Somatic Mutations Associated with Metastasis in Acral Melanoma

Molecular Biology - Acral melanoma is one of the most aggressive and fast-growing forms of cutaneous melanoma and is characterized by a predominant location on the palms and feet. Primary tumors,...     

State of the NPY-ergic System of the Hypothalamic Arcuate Nucleus in Rats with Experimentally Induced Diabetes Mellitus

We have carried out a quantitative study of the state of a neuropeptide Y (NPY)-ergic system of the hypothalamic arcuate nucleus (AN) in rats in the norm and in streptozotocin (STZ)-induced diabetes mellitus. The NPY-containing objects were identified using an indirect immunofluorescent technique; a system of digital analysis of images and a special software were used. Analysis of the morphometric parameters of the NPY-immunopositive objects within the AN sections allowed us to classify in an automatic mode such objects as neurons of various sizes, fibers, and terminals, as well as to count the number of objects of each class. In addition, the fluorescent intensity of the objects under study (directly proportional to the concentration and amounts of NPY in these objects) was determined. On the basis of these parameters, criteria of the state of synthesis and secretion of NPY and NPY-ergic innervation in the AN subnuclei were proposed. In control animals, the greatest number of NPY-immunopositive neurons and total content of NPY in the neurons and terminals were observed in the ventrolateral and dorsomedial subnuclei of the AN (vl- and dmAN, respectively); this fact points to the high intensities of NPY synthesis and secretion in these structures. The greatest content of NPY in the nerve terminals within the vlAN zone is indicative of the highest intensity of NPY-ergic afferent influences on this subnucleus. It is concluded that the approach we proposed is adequate enough for integral quantitative estimation of the state of peptidergic systems. The development of STZ-induced diabetes mellitus caused changes in the NPY-ergic system in different AN subnuclei; such changes demonstrated certain specificity. In particular, an increase in the number of immunopositive neurons and the total content of NPY in the neurons of vlAN and the ventromedial AN subnucleus (vmAN), which is evidence in favor of intensification of synthesis of the neuropeptide, was not accompanied by an adequate rise in the NPY content in the terminals located in the subnucleus under study. The content of NPY in the eminentia medianus also decreased. The above data allow us to hypothesize that the NPY-ergic system of the AN under conditions of diabetes mellitus is functionally insufficient.     

Expression of the envelope antigen F1 of Yersinia pestis is mediated by the product of caf1M gene having homology with the chaperone protein PapD of Escherichia coli.

The effective synthesis of the envelope antigen F1 of Y. pestis in E. coli HB101 is mediated by the expression of the caf1M gene. This gene was sequenced, and the protein encoded was found to have a significant homology with the chaperone protein PapD of uropathogenic E. coli. The data presented allow one to suppose Caf1M and PapD proteins perform similar functions in the biogenesis of the Y. pestis capsule and E. coli P-pili, respectively.     

Intracellular serine protease of Bacillus subtilis: sequence homology with extracellular subtilisins.

Intracellular serine protease was isolated from stationary-grown Bacillus subtilis A-50 cells and purified to homogeneity. The molecular weight of the enzyme is 31,000 +/- 1,000, with an isoelectric point of 4.3. Its amino acid composition is characteristically enriched in glutamic acid content, differing from that of extra-cellular subtilisins. The enzyme is completely inhibited with phenylmethylsulfonyl fluoride and ethylenediaminetetraacetic acid. Intracellular protease possesses negligible activity towards bovine serum albumin and hemoglobin, but has 5- to 20-fold higher specific activity against p-nitroanilides of benzyloxycarbonyl tripeptides than subtilisin BPN'. Esterolytic activity of the enzyme is also higher than that of subtilisin BPN'. The enzyme is sequence homologous with secretory subtilisins throughout 50 determined NH2-terminal residues, indicating the presence of duplicated structural genes for serine proteases in the B. subtilis genome. The occurrence of two homologous genes in the cell might accelerate the evolution of serine protease not only by the loosening of selective constrainst, but also by creation of sequence variants by means of intragenic recombination. Three molecular forms of intracellular protease were found, two of them with NH2-terminal glutamic acid and one minor form, three residues longer, with asparagine as NH2 terminus. These data indicate the possible presence of an enzyme precursor proteolytically modified during cell growth.     

Eu-heterochromatic rearrangements induce replication of heterochromatic sequences normally underreplicated in polytene chromosomes of Drosophila melanogaster

In polytene chromosomes of D. melanogaster the heterochromatic pericentric regions are underreplicated (underrepresented). In this report, we analyze the effects of eu-heterochromatic rearrangements involving a cluster of the X-linked heterochromatic (Xh) Stellate repeats on the representation of these sequences in salivary gland polytene chromosomes. The discontinuous heterochromatic Stellate cluster contains specific restriction fragments that were mapped along the distal region of Xh. We found that transposition of a fragment of the Stellate cluster into euchromatin resulted in its replication in polytene chromosomes. Interestingly, only the Stellate repeats that remain within the pericentric Xh and are close to a new eu-heterochromatic boundary were replicated, strongly suggesting the existence of a spreading effect exerted by the adjacent euchromatin. Internal rearrangements of the distal Xh did not affect Stellate polytenization. We also demonstrated trans effects exerted by heterochromatic blocks on the replication of the rearranged heterochromatin; replication of transposed Stellate sequences was suppressed by a deletion of Xh and restored by addition of Y heterochromatin. This phenomenon is discussed in light of a possible role of heterochromatic proteins in the process of heterochromatin underrepresentation in polytene chromosomes.     

Genetic effects of low-dose chronic irradiation of developing seeds of Arabidopsis thaliana Heynh

A six-year study, starting in 1987, focused on the frequency of embryonic lethals and chlorophyll mutations that arose in developing seeds of Arabidopsis thaliana growing at sites varying in the level of radioactive contamination in the 30-km control region of the Chernobyl Atomic Power Plant. The dose rate of chronic irradiation varied from 200 microR/h to 2.4 R/h. To study the genetic effects of various levels of radioactive contamination, the frequency of mutations arising in a particular generation was determined and the irradiation dose was estimated for the given generation. The dose dependence of the mutation frequency proved to follow a power function with a power less than unity, suggesting a relatively high effect for low-dose irradiation. Possible explanations of this phenomenon are discussed.