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Biomedical Physics & Engineering Express Feb 2023Fluid shear stress (FSS) is an important parameter that regulates various cell functions such as proliferation and migration. While there are a number of techniques to...
Fluid shear stress (FSS) is an important parameter that regulates various cell functions such as proliferation and migration. While there are a number of techniques to generate FSS, many of them require physical deformation or movement of solid objects to generate the fluid shear, making it difficult to decouple the effects of FSS and mechanical strains. This work describes the development of a non-mechanical means to generate fluid flow and FSS in a 2Dsetting. This was accomplished with a magnetohydrodynamic (MHD) pump, which creates liquid flow by generating a Lorentz force through the interaction between an electric field and an orthogonal magnetic field. The MHD pump system presented here consisted of trapezoidal prism-shaped magnets, a pair of platinum electrodes, and a modified petri dish. The system was validated and tested on anwound model, which is based on analyzing the migration of fibroblast cells through an artificially created scratch on a confluent cell culture surface. Experiments were performed to a control group, an electric field only group, and a group that was subject to fluid flow with the application of both electric field and magnetic field. Results show that fibroblast cells that experienced fluid shear have higher wound closure rate compared to the control group and the electric field only group. The data shows that the MHD pump can be a great tool to study FSS. Furthermore, due to its fluid flow generation without mechanical force, this system can be adapted and implemented to study the role of FSS and electric field on wound healing.
Topics: Cell Movement; Stress, Mechanical
PubMed: 36716480
DOI: 10.1088/2057-1976/acb711 -
Lab on a Chip Feb 2021The selection of high quality sperm is critical for intracytoplasmic sperm injection (ICSI), a prevalent assisted reproduction technology. However, standard selection...
The selection of high quality sperm is critical for intracytoplasmic sperm injection (ICSI), a prevalent assisted reproduction technology. However, standard selection methods are time-consuming and fail to recover the most viable sperm, thereby limiting the ICSI success rate. Microfluidics enables rapid selection of viable sperm in a manner representing in vivo processes, however, existing platforms lack clinical applicability. Here, we present FertDish, which integrates the clinically established ICSI Petri dish with a film featuring an array of sperm-selecting microchannels for selection of sperm directly from semen. The FertDish format mimics the clinician-familiar ICSI dish setup, and provides rapid (<10 min) single stage sperm preparation that circumvents standard labour-intensive multi-stage sperm processing steps. Tests with human donor and patient semen samples show that FertDish enables the selection of a high quality sperm sub-population, featuring improvements in DNA fragmentation index of more than 91% (donor) and 74% (patient) versus raw semen and 50% (donor) and 63% (patient) versus standard methods, and a distribution of more than 97% sperm with viable and high level DNA. The FertDish enables a high sperm recovery rate (>3.3 × 105 sperm per mL), and is readily adaptable to the clinical workflow with potential to improve ICSI outcomes.
Topics: Humans; Male; Microfluidics; Sperm Injections, Intracytoplasmic; Spermatozoa
PubMed: 33507191
DOI: 10.1039/d0lc00874e -
HardwareX Apr 2022Multispectral imaging is at the forefront of contactless surface analysis. Standard multispectral imaging systems use sophisticated software, cameras and light filtering...
Multispectral imaging is at the forefront of contactless surface analysis. Standard multispectral imaging systems use sophisticated software, cameras and light filtering optics. This paper discloses the building of a customizable and cost-effective multispectral imaging and analysis system. It integrates a web camera, light emitting diodes (LEDs) lighting, a semisphere for even lightening, an open-source Arduino™ development board and a free Python application to automatically obtain and visually analyze multispectral images. The device is hereafter called MEDUSA and its optical performance was tested for repeated Imaging consistency, visible and near infrared band sensitivity and lighting evenness. Four proof of concept tests were run in order to understand the advantageous use of this system, as compared to a simple visual score of diverse samples. Each of three qualitative tests used sets of 12 LED band spectral images to analyze ink changes in a counterfeit bill, surface bruises on Hass avocado fruits and transient changes in petri dish grown bacterial colonies. A fourth test used single band imaging in a set of standard laboratory analyzed plant samples, to quantitatively relate a red band light reflectance to its nitrogen content. These tests indicate that MEDUSA made images may yield qualitative and quantitative spectral information unseen to the naked eye, suggesting potential use in currency counterfeit tests, food quality analyses, microbial phenotyping and agricultural plant chemistry. MEDUSA can be freely reproduced and customized from this research, making it a powerful and affordable analytical tool to analyze a wide range of subtle chemical properties in samples at industrial and science fields.
PubMed: 35509904
DOI: 10.1016/j.ohx.2022.e00282 -
Environmental Science and Pollution... Nov 2023The possibility of using the non-nitrogen-fixing cyanobacterium (Chroococcus sp.) for the reduction of soil nitrate contamination was tested through Petri dish...
The possibility of using the non-nitrogen-fixing cyanobacterium (Chroococcus sp.) for the reduction of soil nitrate contamination was tested through Petri dish experiments. The application of 0.03, 0.05 and 0.08 mg/cm Chroococcus sp. efficiently removed NO-N from the soil through assimilation of nitrate nutrient and promotion of soil denitrification. At the optimal application dose of 0.05 mg/cm, 44.06%, 36.89% and 36.17% of NO-N were removed at initial NO-N concentrations of 60, 90 and 120 mg/kg, respectively. The polysaccharides released by Chroococcus sp. acted as carbon sources for bacterial denitrification and facilitated the reduction of soil salinity, which significantly (p < 0.05) stimulated the growth of denitrifying bacteria (Hyphomicrobium denitrificans and Hyphomicrobium sp.) as well as significantly (p < 0.05) elevated the activities of nitrate reductase and nitrite reductase by 1.07-1.23 and 1.15-1.22 times, respectively. The application of Chroococcus sp. promoted the dominance of Nocardioides maradonensis in soil microbial community, which resulted in elevated phosphatase activity and increased available phosphorus content. The application of Chroococcus sp. positively regulated the growth of soil bacteria belonging to the genera Chitinophaga, Prevotella and Tumebacillus, which may contribute to increased soil fertility through the production of beneficial enzymes such as invertase, urease and catalase. To date, this is the first study verifying the remediation effect of non-nitrogen-fixing cyanobacteria on nitrate-contaminated soil.
Topics: Nitrates; Cyanobacteria; Nitrate Reductase; Nitrite Reductases; Soil; Denitrification
PubMed: 37870669
DOI: 10.1007/s11356-023-30383-1 -
ELife Jan 2020Diverse interactions among species within bacterial colonies lead to intricate spatiotemporal dynamics, which can affect their growth and survival. Here, we describe the...
Diverse interactions among species within bacterial colonies lead to intricate spatiotemporal dynamics, which can affect their growth and survival. Here, we describe the emergence of complex structures in a colony grown from mixtures of motile and non-motile bacterial species on a soft agar surface. Time-lapse imaging shows that non-motile bacteria 'hitchhike' on the motile bacteria as the latter migrate outward. The non-motile bacteria accumulate at the boundary of the colony and trigger an instability that leaves behind striking flower-like patterns. The mechanism of the front instability governing this pattern formation is elucidated by a mathematical model for the frictional motion of the colony interface, with friction depending on the local concentration of the non-motile species. A more elaborate two-dimensional phase-field model that explicitly accounts for the interplay between growth, mechanical stress from the motile species, and friction provided by the non-motile species, fully reproduces the observed flower-like patterns.
Topics: Acinetobacter; Escherichia coli; Friction; Microbial Interactions; Models, Biological; Movement; Stress, Mechanical
PubMed: 31933477
DOI: 10.7554/eLife.48885 -
Frontiers in Plant Science 2022Coumestrol (CMS) derivatives are unique compounds, which function as phytoalexins; they are derived from soybean roots, following abiotic and biotic stresses. As a...
Coumestrol (CMS) derivatives are unique compounds, which function as phytoalexins; they are derived from soybean roots, following abiotic and biotic stresses. As a phytoalexin, CMS forms a defense system that enables plants to maintain their viability. However, it is still challenging to achieve the mass production of phytoalexins, which exhibit pharmacological values, plant breeding. Here, the synthesis of CMS derivatives from the seedling, plant, and adventitious root (AR) of were investigated under artificial light, as well as a chemical elicitor treatment. In the presence of constant light, as well as under treatment with methyl jasmonate, the CMS monoglucoside (coumestrin; CMSN) and malonyl CMSN (M-CMSN) contents of the AR culture (4 weeks) increased drastically. The two CMS derivatives, CMSN and M-CMSN, were obtained as a mixture of isomers, which were identified nuclear magnetic resonance analysis. These derivatives were also observed in a soybean plant that was grown on artificial soil (AS; 5 weeks) and a Petri dish (9 days) although in considerably lesser quantities than those observed in the AR culture. Compared with the two other media (AS and the Petri dish), the AR culture achieved the superior synthesis of CMSN and M-CMSN within a relatively short cultivation period (<1 month) in laboratory-scale (3 L) and pilot-scale (1,000 L) bioreactors. The isoflavone content of AR under the constant light conditions was three-fold that under dark conditions. Significant quantities of malonyl daidzin and malonyl genistin were produced in the root of AS and the seedling of Petri dish, respectively. Flavonol glycosides were not produced in the AR culture under the dark and light conditions, as well as in AS under the dark condition. However, significant contents of kaempferol glycosides were produced in the leaves of AS and seedling of Petri dish, following the light treatment. Thus, we proposed that the established soybean AR-cultivation approach represented a better method for biosynthesizing phytoalexins, such as the CMS derivatives, as plant-derived functional materials.
PubMed: 35800610
DOI: 10.3389/fpls.2022.923163 -
Frontiers in Microbiology 2021Cocultivation is an emerging and potential way to investigate microbial interaction in the laboratory. Extensive researches has been carried out over the years, but some...
Cocultivation is an emerging and potential way to investigate microbial interaction in the laboratory. Extensive researches has been carried out over the years, but some microorganism cocultivation are not easy to implement in the laboratory, especially the fungus-fungus (FF) cocultivation, owing to the obstacles such as fungal different growth rate, limited growing space, hyphae intertwining, and difficulty of sample separation, etc. In this research, a double-sided petri dish (DSPD) was designed and carried out as a tool to study FF cocultivation in the laboratory. A natural FF cocultivation of spp. and inspired from black-skin-red-koji (BSRK), were studied. By using DSPD, the aforementioned obstacles in the FF cocultivation study were overcome through co-culturing spp. and on each side of DSPD. The characteristics of monocultured and co-cultured spp. and were compared and analyzed, including colonial and microscopic morphologies, and main secondary metabolites (SMs) of spp. analyzed by high performance liquid chromatography. And a novel SM was found to be produced by M7 when co-cultured with CBS 513.88. Since the above mentioned obstacles, were overcome, we obtained good quality of transcriptome data for further analysis. These results indicate that DSPD might be an efficient tool for investigation of microbial interaction, in particular, for FF interaction.
PubMed: 34177849
DOI: 10.3389/fmicb.2021.670684 -
Plants (Basel, Switzerland) Nov 2023Oats () hold immense economic and nutritional value as a versatile crop. They have long been recognized as an exceptional choice for human consumption and animal feed.... (Review)
Review
Oats () hold immense economic and nutritional value as a versatile crop. They have long been recognized as an exceptional choice for human consumption and animal feed. Oats' unique components, including proteins, starches, and β-glucans, have led to its widespread use in various food products such as bread, noodles, flakes, and milk. The popularity of oat milk as a vegan alternative to dairy milk has soared due to the increasing number of vegetarians/vegans and growing environmental awareness. Oat milk offers a sustainable option with reduced greenhouse gas emissions during its production, rendering it an appropriate choice for individuals who are lactose-intolerant or have dairy allergies. To ensure improved adaptability and enhanced nutrition, the development of new oat varieties is crucial, considering factors like cultivation, climate, and growing conditions. Plant cell culture plays a crucial role in both traditional and contemporary breeding methods. In classical breeding, plant cell culture facilitates the rapid production of double haploid plants, which can be employed to accelerate the breeding process. In modern breeding methods, it enables genetic manipulation and precise genome editing at the cellular level. This review delves into the importance of oats and their diverse applications, highlighting the advantages of plant cell culture in both classical and modern breeding methods. Specifically, it provides an overview of plant tissue culture, encompassing genetic transformation, haploid technology, protoplast technology, and genome editing.
PubMed: 37960138
DOI: 10.3390/plants12213782 -
Angewandte Chemie (International Ed. in... Oct 2019Playing with evolution: In his Nobel lecture, George P. Smith reconstructs the story of the phage-display idea as he personally experienced it. The development of this... (Review)
Review
Playing with evolution: In his Nobel lecture, George P. Smith reconstructs the story of the phage-display idea as he personally experienced it. The development of this technique is a case study in how a scientific advance emerges gradually in incremental steps within overlapping global scientific communities.
Topics: Bacteriophages; Cell Surface Display Techniques; DNA, Viral; Evolution, Molecular; Nobel Prize; Peptide Library
PubMed: 31529666
DOI: 10.1002/anie.201908308 -
Journal of Visualized Experiments : JoVE Oct 2019One of the main challenges in the search for new antibiotics from natural product extracts is the re-discovery of common compounds. To address this challenge,...
One of the main challenges in the search for new antibiotics from natural product extracts is the re-discovery of common compounds. To address this challenge, dereplication, which is the process of identifying known compounds, is performed on samples of interest. Methods for dereplication such as analytical separation followed by mass spectrometry are time-consuming and resource-intensive. To improve the dereplication process, we have developed the antibiotic resistance platform (ARP). The ARP is a library of approximately 100 antibiotic resistance genes that have been individually cloned into Escherichia coli. This strain collection has many applications, including a cost-effective and facile method for antibiotic dereplication. The process involves the fermentation of antibiotic-producing microbes on the surface of rectangular Petri dishes containing solid medium, thereby allowing for the secretion and diffusion of secondary metabolites through the medium. After a 6 day fermentation period, the microbial biomass is removed, and a thin agar-overlay is added to the Petri dish to create a smooth surface and enable the growth of the E. coli indicator strains. Our collection of ARP strains is then pinned onto the surface of the antibiotic-containing Petri dish. The plate is next incubated overnight to allow for E. coli growth on the surface of the overlay. Only strains containing resistance to a specific antibiotic (or class) grow on this surface enabling rapid identification of the produced compound. This method has been successfully used for the identification of producers of known antibiotics and as a means to identify those producing novel compounds.
Topics: Anti-Bacterial Agents; Biological Products; Drug Discovery; Drug Resistance, Microbial; Escherichia coli; Mass Spectrometry
PubMed: 31680676
DOI: 10.3791/60536