Automated analysis of DNA damage in the high-throughput version of the comet assay.

Recently a high-throughput version of the comet assay was developed using a special 96-well multichamber plate (MCP) [1]. In this version, the electrophoresis is performed directly on the MCP, which makes transferring of cells to microscope slides unnecessary. In order to facilitate the scoring procedure we adapted an automated slide-scanning system (Metafer MetaCyte with CometScan) to enable unattended analysis of comets on the MCP. The results of the system were compared with the data obtained with two interactive comet-assay analysis systems. For induction of DNA damage in human fibroblasts methylmethane sulfonate (MMS) or H2O2 was used. The three systems revealed similar, concentration-dependent results for all parameters tested: tail moment (tm), % DNA-in-tail and olive tail moment. Near the detection limit of 5-6% DNA-in-tail a significant difference with the untreated control was obtained by use of four parallel samples (p=0.01). With the newly developed automated analysis system, the evaluation of either 50 or 100 comets yielded similar standard errors for either treatment with MMS or H2O2, thus showing that the method is suitable to reveal the crucial low-dose effects with high precision. The results also show that the time needed for automated evaluation of comets on the MCP was reduced by a factor of 10 when compared with the time required for interactive evaluation. In summary, the high-throughput version of the comet assay combined with the automated evaluating system increased the output by a factor up to 180 compared with the standard method.

Tomographic rf spectroscopy of a trapped Fermi gas at unitarity.

We present spatially resolved radio-frequency spectroscopy of a trapped Fermi gas with resonant interactions and observe a spectral gap at low temperatures. The spatial distribution of the spectral response of the trapped gas is obtained using in situ phase-contrast imaging and 3D image reconstruction. At the lowest temperature, the homogeneous rf spectrum shows an asymmetric excitation line shape with a peak at 0.48(4)epsilonF with respect to the free atomic line, where epsilonF is the local Fermi energy.

Pairing without superfluidity: the ground state of an imbalanced Fermi mixture.

We used radio-frequency spectroscopy to study pairing in the normal and superfluid phases of a strongly interacting Fermi gas with imbalanced spin populations. At high spin imbalances, the system does not become superfluid even at zero temperature. In this normal phase, full pairing of the minority atoms was observed. Hence, mismatched Fermi surfaces do not prevent pairing but can quench the superfluid state, thus realizing a system of fermion pairs that do not condense even at the lowest temperature.

Superfluid expansion of a rotating Fermi gas.

We study the expansion of a rotating, superfluid Fermi gas. The presence and absence of vortices in the rotating gas are used to distinguish the superfluid and normal parts of the expanding cloud. We find that the superfluid pairs survive during the expansion until the density decreases below a critical value. Our observation of superfluid flow in the expanding gas at 1/kFa=0 extends the range where fermionic superfluidity has been studied to densities of 1.2x10(11) cm(-3), about an order of magnitude lower than any previous study.

Observation of phase separation in a strongly interacting imbalanced fermi gas.

We have observed phase separation between the superfluid and the normal component in a strongly interacting Fermi gas with imbalanced spin populations. The in situ distribution of the density difference between two trapped spin components is obtained using phase-contrast imaging and 3D image reconstruction. A shell structure is clearly identified where the superfluid region of equal densities is surrounded by a normal gas of unequal densities. The phase transition induces a dramatic change in the density profiles as excess fermions are expelled from the superfluid.

Vortices and superfluidity in a strongly interacting Fermi gas.

Quantum degenerate Fermi gases provide a remarkable opportunity to study strongly interacting fermions. In contrast to other Fermi systems, such as superconductors, neutron stars or the quark-gluon plasma of the early Universe, these gases have low densities and their interactions can be precisely controlled over an enormous range. Previous experiments with Fermi gases have revealed condensation of fermion pairs. Although these and other studies were consistent with predictions assuming superfluidity, proof of superfluid behaviour has been elusive. Here we report observations of vortex lattices in a strongly interacting, rotating Fermi gas that provide definitive evidence for superfluidity. The interaction and therefore the pairing strength between two 6Li fermions near a Feshbach resonance can be controlled by an external magnetic field. This allows us to explore the crossover from a Bose-Einstein condensate of molecules to a Bardeen-Cooper-Schrieffer superfluid of loosely bound pairs. The crossover is associated with a new form of superfluidity that may provide insights into high-transition-temperature superconductors.

Formation dynamics of a fermion pair condensate.

The dynamics of pair condensate formation in a strongly interacting Fermi gas close to a Feshbach resonance was studied. We employed a phase-shift method in which the delayed response of the many-body system to a modulation of the interaction strength was recorded. The observable was the fraction of condensed molecules in the cloud after a rapid magnetic field ramp across the Feshbach resonance. The measured response time was slow compared to the rapid ramp, which provides final proof that the molecular condensates reflect the presence of fermion pair condensates before the ramp.

Observation of Feshbach resonances between two different atomic species.

We have observed three Feshbach resonances in collisions between 6Li and 23Na atoms. The resonances were identified as narrow loss features when the magnetic field was varied. The molecular states causing these resonances have been identified, and additional 6Li-23Na resonances are predicted. These resonances will allow the study of degenerate Bose-Fermi mixtures with adjustable interactions and could be used to generate ultracold heteronuclear molecules.

New developments in automated cytogenetic imaging: unattended scoring of dicentric chromosomes, micronuclei, single cell gel electrophoresis, and fluorescence signals.

The quantification of DNA damage, both in vivo and in vitro, can be very time consuming, since large amounts of samples need to be scored. Additional uncertainties may arise due to the lack of documentation or by scoring biases. Image analysis automation is a possible strategy to cope with these difficulties and to generate a new quality of reproducibility. In this communication we collected some recent results obtained with the automated scanning platform Metafer, covering applications that are being used in radiation research, biological dosimetry, DNA repair research and environmental mutagenesis studies. We can show that the automated scoring for dicentric chromosomes, for micronuclei, and for Comet assay cells produce reliable and reproducible results, which prove the usability of automated scanning in the above mentioned research fields.

Condensation of pairs of fermionic atoms near a Feshbach resonance.

We have observed Bose-Einstein condensation of pairs of fermionic atoms in an ultracold 6Li gas at magnetic fields above a Feshbach resonance, where no stable 6Li2 molecules would exist in vacuum. We accurately determined the position of the resonance to be 822+/-3 G. Molecular Bose-Einstein condensates were detected after a fast magnetic field ramp, which transferred pairs of atoms at close distances into bound molecules. Condensate fractions as high as 80% were obtained. The large condensate fractions are interpreted in terms of preexisting molecules which are quasistable even above the two-body Feshbach resonance due to the presence of the degenerate Fermi gas.

Fiftyfold improvement in the number of quantum degenerate fermionic atoms.

We have produced a quantum degenerate 6Li Fermi gas with up to 7 x 10(7) atoms, an improvement by a factor of 50 over all previous experiments with degenerate Fermi gases. This was achieved by sympathetic cooling with bosonic 23Na in the F=2, upper hyperfine ground state. We have also achieved Bose-Einstein condensation of F=2 sodium atoms by direct evaporation.

Radio-frequency spectroscopy of ultracold fermions.

Radio-frequency techniques were used to study ultracold fermions. We observed the absence of mean-field "clock" shifts, the dominant source of systematic error in current atomic clocks based on bosonic atoms. This absence is a direct consequence of fermionic antisymmetry. Resonance shifts proportional to interaction strengths were observed in a three-level system. However, in the strongly interacting regime, these shifts became very small, reflecting the quantum unitarity limit and many-body effects. This insight into an interacting Fermi gas is relevant for the quest to observe superfluidity in this system.

Observation of Bose-Einstein condensation of molecules.

We have observed Bose-Einstein condensation of molecules. When a spin mixture of fermionic 6Li atoms was evaporatively cooled in an optical dipole trap near a Feshbach resonance, the atomic gas was converted into 6Li2 molecules. Below 600 nK, a Bose-Einstein condensate of up to 900 000 molecules was identified by the sudden onset of a bimodal density distribution. This condensate realizes the limit of tightly bound fermion pairs in the crossover between BCS superfluidity and Bose-Einstein condensation.

Decay of an ultracold fermionic lithium gas near a Feshbach resonance.

We studied the magnetic field dependence of the inelastic decay of an ultracold, optically trapped fermionic 6Li gas of different spin compositions. The spin mixture of the two lowest hyperfine states showed two decay resonances at 550 and 680 G, consistent with the predicted Feshbach resonances for elastic s-wave collisions. The observed lifetimes of several hundred ms are much longer than the expected time for Cooper pair formation and the phase transition to superfluidity in the vicinity of the Feshbach resonance.

Correlation of chromosomal aberrations and sister chromatid exchanges in individual CHO cells pre-labelled with BrdU and treated with DNaseI or X-rays.

PURPOSE: To analyse the correlation between chromosomal aberrations and sister-chromatid exchanges (SCE) in cells treated in G1 phase with X-rays or DNaseI. MATERIALS AND METHODS: Chinese hamster ovary (CHO) cells were labelled with 5'-bromodeoxyuridine (BrdU) for one round of replication and irradiated in G1 phase with 1.2, 2.4, 3.6 and 4.8 Gy X-rays or treated by electroporation with 1000, 2000, 3000 and 4000 units DNaseI per 800 microl electroporation buffer. Using a computer-aided metaphase relocation system first post-treatment mitoses were analysed for SCE and chromosomal aberrations allowing a precise investigation of correlation between both phenomena. RESULTS: A better correlation between aberrations and SCE was observed for DNaseI than for X-rays. X-rays induced more SCE than expected on the basis of aberrations, whereas the frequencies of SCE induced by DNaseI can mainly be accounted for by chromosomal aberrations. DISCUSSION: The results obtained with X-rays support an earlier observation that radiation induces both "true" SCE that are not related to chromosomal aberrations and result from radiation damage to BrdU, and "false" SCE that result from exchange-type aberrations. The high frequency of aberrations observed in cells treated with DNaseI and the good correlation between aberrations and SCE suggests that the endonuclease induces mainly "false" SCE.

The role of the pharmacist in humanitarian aid in Bosnia-Herzegovina: the experience of Pharmaciens Sans Frontières.

BACKGROUND: Founded in 1985, Pharmaciens Sans Frontières (PSF) is a nongovemmental organization of pharmacists involved in humanitarian aid. PSF relied on approximately 100 expatriates in 1998, which included 50 pharmacists distributed throughout 24 missions (i.e., 14 emergency, 7 development, 3 assessment). It is necessary to add 200-250 local staff to this group. OBJECTIVE: To describe PSF's mission in Bosnia-Herzegovina from 1992 to 1999 and to define the pharmacist's impact in the supply of medicines and the development of pharmaceutical care and services. RESULTS: In April 1992, at the beginning of Sarajevo's siege, PSF sent a small team of three volunteer pharmacists to Bosnia-Herzegovina. The objective of the emergency phase (1992-1995) was to set up a massive supply program of essential medicines and medical and biologic materials and to implement a distribution system based on existing health centers. The signing of the Dayton peace agreement and a progressive return to peace and stability marked the beginning of the postemergency phase (1995-1997). This phase pursued previous objectives of establishing a distribution network and added the implementation of logistic centers. PSF widened its involvement to medical laboratory analysis, production of medicines, disposal of expired medications sent during the conflict, and the implementation of a national center for quality control. Currently, the development phase's (1998-1999) objective is to provide adequate support for the reorganization of pharmaceutical care and services by establishing pharmacy work groups and developing and maintaining good relationships with the international community and Bosnia-Herzegovina pharmacists. CONCLUSIONS: Humanitarian aid is essential in major conflicts, as seen in the case of Bosnia-Herzegovina. Although it is difficult to evaluate the impact of the distribution network implemented by PSF, it allowed for a better provisioning of medications to the general population. PSF played an important role in such cases. In fact, PSF provides its pharmaceutical expertise to these embattled areas not only by offering financial support to the logistics or supplying of medications, but by offering professional support to the organization/reorganization of the pharmaceutical practice.

Influence of sodium butyrate on the induction of radiation-induced chromosomal aberrations and sister chromatid exchanges in Chinese hamster ovary (CHO) cells.

CHO cells were pre-treated with sodium butyrate (SB) for 24 h and then X-irradiated in G1. Metaphases were scored for the induction of chromosomal aberrations and sister chromatid exchanges (SCEs). The data were compared with those obtained after irradiation of cells not pre-treated with SB and showed that SB has different effects on the endpoints examined. The frequencies of dicentric chromosomes were elevated and of small acentric rings (double minutes, DMs) reduced. These results are discussed to be a consequence of conformational changes in hyperacetylated chromatin which could lead to more interchromosomal and to less intrachromosomal exchanges. SB itself induces a few SCEs but suppresses the induction of SCEs by X-rays. We assume that a minor part of radiation induced SCEs are 'false' resulting from structural chromosomal aberrations, such as inversions, induced in G1. Inversions are the symmetrical counterparts of DMs. If inversions are suppressed by SB treatment to a similar extent as DMs a small reduction of SCEs by SB can be expected.

Thiopronin reduces the frequencies of neocarzinostatin-induced chromosomal aberrations and sister chromatid exchanges in Chinese hamster ovary cells.

Chinese hamster ovary (CHO) cells were treated in the G1 phase of the cell cycle with different concentrations of neocarzinostatin (NCS) alone or in combination with N-(2-mercaptopropionyl)-glycine (thiopronin; TP). TP reduces the frequencies of NCS-induced chromosomal aberrations (CA) and of sister chromatid exchanges (SCE) significantly when added to the cultures simultaneously (TPsim), 1 min (TP1) or 10 min (TP10) after the addition of NCS. The addition of TP 30 min (TP30) or 60 min (TP60) after NCS reduces the frequencies of SCE, but not of CA. Our results indicate that the induction of CA and SCE by NCS is partially based on different mechanisms.

Computerized image analysis of sister chromatid exchanges in Chinese hamster ovary cells.

In this paper we show that sister chromatid exchanges (SCE) can be analysed by computer-supported determination of areas and shapes of darkly stained chromatid sections. In combination with appropriate statistical analyses, our method leads to measurements of SCE frequencies in neocarzinostatin-treated Chinese hamster ovary cells which are nearly as precise as the ones obtained by classical microscopical analyses. We discuss our data as an approach to the development of a fully automated SCE scoring system.

Neocarzinostatin induces chromosomal aberrations and sister chromatid exchanges in Chinese hamster ovary cells.

Chinese hamster ovary (CHO) cells were treated with neocarzinostatin (NCS) and analyzed for chromosomal aberrations and sister chromatid exchanges (SCE). After treatment the cells were recovered for 9, 20, 26 or 30 h. NCS induces chromosomal aberrations and SCE. SCE were much more frequent in cells with chromosome type aberrations at 20 h recovery time than in those with chromatid type aberrations at 9 h recovery time. In second post-treatment cells at 26 or 30 h recovery time NCS induced chromosomal aberrations but only few SCE.