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International Journal of Molecular... Dec 2023Alzheimer's disease (AD) is a neurodegenerative disorder characterized by the accumulation of amyloid‑β (Aβ) in the brain. The gut/brain axis may serve a role in AD...
Alzheimer's disease (AD) is a neurodegenerative disorder characterized by the accumulation of amyloid‑β (Aβ) in the brain. The gut/brain axis may serve a role in AD pathogenesis. The present study investigated deposition of Aβ in the intestinal epithelium and its potential effects on intestinal barrier function in a transgenic mouse model of AD. To investigate alterations in the structure and functionality of the intestinal mucosal barrier in AD model mice, hematoxylin and eosin staining for Paneth cell count, Alcian blue‑periodic acid Schiff staining for goblet cells, immunohistochemistry and immunofluorescence for mucin (MUC)2 and wheat germ agglutin expression, transmission electron microscopy for mucosal ultrastructure, FITC‑labeled dextran assay for intestinal permeability, quantitative PCR for goblet cell precursor expression and western blot analysis for tight junction proteins, MUC2 and inflammatory cytokine detection were performed. The results showed that AD model mice exhibited excessive Aβ deposition in the intestinal epithelium, which was accompanied by increased intestinal permeability, inflammatory changes and decreased expression of tight junction proteins. These alterations in the intestinal barrier led to an increased proliferation of goblet and Paneth cells and increased mucus synthesis. Dysfunction of gut barrier occurs in AD and may contribute to its etiology. Future therapeutic strategies to reverse AD pathology may involve early manipulation of gut physiology and its microbiota.
Topics: Mice; Animals; Mucins; Alzheimer Disease; Tight Junctions; Intestinal Mucosa; Mice, Transgenic; Permeability; Tight Junction Proteins
PubMed: 37830152
DOI: 10.3892/ijmm.2023.5316 -
Molecules (Basel, Switzerland) Oct 2020The present study aims to investigate the immunomodulatory effects of essential oil from (EOCO) in an ovalbumin (OVA)-induced allergic rhinitis (AR) mouse model. BALB/c...
The present study aims to investigate the immunomodulatory effects of essential oil from (EOCO) in an ovalbumin (OVA)-induced allergic rhinitis (AR) mouse model. BALB/c mice were intraperitoneally sensitized and stimulated with OVA. From day 22 to 35, 0.01% and 0.1% ECOC was intranasally administered 1 h before OVA stimulation. Nasal symptoms, as well as serum total and OVA-specific immunoglobulin (Ig) E levels, were measured. Interleukin (IL)-4, IL-10, interferon (IFN)-γ, and tumor necrosis factor (TNF)-α levels in nasal lavage fluid (NLF) and their production by activated splenocytes were measured. Histological changes in the sinonasal mucosa were evaluated through hematoxylin and eosin and periodic acid-Schiff staining procedure. Th cytokines and their transcription factor mRNA expressions were determined using reverse-transcription polymerase chain reaction. Intranasal EOCO administration significantly suppressed allergic symptoms, OVA-specific IgE level, sinonasal mucosal inflammatory cell infiltration, and mucus-producing periodic acid-Schiff (PAS) positive cell count. EOCO also significantly inhibited IL-4, IL-10, and TNF-α levels in NLF and activated splenocytes. Th2 and Treg related cytokines and their transcription factors in sinonasal mucosa were significantly suppressed through intransal EOCO instillation. In conclusion, repetitive EOCO intranasal instillation showed anti-inflammatory and anti-allergic effects by suppressing nasal symptoms and inhibiting the production and expression of inflammatory mediators in the OVA-induced AR mouse model.
Topics: Animals; Chamaecyparis; Disease Models, Animal; Female; Immunoglobulin E; Immunologic Factors; Inflammation Mediators; Mice, Inbred BALB C; Nasal Lavage Fluid; Nasal Mucosa; Oils, Volatile; Ovalbumin; Rhinitis, Allergic; Spleen; Transcription Factors
PubMed: 33023110
DOI: 10.3390/molecules25194517 -
Frontiers in Veterinary Science 2022In this study, we isolated and cultured canine and feline 3D corneal organoids. Samples derived from corneal limbal epithelium from one canine and one feline patient...
In this study, we isolated and cultured canine and feline 3D corneal organoids. Samples derived from corneal limbal epithelium from one canine and one feline patient were obtained by enucleation after euthanasia. Stem cell isolation and organoid culture were performed by culturing organoids in Matrigel. Organoids were subsequently embedded in paraffin for further characterization. The expression of key corneal epithelial and stromal cell markers in canine and feline organoids was evaluated at the mRNA level by RNA-ISH and at the protein level by immunofluorescence (IF) and immunohistochemistry (IHC), while histochemical analysis was performed on both tissues and organoids using periodic-acid Schiff (PAS), Sirius Red, Gomori's Trichrome, and Colloidal Iron stains. IF showed consistent expression of AQP1 within canine and feline organoids and tissues. P63 was present in canine tissues, canine organoids, and feline tissues, but not in feline organoids. Results from IHC staining further confirmed the primarily epithelial origin of the organoids. Canine and feline 3D corneal organoids can successfully be cultured and maintained and express epithelial and stem cell progenitor markers typical of the cornea. This novel model can be used in veterinary ophthalmology disease modeling, corneal drug testing, and regenerative medicine.
PubMed: 36406087
DOI: 10.3389/fvets.2022.1050467 -
Indian Journal of Dermatology,... 2020Diagnosis of onychomycosis involves direct microscopic examination with potassium hydroxide, culture or histopathology with periodic acid-Schiff staining. Nail... (Comparative Study)
Comparative Study Observational Study
BACKGROUND
Diagnosis of onychomycosis involves direct microscopic examination with potassium hydroxide, culture or histopathology with periodic acid-Schiff staining. Nail dermoscopy (onychoscopy) is a noninvasive, rapid and easily available diagnostic tool though its utility in onychomycosis remains unexplored.
OBJECTIVE
To describe the various onychoscopic patterns and compare its percentage positivity with that of standard potassium hydroxide examination, culture and histopathology in patients with a clinical diagnosis of onychomycosis.
METHODS
The study recruited 100 patients with a presumptive clinical diagnosis of onychomycosis. A detailed history, physical examination including that of nails and clinical photography was followed by onychoscopy with DermLite DL3. The nail clippings were sent for direct microscopic examination with potassium hydroxide, mycological culture and histopathology with periodic acid-Schiff stain. The patient was said to have onychomycosis if at least one of the three tests was positive.
RESULTS
Onychomycosis was confirmed by potassium hydroxide and/or culture and/or histopathology in 88 patients. Onychoscopic features were identified and their association with different clinical variants of onychomycosis was attempted. Percentage positivity for diagnosing onychomycosis in decreasing order was: direct microscopic examination with potassium hydroxide followed by spiked pattern, subungual hyperkeratosis, distal irregular termination on onychoscopy, histopathology, mycological culture and ruins aspect again observed on onychoscopy.
LIMITATIONS
Small sample size.
CONCLUSIONS
Many onychoscopic features are highly specific for different variants of onychomycosis so onychoscopy may serve as an important and quick adjunct to diagnose onychomycosis until other time-consuming investigations, such as culture and periodic acid-Schiff become available. Studies on a larger population will help arrive at a logistic conclusion.
Topics: Adult; Colony Count, Microbial; Cross-Sectional Studies; Dermoscopy; Humans; Hydroxides; Microscopy; Onychomycosis; Potassium Compounds; Predictive Value of Tests
PubMed: 32415047
DOI: 10.4103/ijdvl.IJDVL_100_19 -
Folia Morphologica 2022Glaucoma is a group of eye diseases that can cause vision loss. Prostaglandin analogues (PGAs) are known to be first-line treatment for patients with glaucoma....
BACKGROUND
Glaucoma is a group of eye diseases that can cause vision loss. Prostaglandin analogues (PGAs) are known to be first-line treatment for patients with glaucoma. Latanoprost is a good, efficient and well-tolerated PGA that is currently available as latanoprost with benzalkonium chloride (BAC) (Xalatan) and preservative-free (PF) prostaglandin analogue latanoprost (Monopost). Lately, using PF anti-glaucoma agents has been considered an essential procedure for enhancing glaucoma care. This study aimed to analyse the histological changes within the corneal tissue with the use of currently available preserved prostaglandins-derived eye drops and PF prostaglandin analogue.
MATERIALS AND METHODS
In this study, 40 male guinea pigs were divided into four equal groups. Control group, Latanoprost with 0.02% BAC-treated group, Recovery group and PF latanoprost-treated group. After 2 months, the corneal tissues of guinea pigs were prepared for light and electron microscopic studies; morphometric and statistical studies were performed.
RESULTS
Our results indicated that guinea pigs treated with latanoprost with BAC exhibited ocular surface changes: there was epithelial thinning with desquamation, the stroma showed irregularly arranged collagen fibres and small keratocytes. Morphometrically, there was a marked decrease in the thickness of epithelium and number of keratocytes. Negative periodic acid Schiff (PAS) reaction was observed in some parts of the epithelial basement membrane. The epithelium gave a strong positive immunoreactivity for Bcl-2-associated X protein (BAX). Guinea pigs left to recover exhibited improvement, while treatment of animals with PF latanoprost resulted in nearly normal corneal structure.
CONCLUSIONS
In conclusion, PF prostaglandin anti-glaucoma medication seems to be better and have protective effect on cornea of male guinea pigs than prostaglandins with BAC preservative.
Topics: Animals; Antihypertensive Agents; Cornea; Guinea Pigs; Humans; Latanoprost; Male; Preservatives, Pharmaceutical; Prostaglandins, Synthetic
PubMed: 33438192
DOI: 10.5603/FM.a2020.0146 -
Frontiers in Microbiology 2022Histological staining methods for identification vary in accuracy. This study aimed to investigate the clinical value of Grocott methenamine silver (GMS), periodic...
BACKGROUND
Histological staining methods for identification vary in accuracy. This study aimed to investigate the clinical value of Grocott methenamine silver (GMS), periodic acid-Schiff (PAS), and Alcian blue (AB) staining in the diagnosis of pulmonary cryptococcosis (PC).
METHODS
From April 2004 to June 2021, the clinical and pathological data of 152 patients with PC were collected from the Department of Pathology, Sun Yat-sen University Cancer Center. The sensitivity and identifiability of GMS, PAS, and AB staining for histological diagnosis were systematically evaluated using statistical methods combined with the microscopic characteristics of PC cases.
RESULTS
Statistical analysis showed that the detection rates of GMS, PAS, and AB staining were 100.0% (152/152), 94.7% (144/152), and 81.6% (124/152), respectively. McNemar's test showed that the sensitivity of GMS was significantly higher than those of PAS ( = 0.008) and AB stains ( < 0.001). Both PAS and AB stains had obvious non-specific staining, which interfered with the detection of , and increased diagnostic difficulties. In contrast, in GMS staining, spores were prominent with a clean background and were clearly observed at low or medium power magnification, with the identifiability significantly better than those of PAS or AB staining.
CONCLUSION
GMS staining had sensitivity up to 100%, and identifiability that was better than those of PAS and AB staining. GMS is the best method for histological diagnosis of PC.
PubMed: 35572658
DOI: 10.3389/fmicb.2022.885511 -
Journal of Clinical and Experimental... Jan 2023Sporotrichosis is an uncommon subacute or chronic infection caused by Sporothrix spp. In some urban areas of Latin America, sporotrichosis has been considered an...
Sporotrichosis is an uncommon subacute or chronic infection caused by Sporothrix spp. In some urban areas of Latin America, sporotrichosis has been considered an emergent cosmopolitan disease of zoonotic transmission by domestic cats. There are four different clinical forms of the disease: fixed cutaneous, lymphocutaneous, multifocal or disseminated cutaneous, and extracutaneous. The oral mucosa is rarely involved, usually as unspecified chronic ulcers in the context of multifocal or disseminated cutaneous form of systemic sporotrichosis. Microscopical features include chronic granulomatous inflammation containing microabscesses and fungal hyphae positive for Periodic acid Schiff and silver-based stains. The diagnosis of sporotrichosis is usually based on culture detection and strict correlation of clinical, microscopical and laboratorial data. We herein contribute with two additional illustrative cases of oral manifestation of sporotrichosis in immunocompromised patients from an endemic urban area from Rio de Janeiro-Brazil. Sporotrichosis, ulcer, oral cavity, immunosuppression.
PubMed: 36755684
DOI: 10.4317/jced.59040 -
International Journal of Biological... Dec 2020Bioadhesives have a potential to modulate the wound closure process with significant biological outcomes. However, none of the currently commercialized adhesives are...
Bioadhesives have a potential to modulate the wound closure process with significant biological outcomes. However, none of the currently commercialized adhesives are satisfactory in their performance. It is a challenging task to develop an adhesive system that can work on wet surface and enhances tissue repair and closure. In this study, we have fabricated a series of gelatin-dopamine (Gel-dop) conjugates and studied their adhesive properties after being chemically crosslinked using sodium periodate. The designed material was assessed for its adhesive properties including tensile, lap shear and peeling study by varying the degree of dopamine substitution. It was observed that the adhesive property has a direct correlation with increase in dopamine content until reaching a maximum and then a subsequent decrease. We tested the adhesive strength of the different formulations by varying the degree of substitution and compared against fibrin glue, which is considered as the gold standard of adhesives. The formulation with a moderate substitution degree demonstrated the optimal adhesive property than those formulations with lower and larger substitution degree. Further, the in vitro cytotoxicity study showed that this tunable Gel-dop adhesives are to non-cytotoxic, indicating a potential use in clinic applications. This study illustrates that adhesiveness can be regulated by changing the degree of dopamine substitution.
Topics: Adhesiveness; Animals; Benzoquinones; Catechols; Cell Adhesion; Cell Survival; Cross-Linking Reagents; Dopamine; Fibrin Tissue Adhesive; Gelatin; Hydrogels; Materials Testing; Oxygen; Periodic Acid; Pressure; Rheology; Shear Strength; Skin; Surface Properties; Swine; Tensile Strength; Tissue Adhesives
PubMed: 32721461
DOI: 10.1016/j.ijbiomac.2020.07.195 -
Toxicological Research Jan 2022This study was performed to provide information on classifying eye irritating chemicals using the BCOP assay. After the BCOP assay, bovine corneas were classified by...
This study was performed to provide information on classifying eye irritating chemicals using the BCOP assay. After the BCOP assay, bovine corneas were classified by IVIS presented in OECD test guideline 437, and three special staining methods (H&E, MT, and PAS) were performed for histopathological evaluation. Non-irritant chemicals (IVIS ≤ 3), showed intact structures. In the 3 < IVIS ≤ 55 group, epithelial cell edema was observed by H&E staining, and loose collagen bundles were confirmed by MT staining. In PAS staining, bleaching of the epithelium and reduced visibility of the basement membrane were observed. Severe irritant chemicals (IVIS > 55) showed large increases edema and nuclear condensation by H&E staining. Loose collagen bundles and vacuoles around keratocytes were also observed by MT staining. Bleaching of the epithelial layer, reduction in visibility, and thickness of the basement membrane were confirmed by PAS staining. Based on the stepwise histopathological analysis, we set the criteria and grades for histopathological evaluation and found that eye irritation was increased following the irritation degree of test chemicals. Further histopathological study will support and lead to improvements in the BCOP assay.
PubMed: 35070936
DOI: 10.1007/s43188-021-00099-w -
Frontiers in Plant Science 2022Seedlessness is one of the most important agronomic traits in mandarins on the fresh fruit market. Creation of triploid plants is an important breeding strategy for...
Seedlessness is one of the most important agronomic traits in mandarins on the fresh fruit market. Creation of triploid plants is an important breeding strategy for development of new commercial varieties of seedless citrus. To this end, one strategy is to perform sexual hybridizations, with tetraploid genotypes as male parents. However, while seed development has been widely studied in citrus, knowledge of key steps such as microsporogenesis and microgametogenesis, is scarce, especially in polyploids. Therefore, we performed a study on the effect of ploidy level on pollen development by including diploid and tetraploid (double diploid) genotypes with different degrees of pollen performance. A comprehensive study on the pollen ontogeny of diploid and doubled diploid "Sanguinelli" blood orange and "Clemenules" clementine was performed, with focus on pollen grain germination and , morphology of mature pollen grains by scanning electron microscopy (SEM), cytochemical characterization of carbohydrates by periodic acid-Shiff staining, and specific cell wall components revealed by immunolocalization. During microsporogenesis, the main difference between diploid and doubled diploid genotypes was cell area, which was larger in doubled diploid genotypes. However, after increase in size and vacuolization of microspores, but before mitosis I, doubled diploid "Clemenules" clementine showed drastic differences in shape, cell area, and starch hydrolysis, which resulted in shrinkage of pollen grains. The loss of fertility in doubled diploid "Clemenules" clementine is mainly due to lack of carbohydrate accumulation in pollen during microgametogenesis, especially starch content, which led to pollen grain abortion. All these changes make the pollen of this genotype unviable and very difficult to use as a male parent in sexual hybridization with the objective of recovering large progenies of triploid hybrids.
PubMed: 35557738
DOI: 10.3389/fpls.2022.862813