Translating phage therapy into the clinic: Recent accomplishments but continuing challenges.

Phage therapy is a medical form of biological control of bacterial infections, one that uses naturally occurring viruses, called bacteriophages or phages, as antibacterial agents. Pioneered over 100 years ago, phage therapy nonetheless is currently experiencing a resurgence in interest, with growing numbers of clinical case studies being published. This renewed enthusiasm is due in large part to phage therapy holding promise for providing safe and effective cures for bacterial infections that traditional antibiotics acting alone have been unable to clear. This Essay introduces basic phage biology, provides an outline of the long history of phage therapy, highlights some advantages of using phages as antibacterial agents, and provides an overview of recent phage therapy clinical successes. Although phage therapy has clear clinical potential, it faces biological, regulatory, and economic challenges to its further implementation and more mainstream acceptance.

How Nano-ZnO Affect Tomato Fruits (Solanum lycopersicum L.)? Analysis of Selected Fruit Parameters.

Tomato (Solanum lycopersicum L.), as one of the most valuable horticulture crops, was chosen to investigate the effect of nanoparticles (NPs) in the form of nano-ZnO combined with conventional fertilizer on the quality of tomato fruits, including their antioxidant potential (total antioxidant activity, lycopene and ß-carotene content), sugars content and allergenic potential (profilin and Bet v 1 content). Nano-ZnO was implemented during plant cultivation, applied by foliar spraying or directly via soil, at three different concentrations (50, 150 and 250 mg/L). The obtained results suggest that the usage of NPs during tomato plant cultivation had minor impacts on parameters such as total antioxidant activity or the content of selected allergens. Even though the total antioxidant activity was not affected by nano-ZnO, the malondialdehyde activity (MDA) content was notably decreased in fruits under nano-ZnO treatment. The content of lycopene and ß-carotene was significantly affected by the use of nano-ZnO. Moreover, the usage of nano-ZnO significantly increased the total sugar content in fruits treated with nanoparticles via foliar spraying. Based on the obtained results, it can be stated that nano-ZnO, regardless of the method of application, significantly affected tomato fruits which can be beneficial for fruit production.

The Antioxidant Potential of Tomato Plants (Solanum lycopersicum L.) under Nano-ZnO Treatment.

Tomato (Solanum lycopersicum L.) is one of the most valuable horticulture crops, consumed in both its raw and processed forms. To increase yield and efficiency, conventional and organic fertilizers are utilized in modern agriculture. Traditional fertilizers increase crop yield but are harmful to the environment. These circumstances motivate the pursuit of an alternate solution. The purpose of this research was to investigate how the application of nanoparticles (nano-ZnO) combined with conventional fertilizer influence tomato plants' development, including the antioxidant potential of cultivated plants. Three factors such as different types of cultivars, dosage of applied nano-ZnO solution and the method of nanoparticles application were implemented. Multiple analysis of selected antioxidants content and their activities such as malondialdehyde (MDA), flavonoids, polyphenols, ascorbic acid, peroxidase (POX), superoxide dismutase (SOD) or catalase (CAT) were analyzed. The obtained data exhibited that all examined parameters were strongly dependent on three implemented factors: concentration of nano-ZnO suspension, the type of cultivated tomato and the method of nanoparticles application. For instance, the accumulation of MDA in cultivated plants was different among plants under nanoparticles treatment, but in one specific case (Malinowy Bossman cultivar treated with 50 mg/L nano-ZnO suspension) the content of this marker was decreased by 34% in comparison to the corresponding control. Nevertheless, the results presented in this study showed that the usage of certain doses of nano-ZnO suspension may increase the antioxidant potential of tomato plants.

The Effect of Nano-ZnO on Seeds Germination Parameters of Different Tomatoes (Solanum lycopersicum L.) Cultivars.

The agriculture sector faces numerous problems. One of the beforementioned problems relates to the proper crop plants' fertilization. The conventional bulk fertilizers are becoming less effective and have a negative impact on the environment. Nanomaterials such as zinc oxide nanoparticles (ZnO NPs) are widely used in various sectors such as medicine or electronics. Several studies indicate that nano-ZnO may likewise be considered as a potential nanofertilizer. In present research, an attempt was made to study the influence of two different sized ZnO NPs (<50 nm and <100 nm) on the seed germination of chosen tomato (Solanum lycopersicum) cultivars. The seeds of three cherry tomato cultivars were placed on a Petri dish with the NPs suspensions (0, 50, 150, and 250 mg/L) in order to examine the influence on germination parameters at a certain size of NPs and at a chosen concentration. In addition, within this study, we verified that the implicated conditions have the exact impact on all three cultivars. The obtained results indicate that all the factors affect the seed sprouting, however, this process mainly depends on the type of tomato cultivar and the size of the used nanoparticles. The parameter of the germination percentage (GP) was the only of the assumed factors that did not influence it significantly. Nevertheless, the values of other examined parameters such as the MGT, GRI, CVG, or VI depend strongly on all assumed features including the type of chosen cultivar. The obtained results vary significantly between all cultivars which indicates that the plants from the same family may require different conditions for optimal growth. In this research the <50 nm ZnO nanoparticles had more beneficial influence on sprouting parameters then parallelly used <100 nm ZnO nanoparticles.

Phage Therapy: The Pharmacology of Antibacterial Viruses.

Pharmacology can be differentiated into two key aspects, pharmacodynamics and pharmacokinetics. Pharmacodynamics describes a drug's impact on the body while pharmacokinetics describes the body's impact on a drug. Another way of understanding these terms is that pharmacodynamics is a description of both the positive and negative consequences of drugs attaining certain concentrations in the body while pharmacokinetics is concerned with our ability to reach and then sustain those concentrations. Unlike the drugs for which these concepts were developed, including antibiotics, the bacteriophages (or 'phages') that we consider here are not chemotherapeutics but instead are the viruses of bacteria. Here we review the pharmacology of these viruses, particularly as they can be employed to combat bacterial infections (phage therapy). Overall, an improved pharmacological understanding of phage therapy should allow for more informed development of phages as antibacterial 'drugs', allow for more rational post hoc debugging of phage therapy experiments, and encourage improved design of phage therapy protocols. Contrasting with antibiotics, however, phages as viruses impact individual bacterial cells as single virions rather than as swarms of molecules, and while they are killing bacteria, bacteriophages also can amplify phage numbers, in situ. Explorations of phage therapy pharmacology consequently can often be informed as well by basic principles of the ecological interactions between phages and bacteria as by study of the pharmacology of drugs. Bacteriophages in phage therapy thus can display somewhat unique as well as more traditional pharmacological aspects.

Integrative omics analysis of Pseudomonas aeruginosa virus PA5oct highlights the molecular complexity of jumbo phages.

Pseudomonas virus vB_PaeM_PA5oct is proposed as a model jumbo bacteriophage to investigate phage-bacteria interactions and is a candidate for phage therapy applications. Combining hybrid sequencing, RNA-Seq and mass spectrometry allowed us to accurately annotate its 286,783 bp genome with 461 coding regions including four non-coding RNAs (ncRNAs) and 93 virion-associated proteins. PA5oct relies on the host RNA polymerase for the infection cycle and RNA-Seq revealed a gradual take-over of the total cell transcriptome from 21% in early infection to 93% in late infection. PA5oct is not organized into strictly contiguous regions of temporal transcription, but some genomic regions transcribed in early, middle and late phases of infection can be discriminated. Interestingly, we observe regions showing limited transcription activity throughout the infection cycle. We show that PA5oct upregulates specific bacterial operons during infection including operons pncA-pncB1-nadE involved in NAD biosynthesis, psl for exopolysaccharide biosynthesis and nap for periplasmic nitrate reductase production. We also observe a downregulation of T4P gene products suggesting mechanisms of superinfection exclusion. We used the proteome of PA5oct to position our isolate amongst other phages using a gene-sharing network. This integrative omics study illustrates the molecular diversity of jumbo viruses and raises new questions towards cellular regulation and phage-encoded hijacking mechanisms.

Physicochemical and Antifungal Properties of Clotrimazole in Combination with High-Molecular Weight Chitosan as a Multifunctional Excipient.

Chitosans represent a group of multifunctional drug excipients. Here, we aimed to estimate the impact of high-molecular weight chitosan on the physicochemical properties of clotrimazole-chitosan solid mixtures (CL-CH), prepared by grinding and kneading methods. We characterised these formulas by infrared spectroscopy, differential scanning calorimetry, and powder X-ray diffractometry, and performed in vitro clotrimazole dissolution tests. Additionally, we examined the antifungal activity of clotrimazole-chitosan mixtures against clinical Candida isolates under neutral and acid conditions. The synergistic effect of clotrimazole and chitosan S combinations was observed in tests carried out at pH 4 on Candida glabrata strains. The inhibition of C. glabrata growth reached at least 90%, regardless of the drug/excipient weight ratio, and even at half of the minimal inhibitory concentrations of clotrimazole. Our results demonstrate that clotrimazole and high-molecular weight chitosan could be an effective combination in a topical antifungal formulation, as chitosan acts synergistically with clotrimazole against non-albicans candida strains.

[1,3]/[1,4]-Sulfur atom migration in ß-hydroxyalkylphosphine sulfides.

ß-Hydroxyalkylphosphine sulfides undergo [1,3]- or [1,4]-sulfur atom phosphorus-to-carbon migration in the presence of Lewis or Brønsted acids. The direction of sulfur atom migration depends on the type of acid used for the reaction. In the presence of a Brønsted acid, mainly [1,3]-rearrangement is observed, whereas a Lewis acid catalyzes the [1,4]-sulfur migration. To gain insight into the mechanism of these transformations, the stereochemistry of these rearrangements have been tested, along with the conduction of some control experiments and DFT calculations.

Detection of Polish clinical Aspergillus fumigatus isolates resistant to triazoles.

We studied the presence of triazole resistance of 121 Aspergillus fumigatus clinical isolates collected in two Polish cities, Warsaw and Wroclaw, to determine if resistance is emerging in our country. We identified five itraconazole resistant isolates (4.13%) carrying the TR34/L98H alteration in Cyp51A gene, four of which were cross-resistant to posaconazole and one to voriconazole. One isolate was intermediate susceptible to itraconazole and harbored no Cyp51A alterations. The study confirms the presence of azole resistant A. fumigatus strains in Poland at a level that is comparative to other European countries.

Detection of dermatophytes in human nail and skin dust produced during podiatric treatments in people without typical clinical signs of mycoses.

Pedicures are the most common cosmetic foot treatment. Many pedicurists and podiatrists suffer from respiratory infections and diseases such as asthma, sinusitis, chronic cough and bronchitis. Skin and nail dust may play an important role in the development of occupational diseases and the transmission of mycosis to other clients. To examine the presence of dermatophytes in nail and skin dust produced during podiatric treatments of people without typical symptoms of mycosis and to assess the epidemiological hazards of tinea pedis for podiatrists as well as other clients. Seventy-seven samples underwent direct microscopy and culture. The results of direct microscopy were positive in 28/77 samples (36.36%) and doubtful in 3/77 (3.9%). Fungi were cultured from 36/77 samples (46.75%), including 8/77 (10.3%) positive for dermatophytes (Trichophyton rubrum-6 isolates and Trichophyton mentagrophytes-2). Material collected during podiatric treatments is potentially infected by pathogenic fungi; thus, there is a need to protect both workers who perform such treatments, as well as other clients, to prevent the transmission of pathogens in the Salon environment. Exposure to this occupational hazard may increase not only the risk of respiratory infections but also increase asthmatic or allergic reactions to Trichophyton.

New monomeric and dimeric uridinyl derivatives as inhibitors of chitin synthase.

This study described the synthesis and in vitro evaluation of eight new derivatives of uridine as antifungal agents and inhibitors of chitin synthase. Dimeric uridinyl derivatives synthesized by us did not exhibit significant activity. One of the studied monomeric derivative, 5'-(N-succinyl)-5'-amino-5'-deoxyuridine methyl ester (compound 7) showed activities against several fungal strains (MIC range 0.06-1.00 mg/mL) and inhibited chitin synthase from Saccharomyces cerevisiae (IC50=0.8mM). Moreover compound 7 exhibited synergistic interaction with caspofungin against Candida albicans (FIC index=0.28).

In vitro and in vivo antibacterial activity of environmental bacteriophages against Pseudomonas aeruginosa strains from cystic fibrosis patients.

The goal of the study was to determine the relationship between in vitro/in vivo efficacy of environmental Pseudomonas phages and certain phenotypical properties of Pseudomonas aeruginosa (PA) strains. We studied the diversity between particular isolates and determined phage sensitivity in vitro and in vivo in the Galleria mellonella insect model. Twenty-eight lytic bacteriophages specific for PA were tested against 121 CF PA isolates including 29 mucoid PA strains. Most strains from cystic fibrosis (CF) patients were lysed by at least three phages (93.6 %), but completely insensitive strains were also present (6.4 %). Two phages PA5oct and KT28 exhibited high rates of lytic potency on 55-68 % of PA strains (72-86 % of mucoid isolates). We further explored phage activity against six PA strains (CF and non-CF) in vitro, comparing clonal differences in phage susceptibility with bacterial properties such as the ability to form biofilms, mucosity, twitching motility, and biochemical profiles. We observed the relationship between variation in phage susceptibility and Fourier transform infrared spectroscopy (FTIR) analysis in the spectra window of carbohydrates. The protective efficacy of two selected phages against PA PAO1 and 0038 infection was confirmed in vivo in G. mellonella larvae. Generally, the wax moth model results confirmed the data from in vitro assays, but in massive infection of CF isolates, the application of lytic phages probably led to the release of toxic compound causing an increase in larvae mortality. We assumed that apart of in vitro phage activity testing, a simple and convenient wax moth larvae model should be applied for the evaluation of in vivo effectiveness of particular phage preparations.

The Effect of High-Temperature Heating on Amounts of Bioactive Compounds and Antiradical Properties of Refined Rapeseed Oil Blended with Rapeseed, Coriander and Apricot Cold-Pressed Oils.

Cold-pressed oils are rich sources of bioactive substances, which may protect triacylglycerols from degradation during frying. Nevertheless, these substances may decompose under high temperature. This work considers the content of bioactive substances in blends and their changes during high-temperature heating. Blends of refined rapeseed oil with 5% or 25% in one of three cold-pressed oils (rapeseed, coriander and apricot) were heated at 170 or 200 °C in a thin layer on a pan. All non-heated blends and cold-pressed oils were tested for fatty acid profile, content and composition of phytosterols, tocochromanols, chlorophyll and radical scavenging activity (RSA) analyzed by 2,2-diphenyl-1-picrylhydrazyl (DPPH), and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays. Moreover, the stability of phytosterols, tocochromanols, DPPH and ABTS values was determined in heated blends. All tocochromanols were lost during the heating process, in particular, at 200 °C. However, there were some differences between homologues. α-Tocopherol and δ-tocopherol were the most thermolabile and the most stable, respectively. Phytosterols were characterized by very high stability at both temperatures. We observed relationships between ABTS and DPPH values and contents of total tocochromanols and α-tocopherol. The obtained results may be useful in designing a new type of fried food with improved health properties and it may be the basis for further research on this topic.

Structural and Biochemical Characterization of a New Phage-Encoded Muramidase, KTN6 Gp46.

Background: Endolysins are phage-encoded lytic enzymes that degrade bacterial peptidoglycan at the end of phage lytic cycles to release new phage particles. These enzymes are being explored as an alternative to small-molecule antibiotics. Methods: The crystal structure of KTN6 Gp46 was determined and compared with a ColabFold model. Cleavage specificity was examined using a peptidoglycan digest and reversed-phase high-performance liquid chromatography coupled to mass spectrometry (HPLC/MS). Results: The structure of KTN6 Gp46 could be determined at 1.4 Å resolution, and key differences in loops of the putative peptidoglycan binding domain were identified in comparison with its closest known homologue, the endolysin of phage SPN1S. Reversed-phase HPLC/MS analysis of the reaction products following peptidoglycan digestion confirmed the muramidase activity of Gp46, consistent with structural predictions. Conclusion: These insights into the structure and function of endolysins further expand the toolbox for endolysin engineering and explore their potential in enzyme-based antibacterial design strategies.

Tomato Allergy: The Characterization of the Selected Allergens and Antioxidants of Tomato (Solanum lycopersicum)-A Review.

Tomatoes are one of the most broadly produced and consumed crop plants. They are the source of health-promoting nutrients such as antioxidants, including ascorbic acid, polyphenols, or carotenoids. Despite the beneficial role of tomatoes in the daily diet, they have been confirmed as one of the most prevalent allergenic vegetables. Food allergies can cause many clinical symptoms, e.g., in the gastrointestinal tract, skin, and lungs, as well as anaphylactic shock. A huge amount of clinical research has been carried out to improve the understanding of the immunological mechanisms that lead to the lack of tolerance of food antigens, which can result in either immunoglobulin E (IgE)-mediated reactions or non-IgE-mediated reactions. Lifestyle and diet play an important role in triggering food allergies. Allergy to tomatoes is also linked to other allergies, such as grass pollen and latex allergy. Numerous attempts have been made to identify and characterize tomato allergens; however, the data available on the subject are not sufficient.

Radical Scavenging Activity and Physicochemical Properties of Aquafaba-Based Mayonnaises and Their Functional Ingredients.

A plant-based diet has become more popular as a pathway to transition to more sustainable diets and personal health improvement in recent years. Hence, vegan mayonnaise can be proposed as an egg-free, allergy friendly vegan substitute for full-fat conventional mayonnaise. This study intends to evaluate the effect of aquafaba from chickpeas and blends of refined rapeseed oil (RRO) with different cold-pressed oils (10% of rapeseed oil-CPRO, sunflower oil-CPSO, linseed oil-CPLO or camelina oil-CPCO) on the radical scavenging, structural, emulsifying, and optical properties of novel vegan mayonnaise samples. Moreover, the functional properties and radical scavenging activity (RSA) of mayonnaise ingredients were evaluated. Aquafaba-based emulsions had a higher RSA than commercial vegan mayonnaise, determined by QUick, Easy, Novel, CHEap and Reproducible procedures using 2,2-diphenyl-1-picrylhydrazyl (QUENCHER-DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (QUENCHER-ABTS). Oxidative parameters such as peroxide values (PV), anisidine values (AnV), total oxidation (TOTOX) indexes and acid values (AV) of the proposed vegan mayonnaises were similar to those for commercial mayonnaises. Moreover, aquafaba-based samples had smaller oil droplet sizes than commercial vegan mayonnaise, which was observed using confocal laser scanning microscopy. The novel formulas developed in this study are promising alternatives to commercial vegan emulsions.

Sustainable or Not? Insights on the Consumption of Animal Products in Poland.

Animal products are one of the main constituents of the human diet. They are the main source of energy, proteins, microelements, and bioactive substances. The most popular negative health impacts linked with the consumption of animal products are obesity, atherosclerosis, heart attacks, and cancer. Apart from human health, consuming animal products is also controversial lately, due to farm animals' well-being and environmental protection issues. Thus, within the context of sustainability, the consumption trends of animal products were investigated through our on-line questionnaire surveys. The following animal products were involved in the survey: unprocessed meat (pork, beef, lamb, veal, mutton, chicken, duck, goose, turkey), processed meat (cold-cuts, sausages, pates), fish products, and eggs. Our research concluded that consumption among respondents with higher education was unsustainable for both unprocessed and processed meat, as eating habits in terms of type and quantity of consumed meat indicated respondents' unawareness. The consumption of fish products was also revealed as unsustainable regarding the quantity of fish consumed in terms of its beneficial nutritional values. Egg consumption was revealed as the most sustainable among the investigated animal products. However, insignificant egg consumption among the respondents showed the actual need of social education in terms of the current knowledge regarding the beneficial aspects of eggs.

Conventional or Organic? Motives and Trends in Polish Vegetable Consumption.

Vegetables constitute a major component of human food security. They are the main sources of essential nutrients including antioxidants, natural dyes, minerals, and vitamins. Eating habit issues related to the consumption of vegetables are gaining importance within the context of a healthy lifestyle, longevity, and physical fitness. Additionally, food quality is of primary importance, and so-called eco-food (defined as food as natural as possible, without fertilizers, pesticides, or preservatives) seems to be the most popular world-trend in healthy nutrition. Keeping these ideas in focus, research on vegetable consumption in Poland in the context of conventional or organic production was performed using online questionnaire surveys. The results revealed that the rate of vegetable consumption depended primarily on economic status, except for the potato, which was a staple cutting across all economic strata. Among the 108 analyzed respondents, 74% bought vegetables from certified organic farms. However, 59% bought organic vegetables "rarely" or "sometimes", and only 15% "often". Next, respondents chose to buy vegetables from fresh food markets (45%) and in local shops (41%). About 20% of the respondents acquired vegetables from their own farms. Among the reasons for choosing vegetables from certified organic farms, respondents mentioned in decreasing order: "desire for proper nutrition" (30%), "thinking that organic vegetables are healthier" (28%), and "organic vegetables are generally better" (7%).

A large TAT deletion in a tyrosinaemia type II patient.

A girl, born to unrelated Spanish parents, presented at 6 months of age with photophobia, keratitis, palmar hyperkeratosis and high plasma tyrosine levels, indicative of tyrosinaemia type II. Analysis of the tyrosine aminotransferase (TAT) gene revealed a paternally inherited frameshift mutation c.1213delCinsAG at codon 405 causing a premature stop codon, and a maternally inherited deletion of 193kb encompassing the complete TAT gene and three neighbouring genes. This is the first complete TAT deletion in tyrosinaemia type II described so far.

Susceptibility to Clotrimazole of Candida spp. Isolated from the Genitourinary System-A Single Center Study.

The aim of this study was to determine the susceptibility to clotrimazole of 125 isolates of Candida spp. originated from the genitourinary system of hospitalized patients as well as outpatients, tested in the mycological laboratory of Wroclaw Medical University in the years 1999-2018. The minimal inhibitory concentrations of clotrimazole and fluconazole were determined with the use of the microdilution method according to EUCAST, and the MFC was determined by subsequent subculture on Sabouraud agar. For the tested population of Candida yeasts, the MIC values of clotrimazole ranged from 0.008 to 8 mg/L, and MIC90 was 1 mg/L, whereas MIC50 was 0.008 mg/L. The minimal fungicidal concentration ranged between 1 and >8 mg/L. The great majority of the isolates (88%; 110/125) displayed MIC < 1 mg/L and were classified as WT (wild-type), whereas MIC ≥ 1 mg/L was determined for 2/61 (3.2%) isolates of C. albicans, 9/38 (23.6%) of C. glabrata, 1/2 of C. tropicalis, and 3/3 of C. guilliermondii. Six isolates (four of C. glabrata and two of C. albicans), defined as non-WT for clotrimazole, were classified as resistant to fluconazole, according to CBP from EUCAST. The isolates with elevated MIC to clotrimazole originated mostly from patients of the pediatric hematology unit, and their proportion in this population amounted to 17.8% (13 out of 73 isolates). In contrast, among strains from ambulatory patients, the highest observed MIC value was 1 mg/L (1 out of 37 isolates; 2.7%). The data obtained correlate well with those of most published studies on the in vitro susceptibility of Candida spp. to clotrimazole, which is usually very high. However, the existence of reports regarding the growing prevalence of resistant isolates has also to be noted. These results support the need for routinely checking the susceptibility of Candida clinical isolates to this imidazole derivative.