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PloS One 2016The primo vascular system (PVS) is being established as a circulatory system that corresponds to acupuncture meridians. There have been two critical questions in making...
The primo vascular system (PVS) is being established as a circulatory system that corresponds to acupuncture meridians. There have been two critical questions in making the PVS accepted as a novel liquid flowing system. The first one was directly to show the flow of liquid in PVS and the second one was to explain why it was not observed in the conventional histological study of animal tissues. Flow in the PVS in the abdominal cavity was previously verified by injecting Alcian blue into a primo node. However, the tracing of the dye to other subsystems of the PVS has not been done. In the current work we injected fluorescent nanoparticles (FNPs) into a primo node and traced them along a primo vessel which was inside a fat tissue in the abdominal wall. Linea alba is a white middle line in the abdominal skin of a mammal and a band of fat tissue is located in parallel to the linea alba in the parietal side of the abdominal wall of a rat. In this fat band a primo vessel runs parallel to the prominent blood vessels in the fat band and is located just inside the parietal peritoneum. About the second question on the reason why the PVS was not in conventional histological study the current work provided the answer. Histological analysis with hematoxyline and eosine, Masson's trichrome, and Toluidine blue could not discriminate the primo vessel even when we knew the location of the PVS by the trace of the FNPs. This clearly explains why the PVS is hard to observe in conventional histology: it is not a matter of resolution but the contrast. The PVS has very similar structure to the connective tissues that surround the PVS. In the current work we propose a method to find the PVS: Observation of mast cell distribution with toluidine blue staining and the PN has a high density of mast cells, while the lymph node has low density.
Topics: Abdominal Cavity; Abdominal Fat; Abdominal Wall; Acupuncture Points; Alcian Blue; Animals; Coloring Agents; Eosine Yellowish-(YS); Hematoxylin; Humans; Lymph Nodes; Lymphatic Vessels; Male; Mast Cells; Nanoparticles; Rats; Rats, Sprague-Dawley; Rheology; Rhodamines; Staining and Labeling; Tolonium Chloride
PubMed: 26937963
DOI: 10.1371/journal.pone.0150423 -
Journal of Cytology Oct 2012Fine needle aspiration cytology (FNAC) is extensively used in the diagnosis of various clinically palpable lesions of breast and salivary glands. Much interest has been...
BACKGROUND
Fine needle aspiration cytology (FNAC) is extensively used in the diagnosis of various clinically palpable lesions of breast and salivary glands. Much interest has been gained in mucosubstances produced in tumors arising from these organs.
AIMS
To evaluate the utility of Periodic acid Schiff with diastase (PAS-D) and Alcian blue (AB) staining pattern on fine needle aspirates of breast and salivary gland neoplasms.
MATERIALS AND METHODS
Seventy-five cases of different neoplasm of breast and salivary gland were studied. The staining pattern of PAS-D and AB stains on smears of these neoplasm were observed.
RESULTS
Among cases of neoplasms of breast, intracytoplasmic PAS-D positive globules were restricted to carcinoma except in one case where PAS-D-positive globules were seen in fibroadenoma. The background substance of both mucinous carcinoma and fibroadenoma with myxoid change stained positive with PAS-D, but the pattern was different. The cases of pleomorphic adenoma and mucoepidermoid carcinoma of salivary gland showed intracytoplasmic PAS-D-positive globules. The cases of pleomorphic adenoma showed stromal positivity which was not seen in basal cell adenoma on smears.
CONCLUSION
Intracytoplasmic PAS-D-positive globules may be useful in differentiating benign and malignant lesions of breast. The presence of PAS-D positive granules are useful in differentiating various lesions of salivary glands. AB staining of stromal fragments in pleomorphic adenoma is useful in differentiating it from basal cell adenoma.
PubMed: 23326023
DOI: 10.4103/0970-9371.103938 -
Journal of Clinical Medicine Apr 2020To report on the clinico-pathological features of a series of 14 intra-oral mucoepidermoid carcinomas showing exclusive intra-cystic growth.
AIMS
To report on the clinico-pathological features of a series of 14 intra-oral mucoepidermoid carcinomas showing exclusive intra-cystic growth.
MATERIALS AND METHODS
All mucoepidermoid carcinomas diagnosed in the period 1990–2012 were retrieved; the original histological preparations were reviewed to confirm the diagnosis and from selected cases, showing exclusive intra-cystic neoplastic components, additional sections were cut at three subsequent 200 m intervals and stained with Hematoxylin–Eosin, PAS, Mucicarmine and Alcian Blue, to possibly identify tumor invasion of the adjacent tissues, which could have been overlooked in the original histological preparations. Additionally, pertinent findings collected from the clinical charts and follow-up data were analyzed.
RESULTS
We identified 14 intraoral mucoepidermoid carcinomas treated by conservative surgery and with a minimum follow up of five years. The neoplasms were located in the hard palate (nine cases), the soft palate (two), the cheek (two) and the retromolar trigone (one). In all instances, histological examination revealed the presence of a single cystic space, containing clusters of columnar, intermediate, epidermoid, clear and mucous-producing cells, the latter exhibiting distinct intra-cytoplasmic mucin production, as confirmed by PAS, Mucicarmine and Alcian Blue stains. The cysts were entirely circumscribed by fibrous connective tissue, and no solid areas or infiltrating tumor cell clusters were detected. Conservative surgical resection was performed in all cases, and no recurrences or nodal metastases were observed during follow up.
CONCLUSIONS
Mucoepidermoid carcinomas showing prominent (>20%) intra-cystic proliferation currently are considered low-grade tumors. In addition, we also unveil the possibility that mucoepidermoid carcinomas, at least in their early growth phase, may display an exclusive intra-cystic component and might be considered as in situ carcinomas, unable to infiltrate adjacent tissues and metastasize.
PubMed: 32325647
DOI: 10.3390/jcm9041157 -
Regenerative Biomaterials 2022Autologous chondrocytes (C cells) are effective sources of cell therapy for engineering cartilage tissue to repair chondral defects, such as degenerative arthritis. The...
Autologous chondrocytes (C cells) are effective sources of cell therapy for engineering cartilage tissue to repair chondral defects, such as degenerative arthritis. The expansion of cells with C cell characteristics has become a major challenge due to inadequate donor sites and poor proliferation of mature C cells. The perichondrial progenitor cells (P cells) from the cambium layer of the perichondrium possessed significantly higher mesenchymal stem cell markers than C cells. In the transwell co-culture system, P cells increased the passaging capacity of C cells from P6 to P9, and the cell number increased 128 times. This system increased the percentage of Alcian blue-positive C cells from 40% in P6 to 62% in P9, contributing about 198 times more Alcian blue-positive C cells than the control group. C cells co-cultured with P cells also exhibited higher proliferation than C cells cultured with P cell-conditioned medium. Similar results were obtained in nude mice that were subcutaneously implanted with C cells, P cells or a mixture of the two cell types, in which the presence of both cells enhanced neocartilage formation . In aggregate, P cells enhanced the proliferation of C cells in a dose-dependent manner and prolonged the longevity of mature C cells for clinical applications.
PubMed: 35702349
DOI: 10.1093/rb/rbab078 -
Ocular Oncology and Pathology Apr 2018To investigate the source of fibrous astrocytes and neuroblasts in a small ciliary body medulloepithelioma appearing as a leukocoria in a 3-week-old baby girl.
PURPOSE
To investigate the source of fibrous astrocytes and neuroblasts in a small ciliary body medulloepithelioma appearing as a leukocoria in a 3-week-old baby girl.
METHODS
Histopathologic and immunohistochemical studies included Alcian blue, periodic acid-Schiff, and antisera for the detection of S100 protein, CD99, glial fibrillary acidic protein (GFAP), CRX, NeuN, neurofilaments, synaptophysin, desmin, and myogenin.
RESULTS
A small, nonteratoid ciliary body medulloepithelioma with collections of Alcian blue+ mucoplysaccharides was present in the enucleated globe. The retinal mass displayed multilaminar dysplastic rosettes that were CRX+, NeuN-, and synaptophysin-. Intraretinal neurofilaments and scattered NeuN+ neurocytes were also identified. At the base of the retinal mass ribbons and pseudopapillae of CRX+, NeuN- medullary epithelium were found. The latter developed from an S100+ and weakly CD99+ monolayer of premedullary epithelium. GFAP+ fibrous astrocytes and NeuN- neuroblasts streamed from the medullary epithelium.
CONCLUSIONS
A multilaminar medullary epithelium and a precursor monolayer of premedullary epithelium were both identified. Neuroblasts and fibrous astrocytes were determined to arise separately from the medullary epithelium. The early stage of tumorigenesis afforded by a small tumor provided the opportunity to discover morphologic and immunohistochemical evidence for these differentiations.
PubMed: 29765950
DOI: 10.1159/000481287 -
Molecules (Basel, Switzerland) Feb 2018In acidic media, cationic phthalocyanine Alcian blue 8GX, has an efficient fluorescence quenching effect on anionic phthalocyanine tetrasulphoaluminium phthalocyanines...
In acidic media, cationic phthalocyanine Alcian blue 8GX, has an efficient fluorescence quenching effect on anionic phthalocyanine tetrasulphoaluminium phthalocyanines (AlS₄Pc), forming an almost non-fluorescent associate. Based on this discovery, a red-emitting fluorescent probe consisted of AlS₄P and Alcian blue 8GX has been developed through molecular assembly. Further studies indicated that the presence of Hg(II) ion has a significant fluorescence recovery effect of the probe. Notably, only Hg(II) can significantly restore the fluorescence of AlS₄Pc-Alcian blue 8GX system which was revealed from the screening experiments of common metal ions, which confirmed that the fluorescence recovery by other metal ions is very weak or even unrestored, showing high specificity and sensitivity AlS₄Pc-Alcian blue 8GX to Hg(II). Thus, a new fluorimetry for Hg(II) with high specificity and high sensitivity in a wide concentration range has been established using AlS₄P-Alcian blue 8GX associate as a red-emitting fluorescent probe. It is more noteworthy that this study opens a new way for development and application of functional phthalocyanine based red-emitting fluorescent probes.
Topics: Alcian Blue; Biosensing Techniques; Fluorescence; Fluorescent Dyes; Indoles; Isoindoles; Mercury; Spectrometry, Fluorescence
PubMed: 29443913
DOI: 10.3390/molecules23020418 -
Journal of Clinical and Diagnostic... Jan 2014Incidence of prostate cancer in India is relatively low compared to the western countries. Nevertheless, an increase by 1% yearly has been recorded in the last three...
BACKGROUND
Incidence of prostate cancer in India is relatively low compared to the western countries. Nevertheless, an increase by 1% yearly has been recorded in the last three years, thereby making early diagnosis of prostate cancer crucial for controlling its incidence. Differentiating between benign and malignant lesions has been a diagnostic dilemma, especially in prostate pathology. This is compounded by unavailability of modern tests in certain regions of developing nations.
METHODS
A cohort of one hundred seventy six prostatomegaly patients used in the current study was obtained both retrospectively and prospectively at the Jawaharlal Nehru Medical College, Sawangi, Wardha, Maharashtra, India. Details of the patients were recorded which included their age. The samples were then cut into 5 sections, each of 5micron thickness. One section was preserved and the other 4 sections were subjected to Hematoxylin and Eosin (H and E), Periodic Acid-Schiff (PAS), Alcian Blue and AgNOR stains. Degree of differentiation was estimated and correlated with the Gleason score and the outcome of the stainings.
RESULTS
Majority of benign prostatic hyperplasia and all primary carcinoma patients were in their sixth to eighth decade of life. While all the benign lesions were negative, 6 out of 9 primary prostate carcinomas were positive for Alcian Blue stain. Majority of both benign and malignant lesions were positive for Periodic Acid Schiff (PAS) stain. In terms of Argyrophilic Nucleolar Organiser Region (AgNOR) count per nucleus, the value in benign lesions was observed to be half the count observed in malignant lesions per nucleus.
CONCLUSION
Although the potential use of the orthodox stains individually may not serve the purpose to differentiate between benign and malignant lesions, together they may have the potential to identify relatively more malignant cases. This may be helpful especially in low socio-economic countries and rural areas where molecular based tests may not yet be available.
PubMed: 24596742
DOI: 10.7860/JCDR/2014/7002.3938 -
Medical Molecular Morphology Jun 2021Glycocalyx (GCX) is a thin layer of negatively charged glycoproteins that covers the vascular endothelial surface and regulates various biological processes. Because of...
Glycocalyx (GCX) is a thin layer of negatively charged glycoproteins that covers the vascular endothelial surface and regulates various biological processes. Because of the delicate and fragile properties of this structure, it is difficult to detect GCX morphologically. We established a simple method for a three-dimensional visualization of endothelial GCX using low-vacuum scanning electron microscopy (LVSEM) on formalin-fixed paraffin-embedded (FFPE) sections. Mouse kidney tissue was fixed with 10% buffered formalin containing 1% Alcian blue (ALB) via perfusion and immersion. FFPE sections were observed by light microscopy (LM) and LVSEM, and formalin-fixed epoxy resin-embedded ultrathin sections were observed by transmission electron microscopy (TEM). The endothelial GCX from various levels of kidney blood vessels was stained blue in LM and confirmed as a thin osmiophilic layer in TEM. In LVSEM, the sections stained by periodic acid methenamine silver (PAM) revealed the endothelial GCX as a layer of dense silver-enhanced particles, in both the samples fixed via perfusion and immersion. Correlative light and electron microscopy (CLEM) revealed the fine visible structure of endothelial GCX. This simple method using FFPE samples with ALB will enable the three-dimensional evaluation of endothelial GCX alterations in various human diseases associated with endothelial injury in future studies.
Topics: Alcian Blue; Animals; Endothelial Cells; Female; Glycocalyx; Mice; Mice, Inbred BALB C; Microscopy, Electron, Scanning; Microscopy, Electron, Transmission; Silver
PubMed: 33025157
DOI: 10.1007/s00795-020-00267-1 -
Cornea Apr 2013To investigate the normal palpebral conjunctival histology in C57BL/6 mice and the structural changes that occur in a dry eye model.
PURPOSE
To investigate the normal palpebral conjunctival histology in C57BL/6 mice and the structural changes that occur in a dry eye model.
METHODS
Twenty-four male and female C57BL/6 mice, 8 untreated and 16 exposed to experimental ocular surface desiccating stress (DS). Ocular dryness was induced by administration of scopolamine hydrobromide (0.5 mg/0.2 mL) four times a day for 5 days (DS5) or 10 days (DS10). Counts and measurements were obtained using anatomical reference points, and goblet cell density was investigated with a variety of stains.
RESULTS
Near the junction between the lid margin and the normal palpebral conjunctiva, the epithelium had an average thickness of 45.6 ± 10.5 μm, 8.8 ± 2.0 cell layers, versus 37.7 ± 5.6 μm, 7.4 ± 1.3 layers in DS10 (P < 0.05). In the goblet cell-populated palpebral region, the normal epithelium was thicker (P < 0.05) than on DS5 and DS10. In the control, 43% of the goblet cells were covered by squamous epithelium compared with 58% (DS5) and 63% (DS10) (P < 0.05). A decreased number of periodic acid-Schiff (PAS)-stained goblet cells and Alcian blue-stained goblet cells were observed in the dry eye. Not all goblet cells were stained with PAS and Alcian blue.
CONCLUSION
The mouse palpebral conjunctival epithelium was structurally similar to the human. After DS, the palpebral conjunctival epithelium decreased in thickness and goblet cell access to the surface seemed to be inhibited by surrounding epithelial cells, potentially slowing down their migration to the surface. Differential staining with PAS and Alcian blue suggests that there may be different subtypes of conjunctival goblet cells.
Topics: Analysis of Variance; Animals; Conjunctiva; Disease Models, Animal; Dry Eye Syndromes; Epithelial Cells; Female; Immunohistochemistry; Male; Mice; Mice, Inbred C57BL
PubMed: 23146932
DOI: 10.1097/ICO.0b013e318265682c -
Current Biology : CB Jun 2022The kinetochore links chromosomes to spindle microtubules to drive chromosome segregation at cell division. While we know nearly all mammalian kinetochore proteins, how...
The kinetochore links chromosomes to spindle microtubules to drive chromosome segregation at cell division. While we know nearly all mammalian kinetochore proteins, how these give rise to the strong yet dynamic microtubule attachments required for function remains poorly understood. Here, we focus on the Astrin-SKAP complex, which localizes to bioriented kinetochores and is essential for chromosome segregation but whose mechanical role is unclear. Live imaging reveals that SKAP depletion dampens the movement and decreases the coordination of metaphase sister kinetochores and increases the tension between them. Using laser ablation to isolate kinetochores bound to polymerizing versus depolymerizing microtubules, we show that without SKAP, kinetochores move slower on both polymerizing and depolymerizing microtubules and that more force is needed to rescue microtubules to polymerize. Thus, in contrast to the previously described kinetochore proteins that increase the grip on microtubules under force, Astrin-SKAP reduces the grip, increasing attachment dynamics and force responsiveness and reducing friction. Together, our findings suggest a model where the Astrin-SKAP complex effectively "lubricates" correct, bioriented attachments to help preserve them.
Topics: Alcian Blue; Animals; Cell Cycle Proteins; Chromosome Segregation; Friction; Kinetochores; Mammals; Microtubule-Associated Proteins; Microtubules; Mitosis; Phenazines; Phenothiazines; Resorcinols; Spindle Apparatus
PubMed: 35580605
DOI: 10.1016/j.cub.2022.04.061