Food partitioning between breeding White-tailed Kites (Elanus leucurus; Aves; Accipitridae) and Barn Owls (Tyto alba; Aves; Tytonidae) in southern Brazil.

I examined the diet of breeding White-tailed Kites (Elanus leucurus; Aves; Accipitridae) and Barn Owls (Tyto alba; Aves; Tytonidae) in an agrarian area of southern Brazil by analyzing regurgitated prey remains. The objective was to evaluate how these raptors, which differ markedly in their hunting activity periods (owls are nocturnal and kites diurnal), share their mammalian food component. 2,087 prey consumed by Barn Owls and 1,276 by White-tailed Kites were identified. They presented a high overlap of food-niches (Piankas index was 0.98). Based on the daily activity period of their main small mammal prey, a lower overlap would be expected. The crepuscular/nocturnal Mus musculus was the main prey for the diet of breeding Barn Owls (81%) and White-tailed Kites (63%). This small exotic rodent provided 63% of the small mammal biomass ingested by owls and 44% by kites. Larger native small mammals were also considered important for the diet of kites, mainly because of their biomass contribution. Although these raptors differ markedly in their hunting activity periods, Barn Owls and White-tailed Kites are very similar predators in southern Brazil, overlapping their diets.     

Mapping the integrin alpha V beta 3-ligand interface by photoaffinity cross-linking

Integrins are cell surface adhesion molecules involved in mediating cell-extracellular matrix interactions. High-resolution structural data are not available for these heterodimeric receptors. Previous cross-linking studies of integrins aimed at elucidating the nature of the receptor-ligand interface have been limited to identification of relatively large binding domains. To create reagents for "photoaffinity scanning" of the RGD-binding site of human integrin alpha V beta 3, new conformationally constrained ligands were designed. These photoreactive ligands are based on cyclo Ac-[Cys-Asn-Dmt-Arg-Gly-Asp-Cys]-OH, which displays an affinity of 50 nM for alpha V beta 3. This molecular scaffold was modified at the C-terminus by a benzophenone-containing amino acid residue, L-4-benzoylphenylalanine (Bpa). At the N-terminus, a molecular tag was introduced in the form of radioactive iodine or biotin. The newly designed tagged photoreactive RGD-containing ligands display an affinity of 0.5-0.7 microM for alpha V beta 3, and cross-link efficiently and specifically to the receptor. A 100 kDa band corresponding to the beta 3 subunit-ligand conjugate was detected as the major cross-linking product. Cross-linking was dependent upon the presence of Ca2+ and Mg2+ ions, and was competitively inhibited by a nonphotoreactive ligand. Enzymatic and chemical digestions of the radiolabeled photoconjugate enabled identification of a 20-amino acid fragment between positions 99 and 118 in the beta 3 chain of the integrin as the contact domain for ligand at a site adjacent to the C-terminal portion of the RGD triad.     

Selective recycle of viable animal cells by coupling of airlift reactor and cell settler

A new system for the perfusion culture of animal cells in suspension is described. It consists of an airlift loop reactor and a settling tank for cell retention. Insufficient nutrient and oxygen supply of the cells in the settling tank was prevented by cooling the cell suspension before entering the settler. As a result, the catabolic activity of the cells in the settler was reversibly reduced. Furthermore, the density gradient induced by cooling caused a liquid motion through the settler. Thus, it was not necessary to pump medium containing shear, sensitive cells. With this simple system, it was possible to prduce 2 to 5 g of antibodies in a 5.4-L reactor in continuous runs of 400 to 600 h. The productivity was increased by a factor of 17 and the cell density was 4 times higher in comparison with the corresponding batch system. The cell retention system was found to have the property of separating viable and nonviable cells. With the increasing perfusion rate, dead cells and debris were preferably washed out. For perfusion rates up to 1.3 d(-1), the retention efficiency of the settler was nearly 100% for viable cells; hence, this system may show advantages at the industrial scale.     

Development of optimized transfectoma cell lines for production of chimeric antibodies in hollow fiber cell culture systems

Methods for the selection of transfectoma cells that express large quantities of mouse-human chimeric antibodies have been develped. SP2/0 mouse myeloma cells were transfected with pSV2-gpt and pSV2-neo based immunoglobulin expression vectors. Double transfectants were selected using the xanthine-guanine phosphoribosyl transferase (gpt)and the neomycin (neo) selection marker genes. ELISA-based screening of transfectoma clones resulted in the isolation of IgG-producing transfectomas. Introduction of the kappa light-chain 3'-enhancer into the light-chain expression vector significantly increased immunoglobulin expression, but only when the enhancer was located at its physiological site, 9 kb downstream of the kappa constant region exon. With some of the transfectomas, final yields of up to 80 mg/L of chimeric IgG were obtained in conventional flask cultures using serum-free growth medium. A pilot-scale AcuSyst Maximizer hollow fiber cell culture system was used for the production of gram amounts of chimeric IgG. Results obtained with different transfectoma clones in conventional culture were not fully predictive for yields in the hollow fiber system. In contrast, differences in productivity between individual clones in the laboratory-scale Tecnomouse cell culture unit were comparable with those in the Maximizer system. Up to 200 mg of chimeric IgG were produced per day in one Maximizer bioreactor. (c) 1995 John Wiley & Sons, Inc.     

Lunar and temperature effects on activity of free-living desert hamsters (Phodopus roborovskii,Satunin 1903)

Time management of truly wild hamsters was investigated in their natural habitat in Alashan desert, Inner Mongolia, China during summer of 2009, 2010, and 2012. Duration of activity outside their burrows, duration of foraging walks, and nocturnal inside stays were analyzed with the aim to elucidate impact of moon, ambient, and soil temperature. Animal data were determined using radio frequency identification (RFID) technique; for that purpose, individuals were caught in the field and marked with passive transponders. Their burrows were equipped with integrated microchip readers and photosensors for the detection of movements into or out of the burrow. Lunar impact was analyzed based on moon phase (full, waning, new, and waxing moons) and moon disk size. A prolongation of aboveground activity was shown with increasing moon disk size (Spearman ρ?=?0.237; p?=?0.025) which was caused by earlier onsets (ρ?=?0.161; p?=?0.048); additionally, foraging walks took longer (Pearson r?=?0.037; p?=?0.037). Temperature of different periods of time was analyzed, i.e., mean of whole day, of the activity phase, minimum, and maximum. Moreover, this was done for the current day and the previous 3 days. Overall, increasing ambient and soil temperatures were associated with shortening of activity by earlier offsets of activity and shorter nocturnal stays inside their burrows. Most influential temperatures for activity duration were the maximum ambient temperature, 3 days before (stepwise regression analysis R?=?0.499; R ²?=?0.249; F?=?7.281; p?=?0.013) and soil temperature during activity phase, 1 day before (R?=?0.644; R ²?=?0.283; F?=?7.458; p?=?0.004).     

Effect of an adenosine A(1) receptor agonist and a novel pyrimidoindole on membrane properties and neurotransmitter release in rat cortical and hippocampal neurons

Activation of adenosine A(1) receptors by endogenous adenosine plays a neuroprotective role under various pathophysiological conditions including hypoxia. Intracellular recordings were made in rat pyramidal cells of the somatosensory cortex. Hypoxia (5 min) induced a membrane depolarization and a decrease of input resistance. The A(1) receptor agonist N(6)-cyclopentyladenosine (CPA, 100 microM) reversibly inhibited the hypoxic depolarization. The inhibition was also present after blockade of the A(2A), A(2B) and A(3) receptor subtypes by selective antagonists. CPA had no effect on the hypoxic decrease of input resistance. 1,3-Dipropyl-8-cyclopentylxanthine (DPCPX), a selective A(1) receptor antagonist, which did not alter hypoxic depolarization when given alone abolished the inhibitory effect of CPA. Neither CPA nor DPCPX influenced membrane potential or apparent input resistance under normoxic conditions. The novel pyrimidoindole (R)-9-(1-methylbenzyl)-2-(4'-pyridyl)-9H-pyrimido[4,5-b]indole-4-amine (APPPI, 1 and 10 microM) reversibly diminished hypoxic depolarization but had no significant effect on input resistance. The effect of APPPI at a concentration of 1 microM, but not at 10 microM, was blocked by DPCPX (0.1 microM). CPA (100 microM) inhibited [(3)H]-noradrenaline ([(3)H]-NA) release from rat hippocampal brain slices significantly only in the presence of rauwolscine (0.1 microM), an alpha(2)-adrenoceptor antagonist. APPPI (1 and 10 microM) exhibited an inhibitory effect similar to that observed with CPA. The effects of both CPA and APPPI were antagonized by DPCPX (0.1 microM). The present data suggest that mainly presynaptic mechanisms prevent neurons from hypoxic changes by an inhibition of transmitter release. However, in contrast to CPA, APPPI exhibited additional effects, which require further investigation.     

Habitat associations of small mammals in southern Brazil and use of regurgitated pellets of birds of prey for inventorying a local fauna.

We inventoried terrestrial small mammals in an agricultural area in southern Brazil by using trapping and prey consumed by Barn Owls (Tyto alba) and White-tailed Kites (Elanus leucurus). Small mammals were trapped in three habitat types: corn fields, uncultivated fields ("capoeiras"), and native forest fragments. A total of 1,975 small mammal specimens were trapped, another 2,062 identified from the diet of Barn Owls, and 2,066 from the diet of White-tailed Kites. Both trapping and prey in the predators' diet yielded 18 small mammal species: three marsupials (Didelphis albiventris, Gracilinanus agilis, and Monodelphis dimidiata) and 15 rodents (Akodon paranaensis, Bruceppatersonius iheringi, Calomys sp., Cavia aperea, Euryzygomatomys spinosus, Holochilus brasiliensis, Mus musculus, Necromys lasiurus, Nectomys squamipes, Oligoryzomys nigripes, Oryzomys angouya, Oxymycterus sp.1, Oxymycterus sp.2, Rattus norvegicus, and Rattus rattus (Linnaeus, 1758)). The greatest richness was found in the uncultivated habitat. We concluded that the three methods studied for inventorying small mammals (prey in the diet of Barn Owls, White-tailed Kites, and trapping) were complementary, since together, rather than separately, they produced a better picture of local richness.