292 research outputs found

    Immunopurification and characterization of a rape (Brassica napus L.) seedling lipase

    Get PDF
    Lipase or triacylglycerol acylhydrolase (E.C.3.1.1.3) was purified to homogeneity from rapeseed-germinated cotyledons (Brassica napus L.). The purification scheme involved homogenization, centrifugation, ultracentrifugation and affinity chromatography using polyclonal antibodies raised against porcine pancreatic lipase. The purified rapeseed lipase was homogenous and did not contain contaminating proteins detectable by SDS-PAGE and HPLC analysis. The specific activity of the purified preparation was increased about 1950 times, with an overall yield of 35%. The rapeseed lipase was found to be a cytosoluble, glycosylated and heat-labile serine-hydrolase. It was monomeric with a molecular mass of 38 kDa and a pH of 6.6. The purification method used in the present work is rapid, simple, and yields highly purified lipase. It may therefore be applicable in the purification of other uncharacterized plant lipases.Keywords: Brassica napus L., immuno-affinity, lipase, purification, triacylglycerol acyl hydrolaseAfrican Journal of Biotechnology Vol. 12(21), pp. 3224-323

    Use of natural fluorescent triacyglycerols from Parinari glaberrimum to detect low lipase activity in Brassica napus seedlings

    Get PDF
    Lipases (triacylglycerol acyl hydrolass EC 3.1.1.3) are defined as enzymes that catalyse the hydrolysis of triacylglycerols, releasing long-chain fatty acids. Germinating oilseeds have been explored as a possible source of lipases for biotechnological processing of oils and fats. However, purification and sensitive assays to detect a true lipase activity in plant cellular homogenates and culture media are required. The main aim of this study was to design a convenient, specific, sensitive and continuous lipase activity assay using natural long-chain triacylglycerols (TAGs). Oil was extracted from Parinari glaberrimum seed kernels and the purified TAGs were used as a substrate for detecting low levels of lipase activities. An increase in the fluorescence was observed, which is due to the parinaric acid released by various lipase activities. This increase in the fluorescence intensity is linear with time and proportional to the amount of lipase added. This new method, performed under non-oxidative conditions, was applied successfully to detect low lipase levels in crude protein extracts from plant seeds.Keywords: Brassica napus, fluorescent lipids, lipase assay, plant lipasesAfrican Journal of Biotechnology Vol. 12(23), pp. 3719-372

    BIBLIOGRAFI BERANOTASI SUBYEK PSIKOLOGI KEPRIBADIAN DAN PSIKOLOGI SOSIAL DI PERPUSTAKAAN FAKULTAS PSIKOLOGI UNAIR

    Get PDF
    ABSTRAK INI MEMBAHAS TENTANG : BIBLIOGRAFI BERANOTASI SUBYEK PSIKOLOGI KEPRIBADIAN DAN PSIKOLOGI SOSIAL DI PERPUSTAKAAN FAKULTAS PSIKOLOGI UNAI

    Prostaglandin D2 inhibits airway dendritic cell migration and function in steady state conditions by selective activation of the D prostanoid receptor 1

    Get PDF
    PGD(2) is the major mediator released by mast cells during allergic responses, and it acts through two different receptors, the D prostanoid receptor 1 (DP1) and DP2, also known as CRTH2. Recently, it has been shown that PGD(2) inhibits the migration of epidermal Langerhans cells to the skin draining lymph nodes (LNs) and affects the subsequent cutaneous inflammatory reaction. However, the role of PGD(2) in the pulmonary immune response remains unclear. Here, we show that the intratracheal instillation of FITC-OVA together with PGD(2) inhibits the migration of FITC(+) lung DC to draining LNs. This process is mimicked by the DP1 agonist BW245C, but not by the DP2 agonist DK-PGD(2). The ligation of DP1 inhibits the migration of FITC-OVA(+) DCs only temporarily, but still inhibits the proliferation of adoptively transferred, OVA-specific, CFSE-labeled, naive T cells in draining LNs. These T cells produced lower amounts of the T cell cytokines IL-4, IL-10, and IFN-gamma compared with T cells from mice that received FITC-OVA alone. Taken together, our data suggest that the activation of DP receptor by PGD(2) may represent a pathway to control airway DC migration and to limit the activation of T cells in the LNs under steady state conditions, possibly contributing to homeostasis in th

    Flat-gain wide-band erbium doped fiber amplifier by combining two difference doped fibers

    Get PDF
    A new erbium-doped fibre amplifier (EDFA) is demonstrated using a combination of newly developed Erbium Zirconia co-doped fiber (Zr-EDF) and the commercial silica-based Erbium-doped fiber (Si-EDF) as the gain medium. Both fibers have a very high concentration of erbium ion. A compact amplifier operating in C-band region is firstly reported using a double-pass configuration. It is shown that average gains of the proposed Zr-EDF amplifier are obtained at approximately 18 dB with a gain variation of ±2 dB within C-band region. A flat-gain and wide band operation is achieved by configuring the amplifier in two stages comprising a 2 m long Zr-EDF and 9 m long Si-EDF optimised for C- and L-band operations, respectively, in a double-pass parallel configuration. A chirp fibre Bragg grating (CFBG) is used in both stages to ensure double propagation of the signal and thus to increase the attainable gain in both C- and L-band regions. At an input signal power of 0 dBm, a flat gain of 15 dB is achieved with a gain variation of less than 0.5 dB within a wide wavelength range from 1530 to 1605 nm. The corresponding noise figure varies from 6.2 to 10.8 dB within this wavelength region

    Bilateral myositis ossificans of the masseter muscle after chemoradiotherapy and critical illness neuropathy- report of a rare entity and review of literature

    Get PDF
    Myositis ossificans in the head and neck is a rare heterotropic bone formation within a muscle. Besides fibrodysplasia ossificans progressiva, traumatic and neurogenic forms are described in the literature

    Alteration of renal respiratory Complex-III during experimental type-1 diabetes

    Get PDF
    <p>Abstract</p> <p>Background</p> <p>Diabetes has become the single most common cause for end-stage renal disease in the United States. It has been established that mitochondrial damage occurs during diabetes; however, little is known about what initiates mitochondrial injury and oxidant production during the early stages of diabetes. Inactivation of mitochondrial respiratory complexes or alteration of their critical subunits can lead to generation of mitochondrial oxidants, mitochondrial damage, and organ injury. Thus, one goal of this study was to determine the status of mitochondrial respiratory complexes in the rat kidney during the early stages of diabetes (5-weeks post streptozotocin injection).</p> <p>Methods</p> <p>Mitochondrial complex activity assays, blue native gel electrophoresis (BN-PAGE), Complex III immunoprecipitation, and an ATP assay were performed to examine the effects of diabetes on the status of respiratory complexes and energy levels in renal mitochondria. Creatinine clearance and urine albumin excretion were measured to assess the status of renal function in our model.</p> <p>Results</p> <p>Interestingly, of all four respiratory complexes only cytochrome c reductase (Complex-III) activity was significantly decreased, whereas two Complex III subunits, Core 2 protein and Rieske protein, were up regulated in the diabetic renal mitochondria. The BN-PAGE data suggested that Complex III failed to assemble correctly, which could also explain the compensatory upregulation of specific Complex III subunits. In addition, the renal F<sub>0</sub>F<sub>1</sub>-ATPase activity and ATP levels were increased during diabetes.</p> <p>Conclusion</p> <p>In summary, these findings show for the first time that early (and selective) inactivation of Complex-III may contribute to the mitochondrial oxidant production which occurs in the early stages of diabetes.</p

    Differentially Expressed RNA from Public Microarray Data Identifies Serum Protein Biomarkers for Cross-Organ Transplant Rejection and Other Conditions

    Get PDF
    Serum proteins are routinely used to diagnose diseases, but are hard to find due to low sensitivity in screening the serum proteome. Public repositories of microarray data, such as the Gene Expression Omnibus (GEO), contain RNA expression profiles for more than 16,000 biological conditions, covering more than 30% of United States mortality. We hypothesized that genes coding for serum- and urine-detectable proteins, and showing differential expression of RNA in disease-damaged tissues would make ideal diagnostic protein biomarkers for those diseases. We showed that predicted protein biomarkers are significantly enriched for known diagnostic protein biomarkers in 22 diseases, with enrichment significantly higher in diseases for which at least three datasets are available. We then used this strategy to search for new biomarkers indicating acute rejection (AR) across different types of transplanted solid organs. We integrated three biopsy-based microarray studies of AR from pediatric renal, adult renal and adult cardiac transplantation and identified 45 genes upregulated in all three. From this set, we chose 10 proteins for serum ELISA assays in 39 renal transplant patients, and discovered three that were significantly higher in AR. Interestingly, all three proteins were also significantly higher during AR in the 63 cardiac transplant recipients studied. Our best marker, serum PECAM1, identified renal AR with 89% sensitivity and 75% specificity, and also showed increased expression in AR by immunohistochemistry in renal, hepatic and cardiac transplant biopsies. Our results demonstrate that integrating gene expression microarray measurements from disease samples and even publicly-available data sets can be a powerful, fast, and cost-effective strategy for the discovery of new diagnostic serum protein biomarkers

    Flat-gain wide-band erbium doped fiber amplifier by combining two difference doped fibers

    Get PDF
    A new erbium-doped fibre amplifier (EDFA) is demonstrated using a combination of newly developed Erbium Zirconia co-doped fiber (Zr-EDF) and the commercial silica-based Erbium-doped fiber (Si-EDF) as the gain medium. Both fibers have a very high concentration of erbium ion. A compact amplifier operating in C-band region is firstly reported using a double-pass configuration. It is shown that average gains of the proposed Zr-EDF amplifier are obtained at approximately 18 dB with a gain variation of +-2 dB within C-band region. A flat-gain and wide band operation is achieved by configuring the amplifier in two stages comprising a 2 m long Zr-EDF and 9 m long Si-EDF optimised for C- and L-band operations, respectively, in a double-pass parallel configuration. A chirp fibre Bragg grating (CFBG) is used in both stages to ensure double propagation of the signal and thus to increase the attainable gain in both C- and L-band regions. At an input signal power of 0 dBm, a flat gain of 15 dB is achieved with a gain variation of less than 0.5 dB within a wide wavelength range from 1530 to 1605 nm. The corresponding noise figure varies from 6.2 to 10.8 dB within this wavelength regio
    corecore