Predicting Drug-Drug Interactions Involving Rifampicin Using a Semi-mechanistic Hepatic Compartmental Model.

AIMS: To develop a semi-mechanistic hepatic compartmental model to predict the effects of rifampicin, a known inducer of CYP3A4 enzyme, on the metabolism of five drugs, in the hope of informing dose adjustments to avoid potential drug-drug interactions. METHODS: A search was conducted for DDI studies on the interactions between rifampicin and CYP substrates that met specific criteria, including the availability of plasma concentration-time profiles, physical and absorption parameters, pharmacokinetic parameters, and the use of healthy subjects at therapeutic doses. The semi-mechanistic model utilized in this study was improved from its predecessors, incorporating additional parameters such as population data (specifically for Chinese and Caucasians), virtual individuals, gender distribution, age range, dosing time points, and coefficients of variation. RESULTS: Optimal parameters were identified for our semi-mechanistic model by validating it with clinical data, resulting in a maximum difference of approximately 2-fold between simulated and observed values. PK data of healthy subjects were used for most CYP3A4 substrates, except for gilteritinib, which showed no significant difference between patients and healthy subjects. Dose adjustment of gilteritinib co-administered with rifampicin required a 3-fold increase of the initial dose, while other substrates were further tuned to achieve the desired drug exposure. CONCLUSIONS: The pharmacokinetic parameters AUCR and CmaxR of drugs metabolized by CYP3A4, when influenced by Rifampicin, were predicted by the semi-mechanistic model to be approximately twice the empirically observed values, which suggests that the semi-mechanistic model was able to reasonably simulate the effect. The doses of four drugs adjusted via simulation to reduce rifampicin interaction.

Role of Gastrointestinal Microbiota from Crucian Carp in Microbial Transformation and Estrogenicity Modification of Novel Plastic Additives.

Ingestion is a major exposure route for hydrophobic organic pollutants in fish, but the microbial transformation and estrogenic modification of the novel plastic additives by the gut microbiota of fish remain obscure. Using an in vitro approach, we provide evidence that structure-related transformation of various plastic additives by the gastric and intestinal (GI) microbiota from crucian carp, with the degradation ratio of bisphenols and triphenyl phosphate faster than those of brominated compounds. The degradation kinetics for these pollutants could be limited by oxygen and cometabolic substrates (i.e., glucose). The fish GI microbiota could utilize the vast majority of carbon sources in a Biolog EcoPlate, suggesting their high metabolic potential and ability to transform various organic compounds. Unique microorganisms associated with transformation of the plastic additives including genera of Citrobacter, Klebsiella, and some unclassified genera in Enterobacteriaceae were identified by combining high-throughput genetic analyses and metagenomic analyses. Through identification of anaerobic transformation products by high-resolution mass spectrometry, alkyl-cleavage was found the common transformation mechanism, and hydrolysis was the major pathway for ester-containing pollutants. After anaerobic incubation, the estrogenic activities of triphenyl phosphate and bisphenols A, F, and AF declined, whereas that of bisphenol AP increased.

Quorum sensing bacteria in microplastics epiphytic biofilms and their biological characteristics which potentially impact marine ecosystem.

Microplastics (MPs) have been shown to be a new type of pollutant in the oceans, with complex biofilms attached to their surfaces. Bacteria with quorum sensing (QS) systems are important participants in biofilms. Such bacteria can secrete and detect signal molecules. When a signal molecule reaches its threshold level, bacteria with QS systems can perform several biological functions, such as biofilm formation and antibiotic metabolite production. However, the ecological effects of QS bacteria in biofilm as MPs distribute globally with ocean currents are not to be elucidate yet. In this study, polypropylene and polyvinyl chloride were selected for on-site enrichment to acquire microplastics with biofilms. Eight culturable QS bacteria in the resulting biofilm were isolated by using biosensor assays, and their biodiversity was analyzed. The profiles of the N-acyl-homoserine lactones (AHLs) produced by these bacteria were analyzed by using thin-layer chromatography (TLC)-bioautography and gas chromatography and mass spectrometry (GC-MS). Biofilm-forming properties and several biological characteristics, such as bacteriostasis, algal inhibition, and dimethylsulfoniopropionate (DMSP) degradation, were explored along with QS quenching. Results showed that QS bacteria were mainly affiliated with class Alphaproteobacteria, particularly Rhodobacteraceae, followed by class Gammaproteobacteria. TLC-bioautography and GC-MS analyses revealed that seven AHLs, namely, C6-HSL, C8-HSL, 3-oxo-C6-HSL, 3-oxo-C8-HSL, 3-oxo-C10-HSL, and two unidentified AHLs were produced. The QS system equipped bacteria with strong biofilm-forming capacity and may contribute to the keystone roles of Rhodobacteraceae. In addition, QS bacteria may exacerbate the adverse environmental effects of MPs, such as inducing the misfeeding of planktons on MPs. This study elucidated the diversity of QS bacteria in MP-associated biofilms and provided a new perspective of the effect of key membrane-forming bacteria on the marine ecological environment.

Stabilizing Cobalt-free Li-rich Layered Oxide Cathodes through Oxygen Lattice Regulation by Two-phase Ru Doping.

The application of Li-rich layered oxides is hindered by their dramatic capacity and voltage decay on cycling. This work comprehensively studies the mechanistic behaviour of cobalt-free Li1.2 Ni0.2 Mn0.6 O2 and demonstrates the positive impact of two-phase Ru doping. A mechanistic transition from the monoclinic to the hexagonal behaviour is found for the structural evolution of Li1.2 Ni0.2 Mn0.6 O2, and the improvement mechanism of Ru doping is understood using the combination of in operando and post-mortem synchrotron analyses. The two-phase Ru doping improves the structural reversibility in the first cycle and restrains structural degradation during cycling by stabilizing oxygen (O2- ) redox and reducing Mn reduction, thus enabling high structural stability, an extraordinarily stable voltage (decay rate <0.45 mV per cycle), and a high capacity-retention rate during long-term cycling. The understanding of the structure-function relationship of Li1.2 Ni0.2 Mn0.6 O2 sheds light on the selective doping strategy and rational materials design for better-performance Li-rich layered oxides.

Introducing 4s-2p Orbital Hybridization to Stabilize Spinel Oxide Cathodes for Lithium-Ion Batteries.

Oxides composed of an oxygen framework and interstitial cations are promising cathode materials for lithium-ion batteries. However, the instability of the oxygen framework under harsh operating conditions results in fast battery capacity decay, due to the weak orbital interactions between cations and oxygen (mainly 3d-2p interaction). Here, a robust and endurable oxygen framework is created by introducing strong 4s-2p orbital hybridization into the structure using LiNi0.5 Mn1.5 O4 oxide as an example. The modified oxide delivers extraordinarily stable battery performance, achieving 71.4 % capacity retention after 2000 cycles at 1 C. This work shows that an orbital-level understanding can be leveraged to engineer high structural stability of the anion oxygen framework of oxides. Moreover, the similarity of the oxygen lattice between oxide electrodes makes this approach extendable to other electrodes, with orbital-focused engineering a new avenue for the fundamental modification of battery materials.

Microplastics in the Coral Reef Systems from Xisha Islands of South China Sea.

The impacts of microplastics on coral reefs are gaining attention due to findings that microplastics affect coral health. This work investigated the distribution and characteristics of microplastics in the seawater, fish, and corals in 3 atolls from the Xisha Islands of South China Sea. In the seawater samples, microplastics were detected in the outer reef slopes, reef flats, and lagoons with abundances ranging from 0.2 to 11.2, 1.0 to 12.2, and 1.0 to 45.2 items L-1, respectively. Microplastic abundance was 0-12.0 items individual-1 (0-4.7 items g-1) in fish and 1.0-44.0 items individual-1(0.02-1.3 items g-1) in coral. The predominant shape and polymer of microplastics in seawater, fish, and coral were fibrous rayon and polyethylene terephthalate (PET). Microplastic sizes primarily ranged from 20-330 µm in both the seawater and fish, while there were relatively more 1-5 mm microplastics in the corals. The shape, size, color, and polymer type distribution patterns of microplastics in seawater more closely resembled those in fish gills than those in fish gastrointestinal tracts or coral samples. This study shows that microplastics are abundant in these coral reef systems and they are captured by fish or "trapped" by corals.

Response of the sediment geochemistry of the Changjiang River (Yangtze River) to the impoundment of the Three Gorges Dam.

Based on the measurement of major and trace elements in suspended sediments in the low reaches of the Changjiang River during throughout a whole hydrologic year, the origins, seasonal variations, and fluxes of multielements and the human impacts on multielements transport processes have been analyzed along with the influence of weathering in the Changjiang River basin. The results show that most element contents were high in both autumn and winter and low in summer, which was largely caused by the dilution of discharge. Weathering detritus in the Changjiang River basin is the main source of most elements in suspended sediments. However, riverine pollution could bring more loadings of Cd, Pb, As and Zn into river water than it did a few decades ago. The annual average fluxes of Cd, Pb and Zn, which are major contamination elements, to the sea were 179 ±â€¯21 tons/year, 7810 ±â€¯675 tons/year, and 12,000 ±â€¯1320 tons/year, respectively, in which approximately 8.7%, 11.9% and 2.7% of their loadings, respectively, were contributed by pollution inputs. Element exports mainly occurred in the summer (44.4%-57.4%) in the lower part of the Changjiang River. A general relationship between sediment retention and element content suggests a positive feedback mechanism for the decreased number of particles, in which element riverine loadings are reduced due to the enhanced trapping effect by the Three Gorges Dam (TGD). Compared to those in 1980, current element shares of the Changjiang River compared to the global budget have declined due to the construction of the TGD.

Lipids as integral components in mussel adhesion.

Lipids are fundamental components of cells in organisms. Recent studies reveal that lipids are also present in cell-free bioadhesives. Examples include barnacle cement, sea star footprints, hairy and smooth pads of insects and gecko setae. Whether reliance on lipids is universal in bioadhesion is not known. In the present study, we demonstrated, for the first time, the involvement of lipids in mussel adhesion. We extracted, identified and localized lipids in the byssal threads. The lipids were confirmed as fatty acids by gas chromatograpy mass spectrometry. δ13C measurements of the fatty acids in the byssus were also conducted. Results show that byssal fatty acids, with concentrations ranging from 1.10-2.51 mg g-1 by thread dry weight depending on the mussel species, are localized both on the surface of and inside the byssal thread and plaque. Over half of the fatty acids were loosely attached to the surface while a small portion were tightly bound to the byssus. Most of the surface fatty acids disappear within a week of thread deposition. δ13C values of byssal fatty acids show isotope fractionation suggesting that thread fatty acids are derived from the foot. It is possible that fatty acids are key players in expelling water and preparing the substrate surface for adhesion. Using lipids in the adhesion process might be a common strategy for organisms in need of temporary or permanent attachment. The process of lipid participation may be as important as adhesive components for developing more efficient man-made glues.

Glutathione homeostasis and Cd tolerance in the Arabidopsis sultr1;1-sultr1;2 double mutant with limiting sulfate supply.

KEY MESSAGE: Cadmium sensitivity in sultr1;1 - sultr1;2 double mutant with limiting sulfate supply is attributed to the decreased glutathione content that affected oxidative defense but not phytochelatins' synthesis. In plants, glutathione (GSH) homeostasis plays pivotal role in cadmium (Cd) detoxification. GSH is synthesized by sulfur (S) assimilation pathway. Many studies have tried to investigate the role of GSH homeostasis on Cd tolerance using mutants; however, most of them have focused on the last few steps of S assimilation. Until now, mutant evidence that explored the relationship between GSH homeostasis on Cd tolerance and S absorption is rare. To further reveal the role of GSH homeostasis on Cd stress, the wild-type and a sultr1;1-sultr1;2 double mutant which had a defect in two distinct high-affinity sulfate transporters were used in this study. Growth parameters, biochemical or zymological indexes and S assimilation-related genes' expression were compared between the mutant and wild-type Arabidopsis plants. It was found that the mutations of SULTR1;1 and SULTR1;2 did not affect Cd accumulation. Compared to the wild-type, the double mutant was more sensitive to Cd under limited sulfate supply and suffered from stronger oxidative damage. More importantly, under the same condition, lower capacity of S assimilation resulted in decreased GSH content in mutant. Faced to the limited GSH accumulation, mutant seedlings consumed a large majority of GSH in pool for the synthesis of phytochelatins rather than participating in the antioxidative defense. Therefore, homeostasis of GSH, imbalance between antioxidative defense and severe oxidative damage led to hypersensitivity of double mutant to Cd under limited sulfate supply.

Development of an impurity-profiling method for source identification of spilled benzene series compounds by gas chromatography with mass spectrometry: Toluene as a case study.

In this study, an impurity profiling method was established for the source identification of spilled benzene series compounds. Toluene was used as a case study to demonstrate the feasibility of this approach. Gas chromatography with mass spectrometry was applied for identification and quantification of the impurities including ethyl benzene, p-xylene, m-xylene, and o-xylene in toluene. Impurities in toluene were detected at very low levels by applying mass spectrometry in selected-ion monitoring mode. Eight authentic toluene samples collected from different manufacturers were analyzed by the developed gas chromatography with mass spectrometry method to construct the characteristic impurity profiling of toluene. Then, combined with scatter distribution, similarity analysis and t-test, a suite of diagnostic ratios based on the impurity distribution was used for the differentiation of toluene from different sources. Results indicated that scatter distribution method can discriminate the original toluene samples from different manufacturers. Similarity calculation and t-test methods can identify effectively the weathered toluene samples. The proposed impurity profiling method was useful for discrimination between toluene samples from different sources. Statistical analysis of these impurity profiles demonstrated the potential to investigate whether two questioned spilled toluene samples encountered in forensic casework are from the same source.

[Study on fluorescence labeling and determination of polypeptide (PC2~PC6) by high performance liquid chromatography].

This study was based on the thiol groups (-SH) of PC2~PC6, which could be reacted with the Monobromobimane (mBBr), in order to get polypeptide derivatives with fluorescent signal. A new method was developed for measuring the Polypeptides by high performance liquid chromatography with fluorescence detector, then the chromatographic conditions of HPLC was optimized; meawhile the reaction proportion of PCs and mBBr was identified by Trap-MS. The results showed that, the reaction proportion of PCs and mBBr was 1:1, the polypeptide derivatives had good stability; the five compounds separation was better, and the peak time focused on the 16.6~22.0 min; the linear correlation coefficient of PC2, PC3, PC4, PC5 and PC6 was >0.9991, and the limits of quantification were 0.3, 0.05, 0.3, 0.5 and 0.8 mg · L(-1) respectively, the recovery rate was 83.0%-102.0%; the method was reproducible, RSD<2%, this method for measuring the peptide compounds was rapid and accurate.

4-Fluoro-2-(phenyl-amino)-benzoic acid.

The title compound, C13H10FNO2, was obtained by the reaction of 2-bromo-4-fluoro-benzoic acid with aniline. There are two independent mol-ecules, A and B, in the asymmetric unit, with slight conformational differences: the dihedral angles between the aromatic rings are 55.63 (5) and 52.65 (5)°. Both mol-ecules feature an intra-molecular N-H⋯O hydrogen bond. In the crystal, the mol-ecules are linked by pairwise O-H⋯O hydrogen bonds to form A-B acid-acid dimers and weak C-H⋯F inter-actions further connect the dimers.

Nonlinear encoding in diffractive information processing using linear optical materials.

Nonlinear encoding of optical information can be achieved using various forms of data representation. Here, we analyze the performances of different nonlinear information encoding strategies that can be employed in diffractive optical processors based on linear materials and shed light on their utility and performance gaps compared to the state-of-the-art digital deep neural networks. For a comprehensive evaluation, we used different datasets to compare the statistical inference performance of simpler-to-implement nonlinear encoding strategies that involve, e.g., phase encoding, against data repetition-based nonlinear encoding strategies. We show that data repetition within a diffractive volume (e.g., through an optical cavity or cascaded introduction of the input data) causes the loss of the universal linear transformation capability of a diffractive optical processor. Therefore, data repetition-based diffractive blocks cannot provide optical analogs to fully connected or convolutional layers commonly employed in digital neural networks. However, they can still be effectively trained for specific inference tasks and achieve enhanced accuracy, benefiting from the nonlinear encoding of the input information. Our results also reveal that phase encoding of input information without data repetition provides a simpler nonlinear encoding strategy with comparable statistical inference accuracy to data repetition-based diffractive processors. Our analyses and conclusions would be of broad interest to explore the push-pull relationship between linear material-based diffractive optical systems and nonlinear encoding strategies in visual information processors.

Occurrence of microplastics in the Haima cold seep area of the South China Sea.

The pollution of deep-sea microplastics has received increasing attention. As a special ecosystem in the deep sea, the cold seep area is of great significance for studying the distribution of microplastics in the deep sea. In this work, the distribution and characteristics of microplastics in seawater, sediments, and shellfish in the Haima cold seep area and the correlation between the characteristics of microplastics in different media and the type of media were studied. Microplastics were found in all three media. The abundance of microplastics in different samples from the Haima cold seep area ranged 1.8-3.8 items/L for the seawater, 11.47-96.8 items/kg (d.w.) for the surface sediments, and 0-5 items/individual (0-0.714 items/g) for the shellfish. The amount of microplastics ingested by shellfish varied among different species. The microplastics in these three media were mainly fibrous, dark-colored, small-sized rayon, polyethylene terephthalate (PET), and polyethylene (PE). In the correlation analysis of microplastic characteristics among the three media, it was found that the characteristics of microplastics in different media in the same area were closely related, and each pair of variables showed a significant positive correlation (P ≤ 0.05). The distinctive geographical conditions would accelerate the interchange of microplastics among various media. Principal component analysis showed that habitat contribute to microplastic feature differences in shellfish. Differences in correlation were observed between the characteristics of shellfish microplastics in different regions and the characteristics of microplastics in surrounding seawater and sediments.

All-optical complex field imaging using diffractive processors.

Complex field imaging, which captures both the amplitude and phase information of input optical fields or objects, can offer rich structural insights into samples, such as their absorption and refractive index distributions. However, conventional image sensors are intensity-based and inherently lack the capability to directly measure the phase distribution of a field. This limitation can be overcome using interferometric or holographic methods, often supplemented by iterative phase retrieval algorithms, leading to a considerable increase in hardware complexity and computational demand. Here, we present a complex field imager design that enables snapshot imaging of both the amplitude and quantitative phase information of input fields using an intensity-based sensor array without any digital processing. Our design utilizes successive deep learning-optimized diffractive surfaces that are structured to collectively modulate the input complex field, forming two independent imaging channels that perform amplitude-to-amplitude and phase-to-intensity transformations between the input and output planes within a compact optical design, axially spanning ~100 wavelengths. The intensity distributions of the output fields at these two channels on the sensor plane directly correspond to the amplitude and quantitative phase profiles of the input complex field, eliminating the need for any digital image reconstruction algorithms. We experimentally validated the efficacy of our complex field diffractive imager designs through 3D-printed prototypes operating at the terahertz spectrum, with the output amplitude and phase channel images closely aligning with our numerical simulations. We envision that this complex field imager will have various applications in security, biomedical imaging, sensing and material science, among others.

All-optical phase conjugation using diffractive wavefront processing.

Optical phase conjugation (OPC) is a nonlinear technique used for counteracting wavefront distortions, with applications ranging from imaging to beam focusing. Here, we present a diffractive wavefront processor to approximate all-optical phase conjugation. Leveraging deep learning, a set of diffractive layers was optimized to all-optically process an arbitrary phase-aberrated input field, producing an output field with a phase distribution that is the conjugate of the input wave. We experimentally validated this wavefront processor by 3D-fabricating diffractive layers and performing OPC on phase distortions never seen during training. Employing terahertz radiation, our diffractive processor successfully performed OPC through a shallow volume that axially spans tens of wavelengths. We also created a diffractive phase-conjugate mirror by combining deep learning-optimized diffractive layers with a standard mirror. Given its compact, passive and multi-wavelength nature, this diffractive wavefront processor can be used for various applications, e.g., turbidity suppression and aberration correction across different spectral bands.

Association between sugar-free beverage intake and childhood obesity among Chinese children and adolescents.

BACKGROUND: The relationship between sugar-free beverage (SFB) intake and childhood obesity among Chinese children is unknown. OBJECTIVES: To describe the status of SFB consumption among children and adolescents in China and assess the association between SFB intake and different types of obesity. METHODS: The study was based on the baseline data of an ongoing cohort project named Evaluation and Monitoring on School-based Nutrition and Growth in Shenzhen (EMSNGS). Food frequency questionnaires were used to collect information on SFB consumption in 3227 students aged 9-17. Physical and clinical examinations were conducted by trained investigators and clinicians. Multivariable binary logistic regression models were performed to assess the association between SFB intake and general obesity, overweight/obesity, abdominal obesity, metabolically unhealthy overweight (MUOW)/metabolically unhealthy obesity (MUO). RESULTS: The median age of the participants was 13.28 years. Among the participants, 55.2% were boys, and 66.1% were adolescents. The median SFB consumption was 16.67 mL/d. After adjusting for potential confounding factors, each 100 mL increase in daily SFB intake was associated with an increased risk of overweight/obesity (OR = 1.14; 95%CI: 1.06-1.23), abdominal obesity (OR = 1.12; 95%CI: 1.03-1.23), and MUOW/MUO (OR = 1.12; 95%CI: 1.02-1.21), respectively. Stratified analyses showed that family income may have an impact on the association between SFB intake and overweight/obesity (P for interaction = 0.021) and abdominal obesity (P for interaction = 0.031). CONCLUSION: SFB intake was positively associated with childhood obesity in Chinese children, particularly among individuals with high-income families.

All-optical image denoising using a diffractive visual processor.

Image denoising, one of the essential inverse problems, targets to remove noise/artifacts from input images. In general, digital image denoising algorithms, executed on computers, present latency due to several iterations implemented in, e.g., graphics processing units (GPUs). While deep learning-enabled methods can operate non-iteratively, they also introduce latency and impose a significant computational burden, leading to increased power consumption. Here, we introduce an analog diffractive image denoiser to all-optically and non-iteratively clean various forms of noise and artifacts from input images - implemented at the speed of light propagation within a thin diffractive visual processor that axially spans <250 × λ, where λ is the wavelength of light. This all-optical image denoiser comprises passive transmissive layers optimized using deep learning to physically scatter the optical modes that represent various noise features, causing them to miss the output image Field-of-View (FoV) while retaining the object features of interest. Our results show that these diffractive denoisers can efficiently remove salt and pepper noise and image rendering-related spatial artifacts from input phase or intensity images while achieving an output power efficiency of ~30-40%. We experimentally demonstrated the effectiveness of this analog denoiser architecture using a 3D-printed diffractive visual processor operating at the terahertz spectrum. Owing to their speed, power-efficiency, and minimal computational overhead, all-optical diffractive denoisers can be transformative for various image display and projection systems, including, e.g., holographic displays.

Selective Extraction of Critical Metals from Spent Li-Ion Battery Cathode: Cation-Anion Coordination and Anti-Solvent Crystallization.

Owing to continuing global use of lithium-ion batteries (LIBs), in particular in electric vehicles (EVs), there is a need for sustainable recycling of spent LIBs. Deep eutectic solvents (DESs) are reported as "green solvents" for low-cost and sustainable recycling. However, the lack of understanding of the coordination mechanisms between DESs and transition metals (Ni, Mn and Co) and Li makes selective separation of transition metals with similar physicochemical properties practically difficult. Here, it is found that the transition metals and Li have a different stable coordination structure with the different anions in DES during leaching. Further, based on the different solubility of these coordination structures in anti-solvent (acetone), a leaching and separation process system is designed, which enables high selective recovery of transition metals and Li from spent cathode LiNi1/3Co1/3Mn1/3O2 (NCM111), with recovery of acetone. Recovery of spent LiCoO2 (LCO) cathode is also evidenced and a significant selective recovery for Co and Li is established, together with recovery and reuse of acetone and DES. It is concluded that the tuning of cation-anion coordination structure and anti-solvent crystallization are practical for selective recovery of critical metal resources in the spent LIBs recycling.

Virtual histological staining of unlabeled autopsy tissue.

Traditional histochemical staining of post-mortem samples often confronts inferior staining quality due to autolysis caused by delayed fixation of cadaver tissue, and such chemical staining procedures covering large tissue areas demand substantial labor, cost and time. Here, we demonstrate virtual staining of autopsy tissue using a trained neural network to rapidly transform autofluorescence images of label-free autopsy tissue sections into brightfield equivalent images, matching hematoxylin and eosin (H&E) stained versions of the same samples. The trained model can effectively accentuate nuclear, cytoplasmic and extracellular features in new autopsy tissue samples that experienced severe autolysis, such as COVID-19 samples never seen before, where the traditional histochemical staining fails to provide consistent staining quality. This virtual autopsy staining technique provides a rapid and resource-efficient solution to generate artifact-free H&E stains despite severe autolysis and cell death, also reducing labor, cost and infrastructure requirements associated with the standard histochemical staining.