Fever produced in rabbits by N6, O2′-dibutyryl adenosine 3′, 5′-cyclic monophosphate

Zusammenfassung Die Injektion von N⁶-2-O-Dibutyryl-adenosin-3 5-monophosphat in den lateralen Hirnventrikel von Kaninchen führt zu einer langanhaltenden Fieberreaktion, was vermuten lässt, dass zyklisches Adenosin-35-monophosphat als weiterer Mediator in der Genese des Fiebers fungiert.     

Synthesis and application of the photolabile guanosine 3′,5′-phosphoric-o-nitrobenzylester

Summary The o-nitrobenzylester of guanosine 3,5-phosphate is prepared and the photolysis studied. This photoactive form of cGMP should be an agent capable of altering the intracellular concentration of cGMP through membrane penetration and subsequent photolysis.Acknowledgments. We would like to thank the Deutsche Forschungsgemeinschaft and the Universität Konstanz for kindly supporting this research.     

Correlation between 3′,5′, c-AMP levels and thyrotropin in separated rat pituitary thyrotropic cells

Summary The correlation between 3,5, c-AMP levels, TSH content and secretion of separated thyrotropic cells was studied. Incubation of the separated cells with 1, 10 and 100 ng of TRH does not change the 3,5, c-AMP levels, despite the significant rises of the TSH level. Dibutyryl c-AMP causes rise in TSH content, with no indication of its secretion. PGE₂ 10^–5 increased 3, 5, c-AMP levels with no change in the content or secretion of TSH in separated thyrotropic cells.     

Adenosine 3′,5′-cyclic monophosphate levels in maize roots

Summary Endogenous levels of adenosine 3,5-cyclic monophosphate (cAMP) in maize (cv. LG 11) root cells, grown in light and dark conditions, were found to be 309 and 387 pmoles/g of fresh tissue respectively.This investigation was partially supported by USA National Science Foundation grant No. CDP 8004200 to the senior author.     

Flow-induced release of adenosine 5′-triphosphate from endothelial cells of the rat mesenteric arterial bed

Adenosine 5-triphosphate (ATP) was released into the perfusate of rat isolated mesenteric arterial beds during each of two consecutive increases in flow. There was no significant difference between the amounts of ATP released on each occasion. Substance P was also released into the perfusate by increased flow, although its release was more variable. Removal of the endothelium of the mesenteric vessels with sodium deoxycholate led to a significant reduction (74%) in the amount of ATP released compared with the release before the endothelium had been removed. This suggests that the ATP released into the mesenteric arterial perfusate during increased flow arises from endothelial cells.     

Reaction of pyridoxal-5′-phosphate with γ-carboxyglutamic acid

Summary Pyridoxal-5-phosphate (PLP) reacts with -carboxyglutamic acid (Gla) to form a stable complex absorbing at 325 nm. It is suggested that a condensation occurs in which the formyl group of PLP reacts with the -amino group and the carbon atom of Gla to give a pyrrolidine derivative.     

Glucose-evoked recovery of hepatic thyroxine 5′-deiodinase independent of de novo protein synthesis in fasted rat

Summary The glucose-evoked recovery of Type I thyroxine 5-deiodinase activity in the hepatic microsomes of fasted rat was not inhibited by either cycloheximide, puromycin or actinomycin D during 3 h after glucose feeding; however, [³H]-leucine uptake by the liver or the hepatic microsomal fraction was significantly inhibited by cycloheximide and puromycin but not by actinomycin D. These results indicate that the glucose-evoked recovery of deiodinase activity may be independent of de novo protein synthesis.     

Crosstalk of protein kinase C ε with Smad2/3 promotes tumor cell proliferation in prostate cancer cells by enhancing aerobic glycolysis

Protein kinase C ε (PKCε) has emerged as an oncogenic protein kinase and plays important roles in cancer cell survival, proliferation, and invasion. It is, however, still unknown whether PKCε affects cell proliferation via glucose metabolism in cancer cells. Here we report a novel function of PKCε that provides growth advantages for cancer cells by enhancing tumor cells glycolysis. We found that either PKCε or Smad2/3 promoted aerobic glycolysis, expression of the glycolytic genes encoding HIF-1α, HKII, PFKP and MCT4, and tumor cell proliferation, while overexpression of PKCε or Smad3 enhanced aerobic glycolysis and cell proliferation in a protein kinase D- or TGF-β-independent manner in PC-3M and DU145 prostate cancer cells. The effects of PKCε silencing were reversed by ectopic expression of Smad3. PKCε or Smad3 ectopic expression-induced increase in cell growth was antagonized by inhibition of lactate transportation. Furthermore, interaction of endogenous PKCε with Smad2/3 was primarily responsible for phosphorylation of Ser213 in the Samd3 linker region, and resulted in Smad3 binding to the promoter of the glycolytic genes, thereby promoting cell proliferation. Forced expression of mutant Smad3 (S213A) attenuated PKCε-stimulated protein overexpression of the glycolytic genes. Thus, our results demonstrate a novel PKCε function that promotes cell growth in prostate cancer cells by increasing aerobic glycolysis through crosstalk between PKCε and Smad2/3.     

Antiproliferative effects of 4′, 5′-unsaturated adenine nucleosides on leukemia L 1210 cells in vitro

Summary Several 4, 5-unsaturated adenine nucleosides were shown to have antiproliferative activity against L 1210 leukemia cells in vitro. The active nucleosides were cytotoxic to the L 1210 cells as demonstrated by Trypan Blue uptake. The cytotoxicity was not induced by alterations in the ribonucleoside and deoxyribonucleoside triphosphate levels of the L 1210 cells.     

Rosiglitazone counteracts palmitate-induced β-cell dysfunction by suppression of MAP kinase, inducible nitric oxide synthase and caspase 3 activities

Chronic exposure of pancreatic islets to elevated levels of palmitate leads to beta-cell dysfunction. We examined possible involvement of mitogenactivated protein kinases (MAPKs) and caspase-3 in palmitate-induced beta-cell dysfunction and tested the influence of the anti-diabetic drug rosiglitazone (ROZ). Palmitate amplified glucose-stimulated augmentation of intracellular free calcium ([Ca2+]i) and insulin secretion in incubated islets. ROZ suppressed this amplification, whereas it modestly augmented glucose-induced increase in these events. ROZ suppressed short-term palmitate-induced phosphorylation of pro-apoptotic MAPKs, i.e., SAPK/JNK and p38. Long-term islet culturing with palmitate induced inducible nitric oxide synthase (iNOS) and activated SAPK/JNK-p38. ROZ counteracted these effects. Both palmitate and cytokines activated caspase-3 in MIN6c4-cells and isolated islets. ROZ suppressed palmitate- but not cytokine-induced caspase-3 activation. Finally, after palmitate culturing, ROZ reversed the inhibitory effect on glucose-stimulated insulin release. We suggest that ROZ counteracts palmitateinduced deleterious effects on beta-cell function via suppression of iNOS, pro-apoptotic MAPKs and caspase-3 activities, as evidenced by restoration of glucose-stimulated insulin release.     

Effect of spike disease on the 5′-and 3′-nucleotidase activities in sandal plants

Summary In the sandal plant affected by spike disease, caused by mycoplasma-like organisms, the changes occurring in the 5-and 3-nucleotidase activities were studied and their possible significance discussed.     

The effect of cyclic cytidine 3′,5′-monophosphate (cCMP) on the in vitro development,hatching and attachment of the mouse blastocyst

Summary The in vitro development and attachment of hatched mouse blastocysts on the untreated substratum was enhanced by 10 M dibutyryl cCMP (dbcCMP). The result suggests that cCMP has an effect on embryonic development and on the blastocyst attachment process.25 November1986     

KIF5C: A new binding partner for protein kinase CK2 with a preference for the CK2α’ subunit

Protein kinase CK2 is a highly conserved serine/threonine kinase that is ubiquitously expressed in eukaryotic cells. CK2 is a constitutively active tetrameric enzyme composed of two catalytic α and/or α’-subunits and two regulatory β-subunits. There is increasing evidence that the individual subunits may have independent functions and that they are asymmetrically distributed inside the cell. To gain a better understanding of the functions of the individual subunits, we employed a yeast-two-hybrid screen with CK2α and CK2α’. We identified the motor neuron protein KIF5C as a new binding partner for CK2. The interaction found in the yeast-two-hybrid screen was confirmed by co-sedimentation analysis on a sucrose density gradient and by co-immunoprecipitation analysis. Pull-down experiments and surface plasmon resonance spectrometry revealed a direct binding of KIF5C to CK2α’. Co-localization studies with neuroblastoma cells, bone marrow and with primary neurons confirmed the biochemical analysis that KIF5C preferentially bound to CK2α’. Received 8 August 2008; received after revision 3 November 2008; accepted 4 November 2008     

IgG purification to measure the level of an iodinated thyroglobulin peptide,the 3,5,3′,5′ tetraiodo-l-tyrosyl-l-tyrosine in human serum

Summary Antibodies reacting with 3,5,3,5 tetraiodo-l-tyrosyl-l-tyrosine (I₂Tyr-I₂Tyr) were elicited in rabbits by immunization with an oxidized yeast conjugate coupled with I₂Tyr-I₂Tyr. Ion-exchange chromatography was used to purify immunoglobulins, in order to improve the specificity in measurement of I₂Tyr-I₂Tyr level in patient serum. IgG binding capacity versus I₂Tyr-I₂Tyr was considerably increased after immunoglobulin purification.Acknowledgments. The authors with to thank Mrs M. Ollier for her valuable technical assistance.     

Preferential hydrophobic interactions are responsible for a preference of D-amino acids in the aminoacylation of 5′-AMP with hydrophobic amino acids

We have studied the chemistry of aminoacyl AMP to model reactions at the 3 terminus of aminoacyl tRNA for the purpose of understanding the origin of protein synthesis. The present studies relate to the D, L preference in the esterification of 5-AMP. All N-acetyl amino acids we studied showed faster reaction of the D-isomer, with a generally decreasing preference for D-isomer as the hydrophobicity of the amino acid decreased. The -branched amino acids, Ile and Val, showed an extreme preference for D-isomer. Ac-Leu, the -branched amino acid, showed a slightly low D/L ratio relative to its hydrophobicity. The molecular basis for these preferences for D-isomer is understandable in the light of our previous studies and seems to be due to preferential hydrophobic interaction of the D-isomer with ademine. The preference for hydrophobic D-amino acids can be decreased by addition of an organic solvent to the reaction medium. Conversely, peptidylation with Ac-PhePhe shows a preference for the LL isomer over the DD isomer.     

5′-Nucleotidase activity in liver homogenates of rats treated with CCl4, colchicine,cycloheximide, emetine,ethanol, ethionine and 5-fluorotryptophan

Summary 5-Nucleotidase activity, an enzyme marker of the plasma membranes, increases in female rat liver homogenates following ethionine administration, while homogenates from males show no changes. Treatment with CCl₄, colchicine, cycloheximide, emetine, ethanol and 5-fluorotryptophan does not significantly modify the 5-nucleotidase activity of liver homogenates of either female or male rats.     

Antagonism by haloperidol of the suppression of exploratory locomotor activity induced by the local application of(−)3-(3-hydroxyphenyl)-N-n-propylpiperidine into the nucleus accumbens of the rat

Summary The injection of (–)3-PPP into the nucleus accumbens, 10 g/side, produced a suppression of exploratory locomotor activity without affecting treadmill locomotion. Furthermore, the suppression of exploratory locomotor activity produced by (–)3-PPP was antagonized by the administration of haloperidol, 25–50 g/kg i.p.Acknowledgments. The figures were prepared by M. Kröning at the Department of Psychology, University of Göteborg. This work was supported by Torsten and Ragnar Söderberg's Foundation, Magn. Bergvall Foundation and Åke Wiberg Foundation.