Amino acid residues contributing to stabilization of Fusarium heterosporum lipase

Fusarium heterosporum lipase is composed of an N-terminal large peptide of 275 amino acids and a C-terminal peptide of 26 amino acids. The thermostability of the lipase was remarkably decreased by cleavage of the C-terminal peptide. Hence, we attempted to specify the amino acids in the C-terminal peptide that are responsible for the stabilization of the lipase. Replacement of Asp293 with Ala, Asn, and Lys caused a significant decrease in thermostability, but its mutation to Glu did not decrease the stability significantly. These findings showed that the lipase with the C-terminal peptide was stabilized by an ionic bond between the negative charge of Asp293 and positive charge of an amino acid of the N-terminal large peptide. The thermostability of the lipase gradually decreased with increasing deletion size from the C-terminus, and a 13-amino acid deletion decreased the stability to the level of the lipase not having the C-terminal peptide. These results suggested that the 13-amino acid region from the C-terminus participated in the lipase stability. In addition, the lipase production correlated well with the lipase stability, showing that the C-terminal peptide also influenced the lipase productivity.     

Marked eosinophilia in interleukin-5 transgenic mice fails to prevent Trichinella spiralis infection

In order to study the role of eosinophils in the host defense against Trichinella spiralis infection, worm recovery after infection with T. spiralis was compared between interleukin-5 transgenic (IL-5 Tg) mice with a constant high level of peripheral eosinophils and nontransgenic C3H/HeN mice. No significant difference in the recovery of muscle larvae or adult worms in the small intestine, fecundity of female adult worms, or infectivity of newborn larvae was observed between nonimmunized C3H/HeN and IL-5 Tg mice or C3H/HeN and IL-5 Tg mice immunized with somatic antigen of T. spiralis. However, a significant difference was observed in the fecundity of female adult worms and recovery of muscle larvae between nonimmunized and immunized IL-5 Tg mice or C3H/HeN mice. These results demonstrate that having more eosinophils does not improve immunity against the various aspects of T. spiralis infection.     

Plasma factor triggering alternative complement pathway activation by liposomes

Several plasma components, such as complement (C) components, play a role in the clearance of liposomes from the circulation. The interactions between liposomes and the C system were investigated in this study. Multilamellar vesicle (MLV) liposomes, which were damaged by activation of the complement, became susceptible depending on the density of cetylmannoside (Man) on the liposome membrane, and activation proceeded through the alternative C pathway as observed for liposomes without Man (PC-MLV) (K. Funato et al., Biochim. Biophys. Acta 1103:198-204, 1992). In addition, the capacity of Man-modified liposomes (Man-MLV) to activate the alternative C pathway was abolished by preadsorption of plasma with Man-MLV but not with PC-MLV. The results suggest that a specific plasma factor adsorbed with Man-MLV was responsible for the augmentation of the C activation and, further, that the rapid clearance of Man-MLV from the circulation is caused by both enhanced C-mediated liposome permeability and enhanced C-mediated phagocytosis of liposomes.     

A late phase II study of CPT-11 (irinotecan) in advanced breast cancer. CPT-11 Study Group on Breast Cancer

A late phase II study of CPT-11 for advanced breast cancer was conducted at 27 institutions. Seventy-nine patients were enrolled, 75 were eligible for the study, and 65 were evaluable for efficacy. One complete response and 14 partial responses were obtained, and the response rate was 23%. The response rate of patients with prior endocrine therapy and prior chemotherapy including adriamycin or other anthracycline drugs was 27% (11/41) and 26% (12/46), respectively. The response rate for patients with estrogen receptor-negative tumors and premenopausal patients was 32% (6/19) and 27% (4/15), respectively. Responses were observed not only for soft tissue lesions such as lymph nodes (5/17), but also for distant metastases in the lungs (8/28) and bone (1/18). The major adverse reactions were myelosuppression and gastrointestinal symptoms. The incidence of Grade 2 or higher leukopenia, anemia, nausea/vomiting, anorexia, diarrhea and alopecia was 68%, 31%, 67%, 59%, 37%, and 30%, respectively. These results suggested that CPT-11 was a promising drug for advanced breast cancer.     

AT1 receptor-mediated stimulation by angiotensin II of rat aortic fibronectin gene expression in vivo

Fibronectin plays an important role in various vascular diseases. A subpressor (200 ng kg-1 min-1) or pressor (1000 ng kg-1 min-1) dose of angiotensin II was continuously infused into rats by osmotic minipump for various times, to investigate the effects on aortic fibronectin gene expression. In rats infused with a subpressor dose of angiotensin II in which blood pressure was normal for 3 days, aortic fibronectin mRNA levels started to increase by 1.4 fold at 12 h and reached the maximal levels (increased by 3.1 fold) at 3 days. Treatment with TCV-116 (3 mg kg-1 day-1), a non-peptide selective AT1 receptor antagonist, completely inhibited the angiotensin II-induced increase in aortic fibronectin mRNA, while hydralazine (10 mg kg-1 day-1) did not block this effect. Similar results were also obtained for a pressor dose of angiotensin II. Thus, angiotensin II directly stimulates aortic fibronectin gene expression in vivo, which is mediated by the AT1 receptor but not by blood pressure.     

Determination of sulphur heterocycles in selected synfuels

A fractionation method for isolating the polycyclic aromatic sulphur heterocycles from complex mixtures of polycyclic aromatic compounds was evaluated with respect to recovery for selected standard compounds. It was found that recovery varied from 0 to 70% depending on the structure of the compound. Although quantitative recovery for all components was not obtained, isolation of the sulphur heterocycle fractions has allowed the identification of many new sulphur compounds previously not identified in coal liquids and shale oils. Capillary column gas chromatography with flame photometric detection and mass spectrometry were used for the identification of individual sulphur heterocycles.     

Significance of circulating hepatocyte growth factor level as a prognostic indicator in primary breast cancer

OBJECTIVE: The purpose of the study was to develop quantitative and qualitative criteria for diagnosing fatty liver on contrast-enhanced helical CT. SUBJECTS AND METHODS: Differential liver-spleen attenuation was evaluated between 80 and 120 sec after injection in 76 patients who underwent contrast-enhanced helical CT. Unenhanced CT images had earlier established fatty liver when the liver minus spleen attenuation difference was less than or equal to -10 H (n = 18). Four observers who had not seen the unenhanced images used contrast-enhanced CT images to assess the presence of fatty liver on a five-point Likert scale, the presence of geographic areas spared from fatty infiltration, and the relative liver-spleen attenuation. The diagnostic accuracies of various imaging criteria were compared using McNemar's chi-square test (for sensitivity and specificity) and analysis of receiver operating characteristic curves. RESULTS: Sensitivity, specificity, and receiver operating characteristic curve areas for observers' qualitative judgments were 54%, 95%, and .91, respectively; for quantitative differential liver-spleen attenuation (80-100 sec; -20.5 H discriminatory value), the values were 86%, 87%, and .94, respectively; and for quantitative differential liver-spleen attenuation (101-120 sec; -18.5 H discriminatory value), the values were 93%, 93%, and .98, respectively. Differential liver-spleen attenuation was time-dependent; overlap was noted between healthy subjects and patients with fatty liver. Qualitatively, geographic sparing was highly specific (94%) for fatty liver, whereas liver attenuation greater than or equal to spleen attenuation excluded fatty liver in all but one case. CONCLUSION: Although quantitative and qualitative criteria for diagnosing fatty liver on helical CT can be determined, they are protocol-specific. Limited unenhanced hepatic CT remains the optimal technique for detection of fatty infiltration of the liver.     

Sensory Modeling of Coffee with a Fuzzy Neural Network

ABSTRACT: Models were constructed to predict sensory evaluation scores from the blending ratio of coffee beans. Twenty-two blended coffees were prepared from 3 representative beans and were evaluated with respect to 10 sensory attributes by 5 coffee cup-tasters and by models constructed using the response surface method (RSM), multiple regression analysis (MRA), and a fuzzy neural network (FNN). The RSM and MRA models showed good correlations for some sensory attributes, but lacked sufficient overall accuracy. The FNN model exhibited high correlations for all attributes, clearly demonstrated the relationships between blending ratio and flavor characteristics, and was accurate enough for practical use. FNN, thus, constitutes a powerful tool for accelerating product development.