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71.
Encapsulation of soluble protein antigens in liposomes was previously shown to result in processing of antigen via the major histocompatibility complex class I pathway, as evidenced by costaining of the trans-Golgi region of murine bone marrow-derived macrophages (BMs) by fluorophore-labeled liposomal antigen and by a trans-Golgi-specific fluorescent lipid. Evidence is presented here that free or liposome-encapsulated RTS,S, a particulate malaria antigen consisting of hepatitis B particles coexpressed with epitopes from the Plasmodium falciparum circumsporozoite protein, also was localized in the trans-Golgi after incubation with BMs, suggesting processing by the class I pathway. An in vivo cytotoxic T-lymphocyte (CTL) response was detected, however, only after immunization with RTS,S encapsulated in liposomes containing lipid A and not after immunization with free RTS,S or with RTS,S encapsulated in liposomes lacking lipid A. Therefore, intracellular delivery of antigen containing CTL epitopes to the Golgi of BMs does not necessarily result in a CTL response in vivo unless an additional adjuvant, such as liposomes containing lipid A, is utilized. Encapsulation of RTS,S in liposomes containing monophosphoryl lipid A (MPL) resulted in a dose-dependent enhancement of the NANP-specific immunoglobulin G (IgG) antibody response compared to that of free RTS,S. The IgG1 and IgG2a subclasses predominated after immunization with RTS,S encapsulated in liposomes containing MPL. These results demonstrate that encapsulation of a lipid-containing particulate antigen, such as RTS, S, in liposomes containing lipid A can enhance both humoral and cellular immune responses.  相似文献   
72.
The recent trends in portable computing technologies have established the need for energy efficient design strategies. To achieve minimum energy design goals, system designers need a technique to accurately model the energy consumption of their design alternatives without performing a full physical design and full-circuit simulation. This paper presents and compares five approaches for modeling the energy consumption of CMOS circuits. These five modeling approaches have been chosen to represent the various levels of model complexity and accuracy found in the current literature. These modeling approaches are applied to the energy consumption of SRAM's to provide examples of their use and to allow for the comparison of their modeling qualities. It was found that a mixed characterization model-using a CV2 prediction for digital subsections and fitted simulation results for the analog subsections-is satisfactory (within ±1 process variation) for predicting the absolute energy consumed per cycle. This same model is also very good (within 2%) for predicting an optimum organization for the internal structures of the SRAM. Several common architectures and circuit designs for SRAM's are analyzed with these models. This analysis shows that global, rather than local improvements, produce the largest energy savings  相似文献   
73.
Carbamoyl-phosphate synthetase (CPSase) consists of a 120-kDa synthetase domain (CPS) that makes carbamoyl phosphate from ATP, bicarbonate, and ammonia usually produced by a separate glutaminase domain. CPS is composed of two subdomains, CPS.A and CPS.B. Although CPS.A and CPS.B have specialized functions in intact CPSase, the separately cloned subdomains can catalyze carbamoyl phosphate synthesis. This report describes the construction of a 58-kDa chimeric CPSase composed of Escherichia coli CPS.A catalytic subdomains and the mammalian regulatory subdomain. The catalytic parameters are similar to those of the E. coli enzyme, but the activity is regulated by the mammalian effectors and protein kinase A phosphorylation. The chimera has a single site that binds phosphoribosyl 5'-pyrophosphate (PRPP) with a dissociation constant of 25 microM. The dissociation constant for UTP of 0.23 mM was inferred from its effect on PRPP binding. Thus, the regulatory subdomain is an exchangeable ligand binding module that can control both CPS.A and CPS.B domains, and the pathway for allosteric signal transmission is identical in E. coli and mammalian CPSase. A deletion mutant that truncates the polypeptide within a postulated regulatory sequence is as active as the parent chimera but is insensitive to effectors. PRPP and UTP bind to the mutant, suggesting that the carboxyl half of the subdomain is essential for transmitting the allosteric signal but not for ligand binding.  相似文献   
74.
A psychrotrophic yeast, Rhodotorula glutinis KUJ 2731, isolated from soil, effectively produced an extracellular endo-beta-glucanase (EC 3.2.1.4). The enzyme was monomeric, and the molecular mass was about 40,000 Da. The N-terminal amino acid sequence was H-Ser-Leu-Pro- Lys-Leu-Gly-Gly-Val-Asp-Leu-Ala-Gly-Leu-Asp-Ile-Gly-Lys-Asp-Lys-Asn-. alpha-Helix content was calculated to be about 32.6%. The isoelectric point was 8.57. The activation energy was 20.9 kJ/mol, which was much smaller than that of mesophilic enzymes. The enzyme was active at temperatures from 0 to 70 degrees C, with a highest initial velocity at 50 degrees C similar to other psychrotrophic enzymes. The enzyme was inhibited by Hg2+. The enzyme catalyzed hydrolysis of carboxymethyl cellulose with an apparent K(m) of 1.1% and Vmax of 556 mumol/min/mg. Products from the enzymatic hydrolysis of carboxymethyl cellulose by the enzyme were glucose, cellobiose, and cellotriose. The enzyme also catalyzed the transglycosylation of p-nitrophenyl-beta-cellotrioside to cellotetraose.  相似文献   
75.
Criteria for detection of chromosome aberrations by Comparative Genomic Hybridization (CGH) are not standardized and improvement of this part of the analysis is of paramount importance to the applicability of the technique. The aim of this work was to suggest CGH detection criteria that increase the specificity and sensitivity and at the same time include chromosome regions previously excluded from CGH analysis. We analyzed 33 hybridizations with normal DNA and modified our CGH software in order to use a selection of these normal analyses as a model for interpretation of analyses of unknown samples. This approach was successfully tested on 14 samples with known aberrations.  相似文献   
76.
77.
Endotoxaemia is a leading cause of death among horses. Thrombocytopenia is a common finding in clinical and experimentally-induced cases of endotoxaemia and can lead to coagulopathies, including disseminated intravascular coagulopathy which is usually fatal. In this study it was shown that endotoxin (3 ng ml-1 to 25 micrograms ml-1) can aggregate equine platelets in heparinised whole blood in vitro. The endotoxin-induced aggregation (EIA) was shown to be dependent on the presence of leucocytes in the blood and did not occur when detoxified endotoxin was used, suggesting that lipid A was necessary for the response. Aspirin (1 mmol litre-1) had no effect on EIA whereas apyrase (40 micrograms ml-1) completely abolished it and CV3988 (3 to 30 mumol litre-1) (a competitive antagonist of platelet-activating factor) inhibited the response in a concentration-dependent manner. It is concluded that endotoxin activates equine platelets at low concentrations through an indirect mechanism that involves calcium, leucocytes, adenine nucleotides and platelet-activating factor.  相似文献   
78.
Experimental studies have been performed on ceramic matrix composite (CMC) I-sections, which typify joint designs for CMC components. Axial loads and moments have been applied to activate delamination mechanisms. The maximum load bearing capacity has large variability, governed by the severity of manufacturing flaws located in the transition region of the I-section. This variability leads to an unsatisfactory design situation. Delaminations that form from these flaws arrest and behave in a stable manner, subject to a remanent load bearing capacity. This remanent capacity has minimal variability. Hence, design based on the remanent load would be robust. An expression for this design criterion is presented.  相似文献   
79.
A laboratory study was undertaken to assess the relationship between the maximal voluntary isometric strength (MVIS) of a group of Hong Kong Cantonese males and recommended limiting values for lifting proposed in the Work Practices Guide for Manual Lifting (NIOSH, 1981). Forty-one male subjects were required to apply a 'free-style' vertical 'lift' force to a horizontal bar located 150 mm from the ground. Forces were applied with the horizontal location of the midpoint of the ankles at 200, 400, 600 and 800 mm from the bar. The results were compared with the Maximal Permissible Limit (MPL) and the Action Limit (AL) values proposed in the NIOSH guide. The results showed that the mean MVIS of the Hong Kong subjects lay between the MPL and AL when the load was 200 mm and 400 mm from the ankles, but was significantly below the AL at 600 mm. At 800 mm more than half of the subjects were unable to apply any measurable positive vertical force. The compressive force at the L5/S1 disc of four subjects was estimated using a static biomechanical model. The relationship between the disc pressure and the force exerted was largely in accordance with NIOSH guide values. The results are discussed in terms of differences in subject population and lifting technique. Comments on the applicability of the NIOSH standards internationally, are also presented.  相似文献   
80.
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