Effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and leptin on hypothalamic mRNA expression of factors participating in food intake regulation in a TCDD-sensitive and a TCDD-resistant rat strain

An acutely toxic dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) leads to a drastically and permanently reduced feed intake and wasting by an unknown mechanism. We focused on the possible interference of TCDD with hypothalamic factors known to take part in the regulation of eating and metabolism, utilizing the over 1000-fold TCDD-sensitivity difference between Long-Evans (Turku/AB; L-E) and Han/Wistar (Kuopio) rats. The mRNA expression of 18 hypothalamic factors (including NPY, AgRP, and CART) was measured by quantitative RT-PCR at 6, 24 and 96 h after TCDD administration. The effects of TCDD were compared with those of leptin and with feed restriction employing a TCDD dose that elicited a severe reduction of feed intake in L-E rats. TCDD mainly modified expression of orexigenic factors causing an initial suppression followed by reversal to enhanced expression by 96 h. The latter was also seen in feed-restricted controls. In contrast, leptin altered both orexigenic and anorexigenic factor mRNAs in a more even manner and its effects were clustered at 6 h. The transient nature of feeding-promoting factor suppression does not strongly support a key role for this phenomenon in TCDD-induced wasting syndrome. However, the fact that TCDD mainly affected orexigenic factors and the temporal differences in response found between the rat strains warrant further research.     

Production of pectin methylesterase from Erwinia chrysanthemi B374 in Bacillus subtilis

Summary The gene coding for pectin methylesterase (PME) of Erwinia chrysanthemi B374 (pme) was cloned by a polymerase chain reaction. The pme gene was expressed in Bacillus subtilis using a secretion vector based on the promoter and signal sequence of the -amylase gene from B. amyloliquefaciens. The cultivation of B. subtilis cells carrying the cloned pme resulted in efficient secretion of PME into the culture medium based on enzymatic and sodium dodecyl sulphate-polyacrylamide gel electrophoresis characterizations. The NH₂-terminal sequence analysis of the secreted PME revealed two different NH₂-termini. Heterologous processing was probably due to a second putative signal peptidase cleavage site at the joint region between the PME and -amylase signal peptide. Offprint requests to: R. Heikinheimo     

Climate change affects populations of northern birds in boreal protected areas

Human land-use effects on species populations are minimized in protected areas and population changes can thus be more directly linked with changes in climate. In this study, bird population changes in 96 protected areas in Finland were compared using quantitative bird census data, between two time slices, 1981-1999 and 2000-2009, with the mean time span being 14 years. Bird species were categorized by distribution pattern and migratory strategy. Our results showed that northern bird species had declined by 21 per cent and southern species increased by 29 per cent in boreal protected areas during the study period, alongside a clear rise (0.7-0.8 °C) in mean temperatures. Distribution pattern was the main factor, with migratory strategy interacting in explaining population changes in boreal birds. Migration strategy interacted with distribution pattern so that, among northern birds, densities of both migratory and resident species declined, whereas among southern birds they both increased. The observed decline of northern species and increase in southern species are in line with the predictions of range shifts of these species groups under a warming climate, and suggest that the population dynamics of birds are already changing in natural boreal habitats in association with changing climate.     

Role of ω-Conotoxin GVIA-Sensitive Ca2+ Entry in Angiotensin II-Stimulated [3H]Phorbol 12, 13-Dibutyrate Binding in Bovine Adrenal Medullary Cells

Abstract: The relative contributions of Ca²⁺ influx and intracellular Ca²⁺ mobilization were examined for angiotensin II-stimulated [³H]phorbol 12, 13-dibutyrate binding, which reflects the level of activated protein kinase C in bovine chromaffin cells. Angiotensin II receptors activate phospholipase C in chromaffin cells, leading to a shortlived mobilization of intracellular Ca²⁺. Angiotensin II-stimulated [³H]phorbol 12, 13-dibutyrate binding was largely blocked in Ca²⁺-free buffer and by pretreatment with the Ca²⁺-channel blocker ω-conotoxin GVIA. The [³H]phorbol 12, 13-dibutyrate binding response to [Sar¹]angiotensin II also appeared to be voltage sensitive, as no additivity was observed with the response to the depolarizing agent 4-aminopyridine (3 m M ). Threshold sensitivities of the extra-and intracellular Ca²⁺-mobilizing pathways to angiotensin II were similar, and all examined effects of angiotensin II in these cells were apparently mediated by losartan-sensitive (AT₁-Iike) receptors. The dependence of angiotensin II-stimulated [³H]phorbol 12, 13-dibutyrate binding on extracellular Ca²⁺ entry, in contrast to stimulation by other phospholipase C-linked receptor agonists (bradykinin and methacholine), suggests that angiotensin II preferentially stimulates protein kinase C translocation to the plasma membrane, rather than to internal membranes, in bovine adrenal medullary cells.     

Accumulation of altered aspartyl residues in erythrocyte membrane proteins from patients with sporadic amyotrophic lateral sclerosis

Spontaneous protein deamidation of labile asparagines (Asn), generating abnormal l-isoaspartyl residues (IsoAsp), is associated with cell aging and enhanced by an oxidative microenvironment. The presence of isopeptide bonds impairs protein structure/function. To minimize the damage, IsoAsp can be “repaired” by the protein l-isoaspartyl/d-aspartyl O-methyltransferase (PIMT) and S-adenosylmethionine (AdoMet) is the methyl donor of this reaction. PIMT is a repair enzyme that initiates the conversion of l-isoAsp (or d-Asp) residues to l-Asp residues. Amyotrophic lateral sclerosis (ALS) is a severe neurodegenerative disease principally affecting motor neurons. The condition of oxidative stress reported in familial and sporadic forms of ALS prompted us to investigate Asn deamidation in ALS tissue. Erythrocytes (RBCs) were selected as a model system since they are unable to replace damaged proteins and protein methylesterification is virtually the only AdoMet-consuming reaction operating in these cells. Our data show that, in vitro assay, abnormal IsoAsp residues were significantly higher in ALS patients erythrocyte membrane proteins with an increased methyl accepting capability relative to controls (p < 0.05). Moreover, we observed a reduction in AdoMet levels, while AdoHcy concentration was comparable to that detected in the control, resulting in a lower [AdoMet]/[AdoHcy] ratio. Then, the accumulation of altered aspartyl residues in ALS patients is probably related to a reduced efficiency of the S-adenosylmethionine (AdoMet)-dependent repair system causing increased protein instability at Asn sites. The increase of abnormal residues represents a new protein alteration that may be present not only in red blood cells but also in other cell types of patients suffering from ALS.     

Spatial Variation of Bird Species in Landscapes Dominated by Old-growth Forests in Northern Boreal Finland

An important issue in conservation planning is to study the distribution and abundance patterns of species in natural landscapes. Information of concordant and clumped species distribution pattertns can enhance attempts to focus conservation efforts in sites/areas preferred by various species. We compared bird species abundance, community composition and species distribution patterns in three large old-growth forest areas (size 40–120 km²) in northern boreal Finland on the basis of quantitative bird censuses. Total bird density and species composition, and mean density and variances of the most abundant species were highly similar between the three areas. Most of the bird species were distributed randomly within the areas and species preferring old-growth forests showed compensatory density variation without preference for any sub-area. Mean density of species preferring old-growth forests was significantly higher in the studied large old-growth forests than in smaller old-growth forests and was about threefold higher than regional density of species in a predominantly managed landscape matrix. Bird species preferring old-growth forests are probably not dependent on a certain specific part of a virgin boreal forest landscape but rather on the overall size of the high-quality and diverse old-growth forest area.     

Recent sedimentological conditions in the delta of the river Kyrönjoki,Western Finland

The estuary of the River Kyrönjoki was echo sounded and, on the basis of these results the delta was delimited and areas of erosion and accumulation were separated. Samples of the sediment surface (0–2 cm) from 65 sites in the estuary were analysed for organic content, C, N, P, Ca, Fe, Mn, Pb, Cu, Zn, Cd and Hg. On the basis of the chemical analyses the delta was divided into two parts according to the sedimentary conditions: (I) In the inner part, where acid fresh water of c. pH 5.5 from the river is predominant, the conditions were reducing. The Fe/Mn ratio of the sediment was high. The heavy metal content was usually low. The Hg content in the organic compounds of the sediment was, due to the acidity, higher than in the outer part. (II) In the outer part of the delta, where neutral brackish water of pH c. 7.0 dominates, the sedimentary conditions were mostly oxidizing. The Fe/Mn ratio was very low. The heavy metal and organic content was mostly high in the accumulation areas. This is because the fine-grained allochthonous material, which has a high heavy metal and organic content, flocculates and precipitates when it reaches the brackish water of the Gulf of Bothnia, and the salinity and the pH are higher.     

Island selection on mammalian life-histories: genetic differentiation in offspring size

Background  

Since Darwin's pioneering work, evolutionary changes in isolated island populations of vertebrates have continued to provide the strongest evidence for the theory of natural selection. Besides macro-evolutionary changes, micro-evolutionary changes and the relative importance of natural selection vs. genetic drift are under intense investigation. Our study focuses on the genetic differentiation in morphological and life-history traits in insular populations of a small mammal the bank vole Myodes glareolus.     

Plasmatic exosomes from prostate cancer patients show increased carbonic anhydrase IX expression and activity and low pH

Abstract

Acidity, hypoxia and increased release of exosomes are severe phenotypes of tumours. The regulation of pH in tumours involves the interaction of several proteins, including the carbonic anhydrases which catalyze the formation of bicarbonate and protons from carbon dioxide and water. Among CA isoforms, CA IX is over-expressed in a large number of solid tumours, conferring to cancer cells a survival advantage in hypoxic and acidic microenvironment, but there isn’t evidence that CA IX expression could have a real clinical impact. Therefore, in this study for the first time the expression and activity of CA IX have been investigated in the plasmatic exosomes obtained from patients with prostate carcinoma (PCa). For this purpose, the study was performed through different methodological approaches, such as NTA, western blot analysis, enzyme activity assay, Nanoscale flow cytometry, ELISA, confocal microscopy. The results showed that PCa exosomes significantly overexpressed CA IX levels and related activity as compared to healthy donors. Furthermore, CA IX expression and activity were correlated to the exosome intraluminal pH, demonstrating for the first time that PCa exosomes are acidic. Our data suggest the possible use of the exosomal CA IX expression and activity as a biomarker of cancer progression in PCa.