Prostatitis is an inflammatory condition that is related to multiple infectious agents, including bacteria and fungi. Traditional herbal extracts proved efficacious in controlling clinical symtpoms associated with prostatitis. In this context, the aim of the present study was to explore the efficacy of water extracts from S. virga-aurea, O. spinosa, P. boldus, E. angustifolium and P. niruri against bacterial (Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus cereus) and fungi strains (Candida albicans; C. tropicalis), involved in prostatitis. Additionally, anti-mycotic activities were tested against multiple species of dermatophytes (Tricholosporum rubrum, T. tonsurans, T. erinacei, Aspergillus crocatum, A. quadrifidum, A. gypseum, A. currey, A. insingulare), as well. Antioxidant effects were also evaluated in isolated rat prostate challenged with lipopolysaccharide (LPS), whereas phytochemical analyses were conducted to identify and quantify selected phenolic compounds, in the extracts. Finally, a bioinformatic analysis was conducted to predict putative human and microbial enzymes targeted by extracts’ phytocompounds and underlying the observed bio-pharmacological effects. The phytochemical analysis highlighted that rutin level could be crucial for explaining the highest antibacterial activity of P. boldus extract, especially against E. coli and B. cereus. On the other hand, in the E. angustifolium extract, catechin concentration could partially explain the highest efficacy of this extract in reducing lipid peroxidation, in isolated rat prostate stimulated with LPS. Concluding, the results of the present study showed antimicrobial and protective effects induced by water extracts of S. virga-aurea, P. boldus, E. angustifolium, P. niruri and O. spinosa, that are related, at least partially to phenolic composition of the phytocomplex. Additionally, the observed differences in extract efficacy supports the association of the aforementioned extracts, in order to improve the pharmacological spectrum. This could be in crucial for counteracting the burden of oxidative stress and inflammation occurring in bacterial prostatitis.

Industrial hemp (Cannabis sativa) is traditionally cultivated as a valuable source of fibers and nutrients. Multiple studies also demonstrated antimicrobial, anti-proliferative, phytotoxic and insecticide effects of the essential oil from hemp female inflorescences. On the other side, only a few studies explored the potential pharmacological application of polar extracts from inflorescences. In the present study, we investigated the water extract from inflorescences of industrial hemp Futura 75 variety, from phytochemical and pharmacological point of view. The water extract was assayed for phenolic compound content, radical scavenger/reducing, chelating and anti-tyrosinase effects. Through an ex vivo model of toxicity induced by lipopolysaccharide (LPS) on isolated rat colon and liver, we explored the extract effects on serotonin, dopamine and kynurenine pathways and the production of prostaglandin (PG)E₂. Anti-proliferative effects were also evaluated against human colon cancer HCT116 cell line. Additionally, antimycotic effects were investigated against Trichophyton rubrum, Trichophyton interdigitale, Microsporum gypseum. Finally, in silico studies, including bioinformatics, network pharmacology and docking approaches were conducted in order to predict the putative targets underlying the observed pharmacological and microbiological effects. Futura 75 water extract was able to blunt LPS-induced reduction of serotonin and increase of dopamine and kynurenine turnover, in rat colon. Additionally, the reduction of PGE₂ levels was observed in both colon and liver specimens, as well. The extract inhibited the HCT116 cell viability, the growth of T. rubrum and T. interdigitale and the activity of tyrosinase, in vitro, whereas in silico studies highlighting the inhibitions of cyclooxygenase-1 (induced by carvacrol), carbonic anhydrase IX (induced by chlorogenic acid and gallic acid) and lanosterol 14-α-demethylase (induced by rutin) further support the observed pharmacological and antimycotic effects. The present findings suggest female inflorescences from industrial hemp as high quality by-products, thus representing promising sources of nutraceuticals and cosmeceuticals against inflammatory and infectious diseases.

Introduction and Objectives: Klebsiella pneumoniae is a major pathogen implicated in nosocomial infections that is known to spread easily, and it is frequently associated with resistance to the highest-priority critically important antimicrobials. The purpose of this work was to determine the genetic diversity (MLST) among selected carbapenem-and ESBL-producing K. pneumoniae isolates from human urinary infections.

Materials and Methods: Forty-nine cefotaxime/ceftazidime-resistant K. pneumoniae isolates were obtained aleatory from urocultures from patients of a Portuguese hospital during June 2017-July 2018. Identification was performed by MALDI-TOF MS. Antimicrobial susceptibility against 13 antibiotics was analyzed by disk diffusion test. Detection of ESBLs and other resistance and integron genes, and molecular typing (for selected isolates) was performed by PCR/sequencing.

Results: ESBL-production was detected in 26.5% of the isolates (13/49), most of them associated with CTX-M-15 enzyme (n=10). It is important to note that all ESBL-positive and negative isolates carried the KPC_2/3 gene. Regarding the ESBL-producing Klebsiella pneumoniae isolates, different sequence types (ST) were identified (ST/phylogenetic-group/β-lactamase): ST15/CTX-M-15, SHV-28; ST15/CTX-M-15, SHV-12; ST280/CTX-M-15, SHV-27; ST280/SHV-27 and ST147/SHV-12. Additionally, the selected ESBL-negative isolates were typed as ST15/SHV-28, ST34/SHV-26 and ST348/SHV-11.

Conclusion: These findings indicate the genetic diversity among urinary infections isolates in our hospital. Furthermore, the KPC_2/3 is the main mechanism of carbapenem resistance in K. pneumoniae isolates, frequently associated with CTX-M-15 enzyme.

The main problem in the treatment of nosocomial infections is spreading of gram-negative carbapenem-resistant microorganisms. Therefore, it is important to monitor the sensitivity of nosocomial infections to antibiotics from the carbapenem group and to determine the mechanisms of the microorganisms resistance.

Methods: During the period from 2014 to 2020, 98 strains isolated from blood and cerebrospinal fluid of the children in intensive care units were selected. Antibiotic sensitivity was determined by broth microdilution. Strains of P. aeruginosa with a minimum inhibitory concentration of meropenem> 2 mg / L and / or imipenem> 0.001 mg / L, in K. pneumoniae and A. baumannii, meropenem> 2 mg / L and / or imipenem> 2 mg / L, other strains were classified as susceptible to carbapenems. Carbapenemases were detected by real-time multiplex polymerase chain reaction (PCR).

Results: Among the studied strains there were 62 (63%) K. pneumoniae strains, 22 (23%) P. aeruginosa strains and 14 (14%) A. baumannii strains. Among all isolates of K. pneumoniae, 28 (45%) were classified as resistant to carbapenems, the other 34 (55%) strains were classified as susceptible. Among the studied strains of P. aeruginosa, 16 (73%) were insensitive to carbapenems, and 6 (27%) isolates were susceptible. Of the A. baumannii strains studied, 10 (71%) were in the carbapenem-insensitive group and 4 (29%) were in the carbapenem-susceptible group.

In the process of studying the mechanisms of resistance to carbapenems, the following results were obtained. In K. pneumoniae isolates, the cause of resistance to carbapenems was bla_OXA-48 carriage (n = 25, 89%). The presence of bla_NDM and bla_KPC was not identified. 11 (69%) P. aeruginosa isolates had bla_VIM. The A. baumannii strains had carbapenemase OXA-40 (n = 5, 50%) and OXA-23 (n = 3, 30%).

Conclusions: In most cases, resistance to carbapenem antibiotics in the studied strains isolated from blood and cerebrospinal fluid of children in intensive care units was due to the presence of various carbapenemases. For K. pneumoniae, OXA-48 was the only determinant of resistance. P. aeruginosa resistance was associated with VIM group carbapenemases. The A. baumannii strains contained carbapenemases OXA-40 and OXA-23.

Coronilla species, belonging to the Coronilla genus (Fabaceae), have long been used in traditional medicine for treating cold, diabetes, pain, and as cardiotonics. The goal of the present study was to explore the phytochemical composition and pharmaco-toxicological properties of C. minima. In this regard, phenolic content, scavenging/reducing properties and antimicrobial activity toward pathogen bacterial (Escherichia coli, Pseudomonas aeruginosa, Bacillus cereus, Staphylococcus aureus) and fungal strains (Candida albicans, C. tropicalis, Aspergillus tubigensis and A. minutus) were investigated. Extract effects on human colon cancer HCT116 cell viability were also assayed. Finally, a bioinformatics approach was conducted with the aim to identify putative microbial and human protein targets underlying antibacterial, antimycotic, and antiproliferative effects. Phytochemical investigation suggested that water extract is richer in terms of total flavonoid and phenol content, whereas the hydroalcoholic extract was revealed to be more potent as antioxidant agent. According to bioinformatics analysis, the antibacterial activity of the hydroalcoholic extract could be related to its content in resveratrol. The presence of resveratrol could also explain the hydroalcoholic extract efficacy in reducing HCT116 cell viability. In conclusion, the present study represents the first phytochemical and bio-pharmacological investigation about C. minima. Like other plants belonging to the Fabaceae family, C. minima revealed a good source of resveratrol, which could explain, albeit partially, the efficacy of the hydroalcoholic extract as antimicrobial, antioxidant, and antiproliferative agent.

Tricholosporum goniospermum (Bres.) Guzmán ex T.J. Baroni is an excellent edible mushroom whose compounds and biological properties are still unknown. In this study, n-hexane, ethyl acetate and methanol extracts from fruiting bodies and liquid-cultured mycelia were compared for the analysis of phenolic compounds, the evaluation of scavenger (DPPH, ABTS) and reducing (CUPRAC, FRAP) activities, and the enzyme inhibition of α-amylase, acetylcholinesterase (AChE), butyrylcholinesterase (BChE) and tyrosinase. Additionally, T. goniospermum extracts were evaluated for antibacterial and antimycotic activities against Gram+ and Gram− bacteria, and clinical yeast and fungal dermatophytes. Finally, based on the extract content in phenolic compounds, in silico studies, including the docking approach, were conducted to predict the putative targets (namely tyrosinase, lanosterol-14-α-demethylase, the multidrug efflux system transporters of E. coli (mdtK) and P. aeruginosa (pmpM), and S. aureus β-lactamase (ORF259)) underlying the observed bio-pharmacological and microbiological effects. The methanolic extract from mycelia was the richest in gallic acid, whereas the ethyl acetate extract from fruiting bodies was the sole extract to show levels of catechin. Specifically, docking runs demonstrated an affinity of catechin towards all docked proteins, in the micromolar range. These in silico data are consistent, at least in part, with the highest activity of ethyl acetate extract as an antimicrobial and anti-tyrosinase (554.30 mg KAE/g for fruiting bodies and 412.81 mg KAE/g for mycelia) agent. The ethyl acetate extracts were also noted as being the most active (2.97 mmol ACAE/g for fruiting bodies and 2.25 mmol ACAE/g for mycelia) on α-amylase. BChE inhibitory activities varied from 2.61 to 26.78 mg GALAE/g, while the tested extracts were not active on AChE. In conclusion, all mushroom extracts tested in this study had potent antimicrobial activities. Particularly, among the tested extracts, the ethyl acetate extract showed the highest efficacy as both an antimicrobial and anti-tyrosinase agent. This could be related, albeit partially, to its content of catechin. In this regard, the bioinformatics analyses showed interactions of catechin with tyrosinase and specific microbial proteins involved in the resistance to chemotherapeutic drugs, thus suggesting innovative pharmacological applications of T. goniospermum extracts.

In the present study, we investigated the water extract of Harpagophytum procumbens DC. ex Meisn. in an experimental model of inflammatory bowel diseases (IBDs). Additionally, a microbiological investigation was carried out to discriminate the efficacy against bacterial and fungal strains involved in IBDs. Finally, an untargeted proteomic analysis was conducted on more than one hundred colon proteins involved in tissue morphology and metabolism. The extract was effective in blunting the production of oxidative stress and inflammation, including serotonin, prostaglandins, cytokines, and transcription factors. Additionally, the extract inhibited the growth of Candida albicans and C. tropicalis. The extract was also able to exert a pro-homeostatic effect on the levels of a wide plethora of colon proteins, thus corroborating a protective effect. Conversely, the supraphysiological downregulation of cytoskeletal-related proteins involved in tissue morphology and antimicrobial barrier function suggests a warning in the use of food supplements containing H. procumbens extracts.

Bacteria regulate their virulence factor production and biofilm formation through an intercellular communication system mediated by the binding of signaling molecules to QS receptors such as LasR. A range of natural and synthetic brominated furanones known as fimbrolides and their dihydropyrolones counterparts have been found to act as inhibitors of QS-dependent bacterial phenotypes. In this study, a range of dihydropyrrolone (DHP) analogues were synthesized via the lactone-lactam conversion of lactone intermediates followed by the formation of novel acetylene analogues of dihydropyrrolones. Fifteen novel alkyne analogues of DHPs with various substitution patterns and aliphatic chain lengths were successfully synthesised via lactamisation and Sonogashira coupling reactions in moderate to high yields. The Sonogashira reaction was carried out with DHPs and alkynes in the presence of CuI, palladium catalyst PdCl₂(PPh₃)₂ and TEA. Biological testing demonstrated that several compounds showed low to moderate activity against the P. aeruginosa MH602 reporter strain with little bactericidal effect. The present study represents the first application of the Sonogashira reaction to DHP scaffolds for the synthesis of novel bacterial QS inhibitors.

Bacteriocin can be defined as ribosomal synthesized antimicrobial polypeptides or proteins secreted by bacteria chiefly by lactic acid bacteria. Natural food preservation is the method in which we use naturally produced antimicrobial compounds or normal-flora as a food preservative to prevent food spoilage microorganisms and to extend food shelf life. Currently, some ambitious rational motives like augmentative of consumers demand to get a new and fresh like foods, potential health hazards of artificial food preservative and accumulation of irrepressible food additive have to lead to the development of an alternative food preservation technology to maintain the freshness of food products. Therefore, bacteriocins are emerging as the very likely natural alternative preservative to chemical preservative and widely accepted as safe food preservative though out worldwide. Due to consumers’ demand and awareness on health effect and benefits of fermented foods and strict government legislation on foods , applications of bacteriocins novel natural food preservative initiates searching for the design of novel technologies used in the food industry. Therefore, the main objective of review paper is to explore the general background and application of bacteriocin as a novel and promising food preservative.

Introduction: Resistance to B-lactam antibiotics has become increasingly prevalent since the introduction of antibiotics. Each year in the United States approximately two million people acquire bacterial infections that are resistant to antibiotics. A common mechanism for B-lactam resistance is the production of B-lactamases that hydrolyze the B-lactam ring, thus rendering the drugs inactive. Today there are more than 2000 B-lactamases, but this study will focus on the B1 subclass known as metallo-B-lactamases (MBLs). MBLs are capable of inactivating all B-lactam antibiotics, except monobactams, and do not have any known clinical inhibitors. Thus, the development of MBL inhibitors is crucial. Most studies focus on the three most clinically relevant MBLs which include: Imipenemase MBL (IMP-1); Verona integrin-encoded MBL (VIM-2); and New Delhi MBL (NDM-1). Over time each of these MBLs have evolved and now have many variants whose inhibition is poorly studied. The variant IMP-78 will be the focus of this study due to its two mutations (S262G/V67F) near the active site. The primary purpose of this study is to investigate whether an inhibitor of IMP-1 is capable of having the same mechanism of inhibition with the variant IMP-78. Results: Six previously published novel MBL inhibitors were chosen for this study. The native state electrospray ionization mass spectrometry (ESI-MS) showed that 3 out of 6 inhibitors were capable of binding to both IMP-1 and IMP-78. While the remaining 3 inhibitors were capable of binding only to IMP-1 and not to IMP-78. The preliminary molecular modeling results suggest a difference in the size of the binding pockets of IMP-1 and IMP-78. Future work: As this study is currently ongoing, there is future work to be completed. To investigate whether one or both amino acid mutations in IMP-78 prevent inhibitor binding, IMP-6 (S262G) and IMP-10 (V67F) will be analyzed using native state ESI-MS. The molecular modeling and molecular dynamic (MD) simulations will be completed on each inhibitor with IMP-1 and IMP-78. Molecular modeling and MD simulations will be performed for the 3 inhibitors that bind IMP-1 but not IMP-78 using IMP-6 and IMP-10. The binding affinity of each inhibitor for IMP-1 and IMP-78 will be determined using ITC. Lastly, microbiological studies (MIC’s) will be conducted.