scholarly journals Combined Application of Bacteriophages and Carvacrol in the Control of Pseudomonas syringae pv. actinidiae Planktonic and Biofilm Forms

2020 ◽  
Vol 8 (6) ◽  
pp. 837
Author(s):  
Peien Ni ◽  
Lei Wang ◽  
Bohan Deng ◽  
Songtao Jiu ◽  
Chao Ma ◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Phage Therapy ◽  
Combined Treatment ◽  
Cost Effective ◽  
Resistance Development ◽  
Combined Application ◽  
Cost Effective Approach ◽  
Alternative Approach ◽  
Actinidia Spp

Pseudomonas syringae pv. actinidiae (Psa) is the causative agent of the bacterial canker of kiwifruit (Actinidia spp.). Phage therapy has been suggested as a viable alternative approach to controlling this disease, but its efficacy is limited by the emergence of phage-resistant mutants. Carvacrol is an essential oil that may be useful for the control of Psa. Combination therapies can be used to overcome resistance development. Here, the combination of phages (single phage suspensions of phages PN05 and PN09, and a cocktail of both phages) and carvacrol was investigated in controlling Psa planktonic and biofilm forms in vitro. The phage therapy alone (with phages PN05 and PN09), and the carvacrol alone (minimum inhibitory concentration 2.0 mg/mL), inhibited Psa growth, but the combined effect of both therapies was more effective. The phages alone effectively inhibited Psa growth for 24 h, but Psa regrowth was observed after this time. The carvacrol (2.0 mg/mL) alone prevented the biofilm formation for 48 h, but did not destroy the pre-formed biofilms. The combined treatment, phages and carvacrol (2.0 mg/mL), showed a higher efficacy, preventing Psa regrowth for more than 40 h. In conclusion, the combined treatment with phages and carvacrol may be a promising, environment-friendly and cost-effective approach to controlling Psa in the kiwifruit industry.

scholarly journals Pseudomonas aeruginosa PAO 1 In Vitro Time–Kill Kinetics Using Single Phages and Phage Formulations—Modulating Death, Adaptation, and Resistance

Antibiotics ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 877
Author(s):  
Ana Mafalda Pinto ◽  
Alberta Faustino ◽  
Lorenzo M. Pastrana ◽  
Manuel Bañobre-López ◽  
Sanna Sillankorva
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Phage Therapy ◽  
Multidrug Resistant ◽  
Resistant Bacteria ◽  
Chronic Infections ◽  
Swarming Motility ◽  
Resistance Development ◽  
Healthcare Settings ◽  
Time Kill

Pseudomonas aeruginosa is responsible for nosocomial and chronic infections in healthcare settings. The major challenge in treating P. aeruginosa-related diseases is its remarkable capacity for antibiotic resistance development. Bacteriophage (phage) therapy is regarded as a possible alternative that has, for years, attracted attention for fighting multidrug-resistant infections. In this work, we characterized five phages showing different lytic spectrums towards clinical isolates. Two of these phages were isolated from the Russian Microgen Sextaphage formulation and belong to the Phikmvviruses, while three Pbunaviruses were isolated from sewage. Different phage formulations for the treatment of P. aeruginosa PAO1 resulted in diversified time–kill outcomes. The best result was obtained with a formulation with all phages, prompting a lower frequency of resistant variants and considerable alterations in cell motility, resulting in a loss of 73.7% in swimming motility and a 79% change in swarming motility. These alterations diminished the virulence of the phage-resisting phenotypes but promoted their growth since most became insensitive to a single or even all phages. However, not all combinations drove to enhanced cell killings due to the competition and loss of receptors. This study highlights that more caution is needed when developing cocktail formulations to maximize phage therapy efficacy. Selecting phages for formulations should consider the emergence of phage-resistant bacteria and whether the formulations are intended for short-term or extended antibacterial application.

scholarly journals Protection of computers from side electromagnetic radiation during monitor images formation

2021 ◽  
Vol 4 (4) ◽  
pp. 377-385
Author(s):  
Volodymyr M. Lucenko ◽  
Dmytro O. Progonov
Keyword(s):  
Electromagnetic Radiation ◽  
Ad Hoc ◽  
Low Cost ◽  
Cost Effective ◽  
Switching Speed ◽  
Cost Effective Approach ◽  
Alternative Approach ◽  
Confidential Data ◽  
The Cost ◽  
Communication Lines

Reliable protection of confidential data processed in critical information infrastructure elements of public institutions and private organizations is topical task today. Of particular interest are methods to prevent the leakage of confidential data by localizing informative (dangerous) signals that both carry an informative component, and have a signal level higher than predefined threshold. The increase in signal energy from personal computers is caused by increasing of its transistors switching speed. Modern passive shielding methods for secured computers, similar to the well-known program TEMPEST, require either costly and large shielding units or technological simplification by using of low-cost fragmentary shielding of computer’s individual elements. Therefore, localization of side electromagnetic radiation produced by personal computer is needed. The paper presents a cost-effective approach to reducing the level of computer’s electromagnetic radiation by passive method. The radiation are localized and measured by its estimation on personal computer’s elements, namely unshielded communication lines between video processor and a monitor, fragments of electric tracks on motherboards, etc. During experiments authors used ad-hoc miniature electric (ball antenna) and magnetic (Hall sensor) antennas connected to selective voltmeters. This approach significantly reduces the cost of equipment and measurements as well as requirements to analytics’ qualification for improving computer’s protection. Also, the alternative approach for computer protection is proposed. The approach is based on image content protection by distorting the image on the monitor instead of reducing electromagnetic radiation caused by signals from the monitor. The protection includes image scrambling using Arnold transform that randomly “shuffle” the lines in each frame.

scholarly journals Rapid Identification of Potential Drug Candidates from Multi-Million Compounds’ Repositories. Combination of 2D Similarity Search with 3D Ligand/Structure Based Methods and In Vitro Screening

Molecules ◽  
2021 ◽  
Vol 26 (18) ◽  
pp. 5593
Author(s):  
Katalin Szilágyi ◽  
Beáta Flachner ◽  
István Hajdú ◽  
Mária Szaszkó ◽  
Krisztina Dobi ◽  
...  
Keyword(s):  
Similarity Search ◽  
Pharmacophore Model ◽  
Cost Effective ◽  
Rapid Identification ◽  
Drug Candidates ◽  
Hit Rate ◽  
Physico Chemical ◽  
Cost Effective Approach ◽  
3D Methods

Rapid in silico selection of target focused libraries from commercial repositories is an attractive and cost-effective approach in early drug discovery. If structures of active compounds are available, rapid 2D similarity search can be performed on multimillion compounds’ databases. This approach can be combined with physico-chemical parameter and diversity filtering, bioisosteric replacements, and fragment-based approaches for performing a first round biological screening. Our objectives were to investigate the combination of 2D similarity search with various 3D ligand and structure-based methods for hit expansion and validation, in order to increase the hit rate and novelty. In the present account, six case studies are described and the efficiency of mixing is evaluated. While sequentially combined 2D/3D similarity approach increases the hit rate significantly, sequential combination of 2D similarity with pharmacophore model or 3D docking enriched the resulting focused library with novel chemotypes. Parallel integrated approaches allowed the comparison of the various 2D and 3D methods and revealed that 2D similarity-based and 3D ligand and structure-based techniques are often complementary, and their combinations represent a powerful synergy. Finally, the lessons we learnt including the advantages and pitfalls of the described approaches are discussed.

scholarly journals Efficiency of Phage φ6 for Biocontrol of Pseudomonas syringae pv. syringae: An in Vitro Preliminary Study

2019 ◽  
Vol 7 (9) ◽  
pp. 286 ◽  
Author(s):  
Larindja A. M. Pinheiro ◽  
Carla Pereira ◽  
Carolina Frazão ◽  
Victor M. Balcão ◽  
Adelaide Almeida
Keyword(s):  
Solar Radiation ◽  
Host Range ◽  
Pseudomonas Syringae ◽  
Bacterial Resistance ◽  
Phage Therapy ◽  
Bacterial Species ◽  
Fruit Trees ◽  
Range Analysis ◽  
Maximum Reduction

Pseudomonas syringae is a plant-associated bacterial species that has been divided into more than 60 pathovars, with the Pseudomonas syringae pv. syringae being the main causative agent of diseases in a wide variety of fruit trees. The most common treatments for biocontrol of P. syringae pv. syringae infections has involved copper derivatives and/or antibiotics. However, these treatments should be avoided due to their high toxicity to the environment and promotion of bacterial resistance. Therefore, it is essential to search for new approaches for controlling P. syringae pv. syringae. Phage therapy can be a useful alternative tool to the conventional treatments to control P. syringae pv. syringae infections in plants. In the present study, the efficacy of bacteriophage (or phage) φ6 (a commercially available phage) was evaluated in the control of P. syringae pv. syringae. As the plants are exposed to the natural variability of physical and chemical parameters, the influence of pH, temperature, solar radiation and UV-B irradiation on phage φ6 viability was also evaluated in order to develop an effective phage therapy protocol. The host range analysis revealed that the phage, besides its host (P. syringae pv. syringae), also infects the Pseudomonas syringae pv. actinidiae CRA-FRU 12.54 and P. syringae pv. actinidiae CRA-FRU 14.10 strains, not infecting strains from the other tested species. Both multiplicities of infection (MOIs) tested, 1 and 100, were effective to inactivate the bacterium, but the MOI 1 (maximum reduction of 3.9 log CFU/mL) was more effective than MOI 100 (maximum reduction of 2.6 log CFU/mL). The viability of phage φ6 was mostly affected by exposure to UV-B irradiation (decrease of 7.3 log PFU/mL after 8 h), exposure to solar radiation (maximum reduction of 2.1 PFU/mL after 6 h), and high temperatures (decrease of 8.5 PFU/mL after 6 days at 37 °C, but a decrease of only 2.0 log PFU/mL after 67 days at 15 °C and 25 °C). The host range, high bacterial control and low rates of development of phage-resistant bacterial clones (1.20 × 10−3) suggest that this phage can be used to control P. syringae pv. syringae infections in plants, but also to control infections by P. syringae pv. actinidiae, the causal agent of bacterial canker of kiwifruit. Although the stability of phage φ6 was affected by UV-B and solar radiation, this can be overcome by the application of phage suspensions at the end of the day or at night.

scholarly journals Cytotoxic Effect of Combined Treatment with Ukrain and Methotrexate on Kidney Cells of Green Monkey

2012 ◽  
Vol 56 (1) ◽  
pp. 57-62
Author(s):  
Magdalena Izdebska ◽  
Iwona Piątkowska-Chmiel ◽  
Ewa Jagiełło-Wójtowicz
Keyword(s):  
Adverse Effect ◽  
Cell Culture ◽  
Mtt Assay ◽  
Cytotoxic Effect ◽  
Combined Treatment ◽  
Monkey Kidney ◽  
Kidney Cells ◽  
Combined Application ◽  
Green Monkey

Abstract In in vitro study on green monkey kidney (GMK) cell culture, the cytotoxicity of Ukrain, methotrexate (MTX), and their combination was investigated. The effect of these drugs and their combination on viability (MTT test) and apoptosis of the cells was assessed. The IC10 and IC50 values for Ukrain and MTX were also indicated. After 24 h of incubation of GMK cells with Ukrain, IC10 amounted to 84.6 μmol/L and IC50 was 256.3 μmol/L, while MTX IC10 amounted to 7.18 μmol/L and IC50 was 154.8 μmol/l. After 24 h of simultaneous incubation of GMK cells with 50 μmol/L of Ukrain and 5.5 μmol/L of MTX, 15.33 % of cytotoxicity of the drugs in LDH test was found. The most significant increase in the cytotoxicity (42.10 %) was noted after 24 h incubation of GMK cells with 150 μmol/L of Ukrain and 5.5 μmol/L of MTX. Likewise, in the MTT assay the greatest decrease in the cells viability was found after incubation with 150 μmol/L of Ukrain and 5.5 μmol/L of MTX. The evaluation of apoptosis also indicated the adverse effect of combined application of both drugs on GMK cells.

scholarly journals The utility of yeast as a tool for cell-based, target-directed high-throughput screening

Parasitology ◽  
2013 ◽  
Vol 141 (1) ◽  
pp. 8-16 ◽  
Author(s):  
J. L. NORCLIFFE ◽  
E. ALVAREZ-RUIZ ◽  
J. J. MARTIN-PLAZA ◽  
P. G. STEEL ◽  
P. W. DENNY
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Neglected Tropical Diseases ◽  
Cost Effective ◽  
Tropical Diseases ◽  
Protein Targets ◽  
Biochemical Assays ◽  
Cost Effective Approach ◽  
Cellular Context

SUMMARYMany Neglected Tropical Diseases (NTDs) have recently been subject of increased focus, particularly with relation to high-throughput screening (HTS) initiatives. These vital endeavours largely rely of two approaches, in vitro target-directed screening using biochemical assays or cell-based screening which takes no account of the target or targets being hit. Despite their successes both of these approaches have limitations; for example, the production of soluble protein and a lack of cellular context or the problems and expense of parasite cell culture. In addition, both can be challenging to miniaturize for ultra (u)HTS and expensive to utilize. Yeast-based systems offer a cost-effective approach to study and screen protein targets in a direct-directed manner within a eukaryotic cellular context. In this review, we examine the utility and limitations of yeast cell-based, target-directed screening. In particular we focus on the currently under-explored possibility of using such formats in uHTS screening campaigns for NTDs.

scholarly journals Effect of once weekly oral levothyroxine therapy

2021 ◽  
Vol 2021 ◽  
Author(s):  
Andrea del Toro-Diez ◽  
Ernesto Solá-Sánchez ◽  
Michelle Mangual-García
Keyword(s):  
Case Series ◽  
Cost Effective ◽  
Primary Hypothyroidism ◽  
Endocrine Disorders ◽  
List Type ◽  
Common Cause ◽  
Cost Effective Approach ◽  
Alternative Approach ◽  
Learning Points ◽  
Tsh Levels

Summary Primary hypothyroidism is one of the most common endocrine disorders with widely available treatment. A minority of patients remain with uncontrolled hypothyroidism despite therapy. The objective of this case series was to demonstrate that medication non-adherence, rather than malabsorption, should be sought as the most common cause of unsuppressed TSH levels in patients receiving treatment for this condition. Non-adherence is often considered as a diagnosis of exclusion. Nonetheless, a diagnosis of malabsorption requires a more extensive workup, including imaging and invasive procedures, which increase healthcare costs and burden to the patient. The findings of this study allow for a cost-effective approach to uncontrolled hypothyroidism. Learning points Medication non-adherence is a common cause of insuppressible TSH levels. Once weekly levothyroxine is an alternative approach to non-compliant patients. Assessing compliance is more cost-effective and less burdensome than testing for malabsorption.

scholarly journals Cell Electropermeabilisation Enhancement by Non-Thermal-Plasma-Treated PBS

Cancers ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 219 ◽  
Author(s):  
Thai-Hoa Chung ◽  
Augusto Stancampiano ◽  
Kyriakos Sklias ◽  
Kristaq Gazeli ◽  
Franck André ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Electric Fields ◽  
Combined Treatment ◽  
Chinese Hamster ◽  
Field Amplitude ◽  
Lung Fibroblasts ◽  
Cancer Therapies ◽  
Electric Pulses ◽  
Combined Application

The effectiveness of electrochemotherapy (ECT) in local eradication of tumours in human and veterinary medicine has been proven. ECT consists of increasing the uptake of cytotoxic drugs by means of pulsed electric fields (PEFs) that transiently permeabilise the cell membrane. Still, this tumour treatment includes some drawbacks that are linked to the characteristics of the intense electric pulses (EPs) used. Meanwhile, the emerging field of cancer therapies that are based on the application of non-thermal plasmas (NTP) has recently garnered interest because of their potentialities as rich sources of reactive species. In this work, we investigated the potential capabilities of the combined application of indirect NTP treatment and microsecond PEFs (µsPEFs) to outperform in vitro cell electropermeabilisation, the basis of ECT. Thus, phosphate-buffered saline (PBS) was plasma-treated (pPBS) and used afterwards to explore the effects of its combination with µsPEFs. Analysis of two different cell lines (DC-3F Chinese hamster lung fibroblasts and malignant B16-F10 murine melanoma cells), by flow cytometry, revealed that this combination resulted in significant increases of the level of cell membrane electropermeabilisation, even at very low electric field amplitude. The B16-F10 cells were more sensitive to the combined treatment than DC-3F cells. Importantly, the percentage of permeabilised cells reached values similar to those of cells exposed to classical electroporation field amplitude (1100 V/cm) when the cells were treated with pPBS before and after being exposed only to very low PEF amplitude (600 V/cm). Although the level of permeabilisation of the cells that are treated by the pPBS and the PEFs at 600 V/cm is lower than the level reached after the exposure to µsPEFs alone at 1100 V/cm, the combined treatment opens the possibility to reduce the amplitude of the EPs used in ECT, potentially allowing for a novel ECT with reduced side-effects.

scholarly journals Characterization of a Lytic Bacteriophage against Pseudomonas syringae pv. actinidiae and Its Endolysin

Viruses ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 631
Author(s):  
Peien Ni ◽  
Lei Wang ◽  
Bohan Deng ◽  
Songtao Jiu ◽  
Chao Ma ◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Phage Therapy ◽  
Gc Content ◽  
Lytic Activity ◽  
Lytic Bacteriophage ◽  
Double Stranded Dna ◽  
Alternative Approaches ◽  
Myoviridae Family

Pseudomonas syringae pv. actinidiae (Psa) is a phytopathogen that causes canker in kiwifruit. Few conventional control methods are effective against this bacterium. Therefore, alternative approaches, such as phage therapy are warranted. In this study, a lytic bacteriophage (PN09) of Psa was isolated from surface water collected from a river in Hangzhou, China in 2019. Morphologically, PN09 was classified into the Myoviridae family, and could lyse all 29 Psa biovar 3 strains. The optimal temperature and pH ranges for PN09 activity were determined as 25 to 35 ∘C and 6.0 to 9.0, respectively. The complete genome of PN09 was found to be composed of a linear 99,229 bp double-stranded DNA genome with a GC content of 48.16%. The PN09 endolysin (LysPN09) was expressed in vitro and characterized. LysPN09 was predicted to belong to the Muraidase superfamily domain and showed lytic activity against the outer-membrane-permeabilized Psa strains. The lytic activity of LysPN09 was optimal over temperature and pH ranges of 25 to 40 ∘C and 6.0 to 8.0, respectively. When recombinant endolysin LysPN09 was combined with EDTA, Psa strains were effectively damaged. All these characteristics demonstrate that the phage PN09 and its endolysin, LysPN09, are potential candidates for biocontrol of Psa in the kiwifruit industry.

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