Standard Gel Research Articles

Impact of a Bio-Cross-Linking Agent Obtained from Spent Coffee Grounds on the Physicochemical and Thermal Properties of Gelatin/Κ-Carrageenan Hydrogels.

Gelatine hydrogels can be prepared using different cross-linking methods, such as enzymatic, physical or chemical. Unfortunately, in the case of chemical cross-linking, the typically utilized synthetic cross-linkers are harmful to human health and the environment. Therefore, in accordance with the principles of green chemistry and sustainable development, we have obtained compounds for the chemical cross-linking of hydrogel polymers from the processing of spent coffee grounds. In this study, gelatin/κ-carrageenan hydrogels are cross-linked using a bio-cross-linking agent from spent coffee grounds. Their physicochemical and thermal properties are compared with those of standard physical gels. The chemical cross-linking was confirmed based on FT-IR spectra, which demonstrated the formation of new covalent bonds between the oxidized polyphenols included in the extract from the spent coffee grounds and the amide groups present in the gelatine structure. Significant differences were also observed in morphology (SEM images) and other physico-chemical characteristics (gel fraction, swelling ability, hardness). The chemically cross-linked hydrogels in comparison to physically ones are characterized by a better developed porous network, a slightly higher gel fraction (64.03 ± 4.52% as compared to 68.15 ± 0.77%), and a lower swelling ratio (3820 ± 45% as compared to 1773 ± 35%), while TGA results show that they have better thermal stability. The research confirmed the possibility of using the developed natural cross-linking agent in the process of obtaining hydrogel materials based on bio-polymers.

Electrophoretic analysis of proteins from raw materials and commercial diets used in animal feed

In aquaculture, balanced feed accounts for 50% to 70% of the operational costs of cultivating aquatic organisms. This feed is made from various raw materials that serve as protein sources, which significantly influence the final cost of the feed. Therefore, an electrophoretic study was conducted to compare the protein content in different raw materials and commercial diets used in animal feed. Electrophoretic profiles with standard concentration gels (SDS-PAGE) were used to separate protein fractions from animal-origin raw materials, including Earthworm Meals of Eisenia andrei (HLF) and Eisenia fetida (HLT), meat-bone meal (MBM), and fish meal (FM), as well as plant-origin meals, such as Leucaena leaf meal (LLM), corn meal (YCM), and wheat bran (WBM). Commercial diets (A, B, and C) were also evaluated. The protein content of the samples was determined. For electrophoretic analysis, a protease inhibitor cocktail was used, along with a molecular weight standard with a range from 250 to 4 kDa. The molecular weights of the diets ranged between 31.1 and 234.1 kDa, 36.3 and 190.4 kDa, and 4.281 and 95.685 kDa. Some of the protein bands in the analyzed samples (HLF, HLT, FM, YCM, and WBM) were very close to the reference protein bands at 148, 98, 64, 50, and 36 kDa, corresponding to the molecular weight standard used. Electrophoretic characterization provided insight into the different protein fractions, offering valuable information about their potential functional and nutritional properties. Electrophoresis is an essential analytical tool for assessing both the quality and safety of raw materials and foods.

Unveiling a Multiplexed Sensing System for Advanced Biosensing Applications with Thread-Based Electrochemical Sensors

The miniaturization and portability of electrochemical sensing devices play a crucial role in a wide range of applications, including medical diagnostics, environmental monitoring, and point-of-care testing. Integrating a pseudo-reference electrode addresses challenges linked to traditional counterparts, ensuring enhanced portability and miniaturization without performance compromise. Our research delves into the significance of this innovation, emphasizing its impact on signal stability, reproducibility, and overall sensor reliability. The tailored pseudo-reference for compact electrochemical devices contributes to a highly efficient, cost-effective, and user-friendly design, ideal for point-of-care diagnostics and field applications. This work showcases the design, implementation, and performance evaluation of a portable electrochemical sensing system with a pseudo-reference-integrated readout circuit, improving reliability under diverse operating conditions. In the proposed research, we present the design and implementation of a dual-channel configurable multiplexed sensing platform utilizing thread-based electrochemical sensors for continuous potentiometric and amperometric measurements.For the pH sensor, the fabrication process initiates with the cleaning (using isopropyl alcohol) and subsequent plasma treatment of polysorb sutures. The cleaned sutures undergo multiple coatings (4 cycles) with conductive carbon ink, followed by baking at 60°C for 30 to 35 minutes to ensure uniform conductivity. Carbon-coated (CC) sutures undergo electrochemical pre-treatment in a solution of 0.50 M H2SO4 and 0.10 M KCl, followed by electrodeposition with polyaniline (PANI) using potentiometric deposition at 0.80 V in the presence of 0.50 M aniline in a solution containing 1.0 M H2SO4. An insulating polymer layer of 5% polyurethane (PU) is applied at the sensor's center, leaving ends exposed for electrical connections and subsequent hydrogel coating for real sample analysis. The O2 sensor fabrication involves cleaning silver-coated threads, coating with dielectric ink for a sensing region, and UV curing. Two identical threads serve as the cathode and anode, with an intentionally longer anode region as a common reference for the pH sensor. This comprehensive approach contributes to developing a versatile and efficient sensing system for multiplexed measurements, showcasing potential applications in diverse environments.The measurement system includes a signal conditioning circuit (denoted as analog front-end or AFE in Fig. 1) comprising a trans-impedance amplifier (TIA) and an op-amp in the unity-gain configuration: the former is used to convert the current into voltage which originates from the oxygen sensor whereas the latter is used to read out the open-circuit voltage (OCV) for the pH sensor. The outputs of the TIA and the voltage buffer are sequentially selected using a 2:1 multiplexer with a control bit (SEL) before digitizing using an analog-to-digital converter (ADC). There is also a digital-to-analog converter (DAC) generating a bias voltage (VBIAS) of 0.55V for the oxygen sensor as the oxygen electrode requires a certain potential to maximize the dynamic range of the current measurement. The microcontroller handles digital signal processing, which can be connected to a computer for data storage and visualization. The operation of the sensor multiplexing system is validated in standard buffer solutions and gel mediums with different pH and O2 concentrations. The proposed setup demonstrated a sensitivity of ~65 mV/pH for the pH sensor (n=3), and 0.6621 nA/mgL-1 for the O2 sensor (n=3) in a gel medium with common ions and electrolytes. The sensor results were validated against commercial oxygen and pH probes for the same solutions. The developed sensing platform and sensor can precisely monitor and track changes in pH and O2 levels within less than 2 minutes for repeated cycles. This architecture has the potential for extending the low-power multiplexed sensing to multiple biomarkers with an increased number of sensors for ingestible devices, wearable sensing, and cellular agriculture applications. KEYWORDS: Multiplexed sensing, pseudo-reference, pH and oxygen monitoring, miniaturized circuit, thread-based electrochemical sensor Figure 1

Formulation characterization of lecithin organogel as topical drug delivery system for psoriasis: In-vitro permeation and preclinical evaluation.

Psoriasis is a chronic inflammatory and proliferative skin disease that causes pathological skin changes and has a substantial impact on the quality of patient life. Apremilast was approved by the US Food and Drug Administration as an oral medication for psoriasis and is beneficial in mild to moderate conditions for chronic usage. However, 5%-7% of withdrawals were reported due to severe side effects. To address the issue, a localized drug delivery strategy via the topical route may be a viable approach. However, poor physicochemical properties make it vulnerable to passing through the skin, requiring a specialized drug delivery system to demonstrate its full potential via a topical route like lecithin organogel. The formulation was optimized by screening the suitable lecithin type and non-polar solvents based on the gel formation ability of lecithin and the solubility of apremilast in the solvent. The pseudo-ternary diagram was used to optimize the water content required to form the gel. The optimized gel was found to be shear thinning characterized for rheological parameters, in-vitro diffusion studies, and in-vitro skin distribution studies. Preclinical studies in Imiquimod-induced mice showed a better reduction in severity index, cytokine levels, and epidermal hyperplasia from the lecithin organogel group compared to the apremilast oral administration and marketed standard topical gel group. Based on these results, lecithin organogel can be considered a promising approach to deliver molecules like apremilast by topical route in psoriatic-like conditions.

Evaluating Bacterial Nanocellulose Interfaces for Recording Surface Biopotentials from Plants.

The study of plant electrophysiology offers promising techniques to track plant health and stress in vivo for both agricultural and environmental monitoring applications. Use of superficial electrodes on the plant body to record surface potentials may provide new phenotyping insights. Bacterial nanocellulose (BNC) is a flexible, optically translucent, and water-vapor-permeable material with low manufacturing costs, making it an ideal substrate for non-invasive and non-destructive plant electrodes. This work presents BNC electrodes with screen-printed carbon (graphite) ink-based conductive traces and pads. It investigates the potential of these electrodes for plant surface electrophysiology measurements in comparison to commercially available standard wet gel and needle electrodes. The electrochemically active surface area and impedance of the BNC electrodes varied based on the annealing temperature and time over the ranges of 50 °C to 90 °C and 5 to 60 min, respectively. The water vapor transfer rate and optical transmittance of the BNC substrate were measured to estimate the level of occlusion caused by these surface electrodes on the plant tissue. The total reduction in chlorophyll content under the electrodes was measured after the electrodes were placed on maize leaves for up to 300 h, showing that the BNC caused only a 16% reduction. Maize leaf transpiration was reduced by only 20% under the BNC electrodes after 72 h compared to a 60% reduction under wet gel electrodes in 48 h. On three different model plants, BNC-carbon ink surface electrodes and standard invasive needle electrodes were shown to have a comparable signal quality, with a correlation coefficient of >0.9, when measuring surface biopotentials induced by acute environmental stressors. These are strong indications of the superior performance of the BNC substrate with screen-printed graphite ink as an electrode material for plant surface biopotential recordings.

A Crossover Trial Evaluating Coconut Oil as an Alternative to Commercial Ultrasound Gel in Obstetrical Ultrasounds.

Our objective was to evaluate the quality of obstetrical ultrasound images obtained with coconut oil compared with commercial ultrasound gel and to assess patient acceptability. This was a randomized two-period crossover study in which 40 pregnant patients had standard biometry images obtained with both coconut oil and commercial ultrasound gel during their growth or anatomy ultrasound. All images were then rated by two blinded maternal-fetal medicine physicians on quality, resolution, and detail using a 0 to 100 scale. Contrasts obtained from linear mixed models were used to estimate the differences in image parameters between the agents. Participant experience was evaluated with an acceptability survey which included five items measured on a five-point Likert scale. Image quality, as rated by physicians, was found to be equivalent between commercial ultrasound gel and coconut oil. Additionally, there was not a statistically significant difference in image resolution or detail between the two coupling agents. The overall patient experience was significantly lower for commercial ultrasound gel when compared with coconut oil (mean difference = - 5.48, 95% confidence interval = [-6.89, -4.06]). Ultrasound images collected with coconut oil as the coupling agent are equivalent in quality to those collected using commercial ultrasound gel. Patients also preferred the use of coconut oil during their ultrasound, making its use a possible way to improve the patient ultrasound experience. Coconut oil has the potential as an alternative coupling agent that could significantly increase access to ultrasound use in resource-limited settings. · Coconut oil produces quality images during obstetrical ultrasounds.. · Patients prefer the use of coconut oil to standard ultrasound gel during obstetrical ultrasounds.. · Coconut oil is a coupling agent that could increase ultrasound use in resource-limited settings..

Abstract 2795: An investigation to identify the phylogeny of bacterial communities in the pancreatic cancer microbiome by 16SrDNA sequencing and bioinformatic analysis

Abstract The microbiome refers to the diverse community of microorganisms, including bacteria, viruses, fungi, and other microbes, that inhabit various parts of the human body, such as the skin, mouth, gut, and other mucosal surfaces. The role of the microbiome in cancer is a complex and evolving area of research. While much is still being learned, there is evidence suggesting that the microbiome can influence cancer development and progression in several ways. Some bacteria within the microbiome can produce substances that may be either carcinogenic or protective against cancer. For instance, the conversion of certain dietary components by gut bacteria can produce metabolites that may either promote or inhibit cancer development. Different cancers may be associated with specific alterations in the microbiome, for example, the gut microbiome has been extensively studied in relation to colorectal cancer, with certain bacterial species potentially contributing to the development of this cancer. However, the role of microbiome in Pancreatic cancer is still not extensively studied, henceforth, we did a preliminary study to identify novel bacterial communities around pancreatic cancer microbiome by 16SrDNA sequencing in human pancreatic cancer xenograft model mice. Isolating total genomic DNA from the pancreatic model mice feces, 16SrDNA PCR was done using specific primers and the amplicon obtained as PCR product was confirmed by standard DNA gel electrophoresis with a 1 kb standard ladder to be 1.5kb in size universal for all bacterial 16SrDNA. The amplicon was then sent for Next Generation Sequencing at Zymo Research Corporation (CA). The resultant DNA sequences were analyzed by bioinformatics studies and their phylogeny identified by comparing with 16SrDNA data base. Differential abundance of the bacteria’s of the genera Lachnoclostridium, Robinsoniella, Alistipes, Akkermansia and Lactobacillus were observed analyzing the data along with other bacterial communities. We are currently continuing the project to identify more bacterial communities of significance in the pancreatic cancer microbiome and investigating the role and significance of these bacterial communities in pancreatic cancer tumor microenvironment. Acknowledgement This study was done by a joint collaboration between scientists and students of Elizabeth City State University (ECSU) campus of the University of North Carolina Systems and Mayo Clinic, Jacksonville. This research is supported by the NIH-NIGMS grant#T34GM100831-10 to Dr. H. Banerjee and a University of North Carolina General Administration Grant to Dr. Colby Hunter at ECSU. Citation Format: Hirendra N. Banerjee, Colby Hunter, Narendra Banerjee, Karrington Perry, Khadimou Dia, Anasua Banerjee, Santanu Bhattacharya, Hari Krishnareddy Rachamala, Enfang Wang, Shamit Dutta. An investigation to identify the phylogeny of bacterial communities in the pancreatic cancer microbiome by 16SrDNA sequencing and bioinformatic analysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2795.

At-home bleaching with carbamide peroxide with concentrations below 10%: bleaching efficacy and permeability in the pulp chamber.

To evaluate the bleaching efficacy and permeability of hydrogen peroxide (HP) in the pulp chamber of human teeth bleached with lower concentrations of carbamide peroxide gel (4%, 5% and 7% CP). Bleaching gels with lower concentrations were formulated and a commercial standard gel, 10% CP, was used as a reference. Fifty-six human premolars were randomly divided into four groups. Applications of the bleaching gel were made for 3h for 21 days. The bleaching efficacy was evaluated by digital spectrophotometry on 1, 7, 14 and 21 days, with analysis in the ∆Eab, ∆E00 and WID color spaces. The concentration of HP in the pulp chamber was measured in the same periods by UV-Vis spectrophotometry (µg/mL). Two-way repeated analysis of variance (ANOVA) examined bleaching efficacy and HP permeability, followed by Tukey's post-hoc test (α = 0.05). All groups showed significant color changes, with no statistical differences after the second and third week of bleaching (p > 0.05). The 'time' factor was statistically different (p < 0.05), increasing the bleaching efficacy throughout the treatment. The 4% CP group had lower HP levels in the pulp chamber (p < 0.05). The results seem promising, revealing that low concentration gels are as effective as 10% CP with the benefit of reducing the amount of HP in the pulp chamber. Low concentration 4% PC and 5% PC maintains bleaching efficacy, reduces the penetration of HP peroxide into the pulp chamber, and may reduce tooth sensitivity.

Electrochemical Biosensors for the Detection of Viruses: Must-Have Products or Just Science for Publication?

Electrochemical Biosensors for the Detection of Viruses: Must-Have Products or Just Science for Publication?

Characterization of Novel RHD Allele Variants and Their Implications for Routine Blood Group Diagnostics.

The Rh system, including the highly immunogenic D antigen, is one of the clinically most important blood group systems in transfusion medicine. Numerous alleles of the RHD gene are associated with variant RhD phenotypes. In case of Rh incompatibility, some of them can induce hemolytic transfusion reactions and hemolytic disease of the fetus and newborn. Thus, accurate blood group diagnostics are critical for safe transfusion therapy. We characterized phenotypes of four individuals revealing weakened D expression during routine pre-transfusion testing. Standard gel card matrix techniques with monoclonal and polyclonal anti-D antibodies were used for serological typing, complemented using D epitope and antigen density analysis. Genotyping employing PCR with sequence-specific primers, genomic and allele-specific Sanger sequencing and in silico protein analysis were performed. Four novel RHD alleles associated with weak D or partial D phenotypes were identified. One of the mutations is predicted to disrupt the terminal stop codon and result in an elongated translation of the mutant D protein that phenotypically exhibits a loss of D epitopes. Furthermore, a hybrid gene formed with the homologue RHCE gene is described. The presented data enhances the understanding of the Rh system and may contribute to continued advances in blood group diagnostics.

Resting-state EEG recorded with gel-based vs. consumer dry electrodes: spectral characteristics and across-device correlations.

Recordings of electroencephalographic (EEG) rhythms and their analyses have been instrumental in basic neuroscience, clinical diagnostics, and the field of brain-computer interfaces (BCIs). While in the past such measurements have been conducted mostly in laboratory settings, recent advancements in dry electrode technology pave way to a broader range of consumer and medical application because of their greater convenience compared to gel-based electrodes. Here we conducted resting-state EEG recordings in two groups of healthy participants using three dry-electrode devices, the PSBD Headband, the PSBD Headphones and the Muse Headband, and one standard gel electrode-based system, the NVX. We examined signal quality for various spatial and spectral ranges which are essential for cognitive monitoring and consumer applications. Distinctive characteristics of signal quality were found, with the PSBD Headband showing sensitivity in low-frequency ranges and replicating the modulations of delta, theta and alpha power corresponding to the eyes-open and eyes-closed conditions, and the NVX system performing well in capturing high-frequency oscillations. The PSBD Headphones were more prone to low-frequency artifacts compared to the PSBD Headband, yet recorded modulations in the alpha power and had a strong alignment with the NVX at the higher EEG frequencies. The Muse Headband had several limitations in signal quality. We suggest that while dry-electrode technology appears to be appropriate for the EEG rhythm-based applications, the potential benefits of these technologies in terms of ease of use and accessibility should be carefully weighed against the capacity of each given system.

Real-time and quantitative protein detection via polyacrylamide gel electrophoresis and online intrinsic fluorescence imaging

The detection of intrinsic protein fluorescence is a powerful tool for studying proteins in their native state. Thanks to its label-free and stain-free feature, intrinsic fluorescence detection has been introduced to polyacrylamide gel electrophoresis (PAGE), a fundamental and ubiquitous protein analysis technique, to avoid the tedious detection process. However, the reported methods of intrinsic fluorescence detection were incompatible with online PAGE detection or standard slab gel. Here, we fulfilled online intrinsic fluorescence imaging (IFI) of the standard slab gel to develop a PAGE-IFI method for real-time and quantitative protein detection. To do so, we comprehensively investigated the arrangement of the deep-UV light source to obtain a large imaging area compatible with the standard slab gel, and then designed a semi-open gel electrophoresis apparatus (GEA) to scaffold the gel for the online UV irradiation and IFI with low background noise. Thus, we achieved real-time monitoring of the protein migration, which enabled us to determine the optimal endpoint of PAGE run to improve the sensitivity of IFI. Moreover, online IFI circumvented the broadening of protein bands to enhance the separation resolution. Because of the low background noise and the optimized endpoint, we showcased the quantitative detection of bovine serum albumin (BSA) with a limit of detection (LOD) of 20 ng. The standard slab gel provided a high sample loading volume that allowed us to attain a wide linear range of 0.03–10 μg. These results indicate that the PAGE-IFI method can be a promising alternative to conventional PAGE and can be widely used in molecular biology labs.

Wiener Tauberian theorem and half-space problems for parabolic and elliptic equations

&lt;abstract&gt;&lt;p&gt;For various kinds of parabolic and elliptic partial differential and differential-difference equations, results on the stabilization of solutions are presented. For the Cauchy problem for parabolic equations, the stabilization is treated as the existence of a limit as the time unboundedly increases. For the half-space Dirichlet problem for parabolic equations, the stabilization is treated as the existence of a limit as the independent variable orthogonal to the boundary half-plane unboundedly increases. In the classical case of the heat equation, the necessary and sufficient condition of the stabilization consists of the existence of the limit of mean values of the initial-value (boundary-value) function over balls as the ball radius tends to infinity. For all linear problems considered in the present paper, this property is preserved (including elliptic equations and differential-difference equations). The Wiener Tauberian theorem is used to establish this property. To investigate the differential-difference case, we use the fact that translation operators are Fourier multipliers (as well as differential operators), which allows one to use a standard Gel'fand-Shilov operational scheme. For all quasilinear problems considered in the present paper, the mean value from the stabilization criterion is changed: It undergoes a monotonic map, which is explicitly constructed for each investigated nonlinear boundary-value problem.&lt;/p&gt;&lt;/abstract&gt;

Streamlined synthetic assembly of α-chiral CAAC ligands and catalytic performance of their copper and ruthenium complexes.

The unique electronic and steric parameters of chiral cyclic alkyl amino carbene (CAAC) ligands render them appealing steering ligands for enantioselective transition-metal catalyzed transformations. Due to the lack of efficient synthetic strategies to access particularly attractive α-chiral CAACs assessment and exploitation of their full synthetic potential remain difficult. Herein, we report a streamlined strategy to assemble a library of diastereo- and enantiomerically pure CAAC ligands featuring the notoriously difficult to access α-quaternary stereogenic centers. A tailored Julia-Kocienski olefination reagent allows the Claisen-rearrangement to be leveraged as an expedient route to form the synthetically pivotal racemic α-chiral methallyl aldehydes. Subsequent condensation with chiral amines and further cyclization provided a library of diastereomeric mixtures of the targeted ligand precursors. The CAAC salts as well as their corresponding metal complexes are conveniently separable by standard silica gel flash chromatography closing a long-standing accessibility gap in chiral CAAC ligands with proximal α-chirality. The rapid availability of both diastereomers enables testing of the relevance and synergistic effects of two chiral centers on the ligand in catalytic applications. A broad range of metal complexes with copper, gold, rhodium and ruthenium were obtained and structurally analyzed. The catalytic performances of the corresponding chiral CAAC copper and ruthenium complexes were assessed in enantioselective conjugate borylations and asymmetric ring closing metathesis, displaying selectivities of up 95 : 5 er.

Herbal Emulgel Containing Azadirachta indica (Neem) and Nigella Sativa L. (Black Cumin) Oils in Wound Management: Preclinical Investigations

Abstract Context: In wound management, traditional use of plants has received attention by the scientific community, as traditional medicines are a valuable source of affordable, comprehensive medical care. Azadirachta indica A. Juss (Neem) and Nigella sativa Linn (Black cumin) are well known versatile medicinal plants with wide spectrum of biological activity. Methods: This study was undertaken to evaluate the wound healing activity of neem oil (NO) and black cumin oil (BCO) in rats. The oils were standardized using standard procedures. Gas chromatography mass spectrometry analysis and high performance thin layer chromatography fingerprinting of oils were carried out to identify major phytoconstituents present in the oils. The oils were suitably incorporated in carbopol based emulgel formulation, evaluated for physicochemical parameters and stability. Results: The wound healing potential assessed by the measurement of wound contraction and histopathology studies in excision and burn wound models, demonstrated comparable efficacy with standard nano crystalline silver gel (0.002% w/w). This may be attributed to the presence of phytoconstituents such as nimbidin and thymoquinone present in selected oils. Polyunsaturated fatty acid profile of BCO may be responsible for stimulating the release of vascular endothelial growth factor, which plays a significant role in accelerating wound healing process. Conclusion: Topical carbopol 934 base emulgel containing natural oils such as BCO and NO have been successfully prepared for wound management. However, further studies to establish the mechanistic aspects of wound healing needs to be undertaken.

Establishing a Mouse Genotyping Core Facility Based on Automation, High-resolution Melting Analysis, and Purpose-developed MATLAB Applications for Sample Tracking, Data Analysis, and Report Generation

An in-house genotyping facility should aim to be more cost-effective than outsourced service and more reliable than genotyping performed by short-term employees or students of individual research groups. Reliable genotyping allows efficient and economical management of mice colonies and promotes accurate and reproducible research results. Here we provide a detailed description of our approach to establishing a genotyping core facility, relying on automated PCR assembly and high-resolution melting (HRM) analysis (first derivative). The workflow we devised was tightly managed by purpose-designed applications developed using MATLAB App Designer that allowed straightforward work planning, ensured sample tracking throughout the process, and provided a platform for reliable data analysis and generation of genotyping reports. We successfully transitioned PCR product analysis of more than 250 different target genes from standard gel electrophoresis to the more advanced HRM analysis. About 23% of the target genes required a redesign of primers to adapt to our protocol. The process was highly universal, and only 2% of the target genes required deviation from the standard PCR method to a more restricted protocol that reduces the amplification of nonspecific products. We currently run more than 1,000 PCR reactions weekly, of samples taken at weaning or experimental endpoint, and assemble a large variety of target genes in every PCR plate. We also showed that genotyping of blastocytes instead of embryos can serve as quality control of cryopreservation. Thus, our genotyping protocol promotes the 3Rs (Replacement, Reduction, and Refinement) principles. Our refined genotyping process facilitates cost-effective colony management, replaces tissue types as well as traditional methods with advanced ones, and provides reliable results in a timely manner. MATLAB codes and related data are available in supplementary materials and online.

Establishing a Mouse Genotyping Core Facility Based on Automation, High-resolution Melting Analysis, and Purpose-developed MATLAB Applications for Sample Tracking, Data Analysis, and Report Generation.

An in-house genotyping facility should aim to be more cost-effective than outsourced service and more reliable than genotyping performed by short-term employees or students of individual research groups. Reliable genotyping allows efficient and economical management of mice colonies and promotes accurate and reproducible research results. Here we provide a detailed description of our approach to establishing a genotyping core facility, relying on automated PCR assembly and high-resolution melting (HRM) analysis (first derivative). The workflow we devised was tightly managed by purpose-designed applications developed using MATLAB App Designer that allowed straightforward work planning, ensured sample tracking throughout the process, and provided a platform for reliable data analysis and generation of genotyping reports. We successfully transitioned PCR product analysis of more than 250 different target genes from standard gel electrophoresis to the more advanced HRM analysis. About 23% of the target genes required a redesign of primers to adapt to our protocol. The process was highly universal, and only 2% of the target genes required deviation from the standard PCR method to a more restricted protocol that reduces the amplification of nonspecific products. We currently run more than 1,000 PCR reactions weekly, of samples taken at weaning or experimental endpoint, and assemble a large variety of target genes in every PCR plate. We also showed that genotyping of blastocytes instead of embryos can serve as quality control of cryopreservation. Thus, our genotyping protocol promotes the 3Rs (Replacement, Reduction, and Refinement) principles. Our refined genotyping process facilitates cost-effective colony management, replaces tissue types as well as traditional methods with advanced ones, and provides reliable results in a timely manner. MATLAB codes and related data are available in supplementary materials and online.

Roll-to-roll fabrication of silver/silver chloride coated yarns for dry electrodes and applications in biosignal monitoring

This work presents a continuous roll-to-roll electrochemical coating system for producing silver/silver chloride (Ag/AgCl)-coated yarns, and their application in e-textile electrodes for biosignal monitoring. Ag/AgCl is one of the most preferred electrode materials as an interface between the conductive backbone of an electrode and skin. E-textile Ag/AgCl-coated multi-filament nylon yarns offer stable, flexible, and breathable alternatives to standard rigid or flexible film-based Ag/AgCl electrodes. The developed system allows for highly controlled process parameters to achieve stable and uniform AgCl film deposition on Ag-coated nylon yarns. The electrical, electrochemical properties, and morphology of the coated yarns were characterized. Dry electrodes were fabricated and could measure electrocardiogram (ECG) signals with comparable performance to standard gel electrodes. Ag/AgCl e-textile electrodes demonstrated high stability, with low average polarization potential (1.22 mV/min) compared with Ag-coated electrodes (3.79 mV/min), low impedance (below 2 MΩ, 0.1–150 Hz), and are excellent candidates for heart rate detection and monitoring.

Development and Characterization of Econazole Topical Gel.

The purpose of this work was to develop a novel topical formulation of econazole nitrate based on gel that can be easily scaled up in one pot for the potential treatment of fungal and yeast infections. Econazole nitrate, a topical antifungal, is used to treat tinea versicolor, tinea pedis, and tinea cruris. Compared to applying cream or ointment, topical gels offer numerous advantages, one of which is that the drug is released more quickly to the intended site of action. A viscous mixture of propylene glycol, Capmul® MCM C8, methyl and propyl paraben, and econazole nitrate were mixed together before being formulated into the optimized Carbopol® gel bases. The gel's color, appearance, and homogeneity were assessed visually. For every formulation, the drug content, pH, viscosity, spreadability, and gel strength were characterized. The cup plate diffusion method was used to evaluate the anti-fungal activity of the prepared formulations. To assess the behavior of the developed system, studies on in vitro release and mechanism were conducted. The manufactured formulations were transparent, pale yellow, and exhibited excellent homogeneity. The pH of each formulation was roughly 6.0, making them suitable for topical use. The concentration of Carbopol® 940 resulted in a significant increase in viscosity and gel strength but a significant decrease in spreadability. It was demonstrated that the prepared formulations inhibited the growth of Candida albicans and Aspergillus fumigatus. In contrast, the standard blank gel showed no signs of antifungal action. By increasing the concentration of Carbopol® 940, the in vitro release profile of econazole nitrate significantly decreased. Following the Korsmeyer-Peppas model fitting, all formulations exhibited n values greater than 0.5 and less than 1, indicating that diffusion and gel swelling control econazole nitrate release.

Validation of a scanning technique with minimal compression for measuring muscle volume with freehand 3D ultrasound

Freehand 3D ultrasound (3D-US) is a promising technique for measuring muscle volume but it requires gel pads or water tanks to limit probe compression on the skin which makes it hard to use in clinical applications. Our objectives were to measure the effect of different compressions on muscle volume in order to assess the clinical applicability of a minimal compression method for lower limb muscles. 4 muscles of the lower limb on 15 healthy volunteers were scanned with a new commercial freehand 3D-US setup accessible to clinical experimentators. Each muscle was scanned with 3 levels of compression: standard compression, minimal compression and gel pad (method validated against MRI). Volume was calculated using software segmentation tools. Acquisitions and segmentations were done by the same examiner. There was a significant impact of standard compression on volume measurements, but no difference between minimal compression and gel pad. Standard compression underestimated volume with a mean bias of 16 mL. For minimal compression, 75 % of measured differences were below the predefined clinically acceptable limits of 10 mL. Mean bias for this method was 1.1 mL. In conclusion, standard compression in freehand 3D-US induces a systematic bias in volume calculations. But, with a trained examiner and the necessary precautions to minimize compression, this bias could be abolished and become acceptable in clinical applications. When a high accuracy is required, gel pads could still be important to consider.