Titanium dioxide nanoparticles preferentially bind in subdomains IB,IIA of HSA and minor groove of DNA

Titanium dioxide nanoparticles (TiO₂-NPs) interaction with human serum albumin (HSA) and DNA was studied by UV–visible spectroscopy, spectrofluorescence, circular dichroism (CD), and transmission electron microscopy (TEM) to analyze the binding parameters and protein corona formation. TEM revealed protein corona formation on TiO₂-NPs surface due to adsorption of HSA. Intrinsic fluorescence quenching data suggested significant binding of TiO₂-NPs (avg. size 14.0 nm) with HSA. The Stern–Volmer constant (K_sv) was determined to be 7.6 × 10² M^?1 (r² = 0.98), whereas the binding constant (K_a) and number of binding sites (n) were assessed to be 5.82 × 10² M^?1 and 0.97, respectively. Synchronous fluorescence revealed an apparent decrease in fluorescence intensity with a red shift of 2 nm at Δλ = 15 nm and Δλ = 60 nm. UV–visible analysis also provided the binding constant values for TiO₂-NPs–HSA and TiO₂-NPs-DNA complexes as 2.8 × 10² M^?1 and 5.4 × 10³ M^?1. The CD data demonstrated loss in α-helicity of HSA and transformation into β-sheet, suggesting structural alterations by TiO₂-NPs. The docking analysis of TiO₂-NPs with HSA revealed its preferential binding with aromatic and non-aromatic amino acids in subdomain IIA and IB hydrophobic cavity of HSA. Also, the TiO₂-NPs docking revealed the selective binding with A-T bases in minor groove of DNA.     

Comparison of development and demographic parameters of Diuraphis noxia (Hem., Aphididae) and its parasitoid,Diaeretiella rapae (Hym., Braconidae: Aphidiinae)

Demographic parameters of Diuraphis noxia (Mordvilko) and its parasitoid, Diaeretiella rapae (McIntosh) were investigated under laboratory conditions at temperatures of 20 ± 1°C, 60 ± 5% relative humidity of 60 ± 5% and a photoperiod of 16: 8 (light: dark) hours. The survival rates (l_x ) and the life expectancy (e_x ) at the beginning of adult emergence were 86% and 25.81 days for D. noxia and 100% and 7.36 days for D. rapae, respectively. Based on the Weibull distribution parameters, the survival curves were type I for both D. noxia and D. rapae, which indicates that mortality mostly occurred in older stages. The r _m-values in aphid and parasitoid were obtained as 0.22 ± 0.002 and 0.19 ± 0.003 day^?1, respectively. Mean generation time (T) and doubling time (DT) were 15.93 ± 0.202 and 3.15 ± 0.031 days for the aphid and 15.33 ± 0.071 and 3.67 ± 0.054 days for the parasitoid, respectively. The R₀ -values of D. noxia and D. rapae were evaluated 33.19 ± 0.961 and 18.07 ± 0.761 females/female/generation, respectively. The gross and net fecundity rates were 59.21 ± 1.94 and 32.825 ± 0.972 nymphs/female/day for the aphid and 37.59 ± 1.46, and 33.8 ± 1.51 eggs/female/day for the parasitoid, respectively. The results of this research indicated that D. rapae is an adequate parasitoid for control of Russian wheat aphid.     

Hypoxia impairs visual acuity in snapper (Pagrus auratus)

We investigated the effect of environmental hypoxia on vision in snapper (Pagrus auratus). Juvenile snapper inhabit estuarine environments where oxygen conditions fluctuate on a seasonal basis. Optomotor experiments demonstrated that visual acuity is impaired by environmental hypoxia, but not until levels approach the critical oxygen tension (P _crit) of this species (around 25 % air-saturated seawater). In 100, 80, and 60 % air-saturated seawater, a positive optomotor response was present at a minimum separable angle (M _SA) of 1°. In 40 % air-saturated seawater, vision was partially impaired with positive responses at M _SAs of 2° and above. However, in 25 % air-saturated seawater, visual acuity was seriously impaired, with positive responses only present at M _SAs of 6° and above. Snapper were found to possess a choroid rete, facilitating the maintenance of high ocular oxygen partial pressures (PO₂) during normoxia and moderate hypoxia (PO₂, between 269 and 290 mmHg). However, at 40 and 25 % water oxygen saturation, ocular PO₂ was reduced to below 175 mmHg, which is perhaps linked to impairment of visual acuity in these conditions. The ability to preserve visual function during moderate hypoxia is beneficial for the maintenance of a visual lifestyle in the fluctuating oxygen environments of estuaries.     

Toxic effect of deltamethrin on the life table and development of Helicoverpa armigera (Hubner)

Effect of deltamethrin (Decis 2.8% EC) on the fitness of Helicoverpa armigera (Hübner) was determined using demographic studies. Survivorship (I_x) and expectancy (e_x) was high with commencement of age and gradually decreased with progression of age in the exposed and unexposed groups. Highest number of unhatched eggs (~20%) was recorded with LC₅₀ (79.0 mg/L) opposed to 1% in the control. Daily fecundity rate was significantly greater (200 females/day) in the control than 126 in LC₂₀ (5.13 mg/L). Potential fecundity was lowest (179 females/female/generation) with LC₅₀ treatment than 675 in the control. Smallest value of intrinsic rate of increase (r_m) occurred at LC₅₀ (0.0393 females/female/day) while the highest value (0.0539) was at control. Overall larval development was significantly reduced to 18.0 days with LC₅₀ opposed to 24.1 days at control. Pupal period was shortened to 7.6 days with LC₅₀, while it recorded 9.3 days in control.     

New cytological data in Ophrys sect. Pseudophrys Godfery and comparative karyomorphological studies in Ophrys L. (Orchidaceae)

Eight species of Ophrys sect. Pseudophrys (Orchidaceae) were cytogenetically studied. The analysed species possess the most symmetrical karyotypes of the genus (M_CA ranged from 10.21 to 15.87 and CV_CL from 19.61 to 23.93) with 2n = 2x = 36, being composed of mainly metacentric chromosomes. The karyotype formulae were: 36m for Ophrys archimedea, 32m + 4sm for O. flammeola, 32m + 4sm for O. funerea, 36m for O. laurensis, 36m for O. lojaconoi, 34m + 2sm for O. lucifera, 34m + 2sm for O. obaesa and 36m for O. pallida. Karyotype morphometric characters were evaluated by calculating M_CA and CV_CL for the assessment of karyotype asymmetry, and CV_CI for the evaluation of heterogeneity in the position of the centromeres. The relationships between species were thus finally elucidated. The species characterised by wide distribution show greater karyomorphological distance than those with restricted distribution. The possible evolutionary role of chromosomal rearrangements as well as gene mutations in the speciation of Ophrys is discussed.     

Design,synthesis, MTT assay,DNA interaction studies of platinum(II) complexes

The square planar Pt(II) complexes of the type [Pt(Lⁿ)(Cl₂)] (where Lⁿ = L^1?3 = thiophene-2-carboxamide derivatives and L^4?6 = thiophene-2-carbothioamide derivatives) have been synthesized and characterized by physicochemical and various spectroscopic studies. MIC method was employed to inference the antibacterial potency of complexes in reference to free ligands and metal salt. Characteristic binding constant (K_b) and binding mode of complexes with calf thymus DNA (CT-DNA) were determined using absorption titration (0.76–1.61 × 10⁵ M^?1), hydrodynamic chain length assay and fluorescence quenching analysis, deducing the partial intercalative mode of binding. Molecular docking calculation displayed free energy of binding in the range of –260.06 to –219.63 kJmol^?1. The nuclease profile of complexes towards pUC19 DNA shows that the complexes cleave DNA more efficiently compared to their respective metal salt. Cytotoxicity profile of the complexes on the brine shrimp shows that all the complex exhibit noteworthy cytotoxic activity with LC₅₀ values ranging from 7.87 to 15.94 μg/mL. The complexes have been evaluated for cell proliferation potential in human colon carcinoma cells (HCT 116) and IC₅₀ value of complexes by MTT assay (IC₅₀ = 125–1000 μg/mL).     

Synthesis and biological evaluation of a new dysprosium(III) complex containing 2,9-dimethyl 1,10-phenanthroline

The binding of [Dy(dmp)₂Cl₃(OH₂)], where dmp is 2,9-dimethyl 1,10-phenanthroline, with Fish salmon DNA (FS-DNA) is investigated by absorption and emission spectroscopy, quenching studies, salt dependent, and gel electrophoresis. The binding constant (K_b) of the interaction is calculated as (1.27 ± .05) × 10⁵ M^?1 from absorption spectral titration data. The Stern–Volmer constant (K_SV), thermodynamic parameters involves ΔG°, ?H°, and ?S° are calculated by fluorescent data and Van’t Hoff equation. The thermodynamic studies show that the reaction for the binding of the complex with FS-DNA is endothermic and entropically driven (ΔS° > 0, ΔH° > 0). The effect of the complex concentration on FS-DNA cleavage reactions is also investigated by gel electrophoresis. Furthermore, the Dy(III) complex has been screened for its antibacterial activity. The experimental results suggest that the Dy(III) complex binds significantly to FS-DNA by hydrophobic groove binding mode and the complex has more efficient antibacterial activity compared to its metal salt.     

Biology and fertility life table of the greenbug,Schizaphis graminum (Hemiptera: Aphididae) on the resistant winter wheat cultivar (Pishgam) in Iran

The greenbug, Schizaphis graminum (Rondani) (Hemiptera: Aphididae) has been known as a major pest of small grains, particularly wheat, worldwide. In this study, the effect of new wheat cultivar (Pishgam) for cold regions on biological characteristics of greenbug was investigated in a greenhouse at 25 ± 2 °C, 55 ± 10% relative humidity and a photoperiod of 16:8 h (L:D). The raw data were analysed based on the age-stage, two-sex life table. The intrinsic rate of increase (r), the finite rate of increase (λ), the net reproduction rate (R₀) and the mean generation time (T) of greenbug were 0.313 ± 0.0019, 1.36 ± 0.0027 females/female/day, 83.33 ± 0.331 females/female and 14.11 ± 0.09 days, respectively. The life expectancy of a nymph is 43.57 days. The maximum reproductive value of females is on the 16th day which coincides with the total pre-reproduction period counted from birth. Hence, the present results may provide helpful information for comprehensive IPM programme of greenbug on this variety in cold regions of Iran. Result revealed that nymphal survival rate of the aphid was 100% on studied cultivar like that on sensitive host plant cultivar.     

Prominence of three diets on life table parameters for Chrysoperla carnea (Neuroptera: Chrysopidae) to mass rearing under laboratory conditions

Chrysoperla carnea (Steph.) is a general predator reared in industrial scale. Different eggs of moth were used to rear C. carnea, but stabled moth colony needed expensive equipment and is costly. In this research, we surveyed appropriate diet to mass rearing. For this purpose, 100 same old (24H) eggs of C. carnea, separately, were selected randomly from the mass culture of C. carnea which was reared on the egg of flour moth Anagasta kuehniella (Zeller), artificial diet and semi-artificial diet under laboratory conditions (25 ± 5°C, 65 ± 5% RH and L–D: 16–8). These results showed reduction process e_x (expectation of life table at age X) and the survival curve was convex (K-Strategy). Also L_x in appearing of adults that fed on egg of flour moth, artificial diet and semi-artificial diet were 0.76, 0.4 and 0.9 which implied that 24, 60 and 10% of cohort were dead before reaching adult stage. Eggs produced by each female were recorded daily until all females died. The parameters were estimated using Carey’s (1993) method. Gross (GRR) and net (R₀) reproductive rates of predator on A. kuehniella, artificial diet and semi artificial diets were 225.5 ± 3.45, 72.4 ± 3.5, 267.8 ± 4.8 and 180.12 ± 2.3, 24.33 ± 4.3, 254.05+3.3 (female/female/generation), respectively. Mean generation time (T), Doubling time (DT), Intrinsic rate of increase (r_m ) and Finite rate of increase (λ) of predator on A. kuehniella, artificial diet and semi artificial diet were 31.9 ± 0.71, 42.87 ± 0.45, 29.79 ± 0.57 (days); 4.27 ± 0.03, 9.36 ± 0.06, 3.74 ± 0.05 (days); 0.162 ± 0.001, 0.074 ± 0.003, 1.185 ± 0.002; and 1.175 ± 0.001, 1.076 ± 0.002, 1.203 ± 0.002 (female/female/day), respectively. This research indicated that semi-artificial diet is a suitable prey for the predator.     

Peptidic models for the binding of Pb(II), Bi(III) and Cd(II) to mononuclear thiolate binding sites

Herein, we evaluate the binding of Pb(II) and Bi(III) to cysteine-substituted versions of the TRI peptides [AcG-(LKALEEK)₄G-NH₂] which have previously been shown to bind Hg(II) and Cd(II) in unusual geometries as compared with small-molecule thiol ligands in aqueous solutions. Studies of Pb(II) and Bi(III) with the peptides give rise to complexes consistent with the metal ions bound to three sulfur atoms with M–S distances of 2.63 and 2.54 Å, respectively. Competition experiments between the metal ions Pb(II), Cd(II), Hg(II) and Bi(III) for the peptides show that Hg(II) has the highest affinity, owing to the initial formation of the extremely strong HgS₂ bond. Cd(II) and Pb(II) have comparable binding affinities at pH > 8, while Bi(III) displays the weakest affinity, following the model, M(II) + (TRI LXC)₃ ^3? → M(II)(TRI LXC)₃ ^?. While the relevant equilibria for Hg(II) binding to the TRI peptides corresponds to a strong first step forming Hg(TRI LXC)₂(HTRI LXC), followed by a single deprotonation to give Hg(TRI LXC)₃ ^?, the binding of Cd(II) and Pb(II) is consistent with initial formation of M(II)(TRI LXC)(HTRI LXC)₂ ⁺ at pH < 5 followed by a two-proton dissociation step (pK _a2) yielding M(II)(TRI LXC)₃ ^?. Pb(II)(TRI LXC)(HTRI LXC)₂ ⁺ converts to Pb(II)(TRI LXC)₃ ^? at slightly lower pH values than the corresponding Cd(II)–peptide complexes. In addition, Pb(II) displays a lower pK _a of binding to the “d”-substituted peptide, (TRI L12C, pK _a2 = 12.0) compared with the “a”-substituted peptide, (TRI L16C, pK _a2 = 12.6), the reverse of the order seen for Hg(II) and Cd(II). Pb(II) also showed a stronger binding affinity for TRI L12C (K _bind = 3.2 × 10⁷ M^?1) compared with that with TRI L16C (K _bind = 1.2 × 10⁷ M^?1) at pH > 8.     

Host-stage preference and functional response of aphid parasitoid Diaeretiella rapae (McIntosh) (Hym.: Braconidae) on greenbug,Schizaphis graminum (Rondani) (Hem: Aphididae)

Searching capacity is of great importance in the efficiency of natural enemies. In this research, the host-stage preference and functional response of Diaeretiella rapae (McIntosh) to different densities of greenbug, Schizaphis graminum (Rondani) were determined on Pishtaz wheat leaves at 25 ± 1°C, 60–70% relative humidity, and a photoperiod of 16:8 h (L:D). The host-stage preference was investigated by exposing 60 number of all nymphal instars and adults of greenbug to individual parasitoid females at eight replicates. Results indicated significant preference of D. rapae to second instar nymphs of S. graminum to perform functional response experiment. Densities of 2, 4, 8, 16, 32, 50, 64, 90 and 130 second instar nymphs of greenbugs were exposed to individual mated female wasps at 10 replicates for 24 h. Type of functional response was determined by logistic regression and its parameters, searching efficiency (a) and handling time (T _h), were estimated by non linear regression. The results indicated that the functional response of D. rapae was type III and values of searching efficiency, handling time and maximum rate of parasitism were calculated by two models as 0.1364 ± 0.084, 0.5761 ± 0.0179, 41.6594; 0.06489 ± 0.00652 (h^?1), 0.5628 ± 0.0173 (h) and 42.6439 (aphids/parasitoid/day), respectively.     

Investigation of the role of hydrophilic chain length in amphiphilic perfluoropolyether/poly(ethylene glycol) networks: towards high-performance antifouling coatings

The facile preparation of amphiphilic network coatings having a hydrophobic dimethacryloxy-functionalized perfluoropolyether (PFPE-DMA; M _w = 1500 g mol^?1) crosslinked with hydrophilic monomethacryloxy functionalized poly(ethylene glycol) macromonomers (PEG-MA; M _w = 300, 475, 1100 g mol^?1), intended as non-toxic high-performance marine coatings exhibiting antifouling characteristics is demonstrated. The PFPE-DMA was found to be miscible with the PEG-MA. Photo-cured blends of these materials containing 10 wt% of PEG-MA oligomers did not swell significantly in water. PFPE-DMA crosslinked with the highest molecular weight PEG oligomer (ie PEG1100) deterred settlement (attachment) of algal cells and cypris larvae of barnacles compared to a PFPE control coating. Dynamic mechanical analysis of these networks revealed a flexible material. Preferential segregation of the PEG segments at the polymer/air interface resulted in enhanced antifouling performance. The cured amphiphilic PFPE/PEG films showed decreased advancing and receding contact angles with increasing PEG chain length. In particular, the PFPE/PEG1100 network had a much lower advancing contact angle than static contact angle, suggesting that the PEG1100 segments diffuse to the polymer/water interface quickly. The preferential interfacial aggregation of the larger PEG segments enables the coating surface to have a substantially enhanced resistance to settlement of spores of the green seaweed Ulva, cells of the diatom Navicula and cypris larvae of the barnacle Balanus amphitrite as well as low adhesion of sporelings (young plants) of Ulva, adhesion being lower than to a polydimethyl elastomer, Silastic T2.     

M2X (M= Li,Na; X= F,Cl): the smallest superalkali clusters with significant NLO responses and electride characteristics

Hypervalent M₂X (M = Li, Na; X = F, Cl) clusters are prototype species possessing lower ionisation potentials than Li, therefore classified as superalkalis. This study reveals some interesting properties of these small clusters using ab initio MP2/aug-cc-pVTZ and QCISD/aug-cc-pVTZ methods. These clusters are shown as an ionic species, composed of positively charged cage of alkali metals (M₂⁺) and halogen anion (X^?). Therefore, the stability of M₂X is governed by both ionic and covalent interactions. We show that the excess valence electron of (M₂⁺) is pushed out by anionic X^?, which allows M₂X clusters to possess ‘electride’ characteristics. It is also due to this excess electron that M₂X clusters exhibit significant non-linear optical (NLO) properties. The dipole moment, mean polarisability and hyperpolarisability suggest their significant NLO responses, which are explained on the basis of electronic transitions in crucial excited states using TD-B3LYP/aug-cc-pVTZ method. The first static hyperpolarisabilities of Li₂F and Na₂F take the values of order of 10⁴ a.u. due to their lower transition energies. This study should provide new insights into the design of novel materials with significant NLO responses useful for electro-optical applications.     

The effects of (±)-, (+)-, and (−)-atenolol,sotalol, and amosulalol on the rat left atria and portal vein

The effects of (±)-, (+)-, and (?)-atenolol, sotalol, and amosulalol alone on the rat left atria and portal vein and on the respective β₁- and β₂-adrenoceptor-mediated responses to isoprenaline have been determined. (±)-Atenolol at 10^?6 M had no effect whereas high concentrations of (+)- and (?)-sotalol, 10^?5–10^?4 M, and (±)-, (+)-, and (?)-amosulalol depressed the response of the rat left atria to cardiac stimulation which indicates membrane stabilizing activity. None of the drugs tested had any effect alone on the rat portal vein. The order of potency as antagonists was (±)-amosulalol > (±)-atenolol > (±)-sotalol at β₁-adrenoceptors and (±)-amosulalol > (±)-sotalol > (±)-atenolol at β₂-adrenoceptors. (±)-Atenolol and (±)-amosulalol are β₁-selective whereas (±)-sotalol is β₂-selective. For each of the racemic β-blockers, the β₁- and β₂-adrenoceptor blocking activity was predominantly due to the (?)-enantiomer. © 1993 Wiley-Liss, Inc.     

Subcellular Localization and Kinetic Characterization of a Gill (Na+, K+)-ATPase from the Giant Freshwater Prawn Macrobrachium rosenbergii

The stimulation by Mg²⁺, Na⁺, K⁺, NH₄ ⁺, and ATP of (Na⁺, K⁺)-ATPase activity in a gill microsomal fraction from the freshwater prawn Macrobrachium rosenbergii was examined. Immunofluorescence labeling revealed that the (Na⁺, K⁺)-ATPase α-subunit is distributed predominantly within the intralamellar septum, while Western blotting revealed a single α-subunit isoform of about 108 kDa M _r. Under saturating Mg²⁺, Na⁺, and K⁺ concentrations, the enzyme hydrolyzed ATP, obeying cooperative kinetics with V _M = 115.0 ± 2.3 U mg^?1, K _0.5 = 0.10 ± 0.01 mmol L^?1. Stimulation by Na⁺ (V _M = 110.0 ± 3.3 U mg^?1, K _0.5 = 1.30 ± 0.03 mmol L^?1), Mg²⁺ (V _M = 115.0 ± 4.6 U mg^?1, K _0.5 = 0.96 ± 0.03 mmol L^?1), NH₄ ⁺ (V _M = 141.0 ± 5.6 U mg^?1, K _0.5 = 1.90 ± 0.04 mmol L^?1), and K⁺ (V _M = 120.0 ± 2.4 U mg^?1, K _M = 2.74 ± 0.08 mmol L^?1) followed single saturation curves and, except for K⁺, exhibited site–site interaction kinetics. Ouabain inhibited ATPase activity by around 73 % with K _I = 12.4 ± 1.3 mol L^?1. Complementary inhibition studies suggest the presence of F₀F₁–, Na⁺-, or K⁺-ATPases, but not V(H⁺)- or Ca²⁺-ATPases, in the gill microsomal preparation. K⁺ and NH₄ ⁺ synergistically stimulated enzyme activity (≈25 %), suggesting that these ions bind to different sites on the molecule. We propose a mechanism for the stimulation by both NH₄ ⁺, and K⁺ of the gill enzyme.     

Biological evaluation of a cytotoxic 2-substituted benzimidazole copper(II) complex: DNA damage,antiproliferation and apoptotic induction activity in human cervical cancer cells

Exploring novel chemotherapeutic agents is a great challenge in cancer medicine. To that end, 2-substituted benzimidazole copper(II) complex, [Cu(BMA)Cl₂]·(CH₃OH) (1) [BMA = N,N′-bis(benzimidazol-2-yl-methyl)amine], was synthesized and its cytotoxicity was characterized. The interaction between complex 1 and calf thymus DNA was detected by spectroscopy methods. The binding constant (K _b = 1.24 × 10⁴ M^?1) and the apparent binding constant (K _app = 6.67 × 10⁶ M^?1) of 1 indicated its moderate DNA affinity. Complex 1 induced single strand breaks of pUC19 plasmid DNA in the presence of H₂O₂ through an oxidative pathway. Cytotoxicity studies proved that complex 1 could inhibit the proliferation of human cervical carcinoma cell line HeLa in both time- and dose-dependent manners. The results of nuclei staining by Hoechst 33342 and alkaline single-cell gel electrophoresis proved that complex 1 caused cellular DNA damage in HeLa cells. Furthermore, treatment of HeLa cells with 1 resulted in S-phase arrest, loss of mitochondrial potential, and up-regulation of caspase-3 and -9 in HeLa cells, suggesting that complex 1 was capable of inducing apoptosis in cancer cells through the intrinsic mitochondrial pathway.     

Effective Cytochrome P450 (CYP) Inhibitors Isolated from Tarragon (Artemisia dracunculus)

Two effective cytochrome P450 (CYP) inhibitors were isolated from tarragon, Artemisia dracunculus. Their structures were spectroscopically identified as 2E,4E-undeca-2,4-diene-8,10-diynoic acid isobutylamide (1) and 2E,4E-undeca-2,4-diene-8,10-diynoic acid piperidide (2). Both compounds had dose-dependent inhibitory effects on CYP3A4 activity with IC₅₀ values of 10.0 ± 1.3 µM for compound 1 and 3.3 ± 0.2 µM for compound 2, and exhibited mechanism-based inhibition. This is the first reported isolation of effective CYP inhibitors from tarragon (Artemisia dracunculus) purchased from a Japanese market.     

A new sarasinoside congener,sarasinoside M2, from a marine sponge collected in the Solomon Islands

A new sarasinoside congener (sarasinoside M₂) and known sarasinoside B₁ were obtained from a marine sponge. Sarasinoside M₂ was suggested to have the same aglycon as sarasinoside M although the internal glucose in its sugar moiety is replaced by xylose. Sarasinosides B₁ and M₂ showed moderate cytotoxicity (approximate IC₅₀ 5–18 μM) toward Neuro-2a and HepG2 cell lines.     

A SUPEROXIDE DISMUTASE PURIFIED FROM THE ROOTS FROM Stemona tuberosa

Proteins from the fresh roots of Stemona tuberosa (Stemonaceae) were extracted into 20 mM phosphate buffer, pH 7.2/0.1 M NaCl, precipitated with 90% saturation ammonium sulfate, and enriched by diethylaminoethanol (DEAE) cellulose. The protein eluted as a single main peak from the unbound fractions (ST-1), and appeared as a single band with superoxide dismutase (SOD) activity after native polyacrylamide gel electrophoresis (PAGE) resolution and zymogram development. ST-1 was classified as SOD due to its strong inhibition by HCN and H₂O₂. The amino acid sequence of three tryptic peptides of ST-1 matched with the SOD isozymes from Ananas comosus and Solanum lycopersicum. The SOD consisted of at least two heterologous protein subunits with molecular mass of 17.6 and 31.5 kD, respectively, and had an optimal SOD activity at pH 5 and over a temperature range of 0–50°C. MgCl₂, MnCl₂, and HgCl₂ were strongly inhibitory at all concentrations tested. The SOD activity was completely negated in the presence of 0.5 mM SDS or 5 mM HgCl₂. The relationship between riboflavin and nitroblue tetrazolium (NBT) on SOD activity was linear, giving K _m and V _max values of the purified SOD of 62.414 ± 0.015 M and 101.010 ± 0.022 µmol/min/mg protein for NBT and 27.389 ± 0.032 M and 38.167 ± 0.021 µmol/min/mg protein for riboflavin, respectively.     

C-2 (E)-4-(Styryl)aniline substituted diphenylpyrimidine derivatives (Sty-DPPYs) as specific kinase inhibitors targeting clinical resistance related EGFRT790M mutant

With the aim to overcome the drug resistance induced by the EGFR T790M mutation (EGFR^T790M), herein, a family of diphenylpyrimidine derivatives (Sty-DPPYs) bearing a C-2 (E)-4-(styryl)aniline functionality were designed and synthesized as potential EGFR^T790M inhibitors. Among them, the compound 10e displayed strong potency against the EGFR^T790M enzyme, with the IC₅₀ of 11.0 nM. Compound 10e also showed a higher SI value (SI = 49.0) than rociletinib (SI = 21.4), indicating its less side effect. In addition, compound 10e could effectively inhibit the proliferation of H1975 cells harboring the EGFR^T790M mutation, within the concentration of 2.91 μM. Significantly, compound 10e has low toxicity against the normal HBE cell (IC₅₀ = 22.48 μM). This work provided new insights into the discovery of potent and selective inhibitor against EGFR^T790M over wild-type (EGFR^WT).