Confocal Microscopy Research Articles

Deficiency of NLRP3 protects cerebral pericytes and attenuates Alzheimer's pathology in tau-transgenic mice.

Activation of NLRP3-containing inflammasome, which is responsible for IL-1β maturation, has been shown to contribute to Alzheimer's disease (AD)-associated pathogenesis in both APP- and tau-transgenic mice. However, effects of NLRP3 on pericytes and subsequent cerebrovascular pathology in AD remain unknown. NLRP3-deficient and wild-type AD animal models were generated by crossing human P301S tau-transgenic mice and Nlrp3 knockout mice. AD-associated neuroinflammation, tauopathy, vasculature and pericyte coverage in the brain were investigated using immunohistological and molecular biological methods. To investigate how NLRP3 regulates pericyte activation and survival, pericytes from the brains of Nlrp3 knockout and wild-type mice were cultured, treated with IL-1β and H2O2 at different concentrations and analyzed by confocal microscopy and flow cytometry after staining with fluorescently labelled phalloidin, annexin-V and PDGFRβ antibody. Deficiency of NLRP3 (1) reduced Iba-1, GFAP and AT8 antibody-immunoreactive phosphorylated tau-positive cells, without significantly altering transcription of inflammatory genes, (2) preserved cerebral vasculature and pericyte coverage and up-regulated Osteopontin gene transcription, and (3) improved cognitive function in tau-transgenic mice. In cell culture, NLRP3 deficiency prevented pericyte apoptosis. Treatment with IL-1β or H2O2 increased the expression of PDGFRβ in NLRP3-deficient pericytes, but decreased it in NLRP3 wild-type pericytes in a dose-dependent manner. Inhibition of NLRP3 can promote pericyte survival, improve cerebrovascular function, and attenuate AD pathology in the brain of tau-transgenic mice. Our study supports NLRP3 as a novel therapeutic target for Alzheimer's patients.

Origin of dedifferentiated adipocyte-derived cells (DFAT) during ceiling culture in an Adiponectin Cre-Recombinase mouse model.

Dedifferentiated adipose tissue-derived (DFAT) cells represent an attractive source of stem cells for tissue engineering and the potential treatment of several clinical conditions. Our objective was to determine whether DFAT cells originate from mature adipocytes and address whether contamination from the stromal vascular fraction (SVF) could be as a source for these cells. A murine adiponectin-creERT; mT/mG model was used with the excision of the cassette induced by tamoxifen injection for the cells expressing adiponectin (adipoq). This model allows distinguishing mature adipocytes (green fluorescence) from other SVF cell types (red fluorescence) based on the fluorescent protein expressed. Mature adipocytes and SVF cells were isolated from adipose tissues by collagenase digestion. Ceiling cultures were imaged by time-lapse microscopy. Confocal microscopy was used to follow cells over 21 days. Time-lapse microscopy experiments showed liposecretion occurring in mature adipocytes displaying green fluorescence. Confocal imaging allowed the identification of a heterogeneous cell population expressing green but also red fluorescence after 21 days of culture. Asymmetrical division of mature adipocytes was not observed. In conclusion, liposecretion of mature adipocytes is a phenomenon that can be observed in vitro and DFAT cells do originate from mature adipocytes. However, the population of DFAT cells is heterogenous.

Artificial intelligence and real decisions: predictive systems and generative AI vs. emotive-cognitive legal deliberations

IntroductionActivation of NLRP3-containing inflammasome, which is responsible for IL-1β maturation, has been shown to contribute to Alzheimer’s disease (AD)-associated pathogenesis in both APP- and tau-transgenic mice. However, effects of NLRP3 on pericytes and subsequent cerebrovascular pathology in AD remain unknown.MethodsNLRP3-deficient and wild-type AD animal models were generated by crossing human P301S tau-transgenic mice and Nlrp3 knockout mice. AD-associated neuroinflammation, tauopathy, vasculature and pericyte coverage in the brain were investigated using immunohistological and molecular biological methods. To investigate how NLRP3 regulates pericyte activation and survival, pericytes from the brains of Nlrp3 knockout and wild-type mice were cultured, treated with IL-1β and H2O2 at different concentrations and analyzed by confocal microscopy and flow cytometry after staining with fluorescently labelled phalloidin, annexin-V and PDGFRβ antibody.ResultsDeficiency of NLRP3 (1) reduced Iba-1, GFAP and AT8 antibody-immunoreactive phosphorylated tau-positive cells, without significantly altering transcription of inflammatory genes, (2) preserved cerebral vasculature and pericyte coverage and up-regulated Osteopontin gene transcription, and (3) improved cognitive function in tau-transgenic mice. In cell culture, NLRP3 deficiency prevented pericyte apoptosis. Treatment with IL-1β or H2O2 increased the expression of PDGFRβ in NLRP3-deficient pericytes, but decreased it in NLRP3 wild-type pericytes in a dose-dependent manner.DiscussionInhibition of NLRP3 can promote pericyte survival, improve cerebrovascular function, and attenuate AD pathology in the brain of tau-transgenic mice. Our study supports NLRP3 as a novel therapeutic target for Alzheimer’s patients.

Quantification of Sarcoplasmic Reticulum Ca2+ Release in Primary Ventricular Cardiomyocytes.

In the heart, ion channels generate electrical currents that signal muscle contraction through changes in intracellular calcium concentration, i.e., [Ca2+]. The cardiac ryanodine receptor type 2 (RyR2) is the predominant ion channel responsible for increasing intracellular [Ca2+] by releasing Ca2+ from the sarcoplasmic reticulum (SR). Timely Ca2+ release is necessary for appropriate cardiac function, and dysfunction can cause or contribute to life-threatening diseases such as arrhythmia. Quantification of SR-Ca2+ release in the form of sparks and waves can provide valuable insight into RyR2 opening, and factors that influence or regulate channel function. Here, we provide a series of protocols that outline processes for (1) obtaining high-quality isolated cardiomyocytes, (2) preparing samples for experimentally investigating factors that influence RyR2 function, and (3) data acquisition and analysis. Notably, our protocols leverage the potency of the recently developed myosin ATPase inhibitor, Mavacamten. This affords the opportunity to characterize the effects of small molecules or reconstituted proteins/enzymes on RyR2-Ca2+ release events across a range of [Ca2+]. © 2024 Wiley Periodicals LLC. Basic Protocol 1: Cardiomyocyte isolation from mouse Basic Protocol 2: Preparation of cardiomyocytes for Ca2+ imaging Basic Protocol 3: Confocal microscopy and quantitative Ca2+ analysis using SparkMaster 2.

Nicotinic signaling stimulates glucagon secretion in mouse and human pancreatic α-cells.

Smoking is widely regarded as a risk factor for type 2 diabetes, as nicotine contributes to insulin resistance by desensitizing the insulin receptors in muscle, liver, or fat. Little is known, however, about the immediate regulation of islet hormonal output by nicotine, an agonist of ionotropic cholinergic receptors. We investigated this by imaging cytosolic Ca2+ dynamics in mouse and human islets using confocal microscopy and measuring glucagon secretion in response to the alkaloid from isolated mouse islets. Nicotine acutely stimulated cytosolic Ca2+ in glucagon-secreting α-cells but not in insulin-secreting β-cells. The 2.8±0.5-fold (p<0.05) increase in the Ca2+, observed in >70% of α-cells, correlated well with a 2.5±0.3-fold stimulation of glucagon secretion. Nicotine-induced elevation of cytosolic Ca2+ relied on the influx from the extracellular compartment rather than the release of the cation from intracellular depots. Metabotropic cholinergic signaling, monitored at the level of intracellular diacylglycerol, was limited to 69% of α-cells vs 94% of β-cells. We conclude that parasympathetic regulation of pancreatic islet hormone release utilizes different signaling pathways in β- (metabotropic) and α-cells (metabotropic and ionotropic), resulting in the finetuning of the ACh-induced glucagon exocytosis. Sustained nicotinic stimulation is, therefore, likely to attenuate insulin sensitivity by increasing glucagon release.

Sealing Ability of Bioceramic Sealer after Ultrasonic Activation of Different Root Canal Irrigating Solutions

Aim To evaluate and compare the sealing ability of bio ceramic root canal sealer after ultrasonic activation of root canal irrigation solutions 3% NaOCl , 40% Citric acid &amp; Neem leaf extract .  Materials and Methods In this study, 42 mandibular premolar teeth with single roots and canals were divided into three groups for irrigation: 3% NaOCl, 40% citric acid, and Neem leaf extract. After access preparation and glide path creation with a #10 K-file (0.02 taper), the canals were instrumented using a crown-down technique to an ISO size of 30, with saline as the irrigant. Ultrasonically activated irrigation was used, followed by saline in all groups. The canals were obturated with Dentsply Gutta- percha and Bio C sealer, 1mm short of the working length, and sealed with temporary restorations. Teeth were incubated at 37°C for 3 days, and horizontal sections at 3mm from the apex were taken. Confocal laser scanning microscopy measured sealer penetration, and statistical analysis was conducted to compare results between groups.  Results The results suggest that 40% citric acid is the most effective irrigant for enhancing sealer penetration into dentinal tubules at the apical third, both in terms of percentage and depth of penetration, compared to 3% NaOCl and neem leaf extract. This indicates that citric acid may improve the sealing ability of root canal treatments more effectively than the other tested irrigants.  Conclusion The results suggest that 40% citric acid is the most effective irrigant for enhancing sealer penetration into dentinal tubules at the apical third, both in terms of percentage and depth of penetration, compared to 3% NaOCl and neem leaf extract. This indicates that citric acid may improve the sealing ability of root canal treatments more effectively than the other tested irrigants.

In-situ observation of solidification process in β-solidifying γ-TiAl-based alloy

The solidification process of Ti-40Al alloy, a representative β-solidifying γ-TiAl-based alloy, is investigated in-situ using high-temperature laser scanning confocal microscopy (HTLSCM). During the heating process, Ti-40Al alloy exhibits a complete β phase microstructure; however, peritectic α phase is observed during solidification. The occurrence of peritectic α phase is discussed and attributed to the segregation of Al under certain solidification conditions. This finding is further confirmed in the directional solidification of another β-solidifying γ-TiAl-based alloy, TNM alloy, and it provides valuable guidance for controlling the microstructure of β-solidifying γ-TiAl-based alloys.

Squamous Cell Carcinoma: From Diagnosis to Effective Treatment-A Comprehensive Guide

Squamous cell carcinoma (SCC) is the second most common malignant skin cancer, and its occurrence is on the rise globally. The main risk factors are cumulative sun exposure, immunosuppression, and inflammation. Early diagnosis and treatment are crucial to prevent metastasis and improve patient outcomes. Advanced diagnostic techniques, such as dermoscopy and reflectance confocal microscopy (RCM), have improved the accuracy of SCC diagnosis. Dermoscopy allows for better identification and targeted biopsy of suspicious areas, while RCM provides noninvasive and precise monitoring of treatment progress. The treatment of SCC has also advanced significantly in recent years. For patients with severe actinic damage and multiple in situ/low-risk SCC, cancer treatment has shown promising results. This approach involves the use of topical agents to target the tumor and the surrounding areas where cancer may develop. Additionally, novel therapies, such as epidermal growth factor receptor inhibitors and immune checkpoint inhibitors have been developed for locally advanced and metastatic SCC. In conclusion, the incidence of SCC has been increasing globally, and early diagnosis and treatment are crucial to prevent metastasis and improve patient outcomes. Advanced diagnostic techn

The zinc transporter Slc30a1 (ZnT1) in macrophages plays a protective role against attenuated Salmonella.

The zinc transporter Slc30a1 plays an essential role in maintaining cellular zinc homeostasis. Despite this, its functional role in macrophages remains largely unknown. Here, we examine the function of Slc30a1 in host defense using mice models infected with an attenuated stain of Salmonella enterica Typhimurium and primary macrophages infected with the attenuated Salmonella. Bulk transcriptome sequencing in primary macrophages identifies Slc30a1 as a candidate in response to Salmonella infection. Whole-mount immunofluorescence and confocal microscopy imaging of primary macrophage and spleen from Salmonella-infected Slc30a1flag-EGFP mice demonstrate Slc30a1 expression is increased in infected macrophages with localization at the plasma membrane and in the cytosol. Lyz2-Cre-driven Slc30a1 conditional knockout mice (Slc30a1fl/fl;Lyz2-Cre) exhibit increased susceptibility to Salmonella infection compared to control littermates. We demonstrate that Slc30a1-deficient macrophages are defective in intracellular killing, which correlated with reduced activation of nuclear factor kappa B and reduction in nitric oxide (NO) production. Notably, the model exhibits intracellular zinc accumulation, demonstrating that Slc30a1 is required for zinc export. We thus conclude that zinc export enables the efficient NO-mediated antibacterial activity of macrophages to control invading Salmonella.

Low-Intensity Focused Ultrasound-Responsive Phase-Transitional Liposomes Loaded with STING Agonist Enhances Immune Activation for Breast Cancer Immunotherapy.

Background: Pharmacologically targeting the STING pathway offers a novel approach to cancer immunotherapy. However, small-molecule STING agonists face challenges such as poor tumor accumulation, rapid clearance, and short-lived effects within the tumor microenvironment, thus limiting their therapeutic potential. To address the challenges of poor specificity and inadequate targeting of STING in breast cancer treatment, herein, we report the design and development of a targeted liposomal delivery system modified with the tumor-targeting peptide iRGD (iRGD-STING-PFP@liposomes). With LIFU irradiation, the liposomal system exploits acoustic cavitation, where gas nuclei form and collapse within the hydrophobic region of the liposome lipid bilayer (transient pore formation), which leads to significantly enhanced drug release. Methods: Transmission electron microscopy (TEM) was used to investigate the physicochemical properties of the targeted liposomes. Encapsulation efficiency and in vitro release were assessed using the dialysis bag method, while the effects of iRGD on liposome targeting were evaluated through laser confocal microscopy. The CCK-8 assay was used to investigate the toxicity and cell growth effects of this system on 4T1 breast cancer cells and HUVEC vascular endothelial cells. A subcutaneous breast cancer tumor model was established to evaluate the tumor-killing effects and therapeutic mechanism of the newly developed liposomes. Results: The liposome carrier exhibited a regular morphology, with a particle size of 232.16 ± 19.82 nm, as indicated by dynamic light scattering (DLS), and demonstrated low toxicity to both HUVEC and 4T1 cells. With an encapsulation efficiency of 41.82 ± 5.67%, the carrier exhibited a slow release pattern in vitro after STING loading. Targeting results indicated that iRGD modification enhanced the system's ability to target 4T1 cells. The iRGD-STING-PFP@liposomes group demonstrated significant tumor growth inhibition in the subcutaneous breast cancer mouse model with effective activation of the immune system, resulting in the highest populations of matured dendritic cells (71.2 ± 5.4%), increased presentation of tumor-related antigens, promoted CD8+ T cell infiltration at the tumor site, and enhanced NK cell activity. Conclusions: The iRGD-STING-PFP@liposomes targeted drug delivery system effectively targets breast cancer cells, providing a new strategy for breast cancer immunotherapy. These findings indicate that iRGD-STING-PFP@liposomes could successfully deliver STING agonists to tumor tissue, trigger the innate immune response, and may serve as a potential platform for targeted immunotherapy.

Corneal Artificial Endothelial Layer (EndoArt): Literature Review and Our Experience.

Background/Objectives: The objective of this study was to examine the morphological corneal changes and outcomes following the implantation of an artificial endothelial layer (EndoArt) in patients with chronic corneal oedema. Methods: A systematic review of the literature was conducted alongside a detailed analysis of two clinical cases with chronic corneal oedema that were treated using EndoArt. Our experience with these two cases is included to provide practical insights and real-world outcomes. Results: Across the 24 cases reported (including the two presented here), an analysis was possible in 23 cases. Notably, 82% of patients had undergone at least one previous corneal transplant, with 39% having undergone three or more transplants. Additionally, 78% of cases had ocular comorbidities, with glaucoma surgery being the most prevalent (83%), which could have impacted visual outcomes. The follow-up period ranged from 3 to 17 months with a median of 3 months. After EndoArt implantation, the average reduction in the central corneal thickness (CCT) was 29%, and the rebubbling rate was 47.8%, with some cases requiring no rebubbling, while others required it up to 100% of the time. Visual acuity significantly improved from a mean best-corrected visual acuity (BCVA) value of 1.61 ± 0.5 logMAR to 1.07 ± 0.59 logMAR (p < 0.001). The CCT decreased from 771 ± 146 µm to 580 ± 134 µm (p < 0.001). These findings are consistent with our experience. Conclusions: EndoArt shows promise as an alternative treatment for chronic corneal oedema in complex cases where conventional corneal transplantation has failed or carries a high risk of failure. The morphological changes observed using anterior segment optical coherence tomography (OCT) and in vivo confocal microscopy (IVCM) were similar to those reported after endothelial keratoplasty, with the notable exception of the absence of the hyper-reflective donor-host interface.

Could Pistachio (Pistacia vera) Be a Suitable Alternative Crop for Olive-Growing Mediterranean Areas Affected by Xylella fastidiosa subsp. pauca ST53?

In the olive-growing areas of Apulia (southern Italy) where Xylella fastidiosa has caused enormous damage, there is a need to identify alternative crops. These could include pistachio (Pistacia vera L.), but it is critical to define the impact of the bacterium on this crop and what are the main phytosanitary threats for this species in the areas where the bacterium is now endemic. Therefore, we started evaluating infections caused by X. fastidiosa, the fungus Neofusicoccum mediterraneum, and other pathogens on four pistachio cultivars ('Kerman', 'Aegina', 'Lost Hills', and 'Napoletana') grown in areas where X. fastidiosa has been present for a long time. X. fastidiosa was detected only in one orchard (incidence: 18% 'Napoletana' and 55% 'Kerman') out of six surveyed orchards, with low bacterium concentration (1.67 to 5.98 × 103 CFU ml-1) and no symptoms. N. mediterraneum was retrieved in three orchards just on the cultivar Kerman but with high incidence (up to 30%) and infection level quantified as molecular severity (6.82 to 7.43); no other pathogens were detected. The N. mediterraneum representative isolates characterized in this study showed similarity with Spanish and Portuguese isolates. A confocal microscope analysis for this host-pathogen association suggested no differences in plant response to fungal aggression between the cultivars Kerman and Aegina, but just lack of latent inoculum in 'Aegina' plants, pointing to a possible nursery origin of the infection. Waiting for additional targeted experiments to clearly define host response of pistachio cultivars to Xylella spp., this study also points at N. mediterraneum as a potential threat to this tree crop new for the area.

Targeted Magnetic Nanoparticles for Beta-Amyloid Detection

Background/Objectivities: The presence of beta-amyloid plaques is a part of the pathogenesis of Alzheimer’s disease, but there is currently no universally accepted method for magnetic resonance (MR) imaging of the disease. However, it is known that magnetic nanoparticles (MNPs) can improve the T2 contrast in MR images of various targets. Methods: We used cubic MNPs, which were produced by thermal decomposition and then it was covalently bonded to a modified fluorescently labeled tetrapeptide, HAEE-Cy5, for visualizing beta-amyloid plaques. The interaction of MNPs-HAEE-Cy5 and beta-amyloid was determinate by confocal microscopy using SH-SY5Y cell line. Results: MNPs exhibit relatively high relaxivity (approximately 200 mM−1s−1), which is crucial for enhancing target visibility in MR imaging. HAEE provides targeted delivery of MNPs by specifically interacting with beta-amyloid, while the fluorescent label Cy5 enables monitoring the efficacy of the interaction through confocal microscopy. Conclusions: The MNPs modified with HAEE-Cy5 demonstrated excellent binding to beta-amyloid plaques in vitro, as shown by experiments on the SH-SY5Y cell line. These results suggest that the proposed method has potential for use in future MR imaging studies of Alzheimer’s disease.

Effect of various storage media on the physicochemical properties of plasma-treated dental zirconia

This study investigated the effect of various storage media on the physicochemical properties of plasma-treated 3-mol% yttria-stabilized tetragonal zirconia: air, vacuum, deionized water (DIW), and plasma-activated water (PAW). Each group was divided into five subgroups based on storage periods: immediately after NTP irradiation (T0), and after 1 week (T1), 2 weeks (T2), 3 weeks (T3), and 4 weeks (T4). The control group (C) received no treatment. The storage groups were monitored weekly using various analytical techniques, including contact angle measurements, scanning electron microscopy (SEM), focused ion beam (FIB)-SEM, confocal laser scanning microscopy (CLSM), x-ray photoelectron spectroscopy (XPS), and x-ray diffraction (XRD). Our results demonstrate that plasma-treated zirconia surfaces stored in DIW retained or even increased their hydrophilicity due to the formation of hydrogen bonds and preservation of nitrogen functionalities. In contrast, surfaces stored in air exhibited significant hydrophobic recovery. FIB-SEM analysis showed no adverse internal structural changes regardless of storage medium. The roughness of the zirconia surface slightly increased after plasma treatment and was generally retained across all storage groups for 4 weeks, except for the air storage group. This study concludes that storage in DIW effectively preserves the enhanced surface properties of plasma-activated zirconia for up to 4 weeks.

Comparative Study of High-Cycle Fatigue and Failure Mechanisms in Ultrahigh-Strength CrNiMoWMnV Low-Alloy Steels

This study provides a thorough analysis of the fatigue resistance of two low-alloy ultrahigh-strength steels (UHSSs): Steel A (fully martensitic) and Steel B (martensitic–bainitic). The investigation focused on the fatigue behaviour, damage mechanisms, and failure modes across different microstructures. Fatigue strength was determined through fully reversed tension–compression stress-controlled fatigue tests. Microstructural evolution, fracture surface characteristics, and crack-initiation mechanisms were investigated using laser scanning confocal microscopy and scanning electron microscopy. Microindentation hardness (HIT) tests were conducted to examine the cyclic hardening and softening of the steels. The experimental results revealed that Steel A exhibited superior fatigue resistance compared to Steel B, with fatigue limits of 550 and 500 MPa, respectively. Fracture surface analysis identified non-metallic inclusions (NMIs) comprising the complex MnO-SiO2 as critical sites for crack initiation during cyclic loading in both steels. The HIT results after fatigue indicated significant cyclic softening for Steel A, with HIT values decreasing from 7.7 ± 0.36 to 5.66 ± 0.26 GPa. In contrast, Steel B exhibited slight cyclic hardening, with HIT values increasing from 5.24 ± 0.23 to 5.41 ± 0.31 GPa. Furthermore, the martensitic steel demonstrated superior yield and tensile strengths of 1145 and 1870 MPa, respectively. Analysis of the fatigue behaviour revealed the superior fatigue resistance of martensitic steel. The complex morphology and shape of the NMIs, examined using the 3D microstructure characterisation technique, demonstrated their role as stress concentrators, leading to localised plastic deformation and crack initiation.

CIGB-300 internalizes and impairs viability of NSCLC cells lacking actionable targets by inhibiting casein kinase-2 signaling

Overall response rates in advanced Non-Small Cell Lung Cancer (NSCLC) remains low. Thus, novel molecular targets, tailored drugs and/or drug combinations are needed. Casein Kinase-2 (CK2) is a constitutively active and frequently over-expressed enzyme which fosters tumor survival, proliferation and metastasis. By using a clinical-grade and Cell Penetrating Peptide-based inhibitor coined as CIGB-300, we explore the anti-neoplastic effects caused by interruption of CK2 signaling in lung cancer cells lacking EGFR, ALK and ROS mutations. CIGB-300 penetrated and impaired viability and proliferation of Lung Adenocarcinoma (LUAD) (A549, NCI-H522) and Lung Squamous Carcinoma (LUSC) (NCI-H226 and SK-MES-1) cells in a dose-response manner. The differential activity could not be explained by overall peptide uptake or its subcellular distribution, as evidenced by flow cytometry and confocal microscopy. Upon internalization, CIGB-300 interacted with CK2 catalytic subunits (ɑ1/ɑ2) and CK2 substrates, thus impairing phosphorylation of enzyme substrates (CDC37s13, NPM1s125) and downstream proteins (RPS6s325/326). CK2 inhibition induced an early Reactive Oxygen Species (ROS) and mitochondrial membrane depolarization, which predates lung cancer cell death. Finally, intravenous injection of CIGB-300 in a cell line-based xenograft corroborated CIGB-300’s anti-tumor effects and suggested concurrent in situ reductions of CSNK2ɑ subunit and downstream RPS6s235/236 phosphorylation. Overall, CIGB-300 therapeutic hypothesis and antineoplastic effects demonstrated herein, further support the evaluation of this clinical-grade CK2 inhibitor in advanced NSCLC with limited therapeutic options.

Stimulation of Batch Mesophilic Anaerobic Digestion by Cellulose- and Polysaccharide-Derived Polymers in Landfill Leachates

The fate of biobased and biodegradable cellulose-derived plastics in landfills represents an important topic from economic and environmental points of view. Anaerobic digestion is a cost-effective waste-to-energy technology. The behaviour of six polymer types—that is, cellulose (C), cellulose acetate (CA), viscose (V), nanocellulose (NC), acetate textile (AT), and heteropolysaccharide pectin (P)—was studied under anaerobic batch mesophilic conditions in a landfill leachate for 147 days. The cumulative biogas production was as follows: C&gt;V=CA&gt;&gt;AT&gt;&gt;NC=P. Metagenomic analysis revealed notable variations in the proportion of bacterial and archaeal domains with the highest archaeal abundance in the presence of CA (80.2%) and C (78.5%). At the end of digestion, cellulolytic, hydrolytic, and dehydrogenase activities were measured in the intact samples, as well as the liquid and solid fractions, under aerobic and anaerobic conditions. Cellulolytic activity in P was detected only in the pellet, while in NC, activity was mostly in the supernatant under both aerobic and anaerobic conditions. Scanning electron microscopy and confocal scanning laser microscopy showed a defragmentation and degradation of polymeric substrates as well as microbial colonisation. Based on the results, landfill leachate is appropriate for the anaerobic biodegradation of cellulose-derived polymers; however, the process is polymer specific.

Two-Dimensional MXene Flakes with Large Second Harmonic Generation and Unique Surface Responses.

MXenes are a family of two-dimensional (2D) materials with broad and varied applications in biology, materials science, photonics, and environmental remediation owing to their layered structure and high surface area-to-volume ratio. MXenes have exhibited significant nonlinear optical characteristics, which have been primarily explored in the context of photonics applications, yet the second-harmonic generation (SHG) behavior of MXenes remains an unexplored aspect of their optical properties. Herein, we demonstrate and quantify large second-order responses of 2D Ti3C2Tx MXenes both in aqueous solutions and on a silicon substrate for the first time. MXene flakes showed strong second-harmonic scattering (SHS) in a dilute suspension with a sensitivity of less than 0.1 μg/mL. Angle-dependent SHS experiments further found that the second-order responses originate from coherent 2D dipole radiation. Through confocal and atomic force microscopies, we found that the intense SHG signal from free-standing MXene flakes increases exponentially with decreasing thickness, while two-photon fluorescence increases linearly with thickness. The second-order susceptibility of the MXenes was determined to be 3.6 pm V-1 with a thickness of 10 nm, almost twice of that for an often-used SHG crystal, beta barium borate. We further explored surface properties of the MXene sheets by investigating the SHS responses upon addition of organic dye molecules to the system. It was found that the adsorption of crystal violet (CV) obeys a Langmuir adsorption model while the addition of malachite green (MG) resulted in almost no change in SHG intensity, even though the adsorption capacities for both CV (61.3 ± 1.7 mg/g) and MG (54.8 ± 2.8 mg/g) are similar. Such a stark difference in adsorption characteristics between cationic organic CV and MG dyes is likely due to their distinct orientational orderings on the MXene surfaces. This work opens many possibilities for the further employment of the family of 2D materials in photonics, optics, and surface catalysis applications.

Simple Epithelial Transplantation for Ocular Surface Reconstruction After Severe Ocular Burn Injury.

To manage ocular surface complications and recover conjunctival and corneal epithelia after unilateral severe chemical burn. We performed simple conjunctival epithelial transplantation (SCET) to obtain renewal of fornix and bulbar-tarsal conjunctiva epithelium, followed by simple limbal epithelial transplantation (SLET) to recover limbal function and epithelial corneal surface. Slit-lamp examination, fluorescein staining, in vivo confocal microscopy, Kheirkhah grading system for symblepharon severity, Wong-Baker FACES Pain Rating Scale, and best-corrected visual acuity were assessed before surgery, at 1 to 3 months after SCET and SLET, and thereafter at 6 to 12 to 36 months. Two patients with unilateral burn injuries underwent surgery. Eye mobility and fornix reconstruction were promptly achieved, and conjunctival epithelium with goblet cells was observed on the bulbar and tarsal conjunctiva 3 months after SCET. After SLET, corneal epithelium and cornea-conjunctiva transition zone were observed at 3 and 6 months, respectively. From before surgery to 6 months after SLET, symblepharon improved from grade IVa2 and IIIb2 to Ic0 and Ib0, the Wong-Baker FACES Pain Rating Scale changed from grade 6 and 4 to 0, and best-corrected visual acuity upgraded from 1.40 and 1.10 logarithm of the minimum angle of resolution to 0.5 logarithm of the minimum angle of resolution, in patient 1 and 2, respectively. After 3 years, results remained stable. SCET effectively healed the bare conjunctival area relieving subjective symptoms and discomfort. Sequential SCET and SLET showed to be feasible in restoring a normal ocular surface with long-lasting results suggesting the aim in patients with severe ocular burn is not merely corneal epithelium renewal but also the regeneration of ocular surface homeostasis.

Experimental investigation of the interplay between transverse mixing and pH reaction in porous media

Abstract. pH-induced reactive transport in porous environments is a critical factor in Earth sciences, influencing a range of natural and anthropogenic processes, such as mineral dissolution and precipitation, adsorption and desorption, microbial reactions, and redox transformations. These processes, pivotal to carbon capture and storage (CCS) applications to groundwater remediation, are determined by pH transport. However, the uncertainty in these macroscopic processes’ stems from pore-scale heterogeneities and the high diffusion value of the ions and protons forming the pH range. While practical for field-scale applications, traditional macroscopic models often fail to accurately predict experimental and field results in reactive systems due to their inability to capture the details of the pore-scale pH range. This study investigates the interplay between transverse mixing and pH-driven reactions in porous media. It focuses on how porous structure and flow rate affect mixing and chemical reaction dynamics. Utilizing confocal microscopy, the research visualizes fluorescently labeled fluids, revealing variations in mixing patterns from diffusive in homogeneous to shear-driven in heterogeneous media. However, pH-driven reactions show a different pattern, with a faster reaction rate, suggesting quicker pH equilibration between co-flowing fluids than predicted by transverse dispersion or diffusion. The study highlights the unique characteristics of pH change in water, which significantly influences reactive transport in porous media.