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Background Materials and Methods Results Conclusion Aging is a physiological phenomenon in the process of life, and skin aging has a significant impact on human appearance. Therefore, the search for methods to delay skin aging is of great significance for improving the quality of human life.This study investigated the anti‐photoaging effect of Tricholoma matsutake (T) extract composition combined with bakuchiol (B) and ergothioneine (E), and explored its potential mechanism through transcriptome, metabolomics, and network pharmacology.57 main chemical components are identified from the ethanol extract of T. matsutake (T), including D‐carnitine (24.55%), α,α‐trehalose (15.56%), DL malic acid (8.99%), D‐(−)‐quinic acid (7.46%), erucamide (7.04%) and so on. After TBE treatment, inflammation of the mice dorsal skin is significantly minimized. Hematoxylin and eosin (H&E) staining and toluidine blue staining reveal that TBE has an anti‐inflammatory effect on the back skin tissue of mice. Masson staining shows that TBE has a repair effect on mice dorsal skin tissue. In addition, the inflammatory factors (IL‐1β, IL‐6, TNF‐α) in the mice dorsal skin tissues are significantly reduced but collagen (COL‐1) is significantly increased. By cellular immunofluorescence assay, TBE is shown to promote PPAR‐α expression in cells. Transcriptomics, metabolomics, and network pharmacology have revealed that TBE can regulate exogenous stimuli and cancer‐related signaling pathways to prevent skin aging.The results suggest that TBE can be a beneficial supplement to natural anti‐aging. [ABSTRACT FROM AUTHOR]

Nitric oxide (NO) and electromagnetic fields (EMF) have been extensively studied for their roles in neurobiology, particularly in regulating cerebral functions and synaptic plasticity. This study investigates the impact of EMFs on NO modulation and its subsequent effects on neurodevelopment, building upon prior research examining EMF exposure's consequences on Wistar albino rats. Rats were exposed perinatally to either tap water, 1 g/L of L‐arginine (LA) or 0.5 g/L of N‐methylarginine (NMA). Half of the rats in each group were also exposed to a 7‐Hz square‐wave EMF at three separate intensities (5, 50 and 500 nT) for 2–14 days following birth. Animals were allowed to develop, and their brains were harvested later in adulthood (mean age = 568.17 days, SD = 162.73). Histological analyses were used to elucidate structural changes in key brain regions. All brains were stained with Toluidine Blue O (TBO), enabling the visualization of neurons. Neuronal counts were then conducted in specific regions of interest (e.g. hippocampus, cortices, amygdala and hypothalamus). Histological analyses revealed significant alterations in neuronal density in specific brain regions, particularly in response to EMF exposure and pharmacological interventions. Notable findings include a main EMF exposure effect where increased neuronal counts were observed in the secondary somatosensory cortex under low EMF intensities (p < 0.001) and sex‐specific responses in the hippocampus, where a significant increase in neuronal counts was observed in the left CA3 region in female rats exposed to EMF compared to unexposed females (t(18) = 2.371, p = 0.029). Additionally, a significant increase in neuronal counts in the right entorhinal cortex was seen in male rats exposed to EMF compared to unexposed males (t(18) = 2.216, p = 0.040). These findings emphasize the complex interaction among sex, EMF exposure and pharmacological agents on neuronal dynamics across brain regions, highlighting the need for further research to identify underlying mechanisms and potential implications for cognitive function and neurological health in clinical and environmental contexts. [ABSTRACT FROM AUTHOR]

This is the first study to describe the subtypes, number and distribution of mast cells (MC) in cat tongue by histochemical and immunohistochemical methods. Six male adult felines' tongue tissue samples consist of the study's material. Samples were fixed in 10% formaldehyde. MC number and distribution in the feline tongue were assessed using toluidine blue. Also, sections taken from blocks were stained in alcian blue/safranin O (AB/SO) combined dyes to determine the MC subtypes. The Streptavidin biotin complex method using anti‐chymase and anti‐tryptase primary antibodies was used for immunohistochemistry. Metachromatic MCs were mainly observed in the lamina propria close to the multilayered keratinized stratified squamous epithelium. The high number of MCs in this region may be because the dorsal surface of the tongue plays an essential role in the defence system of tongue tissue and, thus, of the body as a whole. Additionally, the number of MCs stained with AB (+) (1.7 ± 0.08) in the feline tongue was statistically higher than those with SO (+) (0.18 ± 0.02). This might be interpreted as an indication that MC heterogeneity may be due not only to their staining properties but also to their localization. It is also conceivable that the high histamine content may be a factor in this. Tryptase‐positive MCs were found in the loose connective tissue around blood vessels, between the glands, as solitary cells, or in groups of several cells. Chymase‐positive MCs were observed more individually rather than in groups. Moreover, chymase‐positive MCs were detected to be located in the filiform papillae subepithelial and in the blood vessels' immediate vicinity. Animals often lick themselves to clean themselves and promote healing. For this reason, it is very important to protect the tongue, which is in direct contact with the external environment, against foreign agents. Considering both the functional and protective properties of the tongue, we concluded that MCs may play a role in oral cavity immunity and protective effect. [ABSTRACT FROM AUTHOR]

Weakness was observed in the F1 hybrids of crosses between Capsicum annuum × Capsicum chinense or C. annuum × Capsicum frutescens. This phenomenon is morphologically characterized by the cessation of new leaf development approximately 30 days after germination (DAG). We wondered why this phenomenon appeared in the middle of growth and studied the relationship between Capsicum hybrid weakness and the juvenile‐to‐adult (JA) phase transition. Traits that changed during the early vegetative phase in both parents were the petiole/leaf length ratio, number of leaf lateral veins and stomata, colour of epidermal cells in leaves stained with toluidine blue and expression of miR156 and miR172. Based on the changes observed during the early vegetative phase, C. annuum and C. chinense were transferred to the adult phase 28 DAG. However, the F1 hybrids did not show characteristics of the adult phase at any time point. Furthermore, the phenotypes of hybrid weakness were observed at approximately 35 DAG, when F1 hybrids were presumed to reach the JA transition phase. It has been suggested that the JA transition is involved in Capsicum hybrid weakness. [ABSTRACT FROM AUTHOR]

Background: Chemotherapy‐induced peripheral neuropathy (CIPN) is still an unresolved problem in cisplatin (CIS) use. Objectives: This study investigates possible anti‐neuropathic effect of chlorogenic acid (CGA) against CIS‐induced CIPN in rats while also investigating the contribution of nitric oxide (NO) to this phenomenon. Methods: Initially, CGA (250–1000 μM) was tested by MTT assay on primary DRG neurons. Subsequently, CIPN was induced in Sprague–Dawley rats by 3 mg/kg intraperitoneal injections of CIS once/week for 5 weeks. CGA (100 mg/kg) was co‐administered with CIS, both alone and in combination with l‐arginine (LARG) or l‐nitro‐arginine‐methyl‐ester (LNAME), to elucidate the contribution of nitrergic system to anti‐neuropathic effects. Mechanical allodynia, thermal hyperalgesia, and cold plate tests were performed to test CIPN. Rotarod, footprint analysis, and activitymeter were used to evaluate motor coordination and performance. Tumor necrosis factor alpha (TNF‐α) was measured as a marker of inflammation. Histological evaluations of DRG and sciatic nerves (SNs) were performed utilizing toluidine blue staining. Two‐way analysis of variance and Kruskal–Wallis following Tukey's test were used as statistical analysis. Results: Higher concentration of CGA (1000 μM) exhibited protective effect against in vitro neurotoxicity. Neither LARG nor LNAME exerted significant change in this effect. Co‐administration of CGA alleviated histological abnormalities and neuropathic effects induced by CIS. Ameliorative effect of CGA was not changed in mechanical allodynia but attenuated in cold allodynia, and motor activity/coordination tests by LARG and LNAME. Neuropathic effects of CIS remained unchanged with LARG and LNAME in behavioral experiments. Conclusion: The study identified CGA as candidate agent in mitigating CIPN. NO seems to play a modulatory role in this effect. [ABSTRACT FROM AUTHOR]

Background: Research has demonstrated the anti‐photoaging properties of glabridin and bakuchiol. Methods: The impact of glabridin, glabridin + bakuchiol, and bakuchiol on the levels of tumor necrosis factor alpha (TNF‐α) and interleukin‐1 beta (IL‐1β) in mice skin fibroblasts was observed. Furthermore, we investigated the potential roles of fibronectin (FN), interferon‐γ (IFN‐γ), interleukin‐22 (IL‐22), and transforming growth factor‐β (TGF‐β) in the tissues, and evaluated their impact on the enzymatic levels in the skin. In conjunction with transcriptomic analysis, metabolomic profiling, and network pharmacology, all samples underwent comprehensive metabolomic and principal component analysis. The Venny2.1 method was utilized to identify variances in shared metabolites between the treatment group and the UVB group, as well as between the UVB group and the control group. Subsequently, a cluster heat map was generated to forecast and analyze metabolic pathways and targets. Results: The outcomes from the hematoxylin and eosin and toluidine blue staining revealed that glabridin and bakuchiol markedly decreased dermal thickness and suppressed mast cell infiltration in photoaged mice. Immunohistochemistry and Elisa analysis revealed that glabridin and bakuchiol effectively attenuated the levels of pro‐inflammatory factors, including IL‐1β, tumor necrosis factor‐α, IL‐22, and IFN‐γ. Furthermore, an increase in the levels of anti‐inflammatory factors such as FN and TGF‐β was also observed. The determination of the contents of superoxide dismutase, hydroxypropyltransferase and malondialdehyde in mice dorsal skin revealed that glabridin and bakuchiol not only elevated the levels of superoxide dismutase and hydroxyproline, but also reduced malondialdehyde content. Due to the limited number of shared differential metabolites exclusively within Kyoto Encyclopedia of Genes and Genomes, comprehensive pathway enrichment analysis was not feasible. Conclusion: This study demonstrates that glabridin and bakuchiol effectively impede photoaging and alleviate skin inflammation in mice. [ABSTRACT FROM AUTHOR]

A 13-year-old male domestic short-hair cat presented for evaluation of labored breathing, hyporexia, and lethargy. Pertinent initial diagnostics yielded leukocytosis, characterized by neutrophilia and monocytosis. Numerous small, round, magenta granules were observed within all neutrophils in Wright-Giemsa-stained blood films on the day of presentation and the day thereafter. No other neutrophil morphologic abnormalities were present, making cytoplasmic toxicity highly unlikely. Hyperadrenocorticism was diagnosed based on the lack of suppression in a low-dose dexamethasone suppression test, and without other diagnostics, the cat was discharged on trilostane therapy. Neutrophil granules did not stain with Alcian blue pH 1.0, periodic acid-Schiff (PAS), PAS and Alcian blue pH 2.5, and toluidine blue. Electron microscopy identified no differences in the morphology of the secretory granules or other neutrophil features. Metabolic screening tests of the cat's urine did not identify a genetic metabolic disorder. However, serum α- and β -hexosaminidase (HexA and HexB) activities were 4.3% and 0% of normal controls, respectively, which is supportive of GM2-gangliosidosis, that is, Sandhoff disorder. However, the historical, clinical, and electron microscopy findings did not provide evidence to confirm this genetic defect. To the author's knowledge, this is the first case of magenta-staining granules within neutrophils in a breed other than a Birman, Siamese, or Himalayan. [ABSTRACT FROM AUTHOR]

We report on the formation of toluidine blue O (TBO) sulfoxide by a self‐sensitized photooxidation of TBO. Here, the photosulfoxidation process was studied by mass spectrometry (MS) and discussed in the context of photodemethylation processes which both contribute to TBO consumption over time. Analysis of solvent effects with D2O, H2O, and CH3CN along with product yields and MS fragmentation patterns provided mechanistic insight into TBO sulfoxide's formation. The formation of TBO sulfoxide is minor and detectable up to 12% after irradiation of 3 h. The photosulfoxidation process is dependent on oxygen wherein instead of a type II (singlet oxygen, 1O2) reaction, a type I reaction involving TBO to reach the TBO sulfoxide is consistent with the results. Density functional theory results point to the formation of the TBO sulfoxide by the oxidation of TBO via transiently formed peroxyl radical or thiadioxirane intermediates. We discover that the TBO photosulfoxidation arises competitively with TBO photodemethylation with the latter leading to formaldehyde formation. [ABSTRACT FROM AUTHOR]

Snakes represent a wide and diverse group of species and have anatomical particularities, such as the renal sexual segment (RSS), a structure located in the kidneys and formed from the hypertrophy of the urinary ducts and nephrons. This study aims at describing the histological aspects of the RSS of Boa constrictor, Epicrates cenchria and Corallus hortulanus, all of which are Brazilian snake species from the Boidae family. The reproductive system and kidneys of five male specimens of E. cenchria, three male specimens of C. hortulanus and two male specimens of B. constrictor were obtained. Tissue samples were processed histologically and different stains used (Toluidine Blue, Alcian Blue and Periodic Acid Schiff). The histological evaluation of the RSS of E. cenchria, C. hortulanus and B. constrictor shows that the RSS in these species varies when comparing individuals in the reproductive period with those which are not. It also allows for the observation of the segment's secretory activity in animals in the reproductive stage (mature sperm in the lumen of the seminiferous tubules) as well as in those which are not. Finally, the histological evaluation also reveals the variation of the secretion product in individuals in the reproductive period, in those which are not, and also among individuals within the same reproductive stage. [ABSTRACT FROM AUTHOR]

Objective: Many dressings are used after the surgical removal of gingival pigmentation. This study aims to compare absorbed dressing (Reso‐Pac®) and eugenol‐free dressing (Coe‐pak) in accelerating wound healing, re‐epithelization and reducing pain after surgical removal of gingival pigmentation using a surgical scalpel. Materials and Methods: This randomized clinical trial (RCT) used a split‐mouth design. According to Hedin's classification, 10 patients with maxillary physiological gingival pigmentation class 3 and 4 were selected. Reso‐Pac® and Coe‐pak were randomly applied on each side after surgery. Landry's scale was used to assess healing index, and re‐epithelization was evaluated using toluidine blue stain. Pain was measured using a visual analogue scale (VAS). Induced pain was evaluated after 1 week by applying airflow on each side for 5 s using VAS. Results: The study group had higher but insignificant differences after 1 and 2 weeks in healing index and re‐epithelization. Pain was insignificantly higher in study group after 2 h but insignificantly higher in the control group in days (1–5). Induced pain was significantly higher (p = 0.015) in the control group after 1 week. Conclusion: Reso‐Pac® can be considered as a viable alternative for Coe‐pak due to its effects in pain reduction and accelerating wound healing. [ABSTRACT FROM AUTHOR]

We report the preparation of a Luffa cylindrica/polypyrrole (LC/PPy) composite and its use for the removal of tartrazine dye (E102) from aqueous media. By use of Fourier transform infrared (FTIR) and energy‐dispersive X‐ray (EDX) spectroscopies, X‐ray diffraction (XRD) and scanning electron microscopy (SEM), we were able to confirm that the PPy chains thoroughly coat the LC matrix, with the composite presenting both amorphous and crystalline phases such as pristine LC. We examined the effects of varying parameters, such as the times allotted for the polymerization and the dye–adsorbent interaction, the E102 initial concentration and the temperature and pH of the solution, on the removal capacity (qe) of the composite. The largest adsorption rate was exhibited by the composite resulting from a 30 min polymerization. At 298 K, the highest value of qe (374.2 mg g−1) occurred at pH 2.0, after 360 min of interaction. The adsorption process occurs spontaneously and is favored by an increase in temperature, with qe = 672.4 mg g−1 at 313 K. We determined that the composite could be used in at least five consecutive adsorption–desorption cycles without noticeable losses of properties. Preliminary experiments indicate that good results were also obtained for the removal of the dyes Remazol Black B, methylene blue and toluidine blue. Overall, these results suggest that the LC/PPy composite is a promising material for use in protocols of removal of dyes from aqueous media. © 2023 Society of Industrial Chemistry. [ABSTRACT FROM AUTHOR]

In this article, the synthesis of a new hybrid compound, candidate as photothermal agent, is proposed, based on TDPP (3,6‐di(thiophene‐2‐yl)‐2,5‐dihydropyrrolo[3,4‐c]pyrrole‐1,4‐dione) and toluidine blue. Electronic structure calculations at the DFT, TD‐DFT and CCSD level of theories were performed to obtain ground and excited states molecular structures, photophysical properties and absorption spectrum of the hybrid and the starting compounds. Additionally, ADMET calculations were performed to predict the pharmacokinetic, metabolic and toxicity properties of the proposed compound. The results showed that the proposed compound is a strong candidate for photothermal agent since (1) it absorbs close to the near‐infrared region, (2) it has low fluorescence and intersystem crossing rate constants, (3) it has accessible conical intersection with low energy barrier, (4) the compound shows lower toxicity than the well know compound toluidine blue, which is used in photodynamic therapy, (5) the compound does not show carcinogenic potential, and (6) it obeys the Lipinski's rule of five, used as a reference for the design of new pharmaceuticals. [ABSTRACT FROM AUTHOR]

Background: The pathogenesis of prurigo nodularis (PN) is considered to be multifactorial, with numerous cells and cytokines confabulating to produce an aberrant immune response. Methods: A cross‐sectional observational study was done in cases of untreated primary prurigo nodularis cases with histopathological assessment in 49 cases from lesional and nonlesional skin with assessment of epidermal and dermal changes, dermal infiltrate, S‐100 and toluidine blue staining to assess the expression of nerve and mast cells. Results: The most common histological changes seen in lesional skin were hyperkeratosis (98%), irregular hyperplasia (69.4%), hypergranulosis (69.4%), subepidermal clefting (6%), vertical collagen bundles (51.0%), and dermal fibrosis (48.9%). Chronic inflammatory infiltrate was seen in all cases (100%) predominantly of lymphocytes (100%) followed by eosinophils (18.4%), plasma cells (8.2%), and neutrophils (2.0%). There was a marked increase in the expression of S‐100 (6.92 ± 3.40 vs. 3.94 ± 2.15, P < 0.001) and toluidine blue (4.99 ± 4.47 vs. 1.22 ± 1.28, P < 0.001) in the lesional skin as compared to the nonlesional skin. Conclusion: We can infer that the epidermal and dermal pathology in PN is related to the infiltrate of lymphocytes, mast cells, and neural hyperplasia which perpetuate the pathogenesis by triggering the itch‐inflammation cycle. Thus, apart from immunosuppressive agents that target lymphocytes and their cytokines, therapy targeted at mast cells and neural proliferation may be needed to treat prurigo nodularis. [ABSTRACT FROM AUTHOR]

Acute ultraviolet B (UVB) irradiation predominantly leads to various skin disorders caused by photodamage. The major causes of UVB‐induced photodamage include oxidative stress, inflammatory infiltration and collagen degradation. The aim of the study was to elucidate whether DNP had protective effect on the skin of KM mice when exposed to UVB irradiation. The DNP protective properties to skin appearance and histopathological alterations in KM mice were evaluated by hematoxylin–eosin staining, toluidine blue staining, Gomori staining and Masson's trichrome staining and mast cell staining. In this study, DNP pretreatment promoted the activities of antioxidant enzymes, including superoxide dismutase, catalase and glutathione peroxidase, while decreased malondialdehyde level in UVB‐irradiated skin, along with downregulation of proteins expression of matrix metalloproteinases and reduction in the level of the proinflammatory cytokines. Based on these findings, we demonstrated that DNP displayed strong ameliorative effects on UVB‐induced acute photodamage for the first time, indicating that it would be a promoting ingredient candidate that could be used in antiphotodamage. [ABSTRACT FROM AUTHOR]

To evaluate the effect of the photodynamic therapy (PDT) using with different photosensitizers on the bond strength of BioRoot RCS (BR) to radicular dentin by comparing with AH Plus (AH). The shaped root canals of 80 human mandibular premolars were divided into four groups based on the final disinfection protocol: NaOCl/EDTA, PDT with methylene Blue (MB), PDT with curcumin (CUR) and PDT with toluidine blue O (TBO) (n = 20). Each group was randomly divided into two subgroups (n = 10), according to the endodontic sealer used for obturation (n = 10). The test of push‐out bond strength (PBS) was performed using an universal test machine. The data were analysed using ANOVA, Tukey post‐hoc, and independent sample t‐test (α = 0.05). MB‐BR group showed the highest PBS at the cervical level (p < 0.05). The selection of the photosensitizer in PDT influenced adhesion of root canal sealers to radicular dentin. [ABSTRACT FROM AUTHOR]

In the present work, we have successfully developed polyaniline supported manganese dioxide (PANI/MnO2) nanocomposite using a facile and inexpensive approach under mild and green conditions by the PANI nanofiber sacrificial oxidation induced reductive conversion of KMnO4 as MnO2. The developed PANI/MnO2 adsorbent material was efficiently utilized for the removal of an organic dye Toluidine Blue (TB) from the aqueous samples.The fabricated adsorbent material was composed of the microstructures that possess grainy and fibrous morphology, and highly uneven surface textures. The FTIR, EDS, and XPS results show the presence of Mn, O, C, and N elements in the nanocomposite. Additionally, from XPS data the survival of the Mn+4 valance state of manganese was recognized. The nano‐thread deposited nano‐fibrous mesoporous nanocomposite holds a high BET surface area (168.096 m2 g−1), and pore volume (0.4428 cm3 g−1).The batch adsorption equilibrium studies showed that 97.51 % of the TB dye was eliminated in 70 min of sorption equilibrium at neutral pH with 92 mg/g of adsorption capacity. The kinetics of the adsorption process is finely portrayed by the pseudo‐second‐order (PSO) kinetic model and the adsorption mechanism is found to be concurrently controlled by intraparticle (IP) and film diffusion (FD) processes. The adsorption isotherm analysis studies revealed that the adsorption of TB dye over the developed PANI/MnO2 adsorbent material occurred with coverage of energetically equivalent binding sites by monolayer formation as substantiated by Langmuir isotherm. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the effectiveness of the original sperm chromatin dispersion (SCD) assay and the toluidine blue (TB) stain to assess DNA fragmentation and chromatin condensation, respectively, in cat sperm obtained by urethral catheterization (CT) and epididymis slicing (EP). CT and EP samples were collected from the same cat, and sperm motility, concentration, morphology, DNA integrity and chromatin condensation were evaluated. As controls, aliquots of the samples were incubated with 0.3 M NaOH and with 1% of dithiothreitol (DTT) to promote DNA fragmentation and chromatin decondensation, respectively. With SCD, four DNA dispersion halo patterns were observed: large, medium, small and no halo. TB staining patterns were as follows: light blue (condensed chromatin), light violet (moderate chromatin decondensation) and dark blue‐violet (high chromatin decondensation). Sperm incubations with NaOH and with DTT were effective in inducing DNA fragmentation and chromatin decondensation, respectively. No significant differences were observed in the percentages of the SCD and TB patterns between samples (CT and EP) and no correlation was observed between sperm head abnormalities and the different SCD and TB patterns. The original SCD technique and the TB stain were adapted to evaluate DNA integrity and chromatin condensation in cat sperm obtained by CT and EP. [ABSTRACT FROM AUTHOR]

Phrenic nerves (PNs) play an important role in respiration; however, very few morphological studies have assessed them. This study aimed to provide control reference values, including the density of large and small myelinated PN fibers, for future pathological studies. We assessed a total of nine nerves from eight cases among consecutive autopsy cases registered to the Brain Bank for Aging Research between 2018 and 2019 (five men and three women, mean age 77.0 ± 7.0 years). The nerves were sampled distally, and their structures were analyzed using semi‐thin sections stained with toluidine blue. The mean and standard deviation of the density of each myelinated fiber of the PN was 6908 ± 1132 fibers/mm2 (total myelinated fiber), 4095 ± 586 fibers/mm2 (large diameter myelinated fiber; diameter ≥7 μm), and 2813 ± 629 fibers/mm2 (small diameter myelinated fiber; diameter <7 μm). There was no correlation between myelinated fiber density and age. This study provides the density measurement of the human PN myelinated fiber, and these findings can be used as reference values for the PN in elderly individuals. [ABSTRACT FROM AUTHOR]

Objectives: The transtibial pull‐out repair (TP) is a relatively new method for treating meniscal root tear; however, the clinical evaluation of its healing effect remains controversial. Due to ethical constraints and limitations of imaging techniques in humans, here we dynamically observe the healing effects of TP and TP with platelet‐rich plasma gel (PRG) at the histological level using an animal model. Methods: Platelet‐rich plasma (PRP) and PRG of rabbits were prepared. Platelet‐derived growth factor (PDGF) and transforming growth factor‐β1 (TGF‐β1) levels in PRP and PRG were determined using an enzyme‐linked immunosorbent assay. A rabbit model of anterior horn tear of the medial meniscus and TP surgery were created. PRG was injected between the anterior horn of the medial meniscus and the tibial tunnel. Rabbits were divided into three groups: the anterior horn tear group (Tear group), the anterior horn tear + TP group (TP group), and the anterior horn tear + TP + PRG group (TP + PRG group). The healing effect was observed dynamically using histopathological studies and biomechanical experiments. Results: The platelet content in PRP significantly increased to approximately 4.57 times that of whole blood. PDGF and TGF‐β1 concentrations in PRG increased to 2.46 and 4.15 times those in PRP, respectively. Hematoxylin and eosin (H&E) and Masson staining showed that the number of inflammatory cells in healing tissue decreased and the collagen fibers significantly increased in TP and TP + PRG groups at 4, 8, and 12 weeks postoperatively compared to those in Tear group. Neatly arranged, interlaced, and dense collagen fibers were found between the anterior horn and bone at 12 weeks. H&E and toluidine blue staining showed that the injury to the femoral condyle cartilage was alleviated. The healing performance in TP + PRG group was better and faster than that in TP group. The maximum tensile fracture strength of the meniscus progressively increased at 8 and 12 weeks postoperatively. Conclusions: Anterior horn injury of the medial meniscus in rabbits can be repaired using the TP technique, and the addition of autologous PRG to the bone tunnel promotes early healing of the meniscus and bone postoperatively. Meanwhile, both treatments can reduce the secondary damage to the cartilage due to osteoarthritis. [ABSTRACT FROM AUTHOR]

Carbon nanotubes (CNTs) are known to be a suitable adsorbent of organic contaminants due to their large surface area and hollow structures; but at the nanoscale, CNTs have poor solubility and can be difficult to remove from solution. Studies have shown that the use of peptides can control the size, shape, and structure of materials during synthesis. In this research, magnetic CNTs were synthesized in a co‐precipitation reaction in the presence of the M6A−B1 peptide to correct these deficiencies in CNTs. Characterization of nanoparticles was facilitated by analytical methods including scanning electron microscopy, infrared spectroscopy, x‐ray diffraction, elemental analysis, and more. Water remediation studies were conducted to study the effect of dye, pH, and adsorbent on the removal of Toluidine Blue O from an aqueous solution. Pseudo‐second order kinetics were used in describing the rate of adsorption while a Langmuir isotherm model was used to study the interaction between the peptide‐functionalized material and the adsorbate. The separation factor, RL, calculated for the studied dye concentrations showed that adsorption with the peptide‐functionalized material was favorable. [ABSTRACT FROM AUTHOR]

The use of doxorubicin (DOX) in clinical practice continues to be challenged by its severe toxicity. DOX cytotoxic activity is not only directed against malignant tumours, given that the treatment will damage healthy tissues as well leading to irreversible injuries. This study aimed to address the in vivo effects of DOX and its co‐administration with a new analog of thioamide; thiocyanoacetamide (TA) on the germinal epithelium. Thus, male rats received either intravenous injection (iv) of 0.03 mg/kg of body weight/week, 0.9% NaCl and were regarded as the control group (CTR), treated with DOX (3.7 mg/kg/week iv), TA [10 mg/kg/day intragastrically (ig)] or a co‐supplementation of DOX and TA. After 50 days, the left testes were dissected and used for toluidine blue, periodic acid‐Schiff (PAS) staining (to evaluate the change in polysaccharides/glycoproteins content), and transmission electron microscopy (TEM) (to assess the morphological damages). To estimate the impact of the test compounds on mitochondrial biogenesis, the expression of NAD‐dependent deacetylase sirtuin‐3 (SIRT‐3) and proliferator‐activated receptor gamma coactivator 1‐alpha (PGC‐1α) were evaluated by immunofluorescence. Apoptotic cells were observed using Hoechst 33324 fluorescent staining. Data showed testicular injuries in the DOX‐treated group, manifested by a significant decrease in total germ cell (GC) number, alteration of Sertoli cell (SC) nucleolus, anchoring junction, along with modifications of the basement membrane (BM) regularity and increase in apoptotic cell count. Mitochondrial aspect and SIRT‐3 and PGC‐1α expression in the testis were unaffected by the DOX. Co‐therapy increased GC number, decreased apoptotic cell count, and restored the BM and anchoring junction regular aspects. This study provides novel insights into understanding DOX‐mediated impairment in rats' testis and might offer some basis for the emerging new alternative therapeutic schemes in male patients undergoing chemotherapy. [ABSTRACT FROM AUTHOR]

To investigate photoactivated disinfection (PAD) to treat acute pseudomembranous stomatitis, an animal model was established. Six‐week‐old male ICR mice were inoculated with Candida albicans under immunosuppression then divided into three groups (15 mice per group). Pseudomembranous area was measured, then mice had 1 mg mL−1 toluidine blue solution spread on the area, left for 1 min (PAD‐1 group) or 2 min (PAD‐2 group), then irradiated with a 750 mW LED red light for 1 min, a control group received no treatment. Fungal load was measured immediately; after 48 h of observation pseudomembranous area and fungal load were measured. The mice were sacrificed and histopathological examination was performed. Before treatment, pseudomembranous area scores were similar (3 to 4 points) in all groups; 48 h after treatment, the treatment groups' scores were lower (1 to 2 points) than the control group (3 to 4 points, P < 0.05). Immediately after treatment and 48 h later, the fungal loads of the treatment groups were lower than the control group (both P < 0.05). Histopathology of the treatment groups improved more than controls. The treatment groups' results were similar. Therefore, this method of PAD, with short treatment time, reduced the fungal load and pseudomembranous area scores in a mouse model of acute pseudomembranous stomatitis and may have clinical potential. [ABSTRACT FROM AUTHOR]

Four genera of the Boidae family are found in Brazil: Eunectes, Corallus, Epicrates, and Boa. Male copulatory organs in snakes are located inside the tail and are called hemipenes. They are double structures in an inverted position that are exposed during copulation. This study describes the macroscopic and histological aspects of hemipenes in Boidae snakes. One Boa constrictor, three Epicrates cenchria, one Corallus hortulanus hemipenes, and one fragment of the muscular retractor penis magnus (m. retractor penis magnus) from Eunectes murinus were included in this study. The structures were evaluated macroscopically and photo‐documented using a stereo microscope. Tissue fragments were included in glycol methacrylate plastic resin, cut into 3‐μm‐thick slices, and stained with 1% toluidine blue for microscopic evaluation. The macroscopic analysis showed that the arrangement and diameter occupied by the m. retractor penis magnus in the hemipenis and the internal and external cavernous bodies differ between species. The histological analysis showed that the hemipenis of B. constrictor, E. cenchria, and C. hortulanus has stratified epithelium in the external portion supported by dense connective tissue with blood vessels and muscle tissue. Although the literature reports the presence of skeletal musculature in the hemipenis, striated musculature with a centrally located nucleus in the fibre bundle, typical of cardiac musculature in mammals, was observed in the studied species. [ABSTRACT FROM AUTHOR]

Background: Macrophages are the only inflammatory cells that can penetrate the closed nucleus pulposus and their polarization plays an important role in intervertebral disc degeneration (IVDD). This paper attempted to investigate the pathogenesis of IVDD by altering the polarization state of macrophages. Methods: Macrophage RAW264.7 cells were induced by interferonγ (IFN‐γ) and lipopolysaccharide (LPS). The polarization of RAW264.7 cells was estimated by western blot and immunofluorescence. The expressions of inflammatory factors were detected by ELISA. Subsequently, RAW264.7 cells were treated with different concentrations of minocycline (Mino) and sinomenine (Sino), followed by the assessment of cell viability with cell counting kit‐8 kit. Then, RAW264.7 cell culture medium was collected for the culture of human nucleus pulposus cells (NPCs). Toluidine blue staining and type II collagen staining were applied to assay the level of type II collagen. The cell apoptosis, oxidative stress, and nitric oxide (NO) level were appraised by TUNEL, oxidative stress kits and NO kit, respectively. Western blot was employed to test the levels of apoptosis‐ and oxidative stress‐related proteins. Results: IFN‐γ and LPS could induce M1 polarization of RAW264.7 cells. Mino and Sino could reduce the polarization of RAW264.7 cells toward M1. M1‐polarized medium inhibited LPS‐induced activity, inflammation, and damage of NPCs, which were enhanced by Mino and Sino in medium. Conclusion: M1 polarization of macrophages promoted LPS‐induced inflammation and damage of NPCs. [ABSTRACT FROM AUTHOR]

Efficient pharmacotherapy of cancer is related to accurate recognition of genetic mutations and epigenetic alterations in the early‐stage diagnosis. In the present study, a novel optical genosensor based on toluidine blue as photonic probe was developed to detection of DNA methylation using hybridization of pDNA with cDNA. Biomedical analysis was performed using UV‐vis and fluorometric methods. For the first time, this strategy was applied for the distinction of methylated DNA from unmethylated‐DNA‐based on the interaction of optical probe with methylated‐DNA and unmethylated DNA. Fluorescence spectroscopic data showed that poly‐toluidine blue could be bind to DNA sequences and lead to different fluorescence patterns and could be used as an efficient geno‐platform for the sensitive bioassay of mutation. The excitation and emission wavelengths were 580 and 630 nm, respectively. Non‐binding of mismatch sequences with the optical probe was used as negative control. Under optimal conditions, linear range was 1 zM to 0.2 pm and the lower limit of quantitation was obtained as target concentrations ranging 1 zM. The designed genosensor showed high capability to distinct methylation from un‐methylated. Therefore, the designed DNA‐based bioassay could detect DNA methylation significantly. Finally, bioanalysis of real samples showed that the designed genosensor could use to detect DNA methylation which is a new platform for point of care analysis. [ABSTRACT FROM AUTHOR]

A 14‐year‐old, female, spayed warren hound presented with an acute history of collapse, pale mucous membranes and abdominal distension. Abdominal ultrasound and abdominocentesis revealed a haemoabdomen secondary to a ruptured splenic mass. The dog underwent an emergency splenectomy and splenic histopathology, including toluidine blue staining, was consistent with a mast cell tumour. The dog was diagnosed with a primary splenic mast cell tumour based on the absence of a primary mast cell tumour in another location at the time of presentation or in a review of the dog's medical record. Following surgery, staging identified hepatic infiltration, and adjuvant chemotherapy with vinblastine and prednisolone was started. The dog was euthanased 97 days after surgery due to progressive disease and recurrence of the haemoabdomen. Visceral mast cell tumours should be considered as an uncommon differential diagnosis in dogs presenting with haemoabdomen due to a ruptured splenic mass. [ABSTRACT FROM AUTHOR]

Background: Photodynamic therapy is a two‐step procedure, involving the use of photosensitizing agents followed by selective illumination of the target lesion with visible light. Photodynamic therapy has been described recently as a promising strategy for treatment of leishmaniasis. This study aims to evaluate the in vitro phototoxic, morphological, and apoptotic effect of methylene blue, toluidine blue, chloro‐aluminum phthalocyanine, and pheophorbide a‐mediated photodynamic therapy on the viability of Leishmania tropica promastigotes. Methods: Parasites were treated with methylene blue, toluidine blue, chloro‐aluminum phthalocyanine, and pheophorbide a or/and methylene blue, toluidine blue, chloro‐aluminum phthalocyanine, and pheophorbide a‐mediated photodynamic therapy, and cell proliferation, morphological changes, and apoptosis were evaluated by XTT, giemsa staining, DAPI staining, and DNA fragmentation, respectively. Results: Parasite viability was significantly different in between the groups treated with methylene blue, toluidine blue, and pheophorbide a, with or without irradiation. chloro‐aluminum phthalocyanine treatment did not lead to any alterations in cell viability in Leishmania tropica promastigotes with or without irradiation. DAPI staining results indicated that apoptotic bodies and nucleus fragmentation started to be visible in methylene blue, chloro‐aluminum phthalocyanine, and pheophorbide a‐mediated photodynamic therapy groups. DNA ladder pattern which is used to define apoptosis was observed in irradiated methylene blue, chloro‐aluminum phthalocyanine, and pheophorbide a groups. Conclusions: The results revealed that apoptosis‐induced cell death was observed in Leishmania tropica promastigotes after the application of photosensitizers in combination with light irradiation. [ABSTRACT FROM AUTHOR]

Amyloid precursor‐like protein‐2 (APLP2) and its C‐terminal fragments (CTFs) are expressed at high levels in pancreatic cancer cells and knockdown of APLP2 expression inhibits tumor growth. CTFs are released from APLP2 by beta‐secretase (BACE). In this study, our goal was to determine whether methylene blue (MethB) and toluidine blue O (TBO) could be used to slow down the growth and viability of pancreatic cancer cells (Hs 766T). We found that TBO and MethB decreased the growth and viability of Hs 766T cells in a dose‐ and time‐dependent manner compared to vehicle‐treated control, as demonstrated by MTT and trypan blue exclusion assays. Although TBO led to decreased expression of APLP2, MethB did not show any significant effect on APLP2. However, both MethB and TBO reduced BACE activity and the levels of APLP2 CTFs in Hs 766T cells. In conclusion, MethB and TBO may be valuable candidates for the treatment of pancreatic cancer by targeting APLP2 processing. [ABSTRACT FROM AUTHOR]

Drought stress is a major abiotic stress that limits rice yield. Therefore, the development of new varieties tolerant to drought stress is a high priority in breeding programs. In this study, 150 rice M10 mutant lines, previously developed using gamma‐ray irradiation, were used, and a drought‐insensitive rice mutant (ditl1) was selected by drought stress screening. The ditl1 mutant exhibited significantly decreased water loss, leaf curling, and H2O2 accumulation under drought stress. Chlorophyll leaching assay and toluidine blue staining suggested lower cuticle permeability in ditl1 mutants than in wild‐type (WT) plants. In addition, transmission electron microscopy revealed that ditl1 plants accumulated more cuticular wax on the epidermal surface. Whole‐genome resequencing analysis suggested that the deletion of a single nucleotide on the LOC_Os05g48260 gene, a putative ortholog of WSD1 (wax ester synthase/diacylglycerol O‐acyltransferase in Arabidopsis), maybe be the gene responsible for the drought insensitive phenotype of ditl1. The ditl1 mutant will be a valuable breeding resource for developing drought stress tolerant rice cultivar. [ABSTRACT FROM AUTHOR]

Mast cells, known as pro‐inflammatory effector cells, are immunocytes present in the meninges and may be involved in the pathophysiology of migraine. This study aims to evaluate the histomorphometric parameters of mast cells located in the convexity of the human intracranial dura mater. For this, samples of intracranial dura mater from eight human fresh cadavers were collected between 8‐ and 24‐h post‐mortem. The whole samples were fixed and, subsequently, two fragments of 1.5 cm² each were cut from four different areas of the dura mater convexity, containing a segment of the middle meningeal artery, totaling 64 fragments. After histological processing, the fragments were submitted to microtomy (5 and 10 µm), stained with toluidine blue (0.1%), or immunohistochemically labeled for tryptase, and analyzed using optical microscopy. The following histomorphometric parameters were evaluated: distance from mast cells to vessels, the density of mast cells, and percentage of mast cells with degranulation. Histomorphometric analyzes showed a higher density of mast cells in the vicinity of blood vessels (arterial and venous), with distances around 0–150 µm. A greater number of mast cells was detected near venous vessels in the periosteal layer (17.0 ± 10.1 cells/mm²) than in the meningeal layer (14.1 ± 7.0 cells/mm²) (p < 0.05). Mast cells from the region close to the superior sagittal sinus were found in greater quantity close to the venous vessels (16.7 ± 10.1 cells/mm²) than to the arterial vessels (11.2 ± 7.5 cells/mm²) (p < 0.05). In short, in the convexity of the human intracranial dura mater, mast cells are located close to blood vessels, with a greater number of cells next to the venous vessels of the periosteal layer and in the proximal region of the superior sagittal sinus. [ABSTRACT FROM AUTHOR]

Meniscus allograft transplantations (MATs) represent established surgical procedures with proven outcomes. Yet, storage as frozen specimens and limited cellular repopulation may impair graft viability. This proof‐of‐concept study tests the feasibility of injecting allogeneic mesenchymal stromal/stem cells (MSCs) in meniscus allograft tissue. We investigated the injectable cell quantity, survival rate, migration, and proliferation ability of MSCs up to 28 days of incubation. In this controlled laboratory study, seven fresh‐frozen human allografts were injected with human allogeneic MSCs. Cells were labeled and histological characteristics were microscopically imaged up to 28 days. Mock‐injected menisci were included as negative controls in each experiment. Toluidine blue staining demonstrated that a 100‐µl volume can be injected while retracting and rotating the inserted needle. Immediately after injection, labeled MSCs were distributed throughout the injection channel and eventually migrated into the surrounding tissues. Histological assessment revealed that MSCs cluster in disc‐like shapes, parallel to the intrinsic lamination of the meniscus and around the vascular network. Quantification showed that more than 60% of cells were present in horizontally injected grafts and more than 30% were observed in vertically injected samples. On Day 14, cells adopted a spindle‐shaped morphology and exhibited proliferative and migratory behaviors. On Day 28, live/dead ratio assessment revealed an approximately 80% cell survival. The study demonstrated the feasibility of injecting doses of MSCs (>0.1 million) in meniscus allograft tissue with active cell proliferation, migration, and robust cell survival. [ABSTRACT FROM AUTHOR]

Sperm cryopreservation as a routine technique in assisted reproductive technique (ART) laboratories has detrimental effects on spermatozoa. Various methods have been introduced to improve it. The aim of this research was to evaluate the effects of L‐proline supplementation in cryopreservation medium on normozoospermic semen samples. A total of 30 semen samples were collected from normozoospermic men. Cryopreservation media were supplemented with different concentrations of L‐proline (0, 1, 2 and 4 mmol/L). The semen samples were cryopreserved. After thawing, sperm parameters and chromatin integrity (aniline blue (AB), toluidine blue (TB), sperm chromatin dispersion test (SCD) and chromomycin A3 (CMA3)), reactive oxygen species (ROS), and total antioxidant capacity (TAC) and malondialdehyde (MDA) levels were evaluated. A total of 4 mmol/L L‐proline significantly improved progressive motility and viability (p < 0.05). MDA and ROS levels significantly diminished in samples were cryopreserved by 4 mmol/L L‐proline supplemented cryopreservation media (p < 0.001). Also, it significantly increased TAC level. Also, chromatin damages (AB, TB and CMA3) significantly improved in samples were cryopreserved by 4 mmol/L L‐proline supplemented cryopreservation media (p < 0.05). The results support that the usage of L‐proline supplemented cryopreservation media to improve sperm quality after cryopreservation. [ABSTRACT FROM AUTHOR]

Purpose: The aim of this study was to investigate the presence of synovial mast cells (MCs) in hip and knee tissue from osteoarthritis (OA) patients and to correlate them with clinical and radiological data. Methods: Synovial tissue was obtained during arthroplasty from 60 patients, 30 with knee OA and 30 with hip OA. Control synovial tissue was obtained from 30 patients without OA, 15 undergoing above-knee amputation and 15 receiving a hip replacement for fracture. Before surgery, the radiographic findings were graded according to the Kellgren-Lawrence system and clinical data including pain (VAS) and functional information (KOOS and HOOS) was collected. The tissue was stained with hematoxylin–eosin and toluidine blue for histochemistry and incubated with CD117 and CD31 antibodies for immunohistochemistry. MC and vessel number and synovitis score were determined in all samples. Results: Mean MC number, synovitis score and vessel number were significantly higher in the OA samples (p < 0.05) than in control tissue. MC number correlated with the synovitis score and disease severity in both patient groups. Conclusions: The prevalence of MCs in synovium from OA patients and their association with synovial inflammation and pain suggest a role for them in OA pathophysiology. [ABSTRACT FROM AUTHOR]

This research demonstrates the value of laser scanning confocal microscopy (LSCM) as a research tool in osteological studies, and diagenetic studies in particular. LSCM combines properties of light and scanning electron microscopy using laser light to excite fluorophores throughout the z‐axis, developing a 3‐D image. Using differential staining and selecting for specific wavelengths of light, one can image targeted materials. This research is divided into two parts: visualizing bone structures such as proteins and their decompositional products and visualizing diagenesis. Part one of this study utilized pig bones as a means of testing the overall ability of LSCM to fluoresce bone. Twenty‐three samples were imaged, including 13 samples from a decompositional study conducted 5 years previous, and 10 "fresh" samples collected from a commercial butcher. This part of the study determined that protein and organic components of the bone could be fluoresced and diagenetic alteration could be imaged. The second part of the study used human samples as a means of imaging and mapping diagenetic alterations. The second part of the study used 13 samples, including 4 clinical, 7 ancient, and 2 modern controls. The pig study used Basic Fuchsin and SlowFade Gold stains, while the human study used toluidine blue. Images were also taken with unstained elements. The results of the non‐human study found that a fresh bone fluoresced differently than that of a 5‐year subset, while the results of the human study confirmed these findings and determined that the bone diagenesis can be mapped using LSCM. [ABSTRACT FROM AUTHOR]

Nanoporous of poly (vinylpyrrolidone)/Fe3O4@SiO2 has been synthesized using ultrasonic‐assisted‐gamma irradiation technique and characterized by transmission electron microscopy (TEM), scanning electron microscope (SEM)/energy‐dispersive spectroscopy (EDX), Fourier transform infrared (FTIR), X‐ray diffraction (XRD), Vibrating Sample Magnetometer (VSM), and Brunauer–Emmett–Teller (BET). The efficiency evaluation of PVP/Fe3O4@SiO2 nanoporous catalyst for degradation of toluidine blue (TB) by sono‐photo‐Fenton (US/UV/FE) processes has been investigated using modified reactor under the influence of magnetic field (MF) 250 Gauss. PVP molecules prevent ions leaching and render Fe3O4@SiO2 nanoparticles to bind with pollutant molecules. Comparative effect of sonication assists on photo‐Fenton processes has been studied. The TB degradation was examined at different pHs and H2O2 concentration. After 60 min, the photo‐catalytic degradation rate of TB is attained to optimum value (83%) at pH 4. The photo‐Fenton catalytic degradation rate of TB decreased (36.39%, 31.84%, and 23.92%) within (0.5, 1.0, and 1.5 M) of H2O2, respectively. The degradation of TB by sono‐photo‐Fenton process was 76.55 and 95.3% after 60 and 10 min in absence and presence of MF, respectively. Negligible amounts of Fe leaching from PVP/Fe3O4@SiO2 confirm high stability in oxidation systems. BET surface area and pore volume of PVP/Fe3O4@SiO2 were 43.50 m2 g−1 and 0.1798 cm3 g−1, respectively, with pore size distribution of 16.53 nm indicating their nanoporosity structure. Herein, the novelty is evaluated through the effect of MF and ultrasound‐assisted photo‐Fenton system for TB degradation using novel PVP/Fe3O4@SiO2 nanoporous catalyst that easily recovered from the reaction solution by external magnet. The PVP/Fe3O4@SiO2/US/UV/FE/MF hybrid system can be introduced as a successful advanced treatment process for degradation of dyes from wastewater with high degradation efficiency in short time. [ABSTRACT FROM AUTHOR]

Morphofunctional characteristics of the large intestine are rarely explored to understand the physiology, behavior and ecology of neotropical primates. In this study, we analyzed the histometric parameters of the large intestine of hybrid marmosets (Callithrix sp.) captured in forest fragments of Viçosa‐Brazil, under seasonal interference. These animals were predominantly insectivorous in the rainy season and gummivores in the dry season. Large intestine fragments were collected and processed according to histological methods and stained with toluidine blue for general analysis, periodic acid of Schiff (PAS) and Alcian blue (AB) for goblet cells, Grimelius and Masson Fontana for argyrophil and argentaffin endocrine cells. Several histometric parameters were more expressive in the large intestine of the rainy season marmosets: greater thickness of the parietal layers, greater number of argyrophil and argentaffin endocrine cells, and AB‐positive goblet cells, characteristics favor secretomotor functions and reduce the passage time of the fecal bolus, which is consistent with an insectivorous diet. In contrast, parameters such as crypt width, height of the absorptive cells and striated border, and the number of PAS‐positive cells were more expressive in the dry season marmosets, reflecting the need for longer passage time for digestion and absorption of food items from tree gum, which are more complex and demand the action of microorganisms present in the large intestine, as well as greater protection against the abrasive action of dietary fibers and against microorganisms. Thus, it can be said that the marmoset's large intestine has morphological adaptations to maximize energy intake from the diet, which alternates under the influence of seasonality. [ABSTRACT FROM AUTHOR]

Previous study has suggested the colonic nontoxicity and obesity inhibition of food‐grade κ‐carrageenan in obese mice. Further study using transcriptome is important to provide further understanding on the gene expressions of inflammation and obesity. Here, the obese mice without any treatment (HFD) or with 5% food‐grade κ‐carrageenan diet intervention (H5%) were used to perform colonic transcriptome sequencing. The results showed that genes involved in the inflammatory pathways or tight junction protein encoding were not significantly dysregulated by 5% carrageenan. However, the expression of lipid metabolism genes meaningfully changed as evidenced by the decreased gene levels of adipocytokines, lipogenesis, lipid absorption and transport, and the increased adipolysis and oxidation. In addition, the carrageenan metabolism experiments by toluidine blue (TB) staining of colon and high‐performance size exclusion chromatography (HPSEC) of feces supernatant showed that the food‐grade κ‐carrageenan was not absorbed or significantly degraded in the digestive tract of obese mice. Hence, the fact that food‐grade κ‐carrageenan was not significantly metabolized by the organism and did not cause obvious dysregulation of colonic inflammatory genes provided evidences for its noncolonic toxicity in obese mice. An anti‐obesity potential of food‐grade κ‐carrageenan was probably mediated by the regulation of lipids metabolism‐related genes. [ABSTRACT FROM AUTHOR]

Mutualistic mycorrhiza in orchids: evidence from plant-fungus carbon and nitrogen transfers in the green-leaved terrestrial orchid Goodyera repens. I sikokianus i and albino mutants of so-called rhizoctonia-associated orchids, including I Goodyera velutina i (8.8 ± 0.9‰; Suetsugu I et al i ., 2019) and I Cypripedium i I debile i (9.8 ± 0.5‰), are similar to those of mycoheterotrophic orchids exploiting saprotrophic nonrhizoctonia fungi. Keywords: carbon dynamics; mixotrophy; mycorrhizas; partial mycoheterotrophy; radiocarbon dating; rhizoctonias; saprotrophic fungi; Sebacinales EN carbon dynamics mixotrophy mycorrhizas partial mycoheterotrophy radiocarbon dating rhizoctonias saprotrophic fungi Sebacinales 1670 1675 6 08/07/21 20210901 NES 210901 Controversy on the nutritional mode of rhizoctonia-associated orchids Mycorrhizas usually engage in mutualism, in which soil fungi contribute minerals to plants that in turn supply photosynthetically fixed carbon (C) to their fungal partners (Smith & Read, 2008). Although all orchids depend on symbiotic fungi for essential nutrients (including C) during their initial development (Dearnaley I et al i ., 2016), most orchid species exhibit some level of autotrophy at the adult stage. [Extracted from the article]

In this study, a novel biosensor based on molecular imprinting polymer (MIP) methodology was fabricated toward recognition of carcinoembryonic antigen (CEA). For this purpose, poly (toluidine blue) (PTB) was electropolymerized on the surface of gold electrode in the absence and presence of CEA. So, the target molecules were entrapped into the imprinted specific cavities of MIP. Obtained results show that, the binding affinity of the MIP system was significantly higher than that of revealed for the nonimprinted polymer (NIP) system, MIP‐based biosensor revealed linear response from (0.005 to 75 μg/L) and low limit of quantification of (0.005 μg/L) by using chronoamperometry technique, leading to CEA monitoring in real and clinical samples. Thus, a novel technique for rapid, simple, sensitive and affordable monitoring of CEA (LLOQ = 0.005 μg/L) has provided through developed biosensor. From a future perspective, moreover, this method can be considered as an applicable candidate in biomedical and clinical analysis for point‐of‐care usages. [ABSTRACT FROM AUTHOR]

Among several oil/water emulsion separation technologies, the utilization of nanoparticle‐decorated membranes with diverse functionalities has received considerable attention in recent years, particularly if the antifouling capacity can be improved. In this article, we propose a new membrane based on surface‐hydrolyzed polyacrylonitrile electrospun membranes and/or decorated with amine‐functionalized Si‐MCM‐41 nanoparticles to be used as oil/water emulsion separation treatment and to determine their antifouling ability. X‐Ray photoelectron spectrometry, attenuated total reflectance Fourier transform infrared spectroscopy, and toluidine blue O assay, scanning electron microscopy, contact angle measurements for oil under water and thermogravimetry were used for characterizing the membranes and an assay of permeability was developed to quantify the diffusion of oil molecules across the electrospun membrane. The electrospun and/or decorated membranes showed an underwater oleophobic wettability, which can separate oil‐in‐water emulsions with 87% separation efficiency, results of fouling experiments, evaluated in terms of rejection and flux recovery ratio, exhibited good antifouling ability, but the membrane decoration process did not lead to superior outcomes compared with undecorated membranes. [ABSTRACT FROM AUTHOR]

Tea, which is usually made from the new shoots of tea plants (Camellia sinensis), is one of the most popular and healthy beverages. The tenderness of new shoots plays a critical role in the production of high‐quality tea; however, the factors affecting the tenderness of the new shoots of tea have not been extensively studied. Here, we show that cellulose accumulation is negatively correlated with the tenderness of new shoots, including leaves and stems, by characterizing the cellulose content and visualizing cellulose in the new shoots of two cultivars ("Huangjinya" and "Yujinxiang") via toluidine blue staining. We further found that the number of cells with secondary cell walls (SCWs) and the thickness of SCWs increased with the maturity degree of stems in both cultivars. Because cellulose is the most abundant polymer in SCW, we next identified three cellulose synthase CsCesAs, CsCesA4, 7, and 8A, through homologous alignment with Arabidopsis AtCesAs. Protein sequence analysis revealed that these proteins were closely related based on the phylogenetic analysis. We finally showed that the gene expression of the three CsCesAs was positively correlated with the maturity degree and cellulose content of the new shoots. Thus, our data suggest that cellulose accumulation, especially in the SCWs, is negatively correlated with the tenderness of new shoots in tea plants. [ABSTRACT FROM AUTHOR]

Objective: Intraoperative crush/squash smear or frozen section consultation is routinely performed at several centers and offers rapid onsite assessment of tumor type and provides invaluable information to the neurosurgeons. The WHO classification of central nervous system neoplasms underwent a paradigm shift in 2016 with the incorporation of molecular data with the morphological features, such that several new entities came to be distinctly defined. With this paper, we present our experience at intraoperative consultation of brain tumors arising in posterior fossa and aim to apprise the pathologists with the spectrum of cytomorphologic appearances that can occur during such consultation and highlight the diagnostic dilemmas and pitfalls encountered in this setting. Methods: This is a retrospective observational study illustrating the salient morphological features of commonly encountered brain tumors arising in the posterior fossa (prototype example of each type) reported at our institute over a period of seven years. Both squash smears and rapid snap frozen section were prepared and stained with Toluidine blue and rapid hematoxylin and eosin (H&E) stains. Results and conclusions: While the majority of tumors at this location comprise of pilocytic astrocytoma, ependymoma, and medulloblastoma, some rare examples may also arise; common differentials must be considered and prudently excluded to arrive at the diagnosis which is crucial in guiding the neurosurgeon. Both squash smears and rapid frozen section should be prepared and complement each other for rapid on‐site evaluation. Intraoperative diagnosis has long played an important role in guiding neurosurgical biopsies and resections, and a high degree of accuracy in such rapid diagnoses is the norm. Awareness about the morphological features in combination with radiological features and the location of the lesion helps in clinching the correct diagnosis. [ABSTRACT FROM AUTHOR]

In a previous study, we reported that ethylene treatment facilitated boil‐peeling in persimmons and in several other fruits; however, the mechanism underlying the facilitating effect of ethylene was not examined in detail. Thus, in this study, we investigated the effect of ethylene treatment on the peel characteristics of persimmons, that facilitated boil‐peeling, using chemical, genomic, and histochemistry analyses. The results of the study showed that the ethylene‐related genes, DK‐ACS1 and DK‐ACO2, and the pectinase‐active gene DKPG were not expressed, even though a minor increase in ethylene generation was observed after ethylene treatment. Conversely, significant accumulation of toluidine blue O and ruthenium red dyes were observed in the sarcocarp and exocarp of the fruits, indicating an increase in the quantity of polysaccharides, including pectic substances, at the site. The results also indicate that the increased cellulase activity observed in the pericarp of the fruits may be due to the aging of the fruits, and not necessarily as a result of ethylene treatment. Furthermore, ethylene treatment increased the quantity of polysaccharides, including pectic substances, directly below the pericarp, which caused the dissolution of the site, resulting in peeling. This study provides new insights on the effect of ethylene on boil‐peeling in persimmons and provides a foundation for future research studying the effect of heat treatment in the peeling of fruits or tomato. [ABSTRACT FROM AUTHOR]

This study focuses on the deposition of plasma polymer films (PPFs) on hydrogels that could act as permeation barriers for aqueous species. The deposited PPFs on hydrogels were studied in terms of their ability to resist the permeation of an aqueous dye molecule after 24‐h exposure to an aqueous solution and mechanical stress. Permeation of the dye toluidine blue O through the hydrogels after plasma polymerisation was at least 75% less than that through an untreated hydrogel. The X‐ray photoelectron spectroscopy analysis showed introduction of O‐ and N‐rich chemical functionalities in the surface chemical composition of the hydrogel after plasma polymerisation. Scanning electron microscope imaging pointed out that milder plasma conditions result in uniform films that do not degrade upon exposure to water or mechanical stress. [ABSTRACT FROM AUTHOR]

In vitro experiments confirmed that antibacterial photodynamic treatment (aPDT) inactivates periodontal pathogens. However, more effective sterilization is needed in the complex oral environment. This study tested whether dihydroartemisinin (DHA) enhanced the photokilling effect of aPDT on Porphyromonas gingivalis (P. gingivalis) in planktonic and biofilm states. aPDT combining toluidine blue O (TBO) with 630 nm red light was performed on bacterial suspensions and biofilms in vitro with different final concentrations of DHA (10, 20 and 40 μg mL−1). The sensitization mechanism was preliminarily investigated by uptake experiments. The above experiments were repeated with different incubation times (30, 60, 120 s). Porphyromonas gingivalis biofilms exhibited significantly higher resistance to aPDT than P. gingivalis in suspension under the same experimental parameters. DHA alone had no cytotoxic effect on P. gingivalis with or without light irradiation. In either bacterial suspensions or biofilms, DHA concentration‐dependently enhanced the photokilling effect of aPDT and increased TBO uptake by P. gingivalis. Prolonged incubation time enhanced the photokilling efficiency of aPDT until cellular TBO uptake reached saturation. DHA can enhance aPDT activity against P. gingivalis in planktonic and biofilm states. DHA also accelerated TBO uptake, reducing incubation time. [ABSTRACT FROM AUTHOR]

Toluidine blue O (TBO) is a water‐soluble photosensitizer that has been used in photodynamic antimicrobial and anticancer treatments, but suffers from limited solubility in hydrophobic media. In an effort to incrementally increase TBO's hydrophobicity, we describe the synthesis of hexanoic (TBOC6) and myristic (TBOC14) fatty acid derivatives of TBO formed in low to moderate percent yields by condensation with the free amine site. Covalently linking 6 and 14 carbon chains led to modifications of not only TBO's solubility, but also its photophysical and photochemical properties. TBOC6 and TBOC14 derivatives were more soluble in organic solvents and showed hypsochromic shifts in their absorption and emission bands. The solubility in phosphate buffer solution was low for both TBOC6 and TBOC14, but unexpectedly slightly greater in the latter. Both TBOC6 and TBOC14 showed decreased triplet excited‐state lifetimes and singlet oxygen quantum yields in acetonitrile, which was attributed to heightened aggregation of these conjugates particularly at high concentrations due to the hydrophobic "tails." While in diluted aqueous buffer solution, indirect measurements showed similar efficiency in singlet oxygen generation for TBOC14 compared to TBO. This work demonstrates a facile synthesis of fatty acid TBO derivatives leading to amphiphilic compounds with a delocalized cationic "head" group and hydrophobic "tails" for potential to accumulate into biological membranes or membrane/aqueous interfaces in PDT applications. [ABSTRACT FROM AUTHOR]

Aims: The accuracy of toluidine blue (TB) and chemiluminescence for diagnosing oral cancer and pre‐cancer was evaluated. Methods: Two authors (working independently) comprehensively reviewed six databases (PubMed, Cochrane database, Embase, Web of Science, SCOPUS and Google Scholar) from their dates of inception until March 2020. Oral mucosal disorder, as detected by TB, was compared with that detected by chemiluminescence. True‐positive, true‐negative, false‐positive and false‐negative data were extracted for each study. Methodological quality was evaluated using the Quality Assessment of Diagnostic Accuracy Studies tool (ver. 2). The extent of interrater agreement was also assessed. Results: Twenty‐nine prospective and retrospective studies were included. The diagnostic odds ratio (DOR) of TB was 7.017 (95% confidence interval [CI], 4.544; 10.836). The area under the summary receiver operating characteristic curve was 0.766. The correlation between the sensitivity and the false‐positive rate was 0.196, indicating the absence of heterogeneity. TB exhibited moderate interrater reliability (0.6777; 95% CI, 0.43; 0.7455). Compared with chemiluminescence, as used in nine studies, TB had a lower sensitivity (0.659 vs 0.841), but a higher specificity (0.809 vs 0.345), negative predictive value (0.766 vs 0.690) and DOR (10.565 vs 5.203). Compared with clinical examination, as used in four studies, TB method had a higher sensitivity (0.891 vs 0.891), specificity (0.739 vs 0.634), negative predictive value (0.920 vs 0.714) and DOR (28.491 vs 8.526). Subgroup analysis showed that screening for severe dysplasia or more severe disease was significantly more sensitive, but less specific, than screening for all dysplasias. Conclusions: Although the diagnostic accuracy of TB in the diagnostic work‐up of oral cancer and pre‐cancer was higher than that of clinical examination, it was not high enough for TB to reliably be used alone. Instead, it should be combined with chemiluminescence or other diagnostic tools. [ABSTRACT FROM AUTHOR]

Toluidine blue O (TBO) is a water‐soluble photosensitizer that has been used in photodynamic antimicrobial and anticancer treatments, but suffers from limited solubility in hydrophobic media. In an effort to incrementally increase TBO's hydrophobicity, we describe the synthesis of hexanoic (TBOC6) and myristic (TBOC14) fatty acid derivatives of TBO formed in low to moderate percent yields by condensation with the free amine site. Covalently linking 6 and 14 carbon chains led to modifications of not only TBO's solubility, but also its photophysical and photochemical properties. TBOC6 and TBOC14 derivatives were more soluble in organic solvents and showed hypsochromic shifts in their absorption and emission bands. The solubility in phosphate buffer solution was low for both TBOC6 and TBOC14, but unexpectedly slightly greater in the latter. Both TBOC6 and TBOC14 showed decreased triplet excited‐state lifetimes and singlet oxygen quantum yields in acetonitrile, which was attributed to heightened aggregation of these conjugates particularly at high concentrations due to the hydrophobic "tails." While in diluted aqueous buffer solution, indirect measurements showed similar efficiency in singlet oxygen generation for TBOC14 compared to TBO. This work demonstrates a facile synthesis of fatty acid TBO derivatives leading to amphiphilic compounds with a delocalized cationic "head" group and hydrophobic "tails" for potential to accumulate into biological membranes or membrane/aqueous interfaces in PDT applications. [ABSTRACT FROM AUTHOR]

Quercetin, an antioxidant derived from plants, can play a beneficial role in the protection of various tissues against ischemia‐reperfusion injuries (IRI). The purpose of the present research was to investigate the protective effects of quercetin on gastrocnemius muscle ischemia‐reperfusion. A total of 80 adult male Wistar rats (weights: 250–300 g) were divided into ten groups (n = 8 per group). We used silk 6.0 surgical thread to create a knit to occlude the femoral artery and vein for 3 hr. The treated groups, which comprised half of each experimental group, received intraperitoneal injections of 150 mg/kg quercetin after the ischemia. Blood flow was subsequently reestablished in the reperfusion phase. The rats were kept in reperfusion for 3, 7, 14, or 28 days after which they were killed with high doses of anesthetic drugs, and the gastrocnemius muscles were removed and fixed. Tissue processing, hematoxylin and eosin and toluidine blue staining, and immunohistochemistry were used to assess tumor necrosis factor‐α (TNF‐α) and nuclear factor κB (NF‐κB) levels. A comparison between treated and untreated ischemic sites showed that on the third day of reperfusion, the severity of edema and NF‐κB level decreased significantly; on the 7th day of reperfusion, the severity of edema and the levels of TNF‐α and NF‐κB decreased significantly; and on the 14th day of reperfusion, all of the parameters showed significant decreases. On the 28th day of reperfusion, there were significantly decreased levels of TNF‐α and NF‐κB, and decreased mast cell infiltration when compared with the untreated groups. According to the results, administration of quercetin after ischemia could significantly prevent gastrocnemius muscle IRI. [ABSTRACT FROM AUTHOR]

The aim is to evaluate the accuracy of autofluorescence for screening oral cancer in comparison with toluidine blue staining and clinical examination. Oral mucosal disorders detected by autofluorescence were compared with those detected using toluidine blue staining. Methodological quality was evaluated using the Quality Assessment of Diagnostic Accuracy Studies tool. The diagnostic odds ratio (DOR) for autofluorescence was 8.197 (95% confidence interval, 4.018‐16.723). The area under the summary receiver operating characteristic curve (AUC) was 0.815. Compared with toluidine blue, autofluorescence had a similar sensitivity, negative predictive value, and AUC but a lower specificity and DOR. Compared with clinical examination, the autofluorescence had a higher sensitivity but lower specificity, DOR, and AUC. Although the diagnostic accuracy of autofluorescence in the screening work‐up of oral cancer and precancer was more sensitive than those of clinical examination and toluidine blue staining, it was not accurate enough to be used alone reliably. [ABSTRACT FROM AUTHOR]