scholarly journals THE FITNESS OF A RECOMBINANT STRAIN OF PSEUDOMONAS AERUGINOSA BACTERIA COMPARING WITH ITS PARENT STRAINS UNDER THE EGYPTIAN ENVIRONMENTAL CONDITIONS (MOWAS RIVER) IN 2020

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Salah E. Maaty ◽  
Ahmed Sh. Hassan ◽  
Mamdouh K. Amin ◽  
Mohamed M.A. Elashtokhy
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Recombinant Strain ◽  
Genetically Modified ◽  
Genetic Material ◽  
Genetically Engineered ◽  
Genetically Modified Microorganisms ◽  
Bacterial Fitness ◽  
Environmental Fitness

The present study aimed to study the fitness between a trans-conjugant (recombinant strain) of Pseudomonas aeruginosa bacteria with its parents after transferring genetic material by conjugation mechanism. Whereas, environmental fitness expresses the interaction of an organism with its environment therefore it is considered a good indicator for the assessment of genetically engineered microorganisms (GEM) released into nature. Incubation time was carried out in vitro and incubating time in situ in Mowas River Zagazig city during winter and summer. Accordingly, the fitness of the parents and the recombinant strain was studied. The three strains of Pseudomonas aeruginosa (PAO1, MAM2 and PU21) were tested on chloramphenicol and tetracycline. Strain MAM2 was resistant to chloramphenicol 1200 µg/ml while was sensitive to tetracycline and has been used as the recipient. While strain PAO1 was resistant to tetracycline 200 µg/ml and was sensitive to chloramphenicol and has been used as the donor. Results proofed the presence of the plasmid in the donor and trans conjugant strains. The donor was treated with acridine orange to match the results obtained with the results at the molecular level. It was observed that bacterial fitness continued for up to 35 days in vitro, while in situ during the summer it did not last at the site for only 21 days. While it lasted 28 days during the summer. So, the risks that may be caused by releasing the genetically modified microorganisms into environments have been canceled. In addition to its ability to preserve the new genetic material, it may be able to transfer this new genetic material to other strains and species that may be live in the same ecosystem, as it is largely stable in the environment. In genetically modified microorganisms that are added to the environments for agricultural uses such as increasing soil fertility (bio-fertilizer) or biodegradation for a harmful substance such as pesticides, the soil must be re-inoculated in

The Health Effects of Genetically Modified Foods: a Brief Review

2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Chalachew Chekol ◽  
Keyword(s):  
Poverty Reduction ◽  
Deoxyribonucleic Acid ◽  
Genetically Modified ◽  
New Technology ◽  
Genetic Material ◽  
Genetically Engineered ◽  
Environmental Conservation ◽  
Genetically Modified Foods ◽  
Weather Extremes ◽  
Recombinant Technology

Genetically modified foods are organisms (i.e. plants or animals) in which the genetic material (Deoxyribonucleic Acid) has been altered in a way that does not occur naturally by mating and/or natural recombination. Combining genes from different organisms is known as recombinant Deoxyribonucleic Acid technology and the resulting organism is said to be ‘Genetically Modified’, ‘Genetically Engineered’ or ‘Transgenic’. Crops grown commercially and/or field-tested are resistant to a virus that could destroy most of the African harvest, other crops with increased iron and vitamins that may alleviate chronic malnutrition and a variety of plants that are able to survive weather extremes. There are fruits that produce human vaccines against infectious diseases such as hepatitis B, fish that mature more quickly, fruit and nut trees that yield years earlier and plants that produce new plastics with unique properties. Controversies and public concern surrounding Genetically Modified foods and crops commonly focus on human and environmental safety, ethics, food security, poverty reduction and environmental conservation. With this new technology on gene manipulation there are the risks of tampering with nature, effects will have on the environment, the health concerns that consumers should be aware of, and effects related with recombinant technology. This review addresses the major concerns about the safety, environmental and legal issues which are collectively infer health hazards of Genetically Modified foods and recombinant technology in different perspective.

scholarly journals Small-Molecule Inhibition of Choline Catabolism in Pseudomonas aeruginosa and Other Aerobic Choline-Catabolizing Bacteria

2011 ◽  
Vol 77 (13) ◽  
pp. 4383-4389 ◽  
Author(s):  
Liam F. Fitzsimmons ◽  
Stevenson Flemer ◽  
A. Sandy Wurthmann ◽  
P. Bruce Deker ◽  
Indra Neil Sarkar ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Burkholderia Cepacia ◽  
Sinorhizobium Meliloti ◽  
Growth Inhibitor ◽  
Bioinformatic Analysis ◽  
Content Type ◽  
Genes Encoding ◽  
Dependent Growth

ABSTRACTCholine is abundant in association with eukaryotes and plays roles in osmoprotection, thermoprotection, and membrane biosynthesis in many bacteria. Aerobic catabolism of choline is widespread among soil proteobacteria, particularly those associated with eukaryotes. Catabolism of choline as a carbon, nitrogen, and/or energy source may play important roles in association with eukaryotes, including pathogenesis, symbioses, and nutrient cycling. We sought to generate choline analogues to study bacterial choline catabolismin vitroandin situ. Here we report the characterization of a choline analogue, propargylcholine, which inhibits choline catabolism at the level of Dgc enzyme-catalyzed dimethylglycine demethylation inPseudomonas aeruginosa. We used genetic analyses and13C nuclear magnetic resonance to demonstrate that propargylcholine is catabolized to its inhibitory form, propargylmethylglycine. Chemically synthesized propargylmethylglycine was also an inhibitor of growth on choline. Bioinformatic analysis suggests that there are genes encoding DgcA homologues in a variety of proteobacteria. We examined the broader utility of propargylcholine and propargylmethylglycine by assessing growth of other members of the proteobacteria that are known to grow on choline and possess putative DgcA homologues. Propargylcholine showed utility as a growth inhibitor inP. aeruginosabut did not inhibit growth in other proteobacteria tested. In contrast, propargylmethylglycine was able to inhibit choline-dependent growth in all tested proteobacteria, includingPseudomonas mendocina,Pseudomonas fluorescens,Pseudomonas putida,Burkholderia cepacia,Burkholderia ambifaria, andSinorhizobium meliloti. We predict that chemical inhibitors of choline catabolism will be useful for studying this pathway in clinical and environmental isolates and could be a useful tool to study proteobacterial choline catabolismin situ.

scholarly journals Genetically edited CD34+ cells derived from human iPS cells in vivo but not in vitro engraft and differentiate into HIV-resistant cells

2021 ◽  
Vol 118 (20) ◽  
pp. e2102404118
Author(s):  
Maelig G. Morvan ◽  
Fernando Teque ◽  
Lin Ye ◽  
Mary E. Moreno ◽  
Jiaming Wang ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Genetically Modified ◽  
Ips Cells ◽  
Genetically Engineered ◽  
Hiv Cure ◽  
Peripheral Blood Mononuclear ◽  
Hematopoietic Stem ◽  
Immunodeficient Mice

Genetic editing of induced pluripotent stem (iPS) cells represents a promising avenue for an HIV cure. However, certain challenges remain before bringing this approach to the clinic. Among them, in vivo engraftment of cells genetically edited in vitro needs to be achieved. In this study, CD34+ cells derived in vitro from iPS cells genetically modified to carry the CCR5Δ32 mutant alleles did not engraft in humanized immunodeficient mice. However, the CD34+ cells isolated from teratomas generated in vivo from these genetically edited iPS cells engrafted in all experiments. These CD34+ cells also gave rise to peripheral blood mononuclear cells in the mice that, when inoculated with HIV in cell culture, were resistant to HIV R5-tropic isolates. This study indicates that teratomas can provide an environment that can help evaluate the engraftment potential of CD34+ cells derived from the genetically modified iPS cells in vitro. The results further confirm the possibility of using genetically engineered iPS cells to derive engraftable hematopoietic stem cells resistant to HIV as an approach toward an HIV cure.

Autologous, Orthotopic Thyroid Follicular Cell Transplantation: A Surgical Component of Ex Vivo Somatic Gene Therapy

1993 ◽  
Vol 108 (1) ◽  
pp. 51-62 ◽  
Author(s):  
Bert W. O'Malley ◽  
Milton J. Finegold ◽  
Fred D. Ledley
Keyword(s):  
Gene Therapy ◽  
Ex Vivo ◽  
Genetic Material ◽  
Genetically Engineered ◽  
Follicular Cells ◽  
Somatic Gene Therapy ◽  
Orthotopic Transplantation ◽  
Somatic Gene ◽  
Thyroid Follicular Cells

Ex vivo strategies for somatic gene therapy involve the harvest of primary cells from patients, the transfer of novel genetic material into these cells in cell culture, and reimplantation of the genetically engineered cells back into patients. In consideration of methods for targeting somatic gene therapy to the thyroid, we have studied the autologous, orthotopic transplantation of thyroid follicular cells in a canine model. Using the fluorescent dye Dil, we were able to stain follicular cells in vitro before transplantation and then follow the pattern of engraftment through histologic sectioning and microscopy up to 14 days after transplantation. The transplantations involved the direct injection of intact and disrupted follicles into a remaining thyroid lobe after cell harvest from the contralateral lobe. We also demonstrated engraftment of individual follicular cells recovered from primary monolayer cultures. Histologic studies revealed the presence of transplanted cells and follicles as well as focal regions of granulomatous reaction in close relation to the engrafted material. These studies demonstrate the feasibility of autologous, orthotopic transplantation of thyroid follicular cells. This method is an essential component of ex vivo strategies for targeting somatic gene therapy to the thyroid gland.

scholarly journals Optimal environmental and culture conditions allow the in vitro coexistence of Pseudomonas aeruginosa and Staphylococcus aureus in stable biofilms

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Maria del Mar Cendra ◽  
Núria Blanco-Cabra ◽  
Lucas Pedraz ◽  
Eduard Torrents
Keyword(s):  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Culture Media ◽  
Culture Conditions ◽  
Single Organism ◽  
Bacterial Fitness ◽  
Major Pathogens ◽  
And Staphylococcus Aureus ◽  
Stable Formation

Abstract The coexistence between species that occurs in some infections remains hard to achieve in vitro since bacterial fitness differences eventually lead to a single organism dominating the mixed culture. Pseudomonas aeruginosa and Staphylococcus aureus are major pathogens found growing together in biofilms in disease-affected lungs or wounds. Herein, we tested and analyzed different culture media, additives and environmental conditions to support P. aeruginosa and S. aureus coexistence in vitro. We have unraveled the potential of DMEM to support the growth of these two organisms in mature cocultured biofilms (three days old) in an environment that dampens the pH rise. Our conditions use equal initial inoculation ratios of both strains and allow the stable formation of separate S. aureus microcolonies that grow embedded in a P. aeruginosa biofilm, as well as S. aureus biofilm overgrowth when bovine serum albumin is added to the system. Remarkably, we also found that S. aureus survival is strictly dependent on a well-characterized phenomenon of oxygen stratification present in the coculture biofilm. An analysis of differential tolerance to gentamicin and ciprofloxacin treatment, depending on whether P. aeruginosa and S. aureus were growing in mono- or coculture biofilms, was used to validate our in vitro coculture conditions.

The production of antibacterial tubing, sutures, and bandages by in situ precipitation of metallic salts

1991 ◽  
Vol 37 (6) ◽  
pp. 445-449 ◽  
Author(s):  
Samuel R. Farrah ◽  
Gregory W. Erdos
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Silver Ions ◽  
Silicone Tubing ◽  
Metallic Salts ◽  
Second Procedure ◽  
And Staphylococcus Aureus

Two procedures were used to modify gauze bandages, polyester sutures, silicone tubing, and polyvinyl chloride tubing. In one procedure, the materials were first modified by in situ precipitation of metallic hydroxides and then used to adsorb silver ions. In the second procedure, the materials were soaked in sodium pyrophosphate or sodium chloride, dried, and then soaked in silver nitrate. These procedures produced materials with silver deposited on the surface of the tubing and sutures and both on the surface and within the gauze fibers. The modified materials inhibited the growth of Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus in vitro. Key words: tubing, sutures, bandages, antibacterial.

scholarly journals In situ Intranasal Delivery Systems: Application Prospects and Main Pharmaceutical Aspects of Development (Review)

2021 ◽  
Vol 10 (4) ◽  
pp. 54-63
Author(s):  
E. O. Bakhrushina ◽  
N. B. Demina ◽  
M. M. Shumkova ◽  
P. S. Rodyuk ◽  
D. S. Shulikina ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Delivery Systems ◽  
Genetically Engineered ◽  
Biological Molecules ◽  
Poloxamer 407 ◽  
Prolonged Release ◽  
Preclinical Studies ◽  
Intranasal Delivery

Introduction. Intranasal delivery of in situ gel-forming systems is a complex but promising direction. Due to the high cost of developing a new chemical object or genetically engineered modification of biological molecules, pharmaceutical companies are focusing on developing technologies for new delivery systems for existing active pharmaceutical ingredients to improve their effectiveness and bioavailability. In situ systems for intranasal delivery, due to increased viscosity and mucoadhesion to the nasal mucosa, allow overcoming mucociliary clearance and ensuring complete absorption and prolonged release of drugs.Text. The article discusses the main advantages of intranasal in situ delivery systems shown in preclinical studies, as well as approaches to the technology of obtaining and standardization of these systems. The results of scientific research in this field over the past 15 years are summarized, the most promising polymers for creating thermoreversible and pH-sensitive compositions are identified, and modern methods for evaluating the sol-gel transition in situ are analyzed.Conclusion. The use of in situ systems for intranasal administration allows providing a high targeting of the delivery of synthetic and biological molecules to the brain. Currently, numerous pharmacokinetic and pharmacodynamic preclinical studies confirm the effectiveness of such systems, as well as their safety. Thermoreversible commercially available and directionally synthesized polymers (poloxamer 407, PLGA, NIPAAm, etc.), as well as chitosan, remain the most popular for the design of in situ delivery systems. In vitro and ex vivo methods with mucosa and artificial nasal fluid are widely used to assess the parameters of in situ gelation, but to increase the reproducibility of the methods and improve the correlation in vitro/in vivo, it is recommended to conduct modeling of the nasal cavity. Developing the technology and methods of screening of intranasal reversible systems will help to get closer to clinical trials and the entry of these delivery systems into the global pharmaceutical market.

In Vivo and in Vitro Degradation of Urea and Uric Acid by Encapsulated Genetically Modified Microorganisms

2004 ◽  
Vol 10 (9) ◽  
pp. 1446-1455
Author(s):  
Jill A. O'Loughlin ◽  
Jan M. Bruder ◽  
Michael J. Lysaght
Keyword(s):  
Uric Acid ◽  
Genetically Modified ◽  
Genetically Modified Microorganisms
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