5-Fluoro-4-thiouridine phosphoramidite: new synthon for introducing photoaffinity label into oligodeoxynucleotides

The synthesis of phosphoramidite of 5-fluoro-4-thio-2'-O-methyluridine is described. An appropriate set of protecting groups was optimized including the 4-thio function introduced via 4-triazolyl as the 4-(2-cyanoethyl)thio derivative, and the t-butyldimethyl silyl for 2' and 3' hydroxyl protection, enabling efficient synthesis of the phosphoramidite. These protecting groups prevented unwanted side reactions during oligonucleotide synthesis. The utility of the proposed synthetic route was proven by the preparation of several oligonucleotides via automated synthesis. Photochemical experiments confirmed the utility of the synthon.     

Cloning and Some Novel Characteristics of Mitochondrial Hsp70 from Chinese Hamster Cells

The cDNA for Chinese hamster mitochondrial Hsp70 (mHsp70) was cloned and sequenced using a polymerase chain reaction probe based on conserved regions in the Hsp70 family of proteins. The encoded protein consists of 679 amino acids which includes a N-terminal mitochondrial targeting sequence of 46 amino acids. The mHsp70 protein contains several sequence signatures that are characteristics of prokaryotic and eukaryotic organellar Hsp70 homologs. In a phylogenetic tree based on Hsp70 sequences, it branches with the gram-negative proteobacteria, supporting the endosymbiotic origin of mitochondria from this group of prokaryotes. The mHsp70 cDNA was transcribed and translatedin vitroand its import into isolated rat heart mitochondria was examined. The precursor mHsp70 was converted into a mature form of lower molecular mass (≈71 kDa) which became resistant to trypsin digestion. The import of mHsp70 into mitochondria was not observed in the presence of an uncoupler of energy metabolism or when the N-terminal presequence was lacking. The cDNA for mHsp70 was expressed inEscherichia coliand a polyclonal antibody to the purified recombinant protein was raised. The antibody shows no cross-reactivity to recombinant cytosolic Hsp70 protein and in 2-D gel blots it reacted specifically with the mHsp70 protein only. In immunofluorescence experiments, the antibody predominantly labeled mitochondria, and the observed labeling pattern was identical to that seen with a monoclonal antibody to the mitochondrial Hsp60 chaperonin. The affinity-purified antibody to mHsp70 was also employed to examine the subcellular distribution of the protein by cryoelectron microscopy and the immunogold-labeling technique. In these experiments, in addition to mitochondria, labeling with mitochondrial Hsp70 antibody was also observed on the plasma membrane and in unidentified cytoplasmic vesicles and granules. These studies raise the possibility that similar to the Hsp60 chaperonin and a number of other mitochondrial proteins, mHsp70 may have an extramitochondrial role.     

Selfish long-term benefits of hoarding in the Siberian jay

We followed the consumption of cached food in Siberian jay flocksduring winter. Siberian jay flocks contain kin associationscomposed of parents in company with mature offspring. Further,flocks often contain immigrants not closely related to the groupmembers. We studied the extent to which Siberian jay offspringkept caches to themselves or whether they showed a kin biasand shared cached food mutualistically with relatives (parents/siblings)thus making an inclusive fitness gain. We recorded the retrievalof cached food using radio-ptilochronology, a technique thatdetects the consumption of a radioactively labeled food itemin the growth bar laid down in a growing feather the day ofconsumption. Food caching entailed a selfish benefit to thehoarder. The hoarding bird showed a substantial "recovery advantage"and retrieved its own cached food seven times as often as anyof its flock mates. There was no evidence for mutualistic sharingof caches among relatives. This selfish benefit persisted throughoutthe 7-week period for which we could follow the consumptionof labeled food.     

Pyrola chlorantha Sw.—eine beachtenswerte Art des Osthimalaya

Zusammenfassung Aus dem zentralen Bhutan wird ein neues Taxon beschrieben:Pyrola chlorantha Sw. subsp.fallax Kísa (SektionChlorantha Kísa, SeriesChloranthae Kísa). Die nächsten Fundorte der disjunktnordhemi-sphärisch verbreitetenP. chlorantha subsp.chlorantha liegen im mongolisch-sowjetischen Grenzgebiet (Baikal-Dahurien)     

Differentially measuring infrared analyzer with an air-conditioned exposure chamber for photosynthetic rate measurements

Je popsáno laboratorní gazometrické za?ízení pro stanovení intensity fotosynthesy v regulovaných podmínkách, umo?ňující difereněním zapojením infra?erveného analysátoru dosáhnout vysoké p?esnosti p?i stanovení rozdílu v koncentraci CO₂ mezi kontrolnín vzduchem a vzduchem ochuzeným o asimilovaný CO₂. Infra?ervený analysátor je zapojen tak, ?e jednou trubicí trvale prochází vzduch kontrolní a druhou “ochuzený” vzduch pokusný, tak?e výchylka registrujícího p?ístroje, kterou je mo?no zna?ně zesílit, odpovídá p?ímo absolutní hodnotě ochuzení, tedy p?i známém pr?toku p?ímo intensitě fotosynthesy. Tak je pr?měrná chyba stanovení intensity fotosynthesy vyjád?ena chybou stanovení koncentrace CO₂ p?ístrojem p?i pou?itém zesílení v procentech diference a nikoliv v procentech několikánásobně vy??í absolutní koncentrace CO₂, jak je tomu p?i absolutním zapojení. P?ípadné kolísání koncentrace CO₂ v kontrolním vzduchu je registrováno druhým infra?erveným analysátorem, který je zapojen normálně, tj. absolutně. K exposici rostliny běhern pokusu je pou?íváno jednoduché klimatisované komory s regulovanou teplotou (±0,2° C) a vlhkostí (±2 a? 5% rel. vlhkosti) vzduchu s vysokou regulovancu stabilní intensitou osvětlení (a? do cca 3,2. 10⁶ erg. cm^?2. sec^?1) a s mo?ností zvy?ovat koncentraci CO₂ vevzduchu.     

Response of brain amino acid metabolism to ketosis

Our objective was to study brain amino acid metabolism in response to ketosis. The underlying hypothesis is that ketosis is associated with a fundamental change of brain amino acid handling and that this alteration is a factor in the anti-epileptic effect of the ketogenic diet. Specifically, we hypothesize that brain converts ketone bodies to acetyl-CoA and that this results in increased flux through the citrate synthetase reaction. As a result, oxaloacetate is consumed and is less available to the aspartate aminotransferase reaction; therefore, less glutamate is converted to aspartate and relatively more glutamate becomes available to the glutamine synthetase and glutamate decarboxylase reactions. We found in a mouse model of ketosis that the concentration of forebrain aspartate was diminished but the concentration of acetyl-CoA was increased. Studies of the incorporation of 13C into glutamate and glutamine with either [1-(13)C]glucose or [2-(13)C]acetate as precursor showed that ketotic brain metabolized relatively less glucose and relatively more acetate. When the ketotic mice were administered both acetate and a nitrogen donor, such as alanine or leucine, they manifested an increased forebrain concentration of glutamine and GABA. These findings supported the hypothesis that in ketosis there is greater production of acetyl-CoA and a consequent alteration in the equilibrium of the aspartate aminotransferase reaction that results in diminished aspartate production and potentially enhanced synthesis of glutamine and GABA.