Biochemical identification of transmembrane segments of the Ca(2+)-ATPase of sarcoplasmic reticulum

The transmembrane segments of sarcoplasmic reticulum Ca(2+)-ATPase were determined by trypsinization of cytoplasmic side-out intact sarcoplasmic reticulum vesicles. The membrane portion of tryptic digest comprising the transmembrane fragments, joined by the intravesicular segments, was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis after labeling with fluorescein 5-maleimide in the presence of sodium dodecyl sulfate. In this way, seven fluorescent bands of tryptic fragments below 11 kDa were observed which were derived from 4 pairs of membrane spanning segments and one hydrophobic sequence at the C-terminal end. Two peptides of 10.8 and 10.6 kDa had the identical N-terminal sequence beginning at Glu826, representing the transmembrane segments M7 and M8 and their connecting loop. A band at 8.1 kDa contained one peptide beginning at Tyr36 (M1/loop/M2). A 7.7-kDa peptide starting at Leu253 (M3/loop/M4) and a 7.3-kDa peptide beginning at Ala752 (M5/loop/M6) were also observed. A band at 6.7 kDa contained two peptides, one beginning at Ser48 (M1/loop/M2) and another beginning at Tyr763 (M5/loop/M6). In addition, a 4-kDa peptide beginning at Met925 was observed. The size of this peptide did not allow for a complete pair of transmembrane segments, but this peptide could have been derived from trypsinolysis between the last pair of membrane spanning segments. These data therefore provide biochemical evidence for at least 8 transmembrane segments and perhaps two more at the C-terminal end of the enzyme.     

Peptide YY inhibition of rat gastric enterochromaffin-like cell function

The present study determined tumorigenicity, tumor classification and DNA damage induced in infant mice by benzo[a]pyrene (B[a]P) or Manufactured Gas Plant (MGP) residues after a single exposure. Male and female B6C3F1 mice were exposed to B[a]P or MGP residue from a single environmental site (MGP-4) and males were also exposed to MGP residue composite from seven different sites (MGP-M7). At 26, 39 and 52 weeks after exposure tumorigenesis was assessed in lung, forestomach and liver. Formation and persistence of DNA adducts were quantified by 32P-postlabeling. Exposure of males to B[a]P induced liver tumors in a dose and time dependent manner. MGP induced more advanced tumors than B[a]P. Only a single liver tumor was found in MGP-4 treated females. No forestomach and few pulmonary adenomas were induced in males or females. MGP-4, MGP-M7 or B[a]P induced DNA adducts in males and females. Adducts in liver, lung and forestomach peaked on different days and decreased at different rates. At 24 h post-exposure, no significant differences in initial DNA adduct levels occurred in males and females exposed to MGP-4 or B[a]P. Lack of DNA damage (adducted DNA) did not account for non-responsiveness of lung and forestomach in B6C3F1 genders as well as in liver in females. MGP tumorigenicity could not be accounted for solely by B[a]P content nor did it reflect additivity of B[a]P and other carcinogenic polycyclic aromatic hydrocarbons (PAHs) in MGP. Synergy among MGP-PAHs, presence of unidentified carcinogens and/or promoters in MGP may account for MGP potency. The B6C3F1 infant male model is a convenient and rapid assay for assessing MGP liver tumorigenicity and potency.     

Penile responses of rats (Rattus norvegicus) in extended ex copula tests.

Conducted prolonged tests of male Long-Evans rats' penile responses outside the context of copulation (ex copula) to facilitate analysis of these penile components of copulatory behavior. Penile erections and flips were evoked for 1 hr following penile sheath retraction (SR), and Ss were retested for 30 min after a rest interval of 5 min to 4 hr. The number of penile responses declined sharply over the course of 1 hr. Following a 5-min rest, there was little likelihood of additional responses. Within 2 hrs, some measures of penile response potential approached their original high asymptotes: other measures were still depressed after the 4-hr rest. In control tests, the penile sheath was unretracted (SU) during the 1st 60 min, and hence few or no penile responses occurred. This condition caused no reduction in the number of erections and flips in the subsequent 30-min SR test; hence, the reduction in response to potential during and after SR tests was due to the responses displayed, not to the conditions of restraint. Ejaculation occurred frequently in the SU condition but rarely in SR tests. These results suggest that SR may normally inhibit ejaculation in ex copula tests and during copulation. (41 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

PEAK-INSENSITIVE NON-PARAMETRIC SPECTRUM ESTIMATION

Abstract. We study the problem of non-parametric spectrum estimation of a stationary time series that might contain periodic components. In this case the periodogram ordinates have a significant amplitude at frequencies near the frequencies of the periodic components. These can be regarded as outliers in an asymptotically exponential sample. We develop a non-parametric estimator for the spectral density that is insensitive to these outliers in the frequency domain. This is done by robustifying the usual kernel estimator (smoothed periodogram) by means of M-estimation in the frequency domain. We propose to use data-tapered periodograms, which yield a drastic improvement of the procedure, typically for the contaminated situation. This is both shown theoretically and supported by means of simulation. We show consistency of the resulting estimator in the general case, and asymptotic normality in the special case of a Gaussian time series, whether contamination is present or not. Finally we illustrate the finite sample performance of the estimating procedure by some simulation results and by application to the Canadian lynx trappings data.     

Performance modeling and analysis of message-oriented event-driven systems

Message-oriented event-driven systems are becoming increasingly ubiquitous in many industry domains including telecommunications, transportation and supply chain management. Applications in these areas typically have stringent requirements for performance and scalability. To guarantee adequate quality-of-service, systems must be subjected to a rigorous performance and scalability analysis before they are put into production. In this paper, we present a comprehensive modeling methodology for message-oriented event-driven systems in the context of a case study of a representative application in the supply chain management domain. The methodology, which is based on queueing Petri nets, provides a basis for performance analysis and capacity planning. We study a deployment of the SPECjms2007 standard benchmark on a leading commercial middleware platform. A detailed system model is built in a step-by-step fashion and then used to predict the system performance under various workload and configuration scenarios. After the case study, we present a set of generic performance modeling patterns that can be used as building blocks when modeling message-oriented event-driven systems. The results demonstrate the effectiveness, practicality and accuracy of the proposed modeling and prediction approach.     

A structural linearization principle for processes

In [11], an induction principle for processes was given which allows one to apply model-checking techniques to parameterized families of processes. A limitation of the induction principle is that it does not apply to the case in which one process depends directly upon a parameterized number of processes, which grows without bound. This would seem to preclude its application to families ofN processes interconnected in a star topology. Nonetheless, we show that if the dependency can be computed incrementally, then the direct dependency upon the parameterized number of processes may be re-expressed recursively in terms of a linear cascade of processes, yielding in effect a linearization of the inter-process dependencies and allowing the induction principle to apply.A previous version of this paper appears in the Proceedings of CAV 1993 (LNCS 697).