Induction of HRR genes and inhibition of DNMT1 is associated with anthracycline anti-tumor antibiotic-tolerant breast carcinoma cells

Molecular and Cellular Biochemistry - The aim of the study was to understand the role of homologous recombination repair (HRR) pathway genes in development of chemotolerance in breast cancer (BC)....     

Production of Tag-Free Recombinant Fusion Protein Encompassing Promiscuous T Cell Epitope of Tetanus Toxoid and Dog Zona Pellucida Glycoprotein-3 for Contraceptive Vaccine Development

Affinity tags can interfere in various physicochemical properties and immunogenicity of the recombinant proteins. In the present study, tag-free recombinant fusion protein encompassing promiscuous T cell epitope of tetanus toxoid [TT; amino acid (aa) residues 830-844] followed by dilysine linker and dog zona pellucida glycoprotein-3 (ZP3; aa residues 23-348) (TT–KK–ZP3) was expressed in Escherichia coli. The recombinant protein, expressed as inclusion bodies (IBs), was purified by isolation of IBs, processed to remove host cell proteins, followed by solubilization and refolding. A specific 39 kDa protein including ZP3 was identified by SDS-PAGE. CD spectra showed the presence of α-helices and β-sheets, and fluorescent spectroscopy revealed emission maxima of 265 A.U. at 339 nm for refolded protein and showed red shift in the presence of 6 M guanidine hydrochloride. Immunization of inbred FvB/J female mice with purified recombinant TT–KK–ZP3 (25 μg/animal) led to generation of high antibody titers against the recombinant protein. The antibodies reacted specifically with ZP matrix surrounding mouse oocytes. Immunized mice showed significant reduction in fertility as compared to the control group. The studies described herein provide a simple method to produce and purify tag-free recombinant protein for the development of a contraceptive vaccine.     

Allantoin mediated regulation of miRNAs for short term salinity stress tolerance in Oryza sativa L. cv. IR-29

Journal of Plant Biochemistry and Biotechnology - Salinity affects plant in a variety of ways, including ion toxicity, osmotic stress and oxidative damage. Allantoin is a nitrogenous compound...     

In vitro regeneration of Semecarpus anacardium L. from axenic seedling-derived nodal explants

Semecarpus anacardium (Anacardiaceae), a deciduous forest tree, is a potent source of medicinal compounds. Poor seed viability of this species limits the conventional propagation practice. Proliferation of shoots from axillary meristem was achieved in semisolid WPM medium supplemented with BAP 4.44 μM and KN 4.64 μM. Factors including culture vessels, gelling agents and antioxidants were identified and optimized for proliferation and growth of shoots in vitro. Cotton-plugged culture vessels were more favorable. Phytagel 0.2% as gelling agent and activated charcoal 0.2% as antioxidant were superior to other agents and antioxidants tested. All the shoots rooted in half-strength WPM liquid medium with IBA 2.46 μM. Rooted shoots survived (91%) in the soil–sand 1:1 mixture. Ex vitro rooting of shoots and hardening of plants were achieved in 80% of the explants in the soil–sand mixture. Hardened plants were maintained in a greenhouse. This is the first report on in vitro regeneration of Semecarpus anacardium.     

Functional analysis of RXLR effectors from the New Zealand kauri dieback pathogen Phytophthora agathidicida

New Zealand kauri is an ancient, iconic, gymnosperm tree species that is under threat from a lethal dieback disease caused by the oomycete Phytophthora agathidicida. To gain insight into this pathogen, we determined whether proteinaceous effectors of P. agathidicida interact with the immune system of a model angiosperm, Nicotiana, as previously shown for Phytophthora pathogens of angiosperms. From the P. agathidicida genome, we defined and analysed a set of RXLR effectors, a class of proteins that typically have important roles in suppressing or activating the plant immune system. RXLRs were screened for their ability to activate or suppress the Nicotiana plant immune system using Agrobacterium tumefaciens transient transformation assays. Nine P. agathidicida RXLRs triggered cell death or suppressed plant immunity in Nicotiana, of which three were expressed in kauri. For the most highly expressed, P. agathidicida (Pa) RXLR24, candidate cognate immune receptors associated with cell death were identified in Nicotiana benthamiana using RNA silencing-based approaches. Our results show that RXLRs of a pathogen of gymnosperms can interact with the immune system of an angiosperm species. This study provides an important foundation for studying the molecular basis of plant–pathogen interactions in gymnosperm forest trees, including kauri.     

Comparison of the elution characteristics of individual forms of lovastatin in both isocratic and gradient modes and HPLC-PDA method development for pure and fermentation-derived lovastatin

The elution characteristics of lovastatin were studied by varying the composition of mobile phase in both isocratic and gradient elution modes to comprehend the role of organic modifier and acidifier on the overall analysis time and retention time of individual forms of lovastatin. Acetonitrile has influenced on the overall analysis time, whereas the acidifier determines the retention time of hydroxy acid form of lovastatin and the retention time gap between the individual forms. A combination of acetonitrile and 0.1% trifluoroacetic acid (TFA) (60:40, v/v) in isocratic elution mode eluted both hydroxy acid and lactone forms of lovastatin at 4.5 and 5.4?min, respectively. This appears to be a better approach for the separation of pharmaceutical and clinical lovastatin samples. At isocratic elution mode, a mixture of acetonitrile and either 0.05% TFA or 0.1% H₃PO₄ of 60:40 (v/v) has eluted both hydroxy acid and lactone forms of lovastatin at 10?±?0.5 and 17?±?0.5?min, respectively. This is suitable for the fermentation-derived samples or for the complex mixtures of structural analogs. The fermentation broth (pH not adjusted) extracted with ethyl acetate at a ratio of 1:1 (v/v) at 60°C for 30?min was the optimal extraction condition for lovastatin.     

Comparative analysis of calcium gluconate and sodium gluconate techniques for the production of gluconic acid by Aspergillus niger

Sodium gluconate and calcium gluconate methods are important techniques available for gluconic acid fermentation. The comparative analysis of these fermentations has been addressed using Aspergillus niger. The techniques are equally influenced by the spores age in slant growth, inoculum level in germination and production media, different levels of Fe, Cu, Zn and Mn. Sodium gluconate method is promising with respect to lesser time for slant age (3 d) and lesser time of fermentation (6 d) compared to the calcium gluconate method (slant age — 6 d, and time of fermentation — 7 d).