This study was designed to uncover the therapeutic potential of Ipomoea pes-caprae extract (IPE) and its green-synthesized selenium nanoparticles (Se NPs) in enhancing the healing of Pseudomonas aeruginosa-infected wounds in rats, with a focus on the modulation of ferroptosis and the Nrf2/HO-1 signaling pathway. The HPLC analysis of IPE revealed 14 phenolic acids and flavonoid compounds. The green-synthesized Se NPs were characterized using UV-Vis spectroscopy with peaks at 233 and 277 nm, confirming nanoparticle formation. The XRD indicated a nanocrystalline structure with an average crystallite size of 68.43 nm. The FTIR identified functional groups from the IPE involved in capping and stabilizing the Se NPs. The SEM and TEM images showed predominantly spherical particles, while EDX confirmed elemental selenium alongside carbon and oxygen, indicating phytochemical-mediated synthesis. The DLS measured a hydrodynamic size of 273.76 +/- 0.31 nm with a low polydispersity index (0.202 +/- 0.014). Also, the zeta potential analysis showed a value of -26.42 +/- 0.35 mV, suggesting strong colloidal stability. These results validate the successful green synthesis of stable and nanoscale Se NPs using IPE. An in vivo infected wound model was triggered in rats. Colorimetric, ELISA, and qRT-PCR methods to measure different biochemical markers. It was found that the Se NPs upregulated the expression of GPX4, ferritin, and HO-1, while downregulating PTGS2 and ACSL4, indicating effective suppression of ferroptosis. Immunohistochemical analysis demonstrated enhanced Nrf2 and Ki-67 expression in the Se NPs-treated group, suggesting improved antioxidant activity and cellular proliferation. A computational network pharmacology analysis was also performed, and it revealed that the phytochemicals in IPE considerably enhanced wound healing, lessened oxidative stress, and modulated ferroptosis-related markers. In conclusion, the biosynthesized Se NPs exhibited superior wound healing potential via its antioxidant, anti-inflammatory, antimicrobial, and anti-ferroptotic mechanisms.

ETHNOPHARMACOLOGICAL RELEVANCE:Cleome droserifolia (CD) has been employed traditionally by the Egyptians inhabiting desert for hyperglycemia treatment. AIM OF THE STUDY:We aimed to evaluate the capability of CD to lessen the detrimental consequences of cisplatin (CP) on testis. METHODS:CD phytoconstituents were studied by LC-ESI-MS/MS. The in vivo study involved four groups of forty male rats. Group I had a normal saline intraperitoneal (IP) injection daily; groups II, III, and IV had a single IP injection of 7 mg/kg of CP. Groups III and IV were orally administered 100 and 200 mg/kg of CD, respectively. They were treated daily for 10 days, and a single dose of IP injection of CP was taken on the third day. RESULTS:The LC-ESI-MS/MS exposed presence of 33 different compounds in CD. Also, CD at 200 mg/kg considerably reestablished the testicular weight loss and serum testosterone in CP-treated rats. Treatment with CD improved the antioxidant capabilities via downregulating malondialdehyde and increasing catalase and glutathione peroxidase activity as well as nuclear factor erythroid 2-related factor 2 immunostaining. It exerted a significant anti-inflammatory action via its decreasing effect on nuclear factor kappa beta immunostaining, and testicular level of tumor necrosis factor-α. Moreover, it exhibited an anti-apoptotic effect by downregulation of p53 and caspase-3, besides enhancing the B-cell lymphoma 2 immunostaining. In addition, molecular docking studies revealed that the major phytochemicals in CD had favorable binding affinities to the key proteins of apoptosis. CONCLUSION:CD demonstrated promising antioxidant, anti-inflammatory, and anti-apoptotic properties.

The rise of difficult-to-treat fungal infections necessitates novel therapeutic strategies. In this study, endophytic fungi were isolated from Acalypha hispida leaves and molecularly identified as Penicillium oxalicum via 18S rRNA sequencing. LC–MS/MS analysis of the fungal extract revealed major bioactive compounds, including linoleic acid, sinapinic acid, alternariol monomethyl ether, ellagic acid, and kaurenic acid. Oily-core poly (ethylene glycol) methyl ether-block-poly(lactide-co-glycolide) nanocapsules (PEGylated PLGA NCs) were developed to encapsulate the fungal extract, improving stability and bioavailability. The PEGylated PLGA NCs exhibited controlled particle size, positive surface charge, and spherical morphology. In vitro, the PEGylated PLGA NCs demonstrated antifungal activity against Candida albicans with inhibition zones of 10–14 mm. In vivo, treatment significantly improved histological features of the kidney, liver, and spleen, and reduced tumor necrosis factor-alpha and cyclooxygenase-2 expression. In silico studies further confirmed the potential of the major compounds of the fungal extract to inhibit C. albicans aspartic proteinases SAP4-6. These findings suggest that PEGylated PLGA NCs loaded with P. oxalicum extract represent a promising antifungal therapeutic strategy.

The search for antibacterial agents remains a global priority, particularly against multidrug-resistant pathogens. In this study, microbial transformation of 1,8-dihydroxyanthraquinone (chrysazin) was performed to obtain derivatives with enhanced antibacterial activity. Biotransformation using Absidia corymbifera AUMC 7104 and Beauveria bassiana AUMC 5133 yielded two metabolites: 1,2,5,8-tetrahydroxyanthraquinone (1, quinalizarin) and 1,8-dihydroxy-3-methyl-anthraquinone (2, chrysophanol), respectively. Their structures were elucidated by HRESIMS, 1D/2D NMR analyses. To the best of our knowledge, this is the first report of quinalizarin production through microbial biotransformation. The antibacterial activities of both derivatives (1 and 2), along with their parent compound, were evaluated against multidrug-resistant Acinetobacter baumannii through in vitro assays. Compound 1 exhibited the least MIC values of 512–1024 µg/mL against A. baumannii isolates and was selected for further in vivo and in silico studies. In the murine infection model, the compound 1-treated group (group IV) exhibited a significant reduction (p < 0.05) in bacterial burden compared to the positive control group (group II). Immunohistochemical analysis of kidney, liver, and spleen tissues revealed modulation of pro-inflammatory cytokines, including IL-1β, IL-6, and TNF-α, indicating anti-inflammatory properties alongside antibacterial activity. Molecular docking suggests a potential multi-target mechanism for compound 1, with favorable interactions with five critical bacterial proteins: MetRS, MurB, LptG/F, DXR, and MacB efflux transporter. These findings highlight quinalizarin as a potential lead compound for further antibacterial development. • Fungal biotransformation of chrysazin yielded the antibacterial agent quinalizarin. • Quinalizarin revealed in vitro and in vivo actions against Acinetobacter baumannii. • Docking studies suggested favorable interactions with multiple bacterial targets.

Respiratory infections remain a leading cause of morbidity and mortality, necessitating new therapeutic strategies. This study evaluated the antiviral and antibacterial activities of harmine, harmaline, and harmalacidine hydrochloride against H1N1 influenza virus and Staphylococcus aureus, key respiratory pathogens. The in vitro antiviral activity of the tested compounds against the H1N1 virus was evaluated using a plaque assay. Harmalacidine hydrochloride demonstrated notable activity, with an IC50 of 68.2 ± 0.8 µg/mL, while harmine and harmaline showed no significant effects at non-cytotoxic concentrations. The potential antibacterial action of the tested compounds was initially investigated by agar well diffusion method, which revealed clear zones of inhibition around the wells. Subsequently, their minimum inhibitory concentrations (MICs) were recorded using the broth microdilution method. Harmalacidine hydrochloride exhibited the highest antibacterial action with MICs from 16 to 128 µg/mL. Based on these findings, further investigations were conducted to assess the effect of harmalacidine hydrochloride on membrane integrity and permeability, cellular morphology, and biofilm formation. A noticeable reduction (p < 0.05) in the membrane integrity and a distinct escalation (p < 0.05) in the permeability were noticed in 46.15% and 53.85% of the tested isolates, respectively. Moreover, scanning electron microscopy revealed pronounced distortion in cellular morphology following harmalacidine hydrochloride treatment. The compound also exhibited antibiofilm activity, as demonstrated by the crystal violet assay, alongside a downregulation of biofilm-associated gene expression. Molecular docking revealed that harmalacidinium ion binds strongly to the Accessory Gene Regulator A (AgrA) of S. aureus, suggesting antibacterial activity through inhibition of quorum sensing-mediated virulence. It also showed high affinity for H1N1 neuraminidase and polymerase basic protein 2 (PB2), indicating potential antiviral activity. However, experimental enzyme assays and in vivo studies are required to confirm the proposed antiviral and antibacterial mechanisms.

Wound care is still a global concern due to the widespread occurrence of bacterial infections, particularly those caused by Pseudomonas aeruginosa (PA), a pathogen known for its intrinsic resistance to multiple antibiotics. This growing resistance underscores the urgent need for alternative therapeutic strategies. Folk medicine has long utilized Zygophyllum simplex L., a traditional medicinal plant, to treat eye infections, inflammation, and a variety of skin conditions, including rough and hardened patches. Nevertheless, there is currently little scientific proof of its dermatological effectiveness, despite its ethnopharmacological significance. In this study, the n-hexane extract of Z. simplex was saponified to yield saponifiable (Sap-ZSHE) and unsaponifiable (Unsap-ZSHE) fractions, which were subsequently analyzed by GC/MS to determine their chemical composition. The antibacterial and wound-healing activities of these lipoidal fractions were evaluated using both in vitro and in vivo models of PA-infected wounds. GC/MS analysis revealed 21 compounds in the Sap-ZSHE, including unsaturated (47 %) and saturated (53 %) fatty acids or their methyl esters and 25 compounds in the Unsap-ZSHE, including a fatty aldehyde (44.91 %), triterpenes (30.9 %), diterpenes (11.61 %), and sterols (8.46 %) as the predominant compounds. The Sap-ZSHE fraction demonstrated the strongest antibacterial activity, with minimum inhibitory concentrations (MICs) of 32–64 µg/mL against PA isolates, significantly surpassing the total n-hexane extract and Unsap-ZSHE (MICs: 256–1024 µg/mL). In vivo evaluation using a PA-infected burn wound model in rats revealed that Sap-ZSHE markedly improved wound healing, as evidenced by reduced bacterial load, accelerated wound closure, enhanced collagen deposition, and normalized histoarchitecture. Immunohistochemical analysis confirmed a reduction in inflammatory markers (TNF-α, IL-6, NF-κB), while qRT-PCR showed upregulation of the tissue repair markers PDGF and fibronectin. These findings demonstrated the therapeutic potential of Z. simplex, particularly its Sap-ZSHE fraction, in treating PA-infected wounds, validating its traditional use and supporting its development as a plant-based alternative for resistant wound infections.

Ulcerative colitis (UC) is an inflammatory condition of the intestine, resulting from an increase in oxidative stress and pro-inflammatory mediators. In this study, the extract of endophytic bacterium Rhizobium aegyptiacum was prepared for the first time using liquid chromatography-mass spectrometry (LC-MS). In addition, also for the first time, the protective potential of R. aegyptiacum was revealed using an in vivo rat model of UC. The animals were grouped into four categories: normal control (group I), R. aegyptiacum (group II), acetic acid (AA)-induced UC (group III), and R. aegyptiacum-treated AA-induced UC (group IV). In group IV, R. aegyptiacum was administered at 0.2 mg/kg daily for one week before and two weeks after the induction of UC. After sacrificing the rats on the last day of the experiment, colon tissues were collected and subjected to histological, immunohistochemical, and biochemical investigations. There was a remarkable improvement in the histological findings of the colon tissues in group IV, as revealed by hematoxylin and eosin (H E) staining, Masson’s trichrome staining, and periodic acid-Schiff (PAS) staining. Normal mucosal surfaces covered with a straight, intact, and thin brush border were revealed. Goblet cells appeared magenta in color, and there was a significant decrease in the distribution of collagen fibers in the mucosa and submucosal connective tissues. All these findings were comparable to the respective characteristics of the control group. Regarding cyclooxygenase-2 (COX-2) immunostaining, a weak immune reaction was shown in most cells. Moreover, the colon tissues were examined using a scanning electron microscope, which confirmed the results of histological assessment. A regular polygonal unit pattern was seen with crypt orifices of different sizes and numerous goblet cells. Furthermore, the levels of catalase (CAT), myeloperoxidase (MPO), nitric oxide (NO), interleukin-6 (IL-6), and interlukin-1β (IL-1β) were determined in the colonic tissues of the different groups using colorimetric assay and enzyme-linked immunosorbent assay (ELISA). In comparison with group III, group IV exhibited a significant rise (P<0.05) in the CAT level but a substantial decline (P<0.05) in the NO, MPO, and inflammatory cytokine (IL-6 and IL-1 β) levels. Based on reverse transcription-quantitative polymerase chain reaction (RT-qPCR), the tumor necrosis factor-α (TNF-α) gene expression was upregulated in group III, which was significantly downregulated (P<0.05) by treatment with R. aegyptiacum in group IV. On the contrary, the heme oxygenase-1 (HO-1) gene was substantially upregulated in group IV. Our findings imply that the oral consumption of R. aegyptiacum ameliorates AA-induced UC in rats by restoring and reestablishing the mucosal integrity, in addition to its anti-oxidant and anti-inflammatory effects. Accordingly, R. aegyptiacum is potentially effective and beneficial in human UC therapy, which needs to be further investigated in future work.

Background: Klebsiella pneumoniae is an opportunistic pathogen that frequently causes nosocomial infections and contributes to significant morbidity and mortality. Numerous factors are believed to be associated with the colonization of K. pneumoniae in hospital and community settings. K. pneumoniae frequently colonizes hospitalized individuals, leading to extraintestinal diseases like bloodstream infections (septicemia), urinary tract infections, and pneumonia. Patients in intensive care units (ICUs) are especially vulnerable to such infections. Infections with K. pneumoniae are particularly problematic in the healthcare setting for newborns, elderly, and people with impaired immune systems. Due to the severity of the diseases, resistance to numerous antibiotics, and difficulty of treatment, K. pneumoniae has drawn the interest of researchers worldwide in recent years. K. pneumoniae is a significant global antibiotic resistance source and transmitter. The global rise in resistance highlights the need for novel therapeutic options. The choice of an appropriate antibiotic for hospital-acquired infections is becoming a rising global issue due to this resistance. In this review, we reveal that K. pneumoniae is a major threat to patients in intensive care units and we discuss the mechanisms of drug resistance in K. pneumoniae. We demonstrate the mechanisms and effects of emerging novel therapeutic strategies for K. pneumoniae in recent years to overcome drug resistance in the treatment of K. pneumoniae infections for better future.

The universal spread of bacterial infections poses a significant issue in clinical settings. Pseudomonas aeruginosa is among the most commonly detected bacteria in various infections. Besides its resistance to multiple antibiotics, it also exhibits numerous virulence factors that hinder antibiotic treatment. Therefore, we intended to inspect the antibacterial and antivirulence consequences of Acacia saligna plant extract, along with phytochemical analysis of its active components by HPLC. The HPLC analysis identified 15 phenolic acids and flavonoid compounds. The A. saligna ethanol extract (ASE) demonstrated antibacterial activity, with minimum inhibitory concentrations (MICs) ranging from 128 to 1024 μg/mL. Additionally, ASE showed antibiofilm activity, reducing the percentage of vigorous and moderate biofilm-forming isolates from 78.57 % to 21.43 % as measured by the crystal violet assay. A significant downregulation of the QS genes in six out of eight P. aeruginosa isolates, with relative gene expression reduced to 0.1-0.4 for lasI, lasR, and rhlI genes, and 0.3-0.4 for rhlR gene compared to untreated controls. Furthermore, the in vivo antibacterial effects of ASE were demonstrated in P. aeruginosa-infected mice. There was a significant (p < 0.05) reduction in bacterial load in the kidneys, liver, and spleen of the ASE-treated group. Histopathological analysis revealed notable improvements in the kidney, liver, and spleen tissues in the ASE-treated group compared with the positive control. Immunohistochemical examination showed a lessening in the number of inflammatory marker-positive cells (TNF-α and IL-6). Additionally, there was a significant reduction (p < 0.05) in nitric oxide and malondialdehyde in the ASE-treated group. Therefore, ASE could be a promising antibacterial and antivirulence agent for treating P. aeruginosa infections.

Toxoplasma gondii, a protozoan parasite found in water sources, causes toxoplasmosis, with no current protocols for inactivating its oocysts in water. Staphylococcus aureus, a significant bacterial pathogen, is known for causing various illnesses, including skin infections and biofilm-related diseases. This study investigated the antibacterial and antiparasitic properties of Ipomoea palmata leaf extract, rich in phenolics, against T. gondii tachyzoites and S. aureus. I. palmata extract significantly reduced tachyzoites count in peritoneal fluids and liver smears of infected mice with alleviation of toxoplasmosis-induced hepatitis. SEM showed surface irregularities in tachyzoites from treated groups. The extract demonstrated antibacterial action against S. aureus with a minimum inhibitory concentration of 128 to 512 µg/mL, reduced biofilm formation from 69.23% to 15.38% of tested isolates, and downregulated biofilm genes (cna, fnbA, and ica) in 53.85% of isolates. Treatment with I. palmata extract improved liver architecture, reduced inflammation, and eliminated blood vessel congestion. The main phenolic acids identified by HPLC/UV analysis were chlorogenic acid, gallic acid, ellagic acid, and methyl gallate, while the predominant flavonoids were apigenin, quercetin, and naringenin. These findings highlight the potential of I. palmata extract as a natural antimicrobial and antiparasitic agent, warranting further research to isolate and evaluate its active compounds.

Loquat (Eriobotrya japonica) is a perennial tree, commonly grown as an ornamental plant and is known for its sweet edible fruits. Considering the traditional uses of medicinal plants, the aim of this study is the evaluation of the antibacterial action of isolated compounds from Eriobotrya japonica leaves. Six compounds were identified, including oleanolic acid (1), corosolic acid (2), trans-cinnamic acid (3), trans-sinapic acid (4), fisetin (5), and rutin (6). For the first time compound 5 was identified in this plant. The antibacterial effect of the two major compounds 1 and 6 was examined against Proteus mirabilis by applying the agar well diffusion method. Their minimum inhibitory concentrations (MICs) were then detected using broth microdilution assay. Rutin and oleanolic acid revealed MICs values ranging from 32 to 128 mu g/mL and 512 to 2048 mu g/mL, respectively. In addition, the antibiofilm action of the tested compounds was elucidated by crystal violet assay. Rutin exhibited antibiofilm activity, reducing the number of strong and moderate biofilm-forming isolates from 76.92% to 23.08%. Molecular analysis revealed that rutin downregulated mrpA, pmfA, and luxS gene expression in biofilm-forming P. mirabilis isolates. Further research is needed to explore the clinical implications of rutin.

Introduction: Multiple sclerosis (MS) as an autoimmune condition causes demyelination and neuronal loss.Materials and Methods: Forty-eight male adult dark agouti rats divided into six equal groups, group I functioned as control group. EAE was induced in rats of group II, III and IV; Flavonoids rich fraction (FRF) used as treatment of rats in group III; zinc oxide nanoparticles (ZnO-NPs) used as treatment of rats in group IV; Group V received Flavonoids rich fraction; Group VI received zinc oxide nanoparticles. The lumbar spinal cord was divided into three parts. The first part was for enzyme-linked immunosorbent assay (ELISA) and quantitative reverse transcription polymerase chain reaction (qRT-PCR) techniques. The second part of the spinal cord was for histopathological and immunohistochemical studies (H&E – MBP). The third part was for electron microscopic study.Results: This study demonstrates, for the first time, the beneficial effects of ZnO-NPs over FRF in ameliorating oxidative stress, modulating immune responses, reducing inflammation, and promoting neurodegenerative changes in a rat model of MS. Both FRF, and ZnO-NPs significantly suppressed the inflammatory activity detected in EAE model, through increasing IL-10 and decreasing lL-17 and TNF-α compared to the EAE group. H&E of sections of Group IV (ZnO-NPs treated group) revealed nearly normal architecture of lumbar section of spinal cord with myelinated nerve fibers and normal blood vessels. TEM examination of Flavonoids treated group (group III) showed less affected axons with little irregularity of myelin sheath. Group IV (ZnO-NPs treated group) TEM examination showed nearly normal nerve fibers axons with normal compact myelin sheath and normally arranged mitochondria with normal cristae.Conclusion: These outcomes might lessen symptoms, reduce the rate at which the illness progresses, enhance the quality of life for MS patients, and the potential intervention of ZnO-NPs as a therapeutic strategy for MS.

The global propagation of infections is a massive challenge in managing infected wounds. One of the most widely detected bacteria in wounds is Staphylococcus aureus. These bacteria possess multiple virulence factors, like biofilm formation, which hinder antibiotic treatment. Accordingly, it is vital to explore alternative therapeutics for managing these infections. We estimated the antibacterial and antibiofilm actions of N-acetylcysteine (NC). It revealed antibacterial action with minimum inhibitory concentration values of 256–2048 µg/mL. In addition, NC revealed antibiofilm action as exposed phenotypically from crystal violet assay. The NC diminished the percentages of strong and moderate biofilm-forming isolates from 75

The soil fungus Penicillium gladioli was identified by sequencing of the internal transcribed spacer (ITS) region. Gas chromatography-mass spectroscopy of the extract showed 50 compounds. Among the detected peaks, n-hexadecanoic acid showed the largest relative peak area (7.989% of the total ion chromatogram), followed by phenol, 2-methyl-5-(1-methylethyl) (6.543%). Regarding the in vivo anti-toxoplasma potential, there was an enhancement of the histological features of the liver of Swiss albino mice with a substantial decrease ( p < 0.05) in the inflammatory mediators, including cyclooxygenase-2, tumor necrosis factor-alpha, interleukin-6, and interleukin-1β. The colorimetric determination of the nitric oxide and malondialdehyde in the liver of the fungal extract-treated group revealed its antioxidant effect by significantly reducing ( p < 0.05) the oxidative stress markers. P. gladioli extract established antibacterial potential on P. aeruginosa bacteria with a minimum inhibitory concentration of 64 to 512 µg/ml. Moreover, it demonstrated antibiofilm potential using crystal violet assay and SEM. Also, 45% of the isolates displayed downregulation of the lasR , lecA , and pelA biofilm genes.

Cinacalcet hydrochloride (HCl), a calcium-sensing receptor agonist used to treat hyperparathyroidism, suffers from poor solubility, reducing its bioavailability. Recently, cinacalcet HCl has been probed for repurposing as antibacterial agent. This work investigates cinacalcet HCl's potential as an antibacterial agent and provides a formulation to improve the drug dissolution. Solid dispersion formulations using Poloxamer 407, with and without Soluplus®, were prepared via solvent evaporation and hot melt congealing methods. The resulting formulations were analyzed using differential scanning calorimetry, FTIR spectroscopy, X-ray powder diffraction, and dissolution studies. These formulations significantly enhanced cinacalcet HCl dissolution compared to the unprocessed form, achieving up to a 15-fold increase in Q5 (percent of cinacalcet HCl dissolved after 5 min). The dissolution efficiency rose from 28% for the pure drug to 94.8 and 87.8% for formulations F6 and F7, respectively. Microbiological evaluations confirmed the antibacterial effect of cinacalcet HCl, which was notably increased in the Poloxamer 407 and Soluplus® hybrid formulation (F7) with a MIC of 64-128 µg/ml. Antibiofilm activity was also observed, with qRT-PCR indicating downregulation of biofilm genes (icaA, icaD, and fnbA). This study introduces a cinacalcet HCl formulation prepared using a scalable, green approach, demonstrating significant potential for antimicrobial applications.

The spread of biofilm-forming multidrug-resistant pathogenic bacteria is an alarming public health issue requiring significant research. Drug repurposing is a novel approach to combating bacterial infections that is currently being studied. Here, we explored diclofenac sodium’s potential antibacterial and antibiofilm action on Staphylococcus epidermidis bacteria. Diclofenac sodium revealed antibacterial action on S. epidermidis isolates with minimum inhibitory concentrations of 500 to 2000 µg/mL. It also exposed antibiofilm action using the crystal violet assay and scanning electron microscope. Using qRT-PCR, diclofenac sodium has downregulated the expression of the biofilm genes (cna, fnbA, and ica) in 20% of the isolates. An animal model revealed the effect of diclofenac sodium on a systemic infection with S. epidermidis in mice. Diclofenac sodium has improved the liver, spleen, and kidney architecture. Molecular docking was used to explore the possible mechanism for the activity of diclofenac sodium against S. epidermidis, which revealed the high affinity of diclofenac sodium toward S. epidermidis protein and TcaR enzymes. Thus, diclofenac sodium could be a clinical solution for disseminating resistance among S. epidermidis and could be investigated for its combination with different antibiotics in future studies.

Abstract Carbapenem-resistant Klebsiella pneumoniae poses a severe risk to global public health, necessitating the immediate development of novel therapeutic strategies. The current study aimed to investigate the effectiveness of the green algae Arthrospira maxima (commercially known as Spirulina) both in vitro and in vivo against carbapenem-resistant K. pneumoniae. In this study, thirty carbapenem-resistant K. pneumoniae isolates were collected, identified, and then screened for their susceptibility to several antibiotics and carbapenemase production genes using PCR. Both bla KPC and bla OXA-48 genes were the most predominant detected carbapenemase genes in the tested isolates. The phytochemical profiling of A. maxima algal extract was conducted using LC–MS/MS in a positive mode technique. The minimum inhibitory concentrations (MIC) of the algal extract ranged from 500 to 1000 µg/mL. The algal extract also resulted in decreasing the membrane integrity and distortion in the bacterial cells as revealed by scanning electron microscope. The bioactive compounds that were responsible for the antibacterial action were fatty acids, including PUFAs, polysaccharides, glycosides, peptides, flavonoids, phycocyanin, minerals, essential amino acids, and vitamins. Moreover, A. maxima algal extract revealed an antibiofilm activity by crystal violet assay and qRT-PCR. A murine pneumonia model was employed for the in vivo assessment of the antibacterial action of the algal extract. A. maxima showed a promising antibacterial action which was comparable to the action of colistin (standard drug). This was manifested by improving the pulmonary architecture, decreasing the inflammatory cell infiltration, and fibrosis after staining with hematoxylin and eosin and Masson’s trichrome stain. Using immunohistochemical investigations, the percentage of the immunoreactive cells significantly decreased after using monoclonal antibodies of the tumor necrosis factor-alpha and interleukin six. So, A. maxima may be considered a new candidate for the development of new antibacterial medications. Graphical Abstract

In light of the propagation of resistance amongst bacteria, many infections are currently difficult to treat, with extremely high death and morbidity rates. Thus, we investigated the effect of using a combination of amoxicillin and broccoli ethanolic extract against Staphylococcus aureus infections. Broccoli or Brassica oleracea active metabolites' were to be identified and quantified using high-performance liquid chromatography (HPLC). The major detected compounds were chlorogenic acid and gallic acid. In vitro, the combination of amoxicillin with broccoli revealed fractional inhibitory concentration index of the amoxicillin-broccoli combination was less than 0.5 in 50% of the isolates, which indicates a synergetic potential. The in vivo study revealed a notable lessening in the bacterial count present in the examined tissues in the employed systemic infection model. The studied tissues were kidney, liver, and spleen. Moreover, the histological characteristics were found to have a remarkable enhancement in the combination-treated group. Thus, future clinical reports have to be employed to explicate the usefulness of such a combination in the clinical practice.