Hala I. Al-Daghistani

Pseudomonas aeruginosa is an opportunistic pathogen characterized by antimicrobial resistance and biofilm formation. This study investigated associations among biofilm intensity, antimicrobial resistance, pigment production, and selected virulence-associated genes in 91 clinical isolates collected in Jordan. Antimicrobial susceptibility was evaluated using disk diffusion, biofilm formation using the crystal violet microtiter assay, and selected virulence-associated genes using PCR. Biofilm formation was detected in 89 isolates (97.8%), with 41 (45.1%) and 31 (34.1%) exhibiting moderate and strong biofilm formation, respectively. Eleven isolates (12.1%) were multidrug-resistant, 14 (15.4%) extensively drug-resistant, and two (2.2%) classified pandrug-resistant based on the tested panel. Biofilm intensity was associated with resistance classification (p = 0.009) and correlated with resistance severity (Spearman’s ρ = 0.285, p = 0.006), the number of resistant antibiotics (ρ = 0.343, p = 0.0009), and the number of non-susceptible antibiotics (ρ = 0.379, p = 0.0002). In multivariable analysis, each increase in biofilm category was associated with higher odds of MDR/XDR/PDR classification (adjusted odds ratio = 2.50, 95% confidence interval: 1.24–5.02). Overall, greater biofilm formation intensity was associated with a higher antimicrobial resistance burden in this collection of clinical P. aeruginosa isolates, although further studies are needed to clarify its clinical significance.

Background: Helicobacter pylori colonizes the human gastric mucosa through outer membrane proteins (OMPs), particularly the adhesins BabA and BabB, which recognize host blood group antigens and mediate bacterial attachment and persistent colonization; importantly, structural and genetic studies have confirmed substantial polymorphisms in these adhesins that may affect their functional properties and contribute to differences in the clinical outcomes of infection. Therefore, the present study aimed to investigate the prevalence and genetic diversity of babA/babB variants in H. pylori isolates obtained from Jordanian patients and to evaluate their association with different gastric pathological conditions. Methods: A total of 106 gastric mucosal biopsies were collected from patients with gastric symptoms at two major hospitals in Jordan. Specimens underwent endoscopic evaluation, rapid urease testing (RUT), and histopathological examination according to the Sydney classification system. H. pylori detection was performed using real-time PCR targeting 16S rRNA and universal bab genes using novel primers designed to amplify both babA and babB variants. Conventional PCR was used for specific BabA and BabB gene amplification, followed by sequencing analysis of BabB variants. Results: A total of 106 patients were enrolled, with a mean age of 40.2 ± 15.9 years. H. pylori was detected in 76 (71.7%), 83 (78.3%), and 79 (74.5%) cases by the Rapid Urease Test (RUT), histopathology, and 16S rRNA qPCR, respectively. Using histopathology as the reference method, 16S rRNA qPCR demonstrated a sensitivity of 95.2%, compared with 91.6% for RUT. Among the 83 histopathology-positive cases, the universal bab gene was detected in 72 (86.7%), of which 53/72 (73.6%) were positive according to babB-specific PCR. Among patients with chronic gastritis, universal bab positivity increased progressively from 82.5% in mild to 88.5% in moderate and 93.3% in severe gastritis, whereas babB positivity increased from 50.0% to 76.9% and 80.0%, respectively. Sequencing of 40 babB-positive samples identified 27 distinct babB sequence profiles, with CHI-023 and LIM-008 being the most frequently identified reference sequence matches (10.0% each), highlighting substantial genetic diversity within the analyzed babB region. Conclusions: The findings demonstrate high diagnostic performance of 16S rRNA qPCR and RUT relative to histopathology, together with substantial genetic diversity within the analyzed babB region.

Helicobacter pylori is a well-established causative agent of gastritis, peptic ulcers, gastric adenocarcinoma, and primary gastric lymphoma. It colonizes the human stomach and expresses numerous virulent factors that influence disease progression. Among these factors is the cytotoxin vacA gene, which encodes the vacuolating capacity of the cytotoxin and plays a key role in the bacterium’s pathogenic potential. This study investigated the allelic diversity of the vacA among H. pylori strains infecting patients in Jordan with various gastric conditions and examined potential associations between vacA s-and m- genotypes, histopathological and endoscopic findings, and the development of gastric diseases. Gastric biopsies were collected from 106 patients at two hospitals in Jordan who underwent endoscopic examination. The collected biopsies for each patient were subjected to histopathological assessment, urease detection using the Rapid Urease Test (RUT), a diagnostic test for H. pylori, and molecular detection of the vacA gene and its s and m alleles. The histopathology reports indicated that 83 of 106 patients exhibited gastric disorders, of which 81 samples showed features associated with H. pylori infection. The RUT was positive in 76 of 106 with an accuracy of 93.8%. Real-time polymerase chain reaction (RT-PCR) targeting the 16S rRNA gene confirmed the presence of H. pylori in 79 of 81 histologically diagnosed cases as infected (97.5%), while the vacA gene was detected only in 75 samples (~95%). To explore genetic diversity, PCR-amplified fragments underwent sequence analysis of the vacA gene. The m-allele was detected in 58 samples (73%), the s-allele was detected in 45 (57%), while both alleles were not detected in 13% of samples. The predominant genotype combination among Jordanians was vacA s2/m2 (50%), significantly linked to mild chronic gastritis, followed by s1/m2 (35%) and s1/m1 (11.8%) which are linked to severe gastric conditions including malignancies. Age-and gender-related differences in vacA genotype were observed with less virulent s2m2 and s1m2 genotypes predominating in younger adults specially males, while the more virulent m1 genotypes were found exclusively in females and middle-aged patients. Genomic sequencing revealed extensive diversity within H. pylori, likely reflecting its long-standing co-evolution with human hosts in Jordan. This genetic variability plays a key role in modulating virulence and influencing clinical outcomes. Comprehensive characterization of vacA genotypic variations through whole-genome sequencing is essential to enhance diagnostic precision, strengthen epidemiological surveillance, and inform targeted therapeutic strategies. While this study highlights the significance of the vacA m and s alleles, future research is recommended in order to investigate the other vacA allelic variations, such as the i, d, and c alleles, to achieve a more comprehensive understanding of H. pylori pathogenicity and associated disease severity across different strains. These investigations will be crucial for improving diagnostic accuracy and guiding the development of targeted therapeutic strategies.

Background: The accurate and rapid diagnosis of infections is critical for effective and timely treatment. Misdiagnosis often leads to the prescription of antibiotics not targeting the causing agent of infection and thus be the possible development of multidrug resistance. This collectively worsens the condition and might lead to unnecessary intervention or death. The abundance of Pseudomonas spp. in healthcare-settings and the environment may lead to the inaccurate diagnosis of P. aeruginosa, making the treatment of its infections challenging. P. aeruginosa is a Gram-negative, opportunistic pathogen commonly linked to healthcare-associated infections. Its pathogenicity is attributed to several virulence factors correlated to enhanced survivability and colonization, invasion of the host tissues, and the development of multidrug resistance. When advanced diagnostic facilities are limited or unaffordable, the prescription of antibiotics solely relies on identifying the bacteria by culture-based methods. Objectives: This study aims to validate the accuracy of diagnosis of fifty clinical isolates preidentified as P. aeruginosa in three healthcare facilities in Jordan. Methods: The isolates were from infected areas of patients, including skin, wounds, ears, urine, and peritoneal cavities. Morphological and biochemical tests were performed, and the validation relied on the polymerase chain reaction (PCR) amplification of the 16S ribosomal ribonucleic acid (rRNA) gene. This molecular method is affordable for medical facilities with limited finances in contrast to advanced high-cost techniques. Results: The PCR confirmed that only 60% of the isolates were P. aeruginosa. All the confirmed isolates could produce different pigments and form biofilms. Conclusions: The high percentage of isolates mistakenly identified as P. aeruginosa raises concern about the suitability of prescribed antibiotics. The present study strongly recommends using advanced molecular methods to identify the pathogens. If conventional methods remain the only diagnostic option, this study recommends frequent external validation for tests in addition to performing an antibiotic susceptibility test to pinpoint the effective antibiotics against biofilm-producing P. aeruginosa.

Quorum sensing (QS) is a molecular communication mechanism among bacterial cells. It is critical in regulating virulence factors, motility, antibiotic resistance, and biofilm formation. Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen linked to healthcare-associated infections, food poisoning, and biofilm formation. Treating infections caused by pathogenic bacteria has become a challenge due to the development of multi-antibiotic resistance upon continuous exposure of bacteria to antibiotics. An alternative strategy to conventional antimicrobials to decrease the bacterial pathogenicity is QS inhibition, also known as quorum quenching. Using plant-derived compounds is an environmentally friendly strategy to block the bacterial QS and inhibit bacterial growth. Portulaca oleracea is a popular plant in different countries and is also used in traditional medicine. It is widely consumed raw in salads and as garnishes, though it can be cooked as a vegetarian dish. This study evaluates the antimicrobial activity of the methanolic extract of P. oleracea and its effectiveness in blocking or attenuating the QS of P. aeruginosa. The agar well diffusion method used for screening the antibacterial activity showed a significant growth inhibition of P. aeruginosa by the extract at 500 mg/mL with a minimum inhibitory concentration of 31.25 mg/mL. A bioindicator bacterium, Chromobacterium violaceum CV026, was used to determine the effect of the methanolic extract on the QS of P. aeruginosa. The results indicated a significant reduction in biofilm formation, pyocyanin production, and LasA staphylolytic activity. The phytochemical analysis by Gas Chromatography–Mass Spectrometry showed that the methanolic extract contained several phenols, alkaloids, esters, and other compounds previously reported to have antibacterial and antioxidant effects. These findings highlight the effectiveness of P. oleracea methanolic extract in attenuating the QS and virulence factors of P. aeruginosa. This study suggests that P. oleracea is an important source of natural antimicrobials and its use would be beneficial in food and pharmaceutical applications.

The interest in natural remedies in modern medicine has revealed potent disease-fighting agents, such asCUR(CUR) and nicotinamide (NIC), known for their antioxidant, anti-inflammatory, and immune-boosting properties. This study explores the antibacterial and antimelanoma potential of a CUR-NIC combination and its liposomal formulation. The antibacterial efficacy was assessed against skin infection-causing bacteria, Staphylococcus aureus, while cytotoxicity and migration assays were conducted on the melanoma B16 cancer cell line. The CUR-NIC combination (1:1) demonstrated an inhibition zone of 18±0.8 mm against S. aureus and a MIC of 31.25 µg/ml, outperforming CUR alone. Whereas Lip-CUR-NIC showed an inhibition zone of 12±0.8 mm against S. aureus and a MIC of 37.5 µg/ml. Notably, the cytotoxicity assay revealed that the CUR-NIC duo synergistically inhibited melanoma cell proliferation (CI<1). Liposomal preparations further enhanced this effect, with Lip-CUR-NIC showing a remarkably low IC50 of 8.5 ± 0.3 µM compared to CUR (IC50= 9.8 ± 2.2 µM) and NIC (IC50=135.95 ± 10.2 µM). These findings highlight the synergistic potential of CUR and NIC, especially in their liposomal form, offering a promising strategy for more effective cancer treatment.

The higher prevalence of cancer and the unmet need for antioxidant/anti‐inflammatory chemotherapeutic compounds with little side effect are of utmost importance. In addition, the increased likelihood of failure in clinical trials along with increasing development costs may have diminished the range of choices among newer drugs for clinical use. This has dictated the necessity to seek out novel medications by repurposing as it needs less time, effort, and resources to explore new uses of a current or unsuccessful medication. In this study, we examined the biological activity of 10 potential quinoline derivatives. Given the half‐maximal inhibitory concentration (IC₅₀ value) in lipopolysaccharide (LPS) induced inflammation of RAW264.7 mouse macrophages, all commercial FQs and selected quinolines (quinoline‐4‐carboxlic and quinoline‐3‐carboxylic acids) exerted impressively appreciable anti‐inflammation affinities versus classical NSAID indomethacin without related cytotoxicities in inflamed macrophages. Conversely, all 14 tested compounds lacked antioxidative DPPH radical scavenging capacities as compared to ascorbic acid. Gemifloxacin, considerably unlike markets FQs, indomethacin and quinoline derivatives, exerted exceptional and differential antiproliferation propensities in colorectum SW480, HCT116, and CACO2, pancreatic PANC1, prostate PC3, mammary T47D, lung A375, and melanoma A549 adherent monolayers using the sulforhodamine B colorimetric method versus antineoplastic cisplatin. All quinoline derivatives and gemifloxacin alike, but not levofloxacin, ciprofloxacin, or indomethacin, displayed substantially selective viability reduction affinities in prolonged tumor incubations of cervical HELA and mammary MCF7 cells. Specifically kynurenic acid (hydrate), quinoline‐2‐carboxylic acid, quinoline‐4‐carboxylic acid, quinoline‐3‐carboxylic acid, and 1,2‐dihydro‐2‐oxo‐4‐quinoline carboxylic acids possessed the most remarkable growth inhibition capacities against mammary MCF7 cell line, while quinoline‐2‐carboxylic acid was the only quinoline derivative with significant cytotoxicity on cervical HELA cancer cells. It is highly speculated that chelation with divalent metals via co‐planarity with close proximity of the COOH and the N atom could have the potential molecular mechanism for optimally promising repurposed pharmacologies. Conclusively, this study revealed the considerably profound repurposed duality of cytotoxicity and anti‐inflammation pharmacologies of quinoline derivatives. Activity‐guided structural modifications of the present nuclear scaffolds can be inherently linked to the betterment and enhancement of their repurposed pharmacologies.

The Dead Sea is unique compared to other extreme halophilic habitats. Its salinity exceeds 34%, and it is getting saltier. The Dead Sea environment is characterized by a dominance of divalent cations, with magnesium chloride (MgCl₂) levels approaching the predicted 2.3 M upper limit for life, an acidic pH of 6.0, and high levels of absorbed ultraviolet radiation. Consequently, only organisms adapted to such a polyextreme environment can survive in the surface, sinkholes, sediments, muds, and underwater springs of the Dead Sea. Metagenomic sequence analysis and amino acid profiling indicated that the Dead Sea is predominantly composed of halophiles that have various adaptation mechanisms and produce metabolites that can be utilized for biotechnological purposes. A variety of products have been obtained from halophilic microorganisms isolated from the Dead Sea, such as antimicrobials, bioplastics, biofuels, extremozymes, retinal proteins, colored pigments, exopolysaccharides, and compatible solutes. These resources find applications in agriculture, food, biofuel production, industry, and bioremediation for the detoxification of wastewater and soil. Utilizing halophiles as a bioprocessing platform offers advantages such as reduced energy consumption, decreased freshwater demand, minimized capital investment, and continuous production.

This is a comparative study to evaluate the effectiveness of six pomegranate peel extracts (PPEs) as antibacterial and antiproliferative agents. The Six PPEs were prepared using four solvent systems and each filtrate was concentrated to a gummy material to be used in the evaluation. The well‐diffusion method was used to evaluate their antimicrobial activity against bacteria typically associated with food spoilage: Escherichia coli, Pseudomonas aeruginosa, Salmonella typhimurium, Listeria monocytogenes, Staphylococcus epidermidis, Staphylococcus aureus, and three Bacillus species. The 3‐(4,5‐dimethylthiazol‐2‐yl)‐5‐(3‐carboxymethoxyphenyl)‐2‐(4‐sulfophenyl)‐2H‐tetrazolium (MTT) was used to evaluate the cytotoxicity against colorectal carcinoma cells (HCT116), prostate adenocarcinoma (PC3), ovarian cancer cells (SKOV‐3), and fibroblasts (MRC‐5). The antioxidant evaluation was done using the 2,2‐diphenyl‐1‐picrylhydrazyl‐hydrate (DPPH) assay. The pH of the water‐containing extracts was acidic and almost the same over 6 weeks. The six PPEs inhibited the bacterial growth in a comparable level to standard antibiotics. The effectiveness of each extract was dependent on the bacterial strain, and the Listeria showed a remarkable inhibition when exposed to the aqueous extract prepared at room temperature (RT). The aqueous (RT) and methanol PPEs had a significant antioxidant scavenging capability and a remarkable cytotoxic activity against the PC3 with half maximal inhibitory concentration (IC₅₀) of 0.1 μg/mL. The boiled aqueous extract exhibited antiproliferative activity against HCT116 with an IC₅₀ of 21.45 μg/mL. The effect on SKOV‐3 and fibroblasts was insignificant. With the exception of butanol, the antioxidant screening shows an inverse correlation between the polarity of the extraction solvent and the IC50 exhibited by the PPEs. The variation in the effectiveness of PPEs is suggested to be due to variable soluble bioactive compounds that may interact differently with different cells, though water‐containing extracts are promising antibacterial agents. The findings clearly show that pomegranate peel possessed the potential to be an eco‐friendly novel source for natural compounds that can be implemented in the food industry as a natural antimicrobial and natural food additive to prevent foodborne illnesses.

Campylobacteriosis, a foodborne illness, is one of the world′s leading causes of gastrointestinal illness. This study investigates the link between human campylobacteriosis and the consumption of potentially contaminated food with Campylobacter jejuni. Three hundred sixty samples were collected from humans, chicken cloaca, raw chicken meat, unpasteurized milk, and vegetables. The chickens were obtained from licensed and non-licensed slaughterhouses, and only the necks and wings were studied. Samples were enriched under microaerobic conditions then cultured on the modified charcoal cefoperazone deoxycholate agar. Bacteria was identified by staining, biochemical testing, and molecular identification by the polymerase chain reaction for the virulence genes; hipO, asp, dnaJ, cadF, cdtA, cdtB, and cdtC. The genomic homogeneity of C. jejuni between human and chicken isolates was assessed by the serological Penner test and the pulse field gel electrophoresis (PFGE). Campylobacter was not detected in the vegetables and pasteurized milk, though, only twenty isolates from chickens and clinical samples were presumed to be Campylobacter based on their morphology. The biochemical tests confirmed that five isolates were C. coli, and fifteen isolates were C. jejuni including two isolates from humans, and the remaining were from chickens. The colonization of C. jejuni in chickens was significantly lower in necks (6.66%) obtained from licensed slaughterhouses compared to those obtained from non-licensed slaughterhouses (33.3%). The antimicrobial susceptibility test showed that all identified C. jejuni isolates were resistant to antibiotics, and the majority of isolates (53.5%) showed resistance against six antibiotics, though, all isolates were resistant to ciprofloxacin, tetracycline, and aztreonam. The Penner test showed P:21 as the dominant serotype in isolates from humans, necks, and cloaca. The serohomology of C. jejuni from human isolates and chicken necks, wings, and cloaca was 71%, 36%, 78%, respectively. The PFGE analysis of the pattern for DNA fragmentation by the restriction enzyme Smal showed a complete genotypic homology of C. jejuni human isolates and chicken necks compared to partial homology with cloacal isolates. The study brings attention to the need for effective interventions to ensure best practices for safe poultry production for commercial food chain supply to limit infection with foodborne pathogens, including Campylobacter.