1692.pdf

T

a

b l

e o

f C

o n

t

e n

t

s

A bstr a ct 3

Te rm in olo_{g y} 4

Intr odu ctio n 6

A Ba ck gr o u nd 6

B A s s e s sin_{g A i}rwa_{y I}nfl ammatio n 7

C D if fe r en ces in Infl ammation atBas el ine 8

D O be sit_y a nd Infl ammatio n 10

E A ims 11

M

ethods 1 2

A Ge n e r al Sc r e e n ₍G S) Pr oto c ol 12

B G S Crite ria 12

C Ex clu sio n Crite ria fo r An al ysis 13

D Bod y

M

a s sInde x

(

B

M

I₎ 14

E G S Pr o c edu r e fo r Indu c ed S putum Pr o c e s sin_g 14

F Statistic al Anal ysis 15

1 E

_ff

e ct o

f

B

M

I o n Sputum Va riab le s 17

1 E

_ff

e ct o

f

D is e a s e Statu s o n Sputum Va ria_{b l}e s

f

o r D i

ff

e r e n tLe v els o

f

B

M

I 18

3 E

_ff

e ct o

f

A ge,Se x and Ra c e Co ntr ols o n Sputum Va riab le s 18

A Ac c o u ntin_g

_f

o r Lo_{g T}ran s

_f

o rmat io n s 19

Re s ults 2 0

A An a_lysis of Va ria n c e

(

A N O V A₎ 2 0

B

M

ultiv a riate L in e a r Re_gr e s sio n An a_lysis 2 3

\ E

_ff

e ct o

_f

B

M

I In c r e a s e o n Pri ma r_{y Sp}utum Va riab le s 2 4

2 E

ff

e ct o

_f

Dis e a s e Statu s o n Pri ma ry Sputum Va riab le s

_f

o r D i

_ff

e r e ntLe v els o

f

B

M

I 2 4

3 E

_ff

e ct o

f

A ge, Se x and Ra c e Co ntr ols o n Prima ry Sputum Va riab le s 2 8

D is c u s sio n 2 9

A A N O V A 2 9

B

M

ultiv a riate L in e a r Re_gr e s sio n An alysis 3 0

1 E

_ff

e ct o

_f

B

M

I In c r e a s e o n _Sputum Va riab le s 3 0

2 E

_ff

e ct o

_f

D is e a s e Statu s o n _{S p}u tum Va riab le s

_f

o r a G iv e n Le v elo

f

B

M

I 3 1

3 Sum ma r_y o

_f

Inte r a ctio n Bet we e n B

M

I a n d D is e a s e Sta tu s 3 2

4 E

_ff

e ct o

_f

_{A g}e, Se x and Ra c e Co ntr ols o n Sputum Va riab le s 3 2

C Str e n_gths a nd

W

e akn e s s e s 3 3

Co n clu sio n 3 4

Ackn ow led geme nts 3 5

Refe r e n c e s 3 6

A b

s

t

r a c

t

In

fl

a

m m

atio n

i

s a cha r a cteri stic fe atu r e o

f

air

w

a_y

d

is e a s e s s u ch a s ato_{p y} a n

d

a st

h m

a T

h

e

pr e s e n c e of n e utr o_{p h i l}s,

m

a c r op hage s a n

d

e o sin op h ils at

b

a s el in e in the air

w

ays i

n

_d

ic ate s

a ctiv atio n of the in n ate i

m m

u n e s_yste

m

O

b

e sit_y, cla s s

i f

i e

d b

y Bo

d

y

M

a s s In

d

e x

(

B

M

I

)

,

is

b

el ie v e

d

to

b

e l inke

d

w it

h

a state o

f

chr o n

ic

i

n

fl

a

m m

atio n T he

C

e nte r fo r

En vir o n

m

e ntal

M

e

d

ic

i

n e, Asth ma a n

d

Lu ng B io

l

og y

(

C

E

M

A L B

)

ha s a

da

ta

b

a s e

(

G

e n e r al

Sc r e e n

₍

G

S

₎

data

b

a s e

)

w

h ich ha s a n a

l

_yz e

d

s e v eral

h

u n

d

re

d

hu

m

a n v olu nte e r s '

s_putu

m

s am_p

l

e s

f

o r c e

l lu

l

a r

m

a r

k

e r s o

f in

fl

a

m m

at

io n

T

h

e s e

m

a r

k

e r s o r v a r

i

a

b l

e s, a

l l m

e a s u r e

d

d

uri n_g t

h

e

b

a s el in e state, in c

lu

d

e tota

l

a n

d di f f

ere ntia

l i

n

fl

a

m m

ato ry c e

l l

c o u nt T

h i

s

stu

d

_{y p}e rfo r

m

e

d

a o n e-w

a_{y A}n al ysis of V ari a n c e

(

A N O V A

)

a n

d

a

m

ultiv ari ate l in e a r

r e_gr e s s

i

o n a n a

l

_y sis o n the

G

S

da

ta

b

a s e v a riab le s to in v e st_{i g}ate the r elativ e

d

i

f

fe r e n c e s

in

b

a s e

l i

n e

l

e v e

l

s o

f i

n

fl

a

m m

at

i

o n amo n_g

h

e a

l

th_y, atop

ic

a n

d

a st

hm

at

i

c

in

d i

v

i d

u a

ls

, a n

d

to

a s s e s s the e

f

fe ct o

f

B

M

I o n in

fl

a

m m

atio n a s

m

o

de r a

te

d b

_y

di

s e a s e statu s, r e spe ctiv el y

A sth

m

atic in

d

iv

i

du als, o n a v e r age, ha

d

s

i

gn

i

fi c a nt

l

y fe

w

e r n u

m b

e r s o

f

a

ir

w

ay

m

a c r o_p

h

a_ge s tha n he a

l

th_{y i}n

di

v

i du a

ls a n

d

a _gr e ate r _pe r c e nta_ge o

f

e o s

i

n o_{p h}

i

ls t

ha n

ato_p

ic

a n

d

he a

l

th y in

_{d i}

v

_id

u a

ls

(p

<0 0 5

)

T he r e

w

a s a s

i

_gn

_{i fi}

c a n t _po s

i

t

iv e

inte r a ctio n

b

etwe e n

B

M

I a n

d

a sthmatic

in

di

v

i d

u a

l

s

fo r

the _pe r c e nta_ge o

f

n e utr o_p

h i l

s

(p

= 0 0 1 2 2

)

a n

d

a

c o r r e spo n

di

n_g n e_gativ e

i

nte r a ctio n fo r the _pe r c e nta_ge o

f m

a c r o_p

h

a_ge s

(p

=0 0 19 5

)

T he r e

w

e r e n o s

i

_gni

f

i c a nt

i

nte r a ct

io n s

b

et

w

e e n B

M

I a n

d

ato_p

ic

i

n

_di

v

i d

u a

l

s

O

v e r a

l l,

b

a s e

l i

n e

le v e

l

s of in

fl

a

m m

atio n

w

e r e s

i

_gni

f

i c a nt

l

_y

d

i

ff

e r e nt a

m

o n_g

h

e alt_{h y}, atop ic a n

d

a sth matic

in

_d

iv

i d

u als B

M

I a_{p p}e a r e

d

to

h

a v e a n ef fe c t o n c o n stitutiv e in

fl

a

m m

atio n

in

in

d

ivi

d

u als

T

e r

m

i

no

l

o

_{g y}

• A b_{s o}lu te

N

um ber o

_f

Cel ls_pe r m i l l igr am Total n u

m b

e r of a s_pe c

i f

i c c e

l

l t_{y p}e _pe r

m

_{i l l i g}r a

m

o

f

s ele ct s_putu

m

o A

b

s olute

N

um

b

e r o

f

Eo sin o_{p h i}

ls

_pe r m i

l

l igr am Tota

l

n u

m

be r of e o sin o_{p h i l}s _pe r

m

i

l li

_gr a

m

o

f

s e

l

e ct s_putu

m

o A bs olute

N

um

b

e r o

f

M

a c r o_{p h}a_ge s _pe r _{m i l l ig}r am Total n u

m b

e r of

m

a c r o_{p h}a_ge s

pe r

m

i l l i gr a

m

of s e

l

e ct s_putu

m

o A bs olute

N

um be r o

_f

N

e utr o_{p h i l}s _pe r m i l l i gr am Total n u

m

be r of n e utr o_{p h i l}s _pe r

m i l l i

_gr a

m

o

f

s e

l

e ct s_putu

m

• _Bod y

M

a s s Inde x

(

B

MI

)

: A

w

e

i

_{g h}t -fo r-h

ei g

h

t

i

n

d

e x u s e

d

to cla s si f y u n

d

e r

w

ei g

h

t,

n o r

m

a

l

, o v e r

w

e

i

g

h

t, a n

d

o

be s e a

d

u

lt

s

in

the Inte

m

atio n al

C l

a s si

fi

c atio n gu

i d

e

l in e s o

f

t

h

e

W

o r

l d H

e a

l

th Or_ga n

i

zat

io n

''*

It

i

s c alc u

la

te

d b

_y

d i

v

i di

n_g

w

e

i

_g

h

t

i

n

k i l

o_gr a

m

s

b

_y

h

ei g

ht

in

m

ete r s s_qu a r e

d

•

F

o r c e

d

Ex_{p i}r a to r_{y V}o

l

ume

(

F E V

)

:Total v o

lu

m

e of air that c a n

b

e r ele a s ed du r

i

n_g

e x

h

alat

i

o n

• _F o r c e

d

Ex_{p i}r ato r_{y V}o

l

ume in o n e s e c o n

d

(

F E V l

)

:T ota

l

v o

l

u

m

e o

f

a

i

r that c a n

b

e

r e

le a s e

d in the fi r st s e c o n

d

of e xhalatio n

• _Initia

_l

_Sputum

We

_{i g ht}

_{(m g}

₎

_:Tota

_l

_a

_m

_{o u nt o}

_f

_{e x}

pe cto r ate s_putu

m f

ol lo

w in

_{g i}n

d

u ct

io n

•

M

etha chol in e C hal le n_ge Te st

in

d

u cin_g

_b

r o n cho c o n stri ctio n in s u

b

j

e cts

b

_y

i

nhalat

i

o n

o

f

n e

b

u

l i

z e

d m

et

h

a c

h

o

l i

n e to

di

a_gn o s e a st

h m

a A po s

it

i

v e

m

et

h

a c

h

o

l i

n e c

h

a

l le n

_ge

in

di

c ate s a dr o_p of 2 0

%

o r

m

o r e in fo r c ed e x _{p i}r ato r_y v olu

m

e in o n e s e c o nd

(

F E V l

)

• P_{e r c e nt o}

_f

_Ce

_l

l Ty p_{e : Pr o}

po rtio n o

f

e a ch c e

l l

t_{y p}e in t

h

e total c e

l l

c o u nt o

f

s ele ct

o Pe r c e n tEo sin o_p

h

i ls :Pr o_po rtio n of e o s

in o

_{p h i l}s in the total c el l c o u nt of s ele ct

s_putu

m

o Pe r c e n t

M

a c r o_{p h}a_ge s :Pr o_po rtio n of

m

a c r o_{p h}a_ge s in the total c el l c o u nt of s ele ct

s_putu

m

o Pe r c e nt

N

e utr o_{p h i l}s :Pr o_po rtio n of n e utr o_{p h i l}s in the total c el l c o u nt of s ele ct

s_putu

m

• _Pe r c e nt Cel l V iab i l it_y: Pr o_po rtio n o

f

l iv e c el ls in t

h

e tota

l

c el l c o u nt of s ele ct s_putu

m

.

• Sele ct _Sputum

W

ei g

h

t

_{(m g}

)

: Total a

m

o u n t o

f m

a n u a

l l

_y s ele cte

d

s_putu

m

_p

l

u_g

m

ate rial

•

_S

p ir ometr_y :Pul

m

o n a r_y

f

u n ctio n te st

(

P F T

)

m

e a s u rin_g the v o

l

u

m

e o

f

in

h

a

l

e

d

o r

e xhale

d

air a s a

f

u n ct

io n o

f

ti me to e v alu ate the

d

e_gr e e of a

i

rwa_y o

b

str u ctio n in t

h

e

lu n_gs

•

_S

putum: Ex_pe cto r ate s e c r etio n s

fo

l l

ow in_g

i

n

d

u ct

io n

f

r o

m

t

h

e s u r

f

a c e s o

f

t

h

e c e ntr a

l

a

i

r

w

a_ys.

•

_S

putum P

l

u_gs : Po rtio n s o

f

s_putu

m

that a r e

d

e n s e r t

h

a n s u r r o u n

di

n_g s al iv a a n

d

t_{y p}

i

c a

l l

_y r

i

c

h

in c e

l l

u

la r c o n

te nt

I

n

t

r o

d

u c

t

io n

A

.

B

a c

k

g

r o u n

d

Infl a

m m

atio n is a c

h

a r a cte

ri

st

i

c fe atu r e o

f m

a n_y air

w

a_y

d

is e a s e s A st

h m

a is

cha r a cteri z e

d b

_y a state o

f

chr o n

i

c a

ir

w

a_y

i

nfl a

m m

atio n ^

'

^

w h ich s e v e r al stud ie s ha v e

i d

e nti

fi

e

d

a s a _p

ri m

a r_y c a u s e of the _patho_ge n e sis of the

d

is e a s e*

"_^

A ir

w

a_{y i}nfl a

m m

atio n

is a

ls o

b

el ie v e

d

to

h

a v e a vital r ole in al le r_{g i}c rh initis, o r atop y ^' ^' *

"_^

,tri g ge r e

d

in

r e spo n s e to a n al le r_ge n

A ir

w

a_{y i}n

fl

a

m m

at

io n

in v olv e s the in

fi

ltr atio n of infl a

m

mato r_y c el ls

f

r o

m

the

iim

ate a n

d

t

h

e a

d

a_ptiv e

i m m

u n e s_yste

m

T

h

e

i

n n ate

i m m

u n e c el

l

u

l

a r r e s_po n s e _pr o v

i d

e s a n

i m m

e

d

iate, n o n

-s_pe ci fi c

de

fe n s e a_gain st r e c o_gniz e

d

e ntit

i

e s o r _patho_ge n s

b

_{y i}n

fi

ltr atin_g at

t

h

e site of in

j

u r_y^ " T he in n ate r e s_po n s e c a n sti

m

ulate the slo we r, a nti ge n -_s

pe c

i fi

c

a

d

a_pt

i

v e i

m m

u n e c e

l l

ula r r e s_po n s e that

h

a s

i m m

u n o

l

o_g

i

c a

l m

e

m

o r_y to e

l i m i

n ate

r e c o_gniz e

d

e ntitie s o r _patho_ge n s

" ^

T

h

e

in n a

te

i m m

u n e s_yste

m i

s o

f

_gr ow

i

n_{g i}nte r e st in

air

w

a_y

d

is e a s e s, s u ch a s a sth

m

a a n

d

atop y, w

h

ich

in v o

lv e a

l l

e rg

ic r e s

po n s e s to ot

h

e rw is e

irmo c u o u s s u

b

sta n c e s' * In the s e

d

is e a s e s, the in n ate c el lula r r e spo n s e

m

istake nl y

r e c o_gn

i

z e s

m

o stl y

m

n o c u o u s e ntitie s a s_pat

h

o_ge n s a n

d

st

i m

ulate s a n

i m m

u n e r e s_po n s e

T he

fi

r st r e s_po n

d

e r s of the in n ate

i m m

u n e r e s_po n s e a r e _ge n e r a

l l

_y

m

a c r o_{p h}a_ge s

a nd n e utr o_{p h i l}s

M

a c r o_{p h}a_ge s a r e v e r s ati le _{p h}a_go c_ytic c el ls r e si

d

in _{g i}n la r_ge n u

m b

ers in

t

i

s s u e s A s t

h

e_y

m

atu r e

f

r o

m

c

i

r c u

l

a

ti

n_g

m

o n o c_yte s a n

d

a r e a ct

iv a

te

d

t

h

r o u_g

h

a n

ti

_ge n

r e c o_gn

i ti

o n,

m

a c r op

h

age s c a n e ngul f r e c ogniz e

d

e n

ti ti

e s a n d e n

do

ge n o u s

d

e

b ri

s a n

d

s e c r ete _pr o

-infl a

m m

ato r_y c_ytok in e s a n

d

othe r si gn al in_{g p}r ote

in

m

o

l

e c ule s to r e c r u

i

t

a

d di ti

o n a

l i

n

fl

a

m m

ato r_y c e

l l

s, s u c

h

a s n e utr op

h i l

s, to t

h

e s

i

te o

f i

n

j

u ry ''

the

m

o st c o

m m

o n t_{y p}e of gr a n u

l

o c_yte a n

d

cir c ulate in la r_ge n u

m b

e r s in the

b

lo od^ ^

Fol

l

o

w

in_g che

m i

c a

l

s

i

_gn a

l i

n_g, t

h

ey a r e r e c r u

i

te

d

to a n

d

a c c u

m

ulate at s

i

te s o

f i

n

j

u ry

w h

e r e the_{y p}e rfo r

m

_{p h}a_go c_{y t}ic fu n ctio n s a n d de_gr an ulate to rele a s e _pr ote a s e s' ^.

Eo sin o_{p h}

i l

s a r e le s s c o

m m

o n _gr an ulo c_yte s

l

o c ate

d m

o st_{l y i}n c o n n e ct

i

v e t

i

s s u e t

h

at

pri

m

a r_{i l y} ta r_get la r_ge _pa r a site s a n

d

ha v e s e c o nda r_{y p h}a_go c_ytic fu n ctio n'" Eo sin o_{p h i l}s

c a n r e

l

e a s e le u

k

otr

i

e n e s, e n zy

m

e s, cyto

k i

n e s, a n

d

ot

h

e r c

h

e

m

ic a

l m

e

di

ato r s t

h

at c a n

d

a

m

a_ge s u r r o u n

d i

n_g t

i

s s u e a n

d

r e c r u

i

t

m

o r e e o s

i

n o_p

h i l

s to t

h

e s

i

te o

f i

n

_j

u r_y'" T

h

e

pr e v ale n c e of e o sin o_{p h i l}s at site s of al

l

e r_{g i}c in

f

la

m m

atio n ind

i

c ate s that the_{y h}a v e a r ole

i

n c

h

r o n

i

c

i

n

f l

a

m m

at

i

o n

i

n

d i

s e a s e s s u c

h

a s a st

h m

a a n

d

ato_{p y}'^ Eo s

i

n o

_{p h}

i ls a r e _ge n e r a

l l

_y

n ot

f

o u n

d

in n o n

-ato_p

i

c a n

d

n o n

-a st

h m

at

i

c in

d i

vi

d

u a

l

s

B

.

A

s s e s s

i

n

g

A

i

r

w

a

y I

n

f

l

a

m

m

at

i

o n

Ex am

i

n

i

n_{g i}n

f l

a

m m

ato r_y

m

a r

k

e r s s u c

h

a s n e utr o_p

hi l

s,

m

a c r op hage s, a n

d

e o s

i

n o_{p h}

i

ls c a n

h

e_{l p} elu ci

d

ate u nde r_{l y i}n_g

m

e c

h

a nis

m

s o

f

o

b

str u ctiv e air

w

a_y

d

is e a s e at

ba s el in e in ind ivi du als w ith a sth

m

a a n

d

ato_{p y T h}e r e

l

ativ e _pr e s e n c e of inf la

m m

ato r_y

m

a r

k

e r s at

b

a s e

l

in e c a n a

l

s o

h

el p e

l

u ci

d

ate

m

e cha n

i

s

m

s o

f

the in n ate

i m m

u n e r e s_po n s e in

the s e in

d i

vi

d

u als

H i

sto ric a

l

_{l y}

,

b

r o n c

h

o s c op ic

m

etho

d

s

(

Br o n c

h

o alv e ola r La v age,

b

r o n c

h i

a

l

w

a s

h i

n_gs

,

b

iopsie s

)

h

a v e

b

e e n u s e

d

to e x a

m

in e the degr e e of air

w

ay in

f

la

m m

atio n in t

h

e

l

u n_gs. In t

h

e pa st twe nty ye a r s, howe v e r, in

d

u c e

d

sputu

m h

a s e

m

e rge

d

a s a n e

ff

e ct

i

v e a n

d

l

e s s

i

n v a siv e

m

et

h

o

d

to a s s e s s air

w

a_{y i}n

f

la

m

m

atio n In

d

u c e

d

s_putu

m

c o nte nt

i

s o

b

tain e

d

f

r o

m

the

l

ow e r r e s_{p i}r ato r_y tr a ct*' ' ^

thr o u_g

h

a r elativ e_{l y} n o n-i

n v a siv e

m

etho

d

in

c o

m

_pa r

i

s o n to

b

r o n c

h

o s c o_p

i

e s ^' *

"_' ' '

'*"_'

^

S

e v e r a

l

stu

d i

e s

h

a v e n ote

d

t

h

at, w

h

e n o

b

ta

i

n e

d i

n

a sta n

d

a r

d

ize

d m

a n n e r,

i

n

d

u c e

d

sputu

m i

s r epr o

d

u ci

b l

e

^' * "_^

' ' '*"'_*

infl a

m m

ato r_y c e

l

l c o nte nt o

b

tain e

d i

n in

d

u c e

d

s_putu

m

is c o

m

_pa r able to that of

b

r o n cho s c o_p

i

e s^' '^' "

S

pa n e v el lo et al ^ a n

d

Ba c ci e tal ' ^ha v e s_pe ci

fi

c a

l

_{l y}

d

e

m

o n str ate

d

go o

d

r e_pr o

d

u ci

b i l

it_y o

f

n e utr o_{p h i}

l

s,

m

a c r op hage s, a n

d

e o sin op h i ls

i

n sputu

m

c el l c o u nts

C

.

D

i f f

ere n c e s

i

n

I

n

f l

a

m m

at

i

o n at

B

a se

l in e

N

ichola s a n

d

D

j

u

k

a n o vic fo u n

d

that

i

n

du c e

d

s_putu

m is

_pote ntial l y u s eful in

in v e sti gatin_g

d i

s e a s e etio

l

o_{g y}

d

u e to

i

ts r

i

c

h

in

fl

a

m m

ato r_y c o nte nt' '_*

Si

m

i

la r

_{l y}, P in et a

l

.

d

e

m

o n str ate

d

that in

d

u c e

d

s_putu

m m

a_y

b

e u s eful in stu

d

_{y i}n_g a sth

m

a a n

d

othe r air

w

a_y

di

s e a s e s' Eo s

i

n o_{p h i}

l i

c in

fi l

tr atio n ha s

b

e e n

_{de s c r}

i

b

e

d

a s a cha r a cte r

is

tic fe atu r e

i

n

a sth ma'' ''' ' '^ ■^_''

A n u

m b

e r o

f

stu

di

e s

h

a v e s

h

o v

m

_gr e ate r e o sin o_p

h

i l ic

i

n

fl

a

m m

at

i

o n

in

1 1 © 0 *7Q 0 1

s u

b

j

e cts

w

ith a sth

m

a c o

m

_pa r e

d

to he a

lt

_{h y} c

hi

l

d

r e n '

a n

d

he alth_y a

d

ults ' "

'

Eo sin o_{p h i}

l

ic infl a

m m

atio n ha s

b

e e n r e_po rte

d

in ato_{p y} , a n

d

it

h

a s

b

e e n n ote

d

t

h

at

r e c u r r e nt a

l l

e r_ge n e x_po s u r e

h

elps s u stain e o sin o_p

h

i l in

fi l

tr atio n ^

To

d

ate, o nl y a

f

ew

stud ie s

h

a v e in v e st_{i g}ate

d

in

fl

a

m m

at

i

o n in ato_{p i}c s u

b

j

e cts,

b

ut w it

h

c o nfl ict

in

g r e s ults

A lv a r e z et al ^ a n

d S

_pa n e v el lo etal *

b

oth sho

w

ed t

h

at ato_{p i}c s u

b

_j

e cts ha

d

a _gr e ate r

pe r c e nta_ge o

f

e o s

i

n o_p

h i l

s r e

l

at

iv e

to

h

e a

l

th_y s u

b

j

e c_{t s}

b

ut a

l

owe r _pe r c e nta_ge c o

m

_pa r e

d

to

a sth mat

i

c s;

h

o

w

e v e r, A

l

v a r e z eta

l f

o u n

d

a gr e ate r pe r c e ntage o

f

e o s

in o

p

h i l

s

in

atop ic

s u

b

j

e cts c o

m

_pa r e

d

to

h

e a

lt

h y s u

b

j

e cts, w

h

i

l

e Spa n e v el lo et a

l

sho

w

e

d

n o

d i f

fe r e n c e in

the _pe r c e nta_ge of e o sin o_{p h i l}s bet

w

e e n ato_{p i}c a n

d

he a

lt

h y s ub

_j

e cts *

I

n c o ntr a st to a sth mat

i

c a n

d

ato_p

i

c

i

n

di

v

i d

u a

ls

,

h

e a

lt

h

y c

h i l dr e n

'

'

^

a n

d h

e alth_y

a

du

l

ts *' ^

h

a v e a r elat

iv e s c a r c

it

_y o

f

e o s

in o

_p

h i l

s ^

' *

a n

d

a _pr e

do

m i

n a n c e o

f m

a c r o_{p h}a_ge s

'' ' '

a n

d

n e utr o_{p h i l}s '' *

in t

h

e air

w

a_ys P in eta

l f

o u n

d

a si gni fi c a nt_{l y}

lo

we r _pe r c e nta_ge

o

f m

a c r o_{p h}a_ge s a n

d

a tr e n

d fo r a

l

o

w

e r _pe r c e nta_ge o

f

n e utr o_p

h

i ls

i

n a sth

m

atic s u

b

j

e cts

w h

e n c o

m

_pa r e

d

to he alth_y s u

b

j

e cts a n

d

ato_{p i}c s u

b

j

e cts, r e spe ctiv el y '

Jataka n o n et al n ote

d

a n ele v ate

d

n u

m b

e r o

f

n e utr o_{p h i l}s

i

n s e v e r e a sth

m

atic s c o

m

_pa r e

d

to

m

il

d

a sth

m

atic s a nd he alth y s ub

_j

e cts, ind ic ating a role o

f

n e utr op h i ls in s e v ere a sth ma

o r in a st

h m

a e x a c e rbatio n s ^

^

L i

k

e

w

is e, Fah y

d

e s c ri be

d

n e utr op h i ls a s the

m

o st abu nda nt

c el l t_{y p}e in

i

n

d

u c e

d

s_putu

m

in

b

oth

h

e alt

h

_y a n

d

a sth

m

atic s u

b

j

e cts^ '

H

o

w

e v e r, Fo r e si et

al r e_po rted n o si gn i fic a n t d i f fere n c e in the _perc e nta_ge of n e utro_{p h i l}s a n d

m

a c r o_{p h}a_ge s

b

et

w

e e n a sth

m

atic a n

d

ato_{p i}c s u

b

j

e cts ^

A ltho u_{g h}

m

a c r o_{p h}a_ge s a n

d

n e utr o_p

h i

ls a r e _pr e v a

l

e nt

i

n s_putu

m

o

f h

e alth_y

s ub

j

e cts, a pr edo

m

in a n c e of n e utr op h i

l

s in sputu

m

of a sth

m

atic s ub

j

e cts ha s als o be e n

o

b

s e r v e

d

T he r e

i

s a

l

s o a

l

a ck o

f

c

l

a rit_y o n the r e

l

at

i

v e _pr e s e n c e of the s e _gr a n u

l

o c_yte s

i

n

s_putu

m

of ato_{p i}c s u

b

j

e cts c o

m

pa r e

d

to a sth

m

at

i

c a n d he alth y s ub

_j

e cts Co

m

_pa r e

d

to

he alt_{h y} s u

b

_j

e cts, e o sin op

h

i l ic

i

nf la

m m

atio n

i

n sputu

m h

a s

b

e e n

w

el l -

d

o c u

m

e nte

d

in

a st

h m

at

i

c s,

b

ut r e s u

l

ts v a ry o n

d if f

e r e n c e s

i

n

i

n

f l

a

m m

at

i

o n

b

etwe e n a st

h m

a a n

d

atop

i

c

s u

b

j

e cts

Onl y a few stu

d

ie s

h

a v e c o

m

_pa r e

d

both the a

b

s o

l

ute n u

m b

e r of c e

l l

s, w

h i

ch

c o r r elate s w it

h

the

m

a_gn itu

d

e o

f i

nf la

m m

atio n, a n

d

the pe r c e ntage of c el ls,

w

h ich

ind ic ate s the r elativ e t_{y p}e s of in

fl

a

m m

atio n,

i

n sputu

m

^'^ '

H

ow e v e r, n o stu

d

y ha s

c o

m

_pa r e

d

the a

b

s olute n u

m b

e r a n

d

_pe r c e nta_ge o

f

c el

l

s a

m

o n_{g h}e a

l

t

h

_y, atop ic a n

d

a sth

m

atic c o

h

o rts at

b

a s e

l i

n e A

d d

itio n al l y, to the a utho r '

s

k

n ow

l

e

d

_ge, n o stu

d

ie s

h

a v e

c o

m

_pa r e

d i

n

i

tial s_putu

m w

e

i

_g

h

t, s ele ct sputu

m w

e

i

g

h

t a n

d

pe r c e nt c e

l l

via

b i l i

ty a

l

o ngsi

d

e

t

h

e a

b

s o

l

ute n u

m b

e r a n

d

_pe r c e nta_ge of c e

l l

s in s_putu

m

,

w

h ic

h

c o u

l d

hel p in

di

c ate the

r elativ e de_gre e of c el lular infi ltr ate in the r e s_{p i}r ato r_y tr a ct at ba s el in e T o u nde r sta nd

r e_ga r

d

in_g

d i f

fe r e n c e s

i

n airwa_{y i}n

fl

a

m m

atio n at ba s el in e a

m

o n_g r e

l

ativ e

l

_{y l}a r_ge c o

h

o rts

o

f h

e alt

h

_y, atop ic a n

d

a sth

m

atic s ub

j

e cts.

D

O b

e s

i

t

_y

an

d

I

n

f l

a

m

m

at

i

o n

T

h

e _pr e v ale n c e o

f

a sth

m

a a n

d

o

b

e sit_y

h

a s

i

n c r e a s e

d

c o n c u r r e nt

l

_{y i}n

d

e v e

l

o_pe

d

n atio n s o v e r the _pa st fe

w

de c a

d

e s^ ^

Both a sth

m

a

"_*

' ^ "'^ ^ "^ ^

a n

d

obe si_{t y}^' ^ ^' ^ ^ ■* '

a r e

c o n s

i d

e r e

d

to

b

e chr o nic _pr o-i_n

fl

_a

m m

_{ato r}

y state s c a u sin_g o xi

d

ativ e str e s s to t

h

e

b

o

d

_y.

Se v e r al stu

d

ie s ha v e show n a s s o ciat

i

o n s

b

et

w

e e n o

b

e sit_y a n

d

a st

h m

a _pr e v a

l

e n c e "

a n

d

o

b

e sit_y a nd air

w

a_{y h y p}e r r e s_po n siv e n e s s^ ^' '_*'_*

Wh

e n

f

u rthe r e vi

d

e n c e

d b

_{y p}r o s_pe ctiv e

c o

h

o rt stu

d i

e s s

h

ow

i

n_g

i

n

d i

v

i d

u a

l

s w i t

h h i

_g

h

Bo

d

_y

M

a s s In

d

e x

(

B

M

I

)

h

a v in_g a _gr e ate r

r

i

s

k

o

f d

e v e

l

o_{p i}n_g a sth

m

a

'

*^''_{* ^}

, t

h

e s e stu

d

ie s in

d

ic ate t

h

at h i g

h

B

M

I

i

s a po s si

b

le r

i

s

k

f

a cto r

f

o r a sth

m

a T

h

e r elatio n s

h

i p

b

et

w

e e n o

b

e sit_y a n

d

ato_{p y i}s le s s cle a r ^ "

, a s stu

d

ie s

ha v e fo u n

d

a n a s s o ciatio n

b

et

w

e e n o

b

e sit_y a n

d

ato_{p y} w h i le s o

m

e stu

d

ie s

h

a v e r e_porte

d

n o a s s o c

i

atio n

'_'*"'*^

Ho

w

e v e r, C i pr a n

d i

et al n ote

d

lo

w

e r B

M

I v alu e s in he alt

h

y c o ntr ols

c o

m

_pa r e

d

to ato_{p i}c a n

d

a st

h m

atic s u

b

j

e cts^

^

T he inte r_{p l}a_y

b

etwe e n o

b

e sit_y a n

d

a sth

m

a

m

a_y

b

e li n

k

e

d

with c

h

a n _ge s in

infl a

m m

atio n^ ^' '_{* ''}

B y a ctiv atin_g

w

h ite a

d

_{i p}o s e tis s u e

(W

A T

₎

, o

b

e sity is

b

el ie v e

d

to c a u s e

lo

w

-gr a

d

e s_yste

m

ic infl a

m m

atio n fr o

m

a

d

_{i p}o c_yte s

₍

fat c el ls

)

,

m

a c r op

h

age s a n

d

othe r

infl a

m m

a_to r_{y p}r o c e s s e s of the in n ate i

m m

u n e s_yste

m

that c o ul

d b

e a s s o ciate

d

w

i

th

e n

h

a n c e

d

a

i

r

w

a_y

i

n

fl

a

m m

at

i

o n °

' ^ ^ '^ ^

' ^^ "_{^ ^}

S

e v e r a

l

c r o s s

-s e ct

i

o n al stu

d

ie s

h

a v e e x a

m i

n e

d

the r elatio n s_{h i p b}et

w

e e n o

b

e sit_y a n

d

infl a

m m

atio n in s_putu

m

To

d

d et al fo u n

d

n o

a s s o ciatio n

b

et

w

e e n B

M

I a n

d

in

fl

a

m m

at

i

o n

i

n s_putu

m i

n a st

h m

atic s a n

d

n o n

-a st

h m

atic s

(

t

h

a_t

h

a

d

c

h

r o n

i

c c o u_g

h

o r c

h

r o n

i

c o

b

str u ct

i

v e _pu

l

mo n a r_y

d i

s e a s e

)

"* "

, a

l

t

h

o ug

h

t

h

e

L e s s a rd et al fo u n d n o a s s o c

i

atio n

b

et

w

e e n o

b

e sit_y a n

d

t

h

e _pe r c e nta_ge of e o sin o_p

h

i ls,

n e utr o_p

h

i ls a n

d m

a c r o_{p h}a_ge s

i

n s_putu

m

^'

,

w h il

e v a n Ve e n e tal n oted that o

b

e s e patie nts

h

a

d

a si gn

i fi

c a n tl_y

l

owe r _pe r c e nta_ge o f e o sin o_{p h i l}s a n

d

n e utr o_{p h i}

ls

i

n s_putu

m

c o

m

_pa r ed

to n o n

-o

b

e s e

i

n

di

vi

d

u a

l

s '"

Wh

e n

m

e a s u rin_g

b l

o o

d

c o u nts,

h

o

w

e v e r,

N i

e

m

e n et a

l

fo u n

d

thatB

M

I

w

as c or r elated w ith le uko c_{y t}e c o u nts ' ^

T here is a n e ed to fu rther in v e st_{i g}ate

the r e

l

at

io n s

h i

_p

b

etwe e n o

b

e sit_y a n

d i

n

fl

a

m m

atio n a n

d

a n_y

d if

f

e r e n c e s that

m

a_y e x

i

st

b

et

w

e e n

h

e a

lt

h

_y

i

n

d iv

i d

u a

l

s a n

d in

d i

v

i du a

ls

w i

th _pr e-_{e x}

i

_st

i

_n

g a

i

r

w

a_y

di

s eas e s

E

A i

m

s

T he _pri

m

a r_y ai

m

s of t

h

is stud y a r e to

(

1

)

a s s e s s the

d

i f fe r e n c e s in the a

b

s o

l

ute

n u

m b

e r a n

d

pe r c e nta_ge of n e utr o_p

h

i ls,

m

a c r op hage s, a n

d

e o s

i

n op

h i l

s at

b

a s e

l

in e a

m

o ng

h

e alth y, atop

i

c a n

d

a st

h

matic

in

di

v

i du a

ls a n

d

(

2

)

to

d

ete r

m i

n e

i f

t

h

e r e is a n a s s o c

i

at

io n

b

et

w

e e n B

M

I a n

d

the s e v a ria

b

le s at

b

a s e

l i

n e t

h

at is

m

o

d

e r ate

d b

_y

di

s e a s e statu s

T he s e c o n

d

a r_y ai

m

s o

f

th is stud y a r e to a s s e s s

di f f

e r e n c e s

i

n in itial s_putu

m

w

ei g ht, s e

le c

t sputu

m

we

i

g ht, total c e

l

l c o u nt, a n

d

pe r c e nt c el lv

i

a

b

i l

it

y at

b

a s el

in e a

m

o ng

h

e alt

h

_y, atop ic an

d

a st

h m

at

i

c

i

n

di

vi

du a

ls a n

d

to

d

ete r

m i

n e i

f

the r e

i

s a n a s s o ciatio n

M

e

t

h

o

d

s

A

.

G

e n e r a

l S

c r e e n

(

G S

)

P

r otoco

l

T he

G

e n e r al Sc r e e n

(

G

S

)

Pr oto c ol is a s c r e e n

i

n_{g p}r oto c o

l

at t

h

e Ce nte r

f

o r

En v

i

r o n

m

e ntal

M

e

d

ic

i

n e, A st

h m

a a n

d

Lu ng B

i

o

l

og y

(

CE

M

A L B

)

at t

h

e Un

i

v e r s

i

ty o

f

N

o rt

h

Ca r o

l i

n a at

C h

a_pe

l H

i l l t

h

at c ate_go rize s s u

b

j

e cts a s ato_{p i}c o r a sth

m

atic,

m

e a s u r e s

lu n _{g f}u n ctio n a n d

d

e_gr e e o

f

a

i

rwa_y

i

n

fl

a

m m

atio n, a n

d

e v a

l

u ate s the s u

b

j

e ct '

s a

b

i l it_y to

pr o

d

u c e s u

ff

i c

i

e nt s_putu

m

s a

m

_p

l

e s

f

o r a n al ysis

G

S is u s e

d

to

d

ete r

m

in e s u

b

j

e ct

el i g

i b

i l it_y

f

o r o n_goin_g a n

d f

utu r e C E

M

A L B stu

d

ie s A

l

a r_ge

d

ata

b

a s e of o v e r 5 0 0

s u

b

j

e cts t

h

at u n

d

e r

w

e nt a

G

e n e r a

l S

c r e e n

(

G S

)

at t

h

e

C

E

M

A L B

f

r o

m

M

a r c

h

2 0 0 2 to

M

a r ch 2 0 1 0

w

a s u s e

d

to

i d

e nt

i f

_{y 9 4}

h

e a

l

t_{h y}, 1 1 0 atop

i

c

(

n o n

-_{a s}t

h m

_ati_c

)

a n

d

5 3 a st

h m

atic

s ub

j

e cts fo r th is stu

d

_y Fr o

m

a tota

l

of 2 5 7 stu

d

_y s u

b

j

e cts, 1 1 7 we r e

m

a

l

e, 1 3 5

w

e r e

f

e

m

ale a n

d

5 s u

b

j

e cts ha

_{d m i}

s sin_g

d

ata T

h

e r e w e r e 1 7 3

W

h

i

te, 6 3 B

l

a c

k /

A

f

r

i

c a n

A

m

e r

i

c a n , 1 1 As

i

a n, 3 A

m

e ric a n In d ia n

/

A la ska

N

ativ e, a n

d

2

M

i

x e

d

s u

b

j

e cts a n

d

5

s u

b

j

e cts

w i

t

h m

is s

i

n_g

d

ata Bo

d

_y

M

a s s In

d

e x

(

E

M

I

)

c

l

a s s

i f

i c at

i

o n s

w

e r e

b

a s e

d

o n

U

n

d

e r

w

eig

h

t, No r

m

a

l

,

O

v e r

w

e

i

g ht, a n

d

O

b

e s e c

l

a s si

f

i c at

i

o n s o

f

t

h

e

W

o r

l d H

e a

l

t

h

O

r_ga n

i

z at

i

o n' * T

h

e r e

w

e r e 3 u n

d

e r

w

ei g

h

t, 1 4 3 n o rma

l

, 7 0 o v e rwei g ht a n

d

4 0 obe s e

s u

b

j

e cts a n

d

1 s u

b

j

e ct w

i

t

h m

is sin_g

d

ata in t

h

e stu

d

_y

S

_pe ci

f

i c

d

e

m

o_gr a_{p h}

i

c in

f

o r

m

at

i

o n

fo r e a ch d is e a s e c oho rt c a n be fo u nd

i

n T ab le A l in the A p pe n d

i

x

B

.

G S C

r

i

te r

i

a

To

b

e c o n si

d

e r e

d f

o r t

h

e

G S

_pr o c e

d

u r e, s u

b

j

e cts

h

a

d

to

b

e

b

etwe e n 1 8 a n

d

5 0

ye a r s of a_ge w ith n o r

m

al vital si gn s,

w

ith n o infe ct

i

o n s

f

o u r we e

k

s prio r to the pr o c e

d

u r e,

a n

_d

ato_{p y}

₎

S

m

oke r s

w

e r e

in c

lu

de

d in

the

G

S In

d

ivi

d

u als

w

e r e e x clu

d

e

d

i f the_y

h

ad

th_yr o

i d d is e a s e o r

h

a

d

tr e ate

d

o r s_y

m

_pto

m

atic ir ritab le

b

o

w

el s_yn

_d

r o

m

e In

di

vi

du a

ls w ith

le s s s e v e r e i

l l

n e s s e s s u c

h

a s tr e ated

de

_pr e s sio n o r h y pe rte n s

i

o n

(

w

i

tho ut

b

eta

b

lo cke r s

)

we r e in clu

de

d

Patie nts tak in_g n o n

-e x clu sio n a r_y

m

e

d i

c atio n s ha

d

to

h

a v e

b

e e n o n a sta

b le

re_{g i}

m

e fo r at le a st thr e e

m

o nths Ind iv_{i d}u als tak in_g o r al ste r oi ds fo r a sth

m

a o n e _ye ar _prior

w

e r e e x c

l

u

de

d

T ho s e ta

k in

_g

le u

kotr

i

e n e in

h

i

b

ito r s we r e a s

k

e

d

to r efr a

i

n

fr o

m

u s e 2 4

h

o u r s _pr

io

r to a

ra

et

h

a c

h

o

l i

n e c

h

a

l l

e n _ge

A H s ub

_j

e cts u nde r

w

e nt s_{p i}r o

m

etr_y to e v alu ate _pul

m

o n a ry fu n ctio n a nd s_putu

m

in

d

u ctio n S putu

m

wa s _pr o c e s s e

d

in a sta n

d

a r

di

z e

d m

a n n e r a s

de s c r

i b

e

d b

e

l

o

w

a n

d

the

m

e a s u r e s of init

ia

l a n

d

s ele ct s_putu

m w

e_{i g}

h

t, total c e

l l

c o u nt, a

b

s o

l

ute n um

b

e r s a n

d

pe r c e nta_ge o

f

n e utr o_{p h}

i l

s,

m

a c r op

h

age s, a n

d

e o sin op h i ls, a n

d

pe r c e nt c el l via

b i l

ity

w

e r e

e nte r e

d i

nto the

G S da

ta

b

a s e

D is e a s e statu s

w

a s c o nfi r

m

ed fo r all s ub

j

e cts Ato_{p i}c s ub

j

e cts te sted po sitiv e fo r a

s

k

in _pr

ic

k

te st to o n e o r

m

o r e o

f

t

h

e fo

ll

o

w

in_g c o

m m

o n al le r_ge n s :

du s

t

m

ite, c o c

k

r o a c

h

,

tr e e s, gr a s s,

w

e e

d

,

m

ol

d

, c at, guin e a p i g, r a

b b

it,

d

og, r at, a n

d m

o u s e Asth mat

i

c s u

b

j

e cts

h

a

d

a _po s

it

iv e

m

etha chol

in e c

hal le n_ge, i e P

C

2 0f ev i < ₈

_m

g

/ m

l H e alth y s u

b

j

e cts te ste

d

n e_gat

iv e

f

o r

b

ot

h

the s

k i

n _pric

k

te st a n

d

t

h

e

m

et

h

a c

ho

l

in e c

h

a

l l

e n_ge T

h

e

G S

stu

d

_y

pr oto c ol

w

a s a_{p p}r o v e

d b

_y the _O

ffi

c e of

H

v ima n R e s e a r c

h

Ethic s at t

h

e

U

n

i

v e r s

i

t_y o

f

No rt

h C

a r o

l i

n a at C ha_pe

l H

i l l

C

.

E

x c

l

u s

i

o n

C

r

i

te r

i

a

f

or

A

n a

l

y

s

i

s

Fo r the _pu r_po s e s of th is a n al ysis, s

m

oke r s

w

e r e e x clu

d

ed i

f

t

h

ey

h

a

d

te n pa ck

pr o

d

u c e a n in

_d

u c e

d

s_putu

m

s a

m

_{p l}e o r i f the s a

m

_{p l}e

h

a

d

a h i g h le v el of s al iv a r_y

c o nta

m i

n at

i

o n w it

h

n o v

i

s

i b

le s_putu

m

_{p l}u_gs Su

b

j

e cts

w

itho ut

d

ata fo r total c el

l

c o u nt a n

d

abs olute n u

m

be r s a n

d

_pe r c e nta_ge s of n e utr o_{p h i l}s,

m

a c r op hage s a n

d

e o sin op h i ls,

d

u e to

eithe r a

h i

_{g h l}e v el of s_qu a

m

o u s c el

l

c o nta

m i

n at

i

o n o r u n r e a

d

a

b l

e sl i

d

e s,

w

e r e a

l

s o

e x cluded Su_b

_j

e cts

w

e r e in clu_de

d

a s a sth

m

atic s

i

f the_y

d

i d n ot ha v e n o r

m

al lu n_{g f}u n ctio n

at

b

a s el in e a n

d

the r e

f

o r e

d

i

d

n ot u n

d

e r_go a

m

et

h

a chol in e cha

l l

e n_ge,

b

ut ha

d

a

h i

sto ry of

a st

h m

a

D

.

B

o

d

y

M

a s s

I

n

d

e x

(

B

M

I

)

B

M

I

f

o r e a c

h

s u

b

j

e ct wa s c alc u

l

ate

d b

_y

d

ivi

d

in_g the s u

b

j

e ct

'

s

w

e_{i g h}t in k i lo_gr a

m

s

b

_y

h

ei g

h

t in

m

ete r s s_qu a r e

d

u s

i

n_g o

b

j

e ctiv e he

i

_{g h}t a n

d w

e

i

_g

h

t

m

e a s u r e

m

e nts Ba s e

d

o n

the Inte r n atio n al C la s si fi c atio n _gui del in e s of the

W

o rl

d H

e alth Or_ga niz atio n , B

M

I v alu e s

w

e r e cla s si fi e

d

into t

h

e fol low in_g c ate_go rie s :

U

n

d

e r

w

e

i

_g

h

t

₍

B

M

I < 1 8 5

₎

;

N

o r

m

al

(

1 8 5 0

- _{2 4 9 9}

)

; Ov e r

w

e_{i g h}t

₍

2 5 0 0 - _{2 9 9 9}

)

; O

b

e s e

(

> 3 0.0 0

)

'

^

E

.

G S P

r o c e

d

u r e

f

o r

I

n

d

u c e

d S

p

utu

m

P

r o c e s s

in

g

In

d

u c e

d

s_putu

m

s a

m

_{p l}e s

w

e r e obtain e

d

u sin_g a n ultr a s o nic n e

b

ul ize r to

a

d m

in

i

ste r 3

%

s al in e fo r 7

m i

n ute s

f

ol lo

w

e

d b

_y a c

l

e a n s

i

n_{g p}r o c e

d

u r e i n

w h i

ch s ub

_j

e cts

ri

n s e

d

t

h

e

i

r

m

o uth, ga rg le

d

, s c r ape

d

a n d

b

lew t

h

eir n o s e Su

b

j

e cts

w

e r e a s

k

e

d

to c o ug h

s_putu

m

into a c u_p

U

_po n a n o r

m

al Pul

m

o n a r_{y F}u n ctio n Te st

₍

P F T

₎

, t

h

e pr o c e s s

w

a s

r e_pe ate

d

tw

i

c e

w i

t

h

4

%

a n

d

5

%

s a

l i

n e, r e spe ct

i

v e

l

y Ast

h m

at

i

c s u

b

j

e cts

w

e r e tr e ate

d

w

i

t

h

al

b

ute r o

l

to a v oi

d

br o n cho s_pa s

m

s _p

ri

o r to t

h

e s_putu

m

in

d

u ct

i

o n _pr o c e

d

u r e

T he

i

nit

i

a

l

s_putu

m w

e

i

_g

h

t

₍

m

_g

₎

w

a s r e c o r

d

e

_d

_{p ri} o r to s ele ctin_g s_putu

m

_{p l}u_gs w

i

th

0 1%

d

ith iotr e

i

to

l

(

D T T

) (

m

L

)

e_qu al to fo u r ti

m

e s t

h

e

w

ei g

h

t

₍

m

_g

₎

of the s ele ct s a

m

_{p l}e

T

h

e

m

ixtu r e

w

a s tu

m b

le

d

fo r fift e e n

m

in ute s at r o o

m

te

m

_pe r atu r e, d i luted w ith I X

Du

l b

e c c o '

s P ho s_{p h}ate

-Buf

f

e r e

d

Sal in e

₍

D P B S

_{) (}

m

L

₎

e_qu a

l

to the v o

lu

m

e o

f

D T T

pr e vio u s_ly a

d de

d

, tu

m b

le

d f

o r

fi

v e

m

in ute s at r o o

m

te

m

pe r atu r e, a n

d

pa s s ed t

h

r o ug

h

a 5 3

n

m

n_ylo n

m

esh fi lter

(

S

m

al l Parts, In c

)

T he

m

ixture

w

a s c e ntri fuged at 5 0 0 X g fo r 1 0

m

in ute s a n

d

the c e

l l

_pel

l

et

w

a s r e-_{s u s}

pe n

d

e

d

in

H

a nk'

s Bu

f

fe r e

d Sa

lt Solut

i

o n

₍

H

B S S

₎

T he total c el l c o u nt a n

d

_pe r c e nt c el l via

b

i l it_{y w} e r e e v alu ate

d b

_y a te chn

i

cia n u s

i

n_g

a he

m

o c_{y t}o

m

ete r Fo u r sl i

de s

w

e r e c r e ated u sin_g a 6 0 |j L c el l s u s_pe n sio n at a

c o n c e n tr at

i

o n o

f

1x 1 0 c el ls

/m

L in a s

i

n _{g l}e c_{y t}ofu n n el a n

d

stain e

d w

ith a

H

e

m

a 3

sta

in e

d

s et

f

o r 5 0 0 tota

l

c el

l

c o u nt A r e s e a r c

h

s_pe cia

l i

st e v alu ate

d

the c e

l

l

d

i

ff

e r e ntia

ls

u sin_g

m i

c r o s c o_{p y T h}e a

_b

s olute n u

m b

e r s of n e utr o_p

h

i ls,

m

a c r op hage s a nd e o sin op

h

i ls

w

e r e e x_pr e s s e

d

a s t

h

e tota

l

n u

m b

e r of c e

l l

s _pe r

m i l l i

_gr a

m

a n

d

t

h

e _pe r c e nta_ge o

f

n e utr o_p

h

i

l

s,

m

a c r op

h

age s a n

d

e o s

i

n op h

i l

s

w

e r e e xpr e s s e

d

a spe r c e ntage s o

f

n o n

-s_qu a

m

o u s c el ls^ Cel l c o u nts we r e e v alu ate

d b

_y a te c

h

nic

i

a n

w

ho

w

a s

b

l

in

d

to t

h

e

i d

e nt

i

t_y

a n

d d

is e a s e statu s of the s ub

j

e ct a n

d

o nl y

i d

e nti fi e

d

the s u

b

j

e ct

b

_y a _pr e vio u sl y a s s

i

_gn e

d

n u

m b

e r

F

.

S

tat

i

s

t

i

c a

l A

n a

l

y

s

i

s

T

h

e _pri

m

a r_y s_putum v a ria

b

le s w e r e the a

bs o

lute n u

m

be r s a nd pe r c e nta_ge s of

n e utr o_p

h i l

s,

m

a c r op

h

age s a n

d

e o s

i

n op h i ls T he s e c o n

d

a ry sputu

m

v a ria

b

le s

w

e r e total

c el

l

c o u n t,

in

i

t

i

al sputu

m w

ei g

h

t, s e

l

e ct sputu

m w

ei g

h

t a n

d

pe r c e nt c el l v

ia

b

i l ity N atu r a

l

lo_g tr a n sfor

m

atio n s of bot_{h p}ri

m

a r_y a nd s e c o nda r_y s_putu

m

v a riab le s

w

e r e u s ed i n th is

y

)

wa s _pe r

f

o r

m

e

d

, w he r e y

w

a s equ a

l

to a spe ci fi c sputu

m

v a ria

bl

e A l l log

tr a n sfo r

m

atio n s r efe r e n c e

d

in this _pa_pe r r e

f

e r to the lo_g tr a n sfo r

m

atio n of

₍

1+ _y

₎

O

n e-

w

_a

y An a

l

_y sis o

f

Va ria n c e

(

A N O V A

)

w

a s _pe rfo r

m

e

d

to c o

m

_pa r e a_ge, B

M

I

a n

d

a

l l

s_putu

m

v a r

i

a

b l

e s a

m

o n_g

h

e a

l

th_y, atop

i

c a n

d

a st

h m

atic c o

h

o rts In

d i

v

i d

u al

d i f fere n c e s in

m

e a n s of a_ge, B

M

I a n

d

sputu

m

v a ria

b

le s a

m

o ng two spe ci fi c c oho rts

w

e r e

a s s e s s e

d

u s

i

n_g a _pa r ametr

i

c two-t_a

i l

_e

d

/-_{te st}

To e v alu ate

h

o

w

B

M

I af

f

e cte

d

the s e

d

i

f f

e r e n c e s,

m

ultiv a r iate l in e a r r egr e s sio n

a n a

l

_ys

i

s

w

a s _pe rfo r

m

e

d

to

f i

n

d

the e

f f

e ct o

f

B

M

I o n s_putu

m

v a ria

b

le s a s

m

o

d

e r ate

d b

_y

di

s e a s e statu s, ad

j

u sting fo r the c o ntr ol v a ria

b

le s age, s e x a n

d

r a c e Both

m

ale s a n

d

W

h it

e s s e r v ed a s the r efe r e n c e _gr o u_ps

f

o r s e x a n

d

r a c e

T he

m

ult_{i p}

l

e l in e a r r e_gr e s sio n

m

o

d

el

w

a s :

yi = a +

P

xB

M

I +

l

^zAtop ic +

^

i Ast

h

m

a tic +

p

^_iB

M

l * Atop ic +

p

_^B

M

l *

Ast

h m

atic +

_P

_{e^ g}e +

p

-^Sex +

P^

A

m

e r ic a n ln d ia n

/

A la skaNa tiv e +

p

,₎Asian +

P

^ oB la ck

/

A

f

r ic anA

m

e r ic an +

p^

M

ix e

d

+ Si, w he r e yj

w

as a sputu

m

v a r

i

a

b l

e

P

4 a n

d

P

s

w

e r e the inte r a ctio n te r

m

s

b

et

w

e e n B

M

I a n

d

the atop

i

c c oho rt an

d

b

et

w

e e n B

M

I a n

d

t

h

e a sth

m

atic c o

h

o rt, r e s_pe ct

i

v e

l

_y Bot

h

the e

f f

e ct o

f

B

M

I o n s_putum

v a ria

b

le s a nd the e

ff

e ct of

di

s e a s e statu s o n s_putu

m

v a r

i

a

b

le s fo r

d

i

f

fe r e nt le v els o

f

B

M

I

w

e r e a n al yz e

d

u sin_g the

f

ol low

i

n_g e_qu atio n s A ge, s e x a n

d

r a c e

w

e r e r efe r r e

d

to a s +

1

E

_ff

e ct o

f

B

M

I

o n

Sp

utu

m V

a ria

b l

e s

Ca s e 1: Ato_{p i}c = 0 a n

d

A sth matic = 0

(

H

e alth y

Su

b

_j

e cts

)

yi= a +

f

i

-i_B

M

l +

^

2

(

0

₎

+

_^

₃

₍

0

₎

+

_P

^B

M

I

i

O

)

+

p

^B

M

I

i

O

)

+ c on tr ols + £ (

yj_ a +

P

iB

M

I + c on tr ols + Si

In he a

lt

h y s u

b

j

e cts, the ef fe ct of B

M

I o n

[

sputu

m

v ari ab le

]

is

Pi

C

a s e 2: Ato_p

i

c = 1a n

d

Asth mat

i

c = 0

(

Ato_p

i

c

Su

b

j

e cts

)

3/₁₌ a +

_f

i

yB

M

l +

/

?2

(

1

)

+ jSg

C

O

)

+

P

4B

M

K

I

)

+

p

sB

M

I

(

0

)

+ c on tr ols + S i

yi= a +

p

^B

M

l +

^

2 +

P^

B

M

I + c on tr ols + e ^

yi^

(

a +

/

?2

)

+

(

/

?i +

p

4

)

B

M

I + + c on tr ols + e ^

In ato_{p i}c s u

b

j

e cts, t

h

e e

ff

e ct o

f

B

M

I o n

[

sputu

m

v a

ri

a

b l

e

]

is

( P

i +

P

4

)

.

Ca s e 3: Ato_{p i}c = 0 a n

d

A sth matic = 1

(

A st

h m

atic Su

b

j

e cts

)

yi= a +

p

-^B

M

l +

^

2

(

0

)

+

/

?3

(

1

)

+

/

?45

M

/

(

0

)

+

j

i^B

M

l

iX

)

+ c on tr ols + e ^

yi= a +

p

^B

M

l +

^

3 +

p

^B

M

l + c on tr ols + £ j

y i^

(

a +

/

?3

)

+

(^

1 +

/

?5

)

B

M

I + c on tr ols + £ ;

2 E

_ff

e ct o

f

D

is eas e

St a t

u s o n

Sp

utu

m Va

r ia

b l

e s

f

o r

D

i

ff

e r e nt

L

e v e

l

s o

f

B

M

I

Ca s e 1:E f fe ct of

b

e

i

n_g ato_{p i}c r elativ e to he alth_y o n s_putu

m

v a

ri

a

b

le

C ha n _ge in

_[

s_putu

m

v ari a

b

le

]

fo r a 1

-u nit in c r e a s e in Ato_p

i

c =

[

a +

_P

_^B

M

l +

_P

₂

_O

-

)

+

P

^Ast

h

m

a tic +

P

^B

M

I

(

1

)

+

f

^

sB

M

l * Ast

h m

a tic + c on tr o

l

s + f

J

-

[

a +

P

xB

M

l +

P

2

W

+

P

^Asth

m

a tic +

P

^B

M

I

(

Q

)

+

P

s B

M

I * Asth

m

atic + c on tr ols +

£ i

h [P

2 +

M

M

I

]

T

h

e e

f f

e ct o

f b

e

i

n_g ato_p

i

c r e

l

ativ e to

h

e alt_{h y} o n

_[

sputu

m

v a

ri

a

b l

e

]

is

( P

₂ +

_P

₄ B M I

₎

C

a s e 2:E

f f

e ct o

f b

e

i

n_g a sth

m

atic r e

l

ativ e to

h

e a

lt

h

y o n a s_putu

m

v a r

i

a

b l

e

C

h

a n _ge in

_[

s_putu

m

v ari a

b l

e

]

fo r a 1-_{u n}

i

t

i

_{n c r e a s e}

i

_n A_sth

m

_ati_{c =}

=

_[

_{a +}

/

?_i 5

M

/ +

_p

_^Ato_{p i}c +

_/

?₃

(

1

)

+

_p^

B

M

I * _Ato_{p i}c +

p

^B

M

I

i

l

)

+ c on tr ols +

f

j

]

-

[

a +

p

^B

M

I +

p

2^ top ic +

/

?

3

(

0

)

+

p

^B I^I I * Atop ic +

p

sB

M

I *

(

0

)

+

c on tr ols + £_i

]

=

[

_/

?

₃ +

I

^

^B

M

I

]

T

h

e e

f

fe ct o

f b

ein_g a st

h m

atic r e

l

ativ e to he alt

h

_y o n

[

s_putu

m

v ari a

b

le

]

i

s

_{( P}

3 +

P

5B M I

)

3 E

_ff

e ct o

f

A g

e,

S

e x an

d R

a c e

C

o n tr o

l

s o n

Sp

u tu

m Va

r ia

b l

e s

A ge : T

h

e e

ff

e c to

f

a_ge o n

[

s_p utum v a r iab

l

e

]

is

/

J_g

S

e x : T

h

e e

ff

e ct o

f

b

ein_g

f

emale r e

l

a tiv e to male o n

[

s_p utum v a r ia

b

le

]

is

^

_j

Ra c e :

T

h

e e

ff

e cto

f

b

ein_{g A m}e ric a n In

d

ia n /

A

la s

k

a

N

ativ e r elativ e to

Wh i

te o n

[

s_putum

v a r iab

l

e

_]

is

_^

_g

T

h

e e

ff

e ct o

f

b

ein_g

A

sia n r elativ e to

W h

ite o n

[

s_p utum v a ria b le

]

is

f

i

_g

T he e

_{f f}

e ct o

_f

bein_{g B}

l

a c

k

/ A

f

ric a n A me ric a n r e

l

a tiv e to

W

h ite o n

[

s_putum va riab le

]

is

_/

? i o

4 A

c c o u n t in

_g

f

o r

L

o

_{g T}

r an s

f

o r

m

at io n s

To c a

l

c ulate _pe r c e nt cha n_ge in a s_putu

m

v a

ri

a

b

le

f

o r a 1

k

_g

/ m

'

i

n c r e a s e in B

M

I in

the r e_gr e s sio n a n a_{l y}sis, the

f

ol lo

w i

ng c a

l

c u

l

atio n s

w

e r e pe r

f

o r

m

e

d

:

Fo r a 1

k

_g

/m

^

in c r e a s e in B

M

I, t

h

e sputu

m

v a ria

b

le in c r e a s e

d

fr o

m l

og

(

y + 1

)

to

lo_g

(

_{y +} 1

)

+

p

T hu s, log

(

y + 1

)

+

P

= _lo

g

(

y + 1

₎

+ lo_g

(

e'^

)

= lo_g

₍₍

_{y +} l

)

*e''

₎

. T he r efore

,

t

h

e _pe r c e nt c

h

a n_ge

f

r o

m l

o_g

(

_y+ l

₎

to

l

o_g

₍

_y+ l

₎

+

_p

w

a s :

/

₍

e P_(y+l₎- ₍

y+l₎

)

N

, „ ,

Pe r c e nt c

h

a n_ge in s_putu

m

v ari a

b l

e = 1 0 0 *

I

^^

—

1

= _{1 0 0} *

(

e' * — 1

)

T hu s, a I

k

g

/m

^

i

n c r e a s e

i

n B

M

I is a s s o c

i

ate

d w

it

h

a 1 0 0*

(

e^- 1

)

_pe r c e nt c

h

a n_ge

in a s_putu

m

v a

ri

a

b

le Be c a u s e t

h

e

l

o_g tr a n s

f

o r

m

atio n

w

a s

(

_y + 1

)

, th is pe r c e nt c

h

a nge

a ctu al l y r e

fl

e cts a c

h

a n_ge

i

n

(

_y + 1

₎

,

W

i e r e y

w

a s a sputu

m

v a

ri

a

b l

e T

h

u s, a

l l

pe r c e nt

cha n_ge s r e

f

e r e n c e

d

in _t

_{h i}

s _pa_pe r r e

f

e r to a cha n_ge in

(

_y + 1

)

, ^ \iae r e y

w

a s a sputu

m

v ari a

b

le

A N O V A

w

a s _pe rfo r

m

e

d

u s

i

n_g

S

A

S

v9 2

(

S

A S In c,

C

a ry, N

C

)

Regr e s sio n

a n al ysis

w

a s _pe rfo r

m

e

d

u sin_{g S A S}v9 2

(

S A S In c, Ga ry,

N

C

)

a n

d

S T A T A I O

(

StataCo rp

L P,

C

o

ll

ege

S

tat

i

o n, T X

)

Ba r gr ap

h

s

w

e r e c r e ate

d

u sing

M

i

c r o s o

ft

Ex c el

(M

ic r o s o

ft

C

o r_p, Re

d m

o n

d

,

W

A

)

a n

d

a

l

l r egr e s s

i

o n a n al ysis gr ap hs we r e c r e ate

d

u sin_{g S T A T A I O}

(

Stata

C

o r_{p L P}, Co

l

lege Statio n , T X

)

Statistic al si gni

fi

c a n c e

w

a s a c c epte

d

at the p < _{0 0 5}