Dithiobis 2-nitrobenzoic Acid Research Articles

Isolation and functional characterization of two thioredoxin h isoforms from grape

Thioredoxins (Trxs) are small ubiquitous proteins that participate in dithiol-disulfide exchange reactions. In contrast to animals and prokaryotes, plants possess different types of Trxs that play a vital role in a number of different cellular processes. Two full-length cDNAs encoding different Trx h isoforms, designated VvTrx h2 and VvTrx h3, were isolated and cloned from grape (Vitis vinifera L. cv. Askari) berry tissue by rapid amplification of cDNA ends (RACE) method. VvTrx h2 and VvTrx h3 were heterologously expressed in Escherichia coli and their activities were compared using DTT-dependent insulin reduction and 5,5′-dithio-bis (2-nitrobenzoic acid) (DTNB) reduction activities. The NADPH-dependent DTNB reduction assay demonstrated that the both VvTrx h isoforms were reduced by NADPH-dependent thioredoxin reductase (NTR) from E. coli. Under heat shock treatment, the recombinant VvTrx h proteins formed the oligomeric structures at above 50 °C with a decrease in their disulfide reductase activities. The redox-dependent structural changes of VvTrx h2 and VvTrx h3 revealed that their oligomeric structures were changed into monomers and significantly increased their disulfide reductase activities. Furthermore, the both recombinant proteins were able to conserve a DTNB reduction activity even after 15 min heating at 99 °C.

Reporter Selection for Nanotags in Multiplexed Surface Enhanced Raman Spectroscopy Assays.

We report a quantitative evaluation of the choice of reporters for multiplexed surface-enhanced Raman spectroscopy (SERS). An initial library consisted of 15 reporter molecules that included commonly used Raman dyes, thiolated reporters, and other small molecules. We used a correlation matrix to downselect Raman reporters from the library to choose five candidates: 1,2-bis(4-pyridyl)ethylene, 4-mercaptobenzoic acid, 3,5-dichlorobenzenthiol, pentachlorothiophenol, and 5,5′-dithiobis(2-nitrobenzoic acid). We evaluated the ability to distinguish the five SERS reporters in a dipstick immunoassay for the biomarker human IgG. Raman nanotags, or gold nanostars conjugated to the five reporters and anti-human IgG polyclonal antibodies were constructed. A linear discriminant analysis approach was used to evaluate the separation of the nanotag spectra in mixtures of fixed ratios.

Glutathione Peroxidase Activity of Biological Samples Using A Novel Microplate-Based Method

Background: Glutathione peroxidase (GSH-Px) is one of the major enzyme in the antioxidative defense mechanism present in the cells. Methods: In the present study a novel and simple microplate-based method was developed, using the cupric neocuproine complex (Cu(Nc)2 2+) as a chromogenic oxidizing reagent, for the assessment of GSH-Px activity of biological samples for the first time. In this GSH-Px enzyme activity measurement, the GSH-Px catalyzed oxidation of Reduced Glutathione (GSH) gives rise to oxidized form of GSH (GSSG). The recommended method was based on the reduction of Cu(Nc)2 2+ to highly colored Cu(I)- neocuproine complex (Cu(Nc)2 +) by the unconsumed GSH, and measurement spectrophotometrically at 450 nm, the difference being correlated to GSH-Px activity of the analytes. Results: Under the optimum conditions, a linear calibration graph was obtained in the range of 18.5- 92.5 µM of GSH with Limit Of Detection (LOD) of 1.03 µM. Application of developed method to tissue homogenates (n = 9) provided GSH-Px activity values in agreement with those of the reference GSH-Px-5,5’-dithio-bis(2-nitrobenzoic acid) (DTNB) method. Conclusion: Cupric Ion Reducing Antioxidant Capacity (CUPRAC) antioxidant method was implemented in a microformat (96 well plates) and the reaction time of original CUPRAC method was significantly shortened from 30 to 4 min. Multi-sample can be simultaneously detected thanks to a novel microplate method. Keywords: GSH-Px activity, microplate based method, modified CUPRAC method, biological samples, neocuproine complex, reduced glutathione (GSH).

Cytosolic Diffusion and Peptide-Assisted Nuclear Shuttling of Peptide-Substituted Circa 102 Gold Atom Nanoclusters in Living Cells

For biological and medical applications, a definable nanomaterial and knowledge on its fate after administration are highly recommended if not mandatory. Here, we synthesized a water-soluble gold nanocluster by sodium borohydride reduction of chloroauric acid in the presence of 5,5′-dithio-bis(2-nitrobenzoic acid). The resulting gold nanocluster displayed the physical characteristics of a cluster containing an inner core of about 100 gold atoms that was surrounded by an organic monolayer made of about 30 thioaminobenzoic acids (TAB) and 14 anionic thionitrobenzoic acids (TNB). The mixed TAB-,TNB-protected gold nanocluster reacted well in water with thiolated peptides containing a nuclear localization signal (NLS) or a nuclear export signal (NES) mainly by exchange of the TNB, providing gold nanoclusters equipped with 8–9 intracellular active peptides and a remaining ligand coverage consisting mostly of the zwitterion TAB. The behavior of these peptide–gold nanoclusters inside the cytosol and nucleus of ce...

Selective Screening of Biological Thiols by Means of an Unreported Magenta Interaction and Evaluation Using Smartphones.

The determination of biological thiols is considered to be of great interest because of the participation of these molecules in physiological disorders as indicators of their existence and progress, such as Alzheimer’s disease, cancer, and cystic fibrosis, among others. Optical techniques for the quantification of these analytes are superior to other techniques in terms of selectivity, sensitivity, and cost. We describe here a simple protocol to determine biological thiols by means of a colorimetric analysis based on a magenta color interaction with 5,5′-dithiobis(2-nitrobenzoic acid). This methodology leverages the technology developed in smartphones, and it can be transferred to a simple analytical procedure through the use of absorbent strips and a cell phone. Excellent characteristics were observed, and this method opens up alternative avenues for the detection of biological thiols in a very simple and portable manner.

Determination of Blood Plasma Aminothiols Using Derivatization-enhanced Capillary Transient Isotachophoresis.

A sensitive capillary electrophoresis method was developed for the determination of aminothiol (cysteine, homocysteine, and glutathione) total levels in human blood plasma. Analytes were derivatized with Ellman's reagent (5,5'-dithiobis(2-nitrobenzoic acid)) after reduction with dithiothreitol. Liquid-liquid extraction was applied to purify the samples and concentrate the analytes. Total analysis time was 7.5 min using a silica capillary (50 μm i.d.; effective separation length 23.5 cm). Electrophoretic separation was performed using 50 mM citric acid with 20 mM triethanolamine (pH 3) containing 2% Ficoll 400. Detection limit was 0.8 μM for glutathione and 0.3 μM for both cysteine and homocysteine. Accuracy was 94 - 107%, repeatability and reproducibility were ca. 2.7 - 3.5 and 2.5 - 6.5%, respectively.

Stabilization of Butyrylcholinesterase by the Entrapment into the Natural Polymer-Based Gels.

A new method for obtaining stable butyrylcholinesterase (BuChE) samples based on the enzyme immobilization in starch and gelatin gels followed by drying is proposed. Coimmobilization of BuChE with the thiol group indicator 5,5'-dithiobis(2-nitrobenzoic) acid did not reduce the activity of BuChE, which allowed us to simplify the procedure and reduce the time of analysis of organophosphorus pesticides. The resulting immobilized samples retained activity for at least 300 days. BuChE samples based on the starch gel showed a greater sensitivity in the determination of pesticides as compared to the samples based on the gelatin gel.

In vitro anticholinesterase and neurotoxicity activities of Ocotea aciphylla fractions

Background: Ocotea species are known to produce secondary metabolites with a range of biological activities. This study aimed to evaluate the in vitro acetylcholinesterase (AChE) inhibition and neurotoxicity activities of the Ocotea aciphylla leaves. Materials and Methods: The in vitro anticholinesterase effect of crude extracts of O. aciphylla was investigated by means of spectrophotometric microplate assay. The most active extract, aqueous extract (AQE), was fractionated using column chromatography with silica gel as stationary phase to furnish several fractions that were also evaluated for the anticholinesterase effect. The neurotoxicity activity of AQE and active fraction (F9) was investigated in rat adrenal medulla pheochromocytoma strain cultures by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test. The chemical characterization of the most active fraction was performed through high-performance liquid chromatography coupled with mass spectrometry multistage (HPLC-MS/MS). Results: Ethanolic extract (EE) and AQE exhibited significant inhibitory effects of the activity of AChE, with inhibitory concentration (IC50) of 443.7 and 412.8 μg/mL, respectively. Among the fractions, F9 was more effective AChE inhibition with IC50of 286.2 μg/mL. In the neurocytotoxicity assays, only the F9, at the highest concentration (500 μg/mL), induced a significant reduction in cell viability. HPLC-MS/MS analysis of the active fraction enabled the characterization of the natural compounds, i.e., procyanidin B-type dimer, propelargonidin dimer, catechin, and methoxy-luteolin-deoxyhexose-hexose. Conclusion: The leaves of O. aciphylla showed in vitro anticholinesterase activity and low neurotoxicity, and these effects might be related to the presence of phenolic compounds. Abbreviation used: AChE: acetylcholinesterase; ACTI: Acetylthiocholine iodide; AD: Alzheimer's disease; AQE: Aqueous extract; BSA: Bovine serum albumin; DMSO: Dimethyl sulfoxide; DTNB: 5,5'-dithiobis (2-nitrobenzoic acid); EAE: Ethyl acetate extract; EE: Ethanolic extract; EtOAc: Ethyl acetate; HE: Hexane extract; Hex: Hexane; HPLC-MS/MS: High-performance liquid chromatography coupled with mass spectrometry multistage; MeOH: Methanol; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; PC-12: Rat adrenal medulla pheochromocytoma strain; RPMI: Roswell park memorial institute medium.

Assessment of Cysteine Reactivity of Human Hemoglobin at Its Residue Level: A Mass Spectrometry-Based Approach

In general, the reactivity of cysteine residues of proteins is measured by 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB) kinetics using spectrophotometry. Proteins with several cysteine residues may exhibit varying DTNB kinetics but residue level information can only be obtained with the prior knowledge of their three-dimensional structure. However, this method is limited in its application to the proteins containing chromophores having overlapping absorption profile with 2-nitro-5-thiobenzoic acid, such as hemoglobin (Hb). Additionally, this method is incapable of assigning cysteine reactivity at the residue levels of proteins with unknown crystal structures. However, a mass spectrometry (MS)-based platform might provide a solution to these problems. In the present study, alkylation kinetics of cysteine residues of adult human Hb (Hb A; α2β2) and sickle cell Hb (Hb S; HBB: c.20A>T) were investigated using matrix-assisted laser desorption/ionization (MALDI) MS. Differential site-specific reactivities of cysteine residues of Hb were investigated using alkylation kinetics with iodoacetamide (IAM). The observed reactivities corroborated well with the differential surface accessibilities of cysteine residues in the crystal structures of human Hb. The proposed method might be used to investigate cysteine reactivities of all the genetic and post-translational variants of Hb discovered to date. In addition, this method can be extended to explore cysteine reactivities of proteins, irrespective of the presence of chromophores and availability of crystal structures.

Toxicological profile and acetylcholinesterase inhibitory potential of Palicourea deflexa, a source of β-carboline alkaloids

Palicourea genus is chemically and taxonomically close to Psychotria genus, a well-known source of neuroactive alkaloids. It has been previously reported the pharmacological potential of these alkaloids in some targets related to the neurodegenerative process. In this context, this study was carried out in order to evaluate the toxic effects and acetylcholinesterase (AChE) inhibitory potential of Palicourea deflexa fraction of total alkaloids (FTA). P. deflexa FTA was analyzed by means of HPLC-DAD and HRMS-ESI. We performed toxicological screening through Fish Embryo Toxicity (FET) test using zebrafish embryo and abnormal developmental phenotypes were recorded daily. For AChE inhibition, zebrafish brains were used as enzymatic source and formation of thiolate dianion of 5,5′-Dithiobis(2-nitrobenzoic acid) (DTNB) was used to monitor acetylthiocholine hydrolysis. Lineaweaver–Burk double reciprocal plots were used to indicate mode of inhibition. Chemical analysis of the P. deflexa FTA allowed the identification of the main compound as harman-3-carboxylic acid. This fraction was evaluated in vivo for its toxicological effect. The zebrafish embryo test indicated that the FTA has a lethal concentration of 50% (LC50)=72.18μg/mL. Further, the FTA was evaluated for its AChE inhibitory profile, demonstrating an inhibitory concentration of 50% (IC50) of 50.65μg/mL. Lineaweaver–Burk double reciprocal plots indicated a mixed mode of inhibition. It is reported for the first time the toxicological and pharmacological profile of the alkaloid fraction of Palicourea deflexa in zebrafish models.

Physiopathology of multiple organ dysfunctions in severely monocrotophos-poisoned rabbits

ObjectiveAcute organophosphorus pesticides poisoning has a serious threat on people's health. This study aimed to investigate the pathogenesis and molecular mechanism of multiple organ dysfunction syndrome (MODS) in severely monocrotophos-poisoned rabbits. MethodsChinchilla rabbits were used to build the monocrotophos-poisoned animal model via subcutaneous abdominal injection. Acetylcholinesterase activity was determined using the dithiobisnitrobenzoic acid enzyme kinetics method, and the free organophosphorus (FOP) toxic substances content was analyzed using the enzyme inhibition method. The contents of tumor necrosis factor (TNF-α), interleukin 1-β (IL-β) and thromboxane B2 (TXB2) in the plasma and tissue homogenates were determined via radioimmunoassay. ResultsTwenty-four hours after exposure, in comparison to the plasma, blood cells and homogenates of various tissues, the bile had a significantly different FOP content (P < 0.05). In different phases, HE staining results confirmed that several degrees of pathological lesions (such as hemorrhage, edema, degeneration and necrosis) were detected in FOP poisoned rabbits. The TXB2 and TNF contents in plasma were significantly higher than those of the control (P < 0.05). Except for the intercostal muscle, all of the tissues had significantly higher TXB2 contents than the control. The TNF contents of the liver and lung and the IL-1β contents of the liver and kidney were significantly higher than those of the control (P < 0.05). ConclusionFOP stored in the gallbladder may play important role in enterohepatic circulation. In MODS rabbits, caused by OP poisoning, the TXB2 and TNF-α may play important role in inflammatory response and complement and coagulation systems respectively.

Advanced SERS Sensor Based on Capillarity-Assisted Preconcentration through Gold Nanoparticle-Decorated Porous Nanorods.

A preconcentrating surface-enhanced Raman scattering (SERS) sensor for the analysis of liquid-soaked tissue, tiny liquid droplets and thin liquid films without the necessity to collect the analyte is reported. The SERS sensor is based on a block-copolymer membrane containing a spongy-continuous pore system. The sensor's upper side is an array of porous nanorods having tips functionalized with Au nanoparticles. Capillarity in combination with directional evaporation drives the analyte solution in contact with the flat yet nanoporous underside of the SERS sensor through the continuous nanopore system toward the nanorod tips where non-volatile components of the analyte solution precipitate at the Au nanoparticles. The nanorod architecture increases the sensor surface in the detection volume and facilitates analyte preconcentration driven by directional solvent evaporation. The model analyte 5,5'-dithiobis(2-nitrobenzoic acid) can be detected in a 1 × 10-3 m solution ≈300 ms after the sensor is brought into contact with the solution. Moreover, a sensitivity of 0.1 ppm for the detection of the dissolved model analyte is achieved.

Exogenous hydrogen sulfide eliminates spatial memory retrieval impairment and hippocampal CA1 LTD enhancement caused by acute stress via promoting glutamate uptake

Acute stress impairs the hippocampus-dependent spatial memory retrieval, and its synaptic mechanisms are associated with hippocampal CA1 long-term depression (LTD) enhancement in the adult rats. Endogenous hydrogen sulfide (H2S) is recognized as a novel gasotransmitter and has the neural protective roles. However, very little attention has been paid to understanding the effects of H2S on spatial memory retrieval impairment. We observed the protective effects of NaHS (a donor of H2S) against spatial memory retrieval impairment caused by acute stress and its synaptic mechanisms. Our results showed that NaHS abolished spatial memory retrieval impairment and hippocampal CA1 LTD enhancement caused by acute stress, but not by glutamate transporter inhibitor l-trans-pyrrolidine-2,4-dicarboxylic (tPDC), indicating that the activation of glutamate transporters is necessary for exogenous H2S to exert its roles. Moreover, NaHS restored the decreased glutamate uptake in the hippocampal CA1 synaptosomal fraction caused by acute stress. Dithiothreitol (DTT, a disulfide reducing agent) abolished a decrease in the glutamate uptake caused by acute stress, and NaHS eradicated the decreased glutamate uptake caused by 5,5′-dithio-bis(2-nitrobenzoic)acid (DTNB, a thiol oxidizing agent), collectively, revealing that exogenous H2S increases glutamate uptake by reducing disulfide bonds of the glutamate transporters. Additionally, NaHS inhibited the increased expression level of phosphorylated c-Jun-N-terminal kinase (JNK) in the hippocampal CA1 region caused by acute stress. The JNK inhibitor SP600125 eliminated spatial memory retrieval impairment, hippocampal CA1 LTD enhancement and the decreased glutamate uptake caused by acute stress, indicating that exogenous H2S exerts these roles by inhibiting the activation of JNK signaling pathway.

Distigmine Bromide Produces Sustained Potentiation of Guinea-Pig Urinary Bladder Motility by Inhibiting Cholinesterase Activity.

Distigmine is a cholinesterase (ChE) inhibitor used for the treatment of detrusor underactivity in Japan. Distigmine's pharmacological effects are known to be long-lasting, but the duration of its effect on urinary bladder contractile function has not been fully elucidated. The present study aimed to determine these effects in relation to the plasma concentrations of distigmine and its inhibition of ChE activities in blood, plasma, and bladder tissue. Intravesical pressures were recorded in anesthetized guinea-pigs for 12 h after the intravenous administration of saline or distigmine (0.01-0.1 mg/kg). Plasma distigmine concentrations were measured by liquid chromatograph-tandem mass spectrometry (LC-MS/MS), while ChE activities were assayed using 5,5'-dithiobis(2-nitrobenzoic acid). Distigmine (0.1 mg/kg) significantly increased the maximum intravesical pressure at micturition reflex for 12 h post-administration. In contrast, plasma distigmine was only detectable for 6 h post-administration in these animals and a one-compartment model calculated an elimination half-life of 0.7 h. However, bladder and blood acetylcholinesterase activities were significantly inhibited for 12 h after distigmine administration, although plasma ChE activities were not affected. The pharmacodynamic effects of distigmine thus persisted after its elimination from the circulation, indicating that it may bind to bladder acetylcholinesterase, producing sustained enzyme inhibition and enhancement of bladder contractility.

In vivo anti-arthritic efficacy of Camellia sinensis (L.) in collagen induced arthritis model

Rheumatoid arthritis (RA), an autoimmune inflammatory disorder with synovial hyperplasia, destruction of cartilage, bone damage is often associated with risk of infections. Such risk could be attributed towards usage of immunosuppressive agents. Thus, the present study was undertaken to evaluate the anti-arthritic efficacy of aquo-alcoholic extract of Camellia sinensis (L.). Dried leaves of Camellia sinensis (L.) or Cs were filtered and extracted in 1:1 aqueous: ethanol by Soxhlet apparatus followed by lyophilization and spray drying to develop amorphous powder. Four different oral doses (50, 100, 200, 400mg/kg/body wt.) of aquo-alcoholic extract were evaluated for anti-edematogenic effect in collagen induced arthritis model. The selected anti-arthritic doses of Cs were evaluated for the oxidative stress markers like Glutathione [5-5'dithio-bis-2-nitrobenzoicacid (DTNB)], Superoxide dismutase [Epinephrine], Catalase [Hydrogen peroxide], Lipid peroxidation [Thiobarbituric acid reactive substance (TBARS)], Nitric oxide [Griess reagents:Nitrobluetetrazolium], Articular elastase [N-methoxysuccinyl-Ala-Ala-Pro- Val p-nitroanilide] in joints followed by haematological evaluation including RBC, WBC, Haemoglobin, platelets and haematocrit. To validate these biochemical changes, the radiological and histopathological (Haematoxylin & Eosin) evaluation was also conducted. The selected anti-arthritic dose of Cs i.e. 400mg/kg/body wt. (∼60% anti-arthritic efficacy on 35th day) could be attributed towards significant (p<0.05) increase in the levels of enzymatic (Superoxide dismutase and Catalase) and non-enzymatic (Glutathione) antioxidants by 34%, 59% and 50% respectively. Simultaneously, the significant (p<0.05) reduction of lipid peroxides, nitrite radical and elastase activity by 32%, 45% & 32% respectively as compare to control indicated overall decrease in oxidative stress. Haematological evaluation revealed restoration of RBC, WBC and platelets level in treatment group. The confirmatory analysis utilizing radiological and histological assessment showed alleviation of joint deformity, tissue swelling, pannus formation and neutrophils infiltration in treatment group as compared to collagen induced arthritis. The analysis showed that Cs can play an effective role in reduction of oxidative stress by modulating levels of antioxidants, reducing levels of free radicals while restoring normal haematopoietic cascade as observed in collagen induced arthritis model. Thus, the cumulative dose impact of 400mg/kg body wt., over a period of 14days also found extremely effective in terms of safeguarding their structural conformity against such auto-immune disorder.

The Effect of Liposomes on the Kinetics and Mechanism of the Photocatalytic Reduction of 5,5'-Dithiobis(2-Nitrobenzoic Acid) by Triethanolamine.

Recently, the addition of negatively charged liposomes was shown to increase the quantum yield of the photocatalytic reduction of 5,5'-dithio(2-nitrobenzoic acid) (H2DTNB) to 2-nitro-5-thiobenzoic acid (H2NTB) by triethanolamine using meso-tetra(4-(N-methylpyridinium)porphyrinato zinc chloride as photosensitizer. In this work, we investigate in detail the kinetics of this photocatalytic reaction both in homogeneous solution and at the surface of negatively charged liposomes, to unravel the effects of liposomes on the mechanism of the photoreaction. In homogeneous solution, the reaction is initiated by oxidative quenching. Both static (singlet) and dynamic (triplet) quenching of the photosensitizer contribute to the formation of the photoproduct. In these conditions, the reaction is limited by the low efficiency of reductive regeneration of the photosensitizer, compared to charge recombination. Upon adsorption of the positively charged photosensitizer to the negative surface of the liposomes, however, both static and dynamic oxidative quenching become ineffective due to electrostatic repulsion of the dianionic DTNB2- from the negatively charged membrane. In such conditions, photoreduction occurs via reductive quenching, showing that the addition of liposomes can truly modify the mechanism of photocatalyzed redox reactions.

5,5-Dithiobis(2-nitrobenzoic acid) pyrene derivative-carbon nanotube electrodes for NADH electrooxidation and oriented immobilization of multicopper oxidases for the development of glucose/O2 biofuel cells

We report the functionalization of multi-walled carbon nanotubes (MWCNTs) electrodes by a bifunctional nitroaromatic molecule accomplished via π-π interactions of a pyrene derivative. DTNB (5,5′-dithiobis(2-nitrobenzoic acid)) has the particularity to possess both electroactivable nitro groups and negatively charged carboxylic groups. The integration of the DTNB-modified MWCNTs was evaluated for different bioelectrocatalytic systems. The immobilized DTNB-based electrodes showed electrocatalytic activity toward the oxidation of the reduced form of nicotinamide adenine dinucleotide (NADH) with low overpotential of −0.09V vs Ag/AgCl at neutral pH. Glucose dehydrogenase was successfully immobilized at the surface of DTNB-based electrodes and, in the presence of NAD+, the resulting bioelectrode achieved efficient glucose oxidation with high current densities of 2.03mAcm−2. On the other hand, the aromatic structure and the negatively charged nature of the DTNB provoked orientation of both laccase and bilirubin oxidase onto the electrode, which enhanced their ability to undergo a direct electron transfer for oxygen reduction. Due to the proper orientation, low overpotentials were obtained (ca. 0.6V vs Ag/AgCl) and high electrocatalytic currents of about 3.5mAcm−2 were recorded at neutral pH in O2 saturated conditions for bilirubin oxidase electrodes. The combination of these bioanodes and bilirubin oxidase biocathodes provided glucose/O2 enzymatic biofuel cells (EBFC) exhibiting an open-circuit potential of 0.640V, with an associated maximum current density of 2.10mAcm−2. Moreover, the fuel cell delivered a maximum power density of 0.50mWcm−2 at 0.36 V.

A new Cd(II) coordination polymer constructed by a flexible carboxylate ligand

A new two-dimensional Cd(II) coordination polymer, [Cd(dtb)(CH3OH)] n (H 2 DTNB = 5,5′-dithiobis(2-nitrobenzoic acid), has been synthesized by solvothermal method, and characterized by elemental analysis, IR and single-crystal X-ray diffraction analysis. The complex crystallizes in the orthorhombic Pbca space group, with a = 15.1365(19) A, b = 7.6198(10) A, c = 31.761(4) A, β = 90°, V = 3663.2(8) A3, Z = 8, Mr = 538.77, D c = 1.954 g/cm3 and F(000) = 2128. The final R = 0.040 and wR = 0.0866 for 3603 observed reflections with I > 2σ(I). In the complex, the Cd(II) ion is coordinated with six oxygen atoms from four carboxylate groups of distinct DTNB ligands and one methanol molecule. The adjacent Cd centers are interconnected by the flexible carboxylate ligand to form a two-dimensional (2D) structure.

Tumoral gene silencing by receptor-targeted combinatorial siRNA polyplexes

Small interfering RNA (siRNA) promises high efficacy and excellent specificity to silence the target gene expression, which shows potential for cancer treatment. However, systemic delivery of siRNA with selectivity to the tumor site and into the cytosol of tumor cells remains a major limitation. To achieve this, we generated oligoaminoamide-based sequence-defined polycationic oligomers by solid-phase assisted synthesis, which can form polyplexes with anionic siRNA by electrostatic interaction to serve as siRNA carrier. Targeting for folate receptor (FR)-overexpressing tumors, we optimized the physicochemical properties of polyplexes by combinatorial optimization of PEGylated folate-conjugated oligomer (for FR targeting and shielding of surface charges) and 3-arm oligomer (for size modification and particle stability). For uni-directional fast coupling between the two groups of oligomers, we activated the cysteine thiol groups of one of the oligomers with 5,5′-dithio-bis(2-nitrobenzoic acid) to achieve a fast chemical linkage through disulfide formation with the free thiol groups of the other oligomer. These targeted combinatorial polyplexes (TCPs) are homogeneous spherical particles with favorable size and surface charge, which showed strong siRNA binding activity. TCPs were internalized into cells by FR-mediated endocytosis, triggered significant eGFP-luciferase marker gene silencing, and transfection with antitumoral EG5 siRNA suppressed cell proliferation in FR-expressing tumor cells. Moreover, the most promising formulation TCP1 after i.v. administration in tumor-bearing mice exhibited siRNA delivery into the tumor, resulting in EG5 gene silencing at mRNA level. Therefore, by covalent combination of two sequence-defined functional oligomers, we developed a siRNA carrier system with optimized size and surface charge for efficient tumor cell-directed gene silencing and cytotoxicity in vitro and in vivo.

Surface Enhanced Raman Spectroscopy Sensor Based on Magnetic Beads-induced Nanoparticles Aggregation for Detection of Bacterial Deoxyribonucleic Acid

Abstract A novel method was developed for the highly sensitive detection of bacterial DNA by surface-enhanced Raman spectroscopy (SERS) with 5,5′-dithio-bis(2-nitrobenzoic acid) (DTNB)-modified gold nanoparticles as reporter probes and on the basis of the separation and enrichment effect of magnetic beads and the high affinity of fully complementary hybridization of DNA strands. Capture probe was immobilized onto the surface of Streptavidin-enwrapped magnetic beads (SA-MB) through the force between biotin and avidin, which hybridized with the target bacterial DNA sequence partly, and the remaining connected with the report probe decorated AuNPs with DTNB and SH-DNA (AuNPs@DTNB@DNA). Compared with previous methods, this design shortened the distance between particles by the aggregation effect of the magnetic beads to nanoparticles, and produced the plasma resonance coupling effect, which increased the SERS signal significantly. The results showed that, under the optimized conditions, the concentration of DNA in the range from 5 pM to 5 nM performed a good linear relationship with Raman intensity. The limit of detection (LOD) for bacterial DNA was estimated to be about 5 pM, showing a certain selectivity. The method was simple in design with low cost, also it was used in the sensitive and selective detection of bacterial DNA.