Fiber Optic Localized Surface Plasmon Resonance Sensor Based on Carboxymethylated Dextran Modified Gold Nanoparticles Surface for High Mobility Group Box 1 (HMGB1) Analysis.

Rapid, sensitive, and reliable detection of high mobility group box 1 (HMGB1) is essential for medical and diagnostic applications due to its important role as a biomarker of chronic inflammation. Here, we report a facile method for the detection of HMGB1 using carboxymethyl dextran (CM-dextran) as a bridge molecule modified on the surface of gold nanoparticles combined with a fiber optic localized surface plasmon resonance (FOLSPR) biosensor. Under optimal conditions, the results showed that the FOLSPR sensor detected HMGB1 with a wide linear range (10-10 to 10-6 g/mL), fast response (less than 10 min), and a low detection limit of 43.4 pg/mL (1.7 pM) and high correlation coefficient values (>0.9928). Furthermore, the accurate quantification and reliable validation of kinetic binding events measured by the currently working biosensors are comparable to surface plasmon resonance sensing systems, providing new insights into direct biomarker detection for clinical applications.

Determination of the Highly Sensitive Carboxyl-Graphene Oxide-Based Planar Optical Waveguide Localized Surface Plasmon Resonance Biosensor.

This study develops a highly sensitive and low-cost carboxyl-graphene-oxide-based planar optical waveguide localized surface plasmon resonance biosensor (GO-OW LSPR biosensor), a system based on measuring light intensity changes. The structure of the sensing chip comprises an optical waveguide (OW)-slide glass and microfluidic-poly (methyl methacrylate) (PMMA) substrate, and the OW-slide glass surface-modified gold nanoparticle (AuNP) combined with graphene oxide (GO). As the GO has an abundant carboxyl group (-COOH), the number of capture molecules can be increased. The refractive index sensing system uses silver-coated reflective film to compare the refractive index sensitivity of the GO-OW LSPR biosensor to increase the refractive index sensitivity. The result shows that the signal variation of the system with the silver-coated reflective film is 1.57 times that of the system without the silver-coated reflective film. The refractive index sensitivity is 5.48 RIU-1 and the sensor resolution is 2.52 ± 0.23 × 10-6 RIU. The biochemical sensing experiment performs immunoglobulin G (IgG) and streptavidin detection. The limits of detection of the sensor for IgG and streptavidin are calculated to be 23.41 ± 1.54 pg/mL and 5.18 ± 0.50 pg/mL, respectively. The coefficient of variation (CV) of the repeatability experiment (sample numbers = 3) is smaller than 10.6%. In addition, the affinity constants of the sensor for anti-IgG/IgG and biotin/streptavidin are estimated to be 1.06 × 107 M-1 and 7.30 × 109 M-1, respectively. The result shows that the GO-OW LSPR biosensor has good repeatability and very low detection sensitivity. It can be used for detecting low concentrations or small biomolecules in the future.

Integrated Graphene Oxide with Noble Metal Nanoparticles to Develop High-Sensitivity Fiber Optic Particle Plasmon Resonance (FOPPR) Biosensor for Biomolecules Determination.

In this research, a direct, simple and ultrasensitive fiber optic particle plasmon resonance (FOPPR) biosensing platform for immunoglobulin G (IgG) detection was developed using a gold nanoparticle/graphene oxide (AuNP/GO) composite as signal amplification element. To obtain the best analytical performance of the sensor, experimental parameters including the surface concentration of GO on the AuNPs, formation time of the GO, the concentration of the anti-IgG and incubation time of anti-IgG were optimized. The calibration plots displayed a good linear relationship between the sensor response (ΔI/I0) and the logarithm of the analyte concentrations over a linear range from 1.0 × 10-10 to 1.0 × 10-6 g/mL of IgG under the optimum conditions. A limit of detection (LOD) of 0.038 ng/mL for IgG was calculated from the standard calibration curve. The plot has a linear relationship (correlation coefficient, R = 0.9990). The analytical performance of present work's biosensor was better than that of our previously reported mixed self-assembled monolayer of 11-mercaptoundecanoic acid/6-mercapto-1-hexanol (MUA/MCH = 1:4) method by about three orders of magnitude. The achieved good sensitivity may be attributed to the synergistic effect between GO and AuNPs in this study. In addition, GO could immobilize more antibodies due to the abundant carboxylic groups on its surface. Furthermore, we also demonstrated that the results from this sensor have good reproducibility, with coefficients of variation (CVs) < 8% for IgG. Therefore, the present strategy provides a novel and convenient method for chemical and biochemical quantification and determination.

Fiber Optic Particle Plasmon Resonance-Based Immunoassay Using a Novel Multi-Microchannel Biochip.

A novel multi-microchannel biochip fiber-optic particle plasmon resonance (FOPPR) sensor system for the simultaneous detection of multiple samples. The system integrates a novel photoelectric system, a lock-in module, and an all-in-one platform incorporating optical design and mechanical design together to improve system stability and the sensitivity of the FOPPR sensor. The multi-microchannel FOPPR biochip has been developed by constructing a multi-microchannel flow-cell composed of plastic material to monitor and analyze five samples simultaneously. The sensor system requires only 30 µL of sample for detection in each microchannel. Moreover, the total size of the multi-microchannel FOPPR sensor chip is merely 40 mm × 30 mm × 4 mm; thus, it is very compact and cost-effective. The analysis was based on calibration curves obtained from real-time sensor response data after injection of sucrose solution, streptavidin and anti-dinitrophenyl (anti-DNP) antibody of known concentrations over the chips. The results show that the multi-microchannel FOPPR sensor system not only has good reproducibility (coefficient of variation (CV) < 10%), but also excellent refractive index resolution (6.23 ± 0.10 × 10-6 refractive index unit (RIU)). The detection limits are 2.92 ± 0.28 × 10-8 g/mL (0.53 ± 0.01 nM) and 7.48 ± 0.40 × 10-8 g/mL (0.34 ± 0.002 nM) for streptavidin and anti-DNP antibody, respectively.

Effect of Surface Coverage of Gold Nanoparticles on the Refractive Index Sensitivity in Fiber-Optic Nanoplasmonic Sensing.

A simple theoretical model was developed to analyze the extinction spectrum of gold nanoparticles (AuNPs) on the fiber core and glass surfaces in order to aid the determination of the surface coverage and surface distribution of the AuNPs on the fiber core surface for sensitivity optimization of the fiber optic particle plasmon resonance (FOPPR) sensor. The extinction spectrum of AuNPs comprises of the interband absorption of AuNPs, non-interacting plasmon resonance (PR) band due to isolated AuNPs, and coupled PR band of interacting AuNPs. When the surface coverage is smaller than 12.2%, the plasmon coupling effect can almost be ignored. This method is also applied to understand the refractive index sensitivity of the FOPPR sensor with respect to the non-interacting PR band and the coupled PR band. In terms of wavelength sensitivity at a surface coverage of 18.6%, the refractive index sensitivity of the coupled PR band (205.5 nm/RIU) is greater than that of the non-interacting PR band (349.1 nm/RIU). In terms of extinction sensitivity, refractive index sensitivity of the coupled PR band (-3.86/RIU) is similar to that of the non-interacting PR band (-3.93/RIU). Both maximum wavelength and extinction sensitivities were found at a surface coverage of 15.2%.

Self-referencing fiber optic particle plasmon resonance sensing system for real-time biological monitoring.

We present the design and experimental verification of a self-referencing dual-channel fiber optic particle plasmon resonance (FOPPR) sensing system for compensation of thermal and bulk-composition effects as well as nonspecific adsorption in real-time biosensing of complex samples. A theoretical model is first proposed and then a systematic experimental approach is used to verify the model. The sensing system comprises an analysis fiber sensor and a reference fiber sensor in a single microfluidic chip, where the analysis fiber is functionalized with a recognition molecule. The compensation still works even if the surface coverages of gold nanoparticles on the reference and analysis fibers are not exactly the same. The potential of this approach is illustrated by a model biosensing experiment in which the detection of anti-biotin is compensated for bulk refractive index change, nonspecific adsorption and/or color interference, in various sample media. The percent recovery is 103.2% under both the effects of bulk refractive index change and nonspecific adsorption and is 93.9% under both the effects of color interference and nonspecific adsorption, suggesting that the compensation is effective.

Fabrication quality analysis of a fiber optic refractive index sensor created by CO2 laser machining.

This study investigates the CO2 laser-stripped partial cladding of silica-based optic fibers with a core diameter of 400 µm, which enables them to sense the refractive index of the surrounding environment. However, inappropriate treatments during the machining process can generate a number of defects in the optic fiber sensors. Therefore, the quality of optic fiber sensors fabricated using CO2 laser machining must be analyzed. The results show that analysis of the fiber core size after machining can provide preliminary defect detection, and qualitative analysis of the optical transmission defects can be used to identify imperfections that are difficult to observe through size analysis. To more precisely and quantitatively detect fabrication defects, we included a tensile test and numerical aperture measurements in this study. After a series of quality inspections, we proposed improvements to the existing CO2 laser machining parameters, namely, a vertical scanning pathway, 4 W of power, and a feed rate of 9.45 cm/s. Using these improved parameters, we created optical fiber sensors with a core diameter of approximately 400 µm, no obvious optical transmission defects, a numerical aperture of 0.52 ± 0.019, a 0.886 Weibull modulus, and a 1.186 Weibull-shaped parameter. Finally, we used the optical fiber sensor fabricated using the improved parameters to measure the refractive indices of various solutions. The results show that a refractive-index resolution of 1.8 × 10(-4) RIU (linear fitting R2 = 0.954) was achieved for sucrose solutions with refractive indices ranging between 1.333 and 1.383. We also adopted the particle plasmon resonance sensing scheme using the fabricated optical fibers. The results provided additional information, specifically, a superior sensor resolution of 5.73 × 10(-5) RIU, and greater linearity at R2 = 0.999.

Formation of amyloid fibrils from ß-amylase.

Fibril formation has been considered a significant feature of amyloid proteins. However, it has been proposed that fibril formation is a common property of many proteins under appropriate conditions. We studied the fibril formation of ß-amylase, a non-amyloid protein rich in α-helical structure, because the secondary structure of ß-amylase is similar to that of prions. With the conditions for the fibril formation of prions, ß-amylase proteins were converted into amyloid fibrils. The features of ß-amylase proteins and fibrils are compared to prion proteins and fibrils. Furthermore, the cause of neurotoxicity in amyloid diseases is discussed.

Schisandrin enhances dendrite outgrowth and synaptogenesis in primary cultured hippocampal neurons.

BACKGROUND: Schisandra chinensis, commonly used in Asia for tea material and traditional Chinese medicine, is presumed to enhance mental and intellectual functions. In this study, the effects and signalling mechanisms of a purified compound schisandrin, one of the lignan of Schisandra chinensis, on primary cultured hippocampal neurons were investigated. RESULTS: Schisandrin treatment enhanced total dendritic length and branching complexity, both of which were significantly suppressed in the presence of specific blockers for calmodulin-dependent kinase II (CaMKII), protein kinase C epsilon (PKCε), and mitogen activated protein kinase kinase (MEK). Moreover, schisandrin induced calcium influx, and phosphorylation of CaMKII, PKCε, and MEK. Inhibition of CAMKII and PKCε attenuated the schisandrin-induced phosphorylation of PKCε and MEK, and the phosphorylation of MEK, respectively. Moreover, schisandrin also stimulated the phosphorylation of cyclic AMP responsive-element binding protein (CREB) at Ser-133, an effect that was blocked by KN93. In addition to its neuritogenic effects, schisandrin increased the numbers of postsynaptic density-95-positive and FM1-43-positive puncta in dendrites and synaptic boutons, respectively. CONCLUSION: In hippocampal neurons, schisandrin exhibits neurotrophic properties that are mediated by the CaMKII-PKCε-MEK pathway.

A multi-D-shaped optical fiber for refractive index sensing.

A novel class of multi-D-shaped optical fiber suited for refractive index measurements is presented. The multi-D-shaped optical fiber was constructed by forming several D-sections in a multimode optical fiber at localized regions with femtosecond laser pulses. The total number of D-shaped zones fabricated could range from three to seven. Each D-shaped zone covered a sensor volume of 100 µm depth, 250 µm width, and 1 mm length. The mean roughness of the core surface obtained by the AFM images was 231.7 nm, which is relatively smooth. Results of the tensile test indicated that the fibers have sufficient mechanical strength to resist damage from further processing. The multi-D-shaped optical fiber as a high sensitive refractive-index sensor to detect changes in the surrounding refractive index was studied. The results for different concentrations of sucrose solution show that a resolution of 1.27 × 10(-3)-3.13 × 10(-4) RIU is achieved for refractive indices in the range of 1.333 to 1.403, suggesting that the multi-D-shaped fibers are attractive for chemical, biological, and biochemical sensing with aqueous solutions.

Role of PKA in the anti-Thy-1 antibody-induced neurite outgrowth of dorsal root ganglionic neurons.

Thy-1 is highly expressed in the mammalian nervous system. Our previous study showed that addition of anti-Thy-1 antibody to cultured dorsal root ganglionic (DRG) neurons promotes neurite outgrowth. In this study, we identified a novel signaling pathway mediating this event. Treatment with function-blocking anti-Thy-1 antibodies enhanced neurite outgrowth of DRG neurons in terms of total neurite length, longest neurite length, and total neurite branching points. To elucidate the possible signal transduction pathway involved, activation of kinases was evaluated by Western blotting. Transient phosphorylation of protein kinase A (PKA) and mitogen-activated kinase kinase (MEK) was induced after 15 min of anti-Thy-1 antibody treatment. Pretreatment with a PKA inhibitor (PKI) or an MEK inhibitor, PD98059, significantly decreased the neurite outgrowth response triggered by anti-Thy-1 antibody, indicating the involvement of both kinases. In addition, anti-Thy-1 antibody treatment also induced transient phosphorylation of cyclic AMP-response element-binding protein (CREB) and this effect was also blocked by a PKI or PD98059. Furthermore, the fact that PKI abolished anti-Thy-1 antibody-induced MEK phosphorylation showed that PKA acts upstream of the MEK-CREB cascade. In summary, the PKA-MEK-CREB pathway is a new pathway involved in the neurite outgrowth-promoting effect of anti-Thy-1 antibody.

Multiple negative feedbacks on CD152 expression in allograft tolerance.

BACKGROUND: CD152 has been implicated in tolerance induction. This study investigated how CD80 and CD86 regulated CD152 expression in a low-responding cardiac transplant model with CD152-mediated long-term graft acceptance. METHODS: A low-responding cardiac transplant model from BALB/c to B10.A was used. Donor-specific stimulation and multiple antibody blockade of the CD80/CD86:CD28/CD152 co-stimulatory pathway was applied to the splenic T cells from B10.A recipients with 100-day grafts (B10.A-100). Proliferation assays, quantitative (Q) real-time polymerase chain reaction (PCR), flow cytometric analyses, and fluorescence microscopy were conducted to examine the roles of CD80 and CD86 in CD152 expression. RESULTS: B10.A-100 splenic T cells were hyporesponsive to donor-specific stimulation, and anti-CD80, anti-CD86, or anti-CD152 treatment significantly enhanced the proliferation response of the B10.A-100 splenic T cells. Proliferation assays and Q-PCR revealed that CD152 inhibited T-cell proliferation and, at the same time, decreased CD152 expression by secluding CD80 and CD86 from CD28 engagement. Flow cytometric analyses and fluorescence microscopy showed that CD28 engagement facilitated intracellular accumulation of CD152. Besides, CD152 engagement by CD80 decreased CD152 mRNA transcription, and CD152 engagement by CD86 inhibited surface expression of CD152. CONCLUSIONS: CD80 and CD86 controlled CD152-mediated allograft tolerance by multiple negative feedbacks on CD152 mRNA and surface expression.

Role of Thy-1 in in vivo and in vitro neural development and regeneration of dorsal root ganglionic neurons.

We have examined the expression of Thy-1, an abundant glycosylphosphatidylinositol (GPI)-anchored glycoprotein, in dorsal root ganglia (DRG) and associated nerve fascicles, during postnatal development and following a nerve crush. The expression levels of Thy-1 in DRG neurons, dorsal roots, and central processes in spinal cord were rather low at postnatal day 2, and gradually increased as DRG neurons matured. During early development, the expression of Thy-1 within DRG neurons was low and equally distributed between plasma membrane and cytosol. With maturation, the staining intensities of Thy-1 in both the plasma membrane and the cytosol of DRG neurons became increased. We also studied Thy-1 expression in the regeneration of mature DRG neurons following the crush injury of sciatic nerve. Two days after the crush injury, Thy-1 expression dramatically decreased in the DRG neurons on the lesion side. Between 4 and 7 days after the injury, the expression of Thy-1 gradually increased and returned to a normal level 1 week after the sciatic nerve crush. The time course of the up-regulation of Thy-1 expression during regeneration matched that of the recovery of sensory functions, such as pain withdraw reflex, placing reflex, and the score of Basso-Beattie-Bresnahan Locomotor Rating Scale. Taken together, our results suggest that Thy-1 expression is developmentally regulated and is closely associated with the functional maturation of DRG neurons during both postnatal development and nerve regeneration. Furthermore, perturbation of Thy-1 function with anti-Thy-1 antibodies promoted neurite outgrowth from primary cultured DRG neurons, again confirming the inhibitory role of Thy-1 on neurite outgrowth.

Dual behavior of masked o-benzoquinones in intermolecular Diels-Alder reactions with acyclic dienes: a rapid entry to polyfunctionalized bicyclo[2.2.2]oct-5-en-2-ones and cis-decalins.

The potentiality of the masked o-benzoquinones, i.e., 6,6-dimethoxy-2,4-cyclohexadienones 5-8, to react both as dienes and dienophiles in their intermolecular reactions has been demonstrated. The masked o-benzoquinones (MOBs) 5-8 generated in situ from 2-methoxyphenols 1-4 underwent intermolecular Diels-Alder cycloadditions with acyclic 1,3-dienes 9a-e to provide bicyclo[2.2.2]octenones 10a-f-13a-f along with cis-decalin derivatives 14a-f-17a-f with regio- and stereoselectivity, except in the case of MOB 8. The formation of cis-decalins in these Diels-Alder reactions illustrates the dienophilic character of MOBs, in addition to their general behavior as dienes. The ratio of the two cycloadducts obtained in each reaction as a result of the dual character of MOBs depends on the nature and/or position of the substituents on both the cyclohexadienone moiety and the added conjugated acyclic diene. All of the cycloadducts resulted from the diene property of MOBs in intermolecular Diels-Alder reactions smoothly underwent Cope rearrangement to furnish cis-decalins as sole products in excellent to quantitative yields.

Applied anatomy of the genital branch of the genitofemoral nerve in open inguinal herniorrhaphy.

OBJECTIVE: To record the anatomical variations and clinical importance of the genital branch of the genitofemoral nerve in the inguinal canal. DESIGN: Neuropathology study. SETTING: General hospital, Taiwan. SUBJECTS: 58 cadavers. INTERVENTIONS: 116 dissections findings of the genitofemoral nerve in the inguinal canal. MAIN OUTCOME MEASURES: Anatomical variations. RESULTS: All the genital branches passed through the ventral aspect of the internal ring. Almost all these branches entered the ring and continued within the spermatic cord between the fibres of the cremaster and the internal spermatic fascia. In 59% the nerve was related to the inferior fibres and in 38% it was in relation to the lateral or medial fibres. In only 3% did the genital branch run outside the spermatic cord. CONCLUSION: To avoid damage to the genital branch of the genitofemoral nerve, suturing should be done over the dorsal aspect of the internal ring. The cremaster should be incised longitudinal rather than transversely and the genital branch of the genitofemoral nerve should be identified before repair of the posterior wall.

Dual SPECT of dopamine system using [99mTc]TRODAT-1 and [123I]IBZM in normal and 6-OHDA-lesioned formosan rock monkeys.

Parkinson's disease (PD) is a progressive neurodegenerative disorder characterized by a severe loss of the dopaminergic neurons in the substantia nigra pars compacta. In this study, we evaluated pre- and post-synaptic binding sites of the dopamine system in three normal and one parkinsonian monkeys using simultaneous [99mTc]TRODAT-1 and [123I]IBZM imaging. The parkinsonian monkey was induced by injecting 6-hydroxydopamine (6-OHDA) bilaterally into the medial forebrain bundle under MRI guidance. [99mTc]TRODAT-1 (targeting dopamine transporters) and [123I]IBZM (targeting D(2)/D(3) receptors) were administered almost simultaneously and the SPECT images were acquired over 4 h using a dual-headed gamma camera equipped with ultra-high resolution fan-beam collimators. Data were obtained using energy window of 15% centered on 140 keV for 99mTc in conjunction with 10% asymmetric energy window in a lower bound at 159 keV for 123I. Single SPECT studies of [99mTc]TRODAT-1 and [123I]IBZM were also performed. We found a comparable image quality and uptake ratios between single- and dual-isotope studies. There are higher TRODAT-1 uptakes in the control monkeys than the 6-OHDA-lesioned monkey. The uptake of [123I] IBZM showed no significant difference between controls and 6-OHDA-lesioned monkey. Our results suggest that dual isotope imaging using [99mTc]TRODAT-1 and [123I]IBZM may be a useful means in evaluating the changes of both pre- and post-synaptic dopamine system in a primate model of parkinsonism.