Search

Exposure of bacteria to low concentrations of biocides can facilitate horizontal gene transfer, which may lead to bacterial adaptive responses and resistance to antimicrobial agents. The emergence of antibacterial resistance not only poses a significant concern to the dairy industry but also adds to the complexity and cost of mastitis treatment. This study was aimed to evaluate how selective stress induced by benzalkonium chloride (BC) promotes antibiotic non-susceptibility in Staphylococcus spp. In addition, we investigated the efficacy of photodynamic inactivation (PDI) in both resistant and susceptible strains. The study determined the minimum inhibitory concentration (MIC) of BC using the broth microdilution method for different Staphylococcus strains. The experiments involved pairing strains carrying the qacA/qacC resistance genes with susceptible strains and exposing them to subinhibitory concentrations of BC for 72 h. The recovered isolates were tested for MIC BC and subjected to disc diffusion tests to assess changes in susceptibility patterns. The results demonstrated that subinhibitory concentrations of BC could select strains with reduced susceptibility and antibiotic resistance, particularly in the presence of S. pasteuri. The results of PDI mediated by toluidine blue (100 µM) followed by 60 min irradiation (total light dose of 2.5 J/cm2) were highly effective, showing complete inactivation for some bacterial strains and a reduction of up to 5 logs in others. [ABSTRACT FROM AUTHOR]

The aims of this study were to investigate the sensitivity and specificity of toluidine blue and/or vinegar in oral cancer screening and to examine the correlation between clinical screening using toluidine blue and vinegar and the expression of the tumor marker p53 and proliferation marker Ki67, respectively. The study consisted of 87 patients with suspected oral squamous cell carcinoma lesions. Toluidine blue and/or vinegar were applied to the lesions, followed by biopsies. The tissues were diagnosed histopathologically and underwent immunohistochemical process for p53 or Ki67. The results revealed that the sensitivity and specificity of oral cancer screening using toluidine blue were 93% and 46%, respectively; whereas the sensitivity and specificity using vinegar were 85% and 81%, respectively. A statistically significant correlation between the use of vinegar and the expression of Ki67 (p = 0.019) was observed. Although there was a difference in the expression of p53 between specimens that were positive and negative to toluidine blue, the correlation did not reach a significant level. Based on the results from this study, vinegar has a lower sensitivity than toluidine blue but a higher specificity for oral cancer screening. The results of the clinical screening using vinegar correlated with the expression of Ki67 at the cellular level.

Telocytes (TCs), a recently discovered special type of stromal cells, have been identified in many organs of many species, including the female and male reproductive system, with proposed multiple potential bio-functions such as homeostasis, immunomodulation, tissue remodeling and regeneration, embryogenesis, angiogenesis and even tumorigenesis. The aim of this study was to investigate the existence, and characteristics of telocytes in normal equine oviduct. To identify them, we used routine light microscopy, non-conventional light microscopy (NCLM), transmission electron microscopy (TEM), and immunohistochemistry. We found that telocytes of the equine oviduct can be recognized in fixed specimens by light microscopy (methylene blue staining), with more details on Epon semi-thin sections (toluidine blue staining) by NCLM, and that they showed positive immunostaining for CD34. The telocytes, with their typical long and moniliform prolongations, formed networks in the stromal space of the submucosa, muscular and serosa layers, particularly in the lamina propia where they were observed in greater quantity. By TEM we have also confirmed the presence of cells ultrastructurally identifiable as telocytes (cells with telopodes alternating podomers and podoms) in the aforementioned locations. Direct intercellular contacts between epithelial cells and neighboring telocytes were evidenced. EIn conclusion, we demonstrated that telocytes are present in the equine oviduct as previously reported in other species. The potential implication of telocytes in multiple potential functions of physiological and pathological processes deserves further investigation. • Cells compatible with telocytes were identified in the equine oviduct. • Cells ultrastructurally identifiable as telocytes were confirmed by TEM. • Telocytes formed networks in the submucosa, especially in the lamina propia. • Intercellular contacts between epithelial cells and telocytes were evidenced. • Role of telocytes in physiological and pathological events needs investigation. [ABSTRACT FROM AUTHOR]

Saponaria sicula Raf. grows in Sicily, Sardinia, and Algeria on limestone cliffs and volcanic sands 1300–2500 m above sea level. The aim of the present study was to investigate how the pedo-climatic conditions influence the micromorphological, phytochemical, and biological properties of Sicilian S. sicula leaves collected in the Madonie Mountains (SsM) and on Etna Mt (SsE). Micromorphological investigations revealed that leaves from SsM had a higher amount of calcium oxalate druses in the mesophyll and a more intense blue–green staining with Toluidine blue O, indicating a higher content of polyphenols. These data were confirmed by phytochemical analyses carried out on hydroalcoholic extracts, which showed a higher content of total phenols (8.56 ± 0.57 g GAE/100 g DE) and flavonoids (6.09 ± 0.17 g RE/100 g DE) in SsM. Sixty-four compounds were identified by LC-DAD-ESI-MS analysis with propelargonidin dimer as the most abundant compound (10.49% and 10.19% in SsM and SsE, respectively). The higher polyphenol content of SsM leaves matches also with their biological activity, identifying SsM extract as the strongest plant complex (IC50 2.75–477.30 µg/mL). In conclusion, the present study experimentally demonstrates that not only climatic differences but also soil characteristics affect the micromorphological, phytochemical, and biological features of this plant species. [ABSTRACT FROM AUTHOR]

The recent findings showed that the superficial fascia is a fibrous layer in the middle of hypodermis, richly innervated and vascularized, and more complex than so far demonstrated. This study showed the presence of mast cells in the superficial fascia of the human abdomen wall of three adult volunteer patients (mean age 42 ± 4 years; 2 females, 1 male), by Toluidine Blue and Safranin-O stains and Transmission Electron Microscopy. Mast cells are distributed among the collagen bundles and the elastic fibers, near the vessels and close to the nerves supplying the tissue, with an average density of 20.4 ± 9.4/mm2. The demonstration of the presence of mast cells in the human superficial fascia highlights the possible involvement of the tissue in the inflammatory process, and in tissue healing and regeneration processes. A clear knowledge of this anatomical structure of the hypodermis is fundamental for a good comprehension of some fascial dysfunctions and for a better-targeted clinical practice. [ABSTRACT FROM AUTHOR]

Background: To evaluate the quantity and quality of bone in the newly formed edentulous area produced by the orthodontic implant site-switching technique. Methods: The bilateral maxillary first premolars of five beagle dogs were extracted and bone defects were created. The right and left sides of the maxilla were randomly divided into control and experimental sides. On the experimental side, the maxillary second premolar was mesially moved into the position of the missing first premolar. On the control side, the second maxillary premolar was extracted. Six months later, the beagles were euthanized. Microcomputer tomography was used to analyze bone microstructure parameters, alveolar bone height and alveolar bone width of the regenerated bone. Histological analysis was performed by staining tissue sections with toluidine blue. Results: Median BV/TV values in the experimental group (81.78%) were significantly larger than those in the control group (35.67%; p = 0.04). Median Tb.Sp values in the experimental group (0.14 mm) were significantly lower than those in the control group (0.54 mm; p = 0.04). Median Tb.Th values in the experimental group (0.48 mm) were significantly higher than those in the control group (0.21 mm; p = 0.04). Median Tb.Pf values in the experimental group (0.65/mm) were significantly lower than those in the control group (3.15/mm; p = 0.04). There was no significant difference in the trabecular number (Tb.N) between the two groups (p = 0.23). The median alveolar bone height values in the experimental group (-0.81 mm) were significantly higher than those in the control group (-2.11 mm; p = 0.04) at a distance 5 mm from the mesial CEJ of the third premolar. The median alveolar bone height values in the experimental group (0.45 mm) were significantly higher than those in the control group (-1.70 mm; p = 0.04) at a distance 6 mm from the mesial CEJ of the third premolar. There was no significant difference in alveolar bone width when compared between the two groups (p > 0.05). Conclusions: The newly formed edentulous area created by orthodontic treatment had more compact and thicker trabeculae than the extraction socket. Furthermore, the newly formed edentulous area had a greater alveolar bone height available for the placement of implants. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the influence of IR (λ850 ± 10 nm) and violet (λ405 ± 10 nm) LED phototherapy on total mast cells counts and its ability to influence mast cell degranulation. For this, 27 Wistar rats were used and were randomly distributed into three groups: control, IR LED, and violet LED. When indicated, irradiation done and they were sacrificed, had their tongue removed immediately, 20-min, 45-min, and 2-h after irradiation. Samples were processed to wax, cut, and stained with Toluidine Blue. Intact and degranulated mast cells were counted under light microscopy, and statistical analysis was carried out. In the superficial connective tissue and muscular tissues, violet LED light caused a significant increase in both total number and degranulated mast cells when compared to the control group immediately after irradiation. The degranulation indexes were higher in the groups irradiated with Violet light, both in superficial connective tissue and muscular tissues in relation to the timing. Irradiation with IR LED caused immediate increase in the total number and degranulated of mast cells when compared to the control group only in the superficial connective tissue. In all times observed, the highest total amount of mast cells was seen immediately after irradiation, except in the muscular tissue, which presented the highest amount after 20-min. It was concluded that IR and violet LED light were able to increase the number of mast cells and inducing degranulation in oral mucosa. However, considering that violet LED light can be harmful in periodontal disease, it seems that the use of IR LED light could be the best option in Dentistry. [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]

Mammalian pineal gland, an integral part of brain, is a photo-neuroendocrine gland which controls the circadian rhythms and reproductive activity in seasonal breeders. This study was designed to evaluate the gross morphological and histological changes in pineal gland and plasma melatonin profile of Beetal goat in relation to different age groups. For this purpose, a total of 24 healthy female Beetal goats were selected from the local abattoir of Faisalabad and were divided into three age groups naming, prepubertal, pubertal and postpubertal (n=8). Before slaughtering, 10 ml blood was collected in EDTA coated tubes for determination of plasma melatonin level using ELISA kit. After slaughtering, heads of the animals were dissected, pineal gland was enucleated and gross morphometric measurements of the gland were taken. For light microscopic studies, the pineal tissues were fixed in a solution of 10% neutral buffered formalin and processed by the routine tissue paraffin sectioning technique. The tissue sections were stained with Hematoxylin and Eosin, modified von Kossa stain and toluidine blue. The results revealed that age of the animals has a significant effect (P≤0.05) on gross anatomical parameters of pineal gland (weight, length, width and volume) as well as histomorphometric parameters including number and volume of pinealocytes, relative area of parenchyma, brain sand, connective tissue and blood vessels of the pineal gland. Age also affected the plasma melatonin level and was found significantly highest (5.63±0.20 pg/ml) in the pubertal group. In conclusion, the present study revealed that age influences the gross and histomorphology along with plasma melatonin level of the Beetal goat. [ABSTRACT FROM AUTHOR]

Stem cells from the apical papilla (SCAPs) are used to regulate the microenvironment of nerve defects. KDM6B, which functions as an H3K27me3 demethylase, is known to play a crucial role in neurogenesis. However, the mechanism by which KDM6B influences the neurogenesis potential of SCAPs remains unclear. We evaluated the expression of neural markers in SCAPs by using real-time RT-PCR and immunofluorescence staining. To assess the effectiveness of SCAP transplantation in the SCI model, we used the BBB scale to evaluate motor function. Additionally, toluidine blue staining and Immunofluorescence staining of NCAM, NEFM, β-III-tubulin, and Nestin were used to assess nerve tissue remodeling. Further analysis was conducted through Microarray analysis and ChIP assay to study the molecular mechanisms. Our results show that KDM6B inhibits the expression of NeuroD, TH, β-III tubulin, and Nestin. In vivo studies indicate that the SCAP-KDM6Bsh group is highly effective in restoring spinal cord structure and motor function in rats suffering from SCI. Our findings suggest that KDM6B directly binds to the HES1 promoter via regulating H3K27me3 and HES1 expression. In conclusion, our study can help understand the regulatory role of KDM6B in neurogenesis and provide more effective treatments for nerve injury. [ABSTRACT FROM AUTHOR]

Background: Diagnostic adjuncts such as toluidine blue have been investigated as screening tools that improve visual examination of potentially malignant disorders (PMD) and oral cancer. Acetic acid has been reported to be of value in the early detection of cervical cancers. This study assessed the utility of 5% acetic acid as a diagnostic adjunct in oral PMD and compared the accuracy of acetic acid with toluidine blue in the detection of dysplastic PMD and high-risk lesions. Materials and Methods: This cross-sectional study was conducted at a dental hospital in a rural setting. Thirty-one patients with oral PMD formed the study group. Five percent acetic acid was applied to the lesions, followed by toluidine blue application and biopsy. Sensitivity, specificity, and positive and negative predictive values were computed considering true positives as stain uptake in dysplastic and high-risk PMD. Results: The sensitivity, specificity, and positive and negative predictive values of acetic acid for identifying dysplastic or malignant lesions were 100%, 13.3%, 51.2%, and 100%, respectively, and that for toluidine blue were 75%, 100%, 100%, and 78.9%, respectively. The corresponding values for identifying high-risk PMD (lesions with moderate and severe dysplasia) using acetic acid were 100%, 9.1%, 25.9%, and 100%, respectively, and for toluidine blue were 85.7%, 81.8%, 60%, and 94.7%, respectively. Conclusion: The utility of acetic acid in detecting dysplasia and high-risk PMD is severely limited due to its poor specificity. Compared with acetic acid, toluidine blue is a superior screening tool. [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]

PURPOSE OF THE STUDY Articular cartilage injury is a common disease in daily life, with a high incidence. The aim of this study was to investigate the effect and mechanism of miRNA-140-3p in bone mesenchymal stem cells (BMSCs)-derived exosomes under hypoxia on inflammatory articular chondrocytes. MATERIAL AND METHODS To simulate the pathological status of arthritis, rat chondrocytes were used to establish the osteoarthritis (OA) model by IL-1β (10 μg/ml) as a modulating in vitro, and exosomes were isolated by differential ultra-high speed centrifugation. The cell counting kit-8, wound healing and flow cytometry assays were utilized to assess proliferation, migration and apoptosis of chondrocytes, respectively. Lipogenic and chondrogenic differentiation of chondrocytes were detected by oil red O staining and toluidine blue staining individually. The expressions of miR-140-3p and chondrocyte-specific gene mRNA were investigated using qRT–PCR. Western blot was applied to assess chondrocyte associated proteins and BMSC-Exo surface protein markers, and immunohistochemistry was adopted to detect the staining of collagen I and II. RESULTS Under scanning electronic microscope, the shape of exosomes was almost round. Exosome treatment prominently impaired the inhibition of chondrocytes’ proliferative and migrative ability by IL-1β. It was found hypoxia had a more marked impact on proliferation, expression of collagen II and apoptosis in OA chondrocytes than normoxia, as well as a stronger effect on weakening adipose differentiation and enhancing chondrogenic differentiation in inflammatory chondrocytes. Furthermore, incubation with BMSC-Exo overexpressing miR-140-3p can remarkably increase the survival rate and migration in inflammatory chondrocytes. In addition, overexpression of miR-140-3p was found to enhance the chondrogenic differentiation of inflammatory chondrocytes. Furthermore, we found that the healing effect of exosomes on inflammatory chondrocytes under hypoxic conditions was produced by a rise in miR-140-3p expression within them and that hypoxia-mediated upregulation of miR-140-3p expression occurred through HIF-1α. CONCLUSIONS Under hypoxia, BMSC-Exo enhanced the chondrogenic phenotype, increased the viability of inflammatory chondrocytes. The overexpression of miR-140-3p in BMSC-Exo is beneficial to protect joints and delaying the pathogenesis in OA. [ABSTRACT FROM AUTHOR]

In this paper, a new multifunctional nanocomposite was synthesized to combine both the advantages of sorption properties and antibacterial efficiency. The thiourea polyurethane/zinc oxide nanocomposite was prepared by coupling thiourea polyurethane foam and zinc oxide nanoparticles. The thiourea polyurethane/zinc oxide was characterized using several tools. The change in the surface chemistry of the thiourea polyurethane/zinc oxide nanocomposite (surface area was increased from 2.8 to 10.6 m2/g), due to the attached functional groups, increased its sorption properties. Also, the thiourea polyurethane/zinc oxide showed a crystalline phase, embedded inside the amorphous structure, with good thermal stability to 230 °C. The pore volume and radius of thiourea polyurethane/zinc oxide were 0.017 cm3/g and 3.4 nm with mesopores/micropores ratio of 11:1. It combines collecting pollutants on its surface, the ability to degrade these pollutants, and a good antibacterial effect. Thiourea polyurethane/zinc oxide was applied for the removing toluidine blue and brilliant green basic dyes from the wastewater. Several parameters such as pH, dye concentration, temperature, and time were studied. The removal percentages of brilliant green and toluidine blue dyes from the wastewater were 92.4–98.2% at pH 3–12. Also, the removing rate of the dyes was very fast (3–5 min) at 25 °C. The estimated capacities of new nanocomposite for toluidine blue and brilliant green were 92 and 227 mg/g. The removal process was spontaneous (ΔG was − 7.3 and − 8.6). As a result, the applicability of thiourea polyurethane/zinc oxide as an effective renewable antibacterial sorbent for removing cationic dyes was maintained. [ABSTRACT FROM AUTHOR]

Background: Despite intensive research, the ideal protocol applied to maximize the overall benefits of antimicrobial photodynamic therapy (aPDT) remains unexplored. Evidence exists that following aPDT, the diffused light beyond the photosensitizer can exert a secondary therapeutic effect known as photobiomodulation (PBM), which stimulates the healing of the surrounding tissues. Therefore, the aim of this study was to examine the attenuation properties of five different photosensitizers activated by their corresponding laser wavelengths. Methods: The illumination of various concentrations of chosen photosensitizers, curcumin, methylene blue, toluidine blue, indocyanine green and a methylene blue derivative, irradiated by their respective laser wavelengths (445 nm, 635 nm, 660 nm and 808 nm) was explored via a spectrophotometric analysis. The onward transmitted light intensities for each combination of a photosensitizer and laser wavelength were assessed. The attenuation percentages observed were statistically evaluated using an analysis-of-variance (ANOVA) model. A Tukey's post hoc test was performed to determine the significance of differences between individual group mean values. Results: With the exception of toluidine blue illuminated by an 808 nm laser, which showed the lowest intensity loss, all the other photosensitizers presented an attenuation range of 63% to 99%. Conclusions: At appropriate concentrations, all the examined photosensitizers may allow the passage of sufficient wavelength-dependent light transmission. Calculated fluences are proposed to achieve secondary, beneficial PBM effects. [ABSTRACT FROM AUTHOR]

Objectives: Odontogenic lesions include a broad range of pathologies that depend upon an inductive process toward odontogenesis before hamartomatous/neoplastic growth. The epithelium and stroma have an imperative role in pathogenesis and biological behavior. Hence, the study aims to evaluate the biology of the epithelium and stroma in odontogenic cysts (OCs). Material and Methods: Histochemical analysis was done (Hematoxylin and Eosin stain, Papanicolaou stain, toluidine blue, and picrosirius red) to determine the presence of keratin, epithelial thickness, mast cells (MCs), and the nature of collagen in OCs. Results: Keratin was absent in highly inflamed cases. MCs play a crucial role in cyst pathogenesis. In addition, polarizing microscopy helps in depicting the pattern of collagen fibers. Conclusion: The epithelium and the nature of stroma have an imperative role in the biological behavior of OCs. [ABSTRACT FROM AUTHOR]

Purpose: This study aims to investigate the numerical increase, localization, granulation status, and immunophenotypic properties of mast cells (MCs) in epineurectomy-induced nerve damage and lipopolysaccharide (LPS)-induced systemic infection models. Materials and Methods: In this study, the animals were divided into three groups of 6 each. One of the groups was determined as the control group, epineurectomy was applied to one group, and systemic inflammation was created by regular LPS injections in the other group. Then, the obtained nerve tissues were stained histochemically with Hematoxylin and Eosin toluidine blue, and the increase, localization, and granulation status of MCs were examined. Immunohistochemically, antitryptase and antichymase staining were performed to determine the immunophenotypes of MCs. Results: As a result, while the number of MCs increased in both groups compared to the control group, MCs in the LPS group were in the epineurium, and MCs in the epineurotomy group were located between the nerve fibers. While MCs in the LPS group showed very severe degranulation, mild degranulation was observed in the epineurotomy group, and almost no degranulated MCs were observed in the control group. Conclusion: This study is critical because it is one of the first studies to compare MCs in different nerve damage types and examine the expression of chymase and tryptase.

Human epididymis protein 4 (HE4) is a highly sensitive and specific biomarker for diagnosing ovarian cancer. In this work, a new electrochemical aptasensor for rapid and sensitive detection of HE4 was constructed. Firstly, metal–organic framework/ketjen black (NH2-MIL-53(Al)/KB) composite with abundant amino groups and large specific surface area was prepared as signal amplifier to anchor gold nanoparticles (AuNPs). Subsequently, signal probe (SP) and a large amount of electroactive toluidine blue (Tb) were attached on AuNPs to form the tracer label, producing a distinct electrochemical detection signal. Moreover, Ni3(HITP)2 was modified by poly(diallyldimethylammonium chloride, PDDA) to obtain PDDA/Ni3(HITP)2, and then it was covered with deposited gold nanocrystals (DpAu) to form the sensing platform, which increased the electrical conductivity, specific surface area and the loading of triple-helix assembled probes (TAP). Target HE4 was bound to its aptamer in the outer layer of the TAP, leaving a separate capture probe (CP) on the electrode surface that can bind with the tracer label. The proposed aptasensor for HE4 detection showed a wide linear range from 1 to 10 nM with a detection limit of 0.41 fM. In addition, the aptasensor has good selectivity, stability and reproducibility. More importantly, this method showed satisfactory results in clinical serum sample analysis compared with ELISA, possessing application prospects in ovarian cancer diagnosis. [ABSTRACT FROM AUTHOR]

Mast cells and the mediators they release play a major role in the pathogenesis of lung diseases. They regulate the function of the smooth muscle layer of the pulmonary airways and the same layer of the blood vessels. There are no data about the distribution of metachromatic and tryptase-positive mast cells in the blood vessels' wall as well as on the content of ghrelin in mast cell granules in rat's lung. The aim of this research was to present an analysis of the number and localisation of tryptasepositive, ghrelin-positive and metachromatic mast cells in the blood vessels (arteries, capillaries, venules and veins) in rats of different age. Six male Wistar rats were used for each age group? 20 days, 3 months and 1 year. Tissue slices for histochemical examination were taken from the caudal lobe of the left lung and stained with toluidine blue. Immunohistochemical reactions were then performed to indicate the expression of tryptase and ghrelin, allowing comparing the presence of tryptase-, ghrelin-positive and metachromatic mast cells in the wall of the blood vessels of different diameters. The light microscopical study showed that in all types of blood vessels from the three age groups, the number of ghrelin positive cells was the largest, followed by tryptase-positive and metachromatic mast cells. The observed differences in the distribution of these cells are important for maintaining the lung homeostasis and can be used as reference values in experimental studies in order to obtain accurate results. [ABSTRACT FROM AUTHOR]

Detailed mast cell classification is reported in humans and rats, however such classification is not available in porcine common hepatic duct. It is interesting to find out whether mast cells in common hepatic duct are able to produce ghrelin, which is involved in a series of biological functions including regulation of food intake, body weight, gastrointestinal motility, hormone secretion, glucose release, cardiovascular functions, enzyme release, cell proliferation and reproduction in pigs. Because the determination of the cellular mechanisms responsible for normal and pathological biliary tract motility is difficult in humans, swine appears to be one of the most suitable animal model for physiological and morphological studies related to ghrelin peptide. The lack of information about the distribution of mast cells in the wall of common hepatic duct (Ductus hepaticus communis) in domestic swine motivated us to undertake this study in order to gain a better understanding of ghrelin peptide. The aim of this study was to define the localisation and number of tryptase-, ghrelin- and toluidine blue positive mast cells in the layers of porcine common hepatic duct. In this respect, the immunohistochemical staining for detection of tryptase and ghrelin expression was performed. The toluidine blue staining allowed the visualisation of metachromatic mast cells. The comparative study showed that tryptase positive cells were more numerous than metachromatic cells in all layers of the studied organ but the density of ghrelin positive cells was the highest. Tryptase immunohistochemistry allowed distinguishing ghrelin expressing mast cells from all ghrelin positive cells. The highest number of ghrelin positive cells indicated that not only mast cells but other cells also express ghrelin. The results allowed concluding that studied mast cell types had different density in the three main layers of common hepatic duct. The highest density of mast cells in the muscular layer and the possibility of mast cells to express ghrelin define its role in regulation of common hepatic duct motility and glandular secretion. [ABSTRACT FROM AUTHOR]

In this study, we aimed to verify the efficacy of the newly developed β-tricalcium phosphate (β-TCP) block as a substitute for autologous bone blocks using a dog model. A total of 13 male beagle dogs (weighing 9.83±0.49 kg) 12 to 18 months old were used. A standardized bone defect 15 mm wide, 10 mm high, and 3 mm deep was created in dogs 2 months after extraction of the mandibular fourth premolar and first posterior molar. To determine its suitability as a bone substitute, the β-TCP block was transplanted into the dental socket. The control groups underwent only bone defect creation. 8 and 16 weeks after transplantation, an impression of the mandibular bone was taken using alginate, and plaster models were made. To analyze alveolar bone volume, the plaster models were scanned using a 3D scanner. For histopathological examination, mandibular bones were embedded in resin, and sections were stained with toluidine blue. At 8 and 16 weeks, the width of the alveolar crest in the control group was reduced compared to the preoperative width at all measurement points (p<0.05). Osteoblasts and osteoclasts were observed around the transplanted area in the experimental group. The area fraction of new bone increased from 9.35% to 29.72% and the residual β-TCP area fraction decreased from 23.63% to 13.14% from 8 to 16 weeks after surgery, with significant differences observed. In conclusion, β-TCP can be formed into a block shape with high excisibility. β-TCP has both compressive strength and good resorptive and replacement properties while preserving 60% porosity, allowing it to maintain mechanical strength for an extended duration. [ABSTRACT FROM AUTHOR]

Copyright of Acta Materiae Compositae Sinica is the property of Acta Materiea Compositae Sinica Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background. Diabetic neuropathic osteoarthropathy (DNOAP) is a rare and easily missed complication for diabetes that leads to increased morbidity and mortality. DNOAP is characterized by progressive destruction of bone and joint, but its pathogenesis remains elusive. We herein aimed to investigate the pathological features and pathogenesis of the cartilages damage in DNOAP patients. Methods. The articular cartilages of eight patients with DNOAP and eight normal controls were included. Masson staining and safranine O/fixed green staining (S-O) were used to observe the histopathological characteristics of cartilage. The ultrastructure and morphology of chondrocytes were detected by electron microscopy and toluidine blue staining. Chondrocytes were isolated from DNOAP group and control group. The expression of receptor activator of nuclear factor kappaB ligand (RANKL), osteoprotegerin (OPG), interleukin-1 beta (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and Aggrecan protein was evaluated by western blot. Reactive oxygen species (ROS) levels were measured using a 2′,7′-dichlorofluorescin diacetate (DCFH-DA) probe. The percentage of apoptotic cells was determined by flow cytometry (FCM). The chondrocytes were cultured with different glucose concentrations to observe the expression of RANKL and OPG. Results. Compared with the control group, the DNOAP group showed fewer chondrocytes, subchondral bone hyperplasia, and structural disorder, and a large number of osteoclasts formed in the subchondral bone area. Moreover, mitochondrial and endoplasmic reticulum swellings were observed in the DNOAP chondrocytes. The chromatin was partially broken and concentrated at the edge of nuclear membrane. The ROS fluorescence intensity of chondrocyte in DNOAP group was higher than that in normal control group (28.1 ± 2.3 vs. 11.9 ± 0.7 ; P < 0.05). The expression of RANKL, TNF-α, IL-1β, and IL-6 protein in DNOAP group was higher than that in normal control group, whereas OPG and Aggrecan protein were lower than that in normal control group (both P < 0.05). FCM showed that the apoptotic rate of chondrocyte in DNOAP group was higher than that in normal control group (P < 0.05). The RANKL/OPG ratio showed significant upward trend when the concentration of glucose was over than 15 mM. Conclusions. DNOAP patients tend to have severe destruction of articular cartilage and collapse of organelle structure including mitochondrion and endoplasm reticulum. Indicators of bone metabolism (RANKL and OPG) and inflammatory cytokines (IL-1β, IL-6, and TNF-α) play an important role in promoting the pathogenesis of DNOAP. The glucose concentration higher than 15 mM made the RANKL/OPG ratio change rapidly. [ABSTRACT FROM AUTHOR]

Although the fabrication of controllable three-dimensional (3D) microstructures on substrates has been proposed as an effective solution for SERS, there remains a gap in the detection and manufacturability of 3D substrates with high performance. In this study, photolithography is adopted to obtain a pyramid-like array on a patterned sapphire substrate (PSS), with Al2O3 as the dielectric layer. In addition, silver nanoparticles (AgNPs) are used to decorate Au films to obtain mass-producible 3D SRES substrates. In the case of low fluorescence, the substrate realizes the coupling of localized surface plasmon polaritons (LSPs) and surface plasmon polaritons (SPPs), which is consistent with the simulation results obtained using the finite element method. The performance of the SERS substrate is evaluated using rhodamine 6G (R6G) and toluidine blue (TB) as probe molecules with detection limits of 10−11 M and 10−9 M, respectively. The substrate exhibits high hydrophobicity and excellent light-capturing capability. Moreover, it shows self-cleaning ability and long-term stability in practical applications. Allowing for the consistency of the composite substrate in the preparation process and the high reproducibility of the test results, it is considered to be promising for mass production. [ABSTRACT FROM AUTHOR]

INTRODUCTION: Mast cells present in the connective tissue stroma release pro inflammatory and mitogenic cytokines. The mast cell in tumor act as a host immunologic anti tumor response. More than 90% of oral cancers are squamous cell carcinomas with oral epithelial dysplasia being the most common potentially malignant disorder. Mast cells play an important role in tumourigenesis. In some studies, with different granules of mast cell is closely related with angiogenesis and tumor invasion. In this study we used toluidine blue for the staining because it reveals mast cells as large, purple, oval and granulated cells. MATERIAL AND METHODS: 30 cases of oral epithelial dysplasia, 30 cases of Oral squamous cell carcinoma (OSCC) and 10 cases of Normal oral mucosa (NOM) were studied for mast cell number using toluidine blue. Mast cells were counted using an Olympus CX41 microscope fitted with an Olympus oculometer grid. Counting was carried out at ×40 by two independent observers and was done in six non overlapping fields in each slide. Mast cells were identified on the basis of the purple color attained by the granules after toluidine blue staining, but the nucleus of these cells appeared blue. RESULTS: Highly significant increase of mast cells in oral epithelial dysplasia on comparison with OSCC whereas there was only a significant increase in mast cells in OSCC on comparison with NOM. CONCLUSION: Mast cells can be used an indicator of increased angiogenesis and can help in the prediction of carcinogenesis, its progression, and also in the prognosis of the malignant lesions. [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]

The main target of bone tissue engineering is to design biomaterials that support bone regeneration and vascularization. Nanostructured membranes of (MMA)1-co-(HEMA)1/(MA)3-co-(HEA)2 loaded with 5% wt of SiO2-nanoparticles (Si-M) were doped with zinc (Zn-Si-M) or doxycycline (Dox-Si-M). Critical bone defects were effectuated on six New Zealand-bred rabbit skulls and then they were covered with the membranes. After six weeks, a histological analysis (toluidine blue technique) was employed to determine bone cell population as osteoblasts, osteoclasts, osteocytes, M1 and M2 macrophages and vasculature. Membranes covering the bone defect determined a higher count of bone cells and blood vessels than in the sham group at the top regions of the defect. Pro-inflammatory M1 appeared in a higher number in the top regions than in the bottom regions, when Si-M and Dox-Si-M were used. Samples treated with Dox-Si-M showed a higher amount of anti-inflammatory and pro-regenerative M2 macrophages. The M1/M2 ratio obtained its lowest value in the absence of membranes. On the top regions, osteoblasts were more abundant when using Si-M and Zn-Si-M. Osteoclasts were equally distributed at the central and lateral regions. The sham group and samples treated with Zn-Si-M attained a higher number of osteocytes at the top regions. A preferential osteoconductive, osteoinductive and angiogenic clinical environment was created in the vicinity of the membrane placed on critical bone defects. [ABSTRACT FROM AUTHOR]

Copyright of Wool Textile Journal is the property of National Wool Textile Science & Technology Information Center and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective: In order to establish a non-decalcified pathological section preparation method for bone tissue specimens of lumbar vertebral body of miniature pigs implanted with bone repair materials. Methods: In this study, lumbar vertebral bone tissue specimens containing bone repair materials were segmented to expose tissue sections. Dehydration with graded ethanol followed by infiltration with Technovit 7200 VLC photopolymer resin. Photopolymerization embedding of lumbar vertebral body tissue specimens containing bone repair material by co-irradiation with yellow and blue light, Preparation of non-decalcified pathological tissue sections with bone-containing repair materials by means of hard tissue pathological cutting and grinding system. Results: The results show that the pathological tissue sections prepared by the above method, After HE staining and toluidine blue staining, observation under a light microscope can better show various tissue and cell structures of bone, The direction and connection of trabecular bone can be clearly observed. Conclusions: A method for preparing pathological tissue sections of bone tissue specimens containing bone repair materials was established, Realized the preparation of histopathological sections of non-decalcified bone with bone repair material, Histological observation with implants achieved after pathological staining, It provides a new pathological detection method and histological evaluation method for the animal test research of biological materials and medical devices. [ABSTRACT FROM AUTHOR]

A semi-interpenetrating network consisting of karaya gum-graft-polyacrylamide and poly(vinyl alcohol) has been developed as an adsorbent material for dye removal applications. The system was made by microwave irradiation of the aqueous mixture of karaya gum, acrylamide, N,N′-methylenebisacrylamide, and poly (vinyl alcohol); and was characterized by FTIR, TGA, SEM, and XRD techniques. It was evaluated as an adsorbent material for removal of ionic dyes from aqueous solutions. Appreciably high adsorption capacity is exhibited by the material towards cationic dyes as indicated by values of maximum adsorption capacity of 82.28, 72.94, 48.75 and 34.67 mg/g towards adsorption of cationic dyes, namely methylene blue (MB), crystal violet (CV), rhodamine B (Rh B), and toluidine blue (TB). Relatively low value of 21.66 mg/g is obtained for anionic dye indigo carmine (IC). The kinetic data of dye adsorption best fitted with the pseudo-first-order kinetic model with R2 values of 0.979, 0.989, 0.946, 0.991, and 0.986 for TB, MB, CV, Rh B and IC, respectively. The isotherm data fit well with the Freundlich model with R2 values of 0.986, 0.955, 0.990, 0.980 and 0.993 for TB, MB, CV, Rh B, and IC, respectively, indicating the heterogeneous and multilayer adsorption. The thermodynamic studies showed dye adsorption to be an exothermic and spontaneous process. [ABSTRACT FROM AUTHOR]

Background/Aims: Alzheimer's disease is a progressive neurological disorder characterized by the intracellular accumulation of Tau protein aggregates. In the present work, we studied the effect of Toluidine Blue and photo-excited Toluidine Blue on the aggregation of repeat Tau using in vitro assays. Methods: The in vitro experiments were carried out on recombinant repeat Tau which was purified by cation exchange chromatography. The ThS fluorescence analysis was used to study the aggregation kinetics of Tau. CD spectroscopy and electron microscopy were used to study the secondary structure and morphology of Tau respectively. The actin cytoskeleton modulation was studied in Neuro2a cells with help of immunofluorescent microscopy. Results: Results showed that Toluidine Blue efficiently inhibited the formation of higher-order aggregates, which was evidenced by Thioflavin S fluorescence assay, SDSPAGE, and TEM. Immunofluorescence studies on the cytoskeleton of Neuro2a cells showed that Toluidine Blue and photo-excited Toluidine Blue treatment at a non-toxic concentration of 0.5 µM stimulated the formation of actin-rich lamellipodia and filopodia structures. Tubulin networks were also differentially modulated after the treatment of Toluidine Blue and photoexcited Toluidine Blue. End-binding protein 1 (EB1) levels were observed to increase after Toluidine Blue and photo-excited Toluidine Blue treatment indicating accelerated microtubule polymerization. Conclusion: The overall study suggested that Toluidine Blue inhibited the aggregation of soluble Tau and photo-excited Toluidine Blue disaggregated the pre-formed Tau filaments. In our study, TB and PE-TB were observed to be potent against Tau aggregation. We observed a distinctive modulation of actin, tubulin networks, and EB1 levels after TB and PE-TB treatment, which suggested that TB and PE-TB have potency against cytoskeleton deformities. [ABSTRACT FROM AUTHOR]

Introduction: Chronic diseases have become major health issues encountering people globally and on top of the list comes "Diabetes Mellitus (DM)". One of its serious complications is "Diabetic Nephropathy (DN)". Aim of the Work: Demonstration of the possible effect of pentoxifylline (PTX) on induced diabetic nephropathy in rats as a cure or prophylactic therapy. Materials and Methods: Seventy adult male albino rats were randomly distributed into five groups; group I (control group), group II (PTX group), group III (diabetic nephropathy group), group IV (diabetic rats received pentoxifylline as a prophylactic treatment one week after induction of diabetes) and group V (diabetic rats received pentoxifylline as a curative treatment starting after sixth week of induction of diabetes). Induction of diabetes was done by intraperitoneal injection of a single dose of streptozotocin (50 mg/kg). A single dose of PTX (200 mg/kg) was given daily by gastric intubation. At the end of experiment (after 8 weeks), renal specimens were collected and processed for paraffin blocks and stained with H&E and immunohistochemical stains and examined by light microscope. Other samples were prepared to obtain semithin sections stained with toluidine blue to be examined by light microscope and ultrathin sections to be examined by transmission electron microscope (TEM). Results: In diabetic nephropathy, there were shrunken glomeruli. The epithelial cells of renal tubules showed vacuolations, degenerative changes and complete shedding. There was anti-NF-κb strong positive reaction of renal tubules. In TEM examination, there were thickening of the glomerular filtration barrier, fusion of foot processes of podocytes, damage of renal tubules. Histopathological examination of the kidney showed improvement in the prophylactic and the curative groups. There was a significant decrease in anti-NF-κb positive reaction in groups II, IV and V. Conclusion: Pentoxifylline had both prophylactic and curative effects against diabetic nephropathy. [ABSTRACT FROM AUTHOR]

Due to the similar pattern of ghrelin localisation in human and porcine intestine, swine are described as a suitable animal model for experiments related to ghrelin and its receptors. In a previous study of ours, the presence of ghrelin immunoreactive endocrine cells in common bile duct (Ductus choledochus) (CBD) of domestic swine was established. However, data regarding the distribution of mast cell types in the CBD in this species are missing. The current study aimed to determine the distribution of tryptase-, ghrelin- and toluidine blue positive mast cells in the wall of extra- and intramural parts of porcine CBD. For this purpose, immunohistochemical staining for detection of tryptase was performed in order to identify ghrelin reactivity in mast cells. Additionally, toluidine blue staining was carried out to compare the number of metachromatic mast cells with that of tryptase positive cells. The comparative quantitative analysis showed that the number of tryptase immunorective cells was higher than that of metachromatic ones in the CBD wall. Ghrelin immunoreactive cells were the most numerous, therefore it can be assumed that not only mast cells but other cells also contain ghrelin. The three mast cells types were more abundant in the propria and subserosa of the intramural part than in the same layers of extramural CBD part. In the muscle layer, mast cell number was higher in the extramural than in the intramural part of the duct. In conclusion, taking into account the results obtained, an important role of mast cells play in controlling CBD motility and biliary gland function was suggested. [ABSTRACT FROM AUTHOR]

• Synovitis was more severe in knees subjected to synovium harvesting than in knees not subjected to harvesting. • Healing was better in the menisci that underwent mesenchymal stem cell (MSC) transplantation than in the control menisci that did not undergo MSC transplantation. • Menisci treated with autologous MSCs showed no red granulation at the tear of the meniscus, but menisci not treated with MSCS showed red granulation. • Macroscopic scores, inflammatory cell infiltration scores, and matrix scores assessed by toluidine blue staining were all significantly better in the autologous MSC group than in the control group without MSCs. • Autologous synovial MSC transplantation suppressed the inflammation caused by synovial harvesting and promoted healing of the repaired menisci in micro minipigs. Allogeneic synovial mesenchymal stem cells (MSCs) effectively promote meniscus healing in micro minipigs. We investigated the effect of autologous synovial MSC transplantation on meniscus healing in a micro minipig model of meniscus repair showing synovitis after synovial harvesting. Synovium was harvested from the left knee of the micro minipigs after arthrotomy and used to prepare synovial MSCs. The left medial meniscus in the avascular region was injured, repaired, and transplanted with synovial MSCs. First, synovitis was compared after 6 weeks in knees with and without synovial harvesting. Second, the repaired meniscus was compared for the autologous MSC group and the control group (in which synovium was harvested but MSCs were not transplanted) 4 weeks after transplantation. Synovitis was more severe in knees subjected to synovium harvesting than in knees not subjected to harvesting. Menisci treated with autologous MSCs showed no red granulation at the tear of the meniscus, but menisci not treated with MSCS showed red granulation. Macroscopic scores, inflammatory cell infiltration scores, and matrix scores assessed by toluidine blue staining were all significantly better in the autologous MSC group than in the control group without MSCs (n = 6). Autologous synovial MSC transplantation suppressed the inflammation caused by synovial harvesting in micro minipigs and promoted healing of the repaired meniscus. [Display omitted] [ABSTRACT FROM AUTHOR]

Copyright of Journal of Guangxi Normal University - Natural Science Edition is the property of Gai Kan Bian Wei Hui and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Collagenous tissues exhibit anisotropic optical properties such as birefringence and linear dichroism (LD) as a result of their structurally oriented supraorganization from the nanometer level to the collagen bundle scale. Changes in macromolecular order and in aggregational states can be evaluated in tendon collagen bundles using polarization microscopy. Because there are no reports on the status of the macromolecular organization in tendon explants, the objective of this work was to evaluate the birefringence and LD characteristics of collagen bundles in rat calcaneal tendons cultivated in vitro on substrates that differ in their mechanical stiffness (plastic vs. glass) while accompanying the expected occurrence of cell migration from these structures. Tendon explants from adult male Wistar rats were cultivated for 8 and 12 days on borosilicate glass coverslips (n = 3) and on nonpyrogenic polystyrene plastic dishes (n = 4) and were compared with tendons not cultivated in vitro (n = 3). Birefringence was investigated in unstained tendon sections using high-performance polarization microscopy and image analysis. LD was studied under polarized light in tendon sections stained with the dichroic dyes Ponceau SS and toluidine blue at pH 4.0 to evaluate the orientation of proteins and acid glycosaminoglycans (GAG) macromolecules, respectively. Structural remodeling characterized by the reduction in the macromolecular orientation, aggregation and alignment of collagen bundles, based on decreased average gray values concerned with birefringence intensity, LD and morphological changes, was detected especially in the tendon explants cultivated on the plastic substrate. These changes may have facilitated cell migration from the lateral regions of the explants to the substrates, an event that was observed earlier and more intensely upon tissue cultivation on the plastic substrate. The axial alignment of the migrating cells relative to the explant, which occurred with increased cultivation times, may be due to the mechanosensitive nature of the tenocytes. Collagen fibers possibly played a role as a signal source to cells, a hypothesis that requires further investigation, including studies on the dynamics of cell membrane receptors and cytoskeletal organization, and collagen shearing electrical properties. [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]

Toluidine blue O (TBO) is a phenothiazine dye that, due to its photochemical characteristics and high affinity for biomembranes, has been revealed as a new photosensitizer (PS) option for antimicrobial photodynamic therapy (PDT). This points to a possible association with membranous organelles like mitochondrion. Therefore, here we investigated its effects on mitochondrial bioenergetic functions both in the dark and under photostimulation. Two experimental systems were utilized: (a) isolated rat liver mitochondria and (b) isolated perfused rat liver. Our data revealed that, independently of photostimulation, TBO presented affinity for mitochondria. Under photostimulation, TBO increased the protein carbonylation and lipid peroxidation levels (up to 109.40 and 119.87%, respectively) and decreased the reduced glutathione levels (59.72%) in mitochondria. TBO also uncoupled oxidative phosphorylation and photoinactivated the respiratory chain complexes I, II, and IV, as well as the FoF1-ATP synthase complex. Without photostimulation, TBO caused uncoupling of oxidative phosphorylation and loss of inner mitochondrial membrane integrity and inhibited very strongly succinate oxidase activity. TBO's uncoupling effect was clearly seen in intact livers where it stimulated oxygen consumption at concentrations of 20 and 40 μM. Additionally, TBO (40 μM) reduced cellular ATP levels (52.46%) and ATP/ADP (45.98%) and ATP/AMP (74.17%) ratios. Consequently, TBO inhibited gluconeogenesis and ureagenesis whereas it stimulated glycogenolysis and glycolysis. In conclusion, we have revealed for the first time that the efficiency of TBO as a PS may be linked to its ability to photodynamically inhibit oxidative phosphorylation. In contrast, TBO is harmful to mitochondrial energy metabolism even without photostimulation, which may lead to adverse effects when used in PDT. [ABSTRACT FROM AUTHOR]

Introduction: Sepsis-associated encephalopathy (SAE) is a diffuse cerebral dysfunction resulting from a systemic inflammatory response to infection; however, its pathophysiology remains unclear. Sepsis-induced neuroinflammation and blood-brain barrier (BBB) disruption are crucial factors in brain function disturbance in SAE. Mast cells (MCs) activation plays an important role in several neuroinflammation models; however, its role in SAE has not been comprehensively investigated. Methods: We first established a SAE model by cecal ligation puncture (CLP) surgery and checked the activation of MCs. MCs activation was checked using immumohistochemical staining and Toluidine Blue staining. We administrated cromolyn (10mg/ml), a MC stabilizer, to rescue the septic mice. Brain cytokines levels were measured using biochemical assays. BBB disruption was assessed by measuring levels of key tight-junction (TJ) proteins. Cognitive function of mice was analyzed by Y maze and open field test. Transwell cultures of brain microvascular endothelial cells (BMVECs) co-cultured with MCs were used to assess the interaction of BMVECs and MCs. Results: Results showed that MCs were overactivated in the hippocampus of CLPinduced SAE mice. Cromolyn intracerebroventricular (i.c.v) injection substantially inhibited the MCs activation and neuroinflammation responses, ameliorated BBB impairment, improved the survival rate and alleviated cognitive dysfunction in septic mice. In vitro experiments, we revealed that MCs activation increased the sensitivity of BMVECs against to lipopolysaccharide (LPS) challenge. Furthermore, we found that the histamine/histamine 1 receptor (H1R) mediated the interaction between MCs and BMVECs, and amplifies the LPS-induced inflammatory responses in BMVECs by modulating the TLR2/4-MAPK signaling pathway. Conclusions: MCs activation could mediate BBB impairment and cognitive dysfunction in septic mice in a histamine-dependent pathway. [ABSTRACT FROM AUTHOR]

Calcium phosphate forms particles under excessive urinary excretion of phosphate in the kidney. While the formation of calcium phosphate particles (CaPs) has been implicated in the damage to renal tubular cells and renal dysfunction, clarifying the ultrastructural information and the elemental composition of the small CaPs in the wide areas of kidney tissue has been technically difficult. This study introduces correlative and sequential light as well as electron microscopic CaP observation in the kidney tissue by combining fluorescent staining for CaPs and energy-dispersive X-ray spectroscopy (EDS) in scanning electron microscopy (SEM) on resin sections prepared using high-pressure freezing and freeze substitution. CaPs formed in mouse kidneys under long-term feeding of a high-phosphate diet were clearly visualized on resin sections by fluorescence-conjugated alendronate derivatives and toluidine blue metachromasia. These CaPs were verified by correlative observation with EDS. Furthermore, small CaPs formed in the kidney under short-term feeding were detected using fluorescent probes. The elemental composition of the particles, including calcium and magnesium, was identified following EDS analyses. These results suggest that the correlative microscopy approach is helpful for observing in situ distribution and elemental composition of CaPs in the kidney and contributing to studies regarding CaP formation-associated pathophysiology. [ABSTRACT FROM AUTHOR]

Background. Atopic dermatitis is a common dermatological disease, and mast cell degranulation is believed to be related with the progression of atopic dermatitis. Mas-related G protein-coupled receptor-X2 (MRGPRX2), and calcium release-activated calcium channel protein 1-2 (ORAI-1, ORAI-2) are involved in mast cell degranulation. Celastrol is an active monomer of Tripterygium wilfordii, and it presents an antiatopic role. Methods. 2,4-Dinitrofluorobenzene (DNFB) and compound 48/80 (C 48/80) were used to establish a slow and acute scratching animal model, respectively. Hematoxylin-eosin and toluidine blue staining was used to investigate tissue injury. Inflammatory factor concentration was measured with ELISA. The expression of MRGPRX2, ORAI-1, and ORAI-2 was detected with immunohistochemistry (IHC) staining. Gene expression profiling and microRNA array were performed to investigate gene differential expression. Results. Celastrol greatly inhibited atopic dermatitis-related tissues injury, mast cell production, histamine release, scratching level, inflammatory factor expression, and activation of MRGPRX2/ORAI axis in the DNFB-induced atopic dermatitis model. The influence of Celastrol on atopic dermatitis was remarkably reversed by overexpression of MRGPRX2. Conclusion. We found that the improvements of atopic dermatitis caused by Celastrol were reversed by treatment with MRGPRX2OE, indicating that Celastrol might affect atopic dermatitis through MRGPRX2. This study might provide a novel thought for the prevention and treatment of atopic dermatitis by regulating MRGPRX2. [ABSTRACT FROM AUTHOR]

The surface sulfate groups of cotton-derived cellulose nanowhiskers (CNWs) were quantified using three different methods: conductometric titration, adsorption of toluidine blue O (TBO), and sulfur elemental analysis. The former two methods indicated comparable values, whereas the last method indicated a considerable overestimation of up to 1.4 times the sulfate content compared to the values obtained by titration. This overestimation may be due to the inaccessibility of the sulfates inside the CNWs to the titrant and TBO, and/or the presence of non-sulfate sulfur elements. In addition to the quantitativeness of the TBO adsorption method, the present results also demonstrate the desulfation efficiency under several different desulfation conditions, where the sulfate content decreased by only half after alkali-mediated desulfation even with 2 M NaOH, whereas quite effective desulfation (ca. 90% decrease) was achieved with dilute HCl (0.25 M). [ABSTRACT FROM AUTHOR]

Cellulose nanofiber (CNF) derived from woody bioresources is a fascinating natural nanomaterial. In this work, spherical microparticles were fabricated by using TEMPO-oxidized CNF (TOCNF) and cellulose diacetate (CDA) via Pickering emulsion templating. The CDA-dissolved organic solvents were emulsified stably with TOCNF, followed by removing the solvents to form microspheres with core-shell structures, where the CDA cores were covered with the TOCNF shells. The prepared spherical microparticles possessed an average diameter and sphericity index of 6.4 µm and 0.96, respectively. The zeta-potential value was approximately-48 mV, indicating the stable colloidal system in water. The CDA/TOCNF microparticles were stained with toluidine blue dye for negatively-charged TOCNF. Besides, furry nanofiber-like morphology was observed on the particle surface by scanning electron microscopy. Woodderived CDA/TOCNF microspheres are a promising alternative to fossil resource-derived, non-biodegradable microbeads in cosmetic applications. [ABSTRACT FROM AUTHOR]

Numerous tools and techniques such as vital stain, autofluorescence, exfoliative cytology, histomorphometry, etc. were explored in the detection of dysplasia, but have variable sensitivity and specificity. The present study evaluated the effect of toluidine blue in increasing the accuracy of histomorphometry analysis in detection of the dysplasia in OPMDs on exfoliative cytology. An experimental, observational, cross-sectional research was performed on patients attending dental hospitals and in oral health check-up camps suspected of having OPMDs. Cytology smear was collected using the standard protocol before and after the application of a toluidine blue stain over the lesion. Two pathologists performed the dysplasia grading and histomorphometry analysis. Chi-square and t-tests were carried out (p<0.05). The study observed a positive correlation between toluidine blue positivity and dysplasia in OPMDs. The toluidine blue stain sensitivity, specificity, Positive Predictive Value, and Negative predictive value were 39.53%, 97.56, 97.14, and 43.58% respectively. Toluidine blue before the cytology smear does not improve the diagnostic accuracy of cytomorphometry. Further, there was no significant variation in cellular and nuclear morphometry parameters as compared to normal mucosa. Toluidine blue stain does not improve the efficacy of cytomorphometry in discriminating dysplasia on cytosmears. However, toluidine blue stain can assist clinicians in identifying potential areas of having a suspicion of dysplasia for scalpel biopsy. [ABSTRACT FROM AUTHOR]

In tissue engineering bioresorbable polymers are commonly used to mimetize a damaged tissue characteristics. In spite of that, we did not find a comparative study of the response of the same cell to dense, porous, and fibrous biomaterials. The present work aimed to study the in vitro of pure polymers [poly (ε-caprolactone) (PCL) and poly (hydroxybutyrate-co-hydroxyvalerate) (PHBV)] and their 75:25 blend in dense, porous, and fibrous forms in Vero cells, a fibroblast cell line. The biomaterials were characterized morphologically by scanning electron microscopy (SEM). The MTT assay was used for analysis of possible cytotoxicity. Vero cells were cultured on biomaterials for 120 h and submitted to SEM, morphological and cytochemical analyses. The dyes used were crystal violet (morphology), toluidine blue at pH 4.0 (for nucleic acids and glycosaminoglycans) and 2.5 (for glycosaminoglycans only), periodic acid-Schiff reactive (for neutral sugars), xylidine ponceau at pH 2.5 (for total proteins), and picrosirius/hematoxylin (for selective collagen analysis). The biomaterials studied were not considered citocytotoxic. The cells were able to adhere to and proliferate on the substrates. The cytochemical data indicate that the cells have high functional activity, with high protein synthesis on the scaffolds but do not accumulate acid or neutral sugars. The cells on the fibrous polymers appear to be more elongated, following the shape of the fibers. Regarding the scaffold form (porous or fibrous), the in vitro data do not show great superiority of one type to the other. Then, all the polymers tested are suitable for tissue engineering. [ABSTRACT FROM AUTHOR]

Research background. Despite clearly recognized links between increased body mass and increased risk for various pathological conditions, therapeutic options to treat obesity are still very limited. The aim of the present study is to explore the effect of low-molecular-mass collagen fragments obtained from the scales of Antarctic wild marine fish on rats' visceral and subcutaneous white adipose tissue in a high-calorie diet-induced obesity model. Experimental approach. The study was conducted on outbred rats, which were divided into 3 experimental groups: (i) control, consuming standard food (3.81 kcal/g), (ii) obese group, consuming a high-calorie diet (5.35 kcal/g), and (iii) obese group, consuming a high-calorie diet (5.35 kcal/g) with intragastric administration of low-molecular-mass collagen fragments (at a dose 1 g/kg of body mass during 6 weeks). Low-molecular-mass collagen fragments were obtained by a procedure that included collagen extraction from fish scales and enzymatic hydrolysis with pepsin. Apart from hematoxylin and eosin staining, fibrosis level was assessed by histochemical Van Gieson's trichrome picrofuchsin staining, and mast cells were analysed by toluidine blue O staining. Results and conclusions. Group treated with low-molecular-mass fragments of collagen exhibited decreased rate of mass gain, relative mass, area occupied by collagen fibre of both visceral and subcutaneous adipose tissue, and cross-sectional area of both visceral and subcutaneous adipocytes. Treatment with low-molecular-mass fragments of collagen reduced the infiltration of immune cells, number of mast cells and their redistribution back to the septa. This was also accompanied by a decreased number of the crown-like structures formed by the immune cells, which are markers of chronic inflammation that accompanies obesity. Novelty and scientific contribution. This is the first study that reports the anti-obesity effect of low-molecular-mass fragments produced as a result of controlled hydrolysis of collagen from the scales of Antarctic wild marine fish in the in vivo model. Another novelty of this work is the observation that the tested collagen fragments not only reduce the body mass, but also improve the morphological and inflammatory parameters (decrease in the number of crown-like structures, immune cell infiltration, fibrosis and mast cells). Altogether, our work suggests that low-molecular-mass collagen fragments are a promising candidate for amelioration of some comorbidities linked to obesity. [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]

Photodynamic therapy (PDT) provides an effective therapeutic option for different types of cancer in addition to surgery, radiation, and chemotherapy. The treatment outcome of PDT is largely determined by both the light and dark toxicity of photosensitizers (PSs), which can be technically improved with the assistance of a drug delivery system, especially the nanocarriers. Toluidine blue (TB) is a representative PS that demonstrates high PDT efficacy; however, its application is largely limited by the associated dark toxicity. Inspired by TB’s noncovalent binding with nucleic acids, in this study, we demonstrated that DNA nanogel (NG) could serve as an effective TB delivery vehicle to facilitate anticancer PDT. The DNA/TB NG was constructed by the simple self-assembly between TB and short DNA segments using cisplatin as a crosslinker. Compared with TB alone, DNA/TB NG displayed a controlled TB-releasing behavior, effective cellular uptake, and phototoxicity while reducing the dark toxicity in breast cancer cells MCF-7. This DNA/TB NG represented a promising strategy to improve TB-mediated PDT for cancer treatments.