Life Science Support for the NASA Space Radiation

Life Science Support for the NASA Space Radiation Laboratory (NSRL) Presented by Peter Guida

Life Science Support Long Term in Medical Building Animals Cells Short Term Staging at NSRL Animals Cells • From FY 14 on, all 4 of these facets have been handled by one team, thereby streamlining operations and saving money.

Animal-based Operations, Long Term • Fully AAALAC accredited since 1964 (up for next 3 year renewal in summer of 2017). Has never had a case of pathogen, parasite or virus transmission. • Can support ~8000 rodents per year as presently configured (space was reduced from 10, 367 sq. ft. to 2, 654 sq. ft. to begin FY 14). This was adequate for 2013 & 2014 due to reduced animal numbers. Some additional space was used, as needed, in 2015 & 2016.

Recent Husbandry of NASA Rodents Number of BLAF NASA Animals per Year 12000 10222 10000 # of Animals 8703 7629 8000 9127 8190 6368 6033 6000 4000 5402 3778 2000 0 2007 2008 2009 2010 2011 2012 2013 2014 2015 2016 • From 2007 to 2012, there was a fairly steady increase in the number of animals supported and utilized for NASA studies in the NSRL Program. Numbers fell in 2013 & 2014, then rebounded in 2015 & 2016).

The Ventilated Caging System in Medical: • Consists of 5 racks and a ventilation/filtration system. Was installed in 2009, then expanded in 2011 • Houses 240 -960 NASA mice in one room simultaneously (depending on 1 -4 mice per cage)

Animal Husbandry at NSRL • Includes 5 racks and a ventilation/filtration system in largest (250 sq. ft. ) room, and 2 smaller (175 sq. ft. ) rooms with conventional caging systems

Cell Culture Operations & Offices (in Medical) Tissue Culture Labs General Labs User & Support Team Offices Storage rooms • 16 cell culture labs (~4, 600 sq. ft. ). For FY 15, 1 was relinquished due to reduction in number of NSRL teams and as a cost savings measure. • 2 general labs (mouse perfusion, flow cytometry, RNA isolation, PCR, plants/seeds, bacteria, worms, yeast) • 16 user offices (computer w/internet, phone). One less starting FY 15.

Cell/Tissue Culture Laboratories in Medical • Each is ~250 sq. ft. in size, with required equipment typical to support these types of studies

NSRL Researcher Offices in Medical • Each is ~125 sq. ft. in size, with computer, router and telephone. Connects to the team’s laboratory

Cell Culture Operations at NSRL • Consists of 2 labs for cell/tissue culture operations and 1 lab for additional processing of samples, including 2 chemical fume hoods (each room ~325 sq. ft. ). They contain the laboratory equipment routinely needed by NSRL users.

The NSRL Life Sciences Support Team There was a reduction from 13. 47 FTE to 10. 8 to begin FY 15. This was increased to 11. 3 in FY 17. Scientific Support Staff Administrative Support Staff Peter Guida Technical Support Staff Laura Loudenslager Angela Kim Deborah Snyder Corrine Baran Kristie Bonti Bea Pyatt Jim Jardine Paula Bennett Bill Ward Leah Selva

Responsibilities: ~2+ Months Before Run • Read all users’ beam time proposals • Participate in IACUC (Institutional Animal Care and Use Committee), SACRR (Scientific Advisory Committee on Radiation Research) & Experimental Safety (ESR) reviews • Draft run schedule (in Statement of Work-SOW) • Create distribution list for run • Start weekly teleconferences w/users & distribute minutes • Review guest appointment requests • Schedule service contract work as needed • Begin readying labs (check operation of equipment, obtain certifications, schedule necessary work • Perform dry runs with new cell & animal teams

Responsibilities: ~1 Month Before Run • Create and finalize run schedule (SOW) • Evaluate and respond to GSRDs (Ground Support Requirements Documents) and animal information forms • Allocate lab/office space and equipment based on above • Read all proposals and hold Environmental Safety Reviews • Make information binders for users • Set up appropriate Radiological Work Permits • Contact users who require controlled substances • Continue to prepare labs (clean, sterilize >60 incubators, set up equipment in labs as per GSRD responses)

Responsibilities: ~2 Weeks Before Run • Perform sterility tests on all incubators • First shipments of users’ supplies (out of ~500 boxes) begin arriving. Unpack/label/store in assigned spaces at proper temperatures • Finish preparing labs (lab benches, water baths, Coulter Counters, etc. ) • Check office phones and voice mail systems • Check office computers • First users begin arriving

Responsibilities: During Run • Check users in (distribute keys, info. binders, give tour of building and assigned spaces, make after hours arrangements) • Continue to receive, unpack and store shipped goods • Provide transportation of cells (as needed) to and from NSRL • Provide transportation of all animals to and from NSRL • Monitor animals & rooms 7 days/week. Unpack/pack animals • Provide “on the spot” support for users • Assist with completion of forms and proper disposal of all medical, hazardous and radiological wastes • Hold daily run meeting with users to plan for upcoming needs • Long Term animal and cell facilities never close. Be on call and available 24/7 (~7 months per year)

Responsibilities: Daily at NSRL During Run • 7: 30 am: Cell culture facility startup – Confirm operational status and cleanliness of all cell culture, microscopy and beamline incubators (sterility, temperature, CO 2) and other laboratory, microscopy and computer equipment • 8: 30 am: NSRL daily user meeting – Confirm user needs are satisfied for conducting experiments, Q&A session • 9: 00 am– 5: 00 pm+: NSRL user support – Assist users with planning and conducting experiments, running samples in & out of target area as needed, dosimetry • End of day: Cell culture facility closedown – Replace/sterilize incubator components, clean/sterilize laboratory work surfaces, return used incubator components for cleaning/autoclaving

Responsibilities: After Run • Remove all wastes from labs for proper disposal (nonhazardous, medical and hazardous) • Shut down, empty and clean incubators • Clean labs • Send experimental samples and supplies back to users (150200 non-animal shipments per campaign) • Cross check inventories of remaining user supplies for next run • Organize and store supplies needed for next run

Summary of Life Sciences Support Positions: Administrative Assistant: take/distribute minutes, budgets, distribution lists, inventories, ordering, Summer School support Animal Technicians: provide husbandry & monitoring 7 days/week for >8000 NASA rodents/year, prep. animal rooms, transportation to/from NSRL, receive & send animal shipments Associates: prep. all 16 labs, receive/send hundreds of shipments of supplies/samples, create/maintain information databases, check users in/out, inventories, tend to “on the fly” needs/changes, all wastes. Assigned 7 -9 teams/campaign, so outnumbered >30: 1. Additional specifics for associates: • Loudenslager: anesthesia systems & monitoring, controlled substances • Kim: fluorescence microscope training/scheduling/troubleshooting • Pyatt: infection control practitioner (blood & primary human cell teams) • Bonti: cell counters, LN 2 & water purification system maintenance, QC & QA

Summary of Life Sciences Support Positions: • Scientist: Create run schedule, run daily user meetings & pre-run teleconferences, manage budgets, allocate resources, supervise staff, read all beam time proposals, serve as host for BNL guest appointments, act as Spokesperson for Experimental Safety Reviews, oversee use of animal subjects & work with Attending Veterinarian, maintain steady communication with researchers & answer questions, facilitate satisfaction of users’ training requirements, verify IACUC compliance, provide NSRL experimental & scientific support, act as interface with NSRL Physicists • NSRL Administrator: administrative support, preparation & communication of NASA user approvals, revisions and NSRL campaign calls, arrange SACRR reviews and teleconferences, maintenance of NSRL user databases, integration of & uploading to web-based system (PETRA) • Beamline Technicians: daily NSRL cell lab startups & shutdowns, lab equipment/incubator maintenance (cleaning & sterilizing), troubleshooting & certifications, instrument purchases/inventory tracking, mycoplasma testing, providing supplies (wet/dry ice, LN 2, etc. ), gamma and Tandem facility irradiations & support, and perform cave entries & computer/dosimetry support at NSRL

Representative Run Schedule Week:

Brookhaven Lab Animal Facility (BLAF) Issues: • With the rebound in animal numbers, and to have a steady presence at NSRL to support animal studies, Deborah Snyder was restored to full time (from 0. 5 FTE). This was done with funds from the Collider-Accelerator Department. • BLAF’s FY 17 operational cost is projected to be ~$800 k. NASA pays ~$350 k (44%), BNL the other 56%. The BNL Directorate continues to put $~450 k towards BLAF support each fiscal year. In FY 14 & FY 15, the BNL Directorate stated its reluctance about continuing this support going forward. However, funds were allocated in FY 16 and FY 17. • BLAF has stated that it would be highly beneficial to stretch out the duration of seasonal campaigns a little more in order to facilitate the receiving, housing, shuttling, and return shipping of animal subjects. This may not be possible.

Newest Important Issues/Items: • Costs: The Biosciences Department, which was merged with the Environmental & Climate Sciences Department as part of the Environment, Biology, Nuclear Science and Nonproliferation Directorate, was split off again and is now the separate Biology Department. The reduction in organizational burden and space charge rates from the previous merger has remained in place, thereby maintaining the cost savings. • Equipment: Several incubators from the former Sutherland team were claimed as NASA property and put into service in 2016 to replace malfunctioning units in the Medical building that were >10 years old. A cell culture hood was also claimed. These moves saved some of the money that was earmarked for new equipment going forward.

Newest Important Issues/Items: • GCR simulations - logistical and scheduling issues: Performed in NSRL 16 C, the first 3 GCRsims went off reasonably well. An important lesson learned was that the set up time needed for these types of studies is significant. Adam and I still think the best way to accommodate this is to have “mini runs” consisting of only GCRsim studies. • Rabbits as animal subjects: Following >9 months of MIRC and IACUC presentations, additional meetings and debate at BNL, the Boerma team successfully utilized rabbits as NSRL research study subjects. Husbandry, irradiations and shipping all went off without any significant issues. The final data points will be generated early in 2017.

Newest Important Issues/Items: • Experimental readiness: There has been a trend that more user teams are showing up to NSRL without adequate personnel and/or sufficient experimental information to properly direct their studies. • Logistical readiness: The number of teams needing addenda to their Institutional Animal Care and Use (IACUC) protocols at the last minute (i. e. the day before and even the day of scheduled beam time) has been steadily increasing. Nonconformances were avoided, but this leaves everyone vulnerable to a mishap. In the aftermath of the summer 2015 incident, the consequences of another non-conformance would be extremely dangerous.

Updates on Items from 2015: • Web site redesign: The NSRL web site, restructured and updated in 2015, received positive feedback from the NSRL research community and appeared to foster the use of the facility by non-NASA funded investigators. Adam, Mike Sivertz, Rory Rosselot and I have discussed the next wave of updates that should enhance the site further. • Low oxygen studies: Interest in performing these re-surfaced in the fall of 2016 as part of a new grant. The existing capabilities were utilized once again, but they will be expanded by making minor modifications to the present equipment.

Updates on Items from 2015: • Pharmacy situation: The DEA-sanctioned dispensary, created after BNL terminated the contract of its pharmacist, ran very well in 2016 with me in the role of “pharmacist”, with critical assistance from Laura Loudenslager. • Gamma facilities: All NASA users requiring gamma rays while at BNL have now used the J. L. Shepherd Mark I unit conveniently located in Medical. With excellent support from Jim Jardine, we have seen a steady stream of NSRL researchers using this source. • Fluorescence microscope: New computer and microscope camera (procured in 2015) continued to provide enhanced capabilities for NSRL researchers with outstanding efforts from Angela Kim.