Influence of gravity changes induced by parabolic flight on cytokine production in mouse spleen

Dysregulation of the immune system has been a well-documented effect of human exposure to a microgravity environment during space flight. These effects have included altered cytokine production, reduced proliferative responses, altered signal transduction pathways and altered distribution of peripheral immune cells. Recent reports have documented immunologic studies performed in-flight. When tested during space flight, delayed-type hypersensitivity was reduced, indicating a dysregulation of cell-mediated immune function. However, the mechanisms by which this occurs remain unclear. The production of cytokines plays a critical role in the ability of a host to mount an immune response against an invading microorganism. In this study, the alteration of cytokine responses in mice following parabolic flight (PF) was investigated.     

Parabolic flight experiments on physiological data acquisition and processing technologies using small jet aircraft (MU300)

The parabolic aircraft flight provides a short low gravity environment for approximately 20 seconds, which may not be sufficient for a research on the physiological phenomenon induced by actual weightlessness in space. However, the method is still useful to reveal essential and characteristic feature of physiological signs, and is available for testing hardware and also training of crew member during altered gravity. This paper reports the summary of parabolic flight experiments recently conducted as a NASDA program (1990-1992). The program is providing opportunities in low gravity research with small jet aircraft for researchers and agencies. The flight experiments in the life science area have been conducted mostly focused on a physiological changes and basic methodology which may be effective under the altered gravity condition. In this study, the following research team, NASDA, Research Institute of Environmental Medicine, Nagoya University, Toyohashi University of Technology, Tokyo Metropolitan Hospital, Torey Research Center and JSUP were involved and coordinated for the research.     

How to estimate the cost of point-of-care CD4 testing in program settings: an example using the Alere Pima Analyzer in South Africa

Integrating POC CD4 testing technologies into HIV counseling and testing (HCT) programs may improve post-HIV testing linkage to care and treatment. As evaluations of these technologies in program settings continue, estimates of the costs of POC CD4 tests to the service provider will be needed and estimates have begun to be reported. Without a consistent and transparent methodology, estimates of the cost per CD4 test using POC technologies are likely to be difficult to compare and may lead to erroneous conclusions about costs and cost-effectiveness. This paper provides a step-by-step approach for estimating the cost per CD4 test from a provider''s perspective. As an example, the approach is applied to one specific POC technology, the Pima™ Analyzer. The costing approach is illustrated with data from a mobile HCT program in Gauteng Province of South Africa. For this program, the cost per test in 2010 was estimated at $23.76 (material costs = $8.70; labor cost per test = $7.33; and equipment, insurance, and daily quality control = $7.72). Labor and equipment costs can vary widely depending on how the program operates and the number of CD4 tests completed over time. Additional costs not included in the above analysis, for on-going training, supervision, and quality control, are likely to increase further the cost per test. The main contribution of this paper is to outline a methodology for estimating the costs of incorporating POC CD4 testing technologies into an HCT program. The details of the program setting matter significantly for the cost estimate, so that such details should be clearly documented to improve the consistency, transparency, and comparability of cost estimates.     

Evidence of short-term adaptation to microgravity of neuromuscular synergy during a whole body movement

Human, like any other animal systems, moves in the terrestrial gravity field and must learn gravity-related motor strategies during his ontogenetic development. Considering continuous gravity action upon body segments, movement involves particular muscle activation patterns depending on body orientation to gravity. Gravitational-altered environments provided by parabolic flight or orbital space mission offer a great opportunity to investigate how gravity is taken into account in posture and movement planning. Indeed, in a context where the mechanical constraints are modified, movement execution involves that new muscular activity have to be produced. Almost, only few studies in microgravity environment are included electromyographic analysis and this parameter is generally used only to confirm modification of the muscular activation patterns. This study is focused to analyse the adaptation capacity of the brain to a modified gravitational environment. In this aim, EMG activity have been recorded during a whole body movement execution in both normo- and microgravity environment during parabolic flight. This procedure allowed us to analyse the EMG patterns recorded during the very first moments of weightlessness. In this study are reported the results of this analyse.     

Gravity effects on cellulose assembly

The effect of microgravity on cellulose synthesis using the model system of Acetobacter xylinum was the subject of recent investigations using The National Aeronautics and Space Administration's Reduced Gravity Laboratory, a modified KC-135 aircraft designed to produce 20 sec of microgravity during the top of a parabolic dive. Approximately 40 parabolas were executed per mission, and a period of 2 x g was integral to the pullout phase of each parabola. Cellulose biosynthesis was initiated on agar surfaces, liquid growth medium, and buffered glucose during parabolic flight and terminated with 2.0% sodium azide or 50.0% ethanol. While careful ground and in-flight controls indicated normal, compact ribbons of microbial cellulose, data from five different flights consistently showed that during progression into the parabola regime, the cellulose ribbons became splayed. This observation suggests that some element of the parabola (the 20 sec microgravity phase, the 20 sec 2 x g phase, or a combination of both) was responsible for this effect. Presumably the cellulose I alpha crystalline polymorph normally is produced under strain, and the microgravity/hypergravity combination may relieve this stress to produce splayed ribbons. An in-flight video microscopy analysis of bacterial motions during a parabolic series demonstrated that the bacteria continue to synthesize cellulose during all phases of the parabolic series. Thus, the splaying may be a reflection of a more subtle alteration such as reduction of intermicrofibrillar hydrogen bonding. Long-term microgravity exposures during spaceflight will be necessary to fully understand the cellulose alterations from the short-term microgravity experiments.     

Considerations for non-invasive in-flight monitoring of astronaut immune status with potential use of MEMS and NEMS devices

The dynamics of how astronauts' immune systems respond to space flight have been studied extensively, but the complex process has not to date been thoroughly characterized, nor have the underlying principles of what causes the immune system to change in microgravity been fully determined. Statistically significant results regarding overall immunological effects in space have not yet been established due to the relatively limited amount of experimental data available, and are further complicated by the findings not showing systematically reproducible trends. Collecting in vivo data during flight without affecting the system being measured would increase understanding of the immune response process.The aims of this paper are to briefly review the current knowledge regarding how the immune system is altered in space flight; to present a group of candidate biomarkers that could be useful for in-flight monitoring and give an overview of the current methods used to measure these markers; and finally, to further establish the need and usefulness of incorporating real-time analytical techniques for in-flight assessment of astronaut health, emphasizing the potential application of MEMS/NEMS devices.     

Laboratory Evaluation of the Alere q Point-of-Care System for Early Infant HIV Diagnosis

Introduction

Early infant diagnosis (EID) and prompt linkage to care are critical to minimise the high morbidity and mortality associated with infant HIV infection. Attrition in the “EID cascade” is common; however, point-of-care (POC) EID assays with same-day result could facilitate prompt linkage of HIV-infected infant to treatment. Despite a number of POC EID assays in development, few have been independently evaluated and data on new technologies are urgently needed to inform policy.

Methods

We compared Alere q 1/2 Detect POC system laboratory test characteristics with the local standard of care (SOC), Roche CAP/CTM HIV-1 qualitative PCR in an independent laboratory-based evaluation in Cape Town, South Africa. Routinely EID samples collected between November 2013 and September 2014 were each tested by both SOC and POC systems. Repeat testing was done to troubleshoot any discrepancy between POC and SOC results.

Results

Overall, 1098 children with a median age of 47 days (IQR, 42–117) were included. Birth PCR (age <7 days) comprised of 8% (n = 92) tests while 56% (n = 620) of children tested as part of routine EID (ages 6–14 weeks). In the overall direct comparison, Alere q Detect achieved sensitivity of 95.5% (95% CI, 91.7–97.9%) and a specificity of 99.8% (95% CI, 99.1–100%). Following repeat testing of discordant samples and exclusion of any inconclusive results, the POC assay sensitivity and specificity were 96.9% (95% CI 93.4–98.9%) and 100% (lower 95% CI 98%) respectively. Among birth PCR tests the POC assay had slightly lower sensitivity (93.3% vs 96.5% in routine EID) and higher assay error rate (10% vs 5% in samples of older children, p = 0.04).

Conclusion

Our results indicate this POC assay performs well for EID in the laboratory. The high specificity and thus high positive predictive value would suggest a positive POC result may be adequate for immediate infant ART initiation. While POC testing for EID may have particular utility for birth testing at delivery facilities, the lower sensitivity and error rate requires further attention, as does field implementation of POC EID technologies in other clinical care settings.     

A Comparison of Two Recorders for Obtaining In-flight Heart Rate Data

Measurement of mental workload has been widely used for evaluation of aircraft design, mission analysis and assessment of pilot performance during flight operations. Heart rate is the psychophysiological measure that has been most frequently used for this purpose. The risk of interference with flight safety and pilot performance, as well as the generally constrained access to flights, make it difficult for researchers to collect in-flight heart rate data. Thus, this study was carried out to investigate whether small, non-intrusive sports recorders can be used for in-flight data collection for research purposes. Data was collected from real and simulated flights with student pilots using the Polar Team System sports recorder and the Vitaport II, a clinical and research recording device. Comparison of the data shows that in-flight heart rate data from the smaller and less intrusive sports recorder have a correlation of.981 with that from the clinical recorder, thus indicating that the sports recorder is reliable and cost-effective for obtaining heart rate data for many research situations.     

Prediction of human orthostatic tolerance by changes in arterial and venous hemodynamics in the microgravity environment

In this article, we intentionally present exclusively the results of our recent studies of arterial and venous hemodynamics as predictors of human orthostatic tolerance during space flight and after the return to Earth. The possibility of in-flight orthostatic tolerance prediction by arterial hemodynamic responses to the lower body negative pressure (LBNP) and venous hemodynamic changes in response to occlusion of the lower extremities is demonstrated. For the first time, three levels of cerebral blood flow deficits during the determination of orthostatic tolerance in the course of the LBNP test performed in microgravity. We offer quantitative arguments for the dependence of the cerebral blood flow deficit on the degree of tolerance of the LBNP test. Patterns of arterial hemodynamics during LBNP were successfully used to diagnose the actual orthostatic tolerance and to follow its trend during flight, which testifies to the possibility of predicting orthostatic tolerance changes in an individual cosmonaut during space flight. Occlusion plethysmography of the legs revealed three levels of response of the most informative venous parameters (capacity, distensibility, and rate of filling) of the lower extremities correlated to the severity of decrease in orthostatic tolerance.     

Energy Expenditure and Metabolic Changes of Free-Flying Migrating Northern Bald Ibis

Many migrating birds undertake extraordinary long flights. How birds are able to perform such endurance flights of over 100-hour durations is still poorly understood. We examined energy expenditure and physiological changes in Northern Bald Ibis Geronticus eremite during natural flights using birds trained to follow an ultra-light aircraft. Because these birds were tame, with foster parents, we were able to bleed them immediately prior to and after each flight. Flight duration was experimentally designed ranging between one and almost four hours continuous flights. Energy expenditure during flight was estimated using doubly-labelled-water while physiological properties were assessed through blood chemistry including plasma metabolites, enzymes, electrolytes, blood gases, and reactive oxygen compounds. Instantaneous energy expenditure decreased with flight duration, and the birds appeared to balance aerobic and anaerobic metabolism, using fat, carbohydrate and protein as fuel. This made flight both economic and tolerable. The observed effects resemble classical exercise adaptations that can limit duration of exercise while reducing energetic output. There were also in-flight benefits that enable power output variation from cruising to manoeuvring. These adaptations share characteristics with physiological processes that have facilitated other athletic feats in nature and might enable the extraordinary long flights of migratory birds as well.     

Evaluating Operational Specifications of Point-of-Care Diagnostic Tests: A Standardized Scorecard

The expansion of HIV antiretroviral therapy into decentralized rural settings will increasingly require simple point-of-care (POC) diagnostic tests that can be used without laboratory infrastructure and technical skills. New POC test devices are becoming available but decisions around which technologies to deploy may be biased without systematic assessment of their suitability for decentralized healthcare settings. To address this, we developed a standardized, quantitative scorecard tool to objectively evaluate the operational characteristics of POC diagnostic devices. The tool scores devices on a scale of 1–5 across 30 weighted characteristics such as ease of use, quality control, electrical requirements, shelf life, portability, cost and service, and provides a cumulative score that ranks products against a set of ideal POC characteristics. The scorecard was tested on 19 devices for POC CD4 T-lymphocyte cell counting, clinical chemistry or hematology testing. Single and multi-parameter devices were assessed in each of test categories. The scores across all devices ranged from 2.78 to 4.40 out of 5. The tool effectively ranked devices within each category (p<0.01) except the CD4 and multi-parameter hematology products. The tool also enabled comparison of different characteristics between products. Agreement across the four scorers for each product was high (intra-class correlation >0.80; p<0.001). Use of this tool enables the systematic evaluation of diagnostic tests to facilitate product selection and investment in appropriate technology. It is particularly relevant for countries and testing programs considering the adoption of new POC diagnostic tests.     

Lay testing cadres and point-of-care diagnostic tests for HIV and other diseases: An essential combination in health service delivery

Zibusiso Ndlovu and Tom Ellman discuss the potential value of task sharing in provision of testing for HIV and other infectious diseases.

Summary points

• Lack of access to testing plays a major role in the underdiagnosis of infectious and noncommunicable diseases, leading to higher morbidity and mortality.
• Point-of-care (POC) tests can offer rapid results, allowing for timely initiation of therapy. However, mere availability of POC tests in health facilities does not ensure utilization. Conducting POC tests has been shown to be a burden on highly trained frontline healthcare workers (HCWs; clinicians and nurses), who often have a broader scope of responsibilities and are critically scarce in many settings.
• The continual emergence of easy-to-use POC tests has not been accompanied by investment in a cadre of health workers to support their delivery, especially at decentralized health facilities where patients initially seek healthcare support.
• Historically, implementation of task shifting for POC tests has proven difficult due to lack of integration into national human resource structures and fiscal plans and lack of explicit national policies promoting task shifting, together with resistance from laboratory professionals.
• We propose that the scope of work for existing lay health worker (LHW) cadres could be broadened/remodeled to include POC tests for HIV services including for advanced HIV disease and other priority diseases, especially in primary healthcare or lower level facilities without laboratories. Policy makers and national program managers could ensure that this is part of broader national health workforce policies.
• Concerns of professional and/or regulatory bodies must be addressed, and these bodies (medical and laboratory councils) can guide national policy on which POC tests can be task shifted to less laboratory-trained cadres, and they could also lead in the development of a framework of education and supervision to ensure sustainability and maintenance of professional standards.
• Lay testing initiatives can scale up access to the multitude of available essential POC tests, for maximal impact of disease testing, closer to where people live. This can improve global health and accelerate progress toward universal health coverage.

     

Recording of blood pressure,heart rate and aortic nerve activity during parabolic flight in the rat via radio-telemetry

Exposure to microgravity induces cardiovascular deconditioning characterized by orthostatic hypotension when astronauts return to the earth. In order to understand the mechanism of cardiovascular deconditioning, it is necessary to clarify the changes in hemodynamics and the cardiovascular regulation system over the period of space flight. The telemetry system applied to freely moving animals will be a useful and appropriate technique for this kind of long term study of the cardiovascular system in the conscious animal during space flight. The purpose of the present study is twofold: firstly, to observe the detailed changes of arterial pressure and heart rate (HR) during microgravity elicited by the parabolic flight in order to study the acute effect of microgravity exposure on the cardiovascular system; and secondly, to test the feasibility of the telemetry system for recording blood pressure, HR and autonomic nervous activities continuously during space flight.     

Comparison of Venous and Capillary Differential Leukocyte Counts Using a Standard Hematology Analyzer and a Novel Microfluidic Impedance Cytometer

Capillary blood sampling has been identified as a potentially suitable technique for use in diagnostic testing of the full blood count (FBC) at the point-of-care (POC), for which a recent need has been highlighted. In this study we assess the accuracy of capillary blood counts and evaluate the potential of a miniaturized cytometer developed for POC testing. Differential leukocyte counts in the normal clinical range from fingerprick (capillary) and venous blood samples were measured and compared using a standard hematology analyzer. The accuracy of our novel microfluidic impedance cytometer (MIC) was then tested by comparing same-site measurements to those obtained with the standard analyzer. The concordance between measurements of fingerprick and venous blood samples using the standard hematology analyzer was high, with no clinically relevant differences observed between the mean differential leukocyte counts. Concordance data between the MIC and the standard analyzer on same-site measurements presented significantly lower leukocyte counts determined by the MIC. This systematic undercount was consistent across the measured (normal) concentration range, suggesting that an internal correction factor could be applied. Differential leukocyte counts obtained from fingerprick samples accurately reflect those from venous blood, which confirms the potential of capillary blood sampling for POC testing of the FBC. Furthermore, the MIC device demonstrated here presents a realistic technology for the future development of FBC and related tests for use at the site of patient care.     

Developmental testing of the Advanced Animal Habitat to determine compatibility with rats and mice

The Research Animal Holding Facility (RAHF) and the Animal Enclosure Module (AEM) have housed rats during Space Shuttle flights since the 1980s, but the operational constraints of the hardware have limited the scientific return from these Shuttle flights. The RAHF provides environmental control and monitoring for 24 rats with in-flight animal access, but it must be flown in the Spacelab. Due to the infrequent availability of Spacelab flights, rodent experiments rely heavily on the AEM. Unfortunately, the AEM supports only six rats, has no environmental control and provides no animal access in flight. The Advanced Animal Habitat (AAH) is being developed to support up to 12 adult rats or 30 adult mice for up to 30 days, provide active temperature control, animal telemetry and on-orbit video, record environmental parameters in the animal cage, and provide in-flight animal access in the Middeck, the Spacelab or the Space Station. To ensure the AAH can meet these requirements, animal testing is being conducted with rats and mice in every step of development. Testing began with the cage configuration.     

Development of a whole blood staining device for use during space shuttle flights.

BACKGROUND: Exposure to microgravity during space flight results in profound physiologic changes. Numerous studies have shown changes in circulating populations of peripheral blood immune cells immediately after space flight. It is currently unknown if these changes result from exposure to microgravity or are caused by the stress of reentry and readaptation to gravity. METHODS: We have developed the whole blood staining device (WBSD) as a system for the staining of whole blood collected during space flight for subsequent flow cytometric analysis. This device contains all liquids to address safety issues concerned with space flight and also moves the cells through the staining, lyse/fixation, and dilution steps. RESULTS: Data from flow cytometric analysis of samples stained in the WBSD was found to be comparable to data from samples stained by the conventional methods. Cells stained with the WBSD remain stable in the device for up to 14 days. The necessary manipulations required to use the device were tested on the KC-135 aircraft during the reduced gravity segment of parabolic flight. CONCLUSIONS: With the WBSD immunophenotype analysis can be performed at various time points for the duration of an entire Shuttle flight. In addition, this device has significant terrestrial applications for rapid and easy immunofluorescence labeling of whole blood in remote and isolated locations where immediate access to specialized equipment and skilled laboratory personnel may not be available. The WBSD provides a simple mechanism to design specific immunophenotyping tests for use by nontechnical personnel at bedside or in field locations. Cytometry 37:74-80, 1999. Published 1999 Wiley-Liss, Inc.     

Physiological problems of manned mission to Mars

Harsh environment and extreme factors related to the supposed exploration missions to Mars are considered as well as concomitant human organism reactions. Further investigations are required to get insight into the effects of gravity ranging from microgravity to hypogravity to hypergravity the crew will be exposed to during this voyage. A special emphasis should be placed on the studies of artificial gravity as an alternative to the existing in-flight countermeasures. Other issues to be attended include transitory states of human organism as a response to changes in gravity, effects of ionizing radiation and synergy of the variety of flight factors, and mechanisms of the hypomagnetic effects.     

Shift in arm-pointing movements during gravity changes produced by aircraft parabolic flight.

It has been shown that target-pointing arm movements without visual feedback shift downward in space microgravity and upward in centrifuge hypergravity. Under gravity changes in aircraft parabolic flight, however, arm movements have been reported shifting upward in hypergravity as well, but a downward shift under microgravity is contradicted. In order to explain this discrepancy, we reexamined the pointing movements using an experimental design which was different from prior ones. Arm-pointing movements were measured by goniometry around the shoulder joint of subjects with and without eyes closed or with a weight in the hand, during hyper- and microgravity in parabolic flight. Subjects were fastened securely to the seat with the neck fixed and the elbow maintained in an extended position, and the eyes were kept closed for a period of time before each episode of parabolic flight. Under these new conditions, the arm consistently shifted downward during microgravity and mostly upward during hypergravity, as expected. We concluded that arm-pointing deviation induced by parabolic flight could be also be valid for studying the mechanism underlying disorientation under varying gravity conditions.     

The mechanisms underlying neuromuscular changes in microgravity environment.

We reported that the levels of electromyogram in soleus muscle and the afferent neurogram recorded at L5 segmental level of the spinal cord were instantly decreased in response to exposure to microgravity (micro-G) environment created during a parabolic flight, although these activities were constantly presented at 1-G. It was also observed that the soleus muscle length was passively shortened in micro-G due to the plantarflexion of ankle joint. Similar phenomena were also induced by acute hindlimb suspension at 1-G. Further, the soleus muscle atrophied, if the dorsal root at L5 was transected. These results suggested that the unloading-related effects on muscle are closely associated with the inhibition of the afferent input. However, effects of gravity on most of the cells in the whole body can not be removed, although hindlimb suspension can inhibit the antigravity activity of rat hindlimb muscles. And parabolic flight can create micro-G only for a short period of time. Further, effects of hypergravity before and after micro-G are unavoidable. Therefore, further experiments utilizing space environment are essential.