A simple method for gene expression in endo- and ectodermal cells in mouse embryos before neural tube closure.

The lack of a widely accessible method for expressing genes of interest in wild-type embryos is a fundamental obstacle to understanding genetic regulation during embryonic development. In particular, only a few methods are available for introducing gene expression vectors into cells prior to neural tube closure, which is a period of drastic development for many tissues. In this study, we present a simple technique for injecting vectors into the amniotic cavity and allowing them to reach the ectodermal cells and the epithelia of endodermal organs of mouse embryos at E8.0 via in utero injection, using only a widely used optical fiber with an illuminator. Using this technique, retroviruses can be introduced to facilitate the labeling of cells in various tissues, including the brain, spinal cord, epidermis, and digestive and respiratory organs. We also demonstrated in utero electroporation of plasmid DNA into E7.0 and E8.0 embryos. Taking advantage of this method, we reveal the association between Ldb1 and the activity of the Neurog2 transcription factor in the mouse neocortex. This technique can aid in analyzing the roles of genes of interest during endo- and ectodermal development prior to neural tube closure.

Analyzing the Automatic Power Level Control Effect of a Signal Generator in RF Power Sensor Calibration by a Direct Comparison Transfer Method and a Millimeter Wave Application.

Most calibration laboratories prefer the Direct Comparison Transfer Method (DCTM) for a reliable and accurate calibration of power sensors in the radio frequency (RF) scope. Most studies suggest using this calibration method, with its automatic power level control (APLC) of RF signal generators. The APLC is preferred to keep the output power level of the signal generator the same, while the power sensor is calibrated and the reference power sensor is connected to the measurement system. The known APLC mechanisms are also explained for the DCTM, and a comparison of the calibration factor values carried out with and without the automatic power level control process in the DCTM is also given in this study. RF power sensor calibrations with coaxial and waveguide connector types are examined with DCTM in this study as well. This study shows that the DCTM, unless with APLC, should be applied for the waveguide power sensor's calibration at millimeter wave frequencies.

Land use and land cover dynamics in the Upper Ganga Riverine Wetland: unraveling ecosystem services over two decades.

Anthropogenic activities have drastically transformed natural landscapes, profoundly impacting land use and land cover (LULC) and, consequently, the provision and functionality of ecosystem service values (ESVs). Evaluating the changes in LULC and their influence on ESVs is imperative to protect ecologically fragile ecosystems from degradation. This study focuses on a highly sensitive Upper Ganga riverine wetland in India, covering Hapur, Amroha, Bulandshahr, and Sambhal districts, which is well-known for its significant endemic flora and fauna. The study analyzes the subtle variability in ecosystem services offered by the various LULC biomes, including riverine wetland, built-up, cropland, forest, sandbar, and unused land. LULC classification is carried out using Landsat satellite imagery 5 and 8 for the years 2000, 2010, and 2020, using the random forest method. The spatiotemporal changing pattern of ESVs is assessed utilizing the value transfer method with two distinct value coefficients: global value coefficients (C14) for a worldwide perspective and modified local value coefficients X08 for a more specific local context. The results show a significant increase in built-up and unused land, with a corresponding decrease in wetlands and forests from 2000 to 2020. The combined ESVs for all the districts are worth US $5072 million (C14) and US $2139 million (X08) in the year 2000, which declined to US $4510 million (C14) and US $1770 million (X08) in the year 2020. The sensitivity analysis reveals that the coefficient of sensitivity (CS) is below one for all biomes, suggesting the robustness of the employed value coefficients in estimating ESVs. Moreover, the analysis identifies cropland, followed by forests and wetlands, as the LULC biomes most responsive to changes. This research provides crucial insights to stakeholders and policymakers for developing sustainable land management practices aimed at enhancing the ecological worth of the Upper Ganga Riverine Wetland.

Large-scale and stacked transfer of bilayers MoS2devices on a flexible polyimide substrate.

Two-dimensional transition metal dichalcogenides (TMDs), as flexible and stretchable materials, have attracted considerable attention in the field of novel flexible electronics due to their excellent mechanical, optical, and electronic properties. Among the various TMD materials, atomically thin MoS2has become the most widely used material due to its advantageous properties, such as its adjustable bandgap, excellent performance, and ease of preparation. In this work, we demonstrated the practicality of a stacked wafer-scale two-layer MoS2film obtained by transferring multiple single-layer films grown using chemical vapor deposition. The MoS2field-effect transistor cell had a top-gated device structure with a (PI) film as the substrate, which exhibited a high on/off ratio (108), large average mobility (∼8.56 cm2V-1s-1), and exceptional uniformity. Furthermore, a range of flexible integrated logic devices, including inverters, NOR gates, and NAND gates, were successfully implemented via traditional lithography. These results highlight the immense potential of TMD materials, particularly MoS2, in enabling advanced flexible electronic and optoelectronic devices, which pave the way for transformative applications in future-generation electronics.

Electrochemical and thermal detection of allergenic substance lysozyme with molecularly imprinted nanoparticles.

Lysozyme (LYZ) is a small cationic protein which is widely used for medical treatment and in the food industry to act as an anti-bacterial agent; however, it can trigger allergic reactions. In this study, high-affinity molecularly imprinted nanoparticles (nanoMIPs) were synthesized for LYZ using a solid-phase approach. The produced nanoMIPs were electrografted to screen-printed electrodes (SPEs), disposable electrodes with high commercial potential, to enable electrochemical and thermal sensing. Electrochemical impedance spectroscopy (EIS) facilitated fast measurement (5-10 min) and is able to determine trace levels of LYZ (pM) and can discriminate between LYZ and structurally similar proteins (bovine serum albumin, troponin-I). In tandem, thermal analysis was conducted with the heat transfer method (HTM), which is based on monitoring the heat transfer resistance at the solid-liquid interface of the functionalized SPE. HTM as detection technique guaranteed trace-level (fM) detection of LYZ but needed longer analysis time compared to EIS measurement (30 min vs 5-10 min). Considering the versatility of the nanoMIPs which can be adapted to virtually any target of interest, these low-cost point-of-care sensors hold great potential to improve food safety.

A non-destructive and efficient transfer method for preparing 2D materials samples for transmission electron microscopy study.

Studying two-dimensional (2D) materials using transmission electron microscopy (TEM) is necessary and very important in many aspects. However, some 2D materials are not resistant to acids or alkalis, which are widely used in normal wet transfer techniques to transfer the exfoliated 2D nanosheets onto the TEM grids. On the other hand, dry stamping method can damage the holey carbon film on the TEM grids. In this article, we present a non-destructive, efficient, and widely applicable transfer method for preparing the TEM samples of the exfoliated 2D materials. Our method only uses the heat-release tape, PMMA, and blue Nitto tape. Neither acid nor alkali is involved in our method, therefore, impurities and damage can be avoided to the greatest extent. The method is also very efficient and can be accomplished in less than 30 min after the exfoliation of the 2D materials. This method is particularly useful for preparing the TEM samples of the 2D materials that are not resistant to acids and alkalis. The present method is also applicable to various 2D materials and various substrates.

Life Cycle Assessment and Cost-Benefit Analysis of Technologies in Water Resource Recovery Facilities: The Case of Sludge Pyrolysis.

In Europe, sewage sludge is mostly used in agriculture (49%) or incinerated (25%). Technologies for sludge management that can support the transformation of wastewater treatment plants (WWTPs) to water resource recovery facilities (WRRFs) are emerging. Sludge pyrolysis is one of them. It can generate two main high-value co-products: heat and biochar. Moreover, biochar can be transformed into activated carbon. The economic and environmental impacts of sludge pyrolysis and its comparison to the direct application of sludge in agriculture and incineration are unknown. Therefore, we applied a life cycle assessment (LCA) and a cost-benefit analysis (CBA) of sludge pyrolysis. We quantified environmental externalities in an LCA and then applied the benefit transfer method to monetize these externalities, which were included in an economic CBA. Pyrolysis reduced impacts in five to nine LCA categories and had a positive economic net present value (NPV) compared to using sludge in agriculture. Pyrolysis with biochar production was not better than incineration, showing increased impacts in nine categories and negative NPVs (-19 to -22 €/t sludge). The factor driving differences between the alternatives was the assumed CO2 externality price (164 €/ton CO2-eq) and the removal rate of pharmaceutical micropollutants of the sludge-based activated carbon. High uncertainty in environmental prices is one of the limitations of our study.

Influence of design and material characteristics on 3D printed flow-cells for heat transfer-based analytical devices.

Redesigning 3D-printed flow cells is reported used for heat transfer based detection of biomolecules from a flow-through system to an addition-type measurement cell. The aim of this study is to assess the performance of this new measurement design and critically analyse the influence of material properties and 3D printing approach on thermal analysis. Particular attention is paid to reduce the time to stabilisation, the sample volume in order to make the technique suitable for clinical applications, and improving the sensitivity of the platform by decreasing the noise and interference of air bubbles. The three different approaches that were studied included a filament polylactic acid cell using only fused filament fabrication (FFF), a resin cell printed using stereolitography (SLA), and finally a design made of copper, which was manufactured by combining metal injection moulding (MIM) with fused filament fabrication (FFF). Computational fluid dynamic (CFD) modelling was undertaken using ANSYS Fluent V18.1 to provide insight into the flow of heat within the measurement cell, facilitating optimisation of the system and theoretical response speed.It was shown that the measurement cells using SLA had the lowest noise (~ 0.6%) and shortest measurement time (15 min), whereas measurement cells produced using other approaches had lower specificity or suffered from voiding issues. Finally, we assessed the potential of these new designs for detection of biomolecules and amoxicillin, a commonly used beta lactam antibiotic, to demonstrate the proof of concept. It can be concluded that the resin addition-type measurement cells produced with SLA are an interesting affordable alternative, which were able to detect amoxicillin with high sensitivity and have great promise for clinical applications due to the disposable nature of the measurement cells in addition to small sample volumes.

A solvent-compatible filter-transfer method of semi-transparent carbon-nanotube electrodes stacked with silver nanowires.

Low-density films of single-walled carbon nanotubes (SWNTs) can be used as a semi-transparent top electrode for all-solution-processed film devices; however, their semiconductor characteristics vary depending on the experimental factors in their dispersion into solvents, and the sublayers are damaged as a result of solvent incompatibility. In this study, we report a solvent-compatible filter-transfer method for SWNT films stacked with silver nanowires (AgNWs), and evaluate the semiconductor characteristics through the p/n heterojunction with a Si wafer (SWNT/Si). AgNWs and SWNTs were successively filtered through their aqueous dispersion solutions using a membrane filter. The stacked semi-transparent films (AgNW/SWNT films with controlled densities) were successfully transferred onto glass plates and Si wafers. The transmittance at 550 nm revealed a window between 60% and 80% with a narrow sheet resistance range between 11 and 23 Ω â–¡-1. The power conversion efficiency (PCE) of SWNT/Si was improved to 11.2% in a junction area of 0.031 cm2 through the use of spin-coated Nafion resins; however, the accumulated resistance of SWNTs drastically reduced the PCE to 2% as the area increased to ≥0.5 cm2. AgNWs maintained the PCE within a range of 10.7% to 8.6% for an area ranging from 0.031 cm2 to 1.13 cm2. All of the photovoltaic parameters were dependent on the junction areas, suggesting that AgNWs function as an effective current-collector layer on the semiconductor layer of SWNTs without direct contact of AgNWs with the Si surface. In addition, we report a solvent-compatible experiment for transferring AgNW/SWNT films onto a solvent-sensitive perovskite material (CH3NH3PbI3).

Surface Properties of CVD-Grown Graphene Transferred by Wet and Dry Transfer Processes.

Graphene, an atomically thin material, has unique electrical, mechanical, and optical properties that can enhance the performance of thin film-based flexible and transparent devices, including gas sensors. Graphene synthesized on a metallic catalyst must first be transferred onto a target substrate using wet or dry transfer processes; however, the graphene surface is susceptible to chemical modification and mechanical damage during the transfer. Defects on the graphene surface deteriorate its excellent intrinsic properties, thus reducing device performance. In this study, the surface properties of transferred graphene were investigated according to the transfer method (wet vs. dry) and characterized using atomic force microscopy, Raman spectroscopy, and contact angle measurements. After the wet transfer process, the surface properties of graphene exhibited tendencies similar to the poly(methyl methacrylate) residue remaining after solvent etching. The dry-transferred graphene revealed a surface closer to that of pristine graphene, regardless of substrates. These results provide insight into the utilization of wet and dry transfer processes for various graphene applications.

Modelling of Land Use/Cover and LST Variations by Using GIS and Remote Sensing: A Case Study of the Northern Pakhtunkhwa Mountainous Region, Pakistan.

Alteration in Land Use/Cover (LULC) considered a major challenge over the recent decades, as it plays an important role in diminishing biodiversity, altering the macro and microclimate. Therefore, the current study was designed to examine the past 30 years (1987−2017) changes in LULC and Land Surface Temperature (LST) and also simulated for next 30 years (2047). The LULC maps were developed based on maximum probability classification while the LST was retrieved from Landsat thermal bands and Radiative Transfer Equation (RTE) method for the respective years. Different approaches were used, such as Weighted Evidence (WE), Cellular Automata (CA) and regression prediction model for the year 2047. Resultantly, the LULC classification showed increasing trend in built-up and bare soil classes (13 km2 and 89 km2), and the decreasing trend in vegetation class (−144 km2) in the study area. In the next 30 years, the built-up and bare soil classes would further rise with same speed (25 km2 and 36.53 km2), and the vegetation class would further decline (−147 km2) until 2047. Similarly for LST, the temperature range for higher classes (27 -< 30 °C) increased by about 140 km2 during 1987−2017, which would further enlarge (409 km2) until 2047. The lower LST range (15 °C to <21 °C) showed a decreasing trend (−54.94 km2) and would further decline to (−20 km2) until 2047 if it remained at the same speed. Prospective findings will be helpful for land use planners, climatologists and other scientists in reducing the increasing LST associated with LULC changes.

Male-derived copulatory plugs enhance implantation success in female Mus musculus.

Among a wide diversity of sexually reproducing species, male ejaculates coagulate to form what has been termed a copulatory plug. A number of functions have been attributed to copulatory plugs, including the inhibition of female remating and the promotion of ejaculate movement. Here we demonstrate that copulatory plugs also influence the likelihood of implantation, which occurs roughly 4 days after copulation in mice. Using a bead transfer method to control for differences in ejaculate retention and fertilization rates, we show that implantation rates significantly drop among females mated to genetically engineered males incapable of forming plugs (because they lack functional transglutaminase 4, the main enzyme responsible for its formation). Surprisingly, this result does not correlate with differences in circulating progesterone levels among females, an important hormone involved in implantation. We discuss three models that connect male-derived copulatory plugs to implantation success, including the hypothesis that plugs contribute to a threshold amount of stimulation required for females to become receptive to implantation.

A General Wet Transferring Approach for Diffusion-Facilitated Space-Confined Grown Perovskite Single-Crystalline Optoelectronic Thin Films.

Hybrid perovskite single-crystalline thin films are promising for making high-performance perovskite optoelectronic devices due to their superior physical properties. However, it is still challenging to incorporate them into multilayer devices because of their on-substrate growth. Here, a wet transfer method is used in transferring perovskite single-crystalline films perfectly onto various target substrates. More importantly, large millimeter-scaled single-crystalline films can be obtained via a diffusion-facilitated space-confined growth method as thin as a few hundred nanometers, which are capable of sustaining excellent crystalline quality and morphology after the transferring process. The availability of these crystalline films offers us a convenient route to further investigate their intrinsic properties of hybrid perovskites. We also demonstrate that the wet transfer method can be used for scalable fabrication of perovskite single-crystalline film-based photodetectors exhibiting a remarkable photoresponsivity. It is expected that this transferring strategy would promise broad applications of perovskite single-crystalline films for more complex perovskite devices.

Color Image Generation from LiDAR Reflection Data by Using Selected Connection UNET.

In this paper, a modified encoder-decoder structured fully convolutional network (ED-FCN) is proposed to generate the camera-like color image from the light detection and ranging (LiDAR) reflection image. Previously, we showed the possibility to generate a color image from a heterogeneous source using the asymmetric ED-FCN. In addition, modified ED-FCNs, i.e., UNET and selected connection UNET (SC-UNET), have been successfully applied to the biomedical image segmentation and concealed-object detection for military purposes, respectively. In this paper, we apply the SC-UNET to generate a color image from a heterogeneous image. Various connections between encoder and decoder are analyzed. The LiDAR reflection image has only 5.28% valid values, i.e., its data are extremely sparse. The severe sparseness of the reflection image limits the generation performance when the UNET is applied directly to this heterogeneous image generation. In this paper, we present a methodology of network connection in SC-UNET that considers the sparseness of each level in the encoder network and the similarity between the same levels of encoder and decoder networks. The simulation results show that the proposed SC-UNET with the connection between encoder and decoder at two lowest levels yields improvements of 3.87 dB and 0.17 in peak signal-to-noise ratio and structural similarity, respectively, over the conventional asymmetric ED-FCN. The methodology presented in this paper would be a powerful tool for generating data from heterogeneous sources.

Single-Shot Detection of Neurotransmitters in Whole-Blood Samples by Means of the Heat-Transfer Method in Combination with Synthetic Receptors.

Serotonin is an important neurotransmitter that plays a major role in the pathogenesis of a variety of conditions, including psychiatric disorders. The detection of serotonin typically relies on high-performance liquid chromatography (HPLC), an expensive technique that requires sophisticated equipment and trained personnel, and is not suitable for point-of-care applications. In this contribution, we introduce a novel sensor platform that can measure spiked neurotransmitter concentrations in whole blood samples in a fast and low-cost manner by combining synthetic receptors with a thermal readout technique-the heat-transfer method. In addition, the design of a miniaturized version of the sensing platform is presented that aims to bridge the gap between measurements in a laboratory setting and point-of-care measurements. This fully automated and integrated, user-friendly design features a capillary pumping unit that is compatible with point-of-care sampling techniques such as a blood lancet device (sample volume-between 50 µL and 300 µL). Sample pre-treatment is limited to the addition of an anti-coagulant. With this fully integrated setup, it is possible to successfully discriminate serotonin from a competitor neurotransmitter (histamine) in whole blood samples. This is the first demonstration of a point-of-care ready device based on synthetic receptors for the screening of neurotransmitters in complex matrices, illustrating the sensor's potential application in clinical research and diagnosis of e.g., early stage depression.

Bystander Effect Induced by Electroporation is Possibly Mediated by Microvesicles and Dependent on Pulse Amplitude, Repetition Frequency and Cell Type.

Bystander effect, a known phenomenon in radiation biology, where irradiated cells release signals which cause damage to nearby, unirradiated cells, has not been explored in electroporated cells yet. Therefore, our aim was to determine whether bystander effect is present in electroporated melanoma cells in vitro, by determining viability of non-electroporated cells exposed to medium from electroporated cells and by the release of microvesicles as potential indicators of the bystander effect. Here, we demonstrated that electroporation of cells induces bystander effect: Cells exposed to electric pulses mediated their damage to the non-electroporated cells, thus decreasing cell viability. We have shown that shedding microvesicles may be one of the ways used by the cells to mediate the death signals to the neighboring cells. The murine melanoma B16F1 cell line was found to be more electrosensitive and thus more prone to bystander effect than the canine melanoma CMeC-1 cell line. In B16F1 cell line, bystander effect was present above the level of electropermeabilization of the cells, with the threshold at 800 V/cm. Furthermore, with increasing electric field intensities and the number of pulses, the bystander effect also increased. In conclusion, electroporation can induce bystander effect which may be mediated by microvesicles, and depends on pulse amplitude, repetition frequency and cell type.

Introducing Thermal Wave Transport Analysis (TWTA): A Thermal Technique for Dopamine Detection by Screen-Printed Electrodes Functionalized with Molecularly Imprinted Polymer (MIP) Particles.

A novel procedure is developed for producing bulk modified Molecularly Imprinted Polymer (MIP) screen-printed electrodes (SPEs), which involves the direct mixing of the polymer particles within the screen-printed ink. This allowed reduction of the sample preparation time from 45 min to 1 min, and resulted in higher reproducibility of the electrodes. The samples are measured with a novel detection method, namely, thermal wave transport analysis (TWTA), relying on the analysis of thermal waves through a functional interface. As a first proof-of-principle, MIPs for dopamine are developed and successfully incorporated within a bulk modified MIP SPE. The detection limits of dopamine within buffer solutions for the MIP SPEs are determined via three independent techniques. With cyclic voltammetry this was determined to be 4.7 × 10(-6) M, whereas by using the heat-transfer method (HTM) 0.35 × 10(-6) M was obtained, and with the novel TWTA concept 0.26 × 10(-6) M is possible. This TWTA technique is measured simultaneously with HTM and has the benefits of reducing measurement time to less than 5 min and increasing effect size by nearly a factor of two. The two thermal methods are able to enhance dopamine detection by one order of magnitude compared to the electrochemical method. In previous research, it was not possible to measure neurotransmitters in complex samples with HTM, but with the improved signal-to-noise of TWTA for the first time, spiked dopamine concentrations were determined in a relevant food sample. In summary, novel concepts are presented for both the sensor functionalization side by employing screen-printing technology, and on the sensing side, the novel TWTA thermal technique is reported. The developed bio-sensing platform is cost-effective and suitable for mass-production due to the nature of screen-printing technology, which makes it very interesting for neurotransmitter detection in clinical diagnostic applications.

Research on water content measurement method based on heat transfer method.

During the measurement of multiphase flow in low yield oil wells, the liquid volume will vary with the operating characteristics of the pumping unit. Using the pulsating characteristics of the up and down strokes of a pumping unit, the flow rate is measured when there is a flow rate on the up stroke, and the water content is measured when the fluid is stationary on the down stroke. In this paper, the heat transfer method is used to measure the water content of the oil water mixture during the down stroke process. At this time, the water content can be expressed as the instantaneous water content of the oil well. Firstly, the feasibility of measuring water content using heat transfer method is demonstrated theoretically, and then the temperature change of the heating probe PT300 is simulated. Finally, the actual temperature of PT300 is measured experimentally. Comparing the experimental value with the simulation value, the calculated measurement error is within 1.27 %, which indicates that the heat transfer method is feasible for measuring water content. Using the same single sensor to measure oil water two-phase flow using the pulsation characteristics of the up and down strokes of a pumping unit is a major innovation in this paper. And lays a foundation for the detection of multiphase flow using heat transfer methods. The successful implementation of the text heat transfer method for measuring water content has broken the previous situation of multiple sensor detection, simplified the structure of multiphase flow instruments, and extended the life of the instrument.

"Immunocytochemistry in Cytology: Myth or Reality": Unraveling the Myth - Immunocytochemistry Applications in Thyroid Lesions.

BACKGROUND: Fine-needle aspiration cytology serves as an important preoperative diagnostic tool for thyroid nodules. Despite its excellent diagnostic accuracy, diagnoses based solely on morphological observation can be challenging. Therefore, various ancillary diagnostic techniques have been applied, including immunocytochemistry (ICC). This review discusses the application and evaluation of ICC in thyroid fine needle aspiration. SUMMARY: Currently, three immunostaining preparation methods are available for cytological materials: liquid-based cytology, cell block, and cell transfer. ICC proves valuable in scenarios such as tumour diagnosis, assessment of differentiation and grading of carcinomas, estimation of primary organs in metastatic carcinomas, and detection of gene abnormalities. However, ICC, while useful, is not as accurate as immunohistochemistry and is more difficult to evaluate. KEY MESSAGES: If the pitfalls and limitations are understood and effectively navigated, ICC could play a significant role in decreasing the non-diagnostic rate, thus leading to more accurate and valuable diagnoses and reductions in the re-aspiration rate.