Staining Agent Research Articles

Utility of Lugol’s iodine in assessing the margin status of early stage oral cavity cancer-could it be a simple and effective way to achieve R0 resection?

Background: It is often challenging to distinguish between dysplastic and normal mucosa during surgery. Use of staining agents that will identify abnormal mucosa may improve results. Because Lugol's iodine is readily available, simple to use, and cost-effective, it was chosen as the preferred staining agent. To determine the effectiveness of Lugol’s iodine application in achieving R0 resection in early-stage oral cavity malignancy and thereby leading to better outcomes after treatment. Methods: In this study we included 49 patients and randomized them into two arms. One arm we did wide excision for early-stage oral cavity cancers guided by subjective assessment of margins by surgeons and in another arm, we used Lugol’s iodine to define the margin. We analysed the margin status and thereby the need for the adjuvant therapy between these two groups. Results: In those 49 patients, two of them had microscopically positive margin. One belonged to Lugol’s iodine arm and other belonged to control arm. 17 patients had close margin at mucosal site (less than 5 mm) out of which 11 belonged to control arm and 5 belonged to Lugol’s iodine arm. Conclusions: Lugol’s iodine may be a simple and reliable method in achieving better resection margins and R0 resections for early-stage oral cavity cancer. By using this simple and cost-effective method we may reduce the possibility of ending up with positive or close margins and thereby avoiding the need of chemotherapy and radiation as adjuvant therapy in those patients.

3D Visualization of cyanobacterial biofilms using micro-computed tomography with contrast-enhancing staining agents

Currently, biofilms colonizing surfaces are mainly imaged in 2D by conventional techniques, such as optical or scanning electron microscopy. Confocal laser scanning microscopy or optical coherence tomography can visualize biofilms in 3D, but they suffer from a limited penetration depth and cannot visualize biofilms in opaque materials. Micro-computed tomography (µCT) can overcome these issues, but µCT cannot easily distinguish biofilm structures from water due to a lack of contrast difference. Within this research, five contrast-enhancing staining agents (CESAs) were evaluated for their staining potential of cyanobacterial biofilms, aiming to visualize these biofilms in 3D. Isotonic Lugol and 1:2 hafnium(IV)-substituted Wells-Dawson polyoxometalate (Hf-WD 1:2 POM) were the most promising, as they allowed visualization of the biofilms and revealed structures in the stained biofilms. Staining with isotonic Lugol could clearly visualize bundles of filaments within the biofilm, while Hf-WD 1:2 POM revealed a smooth biofilm. It is assumed that both CESAs have a different affinity towards the biofilms and could thus be used complementary. Monolacunary Wells-Dawson polyoxometalate (Mono-WD POM) showed moderate discrimination while staining with cationic iodinated CA4+ and Hexabrix® (Guerbet) containing anionic ioxaglate did not allow to distinctly visualize the biofilms. These results indicate that µCT, together with CESAs such as isotonic Lugol and Hf-WD 1:2 POM, can be used as a tool to image extensive biofilms or microbial mats in 3D. Further research will determine whether these CESAs are suitable for visualizing biofilms within opaque porous media.

Comparison of two contrast-enhancing staining agents for use in X-ray imaging and digital volume correlation measurements across the cartilage-bone interface

ObjectiveThe pathogenesis of osteoarthritis (OA) is associated with subchondral bone changes, which is linked to abnormal strain distribution in the overlying articular cartilage. This highlights the importance of understanding mechanical interaction at the cartilage-bone interface. The aim of this study is to compare solutions of two contrast-enhancing staining agents (CESA) for combining high-resolution Contrast-Enhanced X-ray microfocus Computed Tomography (CECT) with Digital Volume Correlation (DVC) for full-field strain measurements at the cartilage-bone interface. DesignBovine osteochondral plugs were stained with phosphotungstic acid (PTA) in 70% ethanol or 1:2 hafnium-substituted Wells-Dawson polyoxometalate (Hf-WD POM) in PBS. Mechanical properties were assessed using micromechanical probing and nanoindentation. Strain uncertainties (from CECT data) were evaluated following two consecutive unloaded scans. Residual strains were computed following unconfined compression (ex situ) testing. ResultsPTA and Hf-WD POM enabled the visualisation of structural features in cartilage, allowing DVC computation on the CECT data. Residual strains up to ∼10,000 μɛ were detected up to the tidemark. Nanoindentation showed that PTA-staining caused an average ∼6-fold increase in articular cartilage stiffness, a ∼19-fold increase in reduced modulus and ∼7-fold increase in hardness, whereas Hf-WD POM-stained specimens had mechanical properties similar to pre-stain tissue. Micromechanical probing showed a 77% increase in cartilage surface stiffness after PTA-staining, in comparison to a 16% increase in stiffness after staining with Hf-WD POM. ConclusionHf-WD POM is a more suitable CESA solution compared to PTA for CECT imaging combined with DVC as it allowed visualisation of structural features in the cartilage tissue whilst more closely maintaining tissue mechanical properties.

TPGS-modified Chitosan Nanoparticles of EGFR Inhibitor: Physicochemical and In vitro Evaluation against HepG2 Cell Lines.

Gefitinib (GFN) is an Epithelial Growth Factor Receptor (EGFR) inhibitor, and Food and Drug Administration (FDA) has approved medication to treat lung cancer. However, this investigation aimed to produce and characterize Gefitinib (GFN)-loaded chitosan and soy lecithin nanoparticles (NPs) modified with D-α-tocopheryl polyethylene glycol 1000 succinate mono ester (TPGS) and assess their therapeutic potential against HepG2 liver cell lines. Chitosan, a cationic polymer with biocompatible and biodegradable properties, was combined with soy lecithin to develop the NPs loaded with GFN using a self-organizing ionic interaction methodology. The entrapment efficiency and drug loading were found to be 59.04±4.63 to 87.37±3.82% and 33.46±3.76 to 49.50±4.35%, respectively, and results indicated the encapsulation of GEN in NPs. The pH of the formulations was observed between 4.48-4.62. Additionally, all the prepared NPs showed the size and PDI range of 89.2±15.9 nm to 799.2±35.8 nm and 0.179±0.065 to 0.455±0.097, respectively. The FTIR bands in optimized formulation (GFN-NP1) indicated that the drug might be contained within the NP's core. The SEM photograph revealed the spherical shape of NPs. The kinetic release model demonstrated the combination of diffusion and erosion mechanisms. The IC50 value of GFN and GFN-NP1 formulation against the HepG2 cell lines were determined and found to be 63.22±3.36 μg/ml and 45.80±2.53 μg/ml, respectively. DAPI and PI staining agents were used to detect nuclear morphology. It was observed that the optimized GFN-NP1 formulation successfully internalized and inhibited the growth of HepG2 cells. Hence, it can be concluded that the prepared NPs can be a new therapeutic option for treating liver cancer.

A small-molecule probe to decipher stress-induced ER microenvironments and ER-Golgi communication.

Cellular stress is a crucial factor in regulating and maintaining both organismal and microenvironmental homeostasis. It induces a response that also affects the micropolarity of specific cellular compartments, which is essential for early disease diagnosis. In this contribution, we present a quantitative study of micropolarity changes inside the endoplasmic reticulum (ER) during the G1/S and G2/M phases, using a biocompatible small-molecule fluorophore called ER-Oct. This probe is selectively driven to the ER by its hydrophobicity, and it has the fastest diffusion properties among a series of analogous probes. We found that induced ER stress caused cell cycle arrests leading to an increase in ER micropolarity which is well supported by lambda scanning experiments and fluorescence lifetime imaging microscopy (FLIM) as well. ER-Oct is a versatile staining agent that could effectively stain the ER in various living/fixed mammalian cells, isolated ER, Caenorhabditis elegans, and mice tissues. Furthermore, we used this probe to visualize a well-known biological event, ER to Golgi transport, by live-cell fluorescence microscopy. Our exhaustive investigation of micropolarity using ER-staining dye provides a new way to study ER stress, which could provide a deeper understanding of proteostasis in model systems and even in fixed patient samples.

Risk factors of post-operative optic nerve injury after vitrectomy: a systematic review

Background Vitrectomy is a common procedure used to treat various disorders in the back of the eye. Although it is generally considered safe and effective, there is a risk of complications, including optic nerve damage, which can lead to vision problems. The aim of this study is to determine risk factors of optic nerve injury after vitrectomy for various indications. Methods We conducted a systematic review through literature search via Cochrane, PubMed, Scopus, and Embase, as well as a hand search of relevant journals with the keywords: “(postoperative optic nerve injury) AND (vitrectomy) AND (risk factor)”. The inclusion criteria were: (1) patients with postoperative optic nerve injury after vitrectomy, (2) clinical trial, case-control, or cohort study, (3) analyzing risk factors of optic nerve injury after vitrectomy, (4) published in the last 15 years and conducted in English. For risk of bias assessment, we used the risk of bias (RoB) 2.0 for randomized controlled trials and the ROBINS-E tool for observational studies. Results Thirteen eligible studies were included in this study. We found that damage to the optic nerve can happen either during or after a vitrectomy procedure. This is linked to four main risk factors: removing the internal limiting membrane, having a lower average ocular perfusion pressure, using silicone oil as a tamponade agent, and using ICG as an ILM staining agent. Conclusions Removing the internal limiting membrane, having a lower average ocular perfusion pressure, using silicone oil as a tamponade agent, and using ICG as an ILM staining agent were associated with the occurrence of optic nerve injury following vitrectomy. Our study has limitations, including more retrospective studies than prospective ones and difficulties in combining and analyzing information from individual studies. Additionally, merging data from various studies is challenging due to differences in original research. PROSPERO registration CRD42023453533 (22/08/2023).

A low-cost TICT-based staining agent for identification of microplastics: Theoretical studies and simple, cost-effective smartphone-based fluorescence microscope application

A novel staining agent, (5-(4-(diethylamino)benzylidene)− 1,3-dimethylpyrimidine-2,4,6(1 H,3 H,5 H)-trione) (DDB) was developed for the effective detection of environmentally harmful microplastics. DDB has competitive cost advantages, namely its facile synthesis and high yield, over Nile Red (NR), which is commonly used for microplastic staining. The unique photophysical properties of DDB, including emissive twisted intramolecular charge transfer (TICT) and aggregation-induced emission (AIE), were corroborated via spectroscopic investigations and density functional theory (DFT) calculations. Notably, DDB demonstrated superior selectivity for staining microplastics (polyethylene (PE), polyurethane (PU), polypropylene (PP), polyvinyl chloride (PVC), polystyrene (PS), and polyethylene terephthalate (PET)) over non-plastic materials in water. Furthermore, modulation of the solvent environment during the staining process yielded distinct fluorescence in both the green and red channels for specific types of plastic with the interplay between locally excited (LE) and TICT states. Treatment with 5% ethanol results in the selective staining of PE and PET with the emission of red fluorescence, whereas treatment with 30% ethanol facilitates the selective staining of PU, PVC, and PET with the emission of green fluorescence. Additionally, DDB could selectively stain microplastics in spiked soil and river water samples. Furthermore, a smartphone-based fluorescence microscope was developed at a cost below $100, validating the effective detection of microplastics stained with the newly synthesized DDB. The outcomes of this research demonstrate the potential of DDB as an economical and efficient agent for selective microplastic detection.

Tomography-based investigation on the early-time capillary imbibition in stained foam concrete

In this study, the vertical imbibition of water from foam concretes upward were studied with the assistance of staining, which illustrated the way contrast material spread in specimens during CT scans. The 20 × 20 mm2 bottom face of dried rectangular concrete was adopted, and the test was limited to the first 2.5 h. Using cesium (Cs) or iodine (I) as the staining agent, a new-developed X-ray attenuation method (XRAM) was used to obtain the spatial distribution of water. The result showed that Cs and I are equally trustable in tracing water during the early-time imbibition. Besides, two regions, namely “saturated verge” (near the imbibing face) and “unsaturated transport region” (from an inner area), could be recognized from the imbibition path regarding their variant evolving patterns. The saturated verge from the inlet was of constant width, while the scale of the unsaturated transport region kept increasing during imbibition, and the imbibing rate was similar for all specimens in this study. Moreover, even though the imbibition progress in this study was confirmed a diffusion-controlling progress, a broad imbibition front was revealed from the averaged 1D water content profile, which was mainly attributed to the irregular-shaped “saturated verge”.

Enhanced Photocatalytic Performance of g‐C3N4@MnFe2O4 Nanocomposite for Crystal Violet Dye Degradation under Solar Light

AbstractCrystal violet (CV) dye has been used in the textile industry, as printing ink, biological stain, and antimicrobial agent. CV dye persists in water for a long time and poses a serious threat to humans and aquatic life. In this study, g‐C3N4, MnFe2O4 nanoparticles, and g‐C3N4@MnFe2O4 (80 : 20 wt %) nanocomposite were synthesized via thermal decomposition and chemical co‐precipitation routes. The crystallite size was found 36.31 nm and 30 nm for bare MnFe2O4 nanoparticle and g‐C3N4@MnFe2O4 nanocomposite respectively. The optical band gaps of MnFe2O4, g‐C3N4, and g‐C3N4@MnFe2O4 were 1.8 eV, 2.7 eV, and 2.4 eV, respectively. The photoluminescence intensity decreases in the following order: g‐C3N4>g‐C3N4@MnFe2O4 nanocomposite>MnFe2O4. The degradation efficiency of MnFe2O4, g‐C3N4, and g‐C3N4@MnFe2O4 nanocomposite was found to be 25.13 %, 58.11 %, and 98.42 % respectively, and followed first‐order kinetics. The value of the rate constant for g‐C3N4@MnFe2O4 was 0.022 min−1 which was around ten times higher than g‐C3N4 and MnFe2O4. By adding three scavengers, the degradation capacity of nanocomposite was decreased in the order AA (L‐ascorbic acid)>EDTA (ethylenediaminetetraacetic acid)>TBA (t‐butyl alcohol). A slight reduction in degradation efficiency was observed after five consecutive cycles. Thus, this new finding can be applied to the degradation of CV dye‐contaminated industrial wastewater under solar light.

Color Stability of Resin Hybrid Ceramic Materials in Comparison to Zirconia-Reinforced Lithium Silicate Ceramic.

Objectives: This in vitro study aimed to compare the effect of external staining agents (black tea and grape juice) on the color change of three hybrid ceramics including a new material in comparison to zirconia-reinforced lithium silicate. Materials and Methods: Ten samples of two hybrid dental ceramics including Vita Enamic (Vita Zahnfabrik) and Crystal Ultra (Digital Dental) in addition to a resin nanoceramic, Lava Ultimate (3M ESPE); and a zirconia-reinforced lithium silicate glass ceramic, Vita Suprinity (Vita Zahnfabrik) were prepared and immersed in black tea or grape juice for 30 days. A spectrophotometer was used to evaluate color change. All samples were carefully chosen in A3 shades or their equivalent. Two-way ANOVA and Bonferroni tests were used to analyze the color differences (ΔE00) among the materials and the staining solutions at a significant level of 0.05. Results: Significant color changes were observed in Vita Enamic and Lava Ultimate groups after immersion in both solutions (P<0.001). The least color changes (ΔE00) belonged to the Vita Suprinity (1.34) and Crystal Ultra (1.92) groups, respectively. Conclusion: The color changes of Vita Enamic and Lava Ultimate materials were beyond clinical acceptability. However, the mean color changes of Vita Suprinity and Crystal Ultra groups were clinically acceptable.

Naturalized Dyes: A New Opportunity for the Wood Coloring.

Naturalized dyes (NDs) are innovative and eco-friendly synthetic compounds in which a chromophore is covalently linked to a natural sugar (e.g., lactose). The sugar moiety confers water-solubility and biocompatibility to the dye molecule as a whole. NDs have demonstrated potential application in dyeing textiles and leather. The purpose of this work was to demonstrate that selected NDs can be also applied to dye wood. To that aim, two NDs were tested to color beech and poplar wood. The NDs were applied as a simple aqueous solution or mixed with a waterborne, biogenic staining agent (commercially available Gemma U50). Moreover, the effect of the application of a biogenic waterborne top coat (commercially available Resina Plus U49) was also studied. Different methods were tested to investigate the potential application of these NDs to wood. The dyeing behavior was analyzed in terms of penetration into the substrate, covering capacity and color homogeneity through macro- and microscopic observations and colorimetric measurements. The color fastness to water washout and the color stability to light, in particular by exposing the wooden samples to artificial aging (UV radiations in a Solar Box), were also investigated. The NDs, when used as water solutions, were able to afford a homogeneous coating and a pleasant appearance on the wood surface, as well as a good color fastness to washout with water. Dissolving the dyes in the stain or applying the top coat generally resulted in even better color fastness to washout. However, all the application methods tested showed limited resistance to fading in the Solar Box, which therefore remains a drawback for this type of product.

An Optimized Melanin Depigmentation Method for Histopathological and Immunohistochemical Staining of Formalin-Fixed Paraffin-Embedded Tissues.

Introduction. The difficulty in diagnosis of severe melanocytic lesions is a problem to be overcome in pathological practice. Melanin bleaching is an effective approach to ameliorate melanin disturbances in severely pigmented lesions. Although various methods for improving melanin pigmentation in immunohistochemical staining have been reported, these depigmentation methods still need to be optimized and standardized. In this study, the coloring efficiency of 3,3'-diaminobenzidine (DAB) and alkaline phosphatase (AP) after melanin depigmentation was compared under the automatic immunohistochemical staining platform. Methods. The applicability of the optimized depigmentation method was validated in 10 formalin-fixed paraffin-embedded (FFPE) blocks of ocular melanoma tissues. Specimens were demelaninized with 10% hydrogen peroxide at 60°C for immunohistochemical staining (Melan-A and SOX10), and tissue chromogenic staining was performed with DAB and AP detection systems, respectively. Results. The optimized depigmentation method including immunohistochemistry (IHC) could be completed in 3 h, effectively preserving cell morphology and immunoreactivity. Among these, the color-rendering effect and contrast of AP are better than DAB. Conclusion. This optimized method can effectively remove melanin and improve the accuracy of IHC staining interpretation. AP staining has better visibility and readability without the interference of residual melanin. The comparison results showed that after melanin depigmentation, the immunohistochemical staining agent was replaced with red AP, which avoided the misjudgment caused by brown DAB when melanin depigmentation was incomplete. This improved method can be applied to future histopathological and immunohistochemical staining of melanin-deposited tissues.

Bone analysis using an aggregation‐induced emission‐active iridium complex

AbstractFluorescent analysis of bone provides valuable insights into bone structures. However, conventional dyes suffer from low specificity on bone tissue, small stokes shift, short fluorescent lifetime, and aggregation‐caused quenching effect, which result in low efficacy and artifacts. In this work, we design an aggregation‐induced emission (AIE)‐active iridium(III) complex (Ir‐BP2) as a highly selective, convenient, nondestructiveness, and dual‐mode staining agent for bone analysis. Ir‐BP2 containing phosphonate groups selectively binds to hydroxyapatites, the main component of bone matrix, and exhibits turn‐on AIE phosphorescence with prolonged lifetime. Ir‐BP2 exhibits promising biosafety and offers higher accuracy in staining calcium deposits than conventional Alizarin Red S staining assay when it is employed in real‐time monitoring of osteogenesis differentiation process. A ready‐to‐use staining spray of Ir‐BP2 is fabricated. By using fluorescent imaging and lifetime imaging, Ir‐BP2 staining provides valuable insights into bone microstructure analysis, microdamage diagnosis, and bone growth state identification. Further, Ir‐BP2 is successfully applied on a human spine vertebra for diagnosing bone invasiveness of eosinophilic granuloma, validating its clinical practice. This work presents a powerful tool in bone analysis and will lead to new approaches for the diagnosis and treatment of bone‐related diseases.

Non-destructive 3D characterization of the blood vessel wall microstructure in different species and blood vessel types using contrast-enhanced microCT and comparison with synthetic vascular grafts

To improve the current treatment for vascular diseases, such as vascular grafts, intravascular stents, and balloon angioplasty intervention, the evaluation of the native blood vessel microstructure in full 3D could be beneficial. For this purpose, we used contrast-enhanced X-ray microfocus computed tomography (CECT): a combination of X-ray microfocus computed tomography (microCT) and contrast-enhancing staining agents (CESAs) containing high atomic number elements. In this work, we performed a comparative study based on staining time and contrast-enhancement of 2 CESAs: Monolacunary and 1:2 Hafnium-substituted Wells-Dawson polyoxometalate (Mono-WD POM and Hf-WD POM, respectively) for imaging of the porcine aorta. After showing the advantages of Hf-WD POM in terms of contrast enhancement, we expanded our imaging to other species (rat, porcine, and human) and other types of blood vessels (porcine aorta, femoral artery, and vena cava), clearly indicating microstructural differences between different types of blood vessels and different species. We then showed the possibility to extract useful 3D quantitative information from the rat and porcine aortic wall, potentially to be used for computational modeling or for future design optimization of graft materials. Finally, a structural comparison with existing synthetic vascular grafts was made. This information will allow to better understand the in vivo functioning of native blood vessels and to improve the current disease treatments. STATEMENT OF SIGNIFICANCE: Synthetic vascular grafts, used as treatment for some cardiovascular diseases, still often fail clinically, potentially because of a mismatch in mechanical behaviour between the native blood vessel and the graft. To better understand the causes of this mismatch, we studied the full 3D microstructure of blood vessels. For this, we identified Hafnium-substituted Wells-Dawson polyoxometalate as contrast-enhancing staining agent to perform contrast-enhanced X-ray microfocus computed tomography. This technique allowed to show important differences in the microstructure of different types of blood vessels and in different species, as well as with that of synthetic grafts. This information can lead to a better understanding of the functioning of blood vessels and will allow to improve current disease treatments, such as vascular grafts.

Development of a method to investigate strain distribution across the cartilage-bone interface in guinea pig model of spontaneous osteoarthritis using lab-based contrast enhanced X-ray-computed tomography and digital volume correlation

ObjectiveStrain changes at the cartilage-bone interface play a crucial role in osteoarthritis (OA) development. Contrast-Enhanced X-ray Computed Tomography (CECT) and Digital Volume Correlation (DVC) can measure 3D strain changes at the osteochondral interface. Using lab-based CT systems it is often difficult to visualise soft tissues such as articular cartilage without staining to enhance contrast. Contrast-Enhancing Staining Agents (CESAs), such as Phosphotungstic Acid (PTA) in 70% ethanol, can cause tissue shrinkage and alter tissue mechanics. The aims of this study were, firstly, to assess changes to the mechanical properties of osteochondral tissue after staining with a PTA/PBS solution, and secondly, to visualise articular cartilage during loading and with CECT imaging in order to compare strain across the interface in both healthy and OA joints using DVC. DesignNanoindentation was used to assess changes to mechanical properties in articular cartilage and subchondral bone before and after staining. Hindlimbs from Dunkin-Hartley guinea pigs were stained with 1% PTA/PBS at room temperature for 6 days. Two consecutive CECT datasets were acquired for DVC error analysis. In-situ compression with a load corresponding to 2x body weight was applied, the specimen was re-imaged, and DVC was performed between the pre- and post-load tomograms. ResultsNanoindentation before and after PTA/PBS staining showed similar cartilage stiffness (p < 0.05), however, staining significantly decreased the stiffness of subchondral bone (∼9-fold; p = 0.0012). In severe OA specimens, third principal/compressive (εp3) strain was 141.7% higher and shear strain (γ) was 98.2% higher in tibial articular cartilage compared to non-OA (2 – month) specimens. A 23.1% increase in third principal stain strain and a 54.5% significant increase in the shear (γ) strain (p = 0.0027) was transferred into the mineralised regions of calcified cartilage and subchondral bone in severe OA specimens. ConclusionsThese results indicate the suitability of PTA in PBS as a contrast agent for the visualisation of cartilage during CECT imaging and allowed DVC computation of strain across the cartilage-bone interface. However, further research is needed to address the reduction in stiffness of subchondral bone after incubation in PBS.

Terbutryn causes developmental toxicity in zebrafish (Danio rerio) via apoptosis and major organ malformation in the early stages of embryogenesis

Terbutryn (2-(ethylamino)-4-(tert-butylamino)-6-(methylthio)-1,3,5-triazine) is a substituted symmetrical triazine herbicide used in agricultural fields to prevent undesired vegetation growth by inhibiting photosynthesis in target weeds. Although terbutryn has various benefits, long-term exposure, misuse, or abuse of terbutryn may cause non-target toxicity and severe ecosystem pollution. To provide a detailed description of the embryonic developmental toxicity of terbutryn, zebrafish (Danio rerio) were exposed to 2, 4, and 6 mg/L of terbutryn and the morphological changes, pathological abnormalities, and developmental endpoints were assessed relative to that of a solvent control. The results showed that terbutryn induces a loss of survivability, reduction in body and eye size, and edema in the yolk sac. Through fluorescence microscopy, blood vessels, motor neurons, and liver development were investigated using transgenic zebrafish models based on fluorescently tagged genes (fllk1:eGFP, olig2:dsRed, and L-fabp:dsRed). Furthermore, cell death by apoptosis in zebrafish caused by terbutryn exposure was evaluated via acridine orange staining, which is a selective fluorescent staining agent. To support the preceding results, gene expression alterations caused by terbutryn exposure in zebrafish larvae were assessed. The overall results indicate that exposure to terbutryn induces apoptosis and disrupts organ development. These embryonic developmental toxicity results suggest that terbutryn should be applied in the right areas at the appropriate rates, concentrations, and quantities.

Clinical Observation of Non-Staining Circular Capsulorhexis for Hypermature Cataract

Objective: To explore the circular capsulorhexis method and its clinical effect in phacoemulsification of hypermature cataract without staining. Methods: In this prospective randomized case-control study, 22 patients (22 eyes) with hypermature cataract were randomly divided into a staining agent group (10 cases, 10 eyes) and a non-staining agent group (12 cases, 12 eyes). Both groups underwent phacoemulsification with foldable intraocular lens implantation under topical anesthesia. In the staining agent group, 0.5% indocyanine green was used to stain the anterior lens capsule to assist in circular capsulorhexis; in the non-staining agent group, after puncturing the anterior lens capsule and aspirating the liquefied cortical material, the capsular bag was filled with viscoelastic agent to increase the contrast prior to circular capsulorhexis. The operability of the anterior capsule without staining in capsulorhexis was observed, and the effect of capsulorhexis, diameter of capsulorhexis opening, and operative time were compared and analyzed between the two groups. Results: Both groups successfully underwent circular capsulorhexis, with a success rate of 100%. The diameters of capsulorhexis opening in the staining agent group and non-staining agent group were 5.68 ± 0.15 mm and 5.54 ± 0.16 mm, respectively, without any statistical difference between the groups (P &gt; 0.05); the operative time of the staining agent group (40.40 ± 5.42 s) was shorter than that of the non-staining agent group (50.92 ± 3.97 s), and the difference was statistically significant (P &lt; 0.001). In both groups, phacoemulsification was successful, and intraocular lens were implanted into the capsular bags; their visual acuity showed significant improvement during the 1-month follow-up period after surgery, without any surgical complications. Conclusion: In hypermature cataract, injecting viscoelastic agent into the capsular bag for volume replacement, given a lack of capsule staining agent, can increase the contrast for capsulorhexis and reduce the risk of large capsular bag collapse on the zonular ligament. Thus, it is considered safe and effective. Circular capsulorhexis is conducive to a smooth phacoemulsification procedure.

Protective Impact of Vanillic Acid on Lipid Profile and Lipid Metabolic Enzymes in Diabetic Hypertensive Rat Model Generated by a High-Fat Diet.

Diabetes is the most common component of metabolic syndrome, including abdominal obesity, insulin resistance, hypertension, and dyslipoproteinemia. This study aims to determine whether vanillic acid has antihyperlipidemic properties in diabetic hypertensive rats. For this study healthy male albino Wister rats (180-220 gm) were selected. A 20-week highfat diet (HFD) was given to produce diabetic hypertension in Wister rats. Control and diabetic hypertensive rats were treated with vanillic acid. Vanillic acid effects on lipid profiles (cholesterol, triglycerides, phospholipids, free fatty acids, high-density lipoproteins (HDL)) and lipid metabolizing enzymes LPL, LCAT, and HMG CoA reductase studied by a conventional method. To understand the effect of vanillic acid control, experimental rat lipid and metabolic enzymes were studied and treated and controlled animal liver tissues were observed using the different histology staining agents. Vanillic acid caused considerable lipid profile reductions except for HDL and increased plasma HDL levels. After eight weeks of vanillic acid administration also boosts lipid marker enzyme activity (HMG CoA reductase, LPL, and LCAT). In addition, vanillic acid reduces the accumulation of collagen in liver tissues. These research studies suggest that vanillic acid has antihyperlipidemic effects in diabetic hypertensive rats fed an HFD.

ANALYSIS OF VITAL STAINING: A NARRATIVE REVIEW

Vital staining is a method used in biology and medicine to specically stain living cells or tissues in order to examine their makeup, activity, and functions. The stains used in vital staining can be used to emphasise characteristics like nuclei, mitochondria, or particular proteins. These stains are frequently specic to particular structures or substances within cells. There are many uses for vital staining, ranging from fundamental studies of cell biology and disease processes to clinical diagnosis and monitoring of medical conditions. In order to satisfy the demands of various research questions and clinical applications, novel staining agents and methodologies have been developed over time. Vital staining is still an effective method for analysing living cells and tissues, despite its drawbacks and possible hazards. Even though there are numerous vital stains discovered Toluidine Blue is more commonly used in practice .

Urany-Less Low Voltage Transmission Electron Microscopy: A Powerful Tool for Ultrastructural Studying of Cyanobacterial Cells.

Sample preparation protocols for conventional high voltage transmission electron microscopy (TEM) heavily rely on the usage of staining agents containing various heavy metals, most commonly uranyl acetate and lead citrate. However high toxicity, rising legal regulations, and problematic waste disposal of uranyl acetate have increased calls for the reduction or even complete replacement of this staining agent. One of the strategies for uranyless imaging is the employment of low-voltage transmission electron microscopy. To investigate the influence of different imaging and staining strategies on the final image of cyanobacterial cells, samples stained by uranyl acetate with lead citrate, as well as unstained samples, were observed using TEM and accelerating voltages of 200 kV or 25 kV. Moreover, to examine the possibilities of reducing chromatic aberration, which often causes issues when imaging using electrons of lower energies, samples were also imaged using a scanning transmission electron microscopy at 15 kV accelerating voltages. The results of this study demonstrate that low-voltage electron microscopy offers great potential for uranyless electron microscopy.