18 cN8Fn4gq
Jurkat cells cultured in calcium-free RPMI-1640 medium (Gibco BRL; number O'mcN*
22300-107) containing calcium-free 10% FBS were triggered by anti-Fas IgM. The E)gD"^rex
treated cells were harvested at the indicated time points and incubated with Fluo-3-AM at q!eE~O;A
a final concentration of 1 micromolar for 30 min at 37°C (Scoltock et al., 2000). The AcPLJ!y
labeled cells (50,000 cells per treatment) were then analyzed by exciting the cells at 488 -HG.GA
nm and examining the fluorescence emission of Fluo 3 at 530 nm with a FACS Scan, . : Wf>:
(Becton Dickinson). A one micromolar concentration of LPA was used as a positive ehr-o7](
control for Ca2+ induction. The data thus obtained was analyzed with the software Win *;Mi/^pzK
MDI 2.8 and represented as contour plots.The effect of chelating intracellular calcium on t?4H9~iH
translocation of annexin I was studied by culturing Jurkat cells in the presence of 10 bOFzq
>k_
micromolar BAPTA-AM, with or without the addition of anti-Fas IgM. Cells were
D}98ZKi
harvested and fractionated as detailed above, and the S-100 fractions were assessed by K)7T]z`
immunoblotting for presence or absence of annexin I. 10Ik_L='
Mouse bone marrow transduction and transplantation. _SAM8!q4,
Retrovirus-mediated transduction of mouse bone marrow cells was done by published
usB*Wn8
methods49. Prestimulated low-density bone marrow cells were infected with high-titer >v.fH6P,}
retrovirus supernatant on fibronectin-coated plates. Retrovirus supernatant was generated CO='[1"_5
in the phoenix-gp cells with a mouse stem cell virus–based retroviral vector coexpressing
vW1^
EGFP and HA–RhoH as described50. EGFP+ sorted cells were transplanted by 7__[=)(b2X
intravenous injection into the sublethally irradiated (300 rads with a 137Cs irradiator) -3{Q`@F
Rag2-/- recipient mice. At 9 weeks after transplantation, thymus, peripheral blood, bone <l{oE?N
marrow, spleen and lymph nodes from each recipient mouse were collected for analysis [a2]_]E%
of EGFP+ chimerism and hematopoietic lineage by flow cytometry. Expression of ]#)(D-
i
HA–RhoH and HA–RhoHF73F83 in EGFP+ sorted thymocytes of recipient mice was q? '4&
confirmed by immunoblot analysis. eU1F7LS
Determination of renal morphology 9Da{|FyrD
Kidney slices were postfixed in buffered 2% OsO4, dehydrated, and embedded in an MA`nFkVK
Araldite-EM bed 812 mixture. Large sections were cut perpendicular to the renal capsule, .~f )4'T 9
containing cortex, and medulla. Thin (1 m) sections were analyzed in a blinded manner
'"B
for morphologic alterations, as previously detailed d4S4
e
Patient population pl`4&y%Me
Patients included in the study met the following criteria: (1) biopsy-proven IMN; (2) "lh4Vg\7n
creatinine clearance 30 ml per min per 1.73 m2; and (3) persistent proteinuria >5 g per tO M$'0u
24 h despite treatment with an HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A) {N4 'g_
reductase inhibitor, an ACEi, and/or ARB at maximal tolerated dose for at least 4 months. 3K
&637
The Mayo Institutional Review Board and the Research Ethical Board, University Health z9aR/:W}
Network, University of Toronto approved the study protocol. All patients gave written r%U6,7d=)
informed consent. Patients who had been on treatment with prednisone, cyclosporine, or rmX*s}B
19 $6rm;UH
mycophenolic mofetil within the last 4 months or alkylating agents within the last 6 .4zzPD
$1
months were not included in the study. Patients with active infection, diabetes, or a >v+1v
secondary cause of MN (for example, hepatitis B, systemic lupus erythematosus (SLE), \l GD8@,x
medications, malignancies) were also excluded. `MI\/oM@
Treatment Cn~VJ,l
g
At enrollment, a low-sodium (<4 g day-1) and low-protein (0.8 g per kg per day of c
MY}Y
[2c
high-quality protein) diet was recommended and patients were encouraged to maintain ;'S,JGpvT
the same diet throughout the duration of the study. All patients received a similar YV.' L
conservative treatment regimen that included loop diuretics to control edema, an dc)%5fV\
HMG-CoA reductase inhibitor, and an ACEi combined with an ARB if tolerated. Cbff:IP
-Blockers and non-dihydropyridine calcium channel blockers, in that order, were added z!g$#hmL>
when required to control systolic blood pressures to <135 mm Hg in >75% of the hEZvi
readings. Patients who after a minimum of 4 months of conservative therapy and X:i
?gRy"
maximized Ang II blockade had proteinuria >5 g per 24 h received two i.v. infusions of jFN0xGZ
rituximab at a dose of 1000 mg on days 1 and 15. To minimize infusion reactions, ##jJaSxG
patients were premedicated with acetaminophen (1000 mg) and diphenhydramine ,Cr%2Wg-
hydrochloride (50 mg) orally. In addition, methylprednisolone (100 mg, i.v.) was given %~Yo{4mHs
prior to the first rituximab infusion. B-cell depletion was defined as CD19+ count v9f+ {Y%-
<5 cells per l at any time and B-cell recovery was defined as CD19+ cell count o +$v0vg%T
>15 cells per l. Patients treated with rituximab, who at month 6 had proteinuria >3 g per b}qfOgd5
24 h and in whom CD19+ B-cell counts had increased to >15 cells per l, received a y0xBNhev
second course of rituximab treatment following the same protocol described above. ~5P9^`KNH
Follow-up v`|]57?A
In all patients, clinical and laboratory parameters including complete blood counts, cEL:5*cAU}
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum Mq0MtC6-
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry KU8,8:yY
and at months 1, 3, 6, 9, and 12. Creatinine clearance and protein and creatinine excretion 0F)v9EK(W4
in the urine were assessed by performing two consecutive 24-h urine collections at each |=q~X}DA
time point. Data were considered accurate when urinary creatinine excretion was HyzSHI
consistent with a complete 24 h collection. The mean of the two measurements was T1Py6Q,-
considered for the analysis. The presence of HACAs was evaluated at baseline and at ("a@V8M`$F
months 3, 6, 9, and 12. yY#h1
Method / Approach / Study/ Technique }d%CZnY&7
A discussion is presented of a problem-solving system ](JrEg$K
To improve the efficiency of the method, the following approach may be applied. f/Y&)#g>k
In order to an investigation was made to find the causes of the i
f&bp ,
Although large collections of rules and equations have been complied, none are generally %IO*(5f
accepted mu\1hKq;B
20 9dNB_
This approach will be explained and discussed thoroughly in the body of the report. t'2A)S
This can be accomplished by &PJ;B)b
This algorithm to compute the total cost can be described step by step as follows: $`lWW6>P
The above preliminary analysis has provided important information uw3vYYFX
Various methods have been proposed for selecting an optimum...
}ktIG|GC
These concepts have been applied to ho=!Yy
On the basis of the concept mentioned above, =2$(
tXL
This can be achieved by uX-^9t
In addition, tissues were stained for infiltrating lymphocytes (CD20 and CD3), and the 9n(68|^$
amount of interstitial fibrosis was quantified by histomorphometry. Formalin-fixed, ?>;b,^4
paraffin-embedded sections were cut onto coated glass slides. Following heat-induced O[d#-0s
antigen retrieval, sections were incubated at 20°C overnight with either anti-CD20 zTc;-,
primary antibody or anti-CD3 primary antibody, both at 1:1000 dilution (Dako, Canada 4$iS@o|
Inc, Mississauga, Ontario, Canada). After rinsing all sections, pretreatment with 3% g>t1rZ
hydrogen peroxide was performed to prevent endogenous peroxidase activation. Sections #Z
J 1\Ov
were incubated with a secondary rabbit anti-mouse antibody linked with avidin–biotin l73%
y
complex. Sections were counterstained with hematoxylin and examined by light L(|K{vH h]
microscopy. gnH{
_
The HACA assay is a proprietary bridging enzyme-linked immunosorbent assay U5C]zswL
performed at Genentech Inc. that measures the antibody response to rituximab in human !L.R"8!
serum samples. j4;^5
Dy^
In all patients, clinical and laboratory parameters including complete blood counts, b>ZAkz)U+
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum ;i[JCNiS\
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry t]SB.ja
and at months 1, 3, 6, 9, and 12. |fKT@2(
This fact suggests that a new concept m]V#fRC
This was accomplished by taking ... +IS6l*_y>6
The preparatory stage is very time consuming process. [\3ZMH
*
Test are performed for validity, completeness, and compatibility }Hb0@
b_
There is little hope of achieving successful ... gM&O dT+i
There has been an increasing awareness of the potential of using most ..so far made have IoLP*D
not taken this approach, with the exception of J.R])
&CB
Only a few studies can be found. pnuo;r s
It is a very tedious process to go through [;{xiW4V]
It is only when .. has been completed that .. may be effected )Wle
CS_
The entire interpretation process is conducted in one's head. VHPqEaR
These approaches are sometimes very tedious. kBqgz|jE%
Several techniques can be used ' ds2\gN
A polynomial parametric model can be written as [the following]/[follows]: 3ibQbk
A xx model is constructed/formulated using xx. :
jkO
A xx model represents an xx by its xx. Pl=)eq YY
A process decision model captures the logic essential to +F q_w
From the equation above, xx is equal to the summation of xx times the ... ,y'6vW`%g9
21 sSfP.R
The validity of a xx model can be checked using Euler's formula. D7nK"]HG;l
Given a model, one can mathematically determine whether ... or ... 8Tg1 >q<
Equations for xx need to be derived and implemented in the system. L{VnsY V
A number of heuristic rules have been developed for OLIMgc(W
Optimum .. techniques can be made more reliable by ... so that :[+8(~| za
An algorithm based on the characteristic ... is used to determine c
$r"q :\
Euler's formula states the following: %0 {_b68x
The completed model should agree with the formula. Y=|p}>.}
For manufacturing purposes, a detailed and precise model of the object is necessary bD=H$)
Engineering design models are very well defined; therefore, L754odc
To keep the domain narrow enough to be implementable, yet wide enough to be useful. wbJBGT{sm
Point of View &lU\9
from an implementation standpoint, %\-E
R!b
From the point of view of this application, kJl^,q
From this point of view, Zadeh suggested an inference rule named xxx (CRI for short). "P?O1
Information is the meaningful interpretation and correlation of some aggregation of data qE2VUEv5Y
in order to allow one to make decisions.
PX/Y?DP
From a practical point of view, the computational aspects of an FLC require a L7{}`O/g7
simplification of the fuzzy control algorithm. &xB*Shp,B
The use of a hammer to insert screws, although partly effective, tends to distort, destroy, a9;KS>~bq
and generally defeat the purpose of using a screw [Kusiak AI Implications for CIM yYGs]+
p.129] vu@.;-2E%
Statistical analysis @@^iN~uf
Data were analyzed by one-way analysis of variance comparing the three conditions B<L7`xL
(sham operation, ischemic AKI, and bilateral nephrectomy) at each time point. If ~n@rX=Y)]0
significant F-statistic from analysis of variance existed, this test was followed by Dunnett smfI+Z S"
post hoc multiple comparison procedure with sham operation as the control group. For all q=->) &D%
other comparisons, Student's t-test was used. A P-value of <0.05 was considered as <A=1]'1\r
statistically significant. @Q,Q"c2
Values are expressed as means s.e.m. and significance was evaluated by Mann–Whitney ~IW{^u
U test using GraphPad Prism, version 4.0 software (GraphPad Software Inc., San Diego, ^?H|RAp
CA, USA). dq.U#Rhrx
All values are expressed as means s.d. Statistical significance (defined as P<0.05) was ]w>o=<?b
evaluated using analysis of variance and Bonferroni t-tests, and the two-tailed Pearson's `dcz9 *
test, where appropriate. o$</At
Data are expressed as mean s.d., median and interquartile range, or frequencies, as JMt*GFd
appropriate. Variables who deviated from the normal distribution (positively skewed) @I-gs(
were log-transformed (log10) before the correlation study. DFWO5Y_
Data are represented as means s.e.m. Student's t-test and multiple comparisons with t-test ;$&\:-6A#
post hoc analysis of variance were used as indicated below, for the comparison of 5z0VMt
22 /a^
R$RHl'
morphological, immunohistological and functional parameters. Statistical significance }~+,x#
was set at P<0.05. xrvM}Il
The primary efficacy parameter was defined as change in urinary protein excretion from R`c[?U
baseline (week 0) to 12 months after treatment. The 12-month changes were tested D {
Ol8:
against zero using the paired t-test. Secondary end points included 6-month changes in 2H4vK]]Nl
protein; the number with PR or CR at 6 or 12 months; and changes in glomerular \t=0rFV)t
filtration rate (GFR), serum albumin, and lipid profiles. Study sample size was based on 3<W%z]k@M
the desire to have 80% power to detect a drop in urinary protein of at least 2.0 g day-1. y8s!sO
Assuming a two-sided hypothesis test performed at a significance level of 0.05 and an s.d. YS^!'IyG/B
of urinary protein change of 2.5 g, it was determined that 15 patients were required.2 ? nW>'z
Definition of remission status is according to the criteria established by Cattran et al.30 9r]|P}yuS
CR was defined as proteinuria <0.3 g per 24 h, PR as proteinuria 3 g per 24 h, and a SdYf^@%}F
>50% reduction in peak proteinuria and non-response as <50% reduction in peak
Qh&Qsyo%
proteinuria. Any patient reaching a CR or PR was considered a treatment success ?p@J7{a
The statistical significance of differences for the mean values of cytokine concentration P!|Z%H
and T cell proliferation was determined with Student's t-test. Differences with a P value @p*)^D6E\
of less than 0.05 were considered significant. RX>P-vp
Statistical significance was determined by Student's t-test. Data with a P value of less @5nFa~*K%
than 0.05 were considered significant. KCTX2eNN&h
In vitro and in vivo experiments were analyzed by the two-tailed Student's t-test, with P JnsJ]_<
values less than 0.05 considered statistically significant. O<cP1TF
The significance of differences among groups was determined by Student's t-test and Ldj
z-
one-way analysis of variance (SigmaStat software; Jandel). 2g{tzR_j
Comparisons C*(
As well, the pros and cons of these representations from a process planning point of view y,nmPX?]n
will be discussed. +>q#eUS)
The method of using xx to implement xx described by Zadeh (1973) appeared more D|9+:Y
suitable qazA,|L!
As discussed [in the previous section]/[preciously], RHC
ZP
Relation ;yJ:W8U]+;
We can not invert F' directly because it defines a many-to-one mapping. d5m-f/
The relationships appear very complicate bGWfMu=n
Lifting tasks involve complex and imprecise relationship between the task variables and >-A@6Qe_
the human operator's characteristics.
p +i1sY
These methods are based on the relationship between ... and ... |GnTRahV.
The fundamental concept of a fuzzy rating language is that we can establish a relationship YqX$a~
among terms such as high, medium, and low, and then modify these relationships. d<a|dwAeh
This article will thus mention the latter as well as the former. HK!ecQ^+
The former two bear a close relation to a fuzzy Cartesian product. )]1hN;Nz
Importance f4f2xe7\Q
) 5Kg'&B (
23
T^}UE<
The emphasis is on an implementation of a general approach to rule based decision @z@%vr=vX
making. v=N?(6T
Consideration / Attention Uq
.6h
Careful evaluation is necessary to ensure 7QsD"rL
Such a formulation does not change further considerations. NHGTV$T`1
Considerable attention has been paid to q\0/6tl_
Attention should be paid to an important finding of this investigation. $`F9e5}G
Caution should be exercised in this process to avoid ... K!-OUm5A
Primary consideration is given to ... components, though others can be accommodated
e,U:H~+]
After ... has been defined by ..., a carefully analysis is carried out/performed to determine EA& 3rI>U)
A number of factors such as ...need to be taken into consideration before making the _VrY7Mz:r
appropriate decision. iEvQ4S6tD
It should be noted that ,>
zEG
It is important to point out that ... 3'4+3Xo
These considerations have heightened interest in the possibility of providing ... )%7P?^>
We should stress the fundamental importance of the xx
qyH-Z@
Results. {OU|'
Advantages / Disadvantage hS]w
A"\87
One of the major advantages of this new measure of xx is that it can be applied to the B
71/nt9
experimental study of _wq?Pa<)e
One advantage of using a .. is the ease of preparing it. &|hK79D
It has a very fast decision making process P-@MLIC{
All the algorithms involve mostly logical operations. [xMa^A>p
It can be easily and without additional cost implemented in a microprocessor;based $T'!??|IF
environment. QU).q65p
It can reduce the waste of designing from scratch. EApKN@<"
The advantages of using a xx to represent xx are the following: (`tRJWbdz
However, xx is not without its shortcomings. /
A=w`[<
In most cases, the xxx shows an improvement over the existing xxx. Y=Vbs x
Compared to the existing xx, the impacts of the xx are generally reduced by 5% to 9%. >Fel) a
The "best case" results shows a savings of 6% to 9%. %$!3Pbui
Most of the existing works based on xx approach can only recognize a xx . )Z}AhX
Most of the above methods are computational expansive and limited to xx. ~4~`bT9
Some other advantages of xx are the following: ~Z\:Nx
The problem is the limitation of this method to a limited domain of parts. f[ ^f/jGm
It proved limited in application because it demanded precision in system modeling that %Sr+D{B
was impossible in practice. !gsvF\XDM
There are advantages to be gained in the structuring of costs and benefits, the use of xx, %Tm*^
The disadvantages of this method are also disadvantages of conventional xx approaches. ;XKe$fsa~?
This combines the best features of both techniques o5 L ^
24 0'QWa{dS\
Hopefully, this tool can be as the reference framework of for developing a xx platform, iGN\ >m}
and helping the administration, marketing, and knowledge management activities in sibYJK Oy
virtual communities. %,$ n^{v
Results dAo;y.3
An improvement on the result shown above can be made by based on the data provided .HqFdsm
Recent work has identified {@Mr7*u
Time-dependent changes of wQkM:=t5
Cyclosporine-induced cell death is triggered by a non-classical phosphoinositide 3-kinase s{hKl0ds
and does not require ERK activation. 2-&EkF4p'
Much of the current work on v3tJtb^'!
Discussion of these theories is beyond the scope of this review KyvZ?R
Based on the information contained in this 0XHQ5+"8
The result can be categorized into nine classes em$pU*`P
The results are illustrated by an example @ZKf3,J0
The experimental results for each xx time are reported in Table 2. eL!41_QI
From the results obtained so far, it seem that s27IeF3
Because of the inaccuracy of the ..., a conclusion cannot be drawn as ~%TWF+
Although much effort has been made to., this reality is far from completion. 0D X_*f
The results indicate that the total benefits are higher than the total costs. U$rMZk
Their results may then serve as guidelines for lower level models, less fuzzy and more H=t"qEp
detailed. s='+[*&&
Conclusion vW63j't_
From the discussion, one may conclude that ... r-2k<#^r
The first indication that .8hI
ad
In summary, we have shown that xNP_>Qa~
These results suggested that :FyF:=
Our findings have shown that >yIJ8IDF
Our observations have provided 3>,}N9P-v
This study reveals following five main findings: 1GY2aZ@
To our knowledge, this is the first immunohistochemical report of both PIM and HIFs in jYsAL=oh,*
the diabetic kidney. X1L@
G
As mentioned above, u8e_Lqx?
Moreover, we demonstrate that, )P$(]{
A number of studies have elaborated a body of knowledge about DE?v'7cmA
There are findings to indicate that p!.~hw9
On a descriptive level, there is agreement that for xad`-
vw
One issue that became particularly evident from our study is gcLz}84
Our data are in agreement with the findings and models presented by previous yD
WIflP0;
publications, in particular those of Jones et al FLZ9pb[T
According to our observations, i`,FXF)
…was not evident from our data ]@q%dsz
25 Lu.zc='\
It could therefore be concluded that lQA5HzC\
our data illustrate that %36x'Dn?
All these different aspects, as listed above, lead to &&ioGy}1
In fact, it has already been speculated that :
t
?B)
It should be furthermore mentioned in this context that 5/H,UL
To identify and verify the essential factors triggering developmental renin expression, it 7y=>Wa ?T[
is of interest now to study the development of intrarenal renin expression in mice with M .#}
defined gene mutations, using the results of this study as a reference system. lU
WXXuO]
In conclusion, using several approaches, we demonstrated that ^:nc'C gP
But we have not provided formal proof yet that |diI(2w
In summary, our results suggest that =KRM`_QShg
In conclusion, we identify OG\i?N
A better understanding of such regulatory systems may yield novel therapeutic [m~J6WB
approaches to glomerular diseases. wjarQog5Y
Preliminary results of short-term studies (2 weeks) indicate that dBWny&
However, we believe that nD
wh
However, despite our extensive measurements of these parameters, we found no :uL<UD,vu3
relationship between response and initial proteinuria, time to B-cell recovery, the ZR
.k'
development of antibodies to rituximab, B-cell count in the kidney tissue, nor the degree 16NHzAQ
of tubular interstitial damage present. A clear conclusion for efficacy, however, cannot be .?>Cav9:
obtained from an uncontrolled study, and definitive claims of efficacy should be reserved MlO OB
for rigorous, prospective, controlled, and randomized trials with the use of rituximab and }$6L]
that will also look for factors that may determine its efficacy in IMN. ~uQ*u.wi
It has become increasingly clear that Pz4#>tP
Form the above discussion, the conclusion can be reached that PN*
.9;5Z
The conclusions drawn are also valid gF6> /
In conclusion to this, it becomes obvious that the problem of xx lies not only in... 'u,|*o
We have attempted to introduce some concepts associated with a theory of bWPsfUn#
Considerable more work, hopefully, will be done in this area
)>a~ %~:
Employing the compositional rule of inference, the assessment of the xx compatibility in + NlnK6T/
achieving prescribed xx projectiles in any level of the hierarchy is made possible. <o E
Ay
This paper has presented a theoretical and experimental study of the xx process and xx +{UY9_~\3
concept. XM'tIE+|
The experimental research results will hopefully serve as useful feedback information for {kpad(E
improvements for xx work. n,I3\l9
The scope of this contribution was to introduce a xx method. %9-^,og
This has a tremendous impact on our understanding of {!,K[QwcI
Significant progress has been made in advancing RNAi therapeutics in a remarkably c3]t"TA,
short period of time InP E_
To date, little is known about… 1?@
HOu
Irrespective of the mechanisms and the molecules –which are still largely unknown – [ATJ!
O
involved in MSC functions, current data suggest that J[/WBVFDf
To better understand X [Y0r
26 %m&6'Rpfk
Future Research (常用于conclustion 的结尾) ^eW<-
n@^
Thus, first extension of the approach could be, EJ:O 1
Further studies are needed to determine the importance of 3%v)!dTa<^
Some improvements to the scheduling aspect of the model may be brought through :Cq73:1\B
additional levels in the hierarchy for more detailed representation of the scheduling
]b-2:M
activity. ;lObqs*?>
It also unveiled new aspects of the role played by thyroid hormone in response to injury 7q0_lEh
and tissue repair. =<]`'15"V
In the near future, the ongoing clinical trials with siRNAs for macular degeneration and d$IROZK-D
RSV may reveal the exciting potential of RNAi therapeutics as the next major class of (\\;A?
drug molecules. .}faWzRH9
In the next few years it should become clear whether ?N kKDvv
It should be an exciting time for researchers seeking to harness this powerful endogenous xHgC':l(0
pathway to treat human disease. ?Y"%BS+pt
Acknowledgement `[H^`
The authors are grateful to the insightful work provided by Mr. John W. Harney and all G,C`+1$*
of the fellows and students that contributed with some of the studies discussed in this %
/:1eE`!S
article ly[yn{
I would like to express my gratitude to all those who gave me the possibility to complete T//xxH]w-
this thesis. qbyYNlXqm
I want to thank the Department of Transport of the former Interim Government of }`=7%b`-?
Namibia for giving me permission to commence this thesis in the first instance, to do the w/@ZPBRo]
necessary research work and to use departmental data I`_2Q:r
This work was supported by NIH grants AI058695 and AI056900. We thank members of %q5dV<X'c
the laboratory and our collaborators for many useful suggestions. oh5'Isb$
We thank J. Maraganore and B. Greene for critical reading of the manuscript, M. SZGR9/*^
Duckman for graphics assistance and A. Capobianco for word processing assistance. Pe`mZCd^
Author contributions ?/|Xie
All authors were involved in experimental planning and data analysis and contributed to ^E}?YgNp
manuscript preparation. zP
nb_[YF
Tables and Figures KV0*dB;
Figure 7-1 sketches these relationships. ebA95v`Vms
The graphical representation of these functions is shown in Figure 1. X
}( s(6
The xx may be depicted as in Figure 1. U["0B8
Figure x shows the schematic diagram of the /r4l7K
Figure 1 though 2 provide a ... that $;5Q
mKQ'
the architecture of this expert system for .... is illustrated in Figure 2. {aAd (~YZ
Figure 2 gives the outline of an ... system UZ<K'H,q
Table shows the (zsG!v
27 L[<#>/NPy
Time-dependent changes of 3ijPm<wn
as shown in Table 1 and 2 _}R9!R0O
This concept is illustrated in Figure 2 ?R]`M_^&u!
At the top of Table xx are shown two blocks of data. (MLcA\LJ
Each table or matrix has constructs xx through xx as row;headings, "Z&.m..gc
xx through xx as column;headings. 4jXo5SkEJ
Time course of calculated renin immunoreactive tissue volumes during development of G*z\
^H
the mouse kidney. Data are single values per time point. They were derived from the \3hA_{ w
kidneys shown in Figure 1. {+9RJmZg
Yellow color indicates ?tYZ/
PCT3 cells were pretreated with 50 M PD98059 or 10 M U0126 for 30 min and then 4Ly>x>b<
cells were stimulated with 25 g ml-1 CsA for 6 h. ERK and PKB activation were o2
ng
analyzed by western blot using phospho-specific antibodies. v+LJx
ERK and PKB activation were measured by western blot using phospho-specific CH6 m
antibodies zT>!xGTu7~
The data are the means of three different experiments taking the number of untreated cells t5{P'v9J
(vehicle alone) as 100%. l~E~! MR
ERK and PKB activation were then analyzed by western blot with the corresponding b`X
''6
phospho-specific antibodies. To ensure equal amount of protein in each lane, western <>6j>w_|
blots against total protein were performed for ERK and PKB. nJe}U#
The means of optical density (OD) values of the bands detected in western blots of three f\_RW;y|m
different experiments are plotted, taking the maximal value of phosphorylation of ERK xFF!)k #
and PKB as 100%. LGt>=|=bj
CT3 cells were treated with 25 g ml-1 CsA or vehicle alone for 24 h and then fixed with 3y)\dln
paraformaldehyde and stained with Hoescht as indicated in Materials and Methods. Then dJR[9T_OF
cells were visualized by phase-contrast microscopy (upper panel) and fluorescence ?I=
1
T.
microscopy (lower panel). `=Ip>7T&
The data are the means of three experiments performed on different days taking the _V$'nz#>e
number of cells at the onset of the experiment as 100%. mGE!,!s}
Rats were killed at 4 h and at days 1, 2, 4, 7, 9, 14, 21, and 28 after disease induction t^'nh
1=
(n=9 each). `/sNX<mp
*P<0.05 versus non-nephritic rats. |<Ls;:5.
The expression of the CCN3 protein was detected by western blot analysis (n=4 each, PC,
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positive control, M, molecular weight markers). h~>1-T8
Original magnifications: 200 in A, E, G, H; 600 in B, C, D, F. G|,&V0*
Data are means s.d. of four independent experiments. >W> rhxU
*P<0.05 versus 0.5 h. h:fiUCw
PDGF-BB and -DD, but not PDGF-AA and -CC, induce a downregulation of CCN3
YNBM\Q
mRNA as determined by real-time RT-PCR and normalized to :UdW4N-
glyceraldehyde-3-phosphate dehydrogenase mRNA. ,1ev2T
Cell number was counted in duplicate using a Malassez hemocytometer =d8Rij-
Data are means s.d. of four independent experiments. *P<0.05 versus cells treated with wlNL;W@w
MCDB media for 24 h, #P<0.05. 58Ibje
28 J2qsZ
The curves are adjusted for history of coronary artery disease, CRP, and progression to ]({-vG\m
ESRD. _%Z.Re
Both models are independent of age, gender, baseline GFR and proteinuria, and other Aits<0
clinical characteristics and traditional and nontraditional cardiovascular risk factors VgbNZ{qk@
Correlation between serum bone-specific alkaline phosphatase (bAP) and serum PTH eC5*Q=ai,
(n=99, P<0.0001, r2=0.190) Ovt]3`U9J
Concentration of serum FGF23 concentration before starting and at the end of the 4-h F>\,`wP
hemodialysis procedure in 23 patients, expressed as log10 values (n=23, P<0.0001, Ii+3yE@c
Student's paired t-test). X
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Immunohistochemistry for the hypoxia marker pimonidazole (PIM) and HIF-2 ; UIEvwQ
arrow=endothelial cell and CD=collecting duct #czInXTTx
Magnification: 1000. [#GBn0BG)
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