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The expression of the murine double minute-2 (MDM2) gene, the product of which binds to and inactivates p53, was studied in 60 patients with B-cell chronic lymphocytic leukemia (B-CLL) or non-Hodgkin's lymphoma (B-NHL). Northern blot analysis showed that the level of MDM2 gene expression was low in normal human B-cells, whereas 17 of the patients (28.3%) with B-CLL or NHL had more than 10-fold higher levels of MDM2 gene expression than that observed in normal B cells. Immunohistochemical analysis confirmed MDM2 overexpression at the cellular protein level. MDM2 gene overexpression was found more frequently in patients with the low-grade type of lymphoma (56.5%) than in those with intermediate-/high-grade types (10.8%) (P = .001). Moreover, MDM2 overexpression was found significantly more frequently in patients at advanced clinical stages. Simultaneous analysis of p53 gene mutation showed that three patients had both MDM2 gene overexpression and p53 gene mutation. The results of the present study suggest that MDM2 gene overexpression may play an important role in the tumorigenicity and/or disease progression of CLL and low-grade lymphomas of B-cell origin.

The PTEN/MMAC1 gene at 10q23.3, which has dual specific phosphatase activity, is a novel tumor suppressor gene candidate. Various kinds of tumors have mutations in this gene, including glioblastoma, endometrial carcinoma and prostate cancer. We examined 29 cases of primary non-Hodgkin's lymphoma (NHL) for mutations in the PTEN/MMAC1 gene. One case of diffuse large B cell lymphoma had an 11 bp deletion, but the remaining 28 cases showed no mutations in the genome. Two of these 28 cases showed missense mutations in the PTEN/MMAC1 transcripts, but no alterations in the genomic DNA. These mRNA missense variants are similar to PTEN/MMAC1 transcript aberrations which have been reported in patients with breast cancer. These findings suggest that alterations in the PTEN/MMAC1 gene play a role in the pathogenesis of NHL.

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A rosette plant of Eustoma grandiflorum requires vernalization (exposure to a period of cold temperature) and long-day conditions to promote flowering, while prolonged cold or cool temperatures in post-vernalization periods delay flowering. This study aimed to investigate the effect of growth conditions on flowering regulation in Eustoma. In Arabidopsis, vernalization suppresses a floral repressor gene, FLOWERING LOCUS C (FLC) and upregulates floral promoter genes, such as SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1) and FLOWERING LOCUS T (FT). We identified and characterized the Eustoma homologs of these genes. In contrast to Arabidopsis FLC, Eustoma grandiflorum FLC-like (EgFLCL) expression was upregulated by cold temperature and downregulated by subsequent warm temperature exposure. The expression of Eustoma grandiflorum SOC1-like (EgSOC1L) and FT-like (EgFTL) genes was not significantly induced during vernalization, but their transcripts increased during a warm post-vernalization period in the long days. Vernalized plants grown under cool post-vernalization temperatures exhibited higher EgFLCL expression, lower EgSOC1L and EgFTL expression and flowered later than those grown under warm temperatures. Overexpression of EgFLCL cDNA repressed flowering in transgenic Arabidopsis, whereas overexpression of EgSOC1L or EgFTL cDNA promoted flowering. Our results suggest that flowering regulation by vernalization in Eustoma differs from the paradigm developed for Arabidopsis. EgFLCL is regulated by temperature and may be involved in floral repression during cold and cool seasons. Warm- and long-day conditions following vernalization are required to induce two putative floral promoters, EgSOC1L and EgFTL, effectively.

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ABSTRACT. A polymerase chain reaction (PCR) was developed for the detection of the hemolysin (alpha toxin) gene of Clostridium septicum. The PCR primers were designed from the sequence of the hemolysin gene and synthesized. A DNA fragment of 270 bp was amplified from10 strains of C. septicum, but was not from strains of C. chauvoei, C. perfringens, C. novyi, or C. haemolyticum. When the PCR product was digested with Sau3AI, two DNA fragments of the expected 148 bp and 122 bp were recognized. The lowest detectable threshold of PCR for the hemolysin gene was 3.8 × 10 3 cells/ml. The PCR technique may be useful for rapid detection or identification of C. septicum associated with malignant edema. -KEY WORDS: alpha toxin, Clostridium septicum, PCR.