M-Cadherin-Mediated Cell Adhesion and Complex Formation with the Catenins in Myogenic Mouse Cells

M-cadherin is a member of the multigene family of calcium-dependent intercellular adhesion molecules, the cadherins, which are involved in morphogenetic processes. Amino acid comparisons between M-cadherin and E-, N-, and P-cadherin suggested that M-cadherin diverged phylogenetically very early from these classical cadherins. It has been shown that M-cadherin is expressed in prenatal and adult skeletal muscle. In the cerebellum, M-cadherin is present in an adherens-type junction which differs in its molecular composition from the E-cadherin-mediated adherens-type junctions. These and other findings raised the question of whether M-cadherin and the classical cadherins share basic biochemical properties, notably the calcium-dependent resistance to proteolysis, mediation of calcium-dependent intercellular adhesion, and the capability to form M-cadherin complexes with the catenins. Here we show that M-cadherin is resistant to trypsin digestion in the presence of calcium ions but at lower trypsin concentrations than E-cadherin. When ectopically expressed in LMTK⁻cells, M-cadherin mediated calcium-dependent cell aggregation. Finally, M-cadherin was capable of forming two distinct cytoplasmic complexes in myogenic cells, either with α-catenin/β-catenin or with α-catenin/plakoglobin, as E- and N-cadherin, for example, have previously been shown to form. The relative amount of these complexes changed during differentiation from C2C12 myoblasts to myotubes, although the molecular composition of each complex was unaffected during differentiation. These results demonstrate that M-cadherin shares important features with the classical cadherins despite its phylogenetic divergence.     

The taste of polycose in hamsters

Hamsters show a preference for Polycose, a mixture of starch-derived glucose polymers, that is as strong as their preference for sucrose. However, in the hamster, taste aversions to Polycose may be less easily acquired than taste aversions to sucrose and the qualitative aspects of Polycose are unknown in this species. In order to examine the taste of Polycose in the hamster, we utilized a taste-aversion protocol with two conditioning trials. Animals were trained to avoid one of three different conditioning stimuli: 50 mM sucrose, 100 mM Polycose and a mixture of 50 mM sucrose with 100 mM Polycose. Control animals were conditioned with deionized water. After the second conditioning trial, generalization testing began for the three conditioning stimuli plus 3 mM citric acid, 300 mM KCI and 30 mM NaCl. The results showed that aversions to Polycose, sucrose or the Polycose/sucrose mixture cross- generalized, demonstrating that Polycose and sucrose share a common taste percept in the hamster. None of the aversions generalized to NaCl, citric acid or KCI. In addition, comparisons among the patterns of taste generalizations indicated that the tastes of Polycose and sucrose also had distinct qualitative components. Finally, although the taste of 100 mM Polycose was more salient than the taste of 50 mM sucrose, the taste of sucrose could still be detected in a mixture with Polycose.      

Production of Ochratoxins in Different Cereal Products by Aspergillus ochraceus

The effects of temperature and length of incubation on ochratoxin A production in various substrates were studied. The optimal temperature for toxin production by Aspergillus ochraceus NRRL-3174 was found to be around 28 C. Very low levels of ochratoxin A are produced in corn, rice, and wheat bran at 4 C. The optimal time for ochratoxin A production depends on the substrate, ranging from 7 to 14 days at 28 C. Ochratoxin B and dihydroisocoumaric acid, i.e., one of the hydrolysis products of ochratoxin A, were produced in rice but at levels considerably lower than ochratoxin A. No ochratoxin C was produced in rice at 28 C. When added to rice cereal or oatmeal, the toxin was found to be very stable over prolonged storage and even to autoclaving for 3 hr.     

Nitrate influx and efflux by intact wheat seedlings: Effects of prior nitrate nutrition

Summary Wheat (Triticum vulgare L., cv. Blueboy) seedlings, grown with 0.25, 1.0 and 15 mM nitrate in complete nutrient solutions, were transferred 10 days after germination to 1.0 mM K¹⁵NO₃ (99 A% ¹⁵N) plus 0.1 mM CaSO₄ at pH 6.0. The solutions were replaced periodically over a 6-h period (5 mW cm^-2; 23°). Changes in the [¹⁵N]- and [¹⁴N]nitrate in the solution were determined by nitrate reductase and mass-spectrometric procedures and potassium by flame photometry. Influx of [¹⁵N]nitrate was depressed in plants grown at 1.0 mM nitrate relative to those grown at 0.25 mM, but there was no appreciably difference in [¹⁴N]nitrate efflux. Prior growth at 15 mM further restricted [¹⁵N]nitrate influx which, together with a substantial increase in [¹⁴N]nitrate efflux, resulted in no net nitrate uptake during the course of the experiment. Efflux of [¹⁴N]nitrate occurred to solutions containing no nitrate but it was significantly enhanced upon exposure to [¹⁵N]nitrate in the external solution. Influx of [¹⁵N]nitrate was more restricted at 5°, relative to 23°, than was [¹⁴N]nitrate efflux. The nitrate concentrations of the root tissue immediately before exposure to the K¹⁵NO₃ solutions did not give a precise indication of the subsequent [¹⁵N]nitrate influx rates nor of the [¹⁴N]nitrate efflux rates. Net K⁺ uptake was related to the magnitude of the net nitrate uptake, not to the initial K⁺ concentration in the roots. The data are interpreted as indicating that [¹⁵N]nitrate influx and [¹⁴N]nitrate efflux are largely independent processes, subject to different controls, and that net nitrate uptake provides the driving force for net potassium uptake.Paper No. 4884 of the Journal Series of the North Carolina Agricultural Experiment Station, Raleigh, NC, USA. This investigation was supported in part by the U.S. Energy Research and Development Administration, Contract No. AT-(40-1)-2410     

Purification of galectin-3 from ovine placenta: developmentally regulated expression and immunological relevance

Galectins, beta-galactoside-binding lectins, are extensively distributed in the animal kingdom and share some basic molecular properties. Galectin-3, a member of this family, is generally associated with differentiation, morphogenesis, and metastasis. In this study, galectin-3 was isolated from ovine placental cotyledons round the middle of the gestation period by lactose extraction followed by affinity chromatography on lactosyl-agarose, and separated from galectin-1 by size exclusion chromatography on a Superose 12 column. Under native conditions this lectin behaved as a monomer with an apparent molecular weight of approximately 29,000 and an isoelectric point of 9.0. The partial amino acid sequence of the peptides obtained by tryptic digestion of this protein followed by HPLC separation showed striking homology with other members of the galectin-3 subfamily. Furthermore, ovine placental galectin-3 exhibited specific mitogenic activity toward rat spleen mononuclear cells. Besides, this protein strongly reacted with a rabbit antiserum raised against a chicken galectin. Results obtained by Western blot analysis showed that its expression was greatly decreased in term placenta with respect to the middle of the gestation period, suggesting a regulated expression throughout development.      

The subcellular localizations of atypical synaptotagmins III and VI. Synaptotagmin III is enriched in synapses and synaptic plasma membranes but not in synaptic vesicles.

Multiple synaptotagmins are expressed in brain, but only synaptotagmins I and II have known functions in fast, synchronous Ca2+-triggered neurotransmitter release. Synaptotagmin III was proposed to regulate other aspects of synaptic vesicle exocytosis, particularly its slow component. Such a function predicts that synaptotagmin III should be an obligatory synaptic vesicle protein, as would also be anticipated from its high homology to synaptotagmins I and II. To test this hypothesis, we studied the distribution, developmental expression, and localization of synaptotagmin III and its closest homolog, synaptotagmin VI. We find that synaptotagmins III and VI are present in all brain regions in heterogeneous distributions and that their levels increase during development in parallel with synaptogenesis. Furthermore, we show by immunocytochemistry that synaptotagmin III is concentrated in synapses, as expected. Surprisingly, however, we observed that synaptotagmin III is highly enriched in synaptic plasma membranes but not in synaptic vesicles. Synaptotagmin VI was also found to be relatively excluded from synaptic vesicles. Our data suggest that synaptotagmins III and VI perform roles in neurons that are not linked to synaptic vesicle exocytosis but to other Ca2+-related nerve terminal events, indicating that the functions of synaptotagmins are more diverse than originally thought.     

A biomechanical analysis of finger joint forces and stresses developed during common daily activities

The problem of modelling stresses incurred at the finger joints is critical to the design of durable joint replacements in the hand. The goal of this study was to characterise the forces and stresses at the finger and thumb joints occurring during activities such as typing at a keyboard, playing piano, gripping a pen, carrying a weight and opening a jar. The metacarpal and proximal phalanx were modelled using a COMSOL-based finite element analysis. Analysis of these activities indicates that joint forces in excess of 100 N may be common at the metacarpophalangeal joint (MCP) due to carrying objects such as groceries or while opening jars. The model predicted that stresses in excess of 2 MPa, similar to stresses at the hip, occur at the MCP with the properties of cancellous bone playing a significant role in the magnitude and distribution of stress.     

黄檗丛枝菌根真菌鉴定

目的：利用形态学特征与Nested-PCR技术鉴定黄檗丛枝菌根真菌。方法：采用酸性品红染色法挑选黄檗丛枝菌根。同时,利用湿筛法获得AM真菌孢子,进行形态学鉴定。运用Nested-PCR技术,对黄檗粗提DNA进行特异性扩增,采用blastn进行序列相似性比较。并构建系统进化树,确定侵染黄檗根系的AM真菌。结果：编号为HDAM-1的AM真菌孢子,形态特征与G．intraradices的特征描述一致。Nested-PCR检测到约455bp的目的片段,其序列与G．intraradices（DQ469118）相似性最高,达97．8％,有11个碱基的差异。系统进化树显示该序列在基于25S rDNA的进化树中与G．intraradices（DQ469118．1）处于同一分支,确定G．intraradices侵染黄檗根系。结论：将形态学特征与Nested-PCR技术相结合鉴定AM真菌,不仅简易、经济,而且能够提高研究结果的可靠性。