Design of novel, potent, noncovalent inhibitors of thrombin with nonbasic P-1 substructures: rapid structure-activity studies by solid-phase synthesis

Study of surface representations of the inhibitor-bound thrombin P-1 pocket revealed a lipophilic recess in this pocket which is not occupied by any known inhibitor. Solid-phase synthesis was used to generate benzylamides of D-diphenylAlaPro by aminolysis of Boc dipeptide Kaiser resin. The resulting amides inhibited thrombin in the range IC50 = 3-13,000 nM, and the structure-activity relationships and molecular modeling suggest a unique fit of the benzyl side chain into P-1 with the meta substituent occupying the recess.     

Validity of dietary recall over 20 years among California Seventh-day Adventists

Past dietary habits are etiologically important to incident disease. Yet the validity of such measurements from the previous 10-20 years is poorly understood. In this study, the authors correlated food frequency results that were obtained in 1994-1995 but pertained to recalled diet in 1974 with the weighted mean of five random 24-hour dietary recalls obtained by telephone in 1974. The subjects studied were 72 Seventh-day Adventists who lived within 30 miles of Loma Linda, California; had participated in a 1974 validation study; were still alive; and were willing to participate again in 1994. A method was developed to allow correction for random error in the reference data when these data had differentially weighted components. The results showed partially corrected correlation coefficients of greater than 0.30 for coffee, whole milk, eggs, chips, beef, fish, chicken, fruit, and legumes. Higher correlations on average were obtained when the food frequencies were scored simply 1-9, reflecting the nine frequency categories. The 95% confidence intervals for 15 of the 28 correlations excluded zero. Incorporation of portion size information was unhelpful. The authors concluded that in this population, data recalled from 20 years ago should be treated with caution but, for a number of important foods, that the degree of validity achieved approached that obtained when assessing current dietary habits.     

Small intestinal submucosa: a substrate for in vitro cell growth

The extracellular matrix (ECM) of the small intestinal submucosa (SIS) was harvested by removing the superficial layers of the mucosa and the external muscular layers. The remaining 80 microns thick sheet was disinfected and sterilized by methods which removed all cellular components. The SIS-ECM, retaining its native 3-dimensional microarchitecture and composition, was evaluated for its ability to support in vitro cell growth. Six separate cell types were seeded either alone or in coculture with other cells upon this matrix, grown in selected media, a examined daily for time periods ranging from 48 h to 2 weeks. The six cell types tested were NIH Swiss mouse 3T3 fibroblast, NIH 3T3/j2 fibroblasts, primary human fibroblasts, primary human keratinocytes, human microvascular endothelial cells (HMECs), and an established rat osteosarcoma (ROS) cell line. All cell types showed the ability to attach a proliferate. All fibroblast cell line and the keratinocytes proliferated and/or migrated into the 3-dimensional scaffold of the SIS matrix. The ROS cells and the HMECs were confined in their growth pattern to the surface of the matrix. Coculturing of NIH 3T3/J2 fibroblasts and primary human keratinocytes resulted in a distinctive spatial orientation of the two cell types. The fibroblast populated the mid-substance of the 3-dimensional matrix and the keratinocytes formed an epidermal structure with rete ridge-like formation and stratification when the composite was lifted to an air liquid interface in culture. In summary, SIS provides a substratum with a 3-dimensional scaffold that allows for cell migration and spatial organization. The substratum is suitable for in vitro studies of the interaction between epithelial or mesenchymal cells and a naturally occurring extracellular matrix.     

Placental implantation at 18 weeks and migration throughout pregnancy

BACKGROUND: We wanted to find the most frequent site of placental implantation at 18 weeks' gestation and placental migration during gestation. METHODS: Placental location was identified in 2,526 singleton pregnancies at 18 weeks' gestation and characterized into nine groups. Placental migration in 1,336 of these pregnancies was assessed by serial ultrasonography. RESULTS: At 18 weeks' gestation, posterior, high placental implantation was more common (45.1%) than anterior, high implantation (42.1%). Relocation of posterior, high placentas farther fundally (16.9%) was three times more likely than farther fundal migration of anterior, high placentas (4%). CONCLUSIONS: Posterior fundal placental implantation is more common at 18 weeks' gestation than anterior implantation. Posteriorly implanted placentas are more likely than anterior placentas to migrate farther fundally during gestation. The greater relocation of posterior placentas farther fundally suggests a greater growth of posterior versus anterior uterine wall smooth muscle.     

Count nouns, adjectives, and perceptual properties in children's novel word interpretations.

Three-year-old children were shown a novel exemplar toy and asked to judge test items that differed from the exemplar in shape, coloration, or material substance. In the count noun condition, children judged whether test items had the same novel name as the exemplar. In the adjective condition, children judged whether a test item could be described by the same novel adjective as the exemplar. The results of 3 experiments indicated that children systematically attend to shape in interpreting novel count nouns, but their interpretation of adjectives is contextually determined. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Designing Polymer Blends Using Neural Networks, Genetic Algorithms, and Markov Chains

In this paper we present a new technique to simulate polymer blends that overcomes the shortcomings in polymer system modeling. This method has an inherent advantage in that the vast existing information on polymer systems forms a critical part in the design process. The stages in the design begin with selecting potential candidates for blending using Neural Networks. Generally the parent polymers of the blend need to have certain properties and if the blend is miscible then it will reflect the properties of the parents. Once this step is finished the entire problem is encoded into a genetic algorithm using various models as fitness functions. We select the lattice fluid model of Sanchez and Lacombe (J. Polym. Sci. Polym. Lett. Ed., vol. 15, p. 71, 1977), which allows for a compressible lattice. After reaching a steady-state with the genetic algorithm we transform the now stochastic problem that satisfies detailed balance and the condition of ergodicity to a Markov Chain of states. This is done by first creating a transition matrix, and then using it on the incidence vector obtained from the final populations of the genetic algorithm. The resulting vector is converted back into a population of individuals that can be searched to find the individuals with the best fitness values. A high degree of convergence not seen using the genetic algorithm alone is obtained. We check this method with known systems that are miscible and then use it to predict miscibility on several unknown systems.     

Effect of soybean hulls, soy lecithin, and soapstock mixtures on ruminal fermentation and milk composition in dairy cows

To clarify the regulatory mechanism of pro-gelatinase A (proGelA) activation at a cellular level, expression of gelatinase A (GelA), three MT-MMPs, and TIMP-2 was examined with 11 human cancer cell lines cultured in the presence and absence of stimulants. MT1-MMP mRNA was expressed in 8 cell lines, while MT2-MMP and MT3-MMP mRNAs were expressed in fewer cell lines. The cells with high proGelA activation strongly expressed MT1-MMP mRNA but not MT2-MMP and MT3-MMP mRNAs, suggesting that MT1-MMP was responsible for the proGelA activation in the cancer cells. Treatments with concanavalin A (Con A) and a phorbor ester (TPA) enhanced the MT1-MMP expression, but only Con A stimulated the proGelA activation in many cell lines. In HT1080 fibrosarcoma cells, however, TPA also stimulated the activation. The level of TIMP-2 secreted into culture medium inversely correlated with proGelA activation. For example, 2 squamous cell carcinoma lines (HSC-3 and HSC-4) and 3 HT1080 clones, which efficiently activated proGelA, secreted little TIMP-2 into medium, whereas other cell lines and other HT1080 clones, which hardly activated proGelA, secreted TIMP-2 at high levels. When HSC-3 cells were incubated with TIMP-2 protein or transfected with TIMP-2 cDNA, the proGelA activation was strongly inhibited. These results indicated that extracellular TIMP-2 was an important negative regulator of proGelA activation. However, the level of extracellular TIMP-2 was not consistent with that of TIMP-2 mRNA in some cell lines. Other experimental results suggested that TIMP-2 might be rapidly metabolized after binding to MT1-MMP, and Con A treatment might stabilize the complex of TIMP-2 and MT1-MMP on cell membranes.