Mutations of the epigenetics-modifying gene (DNMT3a, TET2, IDH1/2) at diagnosis may induce FLT3-ITD at relapse in de novo acute myeloid leukemia.

Gene mutations were found in acute myeloid leukemia (AML) and their importance has been noted. To clarify the importance and stability of mutations, we examined gene mutations in paired samples at diagnosis and relapse of 34 adult AML patients. Five acquired gene mutations were detected at relapse. Of the 45 gene mutations at diagnosis, 11 of them were lost at relapse. The acquired mutations at relapse were all class I mutations as Fms-like tyrosine kinase 3 (FLT3) and rat sarcoma viral oncogene homolog (RAS) mutations. The disappeared mutations at relapse were 3 of 11 internal tandem duplications of FLT3 (FLT3-ITD) (27.3%), 3 of 3 FLT3 tyrosine kinase domain (FLT3-TKD) (100%), 3 of 13 Nucleophosmin 1 (23.1%) and 2 of 5 CCAAT/enhancer-binding protein-α (40%) mutations. However, epigenetics-modifying gene (DNMT3a, TET2 and IDH1/2) mutations had no change between diagnosis and relapse samples, and may become minimal residual disease marker. The frequency of FLT3-ITD at relapse in patients with DNMT3a mutation at diagnosis is significantly higher than those in patients without them (P=0.001). Moreover, the high frequency of FLT3-ITD at relapse is also seen in AML cases that initially present with any epigenetics-modifying gene mutations (P<0.001). Our results indicate that epigenetics-modifying gene mutations may cause genetic instability and induce FLT3-ITD, leading to resistance to therapy and relapse.

[Surgical management of acute type A aortic dissection with a complaint of disturbance of consciousness; report of a case].

A 50-year-old female was admitted to our hospital with a chief complaint of disturbance of consciousness (DOC). Left-sided hemiparalysis was noted on examination and cerebral infarction was diagnosed with total occlusion of the right common carotid artery revealed by cerebral angiography. Pharmacological thrombolysis (urokinase 720,000 U) was performed. Dissection of the right common carotid artery was noted after successful thrombolytic therapy. Enhanced chest computed tomography (CT) showed the acute type A aortic dissection involving the cerebral artery. Ascending aortic replacement was performed 4 days after the thrombolytic therapy to avoid brain edema and hemorrhagic infarction during cardiopulmonary bypass. The postoperative course was uneventful. In the case of acute type A aortic dissection with DOC, proper indication and optimal timing of the operation may help to improve patient survival.

Gas chromatographic-mass spectrometric determination of hydrophilic compounds in environmental water by solid-phase extraction with activated carbon fiber felt.

Simple gas chromatographic-mass spectrometric determination of hydrophilic organic compounds in environmental water was developed. A cartridge containing activated carbon fiber felt was made by way of trial and was evaluated for solid-phase extraction of the compounds in water. The hydrophilic compounds investigated were acrylamide, N,N-dimethylacetamide, N,N-dimethylformamide, 1,4-dioxane, furfural, furfuryl alcohol, N-nitrosodiethylamine and N-nitrosodimethylamine. Overall recoveries were good (80-100%) from groundwater and river water. The relative standard deviations ranged from 4.5 to 16% for the target compounds. The minimum detectable concentrations were 0.02 to 0.03 microg/l. This method was successfully applied to several river water samples.

Macrophage colony-stimulating factor induces the growth and differentiation of normal pregnancy human cytotrophoblast cells and hydatidiform moles but does not induce the growth and differentiation of choriocarcinoma cells.

In the present study, we examined whether or not macrophage colony-stimulating factor (M-CSF; CSF-1) is involved in the growth and differentiation of human chorionic, hydatidiform mole and choriocarcinoma cells. M-CSF promotes the growth of early gestation chorionic cells, hydatidiform mole cells, and a human term placenta cell line (tPA30-1). However, the growth of choriocarcinoma cells, BeWo, Jar, Jeg-3, and NUC-1, was not influenced at all by M-CSF. M-CSF promoted the secretion of human chorionic gonadotropin (hCG) and human placental lactogen (hPL), which are secreted from differentiated trophoblast, from early gestation chorionic cells and from hydatidiform mole cells. However, the secretion of hCG and hPL from choriocarcinoma cells was not affected by M-CSF. When M-CSF localization was examined by immunohistochemical staining, M-CSF was detected in chorionic and hydatidiform mole cells, but was absent in choriocarcinoma cells. These results suggest that the growth and differentiation of normal chorionic and hydatidiform mole cells are M-CSF-dependent, while the growth and differentiation of choriocarcinoma cells are not.

Transacting activities of the E7 genes of several types of human papillomavirus.

In accordance with previous reports by others, the E7 gene of human papillomavirus (HPV) 16, considered to be etiologically associated with cervical cancer, transactivated the E2 promoter of adenovirus. This promoter, however, was equally stimulated by the E7 gene of HPV1, a skin type HPV never associated with human malignancy. A variety of promoters were tested to see the effect of the E7 genes of low-risk and high-risk type HPVs. The result indicated that there was no obvious relationship in the levels of transactivation or transrepression by the E7 gene between low-risk and high-risk types. It has been suggested that the binding of the E7 protein to the product of retinoblastoma susceptibility gene (pRb) is the underlying mechanism by which the E7 protein transactivates the E2 promoter. Therefore, the association of the E7 protein and pRb alone did not seem to fully explain the mechanism by which this protein participates in the activation of transcription and induction of human malignancies.

Use of universal and type-specific primers in the polymerase chain reaction for the detection and typing of genital human papillomaviruses.

By using polymerase chain reaction (PCR), we have developed a system for type-specific as well as universal detection of genital human papillomaviruses (HPVs). Primers and probes for specific detection of HPV-16, -18 and -33 were synthesized from the E7 open reading frame (ORF). They were capable of detecting corresponding HPV types with high specificity and sensitivity. Primers for detection of a broad spectrum of HPV (universal primers) were synthesized from the L1 ORF. The universal primers were shown to be capable of amplifying HPV-6b, -11, -16, -18, -33, -52b and -58. The system was applied to various cervical tissue specimens from Japanese patients. They consisted of 26 normal specimens, 18 from cervical dysplasias and 29 from cervical carcinomas. HPV was detected in none of the normal specimens. On the other hand, many of the specimens from cervical dysplasias and carcinomas were found to be positive for HPV, especially HPV-16. Except for one, all the specimens which were positive with the type-specific PCRs were also positive with the universal PCR. Furthermore, substantial numbers of specimens were found to be positive only with the universal PCR. Cloning and sequencing of DNA segments amplified by the universal primers were undertaken to characterize some of the unknown HPVs. Our PCR system may thus be useful for the specific detection of the three major types of oncogenic HPVs and also for the detection of a broad spectrum of HPVs including possibly novel HPV types.

Loss of responsiveness of an AP1-related factor, PEBP1, to 12-O-tetradecanoylphorbol-13-acetate after transformation of NIH 3T3 cells by the Ha-ras oncogene.

The function of the A element (nucleotides 5107 to 5130) of the polyomavirus enhancer is augumented in NIH 3T3 cells by a tumor-promoting phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA). One of its targets is an AP1 consensus sequence motif recognized by a nuclear factor, PEBP1. In Ha-ras-transformed NIH 3T3 cells, however, A element function was not enhanced by TPA treatment, and at the same time PEBP1 was not detected in the nuclear extract by a mobility shift assay. PEBP1 was not detected in either the extract from NIH 3T3 cells treated in vivo with a protein kinase inhibitor, staurosporine, or the extract from NIH 3T3 cells after treatment in vitro with phosphatase. These results suggest that PEBP1 is required to be properly phosphorylated for DNA binding and that it is underphosphorylated, possibly due to the downregulation of protein kinase C in Ha-ras-transformed cells. In addition, we observed that PEBP2, which bound to the A element adjacent to PEBP1, was converted to apparently related PEBP3 when conditions favored underphosphorylation.

[A case report of advanced ovarian cancer with chronic renal dysfunction treated by CDDP i.p].

Reported in this paper is a case of a 56-year-old female patient who was diagnosed as having an ovarian cancer (stage T 4) with a renal dysfunction (CCr 30.5 ml/min). We instilled CDDP intra-peritoneally (CDDP-ip), taking into consideration the peritoneal clearance (CLp.). The administration of sodium thiosulphate also was combined. The low CLp. of this case (299.2 ml/h) made it possible to instill a high dose in the peritoneal cavity and a low dose in the plasma. This method provided enough of anti-tumor effect and kept the renal function safe. We believe that the CLp. is the most important factor in determining the effect and side effects of CDDp-ip.

[A case report of an advanced ovarian cancer (stage T4) treated with a total amount of 1815 mg of CDDP].

A case is reported of a 34-year-old female who complained of an adult head-sized tumor in the pelvis and was treated with a high dose of CDDP. On the first look operation, it was found impossible to perform reduction surgery because of severe adhesion and peritonitis carcinomatosa. To introduce a high concentration of an anti-cancer agent into the tumor tissue, we administered CDDP intravenously, intraperitoneally and intra-arterially to the total amount of 1815 mg. After this high dose combination chemotherapy, a second look operation was performed and the tumor was seen to be reduced. The patient is now in remission and is being followed up as an out-patient. The clinical course of this case had decided for us which route would be the most effective and would have the least side effect, also, how much of a dosage can be administered as a maximum adoptive dose of an anti-cancer agent, and what kind of a second-line chemotherapy is effective against resistant cancer cells.

[Göttingen miniature swine as a model for diet-induced atherosclerosis].

Twenty-four Göttingen Miniature Swine/csk, in order to evaluate their potential usefulness as a model for experimental atherosclerosis studies, were fed diets of three types, a high-fat plus high cholesterol diet, a high-fat diet, and a commercial diet. Each group consisted of 4 males and 4 females. Swine fed the experimental diet were investigated by gross, microscopic and serum biochemical examination on the 1st, 3rd, 6th and 9th month after start of experimentation. Lesions of atherosclerosis were observed in the high-fat plus high-cholesterol diet group. After a month on the experimental diet, intimal thickening was detected in the abdominal aorta just above the origin of internal iliac artery, left coronary artery and ascending aorta by microscopic examination. Thereafter, on the 9th month after the start, there was more extensive and severe atherosclerosis. These lesions were classified into two types by the difference in the histologic architecture of arterial wall. One was fatty streaks that were in thoracic aorta belonging to the elastic type and the other was fibrous plaques that were in abdominal aorta and iliac artery and so on, belong to the transitional or muscular type. High-fat plus high-cholesterol diet feeding led to elevated serum cholesterol and beta-lipoprotein levels, and had an effect on several kinds of metabolism. All of the swine fed high-fat or commercial diet had little gross, microscopic lesions and had no change in serum cholesterol and beta-lipoprotein levels. Hypercholesterolemia and hyper-beta-lipoproteinemia had a close relation to the development and acceleration of atherosclerosis. It was possible to show that the diet induced atherosclerosis was similar in quality to that observed in humans, and that the Göttingen miniature swine was a suitable animal for the study of atherosclerosis.