Establishment and characterization of cell lines derived from nude mice transplanted squamous cell carcinoma of uterine cervix

Two human cell lines, KIMI-1 and -2 were established from nude mice transplanted tumor originated from a human squamous cell carcinoma of the uterine cervix. These two cell lines have different shapes, chromosome numbers and tumor markers, respectively.     

Simplified method for the chemical diagnosis of organic aciduria using GC/MS

GC/MS is widely used for the analysis of urinary organic acids for the chemical diagnosis of organic acidurias such as methylmalonic acidemia, propionic acidemia, isovaleric acidemia, glutaric aciduria type I, and multiple carboxylase deficiency. In this study, a rapid and simple preparation method for this analysis was developed in order to improve the laboratory productivity and the working environment. The solvent extraction and trimethylsilyl derivatization steps of the conventional method were improved by reducing the volume of urine sample and extraction solvent and by applying the flash-heater derivatization, respectively. The new method was successfully applied to the chemical diagnoses of five organic acidurias.     

The relation between recalcitrancy of a mangrove plant, Kandelia obovata, and high endogenous level of abscisic acid

Kandelia obovata Sheue, Liu & Yong sp. nov. is one of the cold tolerant mangrove plants. Some callus formation was obtained from the leaves of K. obovata in liquid medium containing 10 μM of 2,4-dichlorophenoxyacetic acid. However, recalcitrancy was found when subculturing them. Endogenous levels of gibberellins (GAs) and abscisic acid (ABA) in leaf protoplasts of K. obovata were determined using micro-scale extraction and purification steps, including thin layer chromatography and quantification by micro-bioassay or enzyme linked immunosorbent assays. Very high amounts of ABA and low activities of GAs were found in leaf protoplasts of K. obovata. Low concentrations of gibberellic acid and uniconazole-P were effective of enhancing cell enlargement in protoplast cultures. Exogenous application of ABA was inhibitory to protoplast culture. All cytokinins tested were inhibitory to both leaf and protoplast cultures. The high endogenous level of ABA is most likely the underlying cause of recalcitrancy of mangrove cultures.     

An Epidermal Growth Factor-like Repeat of Del1 Protein Increases the Efficiency of Gene Transfer In Vitro

In this paper, we found that Del1, an extracellular matrix protein secreted by embryonic endothelial cells, increases the efficiency of transfection in vitro. Conditioned medium containing Del1 increased transfection by the LacZ gene using several non-viral gene transfer systems, including lipoplex and polyplex methods. Experiments using deletion mutants and fragments of Del1 revealed that the third epidermal growth factor-like repeat (E3) of Del1 mediates the enhancement of gene transfer and, furthermore, that the motif CXDXXXFXCXC is essential. Incubation of Pro5 cells, a yolk sac-derived cell line, with as low as 16 pM recombinant E3 was sufficient to enhance transfection, and 1 nM recombinant E3 enhanced the transfection 12-fold. Inhibitors of endocytosis suppressed this activity of the recombinant E3. These results suggest that the E3 fragment of Del1 can be used as a general biological enhancer of non-viral gene transfer.     

Diagnostic usefulness of endometrial aspiration cytology for endometrial cancer cases with normal curettage findings

OBJECTIVE: To determine whether endometrial aspiration cytology is useful for endometrial cancer cases with normal endometrial curettage findings. STUDY DESIGN: Eleven cases in which endometrial cancer could be detected by endometrial aspiration cytology but not endometrial curettage were classified into 2 groups by cancer locus, on the endometrial surface (A) or in the myometrium (B). A clinicopathologic and cytologic analysis was performed to compare the 2 groups. RESULTS: Five cases had cancer lesions localized at the fundus and one at the isthmus (group A). The other 5 had lesions localized in the myometrium (group B). The myometrium invasion was beyond half the myometrium in group B and within half in group A. It required > 2 cytologic examinations for a definitive diagnosis in 33.3% of group A and 80.0% of group B. The endometrial cytology differed clearly between the groups: large clusters of malignant cells with a dirty background (group A) vs. small clusters with a clean background (group B). The log-rank test revealed that group B had significantly poorer prognoses than did group A despite nearly the same rate of stage I/II cases in the 2 groups (p = 0.004). CONCLUSION: Endometrial aspiration cytology was useful for endometrial cancer cases with normal curettage findings as part of early detection. However, the cytologic diagnosis did not indicate good prognoses in the cases of cancer localized in the myometrium.     

Selective activation of vitamin D receptor by lithocholic acid acetate, a bile acid derivative

The vitamin D receptor (VDR), a member of the nuclear receptor superfamily, mediates the biological actions of the active form of vitamin D, 1alpha,25-dihydroxyvitamin D(3). It regulates calcium homeostasis, immunity, cellular differentiation, and other physiological processes. Recently, VDR was found to respond to bile acids as well as other nuclear receptors, farnesoid X receptor (FXR) and pregnane X receptor (PXR). The toxic bile acid lithocholic acid (LCA) induces its metabolism through VDR interaction. To elucidate the structure-function relationship between VDR and bile acids, we examined the effect of several LCA derivatives on VDR activation and identified compounds with more potent activity than LCA. LCA acetate is the most potent of these VDR agonists. It binds directly to VDR and activates the receptor with 30 times the potency of LCA and has no or minimal activity on FXR and PXR. LCA acetate effectively induced the expression of VDR target genes in intestinal cells. Unlike LCA, LCA acetate inhibited the proliferation of human monoblastic leukemia cells and induced their monocytic differentiation. We propose a docking model for LCA acetate binding to VDR. The development of VDR agonists derived from bile acids should be useful to elucidate ligand-selective VDR functions.