Fusion of lipid bilayers: a model involving mechanistic connection to HII phase forming lipids.

A model for the molecular mechanism of the fusion of lipid bilayers is described. A crucial feature of this model and related to the lamellar-->hexagonal phase HII transition is a novel, hypothetical lipid conformation, tentatively referred to here as extended. During fusion this conformation could manifest itself in the contact site between two vesicles in close proximity and involves the extension of the acyl chains of a phospholipid molecule in opposite directions, i.e. embedded into the two opposing bilayers while maintaining the headgroup in the interface. Although evidence for the occurrence of the extended conformation for phospholipids is sparse this conformation appears to be compatible with currently available experimental data. Of importance also is that the extended conformation allows for the fusion of two bilayer membranes to proceed with minimal exposure of the lipid hydrocarbon chains to water. It can also account for other features of membrane fusion such as lipid mixing in the intermediate state without mixing of the vesicle contents as well as for the molecular basis of the action of fusogenic lipids.     

Mechanism of the development of functional hyperemia in the palatal salivary glands of the rat

At various regimens of the secretory activity in the palatile salivary glands, changes occurring in the transversal profiles of the postcapillary venules, but not of the blood capillaries, are most noticeable. Under food stimulation of secretion, the former dilate essentially, that can demonstrate certain functional hyperemia developing in the palatine salivary glands. Some previous experiments concerning interpretation of principles on the microcirculatory bed spatial organization give a good reason to suggest that dilatation of the postcapillary venules is connected with an increased blood perfusion in the canals of the preferrable blood stream. The postcapillary dilatation is possible because blood from the precapillaries gets into the capacitance blood microvessels and its volume at that moment is greater than the capacity of the venous microvessels. A suggestion is made that filtrative function of the palatile salivary glands depends on development of the functional hyperemia. It is possible, that this mechanism is universal, since owing to it, reflectory reactions of the salivary glands directed to the immediate secure of the oral cavity with a necessary amount of liquor become possible.     

Characterisation of Trypanosoma cruzi populations by dna polymorphism of the cruzipain gene detected by single-stranded dna conformation polymorphism (SSCP) and direct sequencing

Fifty fresh isolates of Trypanosoma cruzi from Triatoma dimidiata vectors and 31 from patients with Chagas disease were analysed for DNA polymorphisms within the 432-bp core region of the cruzipain gene which encodes the active site of cathepsin L-like cystein proteinase. The cruzipain gene showed signs of polymorphism consisting of four different DNA sequences in Central and South American isolates of T. cruzi. The PCR fragments of Guatemalan isolates could be divided into three groups, Groups 1, 2 and 3, based on different patterns of single-stranded DNA conformation polymorphism. All of the strains isolated from Brazil, Chile, and Paraguay, except for the CL strain, showed a Group 4 pattern. Two to four isolates from each group were analysed by cloning and sequencing. A silent mutation occurred between Groups 1 and 2, and five nucleotides and two aa substitutions were detected between Groups 1 and 3. The DNA sequence of Group 4 contained five nucleotides and one aa substitution from Group 1. All of the DNA sequences corresponded well with the single-stranded DNA conformation polymorphism. The Group 1 isolates, the majority in the Guatemalan population (70/81, 86.4%), were isolated from both triatomines and humans, but Group 3 were isolated only from humans. Moreover, the Group 2 isolates were detected only in triatomine vectors (9/50; 18%), but never in humans (0/32, P<0.05) suggesting that this group has an independent life-cycle in sylvatic animals and is maintained by reservoir hosts other than humans.     

Trichoderma reesei sequences that bind to the nuclear matrix enhance transformation frequency

Three DNA fragments, trs1, 2 and 3, were isolated from the Trichoderma reesei genome on the basis of their ability to promote autonomous replication of plasmids in Saccharomyces cerevisiae. Each trs element bound specifically to the isolated T. reesei nuclear matrix in vitro, and two of them bound in vivo, indicating that they are matrix attachment regions (MARs). A similar sequence previously isolated from Aspergillus nidulans (ans1) was also shown to bind specifically to the T. reesei nuclear matrix in vitro. The T. reesei MARs are AT-rich sequences containing 70%, 86% and 73% A+T over 2.9, 0.8 and 3.7 kb, respectively for trs1, 2 and 3. They exhibited no significant sequence homology, but were shown to contain a number of sequence motifs that occur frequently in many MARs identified in other eukaryotes. However, these motifs occurred as frequently in the trs elements as in randomly generated sequences with the same A+T content. trs1 and 3 were shown to be present as single copies in the T. reesei genome. The presence of the trs elements in transforming plasmids enhanced the frequency of integrative transformation of T. reesei up to five fold over plasmids without a trs. No evidence was obtained to suggest that the trs elements promoted efficient replication of plasmids in T. reseei. A mechanism for the enhancement of transformation frequency by the trs elements is proposed. Received: 1 March 1997 / Accepted: 13 May 1997