Pull-Down of Metalloproteins in Their Native States by Using Desthiobiotin-Based Probes

One-third of all proteins are estimated to require metals for structural stability and/or catalytic activity. Desthiobiotin probes containing metal binding groups can be used to capture metalloproteins with exposed active-site metals under mild conditions so as to prevent changes in metallation state. The proof-of-concept was demonstrated with carbonic anhydrase (CA), an open active site, Zn²⁺-containing protein. CA was targeted by using sulfonamide derivatives. Linkers of various lengths and structures were screened to determine the optimal structure for capture of the native protein. The optimized probes could selectively pull down CA from red blood cell lysate and other protein mixtures. Pull-down of differently metallated CAs was also investigated.     

Statistical analysis of subthreshold leakage current for VLSI circuits

We develop a method to estimate the variation of leakage current due to both intra-die and inter-die gate length process variability. We derive an analytical expression to estimate the probability density function (PDF) of the leakage current for stacked devices found in CMOS gates. These distributions of individual gate leakage currents are then combined to obtain the mean and variance of the leakage current for an entire circuit. We also present an approach to account for both the inter- and intra-die gate length variations to ensure that the circuit leakage PDF correctly models both types of variation. The proposed methods were implemented and tested on a number of benchmark circuits. Comparison to Monte Carlo simulation validates the accuracy of the proposed method and demonstrates the efficiency of the proposed analysis method. Comparison with traditional deterministic leakage current analysis demonstrates the need for statistical methods for leakage current analysis.     

Acculturation, Familism, Parental Monitoring, and Knowledge as Predictors of Marijuana and Inhalant Use in Adolescents.

The authors investigated relationships between marijuana and inhalant use and several cultural and demographic factors in Anglo American and Hispanic American adolescents (N=1,094). Outcome measures assessed lifetime and 30-day marijuana and inhalant use. Predictors and covariates used in logistic regression analyses were region, grade, gender, knowledge, acculturation, familism, and parental monitoring. Hispanic Americans exhibited higher usage across all measures. In this group, high acculturation was associated with low marijuana, but high inhalant, use. Across all participants, positive family relations and parental monitoring were strongly associated with attenuated marijuana use hut only among those most knowledgeable about drugs. Familism and monitoring were not associated with diminished usage among the less knowledgeable. For inhalants, monitoring combined with high knowledge or high familism was associated with diminished usage. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Effect of grain boundary phase on the thermal conductivity of aluminium nitride ceramics

AIN with high thermal conductivity was fabricated by pressureless sintering with Y₂O₃ as the sintering aid. The thermal conductivity was observed to increase with sintering time (up to 8 h) at 1810 °C. The distribution of the sintering aid was identified as one of the major factors influencing the thermal conductivity in AIN. Non-uniform distribution of the grain boundary phase was found to be associated with a significant amount of porosity, resulting in the enhancement of phonon scattering and thereby lowering the thermal conductivity.     

Nanomechanics and Raman spectroscopy of fibrillin 2 knock-out mouse bones

Absence of fibrillin 2 (Fbn2), a non-collagenous bone protein, causes a connective tissue disorder called congenital contractural arachnodactyly (CCA) and has been associated with decreased bone mineral density. Nanoindentation and Raman microspectroscopy have been used to compare the mechanical and chemical properties of cortical bone from femora of Fbn2−/− deficient mice and their wild-type controls (Fbn2+/+). It was found that Fbn2−/− bones have significantly lower hardness and elastic modulus compared to Fbn2+/+ bones, especially in the mid-cortical section. The Raman analysis showed little difference with genotype except for a decrease in type-B carbonate substitution in the endosteal region of Fbn2−/− bones. The results indicate that Fbn2 plays a direct role in determining the mechanical properties of bone.     

Neutrophil attractant protein-1 and monocyte chemoattractant protein-1 in human serum. Effects of intravenous lipopolysaccharide on free attractants, specific IgG autoantibodies and immune complexes

We recently found that normal human sera contain IgG antibodies against two chemoattractants, neutrophil attractant protein-1 (NAP-1/IL-8) and monocyte chemoattractant protein-1 (MCP-1), as well as immune complexes of these proteins. Intravenously administered LPS was reported to cause a sharp rise in serum NAP-1 concentration. Our study was designed to determine if LPS also caused an increase in MCP-1 and to measure associated changes in concentrations of antibody and immune complex. LPS caused a rise to peak within 2 to 3 h in serum concentrations of free NAP-1 and MCP-1, followed by an almost equally rapid fall toward base-line levels by about 5 h postinjection. MCP-1 concentration in sera from the 11 subjects rose to a peak of 330 +/- 52 pM. The peak value for NAP-1 was 80 +/- 11 pM. In 10 of the 11 subjects, free IgG autoantibody to MCP-1 decreased from a mean pre-LPS value of 1820 +/- 660 pM to a mean low of 53% of the respective initial values. Corresponding data for IgG anti-NAP-1 were a pre-LPS concentration of 216 +/- 7 pM, which decreased to a mean low of 44% of the respective initial values. The finding in some subjects of a rapid rise in free antibody after the nadir suggests the possibility of acute regulation of autoantibody secretion rates. Although the results suggested that LPS-induced chemoattractant combined with free antibody, serum concentrations of MCP-1-IgG or NAP-1-IgG did not increase, which points to an as yet unknown mechanism for trapping and elimination of the immune complexes.     

Sucrose and fructose have qualitatively different flavors to rats

Previous studies suggest that rats might be able to discriminate between sucrose and fructose, but no previous study has examined this possibility in much detail. Rats were conditioned to avoid either sucrose or fructose by injecting them with lithium chloride when they drank these substances. Control rats were given the same injections but were not exposed to either sugar during training. After training, the rats were given a choice of fructose vs. sucrose. Data from control rats provided information about the relative taste intensity of the sugars. If the sugars possess only a single gustatory quality, control rats should prefer the sweeter sugar; under this assumption, sucrose appears to be two-four times sweeter than fructose. The two sugars share a common taste because rats trained to avoid sucrose avoided fructose when the fructose concentration was much greater than the sucrose concentration. Nevertheless, the two sugars are discriminable because, when the apparent sweetness of the sugars was matched, rats showed a greater aversion to the sugar they were trained to avoid. Aversions to sucrose and fructose also generalized to maltodextrins, but sucrose may have a somewhat greater maltodextrin flavor than does fructose. It is proposed that the biological function of maltodextrin taste is to allow animals to sense the ratio of glucose to fructose in foods.