Satoko Yamashita

Epidermal growth factor increased the expression of α2β1-integrin and modulated integrin-mediated signaling in human cervical adenocarcinoma cells

Epidermal growth factor (EGF) receptor (EGFR) is involved in various basic biochemical pathways and is thus thought to play an important role in cell migration. We examined the effect of EGF on motility, migration, and morphology of a human adenocarcinoma cell line CAC-1. EGF treatment increased the motility of cervical adenocarcinoma cells and promoted migration of the cells on fibronectin and type IV collagen. EGF induced morphological changes with lamellipodia during EGFR-mediated motility. The results of an immunoprecipitation study showed that EGF up-regulated the expression of alpha2beta1-integrin in a dose-dependent manner. EGF-induced cell migration was blocked by alpha2beta1-integrin antibody. Our results also showed that EGF treatment stimulated the level of tyrosine dephosphorylation of FAK, which is required for EGF-induced changes in motility, migration, and cell morphology. A tyrosine kinase inhibitor (ZD1839) blocked EGF-induced changes in cervical adenocarcinoma cells. The results suggest that EGF promotes cell motility and migration and increases the expression of alpha2beta1-integrin, possibly by decreasing FAK phosphorylation.

A 28 kDa protein of the Bacillus thuringiensis serovar shandongiensis isolate 89-T-34-22 induces a human leukemic cell-specific cytotoxicity

A 28 kDa protein that exhibits cytocidal activity specific for human leukemic T (MOLT-4) cells was purified from proteinase K-digested parasporal inclusion of a Bacillus thuringiensis serovar shandongiensis isolate. The N-terminal sequence of the protein was identical with that of the 32 kDa protein, regarded as a protoxin, of the inclusion proteins. The median effective concentration of this protein was 0.23 microg/ml against MOLT-4 cells and its specific activity was 7.9 times greater than that of the whole inclusion proteins. The 28 kDa protein induced necrosis-like cytotoxicity against MOLT-4 cells and the cytopathic effect with the passage of time was characterized by cell swelling, nuclear membrane isolation and chromatin condensation.

Isolation of Bacillus thuringiensis from intertidal brackish sediments in mangroves

Intertidal brackish sediments in mangroves were examined for isolation of Bacillus thuringiensis strains with novel toxicity spectra. A total of 18 B. thuringiensis isolates were recovered from eight sediment samples (36.4%) out of 22 samples tested. The frequency of B. thuringiensis was 1.3% among the colonies of Bacillus cereus/B. thuringiensis group. While five isolates were allocated to the four H serogroups, the majority of the isolates were serologically untypable or untestable. Two isolates belonging to the serovar israelensis/tochigiensis (H14/19) exhibited strong toxicities against larvae of the mosquito, Culex pipiens molestus, and mammalian cells (sheep erythrocyte and two human cancer cell lines) in vitro. The other 16 isolates showed no toxicity against the mosquito and mammalian cells. None of the isolates showed larvicidal activity against the diamondback moth, Plutella xylostella. Strong lectin activities against sheep erythrocytes were associated with two serologically untestable isolates and an H3 isolate.

Bacillus thuringiensis: a common member of microflora in activated sludges of a sewage treatment plant.

Bacillus thuringiensis was recovered at a high frequency from activated-sludge system environments in an urban sewage-digestive plant. All of the test materials, sampled at several digesting steps, contained the organism. Of 515 colonies belonging to the B. cereus/B. thuringiensis group, 45 (8.7%) were assigned to B. thuringiensis. The highest density of this bacterium was 1.6 × 103 cfu/ml in a scum sample of the first aeration basin. Among the 45 isolates, 7 were assigned to the known H serovars. Two isolates of the serovar kenyae isolates exhibited Lepidoptera-specific toxicity. Diptera-specific toxicity was shown by an isolate of serovar israelensis and a serologically undefined isolate. Lectin activity was associated with 12 isolates.

Cloning and characterization of two novel crystal protein genes from a Bacillus thuringiensis serovar dakota strain.

Two genes encoding the 32- and 40-kDa polypeptides of Bacillus thuringiensis strain 90-F-45-14 crystals were cloned, expressed in an acrystalliferous B. thuringiensis strain, and sequenced. The polypeptides had deduced molecular weights of 30,319 and 33,885, respectively. The amino acid sequence of the 32-kDa protein was 37.7% identical to the known sequence of a non-insecticidal parasporal protein in B. thuringiensis serovar thompsoni crystals. The sequence of the cloned 40-kDa protein was 37.0% and 22.3% identical to that of the existing Cry protein classes, Cry15Aa1 and Cry23Aa1, respectively. Thus, this protein constitutes a novel protein class, Cry33Aa1. The open reading frames of the two genes were located on the predominant plasmid of 17,629 bp (=11,752 MDa) in the same orientation, and they were separated by the sequence of 32 nucleotides. The two proteins are likely produced simultaneously from a single transcript to form spherical crystals.

HMB-45 staining for cytology of primary melanoma of the vagina. A case report.

BACKGROUND Malignant melanoma in the vagina is very rare, but its diagnosis is usually easy if a melanin pigment is present. With cytodiagnosis, however, it is difficult to differentiate amelanotic melanoma or scantily pigmented melanoma from other conditions. In the present case, monoclonal antibody HMB-45, the efficacy of which has been established in histologic studies, was used in the cytodiagnosis of amelanotic melanoma in the vagina. CASE A woman, aged 78 years, presented with a brownish, nodular tumor, diameter 3 cm, in the vagina. Scraping smears with Papanicolaou staining showed nonepithelial malignant cells without granules suggesting melanin. Smears stained with HMB-45 showed positive immunoreactivity. The diagnosis underwent histologic confirmation of amelanotic melanoma on the initial biopsy. CONCLUSION Cytodiagnosis was made with HMB-45, which proved very effective in the differential cytodiagnosis of amelanotic melanoma and scantily pigmented melanoma, particularly because it obviated the need for tissue invasion.

Surgical Management of Genital Prolapse: Is Chain Cystourethrography Useful for Evaluating Anatomical Corrections and Urinary Symptoms after Surgery?

Objective: To evaluate the relationship between chain cystourethrography and surgery for genital prolapse and urinary symptoms.

Characterization of Subclonal Cell Lines Showing High Motility Derived from an Undifferentiated Human Endometrial Cancer Cell Line

To examine the relationship between the morphology and the motility of cancer cells, we studied the difference between human endometrial undifferentiated cancer cell lines with different motilities, using light and electron microscopy, immunocytochemistry and RT-PCR. In a transplanted tumor of a cell line with high motility, many connective tissues and microvessels were observed and there were few intercellular spaces, which were ultrastructurally full of extracellular matrix. In the high-motility cell line only cytokeratin 19 was observed, and the level of cytokeratin 19 mRNA was elevated. Thus, motility is suggested to be related to the induction of connective tissues and cytoskeleton.