Nucleic acid amplification technology, such as polymerase chain reaction (PCR), has enabled highly sensitive and specific disease detection and quantification, leading to more accurate diagnosis and treatment regimens. Lab-on-a-chip applications have developed methods to partition single biomolecules, such as DNA and RNA, into picoliter-sized droplets. These individual reaction vessels lead to digitization of PCR enabling improved time to detection and direct quantification of nucleic acids without a standard curve, therefore simplifying assay analysis. Though impactful, these improvements have generally been restricted to centralized laboratories with trained personnel and expensive equipment. To address these limitations and make this technology more applicable for a variety of settings, we have developed a statistical framework to apply to droplet PCR performed in polydisperse droplets prepared without any specialized equipment. The polydisperse droplet system allows for accurate quantification of droplet digital PCR (ddPCR) and reverse transcriptase droplet digital PCR (RT-ddPCR) that is comparable to commercially available systems such as BioRad's ddPCR. Additionally, this approach is compatible with a range of input sample volumes, extending the assay dynamic range beyond that of commercial ddPCR systems. In this work, we show that these ddPCR assays can reduce overall assay time while still providing quantitative results. We also report a multiplexed ddPCR assay and demonstrate proof-of-concept methods for rapid droplet preparation in multiple samples simultaneously. Our simple polydisperse droplet preparation and statistical framework can be extended to a variety of settings for the quantification of nucleic acids in complex samples.
Antibiotic resistance of bacteria stimulates the development of new treatment approaches. Piezoelectric-catalysis has attracted much attention due to the possibility to effectively provide antibacterial effect via generation of reactive oxygen species. However, the influence of the surface charge or potential of a piezopolymer on bacteria has not been sufficiently studied so far. This study reports the fabrication and characterization of thin films of piezoelectric polyhydroxybutyrate, polyvinylidene fluoride, and polyvinylidene fluoride trifluoroethylene as well as non-piezoelectric polycaprolactone polymers fabricated using solution casting approach. The piezoelectric coefficient (d33) and surface electric peak-to-peak potential generated by the cyclic mechanical stress applied to the films were measured. Neither any toxic effect of the polymer films nor ultrasound influence on Escherichia coli bacteria behavior is observed. However, significant inhibition of the growth of bacteria is revealed during mechanical stimulation of piezoelectric samples via ultrasound treatment. Thus, this study demonstrates clear bacteriostatic effect of piezoelectric polymers for different tissue engineering applications.
The antioxidant activity and immuno-tropic effects of lithium glutamate, lithium salicylate, lithium benzoate and lithium lactate have been investigated in this work, as a base for new psychotropic medicines. Methods: The antioxidant properties were studied by the voltammetric method. Phagocytic activity of neutrophilic leucocytes and the reaction of blastic transformation of lymphocytes were used as test for assessments of influence of the lithium compounds on the immune cells of human blood. Results: It was revealed absence of toxic action on human blood cells for all tested substances. Lithium benzoate showed the most significant stimulating influence on lymphocytes. Glutamate and benzoate lithium expressed scavenging activity vs oxygen radicals. Salicylate and benzoate lithium revealed significant phagocytosis stimulation effects. Conclusion: Investigated lithium salts expressed antioxidant activity and immunotropic effects, all investigated substance are of interest in medical application for mental diseases and comorbid pathology treatment.
The research of the degree of oil biodegradation by gas chromatography and individual classes of hydrocarbons by various strains of hydrocarbon-oxidizing microorganisms isolated from indigenous microflora of oil fields was carried out. It has been shown that some of the investigated strains of hydrocarbon-oxidizing microorganisms are 100% capable to biotransform naphthenes and olefins, showing high activity in the destruction of paraffins and isoparaffins. There are no signs of biodegradation of aromatic compounds due to the large duration of the process. All investigated strains of hydrocarbon-oxidizing microorganisms are largely able to reduce the total number of individual components of oil. The obtained data can be used to develop new biologics of the purpose
The selective activity of hydrocarbon-oxidizing microorganisms with regard to the degradation of alkanes, cycloalkanes, arenes was presented. The hydrocarbon-oxidizing activity of microorganisms of the genera Rhodococcus and Pseudomonas such as heptane, cyclohexane, toluene within the hydrocarbons destruction was determined. The growth rate for various hydrocarbons differs. Thus, the average specific growth rate of hydrocarbon-oxidizing microorganism (HOM) of the genus Rhodococcus is twice more than these substrates
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