Nelly Livni

Proliferative activity and p53 expression in primary and recurrent pterygia

PURPOSE To assess p53 expression and proliferative activity in primary and recurrent pterygia from the same eyes. DESIGN Retrospective comparative human tissue study. PARTICIPANTS Tissue from excised primary pterygia that did not recur (group A, n = 10) was compared with tissue from primary pterygia that recurred (group B, n = 10) and to the recurrent pterygia tissue that was excised from subjects in group B (group C, n = 10). Ten normal conjunctivas served as controls (group D). METHODS Sections from each pterygium were immunostained with the MIB-1 and bp53. 12 monoclonal antibodies that react with Ki-67 and p53 antigens, respectively. MAIN OUTCOME MEASURES Proliferative activity was calculated as the mean of the MIB-1 positive cell count per eyepiece grid in high magnification (x40) (positive cell count/grid). Percentage of positive cells of all cells in the grid area was evaluated in the p53-stained sections. RESULTS Proliferative activity was found in the epithelium overlying the pterygia and normal conjunctiva. The mean MIB-1 positive cell count/grid +/- standard error was 2.84 +/- 1.07, 1.74 +/- 0.82, 3.83 +/- 1.35, and 0.86 +/- 0.33 in groups A, B, C, and D, respectively (P = 0.17, Kruskal-Wallis). P53 staining was found in 50% of pterygia in groups A, B, and C; none of the normal conjunctival tissues showed p53 immunoreactivity. Four of five p53-positive tissues in group B were p53-negative in group C. In the p53-positive pterygia, less than 10% of cells were p53 positive. However, p53-positive pterygia had higher mean MIB-1 positive cell count/grid +/- standard error as compared with the p53-negative lesions, 4.56 +/- 0.94 vs 1.39 +/- 0.59 (P = 0.021, Mann-Whitney). CONCLUSIONS p53 immunoreactivity and high proliferative activity in the epithelium overlying the pterygium are not associated with recurrence of pterygium.

Systemic lupus erythematosus associated with acute Epstein-Barr virus infection

Systemic lupus erythematosus (SLE) is a multisystem disease of unknown origin, characterized by a variety of autoimmune phenomena. Viruses have long been postulated to play a role in its pathogenesis. Several observations suggested a link between Epstein-Barr virus (EBV) and SLE. We describe a 14-year-old girl who presented with acute onset of SLE concurrently with clinical and laboratory findings consistent with EBV-induced infectious mononucleosis (IM). Evidence for acute EBV infection was confirmed by serological studies and detection of specific EBV antigens on kidney biopsy. This close association between EBV and SLE suggests a possible role of the virus in the pathogenesis of SLE in this patient.

P53 expression in patients with cirrhosis with and without hepatocellular carcinoma

Background. Mutated p53 acts as a dominant oncogene, whereas the wild type (wt) p53 gene product suppresses cell growth. Abnormalities in the p53 gene are reported in more than 50% of malignant tumors. Recently, an allelic loss of chromosome 17p, where the p53 gene is located, was found to be more frequent in hepatocellular carcinoma (HCC) cell lines and human tumors. In addition, in half of the cases of HCC from endemic areas for hepatitis B virus and aflatoxin, a hot spot point mutation at codon 249 was detected, as previously reported. Missense mutations in p53, mdm‐2 complex formation, and other unknown mechanisms may lead to stabilization of the gene product, thus rendering it detectable by immunohistochemistry.

PC-10 Immunostaining of Proliferating Cell Nuclear Antigen in Posterior Uveal Melanoma: Enucleadon versus Enucleation Posdrradiadon Groups

PURPOSE It is difficult to assess the viability of uveal melanoma after radiotherapy treatment. The purpose of the current study is to investigate PC-10 monoclonal antibody of proliferating cell nuclear antigen as a possible marker for cell proliferation and tumor viability in conventionally processed histologic preparations of uveal melanoma irradiated by brachytherapy as well as in nonirradiated melanomas. METHODS Thirteen enucleated eyes with posterior uveal melanoma that were treated by brachytherapy (cobalt 60 or ruthenium 106 radioactive plaques) were included in this study. Thirteen enucleated eyes of the same size with nonirradiated posterior uveal melanoma served as controls. The tumors were stained with PC-10 monoclonal antibody to proliferating cell nuclear antigen. All clinical and histologic data of the tumors were recorded and analyzed. RESULTS Five of the irradiated tumors showed positive staining with PC-10, although with a low score. Four of these tumors showed regrowth, and the fifth tumor was treated with a low-irradiation dose (5500 rad). In the nonirradiated tumor group, nine were positive for PC-10 staining, with a higher score. Significant correlation was found in this group between the PC-10 score and the mitotic figure count, but not with other prognostic factors. In three of four tumors that caused metastatic death, the PC-10 staining was positive and had a high score. CONCLUSIONS PC-10 immunostaining is a simple, reproducible method that can be applicable to conventionally processed histologic preparations. It clearly shows that cellular proliferation activity in nonirradiated and irradiated uveal melanomas. Based on the small number of cases reported herein, it seems that the PC-10 score can correlate with prognosis, but further studies should be performed.

p53 Immunoreactivity, Ki-67 expression, and microcirculation patterns in melanoma of the iris, ciliary body, and choroid

Purpose. The major role of p53 is to modulate cell proliferation, but recently, it has been found that p53 also modulates angiogenesis through several pathways. Because both cellular proliferation and microcirculation patterns are important prognostic markers in uveal melanoma, we tested the relationships between p53 expression, the expression of the cell proliferation marker Ki-67, and the presence of various microcirculation patterns in uveal melanoma. Methods. Immunostaining of p53 and Ki-67 using the bp53.12 and the MIB-1 antibodies, respectively, were preformed in 98 uveal melanomas (18 melanomas confined to the iris, 30 ciliary body melanomas, and 50 choroidal melanomas). Percent of p53 positive cells, and the mean MIB-1 positive cell count per high power field were calculated in each section by two observers. Microcirculation patterns were assessed in adjacent PAS stained sections. Results. p53 immunoreactivity was found in 14 of the 98 melanomas. High proliferative activity and epithelioid cell type were associated with p53 immunoreactivity. However, p53 immunoreactivity was not associated with any of the microcirculation patterns or with tumor location. Conclusions. The findings suggest that alterations in p53 expression are associated with the expression of the cellular proliferation marker, Ki-67, but are not associated with the presence of microcirculation patterns.

Induction of lipid storage in cultured leukemic myeloid cells by pyrene-dodecanoic acid

When incubated for 1-3 days in the presence of the fatty acid analog, 12-(1-pyrene)dodecanoic acid, the neutral lipid content of cultured human leukemic myeloid cells increased considerably, while that of the phospholipids increased to a much lesser extent. Among the neutral lipids, di- and monoacylglycerols predominated and a considerable portion of the fatty acyl residues of these newly synthesized neutral lipids consisted of pyrene-dodecanoic acid. Light microscopy showed evidence for the presence of highly fluorescent lipid droplets within the cells. Electron microscopy showed lipid globules, mostly devoid of a unit membrane, multivesicular inclusion bodies and some multilamellar membranous structures. In comparison, cells incubated with palmitic acid show neither these cellular structures, nor the increase of the neutral lipid content. The lipid storage, induced by pyrene-dodecanoic acid, is probably related to ineffective degradation of this fatty acid analog and might serve as an experimental model of cellular lipidosis.

Immunoperoxidase method of identification ofLeishmania in routinely prepared histological sections

An indirect immunoperoxidase technique was applied for identification ofleishmania in routinely prepared histological sections. Paraffin embedded, formalin-fixed slide preparations of skin (1 case), bone marrow (2 cases) and lymph node (1 case) were examined. The tissues were obtained from one patient with cutaneous leishmaniasis and from three patients with visceral leishmaniasis. Specific rabbit anti-sera againstL. donovani, L. tropica andL. mexicana were used as primary reagents. Positive controls were performed simultaneously and includedL. tropica cultures in blood-agar (NNN media) - free promastigotes and amastigotes within macrophages. Strongly positive brown staining was localized specifically in Leishman-Donovan (LD) bodies only. This method increases the probability of microscopic diagnosis of leishmaniasis and helps to prevent confusion ofLeishmania with other infective agents in histological sections.

Multiple hornet stings with features of Reye's syndrome.

Hornets venom is known to possess a variety of toxic effects. A 19-mo-old girl who developed a Reye-like syndrome following multiple stings by the Oriental hornet (Vespa orientalis) is described. She presented with encephalopathy associated with hepatomegaly, elevated transaminase levels, low prothrombin time, and hyperammonemia. Liver biopsy demonstrated microvesicular fatty infiltration and diffuse mitochondrial changes. Additional features were acute renal tubular necrosis and massive hemolysis.

Comparison of microcirculation patterns and MIB-1 immunoreactivity in iris and posterior uveal melanoma

PURPOSE To compare melanomas confined to the iris and those involving either the ciliary body or choroid for the histologic features of microcirculation patterns and tumor cell proliferation indices. DESIGN Retrospective comparative human tissue study. PARTICIPANTS Ninety-eight uveal melanomas were studied, including 18 tumors confined to the iris, 30 tumors involving the ciliary body, and 50 tumors confined to the choroid. METHODS Formalin-fixed, paraffin-embedded sections from each tumor were stained with hematoxylin-eosin and with periodic acid-Schiff. Adjacent histologic sections were stained with the MIB-1 antibody that reacts with the Ki-67 antigen. MAIN OUTCOME MEASURES Microcirculation patterns were assessed in the periodic acid-Schiff-stained sections. Proliferative activity was assessed in the MIB-1-stained sections. The mean MIB-1 positive cell count per high-power field (HPF) was calculated in 10 HPF (x 40) in the area of maximal immunoreactivity. Two observers evaluated each MIB-1-stained section, and the interobserver reproducibility was assessed. RESULTS Histologic microcirculation patterns associated with death from metastatic disease in ciliary body and choroidal melanomas (parallel vessels with cross-linking and networks of back-to-back loops) were not found in any of the iris melanomas. By contrast, 34% and 63% of the choroidal and ciliary body melanomas, respectively, showed at least one of these patterns. The mean positive cell count per HPF +/- standard error was 19.9 +/- 3.5, 27 +/- 5.3, and 1.9 +/- 0.4 in choroidal, ciliary body, and iris melanoma, respectively (P: = 0.003, Kruskal-Wallis test). CONCLUSIONS Melanoma confined to the iris is characterized by a low rate of proliferation and the histologic absence of microcirculation patterns associated with metastatic posterior uveal melanoma. Both features are consistent with the relatively benign nature of iris lesions compared with melanomas involving the ciliary body or choroid.

Farage, a Novel Early B Cell Lymphoma Cell Line with Trisomy 11

Farage, a new cell line established from a lymph node biopsy of a patient with non-Hodgkins lymphoma (NHL), constitutes a clonal expansion of cells at a distinct stage of B-cell differentiation. The cells lack both T and myeloid surface markers, express B cell surface antigens including CD19, CD20, CD22, HLA-DR, were positive for C3 receptors and EBNA and expressed BCL-2. No immunoglobulin determinants could be demonstrated on the cell surface. Intracellular IgM and kappa chains were detected, in an unusual but distinct localization and appeared to be localized to the nucleus or to the perinuclear area without any spread to the cytoplasm, as seen in the early B cells. Southern blot DNA analysis showed rearrangement of one of the IgJH alleles. The Farage cells were negative for B cell activation antigens including CD25, CD11b, HC2 and Bly-7. The cells were negative for two anti CD-10 (CALLA) reagents but weakly positive with one. Interestingly they were strongly positive for both nuclear and cytoplasmic T...