Data nasterii:

05.07.1919 - Romania
Data mortii:
09.06.2013 U.S.A
GIULIANO DE MEDICI TOMB
Basilica di San Lorenzo
Sagrestia Nuova Florence
MICHELANGELO DI
LUDOVICO BUONAROTTI
SIMONI
The influence of light and darkness on
circadian rhythms and related
physiology and behavior through the
SCN in humans
The oscillator genes and proteins involved
in the mammalian circadian oscillator
Limphocitar proliferation (Esquifino A . et al.,1996)
Natural-killer activity (Arjona A. et al., 2005)
Humoral immune response (Fernandes G.,et al. 1976)
Leukocitar population/subpopulation modulation
(Kawate T. et al.,1981)
Activated monocyte activity (Khoa D., et al., 2013)
Citocyne level (Young M. et al., 1995 ; Druzd
D.,Scheierman C., 2013)
Cortisolemia regulation (Krieger D., 1975; Herman N.,
2006 ; Kohsaka A., 2007)
Susceptibility to infection (Schakelford P., 1973)
Clinical entity Circadian manifestation References
Rheumatoid arthritis Maximum stiffness, pain ~8am Straub at al, 2007
Asthmatic attack Worst at night (mostly first
of the night + 4 - SA.M)
Gilbert, 1995; Martin et al,
1999
Ankilosing spondylitis Max. vertebral stiffness 6 - 8
A.M. and 6 9 P.M.
Focan Hensard D., 1988
Osteopolyarthosis Max. pain during
evening/night
Bellamy N., 1990; Deslandre
C., 1983; Levy E., 1985
Sepsis Max. mortality between
2A.M. and 8 A.M.
Hrushensky W., 1994
Psoriasis

epidermal cells
proliferation in affected areas

Dermic cells proliferation in
affected areas

Inflammatory activity


Max. 21 P.M. 3 A.M
Min. 9 A.M.

Max. 9 A.M.
Min. 3 A.M.

Max. night
Min. - morning



Rubin et al, 1983



Pigatto et al, 1985
Allergy/ allergy testing The skin is much less reactive
to testing early in the morning

Severity/identification of
specific sensitivity may be
misjudged if testing is
conducted early in the day


Mc Govern et al, 1977
Fully competent circadian clocks in tissues / cells of the immune system.
(Left) Circadian clock genes Per2 (filled circles) and Rev-Erb (open circles) are
rhythmically expressed in murine spleen cells, lymph nodes, and peritoneal
macrophages.
Tissues /cells were harvested at regular intervals over the the first 2 days
after transfer of the mice from a LD cycle to DD.
Gray and black bars refer to the previous light and
dark periods, respectively.
CT 0 corresponds to the time in DD when the light
would have turned on in the prior LD cycle.
Transcript levels were analyzed by using
quantitative RT-PCR (Right.).
A small piece of spleen, superficial inguinal lymph
nodes as well as peritoneal macrophages were
cultured in medium containing luciferin.
Circadian bioluminescence was continuously
recorded for 1 week by using photomultiplier tubes.
Representative time series for at least 3
independent experiments are shown.

Keller M. et al.-Proc.Natl.Acad.Sci.USA,2009 dec.15; 106(50):21407-12


Fig. 1
Circadian cytokine secretion upon challenge with bacterial
endotoxin.
(A) Spleens from C57BL/6 mice transferred in DD were
harvested at regular 4-h intervals.
After stimulation with LPS, TNF- (Left) and IL-6 (Right)
secretion was determined by ELISA.
Gray and black bars refer to the previous light and dark
periods, respectively.
(B) Cellular composition of the spleen is time-of-day
dependent.
The same samples as in A were analyzed with cell-
counting chamber and flow cytometry. CD19, CD90.2, and
CD11b in combination with CD14 were used as characteristic
surface markers of B cells, T cells, and monocytes/
macrophages, respectively.
(C) Cytokine response as in A with respect to numbers of
CD11b/CD14-positive spleen cells from B lower right.

Keller M. et al.-Proc.Natl.Acad.Sci.USA,2009 dec.15; 106(50):21407-12

Fig. 2
A macrophage intrinsic clockwork regulates circadian TNF- and IL-6
secretion upon LPS stimulation.
(A) Circadian modulation of LPS-induced cytokine response is
independent of systemic cortisol.
Spleens from adrenalectomized C57BL/6 mice were harvested and
analyzed as described in Fig. 2. TNF- and IL-6 cytokine secretion per
macrophage was determined via ELISA by taking the absolute number of
monocytes/ macrophages of the spleen into account .
(B) TNF- response upon LPS stimulation is
regulated by a cell-intrinsic, local clock. Spleen
cells from 20 C57BL/6 mice were harvested,
pooled, and plated for tissue culture.
Individual wells were stimulated for 4 h with
LPS at indicated times, and supernatants were
collected thereafter. TNF- levels in supernatant
were determined by ELISA and tested for
statistical significance.


Keller M. et al.-Proc.Natl.Acad.Sci.USA,2009 dec.15; 106(50):21407-12
Fig. 3
Fig. 4
8% of all transcripts in macrophages are expressed with a circadian rhythm.
(A) Phase-sorted heat map of genes transcribed in a circadian manner in
peritoneal macrophages. Cells harvested via peritoneal lavage from 4 C57BL/6
mice every 4h were magnetically purified for CD11b surface expression.
3 individual RNA samples of each time were pooled and subjected to global
gene transcription measurement The analysis on circadian rhythmicity was
done with CircWaveBatch. Genes expressed in a circadian manner were plotted
phase-sorted in a heat-map style (colors indicate minmax normalized relative
expression: green, minimum expression; red, maximum expression).
(B) Canonical clock gene expression in peritoneal macrophages. Individual
datasets from A were plotted (filled circles) together with data obtained by a
quantitative RT-PCR assay of the same samples (open circles)
Keller M. et al.-Proc.Natl.Acad.Sci.USA,2009 dec.15; 106(50):21407-12
Correlating the circadian profile of the immune
mechanisms with the therapeutic intervention could
represent a safe way to improve clinical efficiency.

Understanding the circadian profile of the
immunologic dynamics makes possible a correct
interpretation of the specific characteristics of several
diseases.


Correlation of several diseases with circadian
rhythms perturbating conditions (i.e. shiftwork)
proves that these conditions induce systemic
inflammation independently from classic- risc
factors (i.e. for cardiovascular disease).

Despite the huge amount of works all over the
world, it still remains a long way to go to the
moment of their application in current clinical
practice.