A unique DNAse associated with mitogen-induced transformation of rat lymphocytes

A unique DNAse was found, following electrophoretic separation, in rat lymphocytes stimulated with either ConA or pokeweed mitogens. This DNAse was absent or minimal in non-stimulated cells. The enzyme was active on native DNA at neutral pH and was activated by EDTA. A possible biological role is discussed as related to DNA replication.     

Cytochemical approach to the development and behaviour of rat alveolar type II cells in culture

The development and characteristics of rat alveolar type II cells were monitored by using various cytochemical techniques. Polarized light microscopy was found useful for observing live type II cells in culture. Cells progressively lose their birefringent granules starting from 48 h of cells in culture, indicating the disappearance of the phospholipids organized lamellae in the lamellar bodies. Similar results were obtained by using an immunocytochemical approach with antibodies raised against the apoprotein component of rat surfactant. A progressive decrease in immuno-staining corresponded to the disappearance of the lamellar bodies, and birefringence. Changes in lectins binding to the cultured type II cells were also observed. Freshly isolated and one day old cultured cells could bind Macula pomifera (MPA) but not Arachis hypogaea (PA) lectins. The reverse was found in 6-7 days old cultured cells which had the ability to bind PA but not MPA the advantage of using various cytochemical techniques for studying the development of type II cells in culture is being discussed.     

Peptide Modulators of ASIC Channels of the Sea Anemone Urticina aff. coriacea (Cuvier, 1798) from the Sea of Okhotsk

Russian Journal of Marine Biology - A search for biologically active peptides has been performed in an aqueous extract of the sea anemone Urticina aff. coriacea (Cuvier, 1798) (Actiniidae) from the...     

Clonal differences in the response of dark and light reactions of photosynthesis to elevated atmospheric CO2 in poplar

Two hybrid poplar (Populus) clones (i.e., fast growing clone Beauprè and slow growing clone Robusta) were grown for two years from cuttings at close spacings in open top chambers (OTCs) under ambient (AC) and elevated [EC = AC + 350 μmol(CO2) mol-1] CO2 treatments. For clone Beauprè no down-regulation of photosynthesis was observed. Two years of growing under EC resulted in an increase in quantum yield of photosystem 2 (PS2), steady state irradiance saturated rate of net photosynthesis (P _Nmax), chlorophyll (Chl) content, and ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPC) activity for this clone. We suppose that under non-limiting conditions of nitrogen and phosphorus content the response to EC was by building up light-harvesting complexes of PS2 and increasing photochemical efficiency of PS2. Due to a high rate of the primary reactions of photosynthesis and a high RuBPCO activity the end product of the response to EC was an increase in PNmax and a larger saccharides content. The Robusta clone showed a depression in the primary reactions of photosynthesis under EC. We found a decrease in quantum yield of PS2, Chl and phosphorus contents, and in RuBPCO activity. However, an increase in PNmax, saccharides content and Chl a/b ratio was observed. We speculate (1) that the phosphorus deficiency in combination with an increase in CO2 concentrations may lead to a potential damage of the assimilation apparatus of the primary reactions of photosynthesis and to a decrease in photochemical efficiency of PS2; (2) that the primary target of "down-regulation" takes place at PS2 for irradiances above 150 μmol m-2 s-1. This revised version was published online in September 2006 with corrections to the Cover Date.     

Histochemical studies on the distribution of acid phosphatases in neurones of sensory ganglia; light and electron microscopy

Summary Acid phosphatase activity of rat sensory ganglia was studied by light and electron microscopic histochemistry. Neurones of trigeminal and spinal ganglia showed two characteristic patterns of -glycerophosphatase. Enzymic activity in small neurones appeared as granules, i.e. lysosomes, whereas activity in large neurones appeared as a network of filaments and scattered granules. The network pattern was due to localization of the reaction product in the Golgi apparatus and in association with the rough endoplasmic reticulum. The presence of at least two acid phosphatases was suggested on account of differences in sensitivity between the enzymes in the small and large cells toward several fixatives and inhibitors.     

Effect of pathogenic cysteine mutations on FGFR3 transmembrane domain dimerization in detergents and lipid bilayers

Mutations in fibroblast growth factor receptors are known as the genetic basis of skeletal growth disorders. The mechanism of pathogenesis, as determined by mutation-induced changes in receptor structure, interactions, and function, is elusive. Here we study three pathogenic Cys mutations, associated with either thanatophoric dysplasia or achondroplasia, in the TM domain of fibroblast growth factor receptors 3 (FGFR3). We characterize the dimerization propensities of the mutant TM domains in detergents and in lipid bilayers, in the presence and absence of reducing agents, and compare them to previous measurements of wild-type. We find that the Cys mutations increase the propensity for dimerization in detergent, with the Cys370 mutant exhibiting the highest propensity for disulfide bond formation, the Cys371 mutant having an intermediate propensity, and Cys375 the lowest. Thus, disulfide bonds readily form in detergents, with efficiency that correlates with the severity of the phenotype. In lipid bilayers, however, the Cys370 mutant, which dimerizes strongly in detergent, behaves as the wild-type, suggesting that Cys370-mediated disulfide bonds do not form between the isolated TM domains in bilayers. Thus, the nature of the hydrophobic environment plays an important role in defining the structure and flexibility of transmembrane dimers. These results and previous findings from cellular studies lead us to propose a conformational flexibility mechanism of receptor stabilization as a basis for disregulated FGFR3 signaling in thanatophoric dysplasia and achondroplasia.     

Organ transplantation in Bulgaria

The transplantation program in Bulgaria started in 1968 with renal transplantations to a child and adult woman. In 1986 the first heart transplantation was performed. To date a total of 10 heart transplants have been performed, including one combined heart/lung. A liver transplantation program was launched in 2005 with a total number of 16 transplantations—7 from living donors and 9 from deceased donors. The highest transplantation activity is registered in the field of renal transplantation. During the period 1980–2006, 462 Bulgarian recipients of kidney were transplanted in Bulgaria. The ratio between transplantations from deceased and living related donors is approximately 1:0.9. Annual transplantation activity varies among the years from 1 to 12 renal transplantations p.m.p./per year. The 1- (80.7% vs. 63.1%), 5- (57.86% vs. 39.0%) and 10-year (42.65% vs. 23.62%) graft survival rates are higher for recipients of living donor kidneys compared to those of deceased donor. In 1983 a National kidney waiting list was established. Currently the number of the registered patients eligible for renal transplantation is 885. The proportion of sensitized patients in the waiting list is 20.45% and 4.34% of them are hyperimmunized. Recently HLAMatchmaker program has been implemented not only for sensitized patients but also for those with rare alleles and haplotypes. Post-transplant immunological monitoring showed a strong association between alloantibody presence and delayed graft function (Chi-square = 10.73, P < 0.001), acute rejection (Chi-square = 14.504, P < 0.001), chronic rejection (Chi-square = 12.84, P < 0.001) and graft loss (Chi-square = 20.283, P < 0.001). Based on the experience in our transplant center a strategy for improvement of long-term renal graft survival was developed and implemented.     

Identification of various testicular cell populations in pubertal and adult cockerels

Precise identification of the male germinal stem cell population is important for their practical use in programs dedicated to the integration of exogenous genetic material in testicular tissues. In the present study, our aim was to identify germinal cell populations in the testes of pubertal and adult cockerels based on the detection of the nuclear DNA content by fluorescence-activated cell sorting (FACS) and on the expression of the Dazl and Stra8 genes in single-cell suspensions of testicular tissues. Cells with a tetraploid DNA content (4c) represent a small and equal fraction of the total germinal cell population in both pubertal and adult males. In contrast, the diploid (2c) and haploid (c) subpopulations differ significantly between ages as a consequence of different degrees of sexual maturation. A specific subpopulation of testicular cells, the side-scatter subpopulation of cells, or side population (SP), was identified at the junction between the haploid and diploid cell populations. The percentage of this cell subpopulation differs significantly in pubertal and adult cockerels, accounting for 4.1% and 1.3% of the total cell population, respectively. These four testicular cell populations were also tested for the expression of Dazl and Stra8 genes known to be expressed in premeiotic cells including stem spermatogonia. Both genes were expressed in SP, whereas the expression of either Dazl or Stra8 genes was detected only in the 4c and in the 2c testicular cell subpopulations, respectively. The correlation between the cell ploidy and Dazl/Stra8 expression was the same at both male ages. We conclude that SP cells might represent a subpopulation of germinal cells enriched in stem spermatogonia, which can be of great importance for transgenesis in chicken.     

FGFR3 dimer stabilization due to a single amino acid pathogenic mutation

Mutations in the transmembrane (TM) domains of receptor tyrosine kinases (RTKs) have been implicated in the induction of pathological phenotypes. These mutations are believed to stabilize the RTK dimers, and thus promote unregulated signaling. However, the energetics behind the pathology induction has not been determined. An example of a TM domain pathogenic mutation is the Ala391-->Glu mutation in fibroblast growth factor receptor 3 (FGFR3), linked to Crouzon syndrome with acanthosis nigricans, as well as to bladder cancer. Here, we determine the free energy of dimerization of wild-type and mutant FGFR3 TM domain in lipid bilayers using F?rster resonance energy transfer, and we show that hydrogen bonding between Glu391 and the adjacent helix in the dimer is a feasible mechanism for dimer stabilization. The measured change in the free energy of dimerization due to the Ala391-->Glu pathogenic mutation is -1.3 kcal/mol, consistent with previous reports of hydrogen bond strengths in proteins. This is the first quantitative measurement of mutant RTK stabilization in a membrane environment. We show that this seemingly modest value can lead to a large increase in dimer fraction and thus profoundly affect RTK-mediated signal transduction.