Optimization of wavefront reconstruction accuracy for conjugate shift differential absolute testing.

The conjugate shift differential method, based on Fourier transforms, is critical for surface error testing of high-precision optical elements. However, this common approach is also prone to periodic spectrum loss. As such, this paper proposes conjugate double shift differential (CDSD) absolute testing, which can effectively compensate for spectrum loss and achieve accurate wavefront reconstructions. Spectrum loss in the single shift differential method is analyzed through a study of the Fourier reconstruction process. A calculation model for the proposed CDSD method is then established and constraint conditions for shift quantities are provided by analyzing double shear effects observed in transverse shear interference. Finally, the reconstruction accuracies of various spectrum compensation methods are compared. Results showed that spectrum loss became more evident with increasing shift amounts. However, the CDSD method produced the smallest measurement error compared with conventional direct zero filling and adjacent point averaging, suggesting our approach could effectively improve absolute shape measurement accuracy for planar optical elements.

Gold nanoparticle-based colorimetric aptasensor for rapid detection of multiple mycotoxins in rice.

A novel, simple and rapid colorimetric aptasensor for multiple mycotoxins (ochratoxin A (OTA) and aflatoxin B1 (AFB1)) detection was developed using unmodified gold nanoparticles (AuNPs). In the work, the high affinities of OTA and AFB1 aptamers were employed as the recognition elements for the colorimetric determination of OTA and AFB1. In the absence of mycotoxins, the sulfhydryl-modified aptamers were directly adsorbed to the AuNP surface through Au-S bonds, further prohibiting the aggregation induced by a high concentration of salt, and the solutions remain red. In the presence of mycotoxins, the corresponding aptamer-target complexes were formed and the corresponding aptamers were detached from the surface of AuNPs, leading to the aggregation of AuNPs under the optimal salt solution and a color change. By spectroscopic quantitative analysis and visual analysis, the LODs of OTA and AFB1 were down to 0.005 ng mL-1 and 0.07 ng mL-1, respectively. Furthermore, the colorimetric aptasensor showed a high specificity in the presence of other interfering mycotoxins and metal ions. Finally, the developed aptasensor was applicable to detect OTA and AFB1 in rice samples with satisfactory performance. Our strategy has great potential for the rapid and sensitive detection of OTA and AFB1 for on-site analysis.

SMS regulates the expression and function of P-gp and MRP2 in Caco-2 cells.

Sphingomyelin synthase (SMS) has two isoforms of SMS1 and SMS2, the last enzyme involved in the biosynthesis of sphingomyelin (SM), and has impact on the expression of membrane proteins. In the present study, we explored the potential effects of SMS on drug transporters, a special family of membrane proteins in human intestinal epithelial Caco-2 cells. The specific knockdown of SMS1 or SMS2 with siRNA in Caco-2 cells substantially decreased the expression and function of P-glycoprotein (P-gp) and multidrug resistance protein 2 (MRP2) rather than other drug transporters MRP1, MRP3, PEPT1, OATP2B1, and BCRP. In the SMS1 stable overexpressed Caco-2 cell line, the expression levels of P-gp and MRP2 and transcription factor pregnane X receptor (PXR) were upregulated and the phosphorylation levels of signaling pathways janus protein tyrosine kinase 2 (JAK-2) and extracellular signal-regulated kinases (ERK) were also evidently increased; however, the upregulated mRNA expression levels of PXR, P-gp, and MRP2 were diminished by inhibiting the phosphorylation of ERK and JAK-2. Furthermore, the SMS1 overexpression in Caco-2 cells altered the expression levels of ERM proteins ezrin and moesin, which are closely connected to the function of drug transporters. In conclusion, we herein demonstrate for the first time that in Caco-2 cells SMS regulates the expression and function of drug transporters P-gp and MRP2, and their regulator PXR is mediated by phosphorylated ERK and JAK-2 signaling pathways.

Oral delivery of Angiotensin-converting enzyme 2 and Angiotensin-(1-7) bioencapsulated in plant cells attenuates pulmonary hypertension.

Emerging evidences indicate that diminished activity of the vasoprotective axis of the renin-angiotensin system, constituting angiotensin-converting enzyme 2 (ACE2) and its enzymatic product, angiotensin-(1-7) [Ang-(1-7)] contribute to the pathogenesis of pulmonary hypertension (PH). However, long-term repetitive delivery of ACE2 or Ang-(1-7) would require enhanced protein stability and ease of administration to improve patient compliance. Chloroplast expression of therapeutic proteins enables their bioencapsulation within plant cells to protect against gastric enzymatic degradation and facilitates long-term storage at room temperature. Besides, fusion to a transmucosal carrier helps effective systemic absorption from the intestine on oral delivery. We hypothesized that bioencapsulating ACE2 or Ang-(1-7) fused to the cholera nontoxin B subunit would enable development of an oral delivery system that is effective in treating PH. PH was induced in male Sprague Dawley rats by monocrotaline administration. Subset of animals was simultaneously treated with bioencapsulaed ACE2 or Ang-(1-7) (prevention protocol). In a separate set of experiments, drug treatment was initiated after 2 weeks of PH induction (reversal protocol). Oral feeding of rats with bioencapsulated ACE2 or Ang-(1-7) prevented the development of monocrotaline-induced PH and improved associated cardiopulmonary pathophysiology. Furthermore, in the reversal protocol, oral ACE2 or Ang-(1-7) treatment significantly arrested disease progression, along with improvement in right heart function, and decrease in pulmonary vessel wall thickness. In addition, a combination therapy with ACE2 and Ang-(1-7) augmented the beneficial effects against monocrotaline-induced lung injury. Our study provides proof-of-concept for a novel low-cost oral ACE2 or Ang-(1-7) delivery system using transplastomic technology for pulmonary disease therapeutics.

Modulation of intestinal folate absorption by erythropoietin in vitro.

Besides the direct stimulation of erythropoiesis, erythropoietin (EPO) therapy in renal anemia may also play a regulatory role in maintaining the homeostasis of hematopoietic nutrients. It has been reported that EPO can stimulate intestinal iron absorption. However, the involvement of EPO in intestinal folate absorption remains elusive. The objective of this study was to investigate the effect of EPO on intestinal transport of folate in vitro and to elucidate the possible mechanism(s) involved in this regulation. Transport assays of folic acid were performed in Caco-2 monolayers treated with EPO. The effect of EPO on the expression of transporters involved in the folate absorption was investigated. The possible involvement of three main EPO signaling pathways, the janus protein tyrosine kinase 2 (JAK-2) pathway, extracellular signal regulated kinases (ERK) pathway, and phosphatidylinositol 3 kinase/Akt (PI3K/Akt) pathway, in the transporter regulation was explored. The absorptive flux (apical to basolateral) of folic acid was enhanced by EPO treatment in a dose-dependent manner, which was companied with the significant up-regulation of reduced folate carrier (RFC) and apical proton coupled folate transporter (PCFT). The efflux (basolaterial to apical) of folic acid was enhanced only by the high dose of EPO treatment, which was associated with the significant up-regulation of apical multidrug resistance-associated protein 2 (MRP2). The expression levels of all of these transporters were up-regulated by EPO treatment in a dose- and time-dependent manner. Transporter expression in response to blocking EPO induced activation of JAK-2, ERK, and PI3K/Akt was changed to a different extent. As a conclusion, intestinal folate absorption was enhanced by EPO treatment in vitro. Our findings provided direct evidence to establish the correlation between EPO and folate homeostasis.

Sensitive determination of buformin using poly-aminobenzoic acid modified glassy carbon electrode.

Glassy carbon electrode, which is used to electrochemically determine the content of buformin, is modified with an electropolymerized film of p-aminobenzoic acid in pH 7.0 acetate buffer solution (ABS). The polymer showed an excellent electrocatalytic activity for the reduction of buformin. In pH 7.0 ABS, the cathodic peak current increased linearly over three concentration intervals of buformin, and the detection limit (S/N=3) was 2.0×10-9 g/mL. The method was successfully applied to directly determine buformin in tablets with standard addition recoveries of 95.8-102.5%. The proposed method is simple, cheap and highly efficient.

Highly sensitive and reproducible cyclodextrin-modified gold electrodes for probing trace lead in blood.

A highly sensitive and reproducible lead sensor based on a cyclodextrin-modified gold electrode was created. A self-assembled monolayer (SAM) of thiolated beta-cyclodextrin (6-(2-mercapto-ethylamino)-6-deoxy-beta-cyclodextrin (MEA-beta-CD)) was prepared and modified on a gold electrode (MCGE) for specific Pb(2+)-sensing. Thus the mercury-free sensors for Pb(2+) assay based on MCGE were established. A linear calibration response for Pb(2+) was found in the range of 1.7 x 10(-8)M to 9.3 x 10(-7)M. The detection limit was 7.1 x 10(-9)M (with S/N>3), which was 10 times lower than other reported methods of detection Pb(2+) with CD. The measurement results via this method for real blood samples were well agree with those obtained by ICP-AES, and thus presented a novel strategy in design of specific lead sensors with high sensitivity and stability for analysis of trace Pb(2+) in real blood samples.

Novel superparamagnetic core-shell molecular imprinting microspheres towards high selective sensing.

Novel superparamagnetic core-shell imprinting microspheres (MCSIMs) were synthesized using magnetite microspheres with 350 nm diameter and 70 nm thickness silica gel to form core-shell Fe(3)O(4)/SiO(2) composite for template phenylephrine (Phen) recognition and high efficiency separation. Compared to the previous imprinting recognition, the main advantage of this strategy lies in two aspects: one is the high stability and monodispersity of the MCSIMs structure, the other is the use of superparamagnetic Fe(3)O(4)/SiO(2) microspheres as an immobilization matrix and separation tool, thus greatly simplifying time-consuming washing steps. The affinity and selectivity of the MCSIMs were monitored by QCM and electrochemistry measurements. Imprinting microspheres have a remarkable affinity to Phen over that of structurally related molecules, including DA, EP, Phe and Tyr. The relative binding selectivity for different analytes estimated from amperometric signals was Phen : DA : EP = 40 : 5 : 1. The MCSIMs sensor showed a high sensitivity (400 microA mM(-1)), short response time (reaching 98% within 10 s), and broad linear response range from 1 microM to 0.1 mM and low detection limit (0.1 microM). Additionally, the results of control experiments showed that only negligible signal was obtained for non-imprinting microspheres. This could be reasonably attributed to the unique surface pores, charges and especially the nature of the functional groups inside MCSIMs cavities.

Novel renewable immunosensors based on temperature-sensitive PNIPAAm bioconjugates.

A novel renewable immunosensor was created comprising a temperature-controlled surface composed of poly(n-isopropylacrylamide) (PNIPAAm)-antibody conjugates that could reversibly bind the antigen. Bovine serum albumin (BSA) and the corresponding antibody (anti-BSA) were chosen as a model antibody-antigen system to demonstrate the concept. The thermally responsive PNIPAAm conjugated to anti-BSA displayed a controllable conformation change between an expanded and a collapsed form, below and above its characteristic phase transition temperature, i.e. low critical solution temperature (LCST). This showed a remarkable change in the bioaffinity of the conjugate for BSA. Thus, a renewable anti-BSA surface was generated for re-binding of the target antigen at the thermally controllable PNIPAAm-anti-BSA conjugated surface. The temperature-controlling strategy resulted in the regeneration of immunosensors on which immobilized anti-BSA antibodies retained their activity and specificity for more than 30 reproducible assays. The level of dissociation reached 89%, which is comparable with established recovery methods, while offering easer handing. The controlled binding and dissociation were monitored by quartz crystal microbalance (QCM), confocal fluorescence, native electrophoresis, laser-induced fluorescence, and electrochemical impedance methods.

Sensitive detection of trifluoperazine using a poly-ABSA/SWNTs film-modified glassy carbon electrode.

A poly-ABSA/SWNTs composite-modified electrode was fabricated by electropolymerizing aminobenzene sulphonic acid (ABSA) on the surface of glassy carbon electrode (GCE) modified with single-wall carbon nanotubes (SWNTs). SWNTs provide a 3D porous and conductive network for the polymer immobilization. The nanocomposite film was characterized by scanning electron microscope (SEM) and electrochemical impedance spectroscopy (EIS). The results indicated that this composite-modified electrode had strong electrocatalytic activity toward the oxidation of trifluoperazine (TFP). TFP could effectively accumulate on the modified electrode and generate a sensitive anodic peak at 0.72V (versus SCE) in pH 6.1 phosphate buffer solution. Under the selected conditions, the anodic peak current of TFP was linear with its concentration within the range from 1.0x10(-7) to 1.0x10(-5)molL(-1) and 1.0x10(-5) to 1.0x10(-4)molL(-1), and the detection limit was 1.0x10(-9)molL(-1) (S/N=3). This method was successfully applied to the detection of trifluoperazine in drug samples and the recovery was satisfactory. In comparison with the SWNTs/GCE or poly-ABSA/GCE prepared in the similar way, this composite-modified electrode exhibited better catalytic activity.

Sensitive determination of phenylephrine and chlorprothixene at poly(4-aminobenzene sulfonic acid) modified glassy carbon electrode.

Phenylephrine (i.e. PHE) and chlorprothixene (i.e. CPT), two effective and important antipsychotic drugs with low redox activity, were found generating an irreversible anodic peak at about +0.89 V (vs. SCE) and +1.04 V in 0.05 M HAc-NaAc (pH 5.0) or NH(2)CH(2)COOH-HCl (pH 2.4) buffer solution at poly(4-aminobenzene sulfonic acid) modified glassy carbon electrode (i.e. poly(4-ABSA)/GC), respectively. Sensitive and quantitative measurement for them based on the anodic peaks was established under the optimum conditions. The anodic peak current was linear to PHE and CPT concentrations from 1 x 10(-7) to 1.5 x 10(-5)M and 2 x 10(-6) to 4.5 x 10(-5)M, the detection limits obtained were 1 x 10(-8) and 1 x 10(-7)M, separately. The modified electrode exhibited some excellent characteristics including easy regeneration, high stability, good reproducibility and selectivity. The method proposed was successfully applied to the determination of PHE and CPT in drug injections or tablets and proved to be reliable compared with ultraviolet spectrophotometry. The modified electrode was characterized by electrochemical methods.

Sensitive Detection of Haloperidol and Hydroxyzine at Multi-Walled Carbon Nanotubes-Modified Glassy Carbon Electrodes.

Haloperidol (i.e. HPD) and hydroxyzine (i.e. HXY), two effective and important tranquilizers with low redox activity, were found to generate an irreversible anodic peak at about +0.86 V (vs. SCE) or two anodic peaks at about +0.83 and +0.91 V in 0.05 M NaH2PO4-Na2HPO4 (pH=7.0) buffer solution with a multi-walled carbon nanotubes-modified glassy carbon electrode (i.e. MWNTs/GC), respectively. Their sensitive and quantitative measurement based on the first two anodic peaks was established under the optimum conditions. The anodic peak current was linear to HPD and HXY concentration from 1×10-7 to 2.5 ×10-5 M and 5×10-8 to 2.5 ×10⁻5 M, the detection limits obtained were 8×10-9 and 5×10-9 M, separately. The modified electrode exhibited some excellent characteristics including easy regeneration, high stability, good reproducibility and selectivity. The method proposed was successfully applied to the detection of HPD and HXY in drug tablets and proved to be reliable compared with ultraviolet spectrophotometry. The modified electrode was characterized by electrochemical methods.

Poly (O-aminobenzoic acid) modified glassy carbon electrode for electrochemical detection of dopamine in the presence of ascorbic acid.

O-aminobenzoic acid (o-ABA) film is deposited on glassy carbon electrode (GCE) by electropolymerization in pH 7.0 phosphate buffer solution (PBS). The polymeric film shows an excellent electrocatalytical activity on the oxidation of dopamine (DA). Difference pulse voltammetry (DPV) was performed to determine DA in an excess of ascorbic acid (AA). The oxidation peak potentials of DA and AA recorded are 144 mV and -52 mV, respectively. In pH 7.0 PBS, the anodic peak current of DA increases linearly over two concentration intervals, viz., 1.0x10(-7)-1.0x10(-5) mol L(-1) and 1.0x10(-5) - 2.0x10(-4) mol L(-1), with correlation coefficient, 0.9966 and 0.9960, respectively. The relative standard deviation of 10 successive scans is 2.8 % for 1.0x10(-6) mol L(-1) DA and the recovery is 96 % - 101 %. The interference of AA and DOPAC with the determination of DA could be eliminated because of the very distinct attracting interaction between DA cations and the negatively poly (o-ABA) film in pH 7.0 PBS. The proposed method exhibits good recovery and reproducibility.