Highly potent inhibitors of TNF-alpha production. Part I: discovery of new chemical leads and their structure-activity relationships

Discovery of new chemical leads of inhibitors for TNF-alpha production starting from the chemical modification of 1 is reported. Further biological studies of 1 to disclose the site of its action strongly suggested that 1 inhibits LPS-induced TNF-alpha expression in the liver and spleen of mice. Structure-activity relationships (SARs) are also discussed and full details including the chemistry are reported.     

Functional importance of Ca2+-deficient N-terminal lobe of molluscan troponin C in troponin regulation

Ca(2+)-binding sites I and II in the N-terminal lobe of molluscan troponin C (TnC) have lost the ability to bind Ca(2+) due to substitutions of the amino acid residues responsible for Ca(2+) liganding. To evaluate the functional importance of the Ca(2+)-deficient N-terminal lobe in the Ca(2+)-regulatory function of molluscan troponin, we constructed chimeric TnCs comprising the N-terminal lobes from rabbit fast muscle and squid mantle muscle TnCs and the C-terminal lobe from akazara scallop TnC, TnC(RA), and TnC(SA), respectively. We characterized their biochemical properties as compared with those of akazara scallop wild-type TnC (TnC(AA)). According to equilibrium dialysis using (45)Ca(2+), TnC(RA), and TnC(SA) bound stoichiometrically 3 mol Ca(2+)/mol and 1 mol Ca(2+)/mol, respectively, as expected from their primary structures. All the chimeric TnCs exhibited difference-UV-absorption spectra at around 280-290 nm upon Ca(2+) binding and formed stable complexes with akazara scallop troponin I, even in the presence of 6M urea, if Ca(2+) was present. However, when the troponin complexes were constructed from chimeric TnCs and akazara scallop troponin T and troponin I, they showed different Ca(2+)-regulation abilities from each other depending on the TnC species. Thus, the troponin containing TnC(SA) conferred as high a Ca(2+) sensitivity to Mg-ATPase activity of rabbit actomyosin-akazara scallop tropomyosin as did the troponin containing TnC(AA), whereas the troponin containing TnC(RA) conferred virtually no Ca(2+) sensitivity. Our findings indicate that the N-terminal lobe of molluscan TnC plays important roles in molluscan troponin regulation, despite its inability to bind Ca(2+).     

Filopodia formation mediated by receptor tyrosine kinase Ror2 is required for Wnt5a-induced cell migration

The receptor tyrosine kinase Ror2 plays important roles in developmental morphogenesis. It has recently been shown that Ror2 mediates Wnt5a-induced noncanonical Wnt signaling by activating the Wnt-JNK pathway and inhibiting the beta-catenin-TCF pathway. However, the function of Ror2 in noncanonical Wnt signaling leading to cell migration is largely unknown. We show, using genetically different or manipulated cultured cells, that Ror2 is critical for Wnt5a-induced, but not Wnt3a-induced, cell migration. Ror2-mediated cell migration requires the extracellular cysteine-rich domain (CRD), which is the binding site for Wnt5a, and the cytoplasmic proline-rich domain (PRD) of Ror2. Furthermore, Ror2 can mediate filopodia formation via actin reorganization, irrespective of Wnt5a, and this Ror2-mediated filopodia formation requires the actin-binding protein filamin A, which associates with the PRD of Ror2. Intriguingly, disruption of filopodia formation by suppressing the expression of either Ror2 or filamin A inhibits Wnt5a-induced cell migration, indicating that Ror2-mediated filopodia formation is essential for Wnt5a-induced cell migration.     

Evidence for the presence of carbonic anhydrase III in the myoid cells of the bovine thymus. Immunocytochemical and ultrastructural study

An immunohistochemical study was carried out to detect the localization of carbonic anhydrase III (CA-III) in the bovine thymus. It was found that the CA-III activity was localized in the cells forming small clusters dispersed in the medullary region. By ultrastructural observation, these cells were identified as myoid cells.     

Designing and Judd–Ofelt evaluation of versatile luminescent Eu (III) complexes sensitized with β-diketone ligand for multifarious applications

The present research work entails the synthesis of one binary and four ternary red light−emitting Eu (III)-based complexes with 3-benzylidene-2,4-pentanedione as the main ligand and 1,10-phenanthroline, bathophenanthroline, neocuproine, and 4,4′-′dimethyl-2,2′-′bipyridyl as auxiliary ligands. The metal–organic framework of the series was elucidated using energy dispersive X-ray analysis, elemental analysis, Fourier transform infrared spectroscopy, and proton nuclear magnetic resonance. This Eu (III) series exhibits optimum thermal stability, making them a promising candidate for organic light-emitting diodes. On the basis of emission spectra, their optical parameters such as nonradiative and radiative decay rates, luminescence decay time, intrinsic quantum efficiency, and Judd–Ofelt intensity parameter were determined. The monocentric luminescence and Judd–Ofelt parameters reveal the absence of symmetry around the europium center. CIE chromaticity coordinates, correlated color temperature values, color purity, and asymmetric ratios authenticate the color coordinates of the complexes in red region. Optical band gap values lie within the range of wide band gap semiconductors, indicating their utilization in military radars and biological labeling.     

Comparative studies on heat stability and autolysis of scallop (Patinopecten yessoensis) calpain II-like proteinase and rabbit calpain II.

1. The scallop calpain-like proteinase is about five times more labile than the rabbit calpain II upon heat treatment at 35 degrees C. 2. By autolysis of the scallop proteinase of two 100 kDa subunits, 90, 45 and 30 kDa fragments were formed. Thereby the activity decreased monophasically in the presence of millimolar order of Ca2+, but did not increase in the presence of micromolar order of Ca2+ unlike the rabbit calpain II.     

Determination and developmental changes in carbonic anhydrase III in swine liver.

1. CA-III was measured by enzyme-immunoassay in the livers of male and female swine aged from the fetus to 5 years old. 2. No sexual dimorphism in porcine liver could be detected at 6 months, but stag showed twice as much as swine of the same age. 3. The concentration of CA-III in the liver increased during development up to 6 months of age, followed by decline due to senescence.     

Comparative immunohistolocalization of carbonic anhydrase isozymes I, II and III in the equine and bovine digestive tract

Summary Immunohistochemical localizations of carbonic anhydrase isozymes (CA-I, CA-II and CA-III) in equine and bovine digestive tracts were studied. In the horse, epithelial cells in both the oesophagus and non-glandular part of the stomach lacked all three isozymes. In contrast, surface epithelial and parietal cells in the glandular region of the stomach showed reactivity for CA-II. In the small intestine, absorptive columnar cells covering the villi in the duodenum were positive for CA-II. The epithelium of the jejunum and ileum lacked all three isozymes. In the large intestine, CA-II was detected in the columnar cells in the upper part of the crypt. In cattle, epithelial cells of the oesophagus showed reactions for CA-I and CA-III but not for CA-II. Although the absorptive epithelial cells of the small intestine lacked CA-I, CA-II and CA-III, those of the upper part of large intestine crypts were heavily stained for all three isozymes.