Rapid detection of biofilm with modified alcian blue staining: In‐vitro protocol improvement and validation with clinical cases

2020 ◽  
Vol 28 (6) ◽  
pp. 834-843
Author(s):  
Yu‐Feng Wu ◽  
Tyng‐Yuh Lee ◽  
Wan‐Ting Liao ◽  
Ho‐Hsien Chuan ◽  
Nai‐Chen Cheng ◽  
...  
Keyword(s):  
Alcian Blue ◽  
Rapid Detection ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Clinical Cases

scholarly journals Human osteogenic protein-1 induces both chondroblastic and osteoblastic differentiation of osteoprogenitor cells derived from newborn rat calvaria.

1993 ◽  
Vol 123 (4) ◽  
pp. 921-933 ◽  
Author(s):  
I Asahina ◽  
T K Sampath ◽  
I Nishimura ◽  
P V Hauschka
Keyword(s):  
Alkaline Phosphatase ◽  
Alcian Blue ◽  
Osteoblastic Differentiation ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Tgf Beta ◽  
Newborn Rat ◽  
Osteoprogenitor Cells ◽  
Osteogenic Protein

Osteogenetic protein-1 (OP-1), a member of the TGF-beta superfamily, induces endochondrial bone formation at subcutaneous sites in vivo and stimulates osteoblastic phenotypic expression in vitro. Primary cultures of newborn rat calvarial cells contain a spectrum of osteogenic phenotypes ranging from undifferentiated mesenchymal osteoprogenitor cells to parathyroid hormone (PTH)-responsive osteoblasts. We examined whether treatment of this cell population with recombinant human osteogenic protein-1 could induce chondrogenesis in vitro. Markers of chondroblastic versus osteoblastic differentiation included alcian blue staining at pH 1, alkaline phosphatase-specific activity, osteocalcin radioimmunoassay, and expression of collagen mRNAs. 6 d of treatment (culture days 1-7) with 4-100 ng OP-1/ml caused dose-dependent increases in alcian blue staining intensity and alkaline phosphatase activity (4.7- and 3.4-fold, respectively, at 40 ng/ml), while osteocalcin production decreased twofold. Clusters of round, refractile, alcian blue-stained cells appeared by day 3, increased in number until day 7, and then became hypertrophic and gradually became less distinct. Histochemically, the day 7 clusters were associated with high alkaline phosphatase activity and became mineralized. mRNA transcripts for collagen types II and IX were increased by OP-1, peaking at day 4, while type X collagen mRNA was detectable only on day 7 in OP-1-treated cultures. Delay of OP-1 exposure until confluence (day 7) amplifies expression of the normal osteoblastic phenotype and accelerates its developmental maturation. In contrast, early OP-1 treatment commencing on day 1 strongly amplifies chondroblastic differentiation. In the same protocol, TGF-beta 1 alone at 0.01-40 ng/ml fails to induce any hypertrophic chondrocytes, and in combination with OP-1, TGF-beta 1 blocks OP-1-dependent chondroinduction. OP-1 is believed to act on a subpopulation of primitive osteoprogenitor cells to induce endochondrial ossification, but does not appear to reverse committed osteoblasts to the chondrocyte phenotype.

Antibiotic impregnated orthopaedic bone cement: Kinetics and impact of teicoplanin on staphylococcal colonization

1994 ◽  
Vol 52 ◽  
pp. 482-483
Author(s):  
Charles E. Edmiston ◽  
Michael P. Goheen
Keyword(s):  
Staphylococcus Aureus ◽  
Bone Cement ◽  
Alcian Blue ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Orthopaedic Implant ◽  
Glycopeptide Antibiotic ◽  
Colony Forming Units ◽  
Drug Free

Previous studies in our laboratory have questioned the efficacy of antibiotic impregnated bone cement to prevent staphylococcal colonization of orthopaedic implants following surgery. Teicoplanin a glycopeptide antibiotic with excellent anti-staphylococcal activity was added to polymethylmethacrylate in concentrations ranging from 2-5 wt%. Five staphylococcal strains recovered from orthopaedic implant infections were chosen for study: 2-Staphylococcus aureus and 3-Staphylococcus eoidermidis. Two of the staphylococcal isolates were identified as producing an exopolysaccharide slime following alcian blue staining with lysine. The in vitro release of antibiotic from the resin surface was determined by biological assay at 0, 12, 24, 48, 96 and 10 days post-study. Mean teicoplanin concentrations ranged from 200 to 430 ug/ml at 0 hour and 22 to 50 ug/ml at 10 days post-study. Teicoplanin impregnated and drug free resin controls were exposed to standardized inoculum (5.0 log10 colony forming units/ml) of the staphylococcal clinical isolates and incubated at 35°C in physiologic buffered saline with 0.25% dextrose.

Spontaneous release of submucosal gland serous and mucous cell macromolecules from human nasal explants in vitro

1996 ◽  
Vol 270 (4) ◽  
pp. L595-L600
Author(s):  
M. Ali ◽  
J. Maniscalco ◽  
J. N. Baraniuk
Keyword(s):  
Alcian Blue ◽  
Mucous Cell ◽  
Blue Staining ◽  
Secretory Cells ◽  
Alcian Blue Staining ◽  
Serous Cell ◽  
Respiratory Cells ◽  
Mucosal Explants ◽  
Respiratory Epithelial

Respiratory epithelial and gland cells cultured in vitro demonstrate changes from differentiated serous and mucous cells toward intermediate "seromucous" cells. This spontaneous process was examined by culturing human nasal mucosal explants in CMRL 1066 medium without growth factors for 6 days and measuring the concentrations of spontaneously released serous cell products [lactoferrin, lysozyme, 7F10-immunoreactive mucoglycoconjugates (7F10-irm)] and Alcian blue-staining mucous cell products. 7F10-irm was progressively and significantly increased on each day of culture. In contrast, lysozyme, lactoferrin, and Alcian blue-staining material decreased significantly. Each had its own pattern of decreasing release. Dexamethasone (1 microM) had no effect on these trends. Phorbol myristate ester (PMA; 100 nM) reduced 7F10-irm release on days 4-6 and delayed the drop in lactoferrin release. Dexamethasone blunted these effects of PMA. These data indicate that respiratory secretory cells alter their phenotypes when cultured in vitro and progressively change the relative amounts of mucoglycoconjugates and proteins spontaneously released. These changes should be anticipated when interpreting experiments involving cultured respiratory cells.

THU0055 ANTI-DEGRADATIVE AND PRO-CHONDROGENIC PROPERTIES OF LIRAGLUTIDE, A GLUCAGON-LIKE-PEPTIDE 1 RECEPTOR AGONIST: EVIDENCE FROM PRECLINICAL STUDIES AND IMPLICATION FOR OSTEOARTHRITIS

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 239.1-239
Author(s):  
F. Berenbaum ◽  
C. Meurot ◽  
J. Breton ◽  
L. Sudre ◽  
C. Bougault ◽  
...  
Keyword(s):  
Alcian Blue ◽  
Articular Chondrocytes ◽  
Inflammatory Responses ◽  
Joint Disease ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Glucagon Like Peptide ◽  
Glucagon Like Peptide 1

Background:Osteoarthritis (OA) is a degenerative joint disease affecting millions of individuals worldwide. Its development has been reported to be associated with cartilage degradation and inflammatory responses leading to pain, swelling and reduced function. Although OA is a disorder of the whole joint, the progressive destruction of cartilage extracellular matrix is considered as its hallmark. To date, approved OA treatments are only symptomatic. Therefore, there is an urgent need to explore disease-modifying OA drugs (DMOADs) that can mitigate, stop, or even reverse the development of OA.Objectives:In this context, the objective of this study was to assess the effect of liraglutide, a Glucagon-Like-Peptide 1 Receptor (GLP-1R) agonist approved for type 2 diabetes, on chondrogenesis, catabolism/inflammation and cartilage protection inin vitroandin vivopreclinical models of OA.Methods:The capacity of liraglutide to induce chondrogenesis was evaluated using primary human mesenchymal stem cells (hMSCs). Alcian blue staining was used to assess differentiation of hMSC into chondrocyte spheroids. IL-1β-stimulated mouse articular chondrocytes were treated with different concentrations of liraglutide for 24h. Production of matrix metalloproteinase MMP-13, prostaglandin E2 (PGE2) and nitrite was measured by ELISA and Griess reaction, respectively. Exendin 9-39, a GLP-1R antagonist, was used to confirm target engagement in thein vitroexperiments. Intra-articular (IA) injections of liraglutide or vehicle were performed in the type II collagenase rat model. Histopathological analyses (OARSI scores1) were conducted blindly by one investigator.Results:Liraglutide induced the differentiation of hMSCs into chondrocytes. Indeed, 21 days after differentiation initiation, 5/6 and 4/6 alcian-blue positive spheroids were observed for 10 and 100nM liraglutide, respectively, versus 0/6 for vehicle. Liraglutide significantly reduced dose-dependently the IL-1β-induced production of PGE2 (5808±178 for vehicle vs 4560±140, 2933±171 and 2365±85 pg/ml for liraglutide 10, 100 and 500nM, respectively, p≤0.001), nitrite (24.9±0.4 for vehicle vs 20.9±1.5, 19.1±0.9 and 16.5±0.5 µM for liraglutide 10, 100 and 500nM, respectively, p≤0.001) and MMP-13 (686±9 for vehicle vs 553±3, 402±5 and 297±8 pg/ml for liraglutide 10, 100 and 500nM, respectively, p≤0.001) in murine chondrocytes. Effects of liraglutide were GLP-1R dependent since exendin 9-39 significantly counteracted both chondrogenesis and inflammation/catabolism markers expression. Histological assessment of rat collagenase-injected knee joint revealed a significant (p≤0.05) decrease of the total joint score in the IA Liraglutide treated group (8±4) compared to vehicle (11±4).Conclusion:Liraglutide induced chondrogenesis, decreased metalloproteinase and inflammatory mediators production by chondrocytes and protected cartilage inin vitroandin vivopreclinical OA models, opening the way for repositioning this drug as a potential DMOAD.References:[1]Osteoarthritis Cartilage. 2010 Oct;18 Suppl 3:S24-34Acknowledgments:All the people who contributed to the InOsteo project: the members of 4P-Pharma, INSERM UMR S938 research team, SATT Lutech and Sorbonne UniversityDisclosure of Interests:Francis Berenbaum Grant/research support from: TRB Chemedica (through institution), MSD (through institution), Pfizer (through institution), Consultant of: Novartis, MSD, Pfizer, Lilly, UCB, Abbvie, Roche, Servier, Sanofi-Aventis, Flexion Therapeutics, Expanscience, GSK, Biogen, Nordic, Sandoz, Regeneron, Gilead, Bone Therapeutics, Regulaxis, Peptinov, 4P Pharma, Paid instructor for: Sandoz, Speakers bureau: Novartis, MSD, Pfizer, Lilly, UCB, Abbvie, Roche, Servier, Sanofi-Aventis, Flexion Therapeutics, Expanscience, GSK, Biogen, Nordic, Sandoz, Regeneron, Gilead, Sandoz, Coralie Meurot Employee of: 4P-Pharma, Jerome Breton Employee of: 4P-Pharma, Laure Sudre: None declared, Carole Bougault: None declared, Revital Rattenbach Shareholder of: 4P-Pharma, Employee of: 4P-Pharma, Celine Martin Employee of: 4P-Pharma, Claire Jacques: None declared

scholarly journals LARYNGOTRACHEITIS VIRUS IN CHICKENS

1967 ◽  
Vol 125 (3) ◽  
pp. 409-428 ◽  
Author(s):  
Betsy G. Bang ◽  
Frederik B. Bang
Keyword(s):  
Alcian Blue ◽  
Plasma Cells ◽  
Giant Cells ◽  
Functional Restoration ◽  
Nasal Fossa ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Cell Nuclei ◽  
Histochemical Change ◽  
Laryngotracheitis Virus

Infectious laryngotracheitis can be produced in chickens as an experimental model of severe nonfatal rhinitis and sinusitis. Inoculated intranasally into unanesthetized baby chicks it remains limited to the nasal fossa, produces acute desquamation of all nasal epithelia, results in functional recovery of the respiratory epithelium, but leaves important residual abnormalities. From the earliest recognizable lesions through 4½ months' convalescence, the principal changes are as follows: 1. Initial lesions, or small syncytia of intranuclear "inclusions", first identifiable in the mucociliated cells of the shallowest portion of the epithelium at about 21 hr postinoculum (the inner surface of the maxillary conchal scroll). 2. Acute sloughing, (about 3 to 7 days), marked by: (a) spread of lesions from cell to cell via multinucleated "giant cells" which progressively slough and desquamate respiratory, olfactory, and sinus epithelia, epithelial neural elements and blood vessels; (b) appearance of numbers of eosinophilic leukocytes along the basement membrane at the sites of lesions just previous to sloughing; intensive infiltration of the submucosa with small lymphocytes after sloughing begins; (c) histochemical change in the intracellular mucus of the cells which comprise the syncytia: this mucus stains with Alcian blue alone when stained with AB-PAS; and (d) all cartilages of the maxillary conchae become flaccid, and the cell nuclei and matrix lose both basophilic and Alcian blue staining properties, effects which recede by about the 8th day. 3. Repair (about 8 to 21 days), marked by rapid initial spread of a sheet of epithelial cells over the infiltrated subrmucosa, appearance of numbers of plasma cells circulating in the tissues, formation of encapsulated secondary nodules, and mucosal adhesions. 4. Convalescence (about 1 to 4½ months when experiments terminated), marked by functional restoration of the mucociliary lining of the nasal fossa. However, at 4½ months eight specimens all show complete metaplasia of the olfactory organ (end nerves, supporting cells, and glands of Bowman) to mucociliated epithelium, all show abnormal formation and alignment of mucous acini, and about 50% have severe persistent sinusitis.

Histochemical observations on the tegumentary epithelium and interproglottidal glands of Moniezia expansa (Rud., 1805) (Cestoda, Cyclophyllidea)

Parasitology ◽  
1969 ◽  
Vol 59 (3) ◽  
pp. 505-518 ◽  
Author(s):  
R. E. Howells ◽  
D. A. Erasmus
Keyword(s):  
Alcian Blue ◽  
Regional Differences ◽  
Succinic Dehydrogenase ◽  
Neck Region ◽  
Secretory Activity ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Light Microscope Level ◽  
Gland Cells ◽  
Moniezia Expansa

Regional differences in the tegumentary tissue of Moniezia expansa, as revealed at the light-microscope level by histological and histochemical techniques, are described and evidence for secretory activity by the interproglottidal glands is presented.In very immature proglottides the interproglottidal glands are at the ‘precryptic’ stage. Gland cells may be differentiated from other tegumentary cells by their high RNA content and in certain gland cells the presence of an alcian blue staining material.In mature proglottides the glands consist of rosette-like clusters of cells around crypt-like intuckings of the tegument. Two types of cells are found in the gland, small alcian blue-staining cells which are most numerous in the neck region of the crypt, and larger cells, the predominant gland cells, which do not stain with alcian blue but possess non-specific esterase activity. No other tegumentary cells in Moniezia exhibit this activity. Esterase and phosphatase activity is found in the tegument and crypt of the glands and in the interproglottidal folds.The non-enzyme histochemistry confirms and extends the observations of previous workers.Cytochrome oxidase and succinic dehydrogenase were detected in the tegumentary cells and tegument. Very strong reactions were given in the neck and scolex, with a progressive diminution of activity posteriorly along the strobila. Very low activities were recorded in the tegument of the glands.

Dual action of sonic hedgehog on chondrocyte hypertrophy:retrovirus mediated ectopic sonic hedgehog expression in limb bud micromass culture induces novel cartilage nodules that are positive for alkaline phosphatase and type X collagen

1997 ◽  
Vol 110 (21) ◽  
pp. 2691-2701 ◽  
Author(s):  
N.S. Stott ◽  
C.M. Chuong
Keyword(s):  
Alkaline Phosphatase ◽  
Gene Family ◽  
Alcian Blue ◽  
Sonic Hedgehog ◽  
Ectopic Expression ◽  
Limb Bud ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Type X Collagen ◽  
Chondrocyte Maturation

Members of the vertebrate hedgehog gene family (HH) are involved in patterning and modulation of differentiation. Recently it has been shown that ectopic expression of HH gene family members in vivo blocks chondrocyte maturation through activation of a parathyroid hormone related peptide (PTHrP) dependent negative regulatory loop in the perichondrium. However, the direct effect of HH on chondrocyte maturation has not been tested. Here, we studied the effect of retroviral overexpression of the chicken sonic hedgehog gene (Shh) on the growth and maturation of limb bud cells in micromass cultures. Shh is neither expressed nor required for the initiation of cellular condensation in normal micromass cultures. With Shh over-expression, micromass cultures developed novel tightly whorled nodules in addition to the normal Alcian Blue positive cartilage nodules. We characterized the new nodules and showed that they are strongly positive for alkaline phosphatase, enriched in type X collagen and weakly positive for Alcian Blue staining. Shh overexpression also increased cell proliferation, but this cannot account for the formation of the new nodules. This current study shows that misexpression of Shh in in vitro chondrogenic cultures promotes characteristics of hypertrophic chondrocytes. Thus HH has two complementary functions; a direct positive effect on chondrocyte hypertrophy in the absence of PTHrP pathway, and an indirect negative feedback loop through PTHrP to prevent other less differentiated chondrocytes from becoming hypertrophic. These two complementary actions of HH coordinate the progression of cartilage maturation.

Thyroid Follicular Hyperplasia Associated With Massive Extracellular Mucin Deposition

2017 ◽  
Vol 25 (6) ◽  
pp. 533-535 ◽  
Author(s):  
Francesco Nesa ◽  
Luca Poggi ◽  
Stefano Ferrero ◽  
Alessandro Del Gobbo
Keyword(s):  
Alcian Blue ◽  
Periodic Acid ◽  
The Other ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Thyroid Tumors ◽  
Stromal Compartment ◽  
Follicular Hyperplasia ◽  
Nodular Hyperplasia ◽  
Thyroid Parenchyma

Extensive extracellular mucin deposition is a rare pathological thyroid condition with 6 cases described in literature so far. We report another case of a 67-year-old woman, discussing histopathological features, and review the literature. Our findings showed a diffuse mucin deposition in the stromal compartment of thyroid parenchyma. Histochemical stainings showed positivity for Alcian blue staining, but not for periodic acid–Shiff staining. Our case is peculiar because this mucin deposition was associated with benign nodular hyperplasia, in contrast with the other 6 reports, which described the same stromal alterations associated with benign or malignant thyroid tumors.

scholarly journals Η χρήση του πεπτιδίου Ec της ισόμορφης IGF-1Ec στην επισκευή χόνδρινων βλαβών

2018 ◽  
Author(s):  
Νικόλαος Αρμακόλας
Keyword(s):  
Polymerase Chain Reaction ◽  
Trypan Blue ◽  
Quantitative Polymerase Chain Reaction ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Tgf Β1 ◽  
Invasion Assays

Το πεπτίδιο Ec (PEc) του IGF-1Ec (IGF-1Ec) επάγει την κινητοποίηση των ανθρωπίνων μεσεγχυματικών βλαστικών κυττάρων (hMSC) και ενεργοποιεί την εξωκυτταρική κινάση 1 και 2 (ERK 1/2) διαφόρων κυττάρων. Σκοπός της παρούσας μελέτης ήταν η διερεύνηση της επιδρασης του PEc στην κινητοποίηση και τη διαφοροποίηση των hMSCs, καθώς και η δυνατότητα εφαρμογής του σε συνδυασμό με τον TGF-β1 (TGF-β1) στην επιδιόρθωση του αρθρικού χόνδρου. Τα αποτελέσματα της εξωγενούς χορήγησης του ΡΕc και του ΤGF-β1, ξεχωριστά και σε συνδυασμό, σε hMSCs εκτιμήθηκαν χρησιμοποιώντας trypan blue assay, reverse transcription-quantitative polymerase chain reaction, western blot analysis, Alcian blue staining, wound healing assays και migration/invasion assays. Προσδιορίστηκε ότι το PEc εμπλέκεται στη διαδικασία διαφοροποίησης των hMSCs προς υαλώδη χόνδρο. Η χορήγηση PEc ή / και TGF-β1 σε hMSCs έδειξε συγκρίσιμη εναπόθεση χονδρικής θεμέλειας ουσίας. Ακόμα, η χορήγηση του ΡΕc σε συνδυασμό με τον ΤGF-β1 συσχετίστηκε με μια σημαντική αύξηση στην κινητοποίηση των hMSC σε σύγκριση με την χορήγηση μόνο του TGF-β1 ή του ΡEc (Ρ <0,05). Επομένως, το ΡΕc φαίνεται να διευκολύνει in vitro την κινητοποίηση των hMSC και την διαφοροποίηση τους προς χονδροκύτταρα, ενισχύοντας το ρόλο του ΤGF-β1.

scholarly journals Alizarin Red S-Alcian Blue Staining for Regenerated tail of Common House Gecko (Hemidactylus frenatus)

2018 ◽  
Vol 7 (2) ◽  
pp. 57-59
Author(s):  
Rakhmiyati Rakhmiyati ◽  
Muhammad Ja’far Luthfi
Keyword(s):  
Alcian Blue ◽  
Histochemical Staining ◽  
Alizarin Red S ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Tail Regeneration ◽  
Alizarin Red ◽  
Regenerate Tail ◽  
Hemidactylus Frenatus ◽  
Common House

Common House Gecko (Hemidactylus frenatus) is one of reptiles that have ability to autotomy their tails. Tail autotomy is a mechanism to protect it self from predators. After the tail broke, there will be wound healing on the tail which is then followed by a tail regeneration event. Original tail and regenerate tail is very different morphologically and anatomically. The original tail is composed of bones while the tail of the regenerate is composed of cartilage. Histochemical staining using Alizarin Red-S Alcian Blue was done to differentiate bone and cartilage. This method will stained bones red while the cartilage will stained blue.

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