Fine needle aspiration cytologic diagnosis of ganglioneuroblastoma

Two children with ganglioneuroblastomas in different locations underwent fine needle aspiration (FNA). Clinically, the tumor in the presacral area was diagnosed as a benign teratoma and the retroperitoneal tumor as a neuroblastoma. Both tumors were correctly diagnosed preoperatively as ganglioneuroblastomas by FNA cytology. The smears showed the characteristic Homer-Wright rosettes, ganglion cells and fibrillar material.     

Multiphoton tomography visualizes collagen fibers in the tumor microenvironment that maintain cancer‐cell anchorage and shape

Second harmonic generation (SHG) multiphoton imaging can visualize fibrillar collagen in tissues. SHG has previously shown that fibrillar collagen is altered in various types of cancer. In the present study, in vivo high resolution SHG multi‐photon tomography in living mice was used to study the relationship between cancer cells and intratumor collagen fibrils. Using green fluorescent protein (GFP) to visualize cancer cells and SHG to image collagen, we demonstrated that collagen fibrils provide a scaffold for cancer cells to align themselves and acquire optimal shape. These results suggest a new paradigm for a stromal element of tumors: their role in maintaining anchorage and shape of cancer cells that may enable them to proliferate. J. Cell. Biochem. 114: 99–102, 2012. © 2012 Wiley Periodicals, Inc.     

Myoepithelioma of the parotid gland initially diagnosed by fine needle aspiration cytology and immunocytochemistry: a case report

BACKGROUND: Myoepithelioma is a rare, benign tumor of the salivary gland, most commonly affecting the parotid gland. Although the cytologic features of myoepithelioma are documented in a few case reports, it has rarely been diagnosed preoperatively by fine needle aspiration (FNA) cytology. CASE: A 33-year-old man presented with a left parotid swelling 2.5 cm in diameter and of about 5 years' duration. FNA smears showed bundles of spindle-shaped cells as well as plasmacytoid and stellate cells in sheets and dissociated forms. A few cells had nuclear grooves, and occasional cells showed intranuclear cytoplasmic inclusions. In May-Grünwald-Giemsa-stained smears, most of the cells had reddish cytoplasm. Red to purple, myxoid matrix was present as a scanty fibrillar substance and as globules surrounded by tumor cells vaguely reminiscent of adenoid cystic carcinoma. A cytodiagnosis of myoepithelioma was given and corroborated by immunocytochemical staining, which revealed a positive reaction for vimentin, smooth muscle actin and S-100 protein. Epithelial membrane antigen yielded a negative reaction except for a few plasmacytoid cells with weakly positive staining. Histopathology of the resected tumor and immunohistochemical staining confirmed the cytodiagnosis of myoepithelioma. CONCLUSION: FNA cytologic features together with immunocytochemical studies on smears can offer a preoperative diagnosis of myoepithelioma.     

A Condensation-Ordering Mechanism in Nanoparticle-Catalyzed Peptide Aggregation

Nanoparticles introduced in living cells are capable of strongly promoting the aggregation of peptides and proteins. We use here molecular dynamics simulations to characterise in detail the process by which nanoparticle surfaces catalyse the self-assembly of peptides into fibrillar structures. The simulation of a system of hundreds of peptides over the millisecond timescale enables us to show that the mechanism of aggregation involves a first phase in which small structurally disordered oligomers assemble onto the nanoparticle and a second phase in which they evolve into highly ordered as their size increases.     

The relationships between ribosomal genes and fibrillar centers in thyroid cells cultivated in vitro

Despite the fact that the fibrillar centers of the nucleolus and the chromosomal nucleolar organisers (NORs) are similarly stained with the NOR-silver technique, there remain some questions about the identification of fibrillar centers as NORs. The distinct delineation of the fibrillar centers in porcine thyroid cells allowed us to determine whether there was a numerical equivalence or correlation between fibrillar centers and NORs. Hybridization in situ and silver staining performed on pig chromosomes showed that pairs 8 and 10 contained rDNA sites. Silver staining of thyroid cells in electron microscopy showed that the fibrillar centers and their surrounding layer of dense fibrils were the sites of silver deposit. Chromatin fibers were demonstrated within the fibrillar centers through the aid of the osmiumammine reaction and with the oxidized diaminobenzidine technique. It was observed that in cultured thyroid cells the fibrillar centers could be identified in the light microscope as argyrophilic spherules, and easily counted. The number of fibrillar centers was variable according to culture conditions. In cells cultured for 5 hr, the mean number of fibrillar centers was 1.7. After 5 days of culture, the number of fibrillar centers increased, reaching a mean value of 5.93. When thyroid cells were stimulated with thyrotropin, the number of fibrillar centers again increased to a mean value of 7.54. These results demonstrate that the relationship between fibrillar centers and NORs is not a simple proportionality: the number of fibrillar centers increases with increased cellular activity. These data imply that in active cells each NOR may pass through several fibrillar centers.     

Agrobacterium tumehciens After in vivo Inoculation of Potato (Sohnum tuberosum L.) (Scanning Electron Microscopy)

Five weeks after the in vivo inoculation of potatoes ( Solanum tuberosum L.) with Agrobacterium. tumefaciens strain B6S3, bacteria were found in the non-differentiated cells of tumors (formed from xylem parenchyma or other living cells), in xylem cells at the site of inoculation, as well as in xylem cells of the adjacent stem.
Bacteria were attached by fibrillar aggregates to the tumor cell walls. They were also attached to a fibrillar mass which arose from agrobacteria connected to this mass in the tumor. Agrobacteria, singly or in pairs, were attached to an electron dense formation (possibly bacterial extracellular polysaccharides) found both inside the xylem cells of the stem adjacent to the tumor and at the site of inoculation. Some A. tumefaciens cells were attached by means of a pedestal-like structure at the inoculation site.
A possible function of the different means of attachment of A. tumefaciens in both nontransformed plant cells and tumors is discussed.     

Dynamics and three-dimensional localization of ribosomal RNA within the nucleolus

Although rRNA synthesis, maturation, and assembly into preribosomal particles occur within the nucleolus, the route taken by pre-rRNAs from their synthetic sites toward the cytoplasm remains largely unexplored. Here, we employed a nondestructive method for the incorporation of BrUTP into the RNA of living cells. By using pulse-chase experiments, three-dimensional image reconstructions of confocal optical sections, and electron microscopy analysis of ultrathin sections, we were able to describe topological and spatial dynamics of rRNAs within the nucleolus. We identified the precise location and the volumic organization of four typical subdomains, in which rRNAs are successively moving towards the nucleolar periphery during their synthesis and processing steps. The incorporation of BrUTP takes place simultaneously within several tiny spheres, centered on the fibrillar centers. Then, the structures containing the newly synthesized RNAs enlarge and appear as compact ringlets disposed around the fibrillar centers. Later, they form hollow spheres surrounding the latter components and begin to fuse together. Finally, these structures widen and form large rings reaching the limits of the nucleoli. These results clearly show that the transport of pre-rRNAs within the nucleolus does not occur randomly, but appears as a radial flow starting from the fibrillar centers that form concentric rings, which finally fuse together as they progress toward the nucleolar periphery.     

Does the synthesis of ribosomal RNA take place-within nucleolar fibrillar centers or dense fibrillar components?

By means of immunocytochemistry performed on cryosections of cultured cells, RNA polymerase I was localized mainly to nucleolar fibrillar centers. The labelling of nucleolar dense fibrillar components was low and depended on the cell type. In contrast, DNA topoisomerase I and RNP complexes containing U3 snRNA were enriched in dense fibrillar components, their occurrence in fibrillar centers being usually much less.     

Fibrillar α-synuclein and huntingtin exon 1 assemblies are toxic to the cells

The aggregation of alpha-synuclein (α-syn) and huntingtin (htt) into fibrillar assemblies in nerve and glial cells is a molecular hallmark of Parkinson's and Huntington's diseases. Within the aggregation process, prefibrillar and fibrillar oligomeric species form. Prefibrillar assemblies rather than fibrils are nowadays considered cytotoxic. However, recent reports describing spreading of fibrillar assemblies from one cell to another, in cell cultures, animal models, and brains of grafted patients suggest a critical role for fibrillar assemblies in pathogenesis. Here we compare the cytotoxic effect of defined and comparable particle concentrations of on-assembly pathway oligomeric and fibrillar α-syn and Htt fragment corresponding to the first exon of the protein (HttEx1). We show that homogeneous populations of α-syn and HttEx1 fibrils, rather than their precursor on-assembly pathway oligomers, are highly toxic to cultured cells and induce apoptotic cell death. We document the reasons that make fibrils toxic. We show that α-syn and HttEx1 fibrils bind and permeabilize lipid vesicles. We also show that fibrils binding to the plasma membrane in cultured cells alter Ca(2+) homeostasis. Overall, our data indicate that fibrillar α-syn and HttEx1, rather than their precursor oligomers, are highly cytotoxic, the toxicity being associated to their ability to bind and permeabilize the cell membranes.     

Degradation of fodrin by m-calpain in fibroblasts adhering to fibrillar collagen I gel

When skin fibroblasts were cultured on fibrillar collagen I gel, we observed rapid degradation of talin, fodrin and ezrin, which are well-known calpain substrates. The protease m-calpain was activated only in cells adhering to fibrillar collagen, whereas micro-calpain was activated in cells adhering to monomeric or fibrillar collagen at the same level. The calpain inhibitor Z-Leu-Leu-aldehyde inhibited degradation of fodrin, but not talin. Degradation of fodrin, alpha-actinin and ezrin was prevented by over-expression of dominant negative m-calpain. However, over-expression of calpastatin, an endogenous calpain inhibitor, had no effect the degradation of these three proteins. These results suggest that m-calpain is responsible for degradation of their membrane proteins via adhesion to fibrillar collagen I gel.     

Morphology of healthy human parathyroid glands in cytologic smears

OBJECTIVE: To compare the morphology of different parenchymal cell types in healthy human parathyroid glands (HPGs) in cytologic smears with their structure on histologic sections and to establish criteria for their recognition in smears. STUDY DESIGN: The HPGs of 47 subjects (27 females and 20 males) were incidentally removed during surgery on the thyroid gland. The tissue of glands with a normal macroscopic and microscopic appearance was analyzed in cytologic smears and on histologic sections. RESULTS: In cytologic smears (as well on histologic sections), dark and light chief cells predominated. Dark chief cells, which were more numerous than light ones, had a smaller nucleus without a visible nucleolus. The nucleus of light chief cells was larger with 1 or 2 nucleoli visible. The cytoplasm of both types of chief cells was poorly defined, grey-blue and often vacuolated. In the smears, the cytoplasm of oxyphilic cells was dense, gray-rose and well defined. In dark oxyphilic cells, the nuclei were smaller and without a visible nucleolus. Light oxyphilic cells had a larger nucleus and visible nucleolus. CONCLUSION: The identification and distribution of 4 parenchymal cell subtypes in the smears of pathologically altered HPGs may yield insights into the possible role of these cells in a specific disease.     

3-dimensional organization of the nucleolus and the nucleolus-organizing regions in differentiated cells. I. Ring-shaped nucleoli in the endotheliocytes, smooth-muscle cells and fibroblasts in the mouse

The method of ultra-thin serial sections was used to study the three-dimensional structure and to perform the quantitative analysis of ring-shaped nucleoli of kidney and liver endotheliocytes, smooth muscle cells of kidney arterioles and fibroblasts of mice. Spatial models of ring-shaped nucleoli with one fibrillar centre are given. For the quantitative analysis the following parameters were measured: the number and volumes of nucleoli, fibrillar centers, RNP-containing structures, the vacuolar system and the RNP-index (the latter is a ratio of RNP-part and fibrillar center volumes). Nucleoli of the same type of cells, occasionally in the same nucleus, were found to differ sharply in their fibrillar center shape. Differences in the mean volume values of nucleoli, fibrillar centers and the RNP-part between some cell populations are sufficiently well pronounced. Within the same population ring-shaped nucleoli have, as a rule, specific volume values of nucleoli, RNP and fibrillar centers. The comparison of quantitative data obtained on different cell types showed that the mean RNP-index values were the most stable parameter. The structural relation between fibrillar centers, intra- and perinucleolar chromatin and lacunar region is shown. The structural organization of intranucleolar chromatin and rRNA in the nucleolar body and in fibrillar centers is discussed.     

Fine needle aspiration cytology of metastatic malignant schwannoma. A case report

A metastatic malignant schwannoma diagnosed by fine needle aspiration (FNA) biopsy in a 56-year-old man is reported. Cytologic examination of smears and cell blocks prepared from aspirates of a vertebral mass suggested the presence of metastases from a previously excised malignant schwannoma on the right leg. Electron microscopic and immunocytochemical studies on the aspirate supported the diagnosis, as did the patient's clinical history and previous pathology and the radiographic demonstration of metastatic lesions in the lung. The cytologic findings (cells with oval-to-spindled nuclei and ill-defined cellular borders suspended within a delicately fibrillar eosinophilic matrix) are discussed in light of the histologic diversity of this lesion and the problems of distinguishing it from other sarcomas. The ability to diagnose metastatic malignant schwannoma by FNA emphasizes the value of this technique.     

The erythroblastic island: past and future

The erythroblastic island consists of a macrophage surrounded by erythroblasts of different maturation stages. The cohesion of cells is lost during spreading, so that the islands can no longer be seen in smears. However, in undisturbed preparations, either living or fixed, the islands can be visualized, isolated, cultured, and their function studied. Many questions remain, such as the production of growth factors by the macrophage, the causation of the adhesion of cells forming the islands, the role of calcium in the adhesion, and others. New technical advances, including flow cytometry, immunolabeling, and in situ hybridization will assist in answering these questions.     

Human papillomavirus infection of the cervix: the atypical condyloma

We report on 162 cases of human papillomavirus (HPV) infection of the cervix seen in a two-year period in which the cell sample showed such marked atypia that errors of interpretation could easily have been made. These atypical condylomata are difficult to diagnose cytologically as well as histologically because they mimic dysplasia or carcinoma in situ and, on smears, even invasive squamous carcinoma. HPV particles associated with fibrillar material were found within nuclei of these lesions; their nature was further proved by the immunoperoxidase test. This new form of HPV infection of the cervix showed a 9.1% rate of progression to more advanced cervical lesions. The cytologic finding of atypical condylomata is an indication for colposcopy, confirmative biopsy and appropriate treatment.     

Nodular fasciitis: diagnosis by fine needle aspiration biopsy

OBJECTIVE: To describe the cytomorphologic features of nodular fasciitis that differentiate it from schwannoma. STUDY DESIGN: The cytomorphologic features of 10 cases of nodular fasciitis were compared to those of 4 cases of biopsy-proven schwannoma. Aspirate smears were evaluated for cellular cohesion, cell type and stroma. Immunoperoxidase stains were utilized in select cases. RESULTS: The cases of nodular fasciitis exhibited cohesive clusters of epithelioid to spindle-shaped cells in a background of single, intact mesenchymal cells; inflammatory cells; and myxoid stroma. In contrast, schwannomas lacked single, intact cells and inflammation. Schwannoma stroma was also myxoid but appeared more finely fibrillar, and cell clusters were notable for alternating areas of hypercellularity and hypocellularity. Immunoperoxidase stains demonstrated smooth muscle actin reactivity in 5 cases of nodular fasciitis and S-100 in 2 cases of schwannoma. CONCLUSION: Nodular fasciitis can be distinguished from schwannomas on the basis of cytomorphologic features and immunocytochemical profile. Cytologic diagnosis of nodular fasciitis is important since it obviates the need for surgical excision.     

Mesonephric hyperplasia can cause abnormal cervical smears: report of three cases with review of literature

BACKGROUND: Hyperplastic mesonephric remnants are an incidental finding in occasional uterine or cervical surgical specimens. We describe three cases in which such remnants were postulated to be the source of abnormal glandular cells in cervical smears. CASES: In all three cases abnormal glandular cells were seen in cervical smears. Subsequent histology showed the presence of hyperplastic mesonephric remnants that communicated with the endocervical canal and were likely to be the source of the abnormal glandular cells. We believe that the key features of these cells, which may aid their distinction from other causes of glandular abnormalities, are their loose clustering, lack of significant anisocytosis and cuboidal outlines. CONCLUSION: We aim to document mesonephric hyperplasia as a possible source for abnormal glandular cells in cervical smears.     

An immunogold-silver staining method for detection of cell surface antigens in cell smears

We developed an indirect immunogold-silver staining method for detection of leukocyte cell surface antigens in cell smears. Air-dried and fixed cytocentrifuge preparations or smears of peripheral blood leukocytes were incubated with monoclonal antibodies (MAb) and colloidal gold-labeled secondary antibodies. The preparations were post-fixed and silver enhancement was performed. The smears were counterstained with May-Grunwald-Giemsa and examined in brightfield light microscopy. The morphology of the cells was well preserved. Leukocytes reacting with the MAb showed black granules on their surface membranes. The intense immunostaining and the low background allowed a rapid enumeration of the positive cells. The labeling could be detected with high sensitivity by epipolarization microscopy. This immunogold-silver staining method was used to quantify T- and B-lymphocytes and natural killer cells in buffy coat smears of normal adult blood. These lymphocyte subsets correlated well with those obtained in smears with the alkaline phosphatase-anti-alkaline phosphatase (APAAP) method and with those found by labeling of mononuclear cells in suspension with immunogold-silver staining. This immunogold-silver staining method forms a good alternative to immunoenzyme methods for study of hematologic cells. In addition, it could be a general procedure for detection of cell surface antigens in all kinds of cell smears.     

Laser microdissection of actinomycin D segregated nucleoli

Nucleoli of tissue culture cells were segregated into their fibrillar (light) and granular (dark) components by treatment with actinomycin D. Following this segregation, the cells were treated with quinacrine hydrochloride, an agent which selectively sensitizes the nucleoli to argon laser light. The actinomycin D-segregated, quinacrine-sensitized nucleolar components (dark and light) were selectively irradiated with the laser microbeam and subsequent uridine uptake assayed. The data indicate that selective damage to the light (fibrillar) area is generally more damaging than damage to the dark (granular) area. These results support the idea that DNA is closely associated with the nucleolar fibrillar component.