Induction of a differentiated ciliated cell phenotype in primary cultures of Fallopian tube epithelium

Human Fallopian tubal epithelial cells in culture lose morphological features associated with the epithelium in situ and the extent to which they retain their in-vivo phenotype or function is unknown. In order to address this question, immunocytochemical markers were identified which distinguish secretory (HMFG2+, LhS28-) from ciliated (HMFG2-, LhS28+) epithelial cells in tissue sections of Fallopian tube. These markers were used to analyse the phenotype of tubal cells in vitro. Primary cultures of human tubal epithelial cells were seeded onto glass and grown to confluence before addition of oestradiol-17beta. In the absence of hormone, tubal epithelial cells expressed cytokeratins and nuclear receptors for oestrogen and progesterone and adopted a homogeneous (HMFG2+, LhS28-) secretory cell phenotype. Following the addition of oestradiol-17beta, a proportion of cells became positive for LhS28. The induction of a ciliated epithelial cell phenotype was confirmed by scanning electron microscopy, where on permeable collagen membranes, approximately one-third of tubal epithelial cells became ciliated in the presence of oestradiol-17beta. We suggest that in vitro, tubal epithelial cells adopt an immature secretory-like phenotype and that oestrogen can induce differentiation to a ciliated epithelial cell phenotype.      

Characterization of cell lines derived from hamster tumors induced with the BK virus

Summary Some of the properties of three continuous cell lines derived from BK virus-induced hamster tumors were examined. The cell lines hadin vitro growth characteristics of transformed cells. Morphologically most of the cells were fibroblastic, but multinucleated giant cells were also common. Ultrastructurally all three cell lines displayed the usual features of cells grownin vitro. Marked variation in the nuclear size and shape as well as prominent nucleoli were characteristic to these cells. No viruses or virus-like particles were found. Virus isolation attempts by fusing the cells with Vero cells were negative, and no virion antigen was detected in these cells by immunofluorescence. T antigen similar to that of other papovaviruses was found in the cells. This antigen stained with sera from a number of hamsters carrying transplanted BK virus-induced tumors, and also with SV 40 T antisera. The antigen disappeared after 30 minutes at 56° C. Cytogenetic analyses showed that the three cell lines were heteroploid with subtetraploid numbers of chromosomes. Chromosome abnormalities were also seen. All three cell lines induced sarcomatous tumors in adult hamsters after subcutaneous inoculation.With 4 Figures     

Absence of group II phospholipase A2, a Paneth cell marker, from the epididymis

Paneth cell-like metaplasia has been reported in the epithelium of the epididymis and prostatic adenocarcinomas. We studied the expression of group II phospholipase A2 (PLA2), a marker of Paneth cell differentiation, in six orchiectomy specimens with Paneth cell-like metaplasia. Both immunohistochemistry for group II PLA2 protein and in situ hybridization for the mRNA of group II PLA2 gave negative results in all six cases but positive reaction for lysozyme. The results show that the cells of the Paneth cell-like metaplasia are not true Paneth cells.     

Poor survival of cementless Biomet total hip: a report on 1,047 hips from the Finnish Arthroplasty Register

In Finland, almost 50% of all hip replacements done after 1989 have been inserted without cement. Biomet components have been the most commonly used implants in cementless arthroplasties. Between 1985 and 1997, 4,300 prostheses were implanted because of primary osteoarthrosis. 4 different acetabular component designs have been identified as Biomet implants (Mallory-Head, Romanus, T-TAP, Universal) and were used in 1,047 hips. The 9-year survival of all arthroplasties using Biomet cups was only 65 (95% CI 61-69)% while that of arthroplasties using T-TAP-cups was only 58 (52-65)%. In contrast, the 7.5-year survival of arthroplasties using Romanus cups was 85 (79-91)%. The 98 (96-99)% 5-year survival of arthroplasties with Mallory-Head cups should be interpreted cautiously, since similar results of arthroplasties using the Universal cup with the same type of liner decline sharply to 93 (88-98)% only 1 year later. The poor survival of Biomet cementless prostheses in our series seems to be related to the poor survival of the cup. This finding was common to all metal shell designs using Hexloc liners. We recommend that Biomet cups with Hexloc liners should not be used and patients who have had them inserted should undergo regular clinical and radiographic follow-ups.     

Anandamide prodrugs. 1. Water-soluble phosphate esters of arachidonylethanolamide and R-methanandamide.

Phosphate esters of arachidonylethanolamide (AEA) and R-methanandamide were synthesized and evaluated as water-soluble prodrugs. Various physicochemical properties (pK(a), partition coefficient, aqueous solubility) were determined for the synthesized phosphate esters. The chemical stability of phosphate esters was determined at pH 7.4. In vitro enzymatic hydrolysis rates were determined in 10% liver homogenate, and in a pure enzyme-containing (alkaline phosphatase) solution at pH 7.4. The intraocular pressure (IOP) lowering properties of R-methanandamide phosphate ester were tested on normotensive rabbits. The phosphate promoiety increased the aqueous solubility of the parent compounds by more than 16500-fold at pH 7.4. Phosphate esters were stable in buffer solutions, but rapidly hydrolyzed to their parent compounds in alkaline phosphatase solution (t(1/2)<15 s) and liver homogenate (t(1/2)=8-9 min). The phosphate ester of R-methanandamide reduced IOP in rabbits. These results indicate that the phosphate esters of AEA and R-methanandamide are useful water-soluble prodrugs.     

Hemoglobin and albumin adducts of benzene oxide among workers exposed to high levels of benzene

Benzene oxide (BO) reacts with cysteinyl residues in hemoglobin (Hb) and albumin (Alb) to form protein adducts (BO-Hb and BO-Alb), which are presumed to be specific biomarkers of exposure to benzene. We analyzed BO-Hb in 43 exposed workers and 42 unexposed controls, and BO-Alb in a subsample consisting of 19 workers and 19 controls from Shanghai, China, as part of a larger cross-sectional study of benzene biomarkers. The adducts were analyzed by gas chromatography-mass spectrometry following reaction of the protein with trifluoroacetic anhydride and methanesulfonic acid. When subjects were divided into controls (n = 42) and workers exposed to < or =31 (n = 21) and >31 p.p.m. (n = 22) benzene, median BO-Hb levels were 32.0, 46.7 and 129 pmol/g globin, respectively (correlation with exposure: Spearman r = 0.67, P < 0.0001). To our knowledge, these results represent the first observation in humans that BO-Hb levels are significantly correlated with benzene exposure. Median BO-Alb levels in these 3 groups were 103 (n = 19), 351 (n = 7) and 2010 (n = 12) pmol/g Alb, respectively, also reflecting a significant correlation with exposure (Spearman r = 0.90, P < 0.0001). The blood dose of BO predicted from both Hb and Alb adducts was very similar. These results clearly affirm the use of both Hb and Alb adducts of BO as biomarkers of exposure to high levels of benzene. As part of our investigation of the background levels of BO-Hb and BO-Alb found in unexposed persons, we analyzed recombinant human Hb and Alb for BO adducts. Significant levels of both BO-Hb (19.7 pmol/g) and BO-Alb (41.9 pmol/g) were detected, suggesting that portions of the observed background adducts reflect an artifact of the assay, while other portions are indicative of either unknown exposures or endogenous production of adducts.