Alterations in Membrane Phospholipid Fatty Acids of Gram-Positive Piezotolerant Bacterium Sporosarcina sp. DSK25 in Response to Growth Pressure

Pressure is an important thermodynamic property of the ocean and the deep biosphere that affects microbial physiology and biochemistry. Here, we report on our investigation of the response of Gram-positive piezotolerant bacterium Sporosarcina sp. DSK25 to hydrostatic pressure. Strain DSK25 responded in an adaptive manner to upshifts of growth pressure and showed systematic changes in phospholipid fatty acids. As the pressure increased from 0.1 to 10 MPa (Megapascal), unsaturated fatty acids in DSK25 increased from 21.7 to 31.1 % of total fatty acids, while the level of iso- and anteiso-branched fatty acids remained unchanged. At higher pressures (30, 50, and 60 MPa), the amount of unsaturated fatty acids decreased, and that of anteiso-branched fatty acids increased from 34.4 to 49.9 % at the expense of iso-branched fatty acids. For the first time, two polyunsaturated fatty acids (PUFA), 18:2n-6 and 18:2n-x, with the latter having much higher abundance than the former, were identified in DSK25. The concentration of the PUFA increased with growth pressure. These results indicate the involvement of unsaturated and methyl-branched fatty acids in the modulation of bacteria membrane fluidity and function over environmentally relevant parameter (pressure). Piezotolerant bacterium Sporosarcina sp. DSK25 appears to utilize two regulatory mechanisms for adaptation to high pressure, a rapid-responding mechanism on transient scale, expressed as increased biosynthesis of monounsaturated fatty acids, and a long-term adaptation mechanism in increased synthesis of anteiso-branched and polyunsaturated fatty acids. Our results further suggest that Gram-positive piezophilic bacteria respond differently than Gram-negative bacteria in adaptation to high pressure.     

THZ531 Induces a State of BRCAness in Multiple Myeloma Cells: Synthetic Lethality with Combination Treatment of THZ 531 with DNA Repair Inhibitors

Multiple myeloma (MM) is a hematological disease marked by abnormal growth of B cells in bone marrow. Inherent chromosomal instability and DNA damage are major hallmarks of MM, which implicates an aberrant DNA repair mechanism. Studies have implicated a role for CDK12 in the control of expression of DNA damage response genes. In this study, we examined the effect of a small molecule inhibitor of CDK12–THZ531 on MM cells. Treatment of MM cells with THZ531 led to heightened cell death accompanied by an extensive effect on gene expression changes. In particular, we observed downregulation of genes involved in DNA repair pathways. With this insight, we extended our study to identify synthetic lethal mechanisms that could be exploited for the treatment of MM cells. Combination of THZ531 with either DNA-PK inhibitor (KU-0060648) or PARP inhibitor (Olaparib) led to synergistic cell death. In addition, combination treatment of THZ531 with Olaparib significantly reduced tumor burden in animal models. Our findings suggest that using a CDK12 inhibitor in combination with other DNA repair inhibitors may establish an effective therapeutic regimen to benefit myeloma patients.     

Immunogenic Cell Death of Breast Cancer Stem Cells Induced by an Endoplasmic Reticulum-Targeting Copper(II) Complex

Immunogenic cell death (ICD) offers a method of stimulating the immune system to attack and remove cancer cells. We report a copper(II) complex containing a Schiff base ligand and a polypyridyl ligand, 4 , capable of inducing ICD in breast cancer stem cells (CSCs). Complex 4 kills both bulk breast cancer cells and breast CSCs at sub-micromolar concentrations. Notably, 4 exhibits greater potency (one order of magnitude) towards breast CSCs than salinomycin (an established breast CSC-potent agent) and cisplatin (a clinically approved anticancer drug). Epithelial spheroid studies show that 4 is able to selectively inhibit breast CSC-enriched HMLER-shEcad spheroid formation and viability over non-tumorigenic breast MCF10 A spheroids. Mechanistic studies show that 4 operates as a Type II ICD inducer. Specifically, 4 readily enters the endoplasmic reticulum (ER) of breast CSCs, elevates intracellular reactive oxygen species (ROS) levels, induces ER stress, evokes damage-associated molecular patterns (DAMPs), and promotes breast CSC phagocytosis by macrophages. As far as we are aware, 4 is the first metal complex to induce ICD in breast CSCs and promote their engulfment by immune cells.     

Investigation of structural,optical, electrical,and magnetic properties of Fe-doped La0.7Sr0.3MnO3 manganites

In this work, the physical properties of nanocrystalline samples of La_0.7Sr_0.3Mn_1−xFe_xO₃ (0.0 ≤ x ≤ 0.20) perovskite manganites synthesized by the reverse micelle (RM) technique were explored in detail. The phase purity, crystal structure, and crystallite size of the samples were determined using X-ray diffraction (XRD) and Fourier transform infrared (FTIR) spectroscopy. All the samples had rhombohedral crystal structure and crystallite size increased with increase in Fe content in La_0.7Sr_0.3MnO₃. The scanning electron micrographs (SEMs) exhibited smooth surface morphology and nonuniform shape of the particles. The optical properties studied using UV-visible absorption spectroscopy revealed a decrease in the absorbance and optical band gap with an increase in Fe content in La_0.7Sr_0.3MnO₃ compound. The temperature-dependent resistivity measurements revealed semiconducting nature of x = 0 and 0.1 samples up to the studied temperature range, while a metal-to-insulator transition was observed at higher Fe doping. Magnetic studies revealed weak ferromagnetism in all the samples and a reduction in the maximum magnetization with an increase in Fe content. A close correlation between electrical transport and magnetic properties was observed with the doping of Fe ion in La_0.7Sr_0.3MnO₃ at Mn site. These results advocate strong interactions associated with the double exchange mechanism among Fe³⁺ and Mn³⁺ ions.     

In vitro evaluation of Pediococcus acidilactici NCDC 252 for its probiotic attributes

In vitro probiotic attributes of Pediococcus acidilactici, viz. tolerance to acidity and bile salt, phenol resistance, lactic acid production, antioxidant activity, cell surface hydrophobicity, lysozyme resistance and the presence of β‐galactosidase, proteolytic, lipase and peptidoglycan hydrolase activities, were studied. The strain was found to be rich in different proteolytic activities, β‐galactosidase and antioxidant activities, and produced lactic acid. The studies conducted support that P. acidilactici national collection of dairy cultures (NCDC) 252 is a potential probiotic for humans with all the essential basic probiotic properties. This testing is useful to gain insight into this strain and its mechanism of action. All these attributes add to its therapeutic importance and industrial significance.     

Fabrication of Food Grade Vitamin E Nanoemulsion by Low Energy Approach,Characterization and Its Application

The present study was carried out to fabricate the food grade vitamin E acetate nanoemulsion using edible mustard oil and to evaluate its improved bioactivities. A food-grade vitamin E acetate nanoemulsion was fabricated using the edible mustard oil and surfactant Tween-80. Flocculation was not observed for 15 days. The nanoemulsion was characterized for droplet morphology and size distribution using atomic force microscope and zetasizer, respectively. We observe a stable nanoemulsion of spherical morphology and a size distribution of 86.45 ± 3.61 nm. Further, the high-performance liquid chromatography method was used to determine the vitamin E acetate concentration and encapsulation efficiency for the stable nanoemulsion. These nanoemulsions showed improved bioactivity, antioxidant, and antimicrobial activity and could be potentially used to increase the shelf life of fruit juice.     

Chemopreventive Effect of Different Ratios of Fish Oil and Corn Oil in Experimental Colon Carcinogenesis

n-3 Polyunsaturated fatty acids (PUFA) have a chemopreventive effect while n-6 PUFA promote carcinogenesis. The effect of these essential fatty acids may be related to oxidative stress. Therefore, the study was designed to evaluate the effect of different ratios of fish oil (FO) and corn oil (CO) in the prevention of colon cancer. Male Wistar rats were divided into control, dimethylhydrazine dihydrochloride (DMH) treated, FO + CO (1:1) and FO + CO (2.5:1). All the groups, except the control received a weekly injection of DMH for 4 weeks. The animals were sacrificed either 48 h later (initiation phase) or kept for 16 weeks (post initiation phase). DMH treatment in the initiation phase animals showed mild to moderate inflammation, decreased ROS and TrxR activity, increased antioxidants, apoptosis and ACF multiplicity. The post initiation study showed severe inflammation with hyperplasia, increased ACF multiplicity and ROS levels, a decrease in antioxidants and apoptosis. The FO + CO (1:1) treated animals showed severe inflammation, a decrease in ROS, an increase in antioxidants and apoptosis in the initiation phase. FO + CO (1:1) in the post initiation phase and FO + CO (2.5:1) in the initiation showed mild inflammation, increased ROS, apoptosis and decreased antioxidants. There was a decrease in ACF multiplicity and ROS levels, increased antioxidants and apoptosis in the post initiation phase study. The present study suggests that FO has a dose- and time-dependent chemopreventive effect in colon cancer mediated through oxidative stress and apoptosis.     

Weldability and microstructural variations in weldments of Ti-5Ta-1.8Nb alloy

The successful replacement of the present generation of corrosion-resistant materials (nitric acid-grade stainless steel and Ti) by Ti-5Ta-1.8Nb, which has better corrosion resistance, depends on its weldability characteristics. This article presents the results of a study on the fabrication, qualification, and microstructural characterization of the welds. Welding was carried out using the direct current electrode negative (DCEN) polarity tungsten inert gas (TIG) (manual) welding method with high-purity Ar shielding. Testing was carried out as per the ASME standard (section IX, welding and brazing). Qualification tests found that the weldment met the required properties. The weldment showed heterogeneous microstructures, which are rationalized based on differences in phase transformation mechanisms that are dictated by the thermal cycles experienced by various microscopic regions. The results, described in this article, confirm that the weldability of the developmental Ti-Ta-Nb alloy is excellent. A preliminary evaluation of the corrosion behavior of the welds showed rates comparable to that of the base metal, establishing that this alloy could be considered as an alternative material for use in highly corrosive environments.     

Kharitonov''s theorem revisited

A simple interpretation of Kharitonov's theorem is given.