APLpred: A machine learning-based tool for accurate prediction and characterization of asparagine peptide lyases using sequence-derived optimal features.

Asparagine peptide lyase (APL) is among the seven groups of proteases, also known as proteolytic enzymes, which are classified according to their catalytic residue. APLs are synthesized as precursors or propeptides that undergo self-cleavage through autoproteolytic reaction. At present, APLs are grouped into 10 families belonging to six different clans of proteases. Recognizing their critical roles in many biological processes including virus maturation, and virulence, accurate identification and characterization of APLs is indispensable. Experimental identification and characterization of APLs is laborious and time-consuming. Here, we developed APLpred, a novel support vector machine (SVM) based predictor that can predict APLs from the primary sequences. APLpred was developed using Boruta-based optimal features derived from seven encodings and subsequently trained using five machine learning algorithms. After evaluating each model on an independent dataset, we selected APLpred (an SVM-based model) due to its consistent performance during cross-validation and independent evaluation. We anticipate APLpred will be an effective tool for identifying APLs. This could aid in designing inhibitors against these enzymes and exploring their functions. The APLpred web server is freely available at https://procarb.org/APLpred/.

Synthesis, delineation and technological advancements of algae biochar for sustainable remediation of the emerging pollutants from wastewater-a review.

The use of algae for value-added product and biorefining applications is enchanting attention among researchers in recent years due to its remarkable photosynthetic ability, adaptability, and capacity to accumulate lipids and carbohydrates. Algae biomass, based on its low manufacturing costs, is relatively renewable, sustainable, environmentally friendly and economical in comparison with other species. High production rate of algae provides a unique opportunity for its conversion to biochar with excellent physicochemical properties, viz. high surface area and pore volume, high adsorption capacity, abundant functional groups over surface, etc. Despite several potential algal-biochar, a detailed study on its application for removal of emerging contaminants from wastewater is limited. Therefore, this technical review is being carried out to evaluate the specific elimination of inorganic and organic pollutants from wastewater, with a view to assessing adsorption performances of biochar obtained from various algae species. Species-specific adsorption of emerging pollutants from wastewater have been discussed in the present review. The promising methods like pyrolysis, gasification, dry and wet torrefaction for the production of algae biochar are highlighted. The strategies include chemical and structural modifications of algae biochar for the removal of toxic contaminants have also been considered in the current work. The overall aim of this review is to confer about the synthesis, technological advancements, delineation and application of algae biochar for the treatment of wastewater.

Computational prediction of phosphorylation sites of SARS-CoV-2 infection using feature fusion and optimization strategies.

SARS-CoV-2's global spread has instigated a critical health and economic emergency, impacting countless individuals. Understanding the virus's phosphorylation sites is vital to unravel the molecular intricacies of the infection and subsequent changes in host cellular processes. Several computational methods have been proposed to identify phosphorylation sites, typically focusing on specific residue (S/T) or Y phosphorylation sites. Unfortunately, current predictive tools perform best on these specific residues and may not extend their efficacy to other residues, emphasizing the urgent need for enhanced methodologies. In this study, we developed a novel predictor that integrated all the residues (STY) phosphorylation sites information. We extracted ten different feature descriptors, primarily derived from composition, evolutionary, and position-specific information, and assessed their discriminative power through five classifiers. Our results indicated that Light Gradient Boosting (LGB) showed superior performance, and five descriptors displayed excellent discriminative capabilities. Subsequently, we identified the top two integrated features have high discriminative capability and trained with LGB to develop the final prediction model, LGB-IPs. The proposed approach shows an excellent performance on 10-fold cross-validation with an ACC, MCC, and AUC values of 0.831, 0.662, 0.907, respectively. Notably, these performances are replicated in the independent evaluation. Consequently, our approach may provide valuable insights into the phosphorylation mechanisms in SARS-CoV-2 infection for biomedical researchers.

Improved Antibacterial Activity of Peptide Nisin with Pyrrole-Based Ionic Liquids Having Bis(trifluoromethylsulfonyl)imide as a Counterion: A Synergistic Approach to Combat Bacterial Infections.

Bacterial resistance against antimicrobial drugs is a forthcoming threat to the prevention and treatment of developing bacterial infections. Hence, the development of new antimicrobial therapy or therapeutic drugs is desperately needed. A combination of antibiotics exhibits synergistic antibacterial effects. As the combination approach of antibiotics has always shown better results against pathogens compared to monotherapy with an antibiotic, we focused on creating a new combination that may reduce the chances of strains attaining resistance, consequently lowering the toxicity factor associated with the consumption of high amounts of antibiotics. Nisin, a food preservative and potential antibiotic, shows antibacterial activity against Gram-positive strains. Since the past decade, ionic liquids (ILs) have proven to be an important class of potential antibacterial agents. In our study, we studied the effect of pyrrolidinium-based ILs and arrived at a noncovalent conjugate formed by combining nisin with ILs. The conjugates were tested against a couple of clinically relevant microorganisms, namely, Escherichia coli and Staphylococcus aureus. We reached a novel discovery that the combination of sodium/iodide symporter (NIS) and IL exhibited inhibitory effects against Gram-negative bacteria, which was not observed with NIS alone. The results showed remarkable improvement in the minimum inhibitory concentration (MIC) value of NIS in the presence of ILs targeted against both microorganisms. Further, flow cytometry and confocal microscopy results revealed the membrane disruption efficiency of the best combination obtained, leading to cell death. Additionally, the complexation of nisin and ILs was studied using various techniques, such as surface tension, dynamic light scattering, absorption spectroscopy, and molecular docking.

Thermodynamic Insights of the Molecular Interactions of Dopamine (Neurotransmitter) with Anionic Surfactant in Non-Aqueous Media.

This study was aimed at establishing the interactions prevailing in an anionic surfactant, sodium dodecyl sulfate, and dopamine hydrochloride in an alcoholic (ethanol) media by using volumetric, conductometric, and tensiometric techniques. Various methods were utilized to estimate the critical micelle concentration (cmc) values at different temperatures. The entire methods yielded the same cmc values. The corresponding thermodynamic parameters viz. the standard free energy of micellization (Gmico), enthalpy of micellization (Hmico), and entropy of micellization (Smico) were predicted by applying the pseudo-phase separation model. The experimental density data at different temperatures (298.15 K, 303.15 K, 308.15 K, and 313.15 K) were utilized to estimate the apparent molar volumes (Vϕo) at an infinite dilution, apparent molar volumes (Vφcmc) at the critical micelle concentration, and apparent molar volumes (ΔVφm) upon micellization. Various micellar and interfacial parameters, for example, the surface excess concentration (Γmax), standard Gibbs free energy of adsorption at the interface (ΔGoad), and the minimum surface area per molecule (Amin), were appraised using the surface tension data. The results were used to interpret the intermolecular interactions prevailing in the mixed systems under the specified experimental conditions.

Limosilactobacillus fermentum KAU0021 Abrogates Mono- and Polymicrobial Biofilms Formed by Candida albicans and Staphylococcus aureus.

Candida albicans and Staphylococcus aureus, representing two different kingdoms, are the most frequently isolated pathogens from invasive infections. Their pathogenic attributes, combined with drug resistance, make them a major threat and a challenge to successful treatments, mainly when involved in polymicrobial biofilm-associated infections. In the present study, we investigated the antimicrobial potential of Lactobacillus metabolite extracts (LMEs) purified from cell-free supernatant of four Lactobacillus strains (KAU007, KAU0010, KAU0021, and Pro-65). Furthermore, LME obtained from the strain KAU0021 (LMEKAU0021), being the most effective, was analyzed for its anti-biofilm property against mono- and polymicrobial biofilms formed by C. albicans and S. aureus. The impact of LMEKAU0021 on membrane integrity in single and mixed culture conditions was also evaluated using propidium iodide. The MIC values recorded for LMEKAU0021 was 406 µg/mL, 203 µg/mL, and 406 µg/mL against planktonic cells of C. albicans SC5314, S. aureus and polymicrobial culture, respectively. The LMEKAU0021 at sub-MIC values potentially abrogates both biofilm formation as well as 24 h mature mono- and polymicrobial biofilms. These results were further validated using different microscopy and viability assays. For insight mechanism, LMEKAU0021 displayed a strong impact on cell membrane integrity of both pathogens in single and mixed conditions. A hemolytic assay using horse blood cells at different concentrations of LMEKAU0021 confirmed the safety of this extract. The results from this study correlate the antimicrobial and anti-biofilm properties of lactobacilli against bacterial and fungal pathogens in different conditions. Further in vitro and in vivo studies determining these effects will support the aim of discovering an alternative strategy for combating serious polymicrobial infections caused by C. albicans and S. aureus.

Biosynthesis of Novel Ag-Cu Bimetallic Nanoparticles from Leaf Extract of Salvia officinalis and Their Antibacterial Activity.

Bimetallic nanoparticles exhibit bifunctional or synergistic effects prevailing between two metals with the capabilities of enhanced electronic, catalytic, and optical properties. Green synthetic routes have gained tremendous interest because of the noninvolvement of toxic and harmful chemical reagents in preparation. Therefore, we develop bimetallic Ag-Cu nanoparticles (Ag-Cu NPs) through an eco-friendly and biocompatible preparation method. In this study, Ag-Cu NPs have been synthesized from leaf extracts of the commonly known sage, S. officinalis. The extract has a rich phytochemical composition, including bioreducing polyphenols, flavonoids, and capping/stabilizing agents. An array of well-known spectroscopic and microscopic techniques were used to characterize the as-prepared Ag-Cu bimetallic nanoparticles, including X-ray diffraction (XRD), ultraviolet-visible spectroscopy, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy, transmission electron microscopy, and energy-dispersive X-ray spectroscopy. The size of the Ag-Cu NPs was found to be 50 nm with a spherical shape and an almost uniform distribution. The antibacterial effect was further evaluated using agar well diffusion and disc diffusion assays. Ag-Cu NPs exhibit antibacterial and antibiofilm properties against Gram-positive and Gram-negative bacteria strains. The minimum inhibitory concentration (MIC) of Ag-Cu NPs was between 5 g/mL and 15 g/mL. The Ag-Cu NPs inhibit biofilm formation at 25 g/mL and 50 g/mL. The results of biogenic Ag-Cu NPs provide novel antibacterial activity against Gram-positive and Gram-negative bacteria, as well as antibiofilm activity. Hence, Ag-Cu NPs might serve as a novel antibacterial agent with potential antibacterial and antibiofilm properties.

Facile Synthesis of Magnetic Nigella Sativa Seeds: Advances on Nano-Formulation Approaches for Delivering Antioxidants and Their Antifungal Activity against Candida albicans.

This article reports on incorporating magnetic nanoparticles into natural carbon frameworks derived from Nigella Sativa seeds and their synthesis via co-precipitation reactions for application in biomedicine. The magnetic Nigella Sativa Seeds (Magnetic NSS), a metal oxide-based bio-nanomaterial, has shown excellent water diaper presence due to the presence of a wide range of oxygenous hydroxyl and carboxyl groups. The physicochemical properties of the composites were characterized extensively using Fourier transform infrared spectroscopy (FTIR), powder-X-ray diffraction (XRD), scanning electron microscopy (SEM), elemental analysis, transmission electron microscopy (TEM), and vibrating-sample magnetometer. Furthermore, synthesized magnetic NSS showed antioxidant and antifungal activity. The antifungal susceptibility was further tested against Candida albicans with a MIC value of 3.125 µg/mL. Analysis of antioxidant defense enzymes was determined quantitatively; the results suggested that antioxidant enzyme activity increase with increased magnetic NSS concentration. Furthermore, biofilm inhibition assay from scanning electron microscopy results revealed that magnetic NSS at the concentration of 3.5 µg/mL has anti-biofilm properties and can disrupt membrane integrity.

Lactiplantibacillus plantarum KAU007 Extract Modulates Critical Virulence Attributes and Biofilm Formation in Sinusitis Causing Streptococcus pyogenes.

Streptococcus pyogenes is one of the most common bacteria causing sinusitis in children and adult patients. Probiotics are known to cause antagonistic effects on S. pyogenes growth and biofilm formation. In the present study, we demonstrated the anti-biofilm and anti-virulence properties of Lactiplantibacillus plantarum KAU007 against S. pyogenes ATCC 8668. The antibacterial potential of L. plantarum KAU007 metabolite extract (LME) purified from the cell-free supernatant of L. plantarum KAU007 was evaluated in terms of minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC). LME was further analyzed for its anti-biofilm potential using crystal violet assay and microscopic examination. Furthermore, the effect of LME was tested on the important virulence attributes of S. pyogenes, such as secreted protease production, hemolysis, extracellular polymeric substance production, and cell surface hydrophobicity. Additionally, the impact of LME on the expression of genes associated with biofilm formation and virulence attributes was analyzed using qPCR. The results revealed that LME significantly inhibited the growth and survival of S. pyogenes at a low concentration (MIC, 9.76 µg/mL; MBC, 39.06 µg/mL). Furthermore, LME inhibited biofilm formation and mitigated the production of extracellular polymeric substance at a concentration of 4.88 µg/mL in S. pyogenes. The results obtained from qPCR and biochemical assays advocated that LME suppresses the expression of various critical virulence-associated genes, which correspondingly affect various pathogenicity markers and were responsible for the impairment of virulence and biofilm formation in S. pyogenes. The non-hemolytic nature of LME and its anti-biofilm and anti-virulence properties against S. pyogenes invoke further investigation to study the role of LME as an antibacterial agent to combat streptococcal infections.

The Inhibition of SARS-CoV-2 and the Modulation of Inflammatory Responses by the Extract of Lactobacillus sakei Probio65.

In the three years since the first outbreak of COVID-19 in 2019, the SARS-CoV-2 virus has continued to be prevalent in our community. It is believed that the virus will remain present, and be transmitted at a predictable rate, turning endemic. A major challenge that leads to this is the constant yet rapid mutation of the virus, which has rendered vaccination and current treatments less effective. In this study, the Lactobacillus sakei Probio65 extract (P65-CFS) was tested for its safety and efficacy in inhibiting SARS-CoV-2 replication. Viral load quantification by RT-PCR showed that the P65-CFS inhibited SARS-CoV-2 replication in human embryonic kidney (HEK) 293 cells in a dose-dependent manner, with 150 mg/mL being the most effective concentration (60.16% replication inhibition) (p < 0.05). No cytotoxicity was inflicted on the HEK 293 cells, human corneal epithelial (HCE) cells, or human cervical (HeLa) cells, as confirmed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The P65-CFS (150 mg/mL) also reduced 83.40% of reactive oxidizing species (ROS) and extracellular signal-regulated kinases (ERK) phosphorylation in virus-infected cells, both of which function as important biomarkers for the pathogenesis of SARS-CoV-2. Furthermore, inflammatory markers, including interferon-α (IFN-α), IFN-ß, and interleukin-6 (IL-6), were all downregulated by P65-CFS in virus-infected cells as compared to the untreated control (p < 0.05). It was conclusively found that L. sakei Probio65 showed notable therapeutic efficacy in vitro by controlling not only viral multiplication but also pathogenicity; this finding suggests its potential to prevent severe COVID-19 and shorten the duration of infectiousness, thus proving useful as an adjuvant along with the currently available treatments.

Characterization of Defensin-like Protein 1 for Its Anti-Biofilm and Anti-Virulence Properties for the Development of Novel Antifungal Drug against Candida auris.

Candida auris has emerged as a pan-resistant pathogenic yeast among immunocompromised patients worldwide. As this pathogen is involved in biofilm-associated infections with serious medical manifestations due to the collective expression of pathogenic attributes and factors associated with drug resistance, successful treatment becomes a major concern. In the present study, we investigated the candidicidal activity of a plant defensin peptide named defensin-like protein 1 (D-lp1) against twenty-five clinical strains of C. auris. Furthermore, following the standard protocols, the D-lp1 was analyzed for its anti-biofilm and anti-virulence properties. The impact of these peptides on membrane integrity was also evaluated. For cytotoxicity determination, a hemolytic assay was conducted using horse blood. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values ranged from 0.047-0.78 mg/mL and 0.095-1.56 mg/mL, respectively. D-lp1 at sub-inhibitory concentrations potentially abrogated both biofilm formation and 24-h mature biofilms. Similarly, the peptide severely impacted virulence attributes in the clinical strain of C. auris. For the insight mechanism, D-lp1 displayed a strong impact on the cell membrane integrity of the test pathogen. It is important to note that D-lp1 at sub-inhibitory concentrations displayed minimal hemolytic activity against horse blood cells. Therefore, it is highly useful to correlate the anti-Candida property of D-lp1 along with anti-biofilm and anti-virulent properties against C. auris, with the aim of discovering an alternative strategy for combating serious biofilm-associated infections caused by C. auris.

Evaluation of Lactiplantibacillus plantarum KAU007 against Low-Pathogenic Avian Influenza Virus (H9N2).

Avian influenza A viruses (AIVs) pose a persistent threat to humans owing to their reassortment and antigenic drift properties. Among them is H9N2, a low-pathogenic avian influenza virus first discovered in the non-human host and later found infective to humans with huge pandemic potential. In recent years, antiviral resistance has become an increasing threat to public health. Additionally, vaccination against AIVs is becoming increasingly challenging with little success due to antigenic drift. This has resulted in a growing demand for products that can replace the presently in-use medications and the development of innovative antiviral therapies. In this study, we systematically investigate the antiviral potential of lactic acid bacteria against H9N2. Bacteria that produce lactic acid are commonly used in food processing. In addition, these bacteria are considered more affordable, effective, and safe "nutraceuticals" than other alternative medicines. We tested Lactiplantibacillus plantarum KAU007 against the low-pathogenic avian influenza virus (H9N2). As confirmed by the hemagglutination assay, KAU007 showed potent antiviral activity against H9N2 and vigorous antioxidant activity. The CFCS showed a dose-dependent reduction in the levels of IL-6 and IFN-γ. Thus, KAU007 might be considered a potential H9N2 target-based probiotic.

Characterization of the Secretome of Pathogenic Candida glabrata and Their Effectiveness against Systemic Candidiasis in BALB/c Mice for Vaccine Development.

Infections by non-albicans Candida species have increased drastically in the past few decades. Candida glabrata is one of the most common opportunistic fungal pathogens in immunocompromised individuals, owing to its capability to attach to various human cell types and medical devices and being intrinsically weakly susceptible to azoles. Immunotherapy, including the development of antifungal vaccines, has been recognized as an alternative approach for preventing and treating fungal infections. Secretory proteins play a crucial role in establishing host-pathogen interactions and are also responsible for eliciting an immune response in the host during candidiasis. Therefore, fungal secretomes can provide promising protein candidates for antifungal vaccine development. This study attempts to uncover the presence of immunodominant antigenic proteins in the C. glabrata secretome and delineate their role in various biological processes and their potency in the development of antifungal vaccines. LC-MS/MS results uncovered that C. glabrata secretome consisted of 583 proteins, among which 33 were identified as antigenic proteins. The protection ability of secretory proteins against hematogenously disseminated infection caused by C. glabrata was evaluated in BALB/c mice. After immunization and booster doses, all the animals were challenged with a lethal dose of C. glabrata. All the mice showing signs of distress were sacrificed post-infection, and target organs were collected, followed by histopathology and C. glabrata (CFU/mg) estimation. Our results showed a lower fungal burden in target organs and increased survival in immunized mice compared to the infection control group, thus revealing the immunogenic property of secreted proteins. Thus, identified secretome proteins of C. glabrata have the potential to act as antigenic proteins, which can serve as potential candidates for the development of antifungal vaccines. This study also emphasizes the importance of a mass-spectrometry approach to identifying the antigenic proteins in C. glabrata secretome.

Human ß defensins-1, an antimicrobial peptide, kills Candida glabrata by generating oxidative stress and arresting the cell cycle in G0/G1 phase.

Candida glabrata is the most frequently isolated non-albicans Candida species in clinical samples and is known to develop resistance to commonly used antifungal drugs. Human ß defensins (hBDs) are antimicrobial peptides of immune systems and are active against a broad range of pathogens including Candida species. Herein, the antifungal effect of hBD-1 and its mechanism of action in C. glabrata was studied. The antifungal susceptibility of hBD-1 against C. glabrata was calculated by broth microdilution assay. To study the mechanism of antifungal action, the impact of hBD-1 on cell cycle, expression of oxidative stress enzymes, and membrane disintegration were assessed. The susceptibility results confirmed that hBD-1 possessed the minimum inhibitory concentration of 3.12 µg/mL and prevented the growth and caused yeast cell death to various extents. The peptide at subinhibitory and inhibitory concentrations blocked the cell cycle in C. glabrata in G0/G1 phase and disturbed the activity of primary and secondary antioxidant enzymes. Furthermore, at higher concentrations disruption of membrane integrity was observed. Altogether, hBD-1 showed candidicidal activity against C. glabrata and was able to induce oxidative stress and arrested cell cycle in C. auris and therefore has a potential to be developed as an antifungal drug against C. glabrata.

Combination Effect of Novel Bimetallic Ag-Ni Nanoparticles with Fluconazole against Candida albicans.

The increasing frequency of antifungal drug resistance among pathogenic yeast "Candida" has posed an immense global threat to the public healthcare sector. The most notable species of Candida causing most fungal infections is Candida albicans. Furthermore, recent research has revealed that transition and noble metal combinations can have synergistic antimicrobial effects. Therefore, a one-pot seedless biogenic synthesis of Ag-Ni bimetallic nanoparticles (Ag-Ni NPs) using Salvia officinalis aqueous leaf extract is described. Various techniques, such as UV-vis, FTIR, XRD, SEM, EDX, and TGA, were used to validate the production of Ag-Ni NPs. The antifungal susceptibility of Ag-Ni NPs alone and in combination with fluconazole (FLZ) was tested against FLZ-resistant C. albicans isolate. Furthermore, the impacts of these NPs on membrane integrity, drug efflux pumps, and biofilms formation were evaluated. The MIC (1.56 µg/mL) and MFC (3.12 µg/mL) results indicated potent antifungal activity of Ag-Ni NPs against FLZ-resistant C. albicans. Upon combination, synergistic interaction was observed between Ag-Ni NPs and FLZ against C. albicans 5112 with a fractional inhibitory concentration index (FICI) value of 0.31. In-depth studies revealed that Ag-Ni NPs at higher concentrations (3.12 µg/mL) have anti-biofilm properties and disrupt membrane integrity, as demonstrated by scanning electron microscopy results. In comparison, morphological transition was halted at lower concentrations (0.78 µg/mL). From the results of efflux pump assay using rhodamine 6G (R6G), it was evident that Ag-Ni NPs blocks the efflux pumps in the FLZ-resistant C. albicans 5112. Targeting biofilms and efflux pumps using novel drugs will be an alternate approach for combatting the threat of multi-drug resistant (MDR) stains of C. albicans. Therefore, this study supports the usage of Ag-Ni NPs to avert infections caused by drug resistant strains of C. albicans.

Polyphenol-Capped Biogenic Synthesis of Noble Metallic Silver Nanoparticles for Antifungal Activity against Candida auris.

In terms of reduced toxicity, the biologically inspired green synthesis of nanoparticles has emerged as a promising alternative to chemically fabricated nanoparticles. The use of a highly stable, biocompatible, and environmentally friendly aqueous extract of Cynara cardunculus as a reducing and capping agent in this study demonstrated the possibility of green manufacturing of silver nanoparticles (CC-AgNPs). UV-visible spectroscopy validated the development of CC-AgNPs, indicating the surface plasmon resonance (SPR) λmax band at 438 nm. The band gap of CC-AgNPs was found to be 2.26 eV. SEM and TEM analysis examined the surface morphology of CC-AgNPs, and micrographs revealed that the nanoparticles were spherical. The crystallinity, crystallite size, and phase purity of as-prepared nanoparticles were confirmed using XRD analysis, and it was confirmed that the CC-AgNPs were a face-centered cubic (fcc) crystalline-structured material. Furthermore, the role of active functional groups involved in the reduction and surface capping of CC-AgNPs was revealed using the Fourier transform infrared (FTIR) spectroscopic technique. CC-AgNPs were mostly spherical and monodispersed, with an average size of 26.89 nm, and were shown to be stable for a longer period without any noticeable change at room temperature. Further, we checked the antifungal mechanism of CC-AgNPs against C. auris MRL6057. The minimum inhibitory concentrations (MIC) and minimum fungicidal concentrations (MFC) were 50.0 µg/mL and 100.0 µg/mL respectively. The cell count and viability assay confirmed the fungicidal potential of CC-AgNPs. Further, the analysis showed that CC-AgNPs could induce apoptosis and G2/M phase cell cycle arrest in C. auris MRL6057. Our results also suggest that the CC-AgNPs were responsible for the induction of mitochondrial toxicity. TUNEL assay results revealed that higher concentrations of CC-AgNPs could cause DNA fragmentation. Therefore, the present study suggested that CC-AgNPs hold the capacity for antifungal drug development against C. auris infections.

Lornoxicam-Loaded Chitosan-Decorated Nanoemulsion: Preparation and In Vitro Evaluation for Enhanced Transdermal Delivery.

Nanoemulsions are promising drug delivery systems for the administration of poorly soluble drugs like lornoxicam (LRX) by oral or parenteral routes. Such formulations work perfectly for transdermal delivery of lornoxicam-type drugs. It has also been established that formulating such a delivery system is highly dependent on the presence, type, and concentration of excipients taking part in the formulation. The inherent characteristics of nanoemulsion (NE), i.e., smaller globule size and excipient nature, facilitate the drug's passage through skin. The current study was aimed at the development of an NE-based formulation of LRX to improve the drug solubility in vitro as well as to enhance drug skin permeation to promote therapeutic outcome in appropriate time. Spontaneous self-emulsification technique was utilized to develop optimized LRX-encapsulated NE-based formulations. ATR-FTIR spectra of the pure drug and various formulations did not show any interaction between the drug and various formulation excipients showing compatibility. Globule size for stable formulations ranged between 63-168 nm. These formulations were characterized for viscosity, surface tension, pH, drug encapsulation efficiency, in vitro drug release, and drug skin permeation studies. Chitosan-decorated optimized NE formulation of LRX showed about 58.82% cumulative drug release, showing an anomalous non-Fickian diffusion mechanism of drug release. Drug encapsulation efficiency, in vitro drug release, and skin permeation studies exhibited promising results. An appreciable drug entrapment efficiency was exhibited by optimized NE formulations LRX-6, 71.91 ± 3.17% and C-LRX, 65.25 ± 4.89%. Permeability parameters like enhancement ratio (Er), permeability constant (Kp), and steady state flux (Jss) showed higher values and exhibited good results based on formulation type. The selected promising formulation type "LRX-6" showed significantly different results as compared to other formulations (LRX-4, 5, and 7). The skin permeation property of the LRX-6 formulation was compared to similar chitosan-based formulations and was found to have better skin permeation results than chitosan-based formulations. This study clearly exhibited that an LRX-containing NE-based formulation can be formulated to form a stable drug delivery system. Such formulations are promising in terms of physicochemical characteristics, improved solubility, and high skin permeation potential.

Potential Antiviral Activity of Lactiplantibacillus plantarum KAU007 against Influenza Virus H1N1.

The development of antiviral resistance has exacerbated a growing threat to public health. As a result, there is increasing demand for unconventional antivirals that can effectively replace the presently in-use drugs. Lactic acid-producing bacteria (LAB) are among the most common bacteria used in the food industry. These bacteria play an essential role in the fermentation of many foods and feed. Additionally, these bacteria are considered more economical, efficient, and safe "nutraceuticals" in the health care arsenal. Therefore, we carried out the screening and molecular characterization of raw camel milk LAB isolates and tested their inhibitory activity against influenza virus H1N1. The strain that exhibited the highest antiviral activity against the H1N1 virus, confirmed by hemagglutination assay, was identified as Lactiplantibacillus plantarum KAU007. The study also confirmed the non-cytotoxic behavior of CFCS isolated from KAU007 against MDCK cells, approving its safety concern against the mammalian cells. Besides, CFCS at 5 and 10 mg/mL significantly decreased the level of IFN-γ (p < 0.001 and p < 0.001) and IL-6 (p < 0.001 and p < 0.005) in a dose-dependent manner, respectively. This is a preliminary report about the anti-influenza activity of KAU007 isolated from camel milk. This study reinforces that camel milk contains beneficial LAB isolates with antagonistic properties against the H1N1 influenza virus.

Phytochemical Screening of Rosmarinus officinalis L. as a Potential Anticholinesterase and Antioxidant-Medicinal Plant for Cognitive Decline Disorders.

The inhibition of acetylcholinesterase (AChE) by cholinergic agents has been promoted as a potent strategy for treating and managing cognitive decline disorders. A wide range of natural products has long been used as potential sources or formulations of cholinergic inhibitors. Therefore, this study aimed to evaluate different Rosmarinus officinalis L. (R. officinalis) extracts for their AChE inhibitory activity using galanthamine as a standard AChE inhibitor. In this study, the ethyl-acetate extract (at a concentration of 250 µg/mL) exhibited the greatest inhibitory effect against AChE with significant inhibition of 75%, comparable to the inhibitor galanthamine with an inhibition of 88%. Kinetic analysis revealed that the extracts could induce a mixed type of inhibition, as observed in the case of galanthamine, with the highest increased Km and decreased Vmax values in the ethyl acetate extract. The antioxidant potential of the three extracts tested was found to be in the order of ethyl-acetate > ethanol > aqueous, with IC50 values of 272 µg/mL, 387 µg/mL, and 534 µg/mL, respectively. Ethyl-acetate was found to have the highest total phenolic content in all extracts. Further, in silico study showed structural binding characterization of rosmarinic acid and carnosic acid with human AChE enzyme. Rosmarinic acid showed strong binding and formed two hydrogen-bonding interactions with Ser-293 and Arg-296. In light of this, the ethyl-acetate extract of the plant may provide some novel potential pharmacological leads for treating and managing cognitive disorders such as Alzheimer's.

A Bioinformatics Approach to Identifying Potential Biomarkers for Cryptosporidium parvum: A Coccidian Parasite Associated with Fetal Diarrhea.

Cryptosporidium parvum (C. parvum) is a protozoan parasite known for cryptosporidiosis in pre-weaned calves. Animals and patients with immunosuppression are at risk of developing the disease, which can cause potentially fatal diarrhoea. The present study aimed to construct a network biology framework based on the differentially expressed genes (DEGs) of C. parvum infected subjects. In this way, the gene expression profiling analysis of C. parvum infected individuals can give us a snapshot of actively expressed genes and transcripts under infection conditions. In the present study, we have analyzed microarray data sets and compared the gene expression profiles of the patients with the different data sets of the healthy control. Using a network medicine approach to identify the most influential genes in the gene interaction network, we uncovered essential genes and pathways related to C. parvum infection. We identified 164 differentially expressed genes (109 up- and 54 down-regulated DEGs) and allocated them to pathway and gene set enrichment analysis. The results underpin the identification of seven significant hub genes with high centrality values: ISG15, MX1, IFI44L, STAT1, IFIT1, OAS1, IFIT3, RSAD2, IFITM1, and IFI44. These genes are associated with diverse biological processes not limited to host interaction, type 1 interferon production, or response to IL-gamma. Furthermore, four genes (IFI44, IFIT3, IFITM1, and MX1) were also discovered to be involved in innate immunity, inflammation, apoptosis, phosphorylation, cell proliferation, and cell signaling. In conclusion, these results reinforce the development and implementation of tools based on gene profiles to identify and treat Cryptosporidium parvum-related diseases at an early stage.