Establishment of a human hepatocellular carcinoma cell line releasing hepatitis B virus surface antigen

A new hepatocellular carcinoma cell line, DELSH-5, derived from operative wedge biospy from a HBsAg sero- and tissue-positive patient, has been continuously propagated in vitro for nearly 22 months. The cells not only resemble hepatocytes on light and electron microscopic examination but also possess biosynthetic markers of the latter such as albumin and alpha-foetoprotein which were demonstrated in the supernatant medium as well as in the tumour cell cytoplasm. Karyology of cloned cells shows moderate aneuploidy, the major model chromosome number being 61. Though in the initial few passages HBsAg could not be detected, from the 13th passage onwards this viral component could be consistently demonstrated in small amounts in the concentrated supernatant medium by the macro- and micro-ELISA techniques. The immunohistochemical techniques as well as electron microscopy have failed to demonstrate any virus component inside the cell. The cell line reported here is the third of its kind which will act as a useful laboratory model to obtain pure HBsAg and to study the hepatitis-B-virus--liver-cell interaction with particular reference to the oncogenic potential of the virus.     

Cytopathology of uncommon malignant renal neoplasms in the pediatric age group

Malignant renal neoplasms are common solid tumors in pediatric oncology practice. These include the common Wilms' tumor/nephroblastoma and the uncommon neoplasms such as clear-cell sarcoma of the kidney (CCSK), rhabdoid tumor, renal-cell carcinoma, and others. The aim of this study was to describe in detail the cytopathological features of the histopathologically proven uncommon pediatric renal tumors. Aspirates from Wilms' tumor, which are mesenchyme predominant, show clusters of spindle cells associated with the matrix material. Evidence of rhabdomyoblastic differentiation may be present. CCSK, classic subtype, is characterized by round to oval cells arranged perivascularly and also in sheets and clusters intimately associated with a metachromatic matrix mucopolysaccharide material better appreciated in May-Grunwald-Giemsa (MGG)-stained smears. The cells also have more abundant cytoplasm and may show nuclear grooves. Spindle-cell pattern of CCSK is difficult to diagnose on aspiration cytology. Renal-cell carcinoma of childhood shows similar cytological features as its adult counterpart. Rhabdoid tumor of the kidney is characterized by a monomorphic population of cells with abundant cytoplasm, eccentric nuclei with prominent nucleoli. Intrarenal yolk sac tumor is a rare neoplasm and shows severely pleomorphic cells on aspiration.Awareness of these entities is important for the practicing cytopathologist. Further, non-Wilms' renal malignant neoplasms must be distinguished from the common Wilms' tumor so that appropriate chemotherapy protocols may be instituted in cases where the tumor is in an advanced stage of malignancy.     

Oxidant-mediated activation of cytosolic phospholipase a(2) in pulmonary endothelium: role of protein kinase C alpha and a pertussis toxin-sensitive protein.

The authors have previously demonstrated that the oxidant t-buOOH stimulates phospholipase A(2) (PLA(2)) activity in bovine pulmonary artery endothelial cells (S. Chakraborti et al. American Journal of Physiology, 257, L430-L437, 1989). Herein, the authors sought to investigate the mechanism by which t-buOOH stimulates PLA(2) activity and the role of protein kinase C (PKC) in this scenario. Treatment of bovine pulmonary artery endothelial cells with t-buOOH stimulated an aprotinin-sensitive protease activity, PKC activity, and PLA(2) activity in the cell membrane. Pretreatment with intracellular Ca(2+) chelator (BAPTA-AM), PKCalpha inhibitor (Go6976), cPLA(2) inhibitor (AACOCF(3)), and pertussis toxin prevented t-buOOH-stimulated PLA(2) activity. Immunoblot studies with aprotinin, cPLA(2), PKCalpha, and Gialpha antibodies revealed their presence in the endothelial membrane. Immunoblot studies of the cell membrane isolated from t-buOOH-stimulated cells with cPLA(2) and PKCalpha antibodies elicited an apparent increase in their immunoreactive protein profiles along with an additional 47-kDa immunoreactive fragment in the membrane. t-buOOH caused Gialpha phosphorylation in the membrane and pretreatment with Go6976 prevented the phosphorylation. Overall, these results suggest that t-buOOH stimulates an aprotinin-sensitive protease activity that proteolytically activates PKCalpha and that subsequently phosphorylates a pertussis toxin-sensitive protein, resulting in the stimulation of cPLA(2) activity in the cell membrane.     

Fine-needle aspiration biopsy in diagnosis of follicular lymphoma: cytomorphologic and immunohistochemical analysis

Follicular lymphoma is a relatively uncommon type of non-Hodgkin's lymphoma (NHL) thought to derive from follicular center cells. There are limitations to fine-needle aspiration in the diagnosis of follicular NHL, and very few studies are available on it. We describe here the cytomorphologic features of eight biopsy-proven cases of follicular NHLs and five biopsy-proven cases of intermediate-grade NHLs. Five cases of reactive lymphoid hyperplasia (RLH) diagnosed on cytology were also studied cytomorphologically. Image morphometry was done in follicular and intermediate-grade NHLs, immunocytochemistry was done in follicular and intermediate-grade NHLs, and immunocytochemistry for bcl-2 expression was studied in five cases of follicular NHLs. The aspirate smears of follicular NHL showed the presence of higher grades, many to abundant (++ to +++) monomorphic lymphoid aggregates, with an increased number of mast cells. Lymphohistiocytic tangles and tingeable body macrophages were not observed in any of the cases. Cytomorphologically, all the cases of intermediate-grade NHL showed a predominantly monomorphic population of lymphoid cells with absent or few grades (+) of lymphoid aggregates. Image morphometric data analysis of each cell parameter for follicular and intermediate-grade NHLs were found to be statistically insignificant (P > 0.05) and hence of little help in diagnosis of follicular vs. low-grade NHLs. Immunocytochemistry for bcl-2 expression in follicular NHL showed variable results, ranging from 0-90% (expressed as bcl-2 index). Certain characteristic cytomorphological features such as the presence of monomorphic lymphoid cell aggregates and an increased number of mast cells are soft indicators for the diagnosis of follicular NHL on aspiration smears. Immunocytochemistry for bcl-2 expression may be of help in cases with clinical and cytological suspicion of lymphoma. Image morphometry has a small role in this regard.     

HLA transgenic mice as models of human autoimmune diseases

MHC class II alleles have been linked to several human autoimmune diseases such as rheumatoid arthritis (RA), Type I diabetes, and multiple sclerosis (MS). Although the mechanisms by which expression of certain MHC class II molecules predispose an individual to a particular autoimmune disease are not known, it is clear that increased susceptibility is associated with the polymorphic regions unique to these predisposing HLA alleles. These polymorphic differences may influence susceptibility by selecting potential autoreactive T cells during thymic education. Alternatively, nonsusceptible alleles may either delete or fail to select these potential autoimmune T cells, thus reducing the possibility of developing disease. In the periphery, the unique specificity of the HLA molecule derived from a susceptible allele may then recognize and present an autoantigenic peptide or foreign peptide that may cross-react with an autoantigen, activating these autoreactive T cells and leading to disease. To dissect these possibilities and to determine the exact role of particular HLA-DR or DQ molecules in disease susceptibility, we have generated several lines of HLA-DR and DQ transgenic mice. In this review, we present data summarizing the functions of these HLA class II molecules using well-established mouse models for autoimmune diseases.     

Modulation transfer functions of tomographic systems

The modulation transfer function for an elliptical tomographic scan is derived. Its imaging characteristics are compared with those of linear and circular scans

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Zusammenfassung Die Modulationsaustauschfunktion einer elliptischen tomographischen Abastung wird abgeleitet. Ihre Abbildungsmerkmale werden mit denen von linearen und runden Abtastungen verglichen.

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Sommaire La fonction de transfert de modulation d'un balayage tomographique elliptique est dérivée dans cet article. Ses caratéristiques d'image sont comparées avec celles des balayages circulaires et linéaires.

     

Microstructure and deformation behavior of biocompatible TiO2 nanotubes on titanium substrate

Titanium oxide coatings have been shown to exhibit desirable properties as biocompatible coatings. We report on the quantitative microstructure characterization and deformation behavior of TiO₂ nanotubes on Ti substrate. Nanotubes were processed using anodic oxidation of Ti in a NaF electrolyte solution. Characterization of the as-processed coatings was conducted using scanning electron microscopy and focused ion beam milling. Increases in anodization time had no significant effect on tube diameter or tube wall thickness. Coating thickness, however, increased with time up to 2 h of anodization, at which point an equilibrium thickness was established. Nanoindentation was used to probe the mechanical response in terms of Young’s modulus and hardness. Progressively higher values of elastic modulus were obtained for thinner films consistent with increasing effects of the Ti substrate. A possible deformation mechanism of densification of the porous oxide and wear of the dense surface is suggested and discussed.     

Diagnosis of retinoblastoma by fine-needle aspiration and aqueous cytology

This article reports the cytodiagnosis of three cases of retinoblastoma in children aged 1.5, 2.5, and 5 yr. Two of them were diagnosed by fine-needle aspiration cytology of the primary tumor and one by aqueous cytology. The tumor cells were usually round to oval, small and uniform, with scanty cytoplasm; they generally occurred in closely packed clusters of variable sizes. We discuss the differential diagnosis of retinoblastoma with other round-cell tumors of childhood involving the orbit.     

Isolation and preliminary characterization of Escherichia coli mutants resistant to lethal action of the bacteriophage lambda P gene.

Both spontaneous and NTG-induced mutants of Escherichia coli 594 insensitive to the lethal action of lambda P gene were isolated and called rpl (resistant to P lethality). These mutants were of two types, showing different phenotypes. On type I rpl mutants, lambda cl- and lambda v1v3 did not plate, while lambda vir, lambda cl- c17, lambda imm434, and lambda imm21 did; plasmid pMR45 carrying the lambda P gene could not complement lambda imm21P- phage in type I mutants. On the other hand, the type II rpl mutants support the growth of all the above phages including lambda cl-. Neither type of rpl mutation affects growth of the bacteria.