Binding kinetics of monoclonal antibody using antigen-beta-galactosidase hybrid protein: application to measurement of peptide antigenicity

A simple method for determination of binding kinetics of a solid-phase antibody using antigen-beta-galactosidase hybrid protein was evaluated. To minimize conformational change of the antigen binding site of the antibody when directly binding to a microtiter plate, the microtiter plate was precoated with protein A. The binding and free antigen concentrations were directly obtained from the beta-galactosidase activity. This method can be used for analyses of the equilibrium dissociation constant (KD), and the association (Kass) and dissociation (Kdiss) rate constants. Peptide antigenicity was also analyzed by competitive ELISA using this method. Since both antigen-beta-galactosidase and the peptide used are localized in the fluid-phase, the proper affinity constant (KA) of the peptide can be estimated from the KD value of the antigen-beta-galactosidase-antibody interaction, and from the IC50 value of the peptide.     

Aminopeptidase N purified from gypsy moth brush border membrane vesicles is a specific receptor for Bacillus thuringiensis CryIAc toxin

We have evaluated the binding of Bacillus thuringiensis Cry toxins to aminopeptidase N (APN) purified from Lymantria dispar (gypsy moth) brush border membrane vesicle (BBMV). CryIAc toxin bound strongly to APN, while either the structurally related CryIAa and CryIAb toxins or CryIC, CryIIA, and CryIIIA toxins showed weak binding to APN. An in vitro competition binding study demonstrated that the binding of CryIAc to L. dispar BBMV was inhibited by APN. Inhibition of short circuit current for CryIAc, measured by voltage clamping of whole L. dispar midgut, was substantially reduced by addition of phosphatidylinositol-specific phospholipase C, which is known to release APN from the midgut membrane. In contrast, addition of phosphatidylinositol-specific phospholipase C had only a marginal effect on the inhibition of short circuit current for CryIAa. These data suggest that APN is the major functional receptor for CryIAc in L. dispar BBMV. A ligand blotting experiment demonstrated that CryIAc recognized a 120-kDa peptide (APN), while CryIAa and CryIAb recognized a 210-kDa molecule in L. dispar BBMV. In contrast, CryIAa and CryIAb bound to both the 120- and 210-kDa molecules in Manduca sexta BBMV, while CryIAc recognized only the 120-kDa peptide. The 120-kDa peptide (APN) in L. dispar BBMV reacted with soybean agglutinin, indicating that N-acetylgalactosamine is a component of this glycoprotein.     

Prognosis versus actual outcome. III. The effectiveness of clinical parameters in accurately predicting tooth survival

Tooth loss for 100 treated periodontal patients (2,509 teeth) under maintenance care was evaluated to determine the effectiveness of commonly taught clinical parameters utilized in the assignment of prognosis in accurately predicting tooth survival. Previous studies in this series evaluated prognosis as a surrogate variable representing the condition of the tooth at a particular point. In this study, survival analysis was used to evaluate the relationship of these common clinical parameters to an actual end point, tooth loss. Robust log rank tests indicated that initial probing depth, initial furcation involvement, initial mobility, initial crown-to-root ratio, and initial root form were all associated with tooth loss. In addition, smoking and increased initial bone loss were both found to be associated with increased risk of tooth loss while fixed abutment status was associated with a decreased risk of tooth loss. A Cox proportional hazards regression model showed that initial probing depth, initial furcation involvement, initial mobility, initial percent bone loss, presence of a parafunctional habit without a biteguard, and smoking were all associated with an increased risk of tooth loss. This model suggests that patients are twice as likely to loose their teeth if there is increasing mobility, if they have a parafunctional habit and do not wear a biteguard, or if they smoke. From these data there does appear to be a relationship between the assigned prognosis and tooth loss. Teeth with worse prognosis have a worse survival rate, but the commonly taught clinical parameters used in the traditional method of assignment of prognosis do not adequately explain that relationship. Furthermore, initial prognosis did not adequately explain the condition of the tooth or accurately predict the tooth's survival. These results seem to indicate that the effect of these clinical parameters on tooth survival is only partially reflected in the assigned prognosis initially, suggesting that perhaps some of the clinical parameters should be weighed more heavily than others when assigning prognosis. Further studies are needed to develop a more accurate method for the assignment of prognosis.     

A scaled, high-performance (4.5 fJ) bipolar device in a 0.35 μmhigh-density BiCMOS SRAM technology

We present the performance improvements obtained both by scaling the Selectively Compensated Collector (SCC) BJT and by using a modified Current-Mode Logic (CML) gate configuration. Scaling the perimeter parameter by using the (tighter) bitcell design rules results in a ~30% reduction in parasitic capacitances, and a 23% lower power-delay product; reducing it from 48 fJ to 37 fJ. The greatest return comes from using a modified CML gate, which has an n-MOS current source. At a supply voltage of 1.1 V, and at 40 μA switching current, the minimum power-delay product of this CML gate is a silicon-substrate bipolar record 4.5 fJ     

Three sibs with mild variety of osteopetrosis

We report three sibs with mild autosomal recessive variety of osteopetrosis. The prominent clinical features were short stature, malocclusion of teeth, hepatosplenomegaly and a typical facial appearance. The only atypical features were microcephaly, a normal upper segment to lower segment ratio and a normal arm span.     

Glutathione-binding proteins of Setaria digitata: antibody responses in human infected with Wuchereria bancrofti

Glutathione-s-transferase activity was determined in filarial parasites. The activity was detected in adult stages of cattle parasite Setaria digitata. It was absent in other stages of Setaria and also in infective larval stages of Wuchereria bancrofti and Brugia malayi. The activity was enhanced about twenty five fold following purification of adult setaria extracts on glutathione agarose column. Antibody (IgG and IgM) levels to the affinity purified proteins (SdGBP) were detected predominantly (90%) in Wuchereria bancrofti infected individuals compared with normal residents of endemic regions. IgA and IgE responses could not be detected. Filarial sera in contrast to non-filarial caused reduction in the enzymatic activities of Sd GBP. Microfilaraemic sera after diethylcarbamazine treatment resulted in enhanced reduction of enzymatic activity.     

Linkage map of Escherichia coli K-12, edition 10: the traditional map

This map is an update of the edition 9 map by Berlyn et al. (M. K. B. Berlyn, K. B. Low, and K. E. Rudd, p. 1715-1902, in F. C. Neidhardt et al., ed., Escherichia coli and Salmonella: cellular and molecular biology, 2nd ed., vol. 2, 1996). It uses coordinates established by the completed sequence, expressed as 100 minutes for the entire circular map, and adds new genes discovered and established since 1996 and eliminates those shown to correspond to other known genes. The latter are included as synonyms. An alphabetical list of genes showing map location, synonyms, the protein or RNA product of the gene, phenotypes of mutants, and reference citations is provided. In addition to genes known to correspond to gene sequences, other genes, often older, that are described by phenotype and older mapping techniques and that have not been correlated with sequences are included.