AIMTo analyse the cytomorphological features of keratinocytes in smears obtained from the oral mucosa of tobacco users and from oral squamous cell carcinoma lesions.

METHODOLOGYOral smears were obtained from clinically, normal appearing mucosa of oral squamous cell carcinoma patients (n=20) and from the mucosa of smokers (n=20), and apparently healthy individuals (n=20) were used as controls. The smears were histochemically stained and cytomorphological assessment of the keratinocytes was carried out. One-way ANOVA (Analysis of Variance) was used for comparing the parameters among multiple groups and Tukey-HSD test was used to compare the mean values between groups.

RESULTSThe mean nuclear area of keratinocytes from the mucosa of tobacco users was 46 +/- 2.57 and that of the oral squamous cell carcinoma lesion was 81.54 +/- 4.31. While there was a significant (P = 0.001) reduction in the cellular area of keratinocytes from oral squamous cell carcinoma lesion when compared with those from oral smears of tobacco users.

CONCLUSIONCytomorphometric analysis of keratinocytes can serve as a useful adjunct in the early diagnosis of oral squamous cell carcinomas.

Azo Compounds ; Carcinoma, Squamous Cell ; pathology ; Cell Nucleus ; ultrastructure ; Cell Size ; Coloring Agents ; Cytodiagnosis ; instrumentation ; methods ; Cytoplasm ; ultrastructure ; Eosine Yellowish-(YS) ; Fluorescent Dyes ; Humans ; Image Processing, Computer-Assisted ; methods ; Keratinocytes ; pathology ; Mouth Mucosa ; pathology ; Mouth Neoplasms ; pathology ; Rosaniline Dyes ; Smoking ; pathology ; Software ; Tobacco, Smokeless

INTRODUCTIONAn investigation was carried out to determine the morphological characteristics of fibroblasts in two portions of the vocal fold (VF) mucosa, the macula flava (MF) and Reinke's space (RS), of three different age groups: newborns, adults and geriatrics.

METHODSNormal human VF obtained from autopsy cases were included in this study: four from mature newborns; four from middle-aged adults; and four from geriatric cases. Fibroblasts in RS and MF were investigated by transmission electron microscopy.

RESULTSThe fibroblasts of the MF in both adults and newborns tended to be stellate in shape, with a small nucleus/cytoplasm (N/C) ratio and a well-developed rough endoplasmic reticulum (rER) and Golgi apparatus (GA). Most of the fibroblasts present in RS were oval in newborns and spindle-shaped in adults, with a large N/C ratio and less developed rER and GA. The majority of fibroblasts of the geriatric MF were stellate in shape; while in geriatric RS, the majority of fibroblasts were spindle-shaped with an N/C ratio of 0.5 to 2.0 as in the case of adults. However, the development of rER and GA was less marked in geriatrics than in adults.

CONCLUSIONHistological changes of fibroblasts in the VF mucosa are one of the important causes of the change in voice quality with ageing. Furthermore, geriatric changes in the vocal ligament can be attributed to the activities and the presence of ageing processes in fibroblasts of geriatric VF mucosa.

Adult ; Age Factors ; Aged ; Cell Nucleus ; ultrastructure ; Cytoplasm ; ultrastructure ; Endoplasmic Reticulum ; ultrastructure ; Female ; Fibroblasts ; ultrastructure ; Golgi Apparatus ; ultrastructure ; Humans ; Infant, Newborn ; Laryngeal Mucosa ; ultrastructure ; Male ; Microscopy, Electron, Transmission ; methods ; Vocal Cords ; ultrastructure

AIMTo detect the expression of HSP25 in rat dental follicles both in vivo and vitro, and explore the underlying mechanism of HSP25 on the proliferation and differentiation of rat dental follicle cells (DFCs).

METHODOLOGYImmunohistochemistry was performed to detect the expression of HSP25 in mandibles of postnatal rats on days 1, 3, 5, 7, 9 and 11 in vivo. In vitro, the expression of HSP25 in DFCs was detected by an indirect immunofluorescence assay. Thiazolyl blue tetrazolium bromide (MTT) assay, flow cytometry and alkaline phosphatase (ALP) assay were used to identify the time-course effect mediated by different concentrations of recombinant murine HSP25 of 0, 1, 10, 50 and 100 ng/mL on rat DFCs.

RESULTSExpression of HSP25 was not detected in dental follicles of the rats until day 5 after birth, but became up-regulated in a time-dependent manner till day 11. HSP25 was detected in the cytoplasm of cultured rat DFCs. No significant difference could be observed in the proliferation of DFCs after stimulation with different concentrations of HSP25 on days 1, 2 and 3 (P > 0.05). HSP25 at concentrations of 50 ng/mL and 100 ng/mL up-regulated the ALP activity of DFCs on day 9 (P < 0.05).

CONCLUSIONHSP25-immunoreactivity increased chronologically during the development of dental follicles. The protein had no significant effect on cell proliferation but may play a role in cementoblast/osteoblast differentiation of DFCs.

Alkaline Phosphatase ; analysis ; Ameloblasts ; cytology ; Animals ; Cell Culture Techniques ; Cell Differentiation ; physiology ; Cell Proliferation ; Coloring Agents ; Cytoplasm ; ultrastructure ; Dental Sac ; cytology ; growth & development ; Flow Cytometry ; Fluorescent Antibody Technique, Indirect ; HSP27 Heat-Shock Proteins ; analysis ; physiology ; Odontoblasts ; cytology ; Rats ; Rats, Sprague-Dawley ; Tetrazolium Salts ; Thiazoles ; Tooth Germ ; cytology ; Up-Regulation ; physiology

OBJECTIVEIt is well known that glioma stem cells-progenitors (GSCP) proliferate indefinitely and hardly differentiate in vitro, however, the reasons remain unknown. The aim of this study was to explore the ultrastructural basis of GSCP.

METHODSGSCP, kept by our laboratory, were collected, embedded, and cut into ultrathin sections and observed under the transmission electron microscope.

RESULTSA single GSCP usually had relatively well developed mitochondria, Golgi apparatuses, ribosomes, and undeveloped rough endoplasmic reticulum, but seldom lysosomes and no typical autophagosomes were found, and the nuclear-cytoplasmic ratio was high. The nuclei frequently contained huge amounts of euchromatin and a small quantity of heterochromatin, and in most nuclei there were only one nucleolus, however, two or more nucleoli were also common. Typical apoptotic cells could hardly be found in tumor-spheres, and between neighboring cells in tumor-spheres there were incompletely developed desmosomes or intermediate junction.

CONCLUSIONThe ultrastructural features of glioma stem cells-progenitors showed that BTSCP were very primitive and the lack of autophagy and the underdevelopment of some other cellular organelles are probably the reasons for the differential inhibition of GSCPs.

Brain Neoplasms ; ultrastructure ; Cell Line, Tumor ; Cell Membrane ; ultrastructure ; Cell Nucleus ; ultrastructure ; Chromatin ; ultrastructure ; Cytoplasm ; ultrastructure ; Glioma ; ultrastructure ; Humans ; Intercellular Junctions ; ultrastructure ; Microscopy, Electron, Transmission ; Mitochondria ; ultrastructure ; Neoplastic Stem Cells ; ultrastructure

A marine unicellular green alga, Platymonas subcordiformis, was demonstrated to photobiologically produce hydrogen gas from seawater. The objective of this study was to localize and identify the hydrogenase isolated from P. subcordiformis. Adaptation in the presence of inhibitors of protein biosynthesis indicated that the hydrogenase was much more inhibited by cycloheximide than that by chloramphenicol. The result suggested that the hydrogenase isolated from P. subcordiformis is probably synthesized in cytoplasmic ribosomes. Both Western blot analysis and immunogold electron microscopy demonstrate that the P. subcordiformis hydrogenase is mainly located in the chloroplast stroma. The proteins that reacted specifically with the antibodies against the iron hydrogenase isolated from Chlamydomonas reinhardtii were concentrated by immunoprecipitation. The separated protein bands were cut out of the SDS-PAGE gel, in-gel digested by trypsin, and analyzed by Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS). Mascot was employed for analysis of the MALDI data using the public databases NCBInr. The hydrogenase isolated from P. subcordiformis was identified to be the Fe-hydrogenase.
Algal Proteins ; isolation & purification ; metabolism ; Biocatalysis ; drug effects ; Blotting, Western ; Chloramphenicol ; pharmacology ; Chlorophyta ; enzymology ; Cycloheximide ; pharmacology ; Cytoplasm ; enzymology ; ultrastructure ; Electrophoresis, Polyacrylamide Gel ; Hydrogenase ; antagonists & inhibitors ; isolation & purification ; metabolism ; Immunoprecipitation ; methods ; Iron-Sulfur Proteins ; antagonists & inhibitors ; isolation & purification ; metabolism ; Kinetics ; Microscopy, Immunoelectron ; Protein Synthesis Inhibitors ; pharmacology ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods

Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors in the gastrointestinal tract (GIT). Although interstitial cells of Cajal has been suggested as origin of this tumor, the cytological and ultrastructural features of GISTs are heterogeneous and unclear. A total 10 cases of normal gastrointestinal tissue (control), 13 GISTs of the stomach (8), small intestine (3), mesocolon (1) and liver (1), and 2 gastrointestinal autonomic nervous tumor (GANT) of small intestine were ultrastructurally studied. Normal interstitial cells of Cajal (ICC) were abundantly present around the myenteric plexuses or individually scattered through the wall of GIT. ICC was characterized by slender cytoplasmic processes, well-developed endoplasmic reticulum (ER), mitochondria, Golgi apparatus, caveolae and intermediate filaments. The GISTs and GANTs had overlapping ultrastructures. The most common and important ultrastructural features of GISTs were rich villous cytoplasmic processes, dispersed intermediate filaments and abundant SER, and those of GANTs were neurosecretory granules and skenoid fibers. Compared with ICC, the GISTs and GANTs had remarkably reduced caveolae and gap junctions. Our study suggested that ultrastructural analysis gives much information to investigate lineage differentiation of neoplastic cells and make a differential diagnosis of these tumors from other mesenchymal tumors and between GISTs and GANTs.
Adult ; Aged ; Autonomic Nervous System/*pathology/ultrastructure ; Comparative Study ; Cytoplasm/pathology/ultrastructure ; Female ; Gastrointestinal Neoplasms/*pathology/ultrastructure ; Human ; Immunohistochemistry ; Male ; Microscopy, Electron ; Middle Aged ; Peripheral Nervous System Neoplasms/*pathology/ultrastructure ; Stromal Cells/*pathology/ultrastructure ; Support, Non-U.S. Gov't ; Tumor Markers, Biological ; Vacuoles/pathology/ultrastructure

To identify the knowledge of rare lymphoproliferative disorder, the clinical and biological features of three kinds of lymphoproliferative disorders with cytoplasmic projections were compared. The clinical manifestations, ultrastructure and immunophenotype were analyzed. The results showed that hairy cell leukemia (HCL), splenic lymphoma with villous lymphocyte (SLVL) and hairy cell leukemia-variant (HCL-V) had some common characters including splenomegaly, peripheral blood and bone marrow infiltration by villous lymphocyte and B lymphocyte immunophenotype; but these three disorders had specific features respectively. It was concluded that overall analysis of clinical and laboratory features might be contributive to the differential diagnosis of these three disorders.
Adolescent ; Adult ; Aged ; Antibodies, Neoplasm ; blood ; Antigens, Differentiation, B-Lymphocyte ; blood ; Antigens, Neoplasm ; blood ; B-Lymphocytes ; immunology ; ultrastructure ; Bone Marrow Cells ; pathology ; Cytoplasm ; ultrastructure ; Female ; Humans ; Immunophenotyping ; Leukemia, Hairy Cell ; immunology ; pathology ; Lymphoma, B-Cell ; immunology ; ultrastructure ; Lymphoproliferative Disorders ; immunology ; pathology ; Male ; Middle Aged

A hybrid segmentation algorithm is proposed for automatic segmentation of blood cell images based on adaptive multi-scale thresholding and seeded region growing techniques. Firstly, an adaptive and scale space filter (ASSF) is applied to image histogram and a scale space image is built. According to the properties of the scale space image, proper thresholds can be obtained to separate the nucleus from the original image and the white blood cells are located. Secondly, the local color similarity and global morphological criteria constrain seeded region growing in order to finish the segmentation of the cytoplasm. The detection accuracy of white blood cell is 98% and the segmentation accuracy based on the subjective evaluation is 93%. Test shows that this algorithm is effective for automatic segmentation of white blood cells.
Algorithms ; Automation ; Blood Cells ; Cell Nucleus ; ultrastructure ; Color ; Cytoplasm ; ultrastructure ; Humans ; Image Enhancement ; Leukocytes

Histochemical, immunohistochemical and ultrastructural studies were performed on cases of hepatocellular carcinoma (HCC) with pale bodies (PB). HCC containing PBs was observed in 3 (5.5%) of 55 consecutively resected HCC cases. Histologically, a large number of hepatocytes displayed pale or eosinophilic staining of the cytoplasm, resulting in ground-glass appearance. They were aggregated in nodular pattern, or diffusely intermixed with other malignant hepatocytes. PBs were negative for periodic-acid Schiff and Masson's trichrome staining. The inclusions showed a strong positive reaction for fibrinogen and some of them were weakly positive for albumin but negative for hepatitis B surface antigen, hepatitis B core antigen, alpha-fetoprotein and alpha-1-antitrypsin. Ultrastructurally, PBs were membrane-bound and contained granular materials of moderate electron density, and were closely related to dilated rough endoplasmic reticulum. These findings support that PBs are secretory fibrinogen accumulated in cystic ER and that such intracellular accumulation possibly reflects a defective transport of fibrinogen.
Albumins/analysis ; Carcinoma, Hepatocellular/pathology* ; Cytoplasm/ultrastructure ; Cytoplasm/pathology ; Cytoplasm/chemistry ; Endoplasmic Reticulum, Rough/ultrastructure ; Endoplasmic Reticulum, Rough/pathology ; Endoplasmic Reticulum, Rough/chemistry ; Fibrinogen/analysis ; Human ; Inclusion Bodies/ultrastructure ; Inclusion Bodies/pathology* ; Inclusion Bodies/chemistry ; Liver Neoplasms/pathology* ; Male ; Microscopy, Electron ; Middle Age ; Periodic Acid-Schiff Reaction

Animals ; Cytoplasm ; ultrastructure ; Fibroblasts ; ultrastructure ; Fractures, Bone ; pathology ; Microscopy, Electron ; Osteoblasts ; ultrastructure ; Rabbits ; Wound Healing