Biomedical subjects

Mulpuri V Rao

Publications and source records attributed to Mulpuri V Rao.

4 recordsLinked to original sources

PubMed2006

Capillarity induced solvent-actuated bonding of polymeric microfluidic devices.

Rapid, robust, and economical fabrication of fluidic microchannels is of fundamental importance for the successful development of disposable lab-on-a-chip devices. In this work, we present a solvent-actuated bonding method for fabricating polymeric microfluidic devices at room temperature. A PMMA sheet with an imprinted microchannel was clamped to a blank PMMA sheet, and then 80 +/- 5 muL of acetone (bonding solvent) was introduced at one end of the fluidic channel and aspirated out at the other end. As the solvent moved down the channel, capillary forces drew a fraction of the solvent into the interstitial space between the two polymeric substrates. After aspiration, the assembly was incubated in the clamp for 5 min for effective bond formation. The quantity of the bonding solvent, its water content and flow rate, along with residence time in the channel were found to have significant impact on the bond quality and the channel integrity. Microfluidic electrophoretic separations of a 400-base DNA ladder were performed in devices fabricated using this method in less than 8 min with efficiencies routinely between 2 x 10(6) and 3 x 10(6) plates/m. The simplicity and economy of this technique make it amenable for automation and mass production, which could make polymeric substrates more attractive for single-use chemical analysis devices.

Journal Article↗

PubMed2006

Surface modification of poly(methyl methacrylate) for improved adsorption of wall coating polymers for microchip electrophoresis.

The development of rapid and simple wall coating strategies for high-efficiency electrophoretic separation of DNA is of crucial importance for the successful implementation of miniaturized polymeric DNA analysis systems. In this report, we characterize and compare different methods for the chemical modification of poly(methyl methacrylate) (PMMA) surfaces for the application of wall coating polymers. PMMA surfaces coated with 40 mol% diethylacrylamide and 60 mol% dimethylacrylamide are compared to the PMMA surfaces first oxidized and then coated with hydroxypropylmethyl-cellulose or poly(vinyl alcohol) (PVA). PMMA oxidation was accomplished with UV/ozone or an aqueous solution of HNO(3) to yield hydrogen-bond donors for the spontaneous adsorption of the coating polymers. Contact angle measurements of UV/ozone exposed PMMA surfaces indicate increase in hydrophilicity, and polymer coated surfaces show a strong dependence on the coating polymer and the oxidation method. Fast and repeatable electrophoretic separations of a 10-base and 20-base DNA ladder were performed in PMMA micro CE devices. All analyses were completed in less than 10 min, resulting in the number of theoretical plates as high as 583 000 in a 7.7 cm long separation channel. The duration of UV/ozone treatment was found to have a considerable impact on separation performance. The microchips irradiated with UV for 10 min and coated with PVA as well as the microchips treated with HNO(3) and coated with HPMC were found to have the best separation performance. These results demonstrate facile and robust methods for the surface modification of PMMA enabling low-cost single use devices for electrophoretic DNA separations.

Acrylamides↗

PubMed2003

The auxin-induced transcriptome for etiolated Arabidopsis seedlings using a structure/function approach.

To increase our understanding of the mode of action of auxin, we analyzed auxin-induced changes in the Arabidopsis transcriptome with microarrays representing 20426 Arabidopsis genes. Treatment of etiolated seedlings with low concentrations of the auxin, indole-3-acetic acid (IAA), decreased the expression levels of 23 genes, whereas it increased the expression levels of 47 genes within 20 min. After 40 min, the directional trend in genomic change was predominantly an increase in gene expression. Among the most rapidly induced changes are those in genes encoding transcription factors. Promoter regions of transiently induced genes contained DNA motifs that bind auxin response (ARFAT) and silence element binding factors whereas genes induced by IAA during the entire experimental period contained MYC and ARFAT DNA motifs at higher frequencies. Six structurally diverse auxins were analyzed to determine genes that are unique to a specific auxin, as well as a common set of genes that are rapidly regulated by all tested auxins, thus enabling the identification of shared DNA motifs. In addition to ARFAT, analysis of promoter regions of genes induced by all six auxins revealed the presence of an abscisic-acid-responsive DC3 promoter-binding factor and low temperature responsive elements suggesting a possible role for abscisic acid in modulating auxin-induced responses.

5' Untranslated Regions↗

PubMed2002

Ozone-induced ethylene production is dependent on salicylic acid, and both salicylic acid and ethylene act in concert to regulate ozone-induced cell death.

Ethylene is known to influence plant defense responses including cell death in response to both biotic and abiotic stress factors. However, whether ethylene acts alone or in conjunction with other signaling pathways is not clearly understood. Ethylene overproducer mutants, eto1 and eto3, produced high levels of ethylene and developed necrotic lesions in response to an acute O3 exposure that does not induce lesions in O3-tolerant wild-type Col-0 plants. Treatment of plants with ethylene inhibitors completely blocked O3-induced ethylene production and partially attenuated O3-induced cell death. Analyses of the responses of molecular markers of specific signaling pathways indicated a relationship between salicylic acid (SA)- and ethylene-signaling pathways and O3 sensitivity. Both eto1 and eto3 plants constitutively accumulated threefold higher levels of total SA and exhibited a rapid increase in free SA and ethylene levels prior to lesion formation in response to O3 exposure. SA pre-treatments increased O3 sensitivity of Col-0, suggesting that constitutive high SA levels prime leaf tissue to exhibit increased magnitude of O3-induced cell death. NahG and npr1 plants compromised in SA signaling failed to produce ethylene in response to O3 and other stress factors suggesting that SA is required for stress-induced ethylene production. Furthermore, NahG expression in the dominant eto3 mutant attenuated ethylene-dependent PR4 expression and rescued the O3-induced HR (hypersensitive response) cell death phenotype exhibited by eto3 plants. Our results suggest that both SA and ethylene act in concert to influence cell death in O3-sensitive genotypes, and that O3-induced ethylene production is dependent on SA.

Arabidopsis↗