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Data nasterii:

05.07.1919 - Romania

Data mortii:

09.06.2013U.S.A

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GIULIANO DE MEDICI TOMBBasilica di San Lorenzo

Sagrestia Nuova Florence

MICHELANGELO DI

LUDOVICO BUONAROTTISIMONI

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The influence of light and darkness on

circadian rhythms and relatedphysiology and behavior through theSCN in humans

The oscillator genes and proteins involvedin the mammalian circadian oscillator

http://upload.wikimedia.org/wikipedia/commons/9/9d/SCN_mam.jpghttp://upload.wikimedia.org/wikipedia/commons/5/55/Circadian_rhythm_labeled.jpg

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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)

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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. nightMin. - 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

Mc Govern et al, 1977

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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):21407-12Fig. 1

http://www.pnas.org/content/106/50/21407/F1.expansion.html

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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 inAwere 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 inAwith respect to numbers of

CD11b/CD14-positive spleen cells from Blower right.

Keller M. et al.-Proc.Natl.Acad.Sci.USA,2009 dec.15; 106(50):21407-12Fig. 2

http://www.pnas.org/content/106/50/21407/F2.expansion.html

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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 isregulated 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 supernatantwere 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

http://www.pnas.org/content/106/50/21407/F3.expansion.html

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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 globalgene 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 fromAwere 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

http://www.pnas.org/content/106/50/21407/F4.expansion.html

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Correlating the circadian profile of the immunemechanisms with the therapeutic intervention could

represent a safe way to improve clinical efficiency.

Understanding the circadian profile of theimmunologic dynamics makes possible a correct

interpretation of the specific characteristics of severaldiseases.

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Correlation of several diseases with circadianrhythms perturbating conditions (i.e. shiftwork)proves that these conditions induce systemic

inflammation independently from classic- riscfactors (i.e. for cardiovascular disease).

Despite the huge amount of works all over the

world, it still remains a long way to go to themoment of their application in current clinicalpractice.