Wortmannin enhances CPP32-like activity during neuronal differentiation of P19 embryonal carcinoma cells induced by retinoic acid

P19 EC cells undergoes apoptosis during neuronal differentiation induced by retinoic acid. Two CPP32-like proteases, CPP32 and Mch-3, are expressed in untreated and retinoic acid-treated P19 EC cells. CPP32-like activity is remarkably increased in apoptosis during neuronal differentiation of P19 EC cells. Inhibition of CPP32-like proteases prevents apoptosis, suggesting that activation of CPP32-like proteases play central roles in the apoptosis during neuronal differentiation of P19 EC cells. Wortmannin, PI-3K inhibitor, enhances the CPP32-like activity of the retinoic acid-treated P19 EC cells. PI-3K may be involved in the apoptosis during neuronal differentiation as negative regulator.     

Cathepsin E expression by normal and premalignant cervical epithelium

We have investigated the expression of the aspartic proteinase cathepsin E and HLA-DR and the presence of HPV16 in normal squamous epithelium (n = 8) and low-grade (n = 21) and high-grade (n = 14) intraepithelial squamous lesions of the uterine cervix. Immunohistochemistry of cervical biopsies revealed that up-regulation of cathepsin E expression was related to increasing severity of the cervical intraepithelial neoplasia (CIN). Up-regulation of protein was associated with increased message as assessed by in situ hybridization. Langerhans cells and the majority of koilocytes did not express detectable cathepsin E levels. Although there was also an up-regulation of HLA-DR expression by cervical keratinocytes in cervical intraepithelial neoplasia lesions, as determined by immunohistochemistry, no significant correlation was found between HLA-DR and cathepsin E expression in these lesions; neither was expression of cathepsin E correlated to the presence of HPV16, detected by polymerase chain reaction. The expression of cathepsin E, an aspartic proteinase that is reported to play a role in antigen processing for presentation by class II major histocompatibility complex molecules, is associated with cellular dedifferentiation in cervical intraepithelial neoplasia.     

Screening mixtures for biological activity by NMR

Development of the new drugs often involves the screening of compound libraries for biological activity. Currently, the biologically active component can only be identified if either a pure compound is being tested or if the components of a mixture are spatially separated, for example, on beads. Here, we present an NMR technique based on the transferred nuclear Overhauser effect (transfer NOE) that allows identification and structural characterization of biologically active molecules from a mixture. As an example we demonstrate that from mixtures of oligosaccharides only alpha-L-Fuc-(1-->6)-beta-D-GlcNAc-OMe binds to Aleuria aurantia agglutinin. The sign of transferred NOEs is opposite to NOEs of small molecules that do not bind to the protein and, thus, an unequivocal identification of molecules with binding activity is possible. Normally, the selection of bound ligands is further facilitated in that the absolute intensity of transfer NOEs is much greater than that of NOEs of non-binding molecules. In addition, transfer NOEs provide information on the three-dimensional structure of the ligands in the bound state. Therefore, measuring transfer NOEs of mixtures of small molecules in the presence of large molecules, like proteins, should significantly enhance the options for screening mixtures of compounds for biological activity.     

The effects of betamethasone derivatives on endotoxin-induced uveitis in guinea pigs

OBJECTIVE AND DESIGN: We reported previously that the betamethasone derivative betamethasone dipropionate behaves as an anti-glucocorticoid in rat endotoxin-induced uveitis (EIU). In the present study, we produced EIU in guinea pigs and investigated the effects of betamethasone dipropionate on the EIU. MATERIAL: Male Hartley guinea pigs were used. TREATMENT: Glucocorticoids were instilled into the eye. METHOD: To elicit EIU, lipopolysaccharide (LPS) was injected into the anterior chamber of the eye. Cell numbers in the aqueous humor after LPS injection were determined by flow cytometry. Prostaglandin E2 (PGE2) production after LPS injection into the anterior chamber was also examined. RESULTS: Intracameral injection of LPS (1 microgram/eye) induced cell infiltration into the anterior chamber and PGE2 production. Betamethasone dipropionate inhibited cell infiltration and PGE2 production more strongly than betamethasone. These results suggest that betamethasone dipropionate is a potent glucocorticoid in guinea pigs. CONCLUSIONS: Structure-activity relationships of glucocorticoids in the guinea pig EIL model may differ from those in the rat EIU model.     

Clinical oncology nurses' perceptions of research

Within the realm of oncology nursing, research has been an integral part in its development as a specialty practice. Yet despite the growing volume of published nursing research studies, little is known about how nurses working in oncology care settings perceive research. Therefore, the purposes of this study were to examine clinical oncology nurses' perceptions of research and to determine factors influencing their perceptions. Two hundred and eighty-three registered nurses providing cancer care to patients in 40 health care agencies across northern Ontario participated in the survey. Data were collected using a questionnaire developed by Alcock and colleagues (1990) which addressed nurses' perceived value of research, their role, interest and experience in research as well as the research climate of the agency. The findings showed that respondents valued nursing research and perceived a research role for staff nurses. However, the respondents did not perceive strong administrative or collegial support for nurses' involvement in research activities. In addition, the study results indicated that the clinical oncology nurses' perceptions of research were influenced by educational preparation.     

Automatic tuning of decentralized PID controllers for MIMO processes

An automatic tuning algorithm for decentralized PID control in multiple-input multiple-output (MIMO) plants is presented. This algorithm generalizes the authors' recent auto-tuner for two-input two-output systems to any number of inputs and outputs. The algorithm consists of two stages. In the first, the desired critical point, which consists of the critical gains of all the loops and a critical frequency, is identified. The auto-tuner identifies the desired critical point with almost no a priori information about the process. During the identification phase all controllers are replaced by relays, thus generating limit cycles with the same period in all loops. It is shown that each limit cycle corresponds to a single critical point of the process. By varying the relays parameters different points can be determined. The auto-tuner contains a procedure which converges rapidly to the desired critical point while maintaining the amplitudes of the process variables as well as of the manipulated variables within prespecified ranges. In the second stage, the data of the desired critical point is used to tune the PID controllers by the Ziegler-Nichols rules or their modifications. This paper focuses on the first stage. The steady-state process gains, which are required for the appropriate choice of the desired critical point, are determined by the auto-tuner in closed-loop fashion simultaneously with the identification of the critical point. The identification of the process gains is achieved at no extra plant time. Based upon a large number of simulated cases, the proposed auto-tuner seems to be efficient and robust. The paper discusses the underlying principles of the auto-tuner and its properties and capabilities are demonstrated via examples.     

Evaluating the impact of CASE: an empirical comparison of retrospective and cross-sectional survey approaches

The purpose of this paper is to evaluate two methods of assessing the productivity and quality impact of Computer Aided Software Engineering (CASE) and Fourth Generation Language (4GL) technologies: (1) by the retrospective method; and (2) the cross-sectional method. Both methods involve the use of questionnaire surveys. Developers' perceptions depend on the context in which they are expressed and this includes expectations about the effectiveness of a given software product. Consequently, it is generally not reliable to base inferences about the relative merits of CASE and 4GLs on a cross-sectional comparison of two separate samples of users. The retrospective method that requires each respondent to directly compare different products is shown to be more reliable. However, there may be scope to employ cross-sectional comparisons of the findings from different samples where both sets of respondents use the same reference point for their judgements, and where numerical rather than verbal rating scales are used to measure perceptions.     

Preferential exclusion of sucrose from recombinant interleukin-1 receptor antagonist: role in restricted conformational mobility and compaction of native state

Understanding the mechanism for sucrose-induced protein stabilization is important in many diverse fields, ranging from biochemistry and environmental physiology to pharmaceutical science. Timasheff and Lee [Lee, J. C. & Timasheff, S. N. (1981) J. Biol. Chem. 256, 7193-7201] have established that thermodynamic stabilization of proteins by sucrose is due to preferential exclusion of the sugar from the protein's surface, which increases protein chemical potential. The current study measures the preferential exclusion of 1 M sucrose from a protein drug, recombinant interleukin 1 receptor antagonist (rhIL-1ra). It is proposed that the degree of preferential exclusion and increase in chemical potential are directly proportional to the protein surface area and that, hence, the system will favor the protein state with the smallest surface area. This mechanism explains the observed sucrose-induced restriction of rhIL-1ra conformational fluctuations, which were studied by hydrogen-deuterium exchange and cysteine reactivity measurements. Furthermore, infrared spectroscopy of rhlL-1ra suggested that a more ordered native conformation is induced by sucrose. Electron paramagnetic resonance spectroscopy demonstrated that in the presence of sucrose, spin-labeled cysteine 116 becomes more buried in the protein's interior and that the hydrodynamic diameter of the protein is reduced. The preferential exclusion of sucrose from the protein and the resulting shift in the equilibrium between protein states toward the most compact conformation account for sucrose-induced effects on rhIL-1ra.