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Impact of payload shielding on Enterobacter cloacae viability and proteomic profile: Insights from a stratospheric weather balloon flight experiment

Jonna Ocampo
Jonna Ocampo
, 
Hailey Barker
Hailey Barker
, 
Kelly C. Rice
Kelly C. Rice
 oraz 
Mariola J. Ferraro
Mariola J. Ferraro
   | 09 cze 2024
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Article Category: Research Note
Data publikacji: 09 cze 2024
Zakres stron: 64 - 76
DOI: https://doi.org/10.2478/gsr-2024-0005
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© 2024 Jonna Ocampo et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Figure 1.

An overview of the experimental design. (A) Timeline of the experiment. (B). Overall experimental plan.
An overview of the experimental design. (A) Timeline of the experiment. (B). Overall experimental plan.

Figure 2.

Payload configuration. A: Shielded (left) and unshielded (right) sample boxes. B: Sealed payload box.
Payload configuration. A: Shielded (left) and unshielded (right) sample boxes. B: Sealed payload box.

Figure 3.

Readings of temperature, pressure, and UV index from during the ascent and descent of the balloon flight. (A) External temperature measurements recorded during the flight. (B) Internal temperature measurements recorded during the flight. (C) Atmospheric pressure measurements taken throughout the flight. External UV index measurements from Sensor 1 (D) and Sensor 2 (E) during the flight.
Readings of temperature, pressure, and UV index from during the ascent and descent of the balloon flight. (A) External temperature measurements recorded during the flight. (B) Internal temperature measurements recorded during the flight. (C) Atmospheric pressure measurements taken throughout the flight. External UV index measurements from Sensor 1 (D) and Sensor 2 (E) during the flight.

Figure 4.

The intracellular survival of E. cloacae exposed to weather balloon flight in the payload shielded compared to unshielded conditions. E. cloacae exposed to the weather balloon flight in the presence or absence of Faraday fabric-based shielding were used to infect RAW 264.7 murine macrophages at an MOI of 80:1 for 2 hours post-infection (hpi) (A, C) or 24 hpi (B, D). After completion of the infection period, cells were lysed, and lysates were spread on LB plates to count CFUs. The experiment was repeated twice for a biological triplicate. For each individual experiment, average CFUs from infections with unshielded bacteria were considered as 100% (A, B), and one individual experiment is shown as total CFUs/well for each time point (C, D). Statistical significance was determined using a t-test. *, p < 0.05, **, p < 0.005, n.s. = not significant.
The intracellular survival of E. cloacae exposed to weather balloon flight in the payload shielded compared to unshielded conditions. E. cloacae exposed to the weather balloon flight in the presence or absence of Faraday fabric-based shielding were used to infect RAW 264.7 murine macrophages at an MOI of 80:1 for 2 hours post-infection (hpi) (A, C) or 24 hpi (B, D). After completion of the infection period, cells were lysed, and lysates were spread on LB plates to count CFUs. The experiment was repeated twice for a biological triplicate. For each individual experiment, average CFUs from infections with unshielded bacteria were considered as 100% (A, B), and one individual experiment is shown as total CFUs/well for each time point (C, D). Statistical significance was determined using a t-test. *, p < 0.05, **, p < 0.005, n.s. = not significant.

Figure 5.

The effect of E. cloacae exposure to weather balloon flight in the shielded compared to unshielded conditions on the TNF-α release from infected cells. E. cloacae exposed to the weather balloon flight in the presence or absence of Faraday fabric-based shielding as well as ground control were used to infect RAW 264.7 murine macrophages at an MOI of 80:1 for 2 hpi. After the infection time was complete, the cell culture supernatant was collected and the TNF-α was quantified in the media by using ELISA. N=4. For establishing statistical significance, a 1-way ANOVA test was used. *, p < 0.05, **, p < 0.005, ***, p < 0.0005, ****, p < 0.0001.
The effect of E. cloacae exposure to weather balloon flight in the shielded compared to unshielded conditions on the TNF-α release from infected cells. E. cloacae exposed to the weather balloon flight in the presence or absence of Faraday fabric-based shielding as well as ground control were used to infect RAW 264.7 murine macrophages at an MOI of 80:1 for 2 hpi. After the infection time was complete, the cell culture supernatant was collected and the TNF-α was quantified in the media by using ELISA. N=4. For establishing statistical significance, a 1-way ANOVA test was used. *, p < 0.05, **, p < 0.005, ***, p < 0.0005, ****, p < 0.0001.

Figure 6.

Proteomic analysis of the E. cloacae response to payload shielding during weather balloon flight. (A) Comparative proteomic profiling of E. cloacae exposed to shielded and unshielded conditions during weather balloon flight. The Venn diagram depicts the distribution of identified proteins, with 1217 proteins detected in both shielded and unshielded samples, 24 proteins unique to shielded conditions, and 114 proteins unique to unshielded conditions. Altered protein abundance analysis highlights 97 proteins with significant changes (p < 0.05, fold change > −1.5/+1.5); 22 proteins exhibited higher abundance in shielded samples, whereas 75 proteins were more abundant in unshielded samples. (B) Principal Component Analysis (PCA) showed distinct clustering of proteomic profiles between shielded and unshielded E. cloacae samples. Each data point represents a sample, with shielded and unshielded conditions forming separate groups based on protein expression patterns. (C) Unsupervised hierarchical clustering analysis of differentially regulated proteins in shielded and unshielded E. cloacae. The heat map reveals protein clusters exhibiting specific increases in abundance under each condition, allowing robust differentiation between shielded and unshielded samples.
Proteomic analysis of the E. cloacae response to payload shielding during weather balloon flight. (A) Comparative proteomic profiling of E. cloacae exposed to shielded and unshielded conditions during weather balloon flight. The Venn diagram depicts the distribution of identified proteins, with 1217 proteins detected in both shielded and unshielded samples, 24 proteins unique to shielded conditions, and 114 proteins unique to unshielded conditions. Altered protein abundance analysis highlights 97 proteins with significant changes (p < 0.05, fold change > −1.5/+1.5); 22 proteins exhibited higher abundance in shielded samples, whereas 75 proteins were more abundant in unshielded samples. (B) Principal Component Analysis (PCA) showed distinct clustering of proteomic profiles between shielded and unshielded E. cloacae samples. Each data point represents a sample, with shielded and unshielded conditions forming separate groups based on protein expression patterns. (C) Unsupervised hierarchical clustering analysis of differentially regulated proteins in shielded and unshielded E. cloacae. The heat map reveals protein clusters exhibiting specific increases in abundance under each condition, allowing robust differentiation between shielded and unshielded samples.

Figure 7.

Bioinformatic analysis of differentially abundant proteins in E. cloacae under weather balloon flight conditions with and without Faraday fabric-based shielding. (A) Enrichment analysis of Uniprot-assigned functions. (B) InterPro domain mapping identifies enriched protein domains. (C) STRING analysis.
Bioinformatic analysis of differentially abundant proteins in E. cloacae under weather balloon flight conditions with and without Faraday fabric-based shielding. (A) Enrichment analysis of Uniprot-assigned functions. (B) InterPro domain mapping identifies enriched protein domains. (C) STRING analysis.

Summary of Recorded Flight Data. This table presents a summary of the flight data (recorded by the primary flight computer), which lasted for a total of 89 minutes from launch to landing. The table highlights the minimum and maximum values recorded for altitude, external temperature, pressure, and UV index during this flight mission (for the entire dataset see Table S1).

Independent parameters: Minimum Maximum
Altitude 0 km 27 km
External temperature −57.8°C 25.04°C
External pressure 1.83 kPa 90.41 kPa
External UV index 1.44 19.13
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Język:
Angielski
Częstotliwość wydawania:
2 razy w roku
Dziedziny czasopisma:
Life Sciences, other, Materials Sciences, Physics
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