oncogene-expression-analysis-with-alterations-ph-pancreatic-ductal

Oncogene Expression Analysis with Alterations in pH in a Pancreatic Ductal Cell Line

The fourth leading cause of cancer-related death, pancreatic ductal adenocarcinoma (PDAC), has a 12% five-year survival rate. This disease has a poor ...

Osmania University

The fungus Fusarium verticillioides MRC 826 (ascomycetes species), a toxigenic isolate is capable of synthesizing mycoferritin only upon induction with iron in yeast extract sucrose medium. The molecular mass, yield, iron and carbohydrate contents of the purified mycoferritin were 460 kDa, 0.010 mg/g of wet mycelia, 1.0 and 40.2%, respectively. Native gel electrophoresis of the mycoferritin revealed two bands possibly representing isoforms of ferritin. Subunit analysis by SDS-PAGE showed a single protein subunit of ~24 kDa suggesting similar sized subunits in the structure of apoferritin shell. Immunological cross reactivity was observed with the anti-fish liver ferritin. Transmission electron microscopy revealed an apparent particle size of 100 Å. N-terminal amino acid sequencing of mycoferritin showed identities with other eukaryotic ferritin sequences. The spectral characteristics were similar to equine spleen ferritin. However, circular dichroic spectra revealed a higher degree of helicity. Functionally, induction of mycoferritin minimizes the prooxidant role of iron.

Purification and characterization of mycoferritin from Fusarium verticillioides MRC 826.

The genotoxic potential of fumonisin B(1) (FB(1)) in vivo in BALB/c mice (male and female) was assessed by induction of micronuclei (MN) formation in bone marrow polychromatic erythrocytes. The ratio of polychromatic erythrocytes to normochromatic erythrocytes (PCE/NCE) was also determined. Mice were injected intraperitoneally (i.p.) with FB(1) at a low dose (0.1 mg kg(-1) body mass), middle dose (1.0 mg kg(-1) body mass) and high dose (10 mg kg(-1) body mass) as single and multiple doses in normal saline to test the genotoxicity. Mitomycin C, a known clastogen, was used as positive control. The frequency of MN and the PCE/NCE value in animals treated with FB(1) at low, middle, and high doses in single dose studies, and the frequency of MN in multiple dose studies, were statistically non-significant from that of the controls injected with saline only. The multiple dose studies at all doses revealed that the PCE/NCE value was found to be reduced upon exposure to FB(1) as compared to the controls. In animals injected with multiple low doses of FB(1), the PCE/NCE value was found to be 0.66 in males and 0.82 in the females; at multiple middle doses the value was 0.30 in males and 0.41 in the females and was statistically significant (p < 0.001); however, at multiple high doses, the ratio was found to be 0.36 in both males and females. The present study confirms that FB(1) is non-genotoxic in nature while the reduced ratio of PCE/NCE suggests the cytotoxic nature of FB(1).

Lack of micronuclei induction by fumonisin B(1) mycotoxin in BALB/c mice.

The effect of surfactants (two cationic, one anionic and three non-ionic) at 0.001, 0.01, 0.1 and 1.0 % concentrations on aflatoxin production, ergosterol content and sugar consumption by Aspergillus parasiticus (NRRL 2999) in YES liquid culture medium is reported. At 0.01% concentration, the cationic surfactants, cetyl dimethyl ammonium bromide (CDAB) and dodecyl trimethyl ammonium bromide (DTAB), and the anionic surfactant, sodium dodecyl sulfate (SDS), completely inhibited spore germination, while DTAB also inhibited the production of ergosterol and toxin (p < 0.05). At a concentration of 0.001%, CDAB was found to enhance toxin production, while SDS was found to inhibit it when compared with other surfactants. Non-ionic surfactants, polyoxyethylene sorbitan monolaurate (Tween-20), polyoxyethylene lauryl ether (Brij-35) and ethoxylated p-tert-octylphenol (Triton X-100) delayed the spore germination up to day 5 at all concentrations and inhibited toxin and ergosterol production at 0.001% concentration. The affect was found to be dose-dependent from 0.001% to 1%, for Triton X-100 only. Positive correlation between ergosterol content and toxin production in the presence of different surfactants at various time periods (3, 5, 7, 9 and 12 days) was found. Tween-20 was most effective in inhibiting toxin production on day 7, when aflatoxin production was found to be maximal in control group. Sugar consumption was directly proportional to the ergosterol content, showing a significant correlation with aflatoxin production.

Effect of various surfactants (cationic, anionic and non-ionic) on the growth of Aspergillus parasiticus (NRRL 2999) in relation to aflatoxin production.

Human serum albumin acts as a carrier protein to a variety of drugs and aids their transport. Andrographis paniculata, a herbal plant has been used as a source of traditional medicine in the Asian countries. Among the various constituents of this plant, andrographolide is the most active and is being used from centuries in the treatment of many chronic and infectious diseases.

Binding Studies of Andrographolide with Human Serum Albumin: Molecular Docking, Chromatographic and Spectroscopic Studies.

Bacillus thuringiensis (Bt) synthesizes Cry1Ac protein, which is toxic to many lepidopteran pests, and the cry1ac gene has been expressed in several transgenic crop plants. The Cry1Ac protein has been isolated from Bt kurstaki HD73 and purified to homogeneity. Polyclonal antibodies were raised against purified Cry1Ac in rabbits and goat. Sandwich ELISA was developed for Cry1Ac using goat IgG as a coating antibody, and affinity-purified rabbit IgG as the primary antibody.

Immuno-analytical method development for detection of transgenic Cry1Ac protein and its validation.

Green synthesis of organic Pt-nanocomposite was accomplished using carboplatin as a precursor and novel biopolymer - gum kondagogu (GK) as a reducing agent. The synthesised GK stabilised organic Pt-nanocomposite (GKCPt NC) was characterised by different analytical techniques such as ultraviolet-visible spectroscopy, nanoparticle analyser, scanning electron microscopy and energy dispersive X-ray analysis, X-ray diffraction (XRD), Fourier-transform infrared spectroscopy, transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS) and inductively coupled plasma optical emission spectrophotometer. The XRD pattern established the amorphous nature of GKCPt NC. TEM analysis revealed the homogeneous, monodisperse and spherical nature, with Pt metal size of 3.08 ± 0.62 nm. The binding energy at 71.2 and 74.6 eV show the presence of metallic platinum, Pt(0) confirmed by XPS studies. Further, in vitro radical scavenging and antitumour activity of GKCPt NC have been investigated. In comparison to GK and carboplatin, GKCPt NC showed superior 1, 1-diphenyl-2-picrylhydrazyle activity of 87.82%, whereas 2, 2-azinobis-(3-ethylbenzthinzoline-6-sulphonic acid) activity was 38.50%, respectively. In vitro studies of the antitumour property of GK, GKCPt NC and carboplatin were evaluated by potato disc tumour bioassay model. The efficacy of synthesised GKCPt NC concentration (IC) on tumour inhibition was found to be 2.04-fold lower as compared to carboplatin. Overall, the synthesised GKCPt NC shows both antitumour and antioxidant properties when compared to the original drug - carboplatin and might have promising applications in cancer therapy.

Synthesis and characterisation of novel biopolymer stabilised organic Pt-nanocomposite: assessment of its antioxidant and antitumour properties.

Bimetallic nanoparticles (BNPs) constitute two different metal elements and exhibit relatively superior mechanistic and catalytic efficacies owing to their synergistic functions over monometallic nanoparticles. In the present study various bimetallic Ag-Au, Ag-Pd, Au-Pd nanoparticles were synthesized using a natural biopolymer gum kondagogu (GK) as a reducing and capping agent, by a simple and cost-effective method. The synthesized BNPs when characterized using UV-vis spectroscopy revealed a specific surface plasmon resonance band (SPR) of each nanocomposite. The average particle size of Ag-Au, Ag-Pd, and Au-Pd BNPs was found to be 23 ± 10.3, 21 ± 7.6, and 23 ± 9.4 nm respectively based on transmission electron microscopy analysis. Surface morphology and functional groups on the gum matrix of GK-BNPs were analyzed by XRD and FT-IR respectively. The bimetallic nanocomposites were evaluated for their catalytic reduction of 4-nitrophenol (4-NP) to 4-aminophenol in the presence of NaBH. The kinetic studies performed, depicted rate constants for Ag-Au, Ag-Pd, and Au-PdNPs as 0.31, 0.39, and 0.28 min respectively. The catalytic efficiencies of three bimetallic nanocomposites were of the following order Ag-Pd > Ag-Au > Au-Pd. This study establishes the catalytic potentials of the three different bimetallic nanocomposites in the reduction of 4-NP an environmental pollutant, and the impact of their synergistic property.

Bimetallic nanocomposite (Ag-Au, Ag-Pd, Au-Pd) synthesis using gum kondagogu a natural biopolymer and their catalytic potentials in the degradation of 4-nitrophenol.

The progression of chronic pancreatitis (CP), an inflammatory disease of the pancreas, causes pancreatic stones to form within the pancreatic ductal lumen/parenchyma, which occurs via protein plug formation. Pain is the most common symptom that necessitates clinical attention, and pain relief is the therapeutic goal for these patients. Endoscopic therapy and surgery are complimentary forms of therapy for pain relief. This study was envisaged to clarify the mechanism by which protein plug/soft stones form in pancreatic ducts prior to undergoing calcification.

Alterations in the pH of pancreatic juice are associated with chymotrypsin C inactivation and lithostathine precipitation in chronic pancreatitis patients: a proteomic approach.

Silver nanoparticles (AgNPs) have been widely utilized in various biomedical and antimicrobial technologies, displaying broad-spectrum activities against Gram-negative and Gram-positive bacteria including multidrug-resistant strains. However, the emergence of resistance to AgNPs upon repeated exposure and the survival of bacteria after initial exposure to antimicrobial agents pose a threat, as they may lead to the development of new resistant populations. To combat the early stages of antibacterial resistance, systematic analysis is essential to understand the immediate response of bacteria to antimicrobial agents. In this study, green-synthesized AgNPs with a diameter of approximately 14 nm were exposed toat three different inhibitory concentrations and at two different time intervals (1 and 4 h) to investigate the perturbations in the metabolome using liquid chromatography-high-resolution mass spectrometry. MetaboAnalyst 5.0 was employed for univariate and multivariate analysis, and the affected metabolic pathways were constructed using a variable important in projection scores above 1 from PLS-DA. The study revealed significant alterations in metabolites associated with cell wall synthesis, energy metabolism, nucleotide metabolism, the TCA cycle, and anaplerotic intermediates of the TCA cycle. Our investigation aimed to comprehensively understand the effects of green-synthesized AgNPs on metabolism, providing a more precise snapshot of the bacterium's physiological state through metabolomics approach.

Karuna Rupula

Department of Biochemistry

Osmania University

Oncogene Expression Analysis with Alterations in pH in a Pancreatic Ductal Cell Line

Karuna Rupula

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Biochemistry

Osmania University

Oncogene Expression Analysis with Alterations in pH in a Pancreatic Ductal Cell Line

Department of Biochemistry