DELTA-HEART: Physiological parameters that predict orthostatic intolerance after spaceflight.

PI: J.M. Karemaker (NL), Co-I’s: J. Gisolf (NL), W.J. Stok (NL), J.J. van Lieshout (NL), C. Gharib (Fra), Ph. Arbeille (Fra), J.Ph. Saul (USA).

The objective of the HEART experiment was to predict orthostatic intolerance, i.e. the inability to stand upright, of astronauts who have stayed for some time in a weightless environment. The hypothesis of this experiment was that such predictions could be made from a series of physiological measurements, which are done in a preflight tilt protocol.
Test outcome was the duration of the post-flight stand test, where astronauts, immediately after return from Space and immediately after waking up in the first few days post flight, are asked to stand relaxed for a maximum of 10 minutes. Dizziness, fainting or the inability to finish the stand test was considered definite proof of orthostatic intolerance.

The Dutch cosmonaut on the Delta flight was the last test person to be included in the HEART-protocol. Earlier, 7 astronauts of the Columbia flight, 3 cosmonauts of the Odissea mission and 1 of the Cervantes mission had been included, making a grand total of 12 test subjects. Due to the Columbia disaster, only 5 have made it to the crucial post flight stand tests.

Of these 5 test subjects none fainted during our observations, or was unable to finish the stand tests. On average 40% of returning astronauts are reported to suffer orthostatic intolerance in varying degrees of severity. Statistically 0 out of 5 in the present protocol does not contradict the general trend (p=0.078), but it made us look more carefully at the blood pressure and heart rate recordings during the stand tests, to look for early signs of orthostatic intolerance, rather than full blown syncope.

Figure 1 shows representative portions of the continuous recording of blood pressure (ABP) and heart rate (HR) at the landing site, supine and upright. It is obvious from the supine recording that the subject is doing fine, but for obvious reasons he is not in a very relaxed state (high and variable HR, variable ABP). However, on standing up HR goes to a very high value and blood pressure shows signs of low cardiac output, presumably due to insufficient cardiac filling, and a very large variability, considered signs of impending syncope (which did not occur).

HEART- Figure 1.

Preliminary conclusions:
All postflight recordings in 5 returning cosmonauts show, in varying degrees, signs of orthostatic problems in the early postflight period. These signs ameliorate, but do not disappear in the first week postflight. The usual post flight stand test (orthostatic intolerance: yes/no) is too crude and should be refined to take blood pressure lability and other signs of low cardiac output into account. Any computer model of the circulation should be able to show realistic values of heart rate and blood pressure variability at low cardiac filling.

DELTA-CIRCA: 24-Hour pattern of blood pressure and heart rate in weightlessness

PI’s: J.M. Karemaker (NL), C. Gharib (Fra); Co-I’s: J. Gisolf (NL), W.J. Stok (NL), G.A. van Montfrans (NL), M.A. Custaud (Fra), A. Aubert (B).

The CIRCA experiment aimed to measure the pattern of blood pressure and heart rate in an astronaut during a 24-hr period using the combined data from measurements of blood pressure at the upper arm (discontinuous recording, once every 15 minutes, but continuing for 24 hrs) and at the finger (continuous recording, but only at selected periods of the day/night period). On the Delta flight this experiment would elaborate on data obtained in 1 earlier cosmonaut on the Cervantes mission (upper arm measurements only) and on 7 test subjects undergoing a 42 days Space flight simulation (Head Down Tilt-1994, Toulouse, weekly continuous finger pressure recordings for 24 hrs).

The experiments aimed to settle a number of questions related to the condition of the cardiovascular system under microgravity conditions:
1. Is the day/night difference in blood pressure and heart rate in Space the same or less compared to Earth-bound conditions?
2. What is the physiological basis of the day/night difference on Earth and in Space, i.e. is it due to cardiac or vascular factors or both?
3. What is the condition of the subject’s autonomic nervous system in Space, compared to Earthly circumstances?

The first two questions should follow from the analysis of the upper arm cuff measurements, combined with the continuous finger blood pressure, the third one by a series of function tests that the subject was to perform on two moments of the day.

Poor functioning of the upper arm blood pressure device has hampered the experiments. Not only was the cuff inflation more painful than normal for this type of device, moreover was the device difficult to handle and very sensitive to technical failure. This has caused an undue amount of data loss in the two Soyuz missions where it has been used. Pain during cuff inflation, moreover, has probably caused more disturbances of the sleep period than usual, and has probably made the absolute differences between day and night less than during a normal, healthy sleep.

The change in autonomic condition from Earth to Space, as measured in the Delta mission, adds to a series of subjects on earlier flights. The French and Belgian group members will deliver these data.

The day/night difference in blood pressure virtually disappeared in Space, while the drop in heart rate remained, confirming our earlier observations in HDT-94. More detailed analysis of the finger blood pressure signal elucidating the physiological mechanisms behind these changes will be presented at the December 2004 Delta meeting.

Delta-MOP: Investigation of vestibular adaptation to changing gravity levels on earth.

Suzanne Nooij (email: nooij@ tm.tno.nl), Jelte E. Bos (PI), Eric Groen, Willem Bles. TNO Human Factors, Soesterberg, The Netherlands

Background: During the first days in space 50-80 % of the astro- and cosmonauts suffer from the Space Adaptation Syndrome (SAS). The symptoms of SAS, like nausea and dizziness, are especially provoked by head movements. Although it is generally agreed that the vestibular system is involved in causing SAS, no distinct clue has been found to its aetiology , the individual’s susceptibility, and its predictability. Susceptibility to SAS does not correlate with susceptibility for motion sickness on earth. However, astronauts have mentioned close similarities between the symptoms of SAS and the symptoms they experienced after a 3G centrifuge run on earth (Sickness Induced by Centrifugation, SIC). This suggests that a gravity transition from 3 to 1 G provokes the same effects as a transition from 1 to 0 G, implicating a general vestibular adaptation mechanism to changing G-levels.
Objectives: This study aims to further the insight in the process of vestibular adaptation to G-transitions. Two important parameters are the perception of body motion and attitude during the adaptation process. A second objective is to investigate the correlation between susceptibility to SAS and to SIC.
Methods: During several space missions the correlation between susceptibility to SIC and SAS has been investigated (1). Since head movements are shown to be provocative, this provocativeness was taken as an indicator for SIC and SAS susceptibility. Susceptibility to SIC was assessed after a 1 h centrifuge run at 3Gx, susceptibility to SAS during the mission. Within the framework of the 2004 Delta Mission, vestibular adaptation was addressed for 2 astronauts in four vestibular function tests carried out about a 1h 3Gx centrifuge run (-1h, +0h, +2h, +4h). The tests included motion perception and sickness ratings, stabilometry in a tilting room, subjective vertical measurements in a tilting chair and eye movement registrations (orientation of Listing’s Plane).
Results: At present, a total of 10 astronauts were tested both in the centrifuge and in space. We found a positive correlation between susceptibility to SIC and SAS: 4 astronauts were both susceptible to SIC and to SAS, 6 were not. The vestibular function tests showed that postural stability was decreased after the centrifuge run in one SIC-susceptible subject and unaffected in the other non-susceptible subject. No clear effect of the centrifuge run on tilt perception could be established. Although the data on the orientation of Listing’s Plane is still preliminary, it suggests an effect of the centrifuge run on the elevation angle of the Listings Plane.
Conclusions: The positive correlation between susceptibility to SIC and to SAS is in agreement with the hypothesis that SIC and SAS share the same underlying mechanism. This makes long duration centrifugation a valuable tool for investigating vestibular adaptation to G-transitions on earth. The gained knowledge can be implemented in a general model of vestibular adaptation. The vestibular tests showed that several vestibular driven processes are affected by the gravity transition. However, further testing is needed to identify key adaptation parameters.
1. Bles,W. de Graaf,B. Bos,J.E. (1997) A sustained hypergravity load as a tool to simulate space sickness. J.Gravit.Physiol. 4:1-4

DELTA-OLP :The collinearity of Listing’s plane and the vestibulo-oculomotor response in microgravity.

A.H. Clarke¹, T. Haslwanter², J.E. Bos³ 1: Vestibular Research Lab, Charité Medical School, Berlin, 2: Dept. of Neurology, Univ. Zurich, CH, 3: TNO-Human Factorsd, Soesterberg, NL.

Spatial orientation and movement requires that our sensory and motor systems, and their representation in the central nervous system, are correctly matched. Throughout evolution on Earth, the gravity vector has provided an important reference for the alignment of these various systems.
In particular, the otolith organs in the vestibular system are responsible for signalling the orientation of the head to the direction of gravity. This has been confirmed by numerous experiments that have demonstrated a “re-calibration” of the vestibular system in microgravity. The aim of the proposed experiment is to determine to what extent the oculomotor system and the vestibular system – which are normally collinear - are modified in prolonged microgravity. The frame of reference for eye movement control is defined by the so-called Listing’s Plane and that of the vestibular system by the orientation of the 3D- vestibulo-ocular reflex (VOR).
The working hypothesis is that in microgravity, due to the loss of the gravity vector as reference, the collinearity of Listing’s Plane and the VOR co-ordinate frame will be reduced.
Listing’s Plane is estimated by precise measurement of eye position while the subject fixates various points in the visual field. The 3D-VOR co-ordinate frame can be determined by analysing eye and head movement during active head movements around the yaw, pitch and roll axes. The DLR Eye Tracking Device (ETD), used for the first time on the Delta Mission, provides for accurate three-dimensional measurement of eye and head position.
Four pre-flight BDC sessions, three inflight sessions and 7 postflight sessions were performed successfully. To date, the recorded sequences for the evaluation of Listng’s Plane have been analysed. The evaluation of the VOR sequences is in progress.
The pre-flight BDC trials demonstrated that the orientation of was tilted back by 3 to 5°. Contrary to expectation, it was found that Listing’s Plane was tilted forward during the inflight period in microgravity. During postflight measurements the orientation returned to the pre-flight values. This finding is novel, and suggests that not only the vestibular, but also the oculomotor system depends on the gravity vector as reference.

DELTA-SAMPLE: Molecular and physiological analysis of bacterial samples isolated from manned space craft.

Hermie J. M. Harmsen¹, Erwin C. Raangs¹, Gjalt W. Welling¹, Paolo Landini², Luc van den Bergh³, Janneke Krooneman⁴. ¹University of Groningen, ³Dutch Space, ⁴Bioclear, The Netherlands; ²University of Milan, Italy.

The SAMPLE experiment that was performed during the Delta mission was succesful.
The objectives of the experiment were: (i) evaluate which microbial species might benefit from growth conditions in life support systems and (ii) investigate if adaptation occurs in the mechanism of microbial adhesion due to microgravity. For the first objective samples were taken with cotton swabs from the interior of the International Space Station (ISS) and these were analysed on earth (Fig 1). For the second objective cells of the bacterium Escherichia coli were grown in the ISS. The cells were exposed to these space-conditions for ten days and subsequently transported to earth to analyse if adherence properties of the bacterium adapted to microgravity: no adhesion adaptation was noticed after the ten days that the bacteria were exposed to microgravity.
The swab samples of the ISS were analysed with different microbial methods: by conventional culturing and by DNA analysis methods.

	Figure 1. All the SAMPLE swab-stick tubes after return to earth.
	Figure 2: A plate with culture medium for yeasts (pink colonies) and fungi (black moulds). The inoculum originated from a wall panel in the ISS.

Culturing showed that some of the sites were colonized with bacteria, yeasts and fungi. However, the actual numbers of microbes present could be higher than was found, because storage and transport may have influenced the quality of the samples that may have caused death of part of the microbes. The DNA methods are far less hampered by cell death and enabled a very accurate analysis of the samples. One of these methods, quantitative real-time PCR, showed that surfaces were colonized with human bacteria, such as staphylococci, in some samples in relatively high numbers. This was in concordance with the culture results although the culturing numbers were a few orders of magnitude lower. Another method, fluorescence in situ hybridisation, showed a similar picture of high numbers of bacteria, human bacteria, yeasts and fungi. Our results show that with these methods the microbial contamination of the ISS can be monitors and that in this way hygiene and health status of the ISS can be optimised. Both DNA methods are robust and can be automated, and have a potential to be used in space.

DELTA-MUSCLE: Study of lower back pain in cosmonauts during spaceflight.

CJ Snijders¹ and AL Pool-Goudzwaard¹, CA Richardson² and JA Hides². ¹Erasmus MC, University Medical Center Rotterdam, Dept. of Biomedical Physics and Technology, Rotterdam, NL, ²University of Queensland, Dept. of Physiotherapy, Queensland, Australia.

Experiment objective and expectations: Aim of this study was to obtain data about the development of complaints during flight on a day-to-day basis. Based on the biomechanical model it was expected that a) low back pain could develop at the site of the iliolumbar ligaments (the iliac crest) and b) a situation of combined low back pain and constipation could develop.
Method: The questionnaire started with "Did you experience pain today in the lower back?" and if the answer was "Yes", questions about type and intensity of pain were completed with the use of a visual analogue scale. We also asked if the back was painful almost all the time, what provoked the low back pain and if it was possible to relieve the pain. Final question was if change in bladder or bowel function occurred. The questionnaire was incorporated in the general mission logbook and had the following schedule: Pre-flight: At L-10 +/- 5 baseline data were collected with the questionnaire; In-flight: Completion of the questionnaire at the end of every flight day ; Post-flight: At R+10 +/- 5 data on return to gravity load were collected with the questionnaire. The Ethical Committee approval was obtained from the institutional Review Board of the University Medical Center Rotterdam. Informed consent was signed. Because the questionnaire was anonymous, the results of this single case study are presented in a global form and in consult with the cosmonaut.
Results: Pre-flight no complaints were recorded. During the first days in-flight, with a peak on day 4, back pain bilateral in the region of the iliac crest was recorded, together with constipation. Both complaints disappeared in a few days. Of special interest is, that pain relief could be realised by stretching of the back, i.e. realizing a lumbar lordosis. With the pain relief also the constipation disappears.
Conclusions: Combination of the recordings during flight with the unilateral low back pain recorded post-flight after 4 days suggests a) that the in-flight pain (modest, 3 on the VAS-scale) cannot have been caused by trauma during the start procedure and b) cannot have been discogenic.
From this study we conclude, that the answers to the questionnaire are in favour of our biomechanical model. This regards the recorded complaints as well as the countermeasures practiced by the cosmonaut. Therefore we regard this project as successful.
We are aware of the fact that the results of this single case study must be interpreted with reserve, but they form a strong base for future studies with series of cosmonauts who stay in the ISS during several months. Results of future studies can have a large impact for countermeasures in-flight and treatment of low back pain patients on earth.

Deep muscle corset being subject to wasting and loss of co-ordination in the microgravity environment. Comprehensive Biomechanical model on sacroiliac joint stability with transversely oriented abdominal muscles (transversus abdominis), back muscles (sacral part of multifidus) and pelvic floor muscles (coccygeus) (DELTA-MUSCLE).

DELTA-ARGES: A tomic densities measured Radially in metal halide lamps under micro Gravity conditions with Emission Spectroscopy.

Gerrit Kroesen a.o.¹ , Marco Haverlag a.o.² , ¹TU Eindhoven, NL, ²Philips CDL, Eindhoven, NL

The experiments in the ARGES project were a 100 % success. All foreseen measurements have been taken and all data was transported back to the experimenters. Already during the experiment operations, the results proved to be very surprising. Whereas the instabilities in the lamp were expected to be shaped as a rotating helix, they appeared to be a singly bent curve which is not rotating. Analysis afterwards has indicated that the rotation is caused by convection solely and that the curving is caused by self-generated magnetic fields. For one condition, residual gravity caused a very slow rotation.
As expected, the axial de-mixing did not occur during the Delta mission experiments, so the radial de-mixing can indeed be studied undisturbed. The analysis of the spectra is well underway.

On the ground Axial and radial de-mixing

In space

On the ground

In space t1

In space t2

In space t3

DELTA-SUIT: A Vibrotactile Orientation Support Tool.

Jan B.F. van Erp. TNO Human Factors, Soesterberg, NL & Dutch Space, Leiden, NL.

In everyday life, we gather information about the world around us via different sensory systems, including our sense of touch. In a micro?gravity environment, astronauts lack specific sensory information. Especially the vestibular and cutaneous senses are affected. For example, there is no pressure on the sole of the feet when standing, there are no G?forces to overcome when positioning limbs or that pulls clothing to the skin, and no gravitational acceleration to which the otoliths respond (in microgravity, the otoliths are effectively unloaded and cannot provide information about static head orientation). This sensory deprivation has several consequences for astronauts. Amongst other things, it affects the way astronauts perceive their subjective vertical. In the SUIT project, we developed a orientation support tool based on providing an artificial gravity vector by a localised vibration on the torso (see Figure 1, 2). Such a support tool is intended to be used in challenging situations, such as Extra Vehicular Activity, in which the astronaut’s degraded orientation awareness may be critical with respect to safety, performance or comfort. To investigate the potential of the support tool, we designed an experiment in which the astronaut performed a set of tasks under different conditions. These tasks were done under normal, non-challenging conditions and were intended as a proof-of-concept.

Figure 1 (top). The SUIT concept. A vibration is given at the location where the artificial gravity vector intersects the astronaut's torso. The direction of "down" (i.e., the artificial gravity vector) can be freely chosen by the astronaut.

Figure 2 (right). The SUIT hardware consists of a vest with 56 vibrating elements, a sensorpack with three gyroscopes, batteries and a control unit worn on the wrist.

Figure 3 (left). One of the results of the experiment in which the availability of the support tool results in performance increase, in this case the time the astronaut needed to orient himself.

The results show that the support tool results in: (a) a faster completion of the tasks, (b) better task performance, and (c) tasks being subjectively rated as easier than the control conditions in which the support tool was off. An example is given in Figure 3 in which the Reaction Time to indicate where down is after being brought into an unknown orientation is significantly reduced in the conditions with SUIT support.

We concluded that the support tool is able to enlarge the astronaut’s orientation awareness and that the experiment was both a successful proof-of-concept as well as a successful technology demonstration. A further identification of the challenging and critical situation in which the tool may have a surplus value is required.

DELTA-LSO: Lightning and Sprite Observations.

Elisabeth Blanc¹, Peter Van Velthoven². ¹CEA-France, ²KNMI – Netherlands.

The experiment LSO is dedicated to the optical study of sprites from the International Space Station.
Sprites discovered in 1989, have the appearance of a luminous glow extending from 30 to 90km altitude above thunderstorms. Their duration varies from a few milliseconds up to a few hundred milliseconds. They occur with other emissions called jets, elves and halos, in relation with intense electric fields between the thunderclouds and the ionosphere (above 100 km). This direct coupling between the active storm cells, the thermosphere and the ionosphere and the considerable energies involved gives rise to processes that have been unsuspected until now regarding the space plasmas as well as the chemistry and dynamics of the middle atmosphere. These processes can have a significant effect on the Earth’s magnetosphere, in particular by modifying the source terms and loss of the radiation belts.
The objectives of the LSO experiment are to validate a new measurement concept for future measurements of sprites from space at the nadir based on a spectral differentiation between sprites and lightning flashes. This measurement concept will be used by the microsatellite Taranis dedicated to the study of sprites and associated emissions. The experiment is also dedicated to expanding global statistics about the spatial and temporal distributions of the sprites.
Observations are performed by two micro-cameras, one in the visible and near infra red, the other equipped with a moderately wide band filter at 761 nm. This filter includes the most intense N2 1P emission of the sprites and partly the oxygen absorption A band of the atmosphere. The light emissions from sprites occurring in the middle and upper atmosphere are then differentiated from the emissions from lightning, occurring more deeply in the atmosphere and then more absorbed.
The first experiments were performed in the frame of the mission Andromede. Experiments continued in the frame of a collaboration program with Russia and of the ESA missions Odissea, Cervantes and Delta. However, these last data are unfortunately not exploitable for sprite studies because of a problem with the camera equipped with the filter. A summary of the observations of sprites by LSO at the nadir obtained up to now will be presented with the first statistics about the respective intensities of lightning and sprites emissions and sprite global occurrence.

Example of sprite observation at the nadir by LSO.

DELTA-MOT: DSM Mouse Telemetry System.

Gerard van Essen, Mans Jansen, Telemetronics, Heteren, NL.

Goal of the experiment:
- To test the accelerometers of STAR under microgravity conditions with the same electrical circuit functions
- To get the electronic circuit Space qualified.
- Test is performed by predefined arm movements during a short time for the three orthogonal axis.

Crew In flight operation.

The right graph shows the experiment’s results performed on earth, the left graph the results performed in the ISS under micro gravity. Sequentially, the MOT is moved as indicated in three orthogonal positions (to test the three orthogonal accelerometers). From the graphs it becomes clear that the accelerometers act the same way on earth as under micro gravity. Unfortunately, the X-axis accelerometer did not respond during the space experiment. The only logical conclusion is that a malfunction occurred in the X-axis accelerometer or in the subsequent electronics. Further, the vertical lines at the left side of the space graph do not occur on the terrestrial graph. They may be caused by repeatedly on and off switching before the tests.
Conclusion: The piezo electric accelerometers are suitable for use in the STAR mouse implant under development now.

DELTA-HEAT: Zero G characterisation of Aluminium grooved Heat Pipe.

L. Barremaeker¹, G. Grommers², LC Legros³. ¹Euro Heat Pipes SA, ²Dutch Space, ³ULB-MRC

The “HEAT experiment” was part of the European Space Agency - DELTA mission (“Dutch Soyuz Mission”) to the International Space Station. The experiment has been developed in the frame of the pre-development activities for the @Bus new high power telecommunication platform.
The experiment has been launched with a Russian Progress launcher on the 29 January 2004. The first session of the HEAT runs was performed by the ESA Dutch astronaut
Mr André Kuipers (April 2004) while the second session was performed during Increment 9 by the US astronaut Mr Mike Fincke (Sept. 2004) during the so called “Saturday science” activities.

Main scientific objectives: · Full characterization of the heat transport performances of a standard EHP aluminium re-entrant grooved heat pipe type AG110 (filled with NH3) under microgravity conditions, by deriving the maximum sustainable heat flux (heat pipe burn out limit) and heat transfer coefficient for six specific operational modes. · Validation of the existing mathematical hydraulic model that is used to evaluate the performances of a new generation of high performance heat pipes.
Results: Typical burn-out conditions were analyzed in order to derive the maximum heat transport capabilities of the HP. The heat transfer coefficients were also derived before the burn-out conditions.

Results summary : · For the heat transport capability, it can be noticed that the performances of the AG110 in 0G are equivalent or higher than the 1G horizontal conditions. · For the heat transfer coefficient, a very significant improvement is observed in microgravity. For equivalent test configurations, the improvement factor is from 2 to 2.3 times better in microgravity. · The preliminary software correlation shows that the EHP 0G predictions are in line with the measurements (about 14%).
Conclusions : Very promising results were recorded and shows good correlation with the heat pipe involved physical laws and predictions. Further tests in microgravity are now needed to cover HP with larger diameter and full operational temperature range.

DELTA-ICE-1st : International Ceonorhabditis elegans Experiment First flight

Science Goals and Objectives

The first International C. elegans Experiment (ICE-first), investigates several scientific domains. This unique opportunity provided to the science community gathered by the French space agency (CNES) and kindly offered by the European Space Agency and the Space research organisation of the Netherlands (SRON) is welcoming groups of scientists from France, Canada, Japan and United States of America.
Caenorhabditis elegans is a nematode (Phylum Nematoda) measuring around 1mm and living naturally in soil. It is of no economic importance to man. It has reproductive, nervous, muscular, and digestive systems. It is a very simple organism made of a fixed number of somatic cells (959). Its life span is about 2-3 weeks and in the liquid medium at 25°, the life cycle is around 5 days. The genome is totally sequenced (100 000 000 of base pairs and around 20 000 genes). Numerous mutants are available. It is used as a model system for various medical pathologies and was the subject of the 2002 Nobel Prize in Medicine or Physiology because the process of apoptosis or programmed cell death was discovered while studying C. elegans development. Around the world many hundreds of scientists are working full time investigating its biology. C. elegans is about as primitive an organism that exists which nonetheless shares many of the essential biological characteristics that are central problems of human biology. The worm is conceived as a single cell which undergoes a complex process of development. Then lineage of the 959 cells is fixed and well known allowing a huge number of investigations in fundamental biology. Scientists study embryogenesis, morphogenesis, development, nerve function, behavior and aging. C. elegans may be handled as a micro-organism. Thus it provides the researcher with the ideal compromise between complexity and tractability. Different studies have been performed during the DELTA mission.

1. Studies on genome stability (Ann Rose, Canada).
We will take advantage of the suitability of genetic approaches on C. elegans to address the genome stability question by analysing the distribution of an antigen (mdf-2) which is essential for the normal divisions of the cells. An alteration of the production of this antigen will lead to the accumulation of defects including chromosome abnormalities, X-chromosome non-disjunction or loss, problems in gonad development, and embryonic lethality (Kitagawa & Rose, 1999). Another way to evaluate the effect of radiation is to study the G-tracks which are present in the genome.
The preliminary results have been presented at the ASGSB meeting in 2004 (Worms in Space? A Model Biological Dosimeter; Yang Zhao, Robert Johnsen, David Baillie and Ann Rose).

2. Studies on muscle growth and survival (Catharine Conley USA; and Laurent Ségalat, France).
Microgravity has an important impact on muscle physiology and growth. C. elegans has muscles which are analogous to vertebrates in terms of composition and basic organisation. Investigations on C. elegans would benefit from a large number of mutants available in this organism. In this first study, emphasis would be put on two sets of genes : the localization of Tropomodulin proteins and other contractile proteins of muscle have been investigated. The phenotype of animal mutants for genes encoding protein involved in muscle survival has been analyzed,specifically the dystrophin, the product of the gene mutated in Duchêne Muscular Dystrophy. The effect of microgravity on other mutations affecting C. elegans muscle survival (including MyoD, perlecan, titin) are under study using immucytochemistry and microscopy..

3. Whole-genome microarray analysis of responses to spaceflight in C. elegans (Catharine Conley, Stuart Kim USA)
The proposed experiment involves performing RNA expression analysis using a microarray designed to probe for nearly every gene in the genome. Although microarray analysis does not provide detailed information concerning the function of any particular gene, its acts as an efficient indicator of which genes might be interesting to study further, because they show altered expression in response to the treatment. Two hypotheses that can be tested directly using microarray data a) that radiation-repair genes will be up-regulated, and b) that genes involved in muscle specification and contractility will be down-regulated. Additional hypotheses concerning worm 'immune' function and aging can also be tested, by determining the expression of genes known to be involved in those physiological processes. The sample received from space and ground experiment are under study at ARC.

4. Morphometry of larval C. elegans development during spaceflight. (Catherine Conley, Beverly Girten, USA) In liquid CeMM, the media used in the ICE flight, larvae shed cuticles as they molt and progress to the next developmental stage are measured to study the range of lengths exhibited by shed cuticles in media from cultures that have been returned alive. The distribution of length data will indicate the number and progression of larval moults during development in space.

5. Effect of space flight on cell migration and muscle cell in C. elegans development. (Hiroaki Kagawa, Noriaki Ishioka, Japan) C. elegans has two muscle tissues; pharynx for feeding and body wall muscle for locomotion. The both correspond to heart and skeletal muscle of vertebrates. Recently we found that muscle filament gene defect affect not only muscle function but also muscle development. Additionally these mutant animals have abnormal distal tip cell migration during the worm development. Abnormal cell migration can easily be seen under microscope. For this experiment, we use wild-type and thick filament abnormal mutant (unc-15), which produce muscle filament but decreased function and have abnormal morphology of distal tip cells. Thesamples are currently under study in japan

6. Studies on germ line development including meiotic chromosomal dynamics and germ cell apoptosis under microgravity condition (Atsushi Higashitani, Noriaki Ishioka, Japan) In C. elegans, the sequence of changes in chromosomal morphology during meiotic prophase 1, the oocyte maturation and the germ cell apoptosis can be observed, and the molecular mechanisms underlying these phenomena can be investigated with genetic approaches. We will analyze the effects of microgravity on these phenomena using the N2 wild-type and ced (cell death) mutant (ced-1) strain. This experiment will require 100 to 1000 animals in each strain at mixed developmental stages, in addition to 1G control of each sample in space) fixed in flight for microscopic observations staining with DAPI and several antibodies (histone H3 phosphorylation and methylation, activated MAPK etc…).

7. Analysis of the aging related protein aggregation and sarcomere integrity (Shuji & Yuko Honda, Noriaki Ishioka Japan) To examine the effects of space on protein-folding homeostasis in muscle cells, we will analyze the aggregation of polyglutamine (polyQ) in body wall muscle cells, using transgenic C. elegans (N2; Punc-54)expressing polyQ-YFP (yellow fluorescent protein) and also daf-2(e1370) lifespan-extension mutant. We will also analyze sarcomere orientation in the muscle of transgenic C. elegans (N2; Punc-54) expressing GFP (green fluorescent protein) in body wall muscle cells.

8. Description of the Experiment
To fly the animals have adapted to the liquid medium specially designed and provided by NASA. Six days before the launch the strains have been prepared for the flight by the investigators in the facility of the GSBMS (Groupement Scientifique pour la Biologie et la Médecine Spatiale) in Toulouse. According with a define scenario, some of the samples have been placed in culture chambers (CCA) which enable a fixation by the end of the flight. The others have been packed in 5.0 and 2.5 ml culture bags.
They have been all togather then hand carried to the launch pad in Baikonour under controlled temperature (12°C). After a final check of the samples in culture bags by a scientist, they have been 12 hours before the launch in vented Experiment Containers kindly provided by Prof. E. Horn from Ulm (Germany).
The samples have been delivered to the mission management to be placed in Kubik Topaz with the other experiments and the temperature was set at 20°C. The launch was uneventful for the worms. Three days after the launch, the samples have been transferred to the Kubik Amber but the centrifuge supposed to hold three small containers did not work. April the 25th the containers ICE-01 et ICE-03 have been fixed while ICE-02 and ICE-04 have been fixed April 27th.
Right upon the landing in Kazahkstan, the containers containing the culture Bags have been open and some filmed to evaluate the behaviour of the animals. The small bags containing the culture of the worms have been then either frozen or refrigerated till their return in Toulouse two days after the landing.
All the samples have then been forwarded to the investigators and are now analyzed in the laboratories. Most of the samples are suitable for the scientific purposes and the main results are planned

9. Acknowledgements
The science team acknowledges the space agencies which are allowing this flight opportunity as “l’Agence Spatiale Canadienne” (ASC), the “Centre National d’Etudes Spatiales” (CNES), the European Space Agency (ESA), the Japan Aerospace Exploration Agency (JAXA), the National Aeronautic and Space Administration (NASA), and the Space Research Organisation of the Netherlands (SRON).

DELTA-ACTIN : Role of microgravity on actin metabolism in mammalian cells.

M.J.A. Moes, J.J.M. Bijvelt and J. Boonstra. Utrecht University

Previous experiments have indicated that microgravity conditions influence cells most likely on the level of the actin microfilament system. It is the aim of this project to identify the gravity sensitive component that underlies the actin filament metabolism. The possible target of gravity will be determined by studies using the random positioning machine (RPM), centrifuge and sounding rocket conditions. A first series of experiments was performed on the Delta mission.

In preparation for the Delta mission, investigations were performed to select a model that can stand the conditions during the actual microgravity experiments. First of all a suitable container was selected. Based on criteria like reliability, biocompatibility, reproducibility, etc. a decision was made in favor of the Plunger Boxes of CCM. Mouse fibroblasts, named C3H10T1/2, were selected as cells to be used for their actin morphology and strong survival rates. These cells can be growth factor-starved and subsequently stimulated with platelet derived growth factor (PDGF). This results in spectacular changes in actin morphology. Five minutes after stimulation ruffles are induced on top of the cells. These ruffles are the result of local induced actin polymerization causing dynamic surface extensions of the membrane. Moreover, the number of stress fibers is decreasing after PDGF stimulation. Stress fibers are components of the cytoskeleton and extent from special junctions in the cell membrane, called focal contacts, to the cell nucleus or to other focal contacts. These stress fibers form in response to tension generated across a cell. The whole process is transient and can be followed in time. During the Delta mission the cells were stimulated for 0,5,10 and 30 minutes. The cells were growth factor-starved for up to 6 days, survived in the containers and were stimulated in the containers. A similar set of experiments was performed at 1G as a control.

Also simulated microgravity is used in our studies, by using a random position machine. For these experiments a device, named COBRA, was developed by Dutch Space and supported by DESC to be able to perform experiments while rotating the RPM.

Cells were stained with Phalloidin, labeling polymerized actin (F-actin).
1. Growth factor-starved cell, showing abundant stress fibers.
2. PDGF stimulated cell after 5 min., showing circular ruffling.
3. PDGF stimulated cells after 30 min., showing remnants of ruffles

Unfortunately, there were no results obtained in microgravity due to a combination of technical problems and unfulfilled temperature requirements.

DELTA-FLOW: Microgravity and osteocyte mechanosensitivity.

Bacabac RG¹, van Loon JJWA^1,2, de Blieck-Hogervorst JMA¹, Semeins CM¹, Tan D¹, Vatsa A¹, van Duin MA¹, Klein-Nulend J¹. ¹Department of Oral Cell Biology, ACTA-Vrije Universiteit, Amsterdam, NL, ²Dutch Experiment Support Center (DESC), Vrije Universiteit, Amsterdam, NL.

The main scientific objective of the FLOW experiment is to test whether the production of early signaling molecules that are involved in the mechanical loading-induced osteogenic response (nitric oxide and prostaglandin E2) by bone cells is changed under microgravity conditions compared to 1xg conditions.
Osteocytes, osteoblasts, and periosteal fibroblasts, were isolated from chicken skull and incubated in plunger boxes (developed by Center for Concepts in Mechatronics (CCM), Nuenen, NL) using plunger activation events for single pulse fluid shear stress stimulations. Cultures in-flight were subjected to microgravity and simulated 1xg level by centrifugation. Ground controls were subjected to identical culture environment and fluid shear stress stimulations.
Due to unforeseen hardware complications, results from in-flight cultures are considered lost. Ground control experiments showed an accumulative increase of nitric oxide in medium for osteocytes (as well as for osteoblasts and periostial fibroblasts, figure 1). Data from the online-nitric oxide sensor showed that the nitric oxide produced in medium by osteocytes increased sharply after the first and second pulse shear stress stimulations (figure 2, shows data from one culture).
In conclusion, preparations for the FLOW experiment and preliminary ground results indicate that the FLOW setup is viable for a future flight opportunity.

Figure 1. Accumulative nitrite concentration in the medium. Nitrite, the stable metabolite of nitric oxide, increased for all cell types 2½ hrs (III) after the 1st (I) and 2nd (III) pulse shear stress stimulations. Values are mean ± SEM. OCY, osteocytes; OB, osteoblasts; PF, periosteal fibroblasts.

Figure 2. Nitric oxide (NO) production by osteocytes, at the first and second pulse fluid shear stress stimulations by plunger activation (arrow) as monitored by the online-nitric oxide sensor.

Acknowledgements
This work was supported by funds from the Space Research Organization of the Netherlands (SRON) #MG-055. Special thanks to the staff of CCM, the European Space Agency (ESA), and the Russian patners (RKA-Energia).

DELTA-KAPPA : Influence of Microgravity on the Activation of NFk B.

Peppelenbosch M. University of Amsterdam, AMC, Amsterdam

The major challenge in contemporary healthcare concerns so-called auto- immune disease. Important examples of this type of disease include rheumatoid arthritis, asthma, allergies and food allergies and inflammatory bowel disease (Crohn and Colitis Ulcerosa) and all these diseases share the characteristic that the immune system overreacts to a harmless stimulus and in the process inflicts serious damage to the body. The incidence of these diseases has been rising for many decades and this increase in incidence has now reached almost epidemical proportions. Importantly, current clinical practice has only two major strategies of dealing with such disease: steroids and NSAIDs (apirin-like compounds). Unfortunately, both types of therapy have significant drawbacks and an increasing cohort of our patients does not react to these drugs. It is therefore important to devise novel strategies for interfering with the activity of the immune system. Interestingly, space flight inhibits the activity of the immune system, demonstrating that alternative roads to immunosuppression must exist. In the present project the molecular details of space-flight dependent immunosuppression are determined and these results will provide important clues to devise novel anti-inflammatory therapy for sufferers from auto-immune disease.

DELTA-TUBUL: Tobacco Bright Yellow-2 cells grown on Earth and in space.

AM Emons, B Sieberer, M Cusell. Wageningen University, Lab. of Plant Cell Biology. Wageningen, NL.

Aim of the TUBUL experiment was to study the effects of microgravity on microtubule cytoskeleton configurations in the microgravity environment on board of the International Space Station. The microtubule cytoskeleton is crucial for plant cell growth and division, and thus for plant development.
We successfully established a self-sustaining plant cell culture system for a 10 days space experiment during the DELTA mission. For TUBUL we used eight experiment units (PlungerBox Units). Each unit contained two growth compartments with plant cell suspension cultures with the cells immobilized in a thin layer of agar. The automated medium refreshment, chemical fixation, and post-fixative treatment worked perfectly well and accordingly to the TUBUL experiment time line. An important first result is that the plant cells were growing and dividing, which lead to an increase in the number of cells per culture compartment. This indicates that the microtubule cytoskeleton must have been functional, at least to some extent, during exposure of the cells to microgravity.

Figure. 1: Tobacco Bright Yellow-2 cells grown on Earth form cell files consisting of several individual cells connected to each other. The cells are almost uniformly shaped and sized, an indication that cell growth and division are not impaired.

Figure. 2 Tobacco Bright Yellow-2 cells grown for 5 days in space. To the left: The size of the cells is unusually large, the cell file in this case consists only of 2 individual cells. This could be an indication that cell division is impaired in these cells. To the right: Irregular cell shape in cells exposed to microgravity. This could be an indication that in part of the cells cell growth is affected.

A certain number of these cells showed deviations from regular growth pattern and cell size when compared with cells from ground experiments. Unfortunately, our flight samples suffered from severe post-fixative freeze-damage that occurred during sample retrieval. The insufficient quantity of well-preserved cells does not allow a detailed and reliable cell biological analysis of cells exposed to microgravity. Despite these setbacks the TUBUL experiment has a positive outcome: We established a culture system for immobilization of plant cells, which is self-sustaining for up to two weeks and which allows three different treatments per cell culture. This culture system for microgravity conditions provides openings for more extensive cell biological research in space than would be possible with whole plants.

DELTA-ARISS: Amateur Radio on the ISS

PI: G. Bertels (ARISS, Brussels, Belgium, gaston.bertels@skynet.be); ESA Contact: B. Ten Berge (ISS Education Office)

Experiment objectives and method:
The objective of this activity was to provide a live radio link from the International Space Station (ISS) to selected children in Dutch and Belgian schools; to allow them to have the experience of interacting with an astronaut in orbit around the Earth.
The selected schools were the winners of the ‘Zeg Het ISS’ competition. The competition required that children either create a picture of an astronaut on board the ISS, or write a story about an astronaut on board the ISS, depending on their age.
The 5 winning classes were invited to Space Expo to assist in the live radio contact with the ISS via radio equipment at ESA/ESTEC in Noordwijk, the Netherlands. The radio contact was provided by ARISS, Amateur Radio on the ISS, an international working group of volunteering amateur radio operators. A 10’ radio contact was established successfully during a pass of the ISS over the Netherlands on 24 April, and André answered 18 questions prepared by the children.
After a reschedule with the ground station, a second successful contact was held with the Technical University Eindhoven, Belgium on 25 April during which time André answered 20 questions.
Looking ahead:

As a result of the successful collaboration between ESA and the Amateur Radio Club during the Soyuz Missions it has been agreed that ESA will support at least two ARISS contacts per year, involving the participation of (ESA) astronauts on board the ISS.

DELTA-VIDEO-3: Educational demonstration of human physiology.

PI: S. Ijsselstein (ESA, ISS Education Office); Scientific Consultants: JJWA van Loon (DESC – Amsterdam, The Netherlands); M. Paiva (ULB – Brussels, Belgium)

Experiment objectives and methods: The main scientific objectives of this experiment were to demonstrate some of the effects of weightlessness on the human body (e.g. blood pressure and circulation, fluid shift, orientation awareness, etc.) by means of filming (with voice-over) four basic physiology demonstrations under weightless conditions on board the ISS.
For all four experiments comparable on-ground experiments have been performed by selected students in The Netherlands, France, Belgium and Denmark to familiarise students with the differences between the Earth and space environments.
In addition, a sequence of the related CIRCA and SUIT experiments were filmed to familiarise students with the actual scientific and technological hardware currently being used on board the ISS to measure blood pressure and orientation awareness respectively.

Results: All the experiments were performed and filmed successfully during the mission with the expected results, and the footage taken of both the ‘on-ground’ and ‘in space’ demonstrations has been used to develop a DVD entitled “Body Space” fitting the basic European science curriculum of the target 12-18 year old age group. Over 10,000 copies will be distributed to secondary schools across ESA Member States around February 2005 to provide teachers with a useful tool for explaining basic physiological phenomena.
For more information please visit the ESA ISS Education webiste: www.esa.int/spaceflight/education

DELTA-Bug-NRG: The effects of microgravity on the output of bacterial fuel cells.

Sebastiaan John de Vet¹& Renske Rutgers². ¹ University of Amsterdam, Human- and Natural Sciences (formerly Delft University of Technology, faculty of Aerospace Engineering), ²University of Utrecht, Utrecht School of Economics.

The BugNRG experiment was flown to the International Space Station (ISS) on flight 8S during the Dutch Soyuz Mission named “DELTA”. The experiment studies the effects of microgravity on the output of bacterial fuel cells (BFCs). A BFC is similar to an ordinary fuel cell yet utilizes a bacterial component in the anode chamber. This bacterial component converts hydrocarbons (e.g. glucose or fructose) into the components needed for the fuel cell process. Recent and past studies have shown the effects of microgravity on certain bacteria. This involvement of microgravity in various cellular and sub-cellular processes often results in changes related to reproductive rates and activity. If the bacterial component inside the BFC is indeed influenced by microgravity, a noticeable change in output and efficiency may result from this.

BugNRG utilizes the Rhodoferax Ferrireducens strain inside a two-compartment closed bacterial fuel cell. The strain converts glucose anaerobic by transferring electrons to an electron-acceptor. The anode- and cathodechamber both have a volume of 10,3 millilitres and are separated by a cation-exchange membrane. Inside the cathodechamber a solution of Potassium Hexacyanoferrate was used as electron-acceptor. In the anodechamber, a solution of glucose and growth medium was used.

Using the BugNRG experiment facility of the space-based version of the experiment, two BFCs were flown in space. The experiment qualified for a late-access requirement to reduce influences of gravity and bacterial activity as much as possible. Filling of the fuel cells was completed some seventeen hours before launch and the experiment was activated some twelve hours before launch. The facility logged the output (voltage and current) of the fuel cells and the temperature by means of an internal temperature sensor. A datalogger containing the measurements was downloaded on the Soyuz and the remaining hardware was disposed inside the Progress.

The first series of the reference experiments were carried out under similar temperature conditions as experienced onboard ISS inside a computer controlled incubator. Four fuel cells were subjected stationary to the ISS-conditions and three additional fuel cells were subjected to simulated microgravity on a Random Positioning Machine (RPM) for a duration of five days. Two back-up fuel cells prepared for flight to ISS were subjected to ambient room temperature and transportation. These fuel cells have been included into the data to partially asses the dynamic loading of the fuel cells (motion and acceleration).

An early preliminary data analyses shows the ISS as the RPM experiments to have a different behaviour with regard to the output. This behaviour differs from the normal discharge of fuel cells under the earth’s gravity as observed in the reference experiments. However, a more conclusive and extensive data-analysis is needed to fully asses the influence of microgravity on the output of bacterial fuel cells.

A second series of the reference experiments is currently being planned. The possibilities to run an experiment with BFCs subjected to hypergravity to further study the influence of gravity on bacterial fuel cells is currently being assessed.

The stars, the universe, space flight, and astronauts have always been exciting and inspiring to mankind, especially to children. Although exiting the number of pupils and students who have an interest and plan a career in the field of science and technology has been declining in many countries including the Netherlands over the years.
The educational experiment Seeds-in-Space was an effort to stimulate young people, age group 10-14 years, to participate in a science experiment during the Dutch Soyuz Mission, DELTA. The rationale of Seeds-in-Space was that it is performed on board the International Space Station, by the Dutch – ESA astronaut Andre Kuipers, and at exactly the same time, the same experiment was performed by children on ground.
We designed a rocket-shaped growth chamber in which we grew rocket lettuce (Rucola or Eruca sativa). Seeds germinated in a dark or light chamber. After 4 days the microgravity grown flight samples were compared with the 1xg cultures on ground.
Within the Netherlands a total of at least 70.000 children participated in this experiment each with a personal rocket. This is about 15% of the total population in this age group. An additional 70-80 thousand children in Germany also joined the experiment in a one-rocket per classroom setting. Also some schools in Moscow and in the Dutch Antilles participated. Seen the impact of this experiment, ‘Seeds’ may be considered for future (European) missions to ISS.

Start of the Seeds-in-Space experiment.

André checking the Seeds-in-Space rockets in flight.

In preparation of the flight we verified the growth using the Random Positioning Machine (RPM) so the actual flight results were not completely surprising. We also performed ground based tests in the Zvezda mock-up in Star City. For the larger part, the results were ‘as expected, although there were some unexpected outcomes, most likely due to increased ethylene levels on board ISS.
The initial idea of having children on ground working together with an astronaut on board the ISS has proven to be a valuable concept to improve participation and affiliation with science and technology, at least during the DELTA mission. It would be interesting to know the long-term impact of ‘Seeds-in-Space’. Therefore the activity needs to be evaluated in about 5-10 years from now.
Acknowledgements: This work was supported by funds from the Dutch Ministry for Education, Culture and Science and NWO-SRON grant MG-057. Special thanks to the staff of our Russian partners in RKA-Energia and ESA staff and Dutch Space-DPO for supporting part of the hardware and tests.

DELTA-GPB : No abstract available.