ATP-Mediated Compositional Change in Peripheral Myelin Membranes: A Comparative Raman Spectroscopy and Time-Of-Flight Secondary Ion Mass Spectrometry Study

In the present paper we addressed a mechanism of the myelin reorganization initiated by extracellular ATP and adenosine in sciatic nerves of the frog Rana temporaria. In combination with Raman microspectroscopy, allowing noninvasive live-cell measurements, we employed time-of-flight secondary ion mass spectrometry (TOF-SIMS) to follow the underlying changes in chemical composition of myelin membranes triggered by the purinergic agents. The simultaneous increase in lipid ordering degree, decrease in membrane fluidity and the degree of fatty acid unsaturation were induced by both ATP and adenosine. Mass spectrometry measurements revealed that ATP administration also led to the marked elevation of membrane cholesterol and decrease of phosphotidylcholine amounts. Vesicular lipid transport pathways are considered as possible mechanisms of compositional and structural changes of myelin.     

Lipid and pigment changes during shoot initiation in cultured explants of Pinus radiata

Cotyledon explants of radiata pine contain principally lipid and protein reserve materials, which decline during shoot initiation. This process was followed phytochemically and ultrastructurally. Fatty acid and sterol analyses indicated that there were both quantitative and qualitative changes in the different classes of lipid. The most pronounced changes were an increase in the linolenic acid content of the polar lipids and the appearance of stigmasterol during shoot initiation. There was also a continued increase in chlorophyll and carotenoid levels, which paralleled chloroplast development. It appears that the changes observed were similar to those that occur in cotyledons during normal seedling development.     

Massenanzucht phototropher Organismen in einer automatischen Kulturanlage

Summary A new apparatus is described which permits pure cultivation of 110 l amounts of phototrophic organisms. Some data are given to prove its utility for mass production (kilogram amounts) of algae and photosynthetic bacteria.     

Specific refractoriness of adenylate cyclase in skin to epinephrine,prostaglandin E,histamine and AMP

The cyclic AMP level in pig skin (epidermis) increases markedly after incubation with epinephrine, prostaglandin E, histamine or adenosine 5′-monophosphate. This increase is transient and “spiking” is the consistent response to these four stimulators. The “spiking” is due to a non-responsiveness or refractoriness which develops within minutes and is specific to any one stimulating hormone but not to the others. The addition of inhibitors of protein syntheses did not prevent the development of the refractoriness. Adenylate cyclase and phosphodiesterase activities measured in skin homogenates prepared from skin samples taken before, during and after the “spiking” did not change significantly. The hormone-induced refractoriness in this skin system appears to be due to a specific, localized loss of function of the adenylate cyclase system.     

A Single Residue in a Novel ADP-ribosyl Cyclase Controls Production of the Calcium-mobilizing Messengers Cyclic ADP-ribose and Nicotinic Acid Adenine Dinucleotide Phosphate

Cyclic ADP-ribose and nicotinic acid adenine dinucleotide phosphate are ubiquitous calcium-mobilizing messengers produced by the same family of multifunctional enzymes, the ADP-ribosyl cyclases. Not all ADP-ribosyl cyclases have been identified, and how production of different messengers is achieved is incompletely understood. Here, we report the cloning and characterization of a novel ADP-ribosyl cyclase (SpARC4) from the sea urchin, a key model organism for the study of calcium-signaling pathways. Like several other members of the ADP-ribosyl cyclase superfamily, SpARC4 is a glycoprotein targeted to the plasma membrane via a glycosylphosphatidylinositol anchor. However, unlike most other members, SpARC4 shows a remarkable preference for producing cyclic ADP-ribose over nicotinic acid adenine dinucleotide phosphate. Mutation of a single residue (tyrosine 142) within a noncanonical active site reversed this striking preference. Our data highlight further diversification of this unusual enzyme family, provide mechanistic insight into multifunctionality, and suggest that different ADP-ribosyl cyclases are fine-tuned to produce specific calcium-mobilizing messengers.     

Asymmetric and Globular Forms of AChE in Slow and Fast Muscles of 129/ReJ Normal and Dystrophic Mice

Abstract: Acetylcholinesterase activities and molecular forms were studied in normal and dystrophic 129/ReJ mice, focusing on four predominantly fast-twitch muscles and the slow-twitch soleus. The asymmetric and globular forms were analyzed separately so that the effect of dystrophy on each form could be determined. This comparative study showed the following. (1) In the normal condition, each muscle exhibited a distinct distribution of the molecular forms. (2) The diversity among the fast muscles resulted mainly from variations in the proportions of the three globular forms; in contrast, these muscles showed a constant and precise A₁₂/A₈/A₄ ratio. (3) The slow-twitch soleus clearly differed from the other muscles in its low acetylcholinesterase activity and distinct distribution of the molecular forms, characterized by a low level of G₄ and a peculiar ratio among its asymmetric forms, resulting from a relative increase of the A₈ and A₄ forms. (4) In dystrophic mice, the diversity of the acetylcholin esterase distribution was lost; all the fast muscles displayed profiles exhibiting the characteristics typical of the soleus. The fast-twitch extensor digitorum longus, sternomastoid, and plantaris converged towards an identical set of acetylcholinesterase molecules. (5) In contrast, the acetylcholinesterase activity and molecular forms of the soleus were only slightly affected by the disease. These results reveal that the dystrophy modifies both categories of molecular forms of acetylcholinesterase in a very precise manner. Such complex changes, which are highly reproducible in a variety of different muscles, are unlikely to result from nonspecific reactions secondary to the disease.     

Reducing the Time Requirement of k-Means Algorithm

Traditional k-means and most k-means variants are still computationally expensive for large datasets, such as microarray data, which have large datasets with large dimension size d. In k-means clustering, we are given a set of n data points in d-dimensional space R^d and an integer k. The problem is to determine a set of k points in R^d, called centers, so as to minimize the mean squared distance from each data point to its nearest center. In this work, we develop a novel k-means algorithm, which is simple but more efficient than the traditional k-means and the recent enhanced k-means. Our new algorithm is based on the recently established relationship between principal component analysis and the k-means clustering. We provided the correctness proof for this algorithm. Results obtained from testing the algorithm on three biological data and six non-biological data (three of these data are real, while the other three are simulated) also indicate that our algorithm is empirically faster than other known k-means algorithms. We assessed the quality of our algorithm clusters against the clusters of a known structure using the Hubert-Arabie Adjusted Rand index (ARI_HA). We found that when k is close to d, the quality is good (ARI_HA>0.8) and when k is not close to d, the quality of our new k-means algorithm is excellent (ARI_HA>0.9). In this paper, emphases are on the reduction of the time requirement of the k-means algorithm and its application to microarray data due to the desire to create a tool for clustering and malaria research. However, the new clustering algorithm can be used for other clustering needs as long as an appropriate measure of distance between the centroids and the members is used. This has been demonstrated in this work on six non-biological data.