ß-eye: A benchtop system for in vivo molecular screening of labeled compounds.

Preclinical nuclear molecular imaging speeds up the mean time from synthesis to market, in drug development process. Commercial imaging systems have in general high cost, require high-cost service contracts, special facilities and trained staff. In the current work, we present ß-eye, a benchtop system for in vivo molecular screening of labeled compounds with Positron Emission Tomography (PET) isotopes. The developed system is based on a dual-head geometry, offering simplicity and decreased cost. The goal of the design is to provide 2D, real-time radionuclide images of mice, allowing the recording of fast frames and thus perform fast kinetic studies, with spatial resolution of ∼2 mm. Performance evaluation demonstrates the ability of ß-eye to provide quantitative results for injected activities lower than 1.5 MBq, which is adequate for pharmacodynamic studies in small mice.

Song perception among incipient species as a mechanism for reproductive isolation.

Many functions in behavioural processes of small passerines are regulated via vocal messages. Song plays an important role in the development of reproductive barriers and thus playback experiments can often be used for investigating the potential for reproductive isolation through behavioural mechanisms. Moltoni's warbler Sylvia (cantillans) moltonii is characterized by diagnostic vocalizations and a peculiar pattern of distribution, being parapatric and partly sympatric with the nominate Sylvia c. cantillans. With this work, we test whether these two closely related taxa react equally to their own song and to the song of the other taxon, shedding light on whether they perceive each other's songs as coming from the same species. We carried out 184 playback experiments within the mainland range of the two forms. We judged the response of the bird on a scale of scores. Each taxon responded more strongly to playback when faced with the song of its own taxon. This held true when applied only to males or females. Additionally, birds tested for both songs showed a stronger response to the song of their own taxon. The distributional context (sympatry vs. allopatry) did not affect bird response. Results indicate that a certain degree of reproductive isolation between the two taxa (because of diverged mate recognition systems) already exists; consistently with genetic data and with the peculiar pattern of distribution; this suggests that the two taxa have reached species status.

Important hints in behavioural teratology of rodents.

The paper deals with the most important items regarding the improvement of quality of experimental procedures when testing drug effects on behaviour and development of commonly-used rodent species. Current-used test procedures for immature and adult rodents exposed early developmentally are briefly described and recent advances and difficulties in their hands-on interpretation are highlighted. Comparability of measures in human and animals for drug-effect assessment is also shortly discussed. It is then stressed that studies on rodents carried out in seminaturalistic and naturalistic settings may offer a highly profitable direction for future research in behavioural teratology and toxicology. A final paragraph is dedicated to the bioethical aspects arisen from the use of large number of rodents subjects in behavioural testing.

Prenatal AZT or 3TC and mouse development of locomotor activity and hot-plate responding upon administration of the GABA(A) receptor agonist muscimol.

RATIONALE: Zidovudine (AZT) and lamivudine (3TC) are nucleoside analogues administered prenatally in clinical practice, separately or in combination, as antiretroviral drugs to prevent HIV mother-to-child transmission by inhibiting viral reverse transcriptase. In animal studies pre- and/or perinatal exposure to AZT and 3TC induce age- and sex-dependent neurobehavioural alterations in the offspring. OBJECTIVE: Investigation of short- and medium-term effects of in utero exposure to AZT or 3TC on development of the GABAergic system. METHODS: Pregnant CD-1 mice were given orally twice daily AZT (160 mg/kg), 3TC (500 mg/kg) or vehicle solution (NaCl 0.9%) from pregnancy day 10 to delivery. Offspring locomotion and nociceptive sensitivity were examined on postnatal day (pnd) 8, 14, and 28 after administration of two doses of GABAergic agonist muscimol (pnd 8 and 14: 0.05 and 0.2 mg/kg; pnd 28: 0.2 and 1.0 mg/kg). A 30-min locomotor activity test and a 60 s hot-plate test (50+/-1 degrees C) were used. RESULTS: AZT and 3TC treated mice showed a mild increase of locomotor activity after administration of the high dose muscimol on pnd 8. On pnd 14 the low muscimol dose enhanced locomotor activity in vehicle and 3TC, but not in AZT pups, whereas no prenatal treatment effect was evident on pnd 28. AZT increased nociceptive sensitivity at all ages considered. CONCLUSIONS: Prenatal AZT effects on locomotor activity appear clearly detectable after GABAergic challenge and seem to be transient. AZT effects on pain sensitivity did not appear to be dependent on GABA regulated nociceptive mechanisms. Prenatal 3TC exposure had rather limited effects on locomotor activity development, and no effect on nociception.

Limited changes in handedness and morphine reactivity in CD-1 mice after pre- and postnatal ozone exposure.

Outbred CD 1 mice were either not exposed (control group) or exposed to ozone (O3) (0.3, 0.6, or 0.9 ppm), during foetal and neonatal life until the time of weaning (postnatal day (PND) 26). On PND 70 the subjects were tested for handedness using a paw preference task assessing both the animals' capability to reach a food pellet in a feeding tube and the individual preference for the use of one of the other forepaw. O3 exposure did not affect the animals' capability to learn the task but caused changes in handedness. Specifically, females exposed to the intermediate O3 concentration showed a reduced preference for the right paw than both their same-sex controls and 0.6 ppm males. On PND 100, mice underwent a hot plate test after IP treatment by either saline or morphine HCl (10 mg/kg). The results were generally in the direction of reduced drug sensitivity after exposure to the highest concentration. The evidence for this effect was more robust in the case of an organised avoidance response (wall-rearing) than in the case of a reflexive response (limb withdrawal); in the case of the former, latency data showed an effect on both males and females while frequency data showed an effect only in females. Overall, the O3 effects are suggestive of subtle CNS changes affecting mouse behavioural responses.

Systemic delivery of a breast cancer-detecting adenovirus using targeted microbubbles.

One of the major limitations of cancer gene therapy using recombinant human adenovirus (Ad) is rapid Ad inactivation from systemic delivery. To eliminate this, biotin-coated ultrasound contrast agents, or microbubbles (MBs), were streptavidin-coupled with biotinylated antibodies to three distinct tumor vasculature-associated receptors (α(V)ß(3) integrin, P-selectin and vascular endothelial growth factor receptor-2) for systemic targeting of a previously generated vector Ad5/3-Id1-SEAP-Id1-mCherry. This cancer-specific, dual-reporter vector was loaded in the targeted MBs and confirmed by confocal microscopy. MB loading capacity was estimated by functional assays as 4.72 ± 0.2 plaque forming unit (PFU) per MB. Non-loaded (free) Ad particles were effectively inactivated by treatment with human complement. The Ad-loaded, targeted-MBs were injected systemically in mice bearing MDA-MB-231 tumors (Grp 1) and compared with two control groups: Ad-loaded, non-targeted MBs (Grp 2) and free Ad (Grp 3) administered under the same conditions. Two days after administration the blood levels of secreted embryonic alkaline phosphatase (SEAP) reporter in Grp 1 mice (16.1 ng ml(-1) ± 2.5) were significantly higher (P<0.05) than those in Grp 2 (9.75 ng ml(-1) ± 1.5) or Grp 3 (4.26 ng ml(-1) ± 2.5) animals. The targeted Ad delivery was also confirmed by fluorescence imaging. Thus, Ad delivery by targeted MBs holds potential as a safe and effective system for systemic Ad delivery for the purpose of cancer screening.

Value to wildlife of urban-agricultural parks: a case study from Rome urban area.

Urban-agricultural parks could have some advantages to wildlife because of less intensive agricultural procedures, absence of hunting pressure, and reduced human disturbance. In this study, the breeding and wintering bird communities and the small mammal community in an urban-agricultural park of Rome were compared to those of a close urban park and a close agricultural area just outside the city. The aim was to assess the best destination and management of wildlife in natural areas at the urban-rural interface. Richness and diversity of bird communities were higher in the urban-agricultural park. Due to habitat features and probably human disturbances, but not to urbanization, predation, and competition factors, the urban park drastically reduces the abundance of decreasing open-land bird species. Abundance of these species was not significantly different in the urban-agricultural park and in the agricultural area. In the urban-agricultural and urban park, bird and mammal pest species were more abundant than they were in the agricultural area. Regarding decreasing abundance of small mammal species, no significant difference among the study areas was observed. Urban-agricultural park is a better choice than urban park for wildlife. Thus, a higher number of preserved urban natural areas should be devoted to urban-agricultural parks. However, to increase the abundance of open-land species and in general wildlife, a less intensive management of cultivated and pasture patches is necessary.

Animacy effects on the production of object-dislocated descriptions by Catalan-speaking children.

This paper presents an experiment that examined two related questions. First, it examined the effects of animacy on the production of different syntactic structures and word orders by Catalan-speaking children. Secondly, it explored the relationship between age and the production of different syntactic structures by these children. The results of a picture description experiment run with eighty Catalan-speaking children aged 4;11 to 11;11 show that participants tended to produce more object-dislocated descriptions when the patient was animate than when the patient was inanimate. The results also show that while the dislocation of object clauses is a construction already consolidated at 5;0, the passive structure is a construction still not fully acquired at 11;0. A comparison between the results obtained in the present experiment with existing results from similar experiments with English-speaking children shows that there is a cross-language difference in the age at which children start producing passive clauses. We argue that frequency of exposure to a particular syntactic structure is an important factor that contributes to the acquisition of that syntactic structure. We also suggest that the effects of animacy on the production of object-dislocated descriptions can be explained by means of conceptual and lexical accessibility.

Prolonged exposure to low doses of ozone: short- and long-term changes in behavioral performance in mice.

Two separate experiments were designed to assess the effects of ozone exposure on outbred CD-1 mice. In the first experiment, adult males were exposed continuously to O3 at 0, 0.3, or 0.6 ppm for 30 days and their behavior was assessed in a 5-min open-field test on exposure days 4 and 19 and on day 3 after the end of the exposure phase. In addition, mice performed a Morris water maze task from exposure day 24 to 28. In the second experiment, adult females were exposed from 30 days prior to the formation of breeding pairs until gestational day 17 to the same doses used in the first experiment. Litters were fostered at birth to untreated dams and neurobehavioral development of the offspring was investigated until adulthood. Specifically, somatic and sensorimotor development [postnatal day (PND) 2-20], homing performance (PND 12), motor activity (PND 21), passive avoidance (PND 22-23), water maze performances (PND 70-74), and response to a nociceptive stimulus (PND 100) were assessed. Results from both experiments confirm that exposure to O3 slightly but selectively affected neurobehavioral performance in rodents. Exposure to O3 did not grossly affect neurobehavioral development, whereas it consistently impaired reversal learning in the Morris water maze test in both prenatally and adult exposed mice. Moreover, longer latency to step-through in the first trial of the passive avoidance test and a decrease in wall rearing in the hot-plate test were recorded in O3 prenatally exposed mice. Except for the first open-field test, altered responses were observed only in animals exposed at the intermediate concentration of the gas. Adaptation and/or onset of compensatory mechanisms might be responsible for the lack of linear dose-response relationships.