Pomegranate peel: Bioactivities as antimicrobial and cytotoxic agents.

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.

8-Amino-7-(aryl/hetaryl)fluoroquinolones. An emerging set of synthetic antibacterial agents.

This study reports the synthesis of seven new 8-amino-7-(aryl/hetaryl)fluoroquinolones and their antibacterial activity against 10 bacteria associated with microbial infections and foodborne illnesses. These fluoroquinolones are prepared via the reactions of selected aryl(hetaryl)boronic acids with ethyl-7chloro-6-fluoro-8-nitroquinolone-3-carboxylate, under Suzuki-Miyaura cross-coupling conditions. Nitro group reduction of the latter resulted in the corresponding 8-aminoquinolone-3-esters which upon hydrolysis formed the respective 8-amino-7-(aryl/hetaryl)-quinolone-3-carboxylic acids. The latter compounds were tested against selected Gram-negative bacteria (Escherichia coli, Salmonella typhimurium, Pseudomonas aeruginosa, Acinetobacter baumannii, and Klebsiella pneumonia) and Gram-positive bacteria (Enterococcus feacalis, Listeria monocytogenes, Streptococcus agalactiae, Staphylococcus epidermidis, and Staphylococcus aureus). The tested fluoroquinolones showed a significant antimicrobial activity against most of the tested bacterial strains. The antimicrobial activity of some of the tested compounds were comparable to or higher than a wide range of standard antibiotics including ampicillin, ciprofloxacin, and imipenem. The results highlight the new synthesized 8-amino-7-(aryl/hetaryl)fluroquinolones as promising candidates for new antimicrobial drugs to treat bacterial infections. This study highlights that the newly synthetic 8-amino-7-(aryl/hetaryl)fluroquinolones are promising candidates for new antimicrobial drugs to treat human diseases including foodborne illnesses.

Characterizing Bacillus globigii as a Bacillus anthracis surrogate for wastewater treatment studies and bioaerosol emissions.

This study characterized Bacillus globigii (BG) as a Bacillus anthracis Sterne (BAS) surrogate for wastewater treatment-related studies of UV inactivation, adsorption onto powdered activated carbon (PAC), and bioaerosol emission. The inactivation of BG was faster than that of BAS in DI water (pseudo first-order rate constants of 0.065 and 0.016 min-1 respectively) and in PBS solution (0.030 and 0.005 min-1 respectively). BG was also removed more quickly than BAS by PAC adsorption in DI (0.07 and 0.05 min-1 respectively) and in PBS (0.09 and 0.04 min-1 respectively). In DI, BG aggregated more (P < 0.05) than BAS when the pH was 7 or greater but there were no statistically significant differences in NaCl solution. Spore aggregation was also studied with extended Derjaguin-Landau-Verwey-Overbeek (XDLVO) models. Less than 1% of all spores were released as bioaerosols, and there was no significant difference (P > 0.05) in emission between BG and BAS. To the author's knowledge, this study is the first to demonstrate that BG is a suitable surrogate for BAS for bioaerosol emissions, but a poor surrogate for both UV inactivation and PAC adsorption. These results can be used to understand the ability of BAS to act as a surrogate for BA Ames because of its genetic and morphological similarities with BAS.

Genetic Signature and Serocompatibility Evidence for Drug Resistant Campylobacter jejuni.

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.

Protein phosphatase 2A and Cdc7 kinase regulate the DNA unwinding element-binding protein in replication initiation.

The DNA unwinding element (DUE)-binding protein (DUE-B) binds to replication origins coordinately with the minichromosome maintenance (MCM) helicase and the helicase activator Cdc45 in vivo, and loads Cdc45 onto chromatin in Xenopus egg extracts. Human DUE-B also retains the aminoacyl-tRNA proofreading function of its shorter orthologs in lower organisms. Here we report that phosphorylation of the DUE-B unstructured C-terminal domain unique to higher organisms regulates DUE-B intermolecular binding. Gel filtration analyses show that unphosphorylated DUE-B forms multiple high molecular weight (HMW) complexes. Several aminoacyl-tRNA synthetases and Mcm2-7 proteins were identified by mass spectrometry of the HMW complexes. Aminoacyl-tRNA synthetase binding is RNase A sensitive, whereas interaction with Mcm2-7 is nuclease resistant. Unphosphorylated DUE-B HMW complex formation is decreased by PP2A inhibition or direct DUE-B phosphorylation, and increased by inhibition of Cdc7. These results indicate that the state of DUE-B phosphorylation is maintained by the equilibrium between Cdc7-dependent phosphorylation and PP2A-dependent dephosphorylation, each previously shown to regulate replication initiation. Alanine mutation of the DUE-B C-terminal phosphorylation target sites increases MCM binding but blocks Cdc45 loading in vivo and inhibits cell division. In egg extracts alanine mutation of the DUE-B C-terminal phosphorylation sites blocks Cdc45 loading and inhibits DNA replication. The effects of DUE-B C-terminal phosphorylation reveal a novel S phase kinase regulatory mechanism for Cdc45 loading and MCM helicase activation.