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The Journal of Experimental Medicine Jun 1962Lymph nodes and splenic tissue from patients with congenital agammaglobulinemia and dysgammaglobulinemia and from normal subjects were studied with the use of...
Lymph nodes and splenic tissue from patients with congenital agammaglobulinemia and dysgammaglobulinemia and from normal subjects were studied with the use of immunofluorescence and histochemical stains to determine the site of synthesis of the 19S gamma(1)-globulins. The two patients with dysgammaglobulinemia had high serum concentrations of the 19S gamma(1)-globulins and a marked deficit of the 7S gamma-globulins. These patients, as well as agammaglobulinemic children, had only rare or no plasma cells in their tissues. Cells were identified in sections of spleen from a dysgammaglobulinemic child as well as from normal individuals which exhibited specific fluorescence with an anti-19S gamma-globulin antiserum adsorbed with 7S gamma(2)-globulins and which stained positively with PAS and methyl green pyronine. These cells resembled the transitional cells described by Fagraeus.
Topics: Agammaglobulinemia; Dysgammaglobulinemia; Fluorescent Antibody Technique; Genetic Diseases, X-Linked; Humans; Immune Sera; Immunoglobulin G; Immunoglobulin M; Lymph Nodes; Plasma Cells; Spleen; gamma-Globulins
PubMed: 13882451
DOI: 10.1084/jem.115.6.1141 -
Medicina Oral, Patologia Oral Y Cirugia... Nov 2008Keratocystic odontogenic tumors (KOTs), also known as odontogenic keratocysts, were recently classified as a benign neoplasia due to the aggressive clinical behavior....
OBJECTIVES
Keratocystic odontogenic tumors (KOTs), also known as odontogenic keratocysts, were recently classified as a benign neoplasia due to the aggressive clinical behavior. Although several studies have shown the high proliferative activity of the epithelial lining, few studies have evaluated apoptosis in KOTs. Therefore, the aim of this study is to evaluate and compare the proliferation index (PI) and the apoptotic index (AI) of the epithelial lining in sporadic KOTs, KOTs associated with the Nevoid Basal Cell Carcinoma Syndrome (NBCCS KOTs), and dentigerous cysts.
MATERIAL AND METHODS
A total of 11 sporadic KOTs, 15 NBCCS KOTs, and 11 dentigerous cysts were evaluated. The PI was assessed by immunohistochemical detection of the cell proliferation marker Ki-67. The AI was assessed by morphological evaluation of sections stained by methyl green-pyronin. The TUNEL assay was used to confirm the occurrence of apoptosis. Differences in the PI and the AI between sporadic KOTs, NBCCS KOTs, and dentigerous cysts were analyzed using the Kruskal-Wallis test. Differences in the PI and the AI between the epithelial layers of each lesion were analyzed using the Wilcoxon test.
RESULTS
The PI and AI were higher in sporadic and NBCCS KOTs than in dentigerous cysts. No difference in these indexes was observed between sporadic and NBCCS KOTs. In dentigerous cysts, the PI was higher in the basal layer. In sporadic and NBCCS KOTs, the PI was higher in suprabasal layer. No difference in the AI was observed between the basal layer and the suprabasal layer in the three lesions. The AI was higher in the superficial layer of sporadic and NBCCS KOTs.
CONCLUSIONS
The present study demonstrates that the epithelial lining of KOTs shows a distinct pattern of cell proliferation and apoptosis, reflecting its high cell turnover and reinforcing its classification as an odontogenic tumor.
Topics: Apoptosis; Basal Cell Nevus Syndrome; Cell Proliferation; Humans; Mouth Neoplasms; Odontogenic Tumors
PubMed: 18978709
DOI: No ID Found -
Journal of Bone and Joint Infection 2018Visualization of biofilm using histochemical staining and combined histochemistry (HC) and immunohistochemistry (IHC). The ability of S54F9 to form biofilm was...
Visualization of biofilm using histochemical staining and combined histochemistry (HC) and immunohistochemistry (IHC). The ability of S54F9 to form biofilm was tested . Hereafter, infected bone tissue was collected from two different porcine models of osteomyelitis inoculated with strain S54F9. The infection time was five and fifteen days, respectively. Twenty-five different histochemical staining protocols were tested in order to find the stains that could identify extracellular biofilm matrix. Protocols with an optimal visualization of biofilm extracellular matrix were combined with an immunohistochemical protocol based on a specific antibody against The combined protocols were applied to the tissue from the porcine models and to infected bone tissue from a child suffering from chronic staphylococcal osteomyelitis for more than a year. S54F9 showed an ability to form biofilm . Visualization of biofilm, . bacterial cells and extracellular matrix in different colours, was seen when the immunohistochemical protocol was combined with Alcian Blue pH3, Luna and Methyl-pyronin green. The bacterial cells were red to light brown and the extracellular matrix either light blue, blue or orange depending on the histochemical stain. In the porcine models and the human case 10 and 90 percent, respectively, of the bacterial aggregates in a 100x magnification field displayed both the extracellular matrix and the bacterial cells simultaneously in two different colours. A combination of HC and IHC can be used to diagnose and characterise biofilm infections on a routine basis.
PubMed: 29545993
DOI: 10.7150/jbji.22799 -
Journal of Clinical Microbiology Feb 1979A total of 78 strains of Haemophilus vaginalis were examined for 104 features. All strains fermented dextrin, maltose, and starch. Additionally, more than 90% of the...
A total of 78 strains of Haemophilus vaginalis were examined for 104 features. All strains fermented dextrin, maltose, and starch. Additionally, more than 90% of the strains fermented galactose, glucose, and ribose. Arbutin, cellobiose, melibiose, rhamnose, and salicin were not fermented by any of these strains. None of the strains acidified any of 14 alcohols or alkalinized any of 25 organic salts and amides. More than 90% of the strains hemolyzed human blood agar and hydrolyzed hippurate. No strain hemolyzed sheep blood agar. A recommendation is included for those minimal features that best differentiate H. vaginalis from other oxidase- and catalase-negative, gram-negative organisms.
Topics: Bacitracin; Carbohydrate Metabolism; Coloring Agents; Drug Resistance, Microbial; Fermentation; Gardnerella vaginalis; Haemophilus; Haemophilus Infections; Haemophilus influenzae; Hemolysis; Humans; Pyronine
PubMed: 311779
DOI: 10.1128/jcm.9.2.200-204.1979 -
The American Journal of Pathology Dec 1991Numerous hepatic cell lineage pathways have been proposed for the development of hepatocarcinogensis induced by chemical carcinogens in rats. The roles of bile ductule...
Numerous hepatic cell lineage pathways have been proposed for the development of hepatocarcinogensis induced by chemical carcinogens in rats. The roles of bile ductule cells and hepatocytes in the development of carcinogenesis were investigated using light and electron microscopic procedures to detect differences in morphology and in the phenotypic expression of antigens that are associated with each cell type. In early stages of hepatocarcinogenesis (4-10 weeks after initiation of feeding of a choline-deficient ethionine containing diet), both bile ductulelike (BDL) cells and hepatocytes were seen in mitosis. At the light microscope level, BDL cells showed intense cytoplasmic pyronin (RNA) staining and were positive for the antigens defined by monoclonal antibody 270.38 (bile ductule cells and "oval" cell marker) and glutathione-S-transferase (Yp isoform), whereas hepatocytes were positive for the antigens defined by monoclonal antibodies 270.26 and 258.26 (liver parenchymal cell markers), catalase activity (peroxisome marker) and adenosine triphospatase activity (bile canalicular marker). The authors frequently encountered BDL cells and hepatocytes in close proximity. Ultrastructural examination showed extensive plasma membrane appositions between a subset of BDL cells and hepatocytes. Desmosome structures, tight junctions, microvilli interdigitations and ATPase-positive bile canalicularlike structures were present along the contiguous plasma membrane domains of BDL cells and hepatocytes. Many of the BDL cells attached to hepatocytes were also attached to other BDL cells that had retained a basal lamina. In many cases, BDL cells connected to both hepatocytes and other BDL cells were no longer completely surrounded by basal lamina and had acquired a dual polarity as a consequence of their sharing apical and lateral membrane domains with both BDL cells and hepatocytes. BDL cells showed increased numbers of microperoxisomes (catalase positive organelles) and numerous free ribosomes. Hepatocytes showed a prominent development of the smooth endoplasmic reticulum, a feature prominent in hepatocytes within hyperplastic nodules. Since BDL cells and hepatocytes proliferate and BDL cells and hepatocytes develop intercellular junction sites, the authors propose that both cell types in early stages of carcinogenesis have the capacity to enter the cell lineage pathway leading to the development of hepatocarcinoma. Furthermore, the finding that BDL cells and hepatocytes form multiple attachment sites at the level of the plasma membrane, suggests the possibility that at some stage convergence of separate hepatic cell pathways may occur.
Topics: Animals; Antigens; Bile Ducts; Carcinoma; Cell Communication; Cell Membrane; Cell Polarity; DNA; Ethionine; Lipids; Liver; Liver Neoplasms; Microbodies; Microscopy, Electron; Rats; Ribosomes
PubMed: 1750508
DOI: No ID Found -
Acta Biologica Hungarica Mar 2016This study describes the macroscopic anatomy and the microscopic and ultrastructural features of the Harderian gland and lacrimal gland of the Capercaillies. It was...
This study describes the macroscopic anatomy and the microscopic and ultrastructural features of the Harderian gland and lacrimal gland of the Capercaillies. It was conducted both on adult male and female Capercaillies. Tissue sections were stained with hematoxylin and eosin, azan trichrome, modified Mallory's trichrome, methyl green-pyronin Y, periodic acid-Schiff, alcian blue pH 2.5, aldehyde fuchsin and Hale's dialysed iron. The morphometric study of the Harderian and lacrimal glands indicated that they are both larger in male than in female Capercaillies. The histological analysis showed that the HG has a multilobar tubulo-alveolar structure with numerous lymphocytes and plasma cells. The LG has a multilobar tubulo-acinar structure without lymphocytes and plasma cells. The periodic acid-Schiff staining and alcian blue pH 2.5 staining demonstrated a mild positive reaction in the epithelial cells of the Harderian gland and weak positive reaction in the lacrimal gland. The HDI staining detected the presence of carboxylated acid mucopolysaccharides in the Harderian and lacrimal glands. Transmission electron microscopy revealed the presence of two types of secretory vesicles in the cytoplasm of both studied glands. It also showed that lipid droplets and glycogen granules were more abundant in the Harderian gland than in the lacrimal gland of this species.
Topics: Animals; Female; Galliformes; Harderian Gland; Lacrimal Apparatus; Male; Microscopy, Electron, Transmission
PubMed: 26960354
DOI: 10.1556/018.67.2016.1.2 -
Bioorganic & Medicinal Chemistry Letters Jul 20249-cyanopyronin is a promising scaffold that exploits resonance Raman enhancement to enable sensitive, highly multiplexed biological imaging. Here, we developed...
9-cyanopyronin is a promising scaffold that exploits resonance Raman enhancement to enable sensitive, highly multiplexed biological imaging. Here, we developed cyano-Hydrol Green (CN-HG) derivatives as resonance Raman scaffolds to expand the color palette of 9-cyanopyronins. CN-HG derivatives exhibit sufficiently long wavelength absorption to produce strong resonance Raman enhancement for near-infrared (NIR) excitation, and their nitrile peaks are shifted to a lower frequency than those of 9-cyanopyronins. The fluorescence of CN-HG derivatives is strongly quenched due to the lack of the 10th atom, unlike pyronin derivatives, and this enabled us to detect spontaneous Raman spectra with high signal-to-noise ratios. CN-HG derivatives are powerful candidates for high performance vibrational imaging.
Topics: Spectrum Analysis, Raman; Molecular Structure; Vibration; Nitriles
PubMed: 38636718
DOI: 10.1016/j.bmcl.2024.129757 -
Transactions of the American Clinical... 2012The hierarchical models of stem cell biology have been based on work first demonstrating pluripotental spleen-colony-forming units, then showing progenitors with many...
The hierarchical models of stem cell biology have been based on work first demonstrating pluripotental spleen-colony-forming units, then showing progenitors with many differentiation fates assayed in in vitro culture; there followed the definition and separation of "stem cells" using monoclonal antibodies to surface epitopes and fluorescent-activated cell characterization and sorting (FACS). These studies led to an elegant model of stem cell biology in which primitive dormant G0 stem cells with tremendous proliferative and differentiative potential gave rise to progressively more restricted and differentiated classes of stem/progenitor cells, and finally differentiated marrow hematopoietic cells. The holy grail of hematopoietic stem cell biology became the purification of the stem cell and the clonal definition of this cell. Most recently, the long-term repopulating hematopoietic stem cell (LT-HSC) has been believed to be a lineage negative sca-1+C-kit+ Flk3- and CD150+ cell. However, a series of studies over the past 10 years has indicated that murine marrow stem cells continuously change phenotype with cell cycle passage. We present here studies using tritiated thymidine suicide and pyronin-Hoechst FACS separations indicating that the murine hematopoietic stem cell is a cycling cell. This would indicate that the hematopoietic stem cell must be continuously changing in phenotype and, thus, could not be purified. The extant data indicate that murine marrow stem cells are continually transiting cell cycle and that the purification has discarded these cycling cells. Further in vivo BrdU studies indicate that the "quiescent" LT-HSC in G0 rapidly transits cycle. Further complexity of the marrow stem cell system is indicated by studies on cell-derived microvesicles showing that they enter marrow cells and transcriptionally alter their cell fate and phenotype. Thus, the stem cell model is a model of continuing changing potential tied to cell cycle and microvesicle exposure. The challenge of the future is to define the stem cell population, not purify the stem cell. We are at the beginning of elucidation of quantum stemomics.
Topics: Animals; Bone Marrow Cells; Cell Cycle; Cell Differentiation; Cell Proliferation; Cytoplasmic Vesicles; Hematopoietic Stem Cells; Humans; In Vitro Techniques; Mice; Phenotype; Stem Cells
PubMed: 23303982
DOI: No ID Found -
Pharmaceutics Nov 2021Pharmaceutical technology offers various dosage forms that can be applied interdisciplinary. One of them are spherical pellets which could be utilized as a carrier in...
Utilization of Pharmaceutical Technology Methods for the Development of Innovative Porous Metasilicate Pellets with a Very High Specific Surface Area for Chemical Warfare Agents Detection.
Pharmaceutical technology offers various dosage forms that can be applied interdisciplinary. One of them are spherical pellets which could be utilized as a carrier in emerging second-generation detection tubes. This detection system requires carriers with high specific surface area (SSA), which should allow better adsorption of toxic substances and detection reagents. In this study, a magnesium aluminometasilicate with high SSA was utilized along with various concentrations of volatile substances (menthol, camphor and ammonium bicarbonate) to increase further the carrier SSA after their sublimation. The samples were evaluated in terms of physicochemical parameters, their morphology was assessed by scanning electron microscopy, and the Brunauer-Emmett-Teller (BET) method was utilized to measure SSA. The samples were then impregnated with a detection reagent o-phenylenediamine-pyronine and tested with diphosgene. Only samples prepared using menthol or camphor were found to show red fluorescence under the UV light in addition to the eye-visible red-violet color. This allowed the detection of diphosgene/phosgene at a concentration of only 0.1 mg/m in the air for samples M20.0 and C20.0 with their SSA higher than 115 m/g, thus exceeding the sensitivity of the first-generation DT-12 detection tube.
PubMed: 34834274
DOI: 10.3390/pharmaceutics13111860 -
PloS One 2012Hematopoietic stem cells (HSCs) are responsible for sustaining hematopoietic homeostasis and regeneration after injury for the entire lifespan of an organism....
Hematopoietic stem cells (HSCs) are responsible for sustaining hematopoietic homeostasis and regeneration after injury for the entire lifespan of an organism. Maintenance of genomic stability is crucial for the preservation of HSCs, which depends on their efficient repair of DNA damage, particularly DNA double strand breaks (DSBs). Because of the paucity of HSCs and lack of sensitive assays, directly measuring the ability of HSCs to repair DSBs has been difficult. Therefore, we developed a sensitive and quantitative cell free in vitro non-homologous end joining (NHEJ) assay using linearized plasmids as the substrates and quantitative polymerase chain reaction (qPCR) technique. This assay can sensitively detect DSB repair via NHEJ in less than 1 µg 293T cell nuclear proteins or nuclear extracts from about 5,000 to 10,000 human BM CD34(+) hematopoietic cells. Using this assay, we confirmed that human bone marrow HSCs (CD34(+)CD38(-) cells) are less proficient in the repair of DSBs by NHEJ than HPCs (CD34(+)CD38(+) cells). In contrast, mouse quiescent HSCs (Pyronin-Y(low) LKS(+) cells) and cycling HSCs (Pyronin-Y(hi) LKS(+) cells) repaired the damage more efficiently than HPCs (LKS(-) cells). The difference in the abilities of human and mouse HSCs and HPCs to repair DSBs through NHEJ is likely attributed to their differential expression of key NHEJ DNA damage repair genes such as LIG4. These findings suggest that the qPCR-based cell free in vitro NHEJ assay can be used to sensitively measure the ability of human and mouse HSCs to repair DSBs.
Topics: Animals; Biological Assay; Blotting, Western; Bone Marrow; Cells, Cultured; DNA Breaks, Double-Stranded; DNA End-Joining Repair; DNA Helicases; DNA-Activated Protein Kinase; Flow Cytometry; Hematopoietic Stem Cells; Humans; In Vitro Techniques; Male; Mice; Mice, Inbred C57BL; Nuclear Proteins; RNA, Messenger; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction
PubMed: 22448248
DOI: 10.1371/journal.pone.0033499