Injection Buffer Research Articles

Injection Buffer for Mosquitoes

Injection Buffer for Mosquitoes

Hyperpolarization of 15N‐Pyridinium by Using Parahydrogen Enables Access to Reactive Oxygen Sensors and Pilot In Vivo Studies

AbstractMagnetic resonance with hyperpolarized contrast agents is one of the most powerful and noninvasive imaging platforms capable for investigating in vivo metabolism. While most of the utilized hyperpolarized agents are based on 13C nuclei, a milestone advance in this area is the emergence of 15N hyperpolarized contrast agents. Currently, the reported 15N hyperpolarized agents mainly utilize the dissolution dynamic nuclear polarization (d‐DNP) protocol. The parahydrogen enhanced 15N probes have proven to be elusive and have been tested almost exclusively in organic solvents. Herein, we designed a reaction based reactive oxygen sensor 15N‐boronobenzyl‐2‐styrylpyridinium (15N‐BBSP) which can be hyperpolarized with para‐hydrogen. Reactive oxygen species plays a vital role as one of the essential intracellular signalling molecules. Disturbance of the H2O2 level usually represents a hallmark of pathophysiological conditions. This H2O2 probe exhibited rapid responsiveness toward H2O2 and offered spectrally resolvable chemical shifts. We also provide strategies to bring the newly developed probe from the organic reaction solution into a biocompatible injection buffer and demonstrate the feasibility of in vivo 15N signal detection. The present work manifests its great potential not only for reaction based reactive sensing probes but also promises to serve as a platform to develop other contrast agents.

Hyperpolarization of 15N-Pyridinium by Using Parahydrogen Enables Access to Reactive Oxygen Sensors and Pilot In Vivo Studies.

Magnetic resonance with hyperpolarized contrast agents is one of the most powerful and noninvasive imaging platforms capable for investigating in vivo metabolism. While most of the utilized hyperpolarized agents are based on 13C nuclei, a milestone advance in this area is the emergence of 15N hyperpolarized contrast agents. Currently, the reported 15N hyperpolarized agents mainly utilize the dissolution dynamic nuclear polarization (d-DNP) protocol. The parahydrogen enhanced 15N probes have proven to be elusive and have been tested almost exclusively in organic solvents. Herein, we designed a reaction based reactive oxygen sensor 15N-boronobenzyl-2-styrylpyridinium (15N-BBSP) which can be hyperpolarized with para-hydrogen. Reactive oxygen species plays a vital role as one of the essential intracellular signalling molecules. Disturbance of the H2O2 level usually represents a hallmark of pathophysiological conditions. This H2O2 probe exhibited rapid responsiveness toward H2O2 and offered spectrally resolvable chemical shifts. We also provide strategies to bring the newly developed probe from the organic reaction solution into a biocompatible injection buffer and demonstrate the feasibility of in vivo 15N signal detection. The present work manifests its great potential not only for reaction based reactive sensing probes but also promises to serve as a platform to develop other contrast agents.

N-buffer design and optimization in 4500V class IGBT

In this paper, the effects of doping structure of multi-proton injection (MPI) buffer layer on static and short circuit characteristics of IGBT were studied, and the relationship between donor concentration of the buffer layer and injection dose and annealing temperature was studied. The MPI buffer layer suitable for 4500V IGBT was optimized by comparative experiments. This MPI buffer layer can solve the problem of high-temperature activation of the traditional IGBT buffer layer and lower thermal budget and can be fabricated on large wafers for better parameter consistency.

A Continuous K-Band Phase Shifter Based on Injection-Locked Dual Voltage-Controlled Oscillators

In this letter, a <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"> <tex-math notation="LaTeX">$K$ </tex-math></inline-formula> -band signal generation phase shifter (SGPS) is implemented using a standard 55-nm complementary metal-oxide semiconductor (CMOS). Its phase shifting is achieved by injecting signal of a voltage-controlled oscillator (VCO) to the other through an injection buffer. Besides continuous phase shift, it can offer a signal source, facilitating multi-phase signal generation in phase array transceivers. Measurement results suggest the prototype implemented in 55-nm CMOS is able to provide a frequency tuning range of 22.5 to 24.8 GHz and minimal output power of 5.2 dBm. It demonstrates a maximal tuning phase of 45.5°.

Separation of unsaturated C18 fatty acids using perfluorinated-micellar electrokinetic chromatography: II) role of nano-structuration

• Free fatty acids are separated by capillary electrophoresis using fluorosurfactants. • Scattering experiments evidence aggregation from small micelles to large structures. • Micelle structuration in injection buffer and background electrolytes is affected by fatty acids. • Selective interactions between fatty acids and fluorosurfactant allow CE separation. Three fatty acids were separated by micellar electrokinetic chromatography (MEKC) using ammonium perfluorooctanoate as volatile surfactant. Characterizations of both injection buffer and background electrolyte have been performed by complementary techniques including scattering experiments (light, x-rays) and NMR. Different structures have been observed such as small micelles at low methanol content and larger aggregates at high methanol content. In both cases fatty acids interact specifically with the perfluorinated self-assemblies due to different geometrical constraints and different interactions between polar head groups, ensuring separation by MEKC.

Application of monitoring method in the establishment and maintenance of gas cap injection barrier in a Oilfield

In order to make technical reserves for developing gas cap and buffer zone in A Oilfield and explore effective development methods of gas cap reservoir, according to the overall development idea of oil-gas ring, a field test of gas cap injection and polymer barrier development buffer zone in Layer B was carried out in the structural axis of South Block in 2006. The test adopted the development mode of gas cap injection and polymer barrier development buffer zone, that is, the barrier was established by injecting polymer solution at the oil-gas boundary to separate natural gas from crude oil in buffer zone and keep relatively stable. In this paper, the morphological changes during the establishment and maintenance of barriers are found out by means of monitoring.

Site-Specific Radiolabeling of a Human PD-L1 Nanobody via Maleimide-Cysteine Chemistry.

Immune checkpoint inhibitors targeting the programmed cell death-1 (PD-1) and its ligand PD-L1 have proven to be efficient cancer therapies in a subset of patients. From all the patients with various cancer types, only 20% have a positive response. Being able to distinguish patients that do express PD-1/PD-L1 from patients that do not allows patients to benefit from a more personalized and efficient treatment of tumor lesion(s). Expression of PD-1 and PD-L1 is typically assessed via immunohistochemical detection in a tumor biopsy. However, this method does not take in account the expression heterogeneity within the lesion, nor the possible metastasis. To visualize whole-body PD-L1 expression by PET imaging, we developed a nanobody-based radio-immunotracer targeting PD-L1 site-specifically labeled with gallium-68. The cysteine-tagged nanobody was site-specifically conjugated with a maleimide (mal)-NOTA chelator and radiolabeling was tested at different nanobody concentrations and temperatures. Affinity and specificity of the tracer, referred to as [68Ga]Ga-NOTA-mal-hPD-L1 Nb, were assayed by surface plasmon resonance and on PD-L1POS or PD-L1NEG 624-MEL cells. Xenografted athymic nude mice bearing 624-MEL PD-L1POS or PD-L1NEG tumors were injected with the tracer and ex vivo biodistribution was performed 1 h 20 min post-injection. Ideal 68Ga-labeling conditions were found at 50 °C for 15 min. [68Ga]Ga-NOTA-mal-hPD-L1 Nb was obtained in 80 ± 5% DC-RCY with a RCP > 99%, and was stable in injection buffer and human serum up to 3 h (>99% RCP). The in vitro characterization showed that the NOTA-functionalized Nb retained its affinity and specificity. Ex vivo biodistribution revealed a tracer uptake of 1.86 ± 0.67% IA/g in the positive tumors compared with 0.42 ± 0.04% IA/g in the negative tumors. Low background uptake was measured in the other organs and tissues, except for the kidneys and bladder, due to the expected excretion route of Nbs. The data obtained show that the site-specific 68Ga-labeled NOTA-mal-hPD-L1 Nb is a promising PET radio-immunotracer due to its ease of production, stability and specificity for PD-L1.

ALPHA: A Learning-Enabled High-Performance Network-on-Chip Router Design for Heterogeneous Manycore Architectures

Heterogeneous manycores comprised of CPUs, GPUs and accelerators are putting stringent demands on network-on-chips (NoCs). The NoCs need to support the combined traffic, including both latency-sensitive CPU traffic and throughput-sensitive GPU and accelerator traffic. We study the characteristics of the combined traffic, and observe that (1) the limited injection bandwidth is the main obstacle to throughput improvement, and (2) the latency due to local and global contention accounts for a significant portion of the network latency. We propose a router architecture named ALPHA for heterogeneous manycores. ALPHA introduces two new optimizations: (1) increasing injection bandwidth to improve throughput, and (2) resolving local and global contention to reduce network latency. Specifically, ALPHA increases the injection bandwidth through modifications to injection link, crossbar switch and buffer organization in the injection port of the router; ALPHA identifies the upcoming local contention and resolves it by optimally selecting traffic routes; ALPHA detects and alleviates the global contention by utilizing a supervised learning engine for traffic analysis, prediction, and adjustment. Simulation results using Rodinia benchmark show that ALPHA provides 28 percent throughput increase, 24 percent latency reduction, 22 percent execution time speedup, and 19 percent energy efficiency improvement, compared to the baseline router.

Impact of injection buffer volume to perform bronchoalveolar lavage fluid collection for isolating alveolar macrophages to investigate fine particle-induced IL-1α secretion

The importance of alveolar macrophages has been reported in many toxicology/immunology studies. Alveolar macrophages release interleukin (IL)-1α as a damage-associated molecular pattern (DAMP) when stimulated by fine particles. However, it is unclear whether cell isolation procedures affect ex vivo particle-induced responses in primary mouse alveolar macrophages (mAM). In this study, effects of injection buffer volume used to perform bronchoalveolar lavage fluid (BALF) collection to isolate mAM for use in ex vivo particle-induced responses were assessed. Among the mAM obtained from BALF collected using a 0.55 or 0.75 ml, but not a 1.0 ml buffer injection volume, decreased cell viability and IL-1α release were observed when cells were stimulated ex vivo with silica crystal or aluminum salt. Injected buffer composition did not affect the IL-1α release. On the other hand, IL-6 secretion induced by lipopolysaccharide (LPS) did not differ among mAM obtained from BALF collected using the different volumes. Expression levels of cell surface markers like CD11c, SiglecF, and CD64 did not differ among mAM obtained from BALF collected using the different injection buffer volumes. IL-1α release (and also necroptosis) induced by ex vivoparticle stimulation was suppressed by RIPK3 inhibitor or cytochalasin D co-treatment. Decreases in RIPK3 phosphorylation were noted in mAM obtained in BALF collected using the 1.0 ml injection volume compared with mAM obtained in BALF using 0.55 or 0.75 ml buffer. These observations illustrate that larger volumes of buffer used to collect BALF from mice can affect sensitivity of the isolated mAM to ex vivo particle-induced responses by inhibiting their functions.

Performance analysis of network-on-chip in many-core processors

Network-on-chip (NoC) is an integral part of many-core microprocessors. Performance analysis of network-on-chip directly affects the performance of the microprocessor. In this paper we propose a mathematical model to represent packet flow in an NoC as an open feed-forward queuing network. We study the performance of NoC by varying different parameters that includes packet injection rate, packet size, buffer size and number of virtual channels. We also discuss how different flow control algorithms, injection processes, traffic patterns can be incorporated in our model. Apart from the speedup achieved by our model, we also demonstrate that our model can be used to explore various configurations of NoC with minimal error.

Security vulnerabilities related to web-based data

In this digital era, organizations and industries are moving towards replacing websites with web applications for many obvious reasons. With this transition towards web-based applications, organizations and industries find themselves surrounded by several threats and vulnerabilities. One of the largest concerns is keeping their infrastructure safe from attacks and misuse. Web security entails applying a set of procedures and practices, by applying several security principles at various layers to protect web servers, web users, and their surrounding environment. In this paper, we will discuss several attacks that may affect web-based applications namely: SQL injection attacks, cookie poisoning, cross-site scripting, and buffer overflow. Additionally, we will discuss detection and prevention methods from such attacks.

Enhanced charge injection in 6, 13-bis(triisopropylsilylethylnyl)-pentacene field-effect transistors with a rhenium oxide buffer layer

We introduce a transition metal oxide, rhenium trioxide (ReO3), as a charge injection buffer layer for 6,13-bis(triisopropylsilylethylnyl)-pentacene (TIPS-pentacene) field-effect transistors (FETs). By inserting a ReO3 layer, a large energy barrier between silver source electrode and TIPS-pentacene layer was significantly reduced. While the TIPS-pentacene FETs showed low hole mobility (μh) of 0.25 cm2 V−1 s−1 and large threshold voltage (VTH) of −25 V due to large contact resistance (RC) of 155 kΩ cm, the TIPs-pentacene FETs with ReO3 decreased RC as low as 28 kΩ cm. Thus, we can improve μh to two times greater (∼0.46 cm2 V−1 s−1) and reduce VTH to around 0 V. Together with these improved electrical characteristics, the TIPS-pentacene with ReO3 shows stable operation under gate voltage bias stress conditions. The shift of VTH and degradation of μh which were shown in the TIPS-pentacene FET without ReO3 is suppressed in the TIPS-pentacene FETs with ReO3 under both positive and negative bias conditions.

Injection Buffer for Xenopus

Injection Buffer for <i>Xenopus</i>

CRISPR/Cas9 mediated high efficiency knockout of the eye color gene Vermillion in Helicoverpa zea (Boddie).

Among various genome editing tools available for functional genomic studies, reagents based on clustered regularly interspersed palindromic repeats (CRISPR) have gained popularity due to ease and versatility. CRISPR reagents consist of ribonucleoprotein (RNP) complexes formed by combining guide RNA (gRNA) that target specific genomics regions and a CRISPR associated nuclease (Cas). The gRNA targeting specific gene sequences may be delivered as a plasmid construct that needs to be transcribed or as a synthetic RNA. The Cas nuclease can be introduced as a plasmid construct, mRNA, or purified protein. The efficiency of target editing is dependent on intrinsic factors specific to each species, the target gene sequence, and the delivery methods of CRISPR gRNA and the Cas nuclease. Although intrinsic factors affecting genome editing may not be altered in most experiments, the delivery method for CRISPR/Cas reagents can be optimized to produce the best results. In this study, the efficiency of genome editing by CRISPR/Cas system in the bollworm, Helicoverpa zea (Boddie), was evaluated using ribonucleoprotein (RNP) complexes assembled by binding synthetic gRNA with purified Cas9 nuclease engineered with nuclear localization signals to target the vermillion (eye color) gene. Mutation rates of adults emerging from embryos microinjected with 1, 2, or 4 μM RNP complexes were compared using replicated experiments. Embryos injected with 2 or 4 μM RNP complexes displayed significantly higher mutation rates (>88%) in surviving adults compared to those injected with 1 μM. The hatch rate in embryos injected with RNP complexes and with injection buffer only (mock injections) was reduced by 19.8(±5.2)% compared to noninjected control embryos, but did not differ significantly between injected embryos. Evaluation of potential off-target sites in H. zea genome did not identify any mutations. This study demonstrates that in vitro assembled synthetic RNP complexes can be used to obtain high genome editing rates in a reproducible manner in functional genomics or genetic manipulation studies.

Impact of Subolesin and Cystatin Knockdown by RNA Interference in Adult Female Haemaphysalis longicornis (Acari: Ixodidae) on Blood Engorgement and Reproduction.

Currently, multi-antigenic vaccine use is the method of choice for the strategic control of ticks. Therefore, determining the efficacy of combined antigens is a promising avenue of research in the development of anti-tick vaccines. The antigen responsible for blood intake and reproduction has proven suitable as a vaccine antigen. It has been shown to silence Haemaphysalis longicornis salivary cystatin (HlSC-1) and subolesin by RNA interference. Adult unfed female ticks were injected with double-stranded RNA of (A) subolesin, (B) cystatin, (C) subolesin plus cystatin, and (D) injection buffer, then fed alongside normal unfed males up to spontaneous drop-down. The percentage of knockdowns was determined by real-time polymerase chain reaction. Sixty-three percent and 53% knockdown rates were observed in subolesin and cystatin double-stranded RNA-injected ticks respectively, while 32 and 26% knockdown rates of subolesin and cystatin transcript were observed in subolesin plus cystatin double-stranded RNA-injected ticks. Subolesin and/or cystatin knockdown causes a significant (p < 0.05) reduction in tick engorgement, egg mass weight, and egg conversion ratio. Most importantly, combined silencing did not act synergistically, but caused a similarly significant (p < 0.05) reduction in tick engorgement, egg mass weight, and egg conversion ratio. Therefore, the elucidation of multiple antigens may be helpful in the future of vaccines.

Energy Level Alignment at the Fullerene/Titanium Oxide Ultrathin Film Interface

The performance of molecule based electronic devices can be improved by use of transition metal oxides (TMO) as charge injection buffer layers between electrodes and organic semiconductors. It is known that the appropriate energy level alignment at the molecule/TMO interface determines the efficiency of the respective TMO. Herein, scanning tunneling microscopy is employed to characterize the interface formed by fullerenes (C60) deposited on a titanium oxide (TiO) ultrathin film, which is created by oxidation of a Pt3Ti(111) surface. Individual C60 are identified with orbital resolution, and the interfacial and intermolecular interactions are characterized in detail. Furthermore, the energy level alignment at the C60/TiO ultrathin film interface is deduced based on scanning tunneling spectroscopy data. The results demonstrate that the C60/TiO interface corresponds to a type-I heterojunction and, thus, is useful to decouple C60 molecules from the metallic alloy surface.

Transition metal oxide as anode interface buffer for impedance spectroscopy

Impedance spectroscopy is a strong method in electric measurement, which also shows powerful function in research of carrier dynamics in organic semiconductors when suitable mathematical physical models are used. Apart from this, another requirement is that the contact interface between the electrode and materials should at least be quasi-ohmic contact. So in this report, three different transitional metal oxides, V2O5, MoO3 and WO3 were used as hole injection buffer for interface of ITO/NPB. Through the impedance spectroscopy and PSO algorithm, the carrier mobilities and I-V characteristics of the NPB in different devices were measured. Then the data curves were compared with the single layer device without the interface layer in order to investigate the influence of transitional metal oxides on the carrier mobility. The careful research showed that when the work function (WF) of the buffer material was just between the work function of anode and the HOMO of the organic material, such interface material could work as a good bridge for carrier injection. Under such condition, the carrier mobility measured through impedance spectroscopy should be close to the intrinsic value. Considering that the HOMO (or LUMO) of most organic semiconductors did not match with the work function of the electrode, this report also provides a method for wide application of impedance spectroscopy to the research of carrier dynamics.

Vitellogenin-RNAi and ovariectomy each increase lifespan, increase protein storage, and decrease feeding, but are not additive in grasshoppers.

Reduced reproduction has been shown to increase lifespan in many animals, yet the mechanisms behind this trade-off are unclear. We addressed this question by combining two distinct, direct means of life-extension via reduced reproduction, to test whether they were additive. In the lubber grasshopper, Romalea microptera, ovariectomized (OVX) individuals had a ~20% increase in lifespan and a doubling of storage relative to controls (Sham operated). Similarly, young female grasshoppers treated with RNAi against vitellogenin (the precursor to egg yolk protein) had increased fat body mass and halted ovarian growth. In this study, we compared VgRNAi to two control groups that do not reduce reproduction, namely buffer injection (Buffer) and injection with RNAi against a hexameric storage protein (Hex90RNAi). Each injection treatment was tested with and without ovariectomy. Hence, we tested feeding, storage, and lifespans in six groups: OVX and Buffer, OVX and Hex90RNAi, OVX and VgRNAi, Sham and Buffer, Sham and Hex90RNAi, and Sham and VgRNAi. Ovariectomized grasshoppers and VgRNAi grasshoppers each had similar reductions in feeding (~40%), increases in protein storage in the hemolymph (150-300%), and extensions in lifespan (13-21%). Ovariectomized grasshoppers had higher vitellogenin protein levels than did VgRNAi grasshoppers. Last but not least, when ovariectomy and VgRNAi were applied together, there was no greater effect on feeding, protein storage, or longevity. Hence, feeding regulation, and protein storage in insects, may be conserved components of life-extension via reduced reproduction.

The effect of metal-buffer bilayer drain/source electrodes on the operational stability of the organic field effect transistors

In this paper, we have investigated experimentally the effect of different drain/source (D/S) electrodes and charge injection buffer layers on the electrical properties and operational stability of a stilbene organic field effect transistor (OFET). The results show that the organic buffer layer of copper phthalocyanine (CuPc) considerably improves the electrical properties of the transistors, but has a negligible effect on their temporal behavior. On the other hand, inorganic metal-oxide buffer layer of molybdenum oxide (MoO3) drastically changes both the electrical properties and operational stability. The functionalities of this metal-oxide tightly depend on the properties of the D/S metallic electrodes. OFETs with Al/MoO3 as the bilayer D/S electrodes have the best electrical properties: field effect mobility μeff=0.32cm2V−1s−1 and threshold voltage VTH=−5V and the transistors with Ag/MoO3 have the longest operational stability. It was concluded that the chemical stability of the metal/metal-oxide or metal/organic interfaces of the bilayer D/S electrodes determine the operational stability of the OFETs.