Conventional histological and cytological staining with simultaneous immunohistochemistry enabled by invisible chromogens

Conventional histological stains, such as hematoxylin plus eosin (H&E), and immunohistochemistry (IHC) are mainstays of histology that provide complementary diagnostic information. H&E and IHC currently require separate slides, because the stains would otherwise obscure one another. This consumes small specimen, limiting the total amount of testing. Additionally, performing H&E and IHC on different slides does not permit comparison of staining at the single cell level, since the same cells are not present on each slide, and alignment of tissue features can be problematic due to changes in tissue landscape with sectioning. We have solved these problems by performing conventional staining and IHC on the same slide using invisible IHC chromogens, such that the chromogens are not visible when viewing the conventional stain and the conventional stain is excluded from images of the IHC. Covalently deposited chromogens provided a convenient route to invisible chromogen design and are stable to reagents used in conventional staining. A dual-camera brightfield microscope system was developed that permits simultaneous viewing of both visible conventional stains and invisible IHC chromogens. Simultaneous staining was demonstrated on several formalin-fixed paraffin-embedded tissue specimens using single and duplex IHC, with chromogens that absorb ultraviolet and near infrared light, followed by H&E staining. The concept was extended to other conventional stains, including mucicarmine special stain and Papanicoulou stain, and further extended to cytology specimens. In addition to interactive video review, images were recorded using multispectral imaging and image processing to provide flexible production of color composite images and enable quantitative analysis.

Eco-friendly High-Strength Refractory Concrete Containing Calcium Alumina Cement by Reusing Granite Waste as Aggregate

Besides preventing valuable natural resources from going to waste, using stone waste from stone processing plants in concrete helps reduce environmental pollution and, therefore, offers a convenient route to sustainable development. The present study aims to use granite waste (GW) in high-strength refractory concrete. Sixteen high-strength refractory concrete mixes, including two water-to-binder ratios (W/B = 0.17 and 0.2), two silica-fume-to-binder ratios (SF/B = 0.15 and 0.2), two binder contents (B = 1200 and 1400 kg/m3), and two replacement ratios of silica sand by granite waste (GW/Agg = 0 and 50%) were designed and prepared with high-alumina cement (HAC). The concrete specimens were exposed to 1200 °C. Compressive and flexural strength and scanning electron microscopy (SEM) tests were performed on specimens of concrete mixes before and after heating. It was found that in specimens with high binder content (1400 kg/m3), replacing 50% silica sand with GW (GW/Agg = 50%) in refractory concrete improves compressive and flexural strengths by 3–15 and 4–24% before heating, respectively. It was also shown that using GW to replace silica aggregates in concrete specimens with a 1200 kg/m3 binder content not only did not undermine, but also improved the compressive and flexural strengths of refractory concrete after heating by 20–78% and 15–60%, respectively, as a result of sintering. Meanwhile, in the case of the concrete with 1400 kg/m3 binder content, adding GW exacerbated its loss of compressive and flexural strengths after heating due to little or lack of sintering.

Pyrolysis: A Convenient Route for Production of Eco-Friendly Fuels and Precursors for Chemical and Allied Industries

Thermochemical decomposition of post harvest agro-wastes (biomass) to solid carbonaceous material called as bio-char, condensable vapors (bio-oils and bio-tars) and non-condensable vapors (bio-gas or syn-gas) is referred as pyrolysis. The yield of these products from biomass pyrolysis depends on temperature and other conditions (such as vapor retention time and heating rate) of thermal decomposition in air or oxygen excluded reactor. Bio-char is often used as adsorbent in treatment of water contaminated with dye effluent from textile industry and/or emerging contaminants from other industries. It is also used in production of supercapacitor for energy storage, fertilizer composite and soil amendment for slow release of nutrients for plants and stabilizing pH, enhances water holding and ion exchange capacity of soil. Bio-oils are used for transportation fuels, soaps and other cosmetics production. Bio-tars are also used for transportation fuels but with high heating values and also as organic solvents in chemical, biological and biochemical laboratories. Non-condensable vapors are mostly used as bio-fuels. Products of biomass pyrolysis are potential alternative eco-friendly precursors for chemical and allied industries.

Ex Vivo Evaluation of Egg Yolk IgY Degradation in Chicken Gastrointestinal Tract

Egg yolk antibody (immunoglobulin Y, IgY), due to its unique features (e.g., cost-effectiveness for mass production), is emerging as a promising passive immune agent and alternative to antibiotics to combat infectious diseases, particularly in livestock. Oral administration of egg yolk IgY is the most common and convenient route that has been extensively investigated for controlling enteric pathogens. However, the in vivo stability of egg yolk IgY in the gastrointestinal (GI) tract, a critical issue for the success of this approach, still has not been clearly elucidated. Our recent study showed instability of orally administered egg yolk IgY in chicken GI tract, as demonstrated by both in vivo and ex vivo evidence. To better understand the magnitude and dynamics of instability of egg yolk IgY in vivo, in this study, we conducted comprehensive ex vivo analyses by spiking hyperimmune egg yolk IgY in fresh GI contents collected from five broilers at each sampling age (2, 4, or 6 weeks). The pH in gizzard slightly increased with age from 2.4 to 3.0, while the pH in the small intestine was around 5.8. ELISA analysis indicated that a short time of treatment (30 or 60 min) of IgY with the gizzard contents from the chickens at 2, 4, and 6 weeks of age greatly reduced specific IgY titer by over 8, 6, and 5 log2 units, respectively, when compared with saline control. However, small intestine content only had a mild effect on egg yolk IgY, leading to 1 log2 unit of reduction in IgY titer upon 30 min of treatment. Consistent with these findings, SDS-PAGE and immunoblotting analyses provided direct evidence demonstrating that egg yolk IgY could be drastically degraded to undetectable level in gizzard content upon as short as 5 min of treatment; however, the IgY was only slightly degraded in small intestine content. Immunoblotting also showed that treatment of IgY with HCl (pH 3.0) for 60 min did not affect its integrity at all, further supporting the enzymatic degradation of IgY in gizzard. Collectively, egg yolk IgY could be substantially degraded in chicken gizzard, highly warranting the development of effective approaches, such as encapsulation, for the controlled release and protection of orally administered egg yolk IgY in livestock.

Iterative Upgrading of Small Molecular Tyrosine Kinase Inhibitors for EGFR Mutation in NSCLC: Necessity and Perspective

Molecular targeted therapy has been reported to have fewer adverse effects, and offer a more convenient route of administration, compared with conventional chemotherapy. With the development of sequencing technology, and research on the molecular biology of lung cancer, especially whole-genome information on non-small cell lung cancer (NSCLC), various therapeutic targets have been unveiled. Among the NSCLC-driving gene mutations, epidermal growth factor receptor (EGFR) mutations are the most common, and approximately 10% of Caucasian, and more than 50% of Asian, NSCLC patients have been found to have sensitive EGFR mutations. A variety of targeted therapeutic agents for EGFR mutations have been approved for clinical applications, or are undergoing clinical trials around the world. This review focuses on: the indications of approved small molecular kinase inhibitors for EGFR mutation-positive NSCLC; the mechanisms of drug resistance and the corresponding therapeutic strategies; the principles of reasonable and precision molecular structure; and the drug development discoveries of next-generation inhibitors for EGFR.

A Catalyst-Free Synthesis of Fused Perfluoroalkylated 2,3-Dihydroisoxazoles via Oxa-Michael-Aldol Annulation

A novel synthesis of fused perfluoroalkylated 2,3-dihydroisoxazoles is achieved via oxa-Michael-aldol annulation between perfluoroalk-2-ynoates and N-hydroxyimides. This method provides a convenient route for the synthesis of pyrrolidin-2-one-fused perfluoroalkylated 2,3-dihydroisoxazoles in yields of up to 97%. Diverse and pharmaceutically attractive polycyclic scaffolds can be obtained rapidly and efficiently under these mild, catalyst-free conditions.

Facile Preparation of All Cellulose Self-reinforced Nanocomposites from Bamboo Shoot Fibers

Herein, a novel and facile fabrication method of self-reinforced all cellulose nanocomposite based on 2,2,6,6-tetramethylpiperidine-1-oxy (TEMPO) mediated oxidized bamboo shoot shell fibers was introduced. The composites were thoughtfully characterized. Cellulose nanowhiskers from the bamboo shoot fibers with the diameter of 60–90 nm and a large number of micropores were evenly distributed on the surface of the nanocomposite. Compared with the original fiber, the crystallinity of the composites increased, while the thermal stability decreased. The composite also showed good mechanical property and dimensional stability. It provides a promising and convenient route to obtain firm sheet-materials with micro- or nano- structures from nature cellulose fibers.

The Kuchum Settlement: The Capital of Siberia or a Historical Myth (Archaeological Facts and Historical Assumptions)

Research objectives: The article scrutinizes the arguments challenging the urban status of the Kuchum settlement (Isker), the capital of the Siberian Khanate. The author analyzes the available data on the vicinity, population, trading, and craft production of this capital. Research materials: The main sources are materials obtained during archaeological research in the ancient settlement in the twentieth and twenty-first century, as well as artifacts collected on the shallows of the Irtysh River from the destroyed cultural layer of the mo­nument since the 1880s. The cartographic materials of S.U. Remezov, V. Filimonov, and V.N. Pignatti were also used in the study. Results and novelty of the research: An analysis of the available data showed that the area of the monument reached at least 2.6 hectares by the late fifteenth century. In addition, archaeological research by A.P. Zykov and the author revealed a complex multi-level fortification system aimed at defending the capital of the Siberian Khanate. The artifacts found in 2019 suggest the presence of oversize brick structures in the capital. Such artifacts prove the important role of Isker as the center of trade located on the most convenient route which connected Central Asia, China, Eastern Europe, and the taiga regions of Western Siberia. The population of the settlement widely used silver and copper coins in trading. The collected data show that blacksmiths, casters, jewelers, tailors, and tanners lived and worked in Isker. A huge cultural layer, rich in artifacts, of the Kuchum settlement could be formed only by a fairly large population by Siberian standards, consisting of the khan’s family and its servants, close members of the nobility, Muslim clergy, warriors, merchants, and artisans. The total available data allows us to consider Isker as the political, commercial, craft, and religious center of the Siberian Tatars.

Selective growth of Ti3+/TiO2/CNT and Ti3+/TiO2/C nanocomposite for enhanced visible-light utilization to degrade organic pollutants by lowering TiO2-bandgap

A convenient route was developed for the selective preparation of two stable nanocomposites, Ti3+/TiO2/CNT (labeled as TTOC-1 and TTOC-3) and Ti3+/TiO2/carbon layer (labeled as TTOC-2), from the same precursor by varying the amount of single-walled carbon nanotubes used in the synthesis. TiO2 is an effective photocatalyst; however, its wide bandgap limits its usefulness to the UV region. As a solution to this problem, our prepared nanocomposites exhibit a small bandgap and wide visible-light (VL) absorption because of the introduction of carbonaceous species and Ti3+ vacancies. The photocatalytic efficiency of the nanocomposites was examined via the degradation of methylene blue dye under VL. Excellent photocatalytic activity of 83%, 98%, and 93% was observed for TTOC-1, TTOC-2, and TTOC-3 nanocomposites within 25 min. In addition, the photocatalytic degradation efficiency of TTOC-2 toward methyl orange, phenol, rhodamine B, and congo red was 28%, 69%, 71%, and 91%, respectively, under similar experimental conditions after 25 min. Higher reusability and structural integrity of the as-synthesized photocatalyst were confirmed within five consecutive runs by photocatalytic test and X-ray diffraction analysis, respectively. The resulting nanocomposites provide new insights into the development of VL-active and stable photocatalysts with high efficiencies.