis of silver nanoparticles using natural compounds derived from plant kingdom is currently used as safe and low-cost technique for nanoparticles synthesis with important abilities to inhibit multidrug resistant microorganisms (MDR). ESKAPE pathogens, especially MDR ones, are widely spread in hospital and intensive care units. In the present study, AgNPs using Ducrosia flabellifolia aqueous extract were synthesized using a reduction method. The green synthesized D. flabellifolia-AgNPs were characterized by UV-Vis spectrophotometer, Scanning electron microscopy (SEM), and X-ray diffraction assays. The tested D. flabellifolia aqueous extract was tested for its chemical composition using Liquid Chromatography-Electrospray Ionization-Mass Spectrometry (LC-ESI-MS). Anti-quorum sensing and anti-ESKAPE potential of D. flabellifolia-AgNPs was also performed. Results from LC-ESI-MS technique revealed the identification of chlorogenic acid, protocatechuic acid, ferulic acid, caffeic acid, 2,5-dihydroxybenzoic acid, and gallic acid as main phytoconstituents. Indeed, the characterization of newly synthetized D. flabellifolia-AgNPs revealed spherical shape with mean particle size about 16.961±2.914 nm. Bio-reduction of silver was confirmed by the maximum surface plasmon resonance of D. flabellifolia-AgNPs at 430 nm. Newly synthetized D. flabellifolia-AgNPs were found to possess important anti-ESKAPE activity with low minimal inhibitory concentrations (MICs) ranging from 0.078 to 0.312 mg/mL, and low minimal bactericidal concentrations (MBCs) varying from 0.312 to 0.625 mg/mL. Moreover, D. flabellifolia-AgNPs were active against Candida utilis ATCC 9255, C. tropicalis ATCC 1362, and C. albicans ATCC 20402 with high mean diameter of growth inhibition at 5 mg/mL, low MICs, and minimal fungicidal concentrations values (MFCs). The newly synthetized D. flabellifolia-AgNPs were able to inhibit violacein production in Chromobacterium violaceum, pyocyanin in Pseudomonas aeruginosa starter strains. Hence, the newly synthesized silver nanoparticles using D. flabellifolia aqueous extract can be used as an effective alternative to combat ESKAPE microorganisms. These silver nanoparticles can attenuate virulence of Gram-negative bacteria by interfering with the quorum sensing system and inhibiting cell-to-cell communication.

Anti-bacterial Effects of Allium atroviolaceum Hydroalcoholic Extract on Oral Bacteria of Streptococcus viridans Groups

: In light of the escalating global concern over antibiotic resistance, this study aimed to evaluate the antimicrobial activity of the hydroalcoholic extract of Allium atroviolaceum and compare it with 0.2% chlorhexidine using the well diffusion, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) methods against Streptococcus mitis (ATCC 6249), Streptococcus mutans (PTCC 16836), Streptococcus sanguinis (PTCC 1449), and Streptococcus salivarius (PTCC 1448). The well diffusion method revealed the inhibitory effect of the extract, with the highest activity observed against Streptococcus salivarius. Subsequently, MIC and MBC values were determined, indicating that the hydroalcoholic extract had MIC and MBC values of 3.12 mg/mL against Streptococcus sanguinis and 6.25 mg/mL against other strains (Streptococcus mitis, Streptococcus mutans, Streptococcus sanguinis, and Streptococcus salivarius). Chlorhexidine exhibited significantly lower MIC and MBC values of 0.0003%. Based on literature reviews, Allium species exhibit antimicrobial properties. The findings highlight the potential of Allium atroviolaceum extract as an alternative or complementary antimicrobial agent and warrant further investigation to identify its bioactive components and explore their mechanisms of action. This study contributes to understanding natural antimicrobial agents and their potential applications against bacterial infections.

Antifungal Susceptibility Profile of Clinical and Environmental Isolates of Aspergillus Species From a Tertiary Care Center in North India.

Aspergillus species are ubiquitously found in the environment worldwide and are important causative agents for infection. Drug resistance among Aspergillus species is emerging, hencethe present study was undertaken to look for antifungal susceptibility profiles of clinical and environmental isolates of Aspergillus species. During the period from January 2018 to June 2019, a total of 102 Aspergillus isolates (40 clinical, 40 hospital, and 22 community environment) were tested for antifungal susceptibility testing for determination of minimum inhibitory concentration (MIC)/minimum effective concentration (MEC) as per Clinical and Laboratory Standards Institute(CLSI) M38-A3 method for itraconazole, voriconazole, amphotericin B, and caspofungin. Out of these 102 Aspergillus isolates, A. flavus was the most common species present. Aspergillus species were found to have low MIC values to azoles such as itraconazole and voriconazole except for one clinical isolate, which showed a MIC value of 2 μg/ml to voriconazole. Two isolates were non-wild-type for amphotericin B, but all isolates were wild-type for caspofungin. Antifungal susceptibility testing among clinical Aspergillus isolates and environmental surveillance studies in view of emerging drug resistance should be undertaken at a larger scale.

Mechanistic insight into the membrane disrupting properties of thymol in Candida species

Candida infection is a major problem in immunocompromised patients and has become a significant public health concern in recent years due to high toxicity, multidrug resistance and unfavourable side effects of conventional antifungal drugs. This study aims to investigate the antifungal efficacy of thymol, a natural monoterpenoid, on the membrane integrity of Candida albicans, Candida glabrata and Candida tropicalis. Minimum inhibitory concentrations (MICs) of thymol were 32 µg/ml, 63 µg/ml, and 125 µg/ml for C. albicans, C. tropicalis and C. glabrata, respectively. The minimum fungicidal concentrations (MFC) were 64 µg/ml, 110 µg/ml and 240 µg/ml, respectively. At MIC, thymol treated cells showed complete suppression of cell growth (growth kinetics, disc diffusion), altered morphology (scanning electron microscopy), leakage of intracellular macromolecules (absorption at 260 nm), significant reduction in ergosterol levels and inhibition of plasma membrane H+-ATPase (Pma1) activity. The disruption of the fungal membranes by thymol was further confirmed by propidium iodide uptake using flow cytometry and confocal microscopy. In silico studies with membrane-bound proteins (lanosterol 14-α demethylase (LDM) and Pma1), showed good binding affinity of −6.3 kcal/mol and −6.7 kcal/mol, respectively, and formed hydrogen bonds with crucial amino acid residues at the active site of target proteins. With low MIC values, negligible host toxicity, desirable ADME properties, thymol has immense potential as a potent antifungal drug that can be used for developing mechanism-based drugs as it has specific binding affinity for specific target proteins. Studies using animal models are required for further validation.

Antioxidant and antimicrobial activities of mango peel and radish peel-a comparative investigation

The present study investigated the antioxidant and antimicrobial activities of Bao mango and daikon radish peel (Thai variety) ethanolic extracts. The percentage extraction yield of radish peel was higher compared to the mango peel (p < 0.05). However, Mango peel extract (MPE) contained higher total phenolic content (TPC) than radish peel extract (RPE) (p < 0.05). Similar to the TPC value, antioxidant activities including DPPH radical scavenging activity, reducing power, and peroxidation inhibition were higher in the MPE, compared to the RPE, except for metal chelating activity. The antimicrobial analyses suggested that the MPE showed lower minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values compared to the RPE (p < 0.05) against Gram-positive and Gram-negative bacteria. High-performance liquid chromatography (HPLC) revealed that mangiferin was the major phenolic compound present in MPE, followed by 3, 4 dihydroxybenzoic acid. Therefore, MP could be used as a promising natural antioxidant and antimicrobial agent for food applications.

Antibiotic Combination to Effectively Postpone or Inhibit the In Vitro Induction and Selection of Levofloxacin-Resistant Mutants in Elizabethkingia anophelis.

Fluoroquinolones are potentially active against Elizabethkingia anophelis. Rapidly increased minimum inhibitory concentrations (MICs) and emerging point mutations in the quinolone resistance-determining regions (QRDRs) following exposure to fluoroquinolones have been reported in E. anophelis. We aimed to investigate point mutations in QRDRs through exposure to levofloxacin (1 × MIC) combinations with different concentrations (0.5× and 1 × MIC) of minocycline, rifampin, cefoperazone/sulbactam, or sulfamethoxazole/trimethoprim in comparison with exposure to levofloxacin alone. Of the four E. anophelis isolates that were clinically collected, lower MICs of levofloxacin were disclosed in cycle 2 and 3 of induction and selection in all levofloxacin combination groups other than levofloxacin alone (all p = 0.04). Overall, no mutations were discovered in parC and parE throughout the multicycles inducted by levofloxacin and all its combinations. Regarding the vastly increased MICs, the second point mutations in gyrA and/or gyrB in one isolate (strain no. 1) occurred in cycle 2 following exposure to levofloxacin plus 0.5 × MIC minocycline, but they were delayed appearing in cycle 5 following exposure to levofloxacin plus 1 × MIC minocycline. Similarly, the second point mutation in gyrA and/or gyrB occurred in another isolate (strain no. 3) in cycle 4 following exposure to levofloxacin plus 0.5 × MIC sulfamethoxazole/trimethoprim, but no mutation following exposure to levofloxacin plus 1 × MIC sulfamethoxazole/trimethoprim was disclosed. In conclusion, the rapid selection of E. anophelis mutants with high MICs after levofloxacin exposure could be effectively delayed or postponed by antimicrobial combination with other in vitro active antibiotics.

Mango Peels as an Industrial By-Product: A Sustainable Source of Compounds with Antioxidant, Enzymatic, and Antimicrobial Activity.

Plant waste materials are important sources of bioactive compounds with remarkable health-promoting benefits. In particular, industrial by-products such as mango peels are sustainable sources of bioactive substances, with antioxidant, enzymatic, and antimicrobial activity. Appropriate processing is essential to obtain highly bioactive compounds for further use in generating value-added products for the food industry. The objective of the study was to investigate and compare the biological activity of compounds from fresh and dried mango peels obtained by different conventional methods and unconventional extraction methods using supercritical fluids (SFE). The highest total phenolic content (25.0 mg GAE/g DW) and the total content of eight phenolic compounds (829.92 µg/g DW) determined by LC-MS/MS were detected in dried mango peel extract obtained by the Soxhlet process (SE). SFE gave the highest content of proanthocyanidins (0.4 mg PAC/g DW). The ethanolic ultrasonic process (UAE) provided the highest antioxidant activity of the product (82.4%) using DPPH radical scavenging activity and total protein content (2.95 mg protein/g DW). Overall, the dried mango peels were richer in bioactive compounds (caffeic acid, chlorogenic acid, gallic acid, catechin, and hesperidin/neohesperidin), indicating successful preservation during air drying. Furthermore, outstanding polyphenol oxidase, superoxide dismutase (SOD), and lipase activities were detected in mango peel extracts. This is the first study in which remarkable antibacterial activities against the growth of Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa) and Gram-positive bacteria (Bacillus cereus and Staphylococcus aureus) were evaluated by determining the microbial growth inhibition rate after 12 and 24 h incubation periods for mango peel extracts obtained by different methods. Ethanolic SE and UAE extracts from dried mango peels resulted in the lowest minimum inhibitory concentrations (MIC90) for all bacterial species tested. Mango peels are remarkable waste products that could contribute to the sustainable development of exceptional products with high-added value for various applications, especially as dietary supplements.

Antibacterial activity of plant-derived compounds and cream formulations against canine skin bacteria

An urgent need to find alternative antimicrobial compounds effective in the prevention and treatment of skin infections led us to study the inhibitory activity of eight plant-derived bioactive compounds (betulin, curcumin, glycyrrhizic acid, guaiazulene, piperine, quercetin, quinine, tannic acid) against 14 canine skin isolates (11 Gram-positive and three Gram-negative bacteria) selected based on antibiotic resistance and virulence features. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) were determined using the broth microdilution method. In detail, the results for the eight different plant compounds showed their inhibitory activity in the concentration range from 0.04 to more than 16 mg/ml (MIC) and from 0.25 to more than 16 mg/ml (MBC). The most potent compounds appear to be tannic acid, followed by quinine and curcumin (MIC 0.04–16.0 mg/ml). The most susceptible strain to the tested agents in general was Bacillus cereus AE13, while Enterococcus faecium AA14 was the most resistant strain (the highest MICs) among the tested bacteria. The two most potent plant-derived compounds (tannic acid and quinine) were tested in mixture in different ratios (1:1, 1:2, 2:1). The lowest MIC and MBC values were observed for the 1:2 ratio, which was used for preparation of creams with different cream bases. One of the cream formulations (cream F) was effective up to 63.0 mg/ml (MIC) with a microbial inactivation time of 1–6 h according to the tested strain. This study provides evidence that some plant-derived compounds could have an antimicrobial effect against canine skin bacteria, the strength of which is bacterial strain dependent.

Citrus sinensis Seed-Mediated Gold Nanoparticles for Combating Diabetes and Bacterial Infections: A Promising Multifunctional Nano Formulation

This investigation explores the successful synthesis and comprehensive characterization of Citrus sinensis seed-mediated gold nanoparticles designated as C-AuNPs. Visual confirmation of synthesis was achieved through a distinct ruby red color change in the reaction mixture, followed by UV-Visible spectroscopy, which revealed a characteristic peak absorbance at 522 nm due to the Surface Plasmon Resonance (SPR) phenomenon associated with gold nanoparticles. Transmission Electron Microscopy (TEM) analysis demonstrated uniform, spherical nanoparticles with an average size of 19 nm, ideal for biomedical applications. Dynamic Light Scattering (DLS) revealed a hydrodynamic size of 55 nm in the hydrated state, while a negative zeta potential of −17 mV indicated colloidal stability. C-AuNPs exhibited significant antibacterial activity against both Gram-negative and Gram-positive bacterial strains, surpassing the performance of levofloxacin. Lower Minimum Inhibitory Concentration (MIC) values against bacterial strains further supported their enhanced efficacy, potentially due to interactions with bacterial cell membranes. Furthermore, C-AuNPs demonstrated potent inhibition of α-amylase and α-glucosidase enzymes involved in glucose metabolism, with IC50 values comparable to the anti-diabetic drug acarbose. This suggests their potential as anti-diabetic agents, with the ability to regulate blood sugar levels. C-AuNPs exhibit unique optical, structural, and electrostatic properties, making them promising candidates for diverse biomedical applications. Their small size, stability, antibacterial efficacy, and anti-diabetic properties position C-AuNPs as valuable assets in the realm of nanomedicine and therapeutics, warranting further exploration and development.

Susceptibility of some Enterobacteriaceae isolated from 4 different aquatic environments in DR Congo (Central Africa), to Amoxicillin/Clavulanic acid and some 3rd generation Cephalosporins

Little is known about the variation of the antibiotic susceptibility of different bacterial strains of the same cells species, isolated from different aquatic environments. The present study aims to evaluate the susceptibility towards some 3rd generation cephalosporins (Ceftriaxon, Ceftazidim and Cefotaxim) and Amoxicillin/Clavulanic acid, of Enterobacteriaceae strains isolated from groundwater, stream, hospital wastewater and slaughterhouse wastewater. Enterobacteria species were isolated on Mac Conkey agar, then identified using enzymatic and MALDI-TOF MS system. The antimicrobial susceptibility was carried out using the disk diffusion method. The antibiotic minimum inhibitory concentrations (MICs) were carried using the VITEK®2 system. Bacterial species mostly identified were Klebsiella pneumoniae, Escherichia coli, Salmonella typhi and Ewingelia americana. The antibiotic inhibition diameters and the MICs varied depending on the antibiotics, bacterial species and type of aquatic environment hosting the microorganism. Relatively lower MICs were recorded with Cefotaxim against different bacteria in slaughterhouse wastewater, in river water and in well water. Antibiotic resistance was noted with all strains from hospital wastewater. Significant differences (P˂0.05) amongst antibiotic inhibition diameter was noted for K. pneumoniae and S. typhi in most cases. The relative variations in the action mechanisms amongst antibiotics and the intrinsic defense potential of each bacterial strain, as well as the potential influence of the physicochemical properties of each water medium, could partly be at the origin of the relative differences observed at the phenotypic level. It seems necessary to explore the diversity, similarities and differences amongst antibiotic resistance genes in these different types of aquatic biotopes.

Elucidation of unusual biosynthesis and DnaN-targeting mode of action of potent anti-tuberculosis antibiotics Mycoplanecins

DNA polymerase III sliding clamp (DnaN) was recently validated as a new anti-tuberculosis target employing griselimycins. Three (2 S,4 R)−4-methylproline moieties of methylgriselimycin play significant roles in target binding and metabolic stability. Here, we identify the mycoplanecin biosynthetic gene cluster by genome mining using bait genes from the 4-methylproline pathway. We isolate and structurally elucidate four mycoplanecins comprising scarce homo-amino acids and 4-alkylprolines. Evaluating mycoplanecin E against Mycobacterium tuberculosis surprisingly reveals an excitingly low minimum inhibition concentration at 83 ng/mL, thus outcompeting griselimycin by approximately 24-fold. We show that mycoplanecins bind DnaN with nanomolar affinity and provide a co-crystal structure of mycoplanecin A-bound DnaN. Additionally, we reconstitute the biosyntheses of the unusual l-homoleucine, l-homonorleucine, and (2 S,4 R)−4-ethylproline building blocks by characterizing in vitro the full set of eight enzymes involved. The biosynthetic study, bioactivity evaluation, and drug target validation of mycoplanecins pave the way for their further development to tackle multidrug-resistant mycobacterial infections.

A promising antifungal lipopeptide from Bacillus subtilis: its characterization and insight into the mode of action.

Emerging resistance of fungal pathogens and challenges faced in drug development have prompted renewed investigations into novel antifungal lipopeptides. The antifungal lipopeptide AF3 reported here is a natural lipopeptide isolated and purified from Bacillus subtilis. The AF3 lipopeptide's secondary structure, functional groups, and the presence of amino acid residues typical of lipopeptides were determined by circular dichroism, Fourier transform infrared spectroscopy, and nuclear magnetic resonance spectroscopy. The lipopeptide's low minimum inhibitory concentrations (MICs) of 4-8mg/L against several fungal strains demonstrate its strong antifungal activity. Biocompatibility assays showed that~ 80% of mammalian cells remained viable at a 2 × MIC concentration of AF3. The treated Candida albicans cells examined by scanning electron microscopy, transmission electron microscopy, and atomic force microscopy clearly showed ultrastructural alterationssuch as the loss of the cell shape and cell membrane integrity. The antifungal effect of AF3 resulted in membrane permeabilization facilitating the uptake of the fluorescent dyes-acridine orange (AO)/propidium iodide (PI) and FUN-1. Using 1,6-diphenyl-1,3,5-hexatriene (DPH) and 4-(2-[6-(dioctylamino)-2-naphthalenyl] ethenyl)-1-(3-sulfopropyl) pyridinium inner salt (di-8-ANEPPS), we observed that the binding of AF3 to the membrane bilayer results in membrane disruption and depolarization. Flow cytometry analyses revealed a direct correlation between lipopeptide activity, membrane permeabilization (~ 75% PI uptake), and reduced cell viability. An increase in 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescence demonstrates endogenous reactive oxygen species production. Lipopeptide treatment appears to induce late-stage apoptosis and alterations to nuclear morphology, suggesting that AF3-induced membrane damage may lead to a cellular stress response. Taken together, this study illustrates antifungal lipopeptide's potential as an antifungal drug candidate. KEY POINTS: • The studied lipopeptide variant AF3 displayed potent antifungal activity against C. albicans • Its biological activity was stable to proteolysis • Analytical studies demonstrated that the lipopeptide is essentially membranotropic and able to cause membrane dysfunction, elevated ROS levels, apoptosis, and DNA damage.

Indole clubbed 2,4-thiazolidinedione linked 1,2,3-triazole as a potent antimalarial and antibacterial agent against drug-resistant strain and molecular modeling studies.

In the face of escalating challenges of microbial resistance strains, this study describes the design and synthesis of 5-({1-[(1H-1,2,3-triazol-4-yl)methyl]-1H-indol-3-yl}methylene)thiazolidine-2,4-dione derivatives, which have demonstrated significant antimicrobial properties. Compared with the minimum inhibitory concentrations (MIC) values of ciprofloxacin on the respective strains, compounds 5a, 5d, 5g, 5l, and 5m exhibited potent antibacterial activity with MIC values ranging from 16 to 25 µM. Almost all the synthesized compounds showed lower MIC compared to standards against vancomycin-resistant enterococcus and methicillin-resistant Staphylococcus aureus strains. Additionally, the majority of the synthesized compounds demonstrated remarkable antifungal activity, against Candida albicans and Aspergillus niger, as compared to nystatin, griseofulvin, and fluconazole. Furthermore, the majority of compounds exhibited notable inhibitory effects against the Plasmodium falciparum strain, having IC50 values ranging from 1.31 to 2.79 μM as compared to standard quinine (2.71 μM). Cytotoxicity evaluation of compounds 5a-q on SHSY-5Y cells at up to 100 μg/mL showed no adverse effects. Comparison with control groups highlights their noncytotoxic characteristics. Molecular docking confirmed compound binding to target active sites, with stable protein-ligand complexes displaying drug-like molecules. Molecular dynamics simulations revealed dynamic stability and interactions. Rigorous tests and molecular modeling unveil the effectiveness of the compounds against drug-resistant microbes, providing hope for new antimicrobial compounds with potential safety.

Avoiding the Danger of Rising Resistance in Cutibacterium acnes: Criticality of Benzoyl Peroxide and Antibiotic Fixed Combinations

Background: Antibiotic resistance is a global concern, with several countries reporting resistance in >50% of Cutibacterium acnes (C. acnes) strains. Combination formulations containing an antibiotic and the antimicrobial benzoyl peroxide (BPO) may reduce this resistance risk, especially with prolonged use. This four-part study tested susceptibility of 31 C. acnes clinical strains to antibiotics alone or in combination with BPO.
 Methods: Part 1: C. acnes susceptibility to single-drug antibiotics was assessed via minimum inhibitory concentration (MIC) values obtained from epsilometer tests, with lower MIC indicating higher susceptibility. Part 2: Susceptibility to fixed-dose antibiotic/BPO combination products (branded/in-development) was determined by measuring zone of inhibition using agar diffusion method, with larger diameter indicating increased bacterial inhibition. Part 3: The effect (synergistic, additive, antagonistic, or neutral) of combining clindamycin with BPO on C. acnes inhibition was evaluated using a checkerboard assay, wherein two test compounds are combined in varying concentrations. Part 4: Development of resistance was assessed using serial passage of bacterial cultures in increasing concentrations of clindamycin alone or in combination with BPO.
 Results: Part 1: All antibiotics tested—clindamycin, doxycycline, erythromycin, and minocycline—had similar activity. Susceptibility was highly strain dependent, as some C. acnes strains had elevated MIC—an indication of resistance—against different antibiotics. Part 2: For the 6 C. acnes strains that had no inhibitory zone (0 cm) with clindamycin alone, formulations with BPO enhanced activity against the same isolates (range: 0.8-2.2 cm with clindamycin phosphate 1.2%/adapalene 0.15%/BPO 3.1%, clindamycin phosphate 1.2%/BPO 5%, or clindamycin phosphate 1.2%/BPO 3.75%). Part 3: The combination of clindamycin and BPO resulted in an additive effect for 4 of the 7 acne-associated strains tested and was neutral for 3 strains. Part 4: Bacterial cultures repeatedly exposed to a combination of clindamycin and BPO did not develop resistance to C. acnes, which occurred with exposure to antibiotic alone.
 Conclusions: Overall, antibiotic susceptibility was highly strain dependent and antibiotic formulations with BPO exhibited enhanced activity against less susceptible C. acnes strains. Fixed combinations of BPO with an antibiotic may improve antimicrobial activity and protect against resistance development.
 
 Funding: Ortho Dermatologics.

Chemical Composition, Antioxidant, Anticancer, and Antibacterial Activities of Roots and Seeds of Ammi visnaga L. Methanol Extract.

For centuries, plants and their components have been harnessed for therapeutic purposes, with Ammi visnaga L. (Khella) being no exception to this rich tradition. While existing studies have shed light on the cytotoxic and antimicrobial properties of seed extracts, there remains a noticeable gap in research about the antimicrobial, antioxidant, and anticancer potential of root extracts. This study seeks to address this gap by systematically examining methanol extracts derived from the roots of A. visnaga L. and comparing their effects with those of seed extracts specifically against breast cancer cells. Notably, absent from previous investigations, this study focuses on the comparative analysis of the antimicrobial, antioxidant, and anticancer activities of both root and seed extracts. The methanol extract obtained from A. visnaga L. seeds demonstrated a notably higher level of total phenolic content (TPC) than its root counterpart, measuring 366.57 ± 2.86 and 270.78 ± 2.86 mg GAE/g dry weight of the dry extract, respectively. In the evaluation of antioxidant activities using the DPPH method, the IC50 values for root and seed extracts were determined to be 193.46 ± 17.13 μg/mL and 227.19 ± 1.48 μg/mL, respectively. Turning our attention to cytotoxicity against breast cancer cells (MCF-7 and MDA-MB-231), both root and seed extracts displayed similar cytotoxic activities, with IC50 values of 92.45 ± 2.14 μg/mL and 75.43 ± 2.32 μg/mL, respectively. Furthermore, both root and seed extracts exhibited a noteworthy modulation of gene expression, upregulating the expression of caspase and Bax mRNA levels while concurrently suppressing the expression of anti-apoptotic genes (Bcl-xL and Bcl-2), thereby reinforcing their potential as anticancer agents. A. visnaga L. seed extract outperforms the root extract in antimicrobial activities, exhibiting lower minimum inhibitory concentrations (MICs) of 3.81 ± 0.24 to 125 ± 7.63 μg/mL. This highlights the seeds' potential as potent antibacterial agents, expanding their role in disease prevention. Overall, this study underscores the diverse therapeutic potentials of A. visnaga L. roots and seeds, contributing to the understanding of plant-derived extracts in mitigating disease risks.

In vitro characterization and molecular epidemiology of Cryptococcus spp. isolates from non-HIV patients in Guangdong, China.

The burden of cryptococcosis in mainland China is enormous. However, the in vitro characterization and molecular epidemiology in Guangdong, a key region with a high incidence of fungal infection in China, are not clear. From January 1, 2010, to March 31, 2019, clinical strains of Cryptococcus were collected from six medical centres in Guangdong. The clinical information and characteristics of the strains were analysed. Furthermore, molecular types were determined. A total of 84 strains were collected, mostly from male and young or middle-aged adult patients. Pulmonary and cerebral infections (82.1%) were most common. All strains were Cryptococcus neoformans, grew well at 37°C and had capsules around their cells. One melanin- and urea- and one melanin+ and urea- variants were found. Although most strains exhibited a low minimum inhibitory concentration (MIC) value for voriconazole (mean: 0.04 μg/mL) and posaconazole (mean: 0.12 μg/mL), the results for these isolates showed a high degree of variation in the MIC values of fluconazole and 5-fluorocytosine, and resistance was observed for 4 out of 6 drugs. A significant proportion of these strains had MIC values near the ECV values, particularly in the case of amphotericin B. The proportion of strains near the clinical breakpoints was as follows: fluconazole: 3.66%; voriconazole: 3.66%; itraconazole: 6.10%; posaconazole: 13.41%; amphotericin B: 84.15%; 5-fluorocytosine: 2.44%. These strains were highly homogeneous and were dominated by the Grubii variant (95.2%), VNI (94.0%), α mating (100%), and ST5 (89.3%) genotypes. Other rare types, including ST4, 31, 278, 7, 57 and 106, were also found. Phenotypically variant and non-wild-type strains were found in Guangdong, and a significant proportion of these strains had MIC values near the ECV values towards the 6 antifungal drugs, and resistance was observed for 4 out of 6 drugs. The molecular type was highly homogeneous but compositionally diverse, with rare types found. Enhanced surveillance of the aetiology and evolution and continuous monitoring of antifungal susceptibility are needed to provide references for decision-making in the health sector and optimization of disease prevention and control.