amplify inflammation in humans as well as mice
Mortality from sepsis ranges between 30 and 50%
and is rising because of drug-resistant organisms,
a growing elderly population, and an increased
incidence of immunosuppression (25–28). The
failures of anti–Toll-like receptor 4, recombinant
activated protein C, and anti–TNF-a therapies in
clinical trials necessitate a rethinking of sepsis’ pathophysiology (6, 29–33). Because many early-phase
inflammatory cytokines operate concurrently and
redundantly, identifying upstream triggers may
generate therapies with broad downstream benefits. Altogether, the evidence shown here supports
the hypothesis that IL-3 mediates experimental
and human sepsis, is a major upstream orchestrator of the septic inflammatory phase, and can
be harnessed for therapeutic intervention.
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We thank M. Greene for secretarial assistance, M. Waring and
A. Chicoine for sorting cells, and K. Joyes for editing the
manuscript. The data presented in this manuscript are tabulated
in the main paper and in the supplementary materials. The
General Hospital Corporation has filed a patent application
(61/973,458) with the U.S. Patent and Trademark Office entitled
“Agents and Methods for Diagnosing and Treating Sepsis,” which
names F.K.S. and G.F. W. as inventors. Il3–/– mice are available
from Riken, Japan, under a materials transfer agreement.
This work was supported by NIH grants 5R01HL095612 and
R56-AI104695 and the Massachusetts General Hospital Howard
M. Goodman Fellowship (F.K.S.). G.F. W. was supported by the
German Research Foundation (WE4892/1-2 and 3-1). B.G.C. was
supported by the Société Française d’Anesthésie-Réanimation
(SFAR), Institut Servier, Fondation Groupe Pasteur Mutualité, and
Fulbright Scholarships (Monahan Foundation and Harvard French
Scholarship Fund). M.N. was supported by an Erwin Schrödinger
Fellowship of the Austrian Science Fund FWF (J3486-B13). The
authors declare no conflicts of interest.
Materials and Methods
Figs. S1 to S15
Tables S1 to S6
4 December 2014; accepted 21 January 2015
attenuates age-related cardiac
decline in Drosophila
Shubhroz Gill,1,2 Hiep D. Le,1 Girish C. Melkani,3 Satchidananda Panda1*
Circadian clocks orchestrate periods of rest or activity and feeding or fasting over
the course of a 24-hour day and maintain homeostasis. To assess whether a
consolidated 24-hour cycle of feeding and fasting can sustain health, we explored the
effect of time-restricted feeding (TRF; food access limited to daytime 12 hours every
day) on neural, peripheral, and cardiovascular physiology in Drosophila melanogaster.
We detected improved sleep, prevention of body weight gain, and deceleration of
cardiac aging under TRF, even when caloric intake and activity were unchanged. We
used temporal gene expression profiling and validation through classical genetics
to identify the TCP-1 ring complex (TRiC) chaperonin, the mitochondrial electron
transport chain complexes, and the circadian clock as pathways mediating the
benefits of TRF.
To determine whether a daily rhythm of eeding and fasting without reducing caloric intake can improve health metrics, we sub- jected a 2-week-old wild-type (WT) Oregon-R strain (table S1) of Drosophila melanogaster
adults to ad libitum feeding (ALF) or 12-hour
time-restricted feeding (TRF) of a standard cornmeal diet exclusively during daytime. At nighttime, the TRF cohorts were placed in vials with
1.1% agar to prevent desiccation (fig. S1A). The
daily food intake was equivalent in both groups,
although ALF flies consumed some of their food
during nighttime (Fig. 1A). Unlike ALF flies, the
TRF group did not gain body weight at 5 and
7 weeks of age (Fig. 1B). The ability to fly (flight
index) was slightly improved in the TRF group
(Fig. 1C). Although the total daily activity was
equivalent between both groups of flies (Fig. 1D),
the TRF flies were more active during daytime.
Sleep (defined as five consecutive minutes of
inactivity) (1) assessment revealed that flies on
TRF had less daytime sleep, but more nighttime
and more total sleep, than the ALF flies (Fig. 1E
and fig. S1).
Increase in sleep duration correlates with improved cardiac function (2). Therefore, by high-speed video imaging of ex vivo denervated hearts
bathed in artificial hemolymph (3), we measured the diameter of the beating Drosophila
heart at full relaxation and contraction and
the time interval between successive contractions to calculate cardiac function parameters
(Fig. 2A). At 3 weeks of age, the performance
of both ALF and TRF hearts was indistinguishable with equivalent heart period (HP), systolic
diameter (SD), systolic interval (SI), diastolic
diameter (DD), diastolic interval (DI), arrhythmia index (AI), and heart contractility, measured as fractional shortening (FS) (Fig. 2, B
to F; fig. S2; and movie S1). In the next 2 weeks,
the cardiac performance in ALF flies exhibited
characteristic age-dependent deterioration (4),
with increased SI, DI, HP, and AI and reduced
DD, SD, and FS. TRF flies showed smaller changes
in these cardiac performance parameters in both
genders (fig. S2).
We investigated whether a limited period of TRF
early or late in life could attenuate age-dependent
1Regulatory Biology Laboratory, The Salk Institute for
Biological Studies, La Jolla, CA 92037, USA. 2Division of
Biological Sciences, University of California San Diego, La
Jolla, CA 92037, USA. 3Departments of Biology and
Molecular Biology, and Heart Institutes, San Diego State
University, San Diego, CA 92182, USA.
*Corresponding author. E-mail: email@example.com
(G.C.M.); firstname.lastname@example.org (S.P.)