18 EJsb{$u
Jurkat cells cultured in calcium-free RPMI-1640 medium (Gibco BRL; number ny0`~bl{p
22300-107) containing calcium-free 10% FBS were triggered by anti-Fas IgM. The xC]/i(+bA
treated cells were harvested at the indicated time points and incubated with Fluo-3-AM at G#>nOB
a final concentration of 1 micromolar for 30 min at 37°C (Scoltock et al., 2000). The +yk>jx
labeled cells (50,000 cells per treatment) were then analyzed by exciting the cells at 488 USY^
[@o[f
nm and examining the fluorescence emission of Fluo 3 at 530 nm with a FACS Scan, Spb'jAKj'
(Becton Dickinson). A one micromolar concentration of LPA was used as a positive 6
kk(FVX
control for Ca2+ induction. The data thus obtained was analyzed with the software Win "=)`*"rr
MDI 2.8 and represented as contour plots.The effect of chelating intracellular calcium on n$8A"'.M
translocation of annexin I was studied by culturing Jurkat cells in the presence of 10 ">7 bnOJ
micromolar BAPTA-AM, with or without the addition of anti-Fas IgM. Cells were Cq=c'(cX
harvested and fractionated as detailed above, and the S-100 fractions were assessed by OmW|\d PU
immunoblotting for presence or absence of annexin I. wrWWXOZ4
Mouse bone marrow transduction and transplantation.
5 X rn]
Retrovirus-mediated transduction of mouse bone marrow cells was done by published BC3I{Y|
methods49. Prestimulated low-density bone marrow cells were infected with high-titer @W8RAS~
retrovirus supernatant on fibronectin-coated plates. Retrovirus supernatant was generated &F)lvtt|
in the phoenix-gp cells with a mouse stem cell virus–based retroviral vector coexpressing -c. a7
EGFP and HA–RhoH as described50. EGFP+ sorted cells were transplanted by NB[b[1 Ch
intravenous injection into the sublethally irradiated (300 rads with a 137Cs irradiator) %NuS!v>
Rag2-/- recipient mice. At 9 weeks after transplantation, thymus, peripheral blood, bone Lgy }Gm8u5
marrow, spleen and lymph nodes from each recipient mouse were collected for analysis 5,A/6b
of EGFP+ chimerism and hematopoietic lineage by flow cytometry. Expression of ZCa?uzeo]
HA–RhoH and HA–RhoHF73F83 in EGFP+ sorted thymocytes of recipient mice was gC?k6)p$N
confirmed by immunoblot analysis. /PO5z7n0J
Determination of renal morphology fz%u
rbJR
Kidney slices were postfixed in buffered 2% OsO4, dehydrated, and embedded in an w%htY.-
Araldite-EM bed 812 mixture. Large sections were cut perpendicular to the renal capsule, IP-mo!Y.
containing cortex, and medulla. Thin (1 m) sections were analyzed in a blinded manner z}Q54,9m
for morphologic alterations, as previously detailed z7 }@8F
Patient population (URWicaB
Patients included in the study met the following criteria: (1) biopsy-proven IMN; (2) >n#Pq{7aF
creatinine clearance 30 ml per min per 1.73 m2; and (3) persistent proteinuria >5 g per /.vB /{2
24 h despite treatment with an HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A) cu|{cy-
reductase inhibitor, an ACEi, and/or ARB at maximal tolerated dose for at least 4 months. I~Ziq10
The Mayo Institutional Review Board and the Research Ethical Board, University Health f>dWl$/_s
Network, University of Toronto approved the study protocol. All patients gave written Rc @p!Xi
informed consent. Patients who had been on treatment with prednisone, cyclosporine, or 8^2Q ~{i
19 =MA$xz3
mycophenolic mofetil within the last 4 months or alkylating agents within the last 6 qXgg"k%A\
months were not included in the study. Patients with active infection, diabetes, or a *?cE]U6;
secondary cause of MN (for example, hepatitis B, systemic lupus erythematosus (SLE), zl$'W=[rFs
medications, malignancies) were also excluded. ftRf~5d2
Treatment Y'wQ(6ok
At enrollment, a low-sodium (<4 g day-1) and low-protein (0.8 g per kg per day of 3nMXfh/
high-quality protein) diet was recommended and patients were encouraged to maintain !A qSG-
the same diet throughout the duration of the study. All patients received a similar Bg{"{poy
conservative treatment regimen that included loop diuretics to control edema, an Y<odXFIS
HMG-CoA reductase inhibitor, and an ACEi combined with an ARB if tolerated. #fd;]
-Blockers and non-dihydropyridine calcium channel blockers, in that order, were added .*+%-%CbP
when required to control systolic blood pressures to <135 mm Hg in >75% of the Z$"E|nRN
readings. Patients who after a minimum of 4 months of conservative therapy and eT0Y
p
maximized Ang II blockade had proteinuria >5 g per 24 h received two i.v. infusions of Bu$Z+o
rituximab at a dose of 1000 mg on days 1 and 15. To minimize infusion reactions, (Rj'd>%c
patients were premedicated with acetaminophen (1000 mg) and diphenhydramine >NDI<9<'0}
hydrochloride (50 mg) orally. In addition, methylprednisolone (100 mg, i.v.) was given $}+t|`*q8]
prior to the first rituximab infusion. B-cell depletion was defined as CD19+ count YBF|0A{[Y
<5 cells per l at any time and B-cell recovery was defined as CD19+ cell count ;hzm&My
>15 cells per l. Patients treated with rituximab, who at month 6 had proteinuria >3 g per -y]e`\+[
24 h and in whom CD19+ B-cell counts had increased to >15 cells per l, received a 3HCH-?U5
second course of rituximab treatment following the same protocol described above. $IUe](a{d
Follow-up JlaT
-j
In all patients, clinical and laboratory parameters including complete blood counts, rW),xfo0
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum f i#p('8
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry 'z\$.L
and at months 1, 3, 6, 9, and 12. Creatinine clearance and protein and creatinine excretion B\*"rSP\
in the urine were assessed by performing two consecutive 24-h urine collections at each A*b>@>2
time point. Data were considered accurate when urinary creatinine excretion was :^bjn3b
consistent with a complete 24 h collection. The mean of the two measurements was dkUh[yo"H
considered for the analysis. The presence of HACAs was evaluated at baseline and at jc0Trs{Jf
months 3, 6, 9, and 12. Wy:xiP
Method / Approach / Study/ Technique ;Gnk8lIsb
A discussion is presented of a problem-solving system L@VIC|~E
To improve the efficiency of the method, the following approach may be applied. <?h,;]U
In order to an investigation was made to find the causes of the o%j[]P@4G
Although large collections of rules and equations have been complied, none are generally w%' 8bH!
accepted p Xap<T
20 Tup2
;\y
This approach will be explained and discussed thoroughly in the body of the report. (Vz\02,K
This can be accomplished by )Up'W
This algorithm to compute the total cost can be described step by step as follows:
<h"07.y
The above preliminary analysis has provided important information :@oy5zib
Various methods have been proposed for selecting an optimum... **KkPjAO?
These concepts have been applied to wbImE;-Z
On the basis of the concept mentioned above, gR#lRA/
This can be achieved by jT:z#B%
In addition, tissues were stained for infiltrating lymphocytes (CD20 and CD3), and the
qPH=2k,H
amount of interstitial fibrosis was quantified by histomorphometry. Formalin-fixed, qZ=%ru
paraffin-embedded sections were cut onto coated glass slides. Following heat-induced 5,~Ju>y*
antigen retrieval, sections were incubated at 20°C overnight with either anti-CD20 vQ_D%
f4;
primary antibody or anti-CD3 primary antibody, both at 1:1000 dilution (Dako, Canada azKiXr#_(
Inc, Mississauga, Ontario, Canada). After rinsing all sections, pretreatment with 3% @gM>Lxj
hydrogen peroxide was performed to prevent endogenous peroxidase activation. Sections ba`V`0p- (
were incubated with a secondary rabbit anti-mouse antibody linked with avidin–biotin z]d2
rzV(_
complex. Sections were counterstained with hematoxylin and examined by light /&jh10}H
microscopy. xsN)a!
The HACA assay is a proprietary bridging enzyme-linked immunosorbent assay rAwuWM@BIg
performed at Genentech Inc. that measures the antibody response to rituximab in human Lbwc2Q,.-
serum samples. ?ypX``3#s7
In all patients, clinical and laboratory parameters including complete blood counts, -RK R.,
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum X8uAwHa6F
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry GFX$vn-/F
and at months 1, 3, 6, 9, and 12. PYhRP00}M
This fact suggests that a new concept qy\Z2k
This was accomplished by taking ... XhEJF !
The preparatory stage is very time consuming process. MUjfqxTT
Test are performed for validity, completeness, and compatibility *kM^l!<g
There is little hope of achieving successful ... l'm\*=3
There has been an increasing awareness of the potential of using most ..so far made have _7=LSf,9
not taken this approach, with the exception of 7AouiL 2-W
Only a few studies can be found. L,7+26XV"B
It is a very tedious process to go through mLb>*xt$b@
It is only when .. has been completed that .. may be effected 29tih{xx
The entire interpretation process is conducted in one's head. &cuDGo.
These approaches are sometimes very tedious. 40$- ]i
Several techniques can be used =,w(D~ps
A polynomial parametric model can be written as [the following]/[follows]: Wfp>BC
A xx model is constructed/formulated using xx. +rN&@}Jt.
A xx model represents an xx by its xx. .d;|iwl
A process decision model captures the logic essential to )1iqM]~;B
From the equation above, xx is equal to the summation of xx times the ... +Jl
PQ~5
21 /+JHnedK
The validity of a xx model can be checked using Euler's formula. zkb[u"
Given a model, one can mathematically determine whether ... or ... -POsbb>
Equations for xx need to be derived and implemented in the system. 9?38/2kX4
A number of heuristic rules have been developed for O]VHX![Y$
Optimum .. techniques can be made more reliable by ... so that ^SvGSxi
An algorithm based on the characteristic ... is used to determine G;he:Bf
Euler's formula states the following: d3jzGJrU}
The completed model should agree with the formula. \PL0-.t,
For manufacturing purposes, a detailed and precise model of the object is necessary YS0^!7u
Engineering design models are very well defined; therefore, VxW>XxG0
To keep the domain narrow enough to be implementable, yet wide enough to be useful. 34c+70x7
Point of View 9(pF!}1%\
from an implementation standpoint, (MzThGJK_
From the point of view of this application, oZ\zi> Y,
From this point of view, Zadeh suggested an inference rule named xxx (CRI for short). 1TlMB
Information is the meaningful interpretation and correlation of some aggregation of data w[]\%`69}Z
in order to allow one to make decisions. ZfVw33z
From a practical point of view, the computational aspects of an FLC require a 7-("ppYX=
simplification of the fuzzy control algorithm. <Gj]XAoe%
The use of a hammer to insert screws, although partly effective, tends to distort, destroy, &Pe[kCO]
and generally defeat the purpose of using a screw [Kusiak AI Implications for CIM b
ltZQI|
p.129] siz:YRur
Statistical analysis Nn{/_QG
Data were analyzed by one-way analysis of variance comparing the three conditions '#u=wyp
(sham operation, ischemic AKI, and bilateral nephrectomy) at each time point. If ;nw}x4Y[
significant F-statistic from analysis of variance existed, this test was followed by Dunnett ZsN3 MbY
post hoc multiple comparison procedure with sham operation as the control group. For all .(Tf$V
other comparisons, Student's t-test was used. A P-value of <0.05 was considered as `teaE7^Wm
statistically significant. pGP$2
Values are expressed as means s.e.m. and significance was evaluated by Mann–Whitney hx
hs>eY
U test using GraphPad Prism, version 4.0 software (GraphPad Software Inc., San Diego, Y@MFH>*
CA, USA). d=/a{lP\
All values are expressed as means s.d. Statistical significance (defined as P<0.05) was ,B$e'KQ
evaluated using analysis of variance and Bonferroni t-tests, and the two-tailed Pearson's J<-2dvq
test, where appropriate. 9&Ny;oy#6
Data are expressed as mean s.d., median and interquartile range, or frequencies, as FB
n .
4
appropriate. Variables who deviated from the normal distribution (positively skewed) `! _mIh}
were log-transformed (log10) before the correlation study. [-h=L
Jf#
Data are represented as means s.e.m. Student's t-test and multiple comparisons with t-test v2 }>/b)
post hoc analysis of variance were used as indicated below, for the comparison of bix}#M
22 zfK3$|
morphological, immunohistological and functional parameters. Statistical significance Y 016Xg5
was set at P<0.05. 7-c3^5gn{
The primary efficacy parameter was defined as change in urinary protein excretion from K!;>/3Y2-
baseline (week 0) to 12 months after treatment. The 12-month changes were tested e@L?jBj8m
against zero using the paired t-test. Secondary end points included 6-month changes in I=|}%WO#
protein; the number with PR or CR at 6 or 12 months; and changes in glomerular 0ae}!
LO
filtration rate (GFR), serum albumin, and lipid profiles. Study sample size was based on $-M'
the desire to have 80% power to detect a drop in urinary protein of at least 2.0 g day-1. -_fh=}.n+"
Assuming a two-sided hypothesis test performed at a significance level of 0.05 and an s.d. bf$4Z: Y
of urinary protein change of 2.5 g, it was determined that 15 patients were required.2 9FmX^t$T
Definition of remission status is according to the criteria established by Cattran et al.30 >?ckBU9
CR was defined as proteinuria <0.3 g per 24 h, PR as proteinuria 3 g per 24 h, and a ,dSP%?vV
>50% reduction in peak proteinuria and non-response as <50% reduction in peak 5db9C}0
proteinuria. Any patient reaching a CR or PR was considered a treatment success )X5en=[)O
The statistical significance of differences for the mean values of cytokine concentration kT4Oal+4
and T cell proliferation was determined with Student's t-test. Differences with a P value YE1X*'4
of less than 0.05 were considered significant. A9ru]|?
Statistical significance was determined by Student's t-test. Data with a P value of less }yMAs
than 0.05 were considered significant. ;p/$9b.0:
In vitro and in vivo experiments were analyzed by the two-tailed Student's t-test, with P jHBP:c
values less than 0.05 considered statistically significant. 5/4q}U3
The significance of differences among groups was determined by Student's t-test and AOR(1Qyo
one-way analysis of variance (SigmaStat software; Jandel). jt4c*0z
Comparisons w$z]Z-
As well, the pros and cons of these representations from a process planning point of view D7OPFN7`
will be discussed. z*OQ4_
The method of using xx to implement xx described by Zadeh (1973) appeared more ff.;6R\
suitable 'I1^70bB
As discussed [in the previous section]/[preciously], 0ZjinWkR[
Relation 7~7_T#dTh
We can not invert F' directly because it defines a many-to-one mapping. ^{
w]r5d
The relationships appear very complicate H?W8_XiN
Lifting tasks involve complex and imprecise relationship between the task variables and Uh8c!CA8:\
the human operator's characteristics. ?(P3ZTk?.
These methods are based on the relationship between ... and ... 3v7*@(y
The fundamental concept of a fuzzy rating language is that we can establish a relationship ,]qTJ`J
among terms such as high, medium, and low, and then modify these relationships. k\`~v$R3
This article will thus mention the latter as well as the former. '}.Z' %;
The former two bear a close relation to a fuzzy Cartesian product. xix:=
a
Importance euc|G Xs
) N)N\iad^
23 MW$
X4<*KD
The emphasis is on an implementation of a general approach to rule based decision Ww
Bs_OMc
making. hd)HJb-aR
Consideration / Attention wYIlp
Careful evaluation is necessary to ensure 380M&Guh
Such a formulation does not change further considerations. E ) iEWc
Considerable attention has been paid to pa)2TL/@
Attention should be paid to an important finding of this investigation. ,/bv3pE
Caution should be exercised in this process to avoid ... _BP!{~&;
Primary consideration is given to ... components, though others can be accommodated -U#e
After ... has been defined by ..., a carefully analysis is carried out/performed to determine oe*Y(T\G
A number of factors such as ...need to be taken into consideration before making the b6y/o48
appropriate decision. "fTW2D74
It should be noted that
f7_EqS=(
It is important to point out that ... 7 _*k<W7|
These considerations have heightened interest in the possibility of providing ... AYnk.H-v
We should stress the fundamental importance of the xx r{pbUk
Results. WMw^zq?hd@
Advantages / Disadvantage y.PsC '
One of the major advantages of this new measure of xx is that it can be applied to the #Z)8,N
experimental study of 'W2B**}
One advantage of using a .. is the ease of preparing it. 2Sg^SZFH+o
It has a very fast decision making process v3q.,I_
All the algorithms involve mostly logical operations. }mx>3G{d
It can be easily and without additional cost implemented in a microprocessor;based jf
WZLb)
environment. T4lE-g2%M
It can reduce the waste of designing from scratch.
e#/SFI0m
The advantages of using a xx to represent xx are the following: jM%8h$&E
However, xx is not without its shortcomings. zlhHSy K
In most cases, the xxx shows an improvement over the existing xxx. 2iV/?.<Z&
Compared to the existing xx, the impacts of the xx are generally reduced by 5% to 9%. ]?3-;D.eG
The "best case" results shows a savings of 6% to 9%. lc8zF5
Most of the existing works based on xx approach can only recognize a xx . u>y/<9]q8
Most of the above methods are computational expansive and limited to xx. S+*g
Some other advantages of xx are the following: EjDr
The problem is the limitation of this method to a limited domain of parts. 8[\~}Q6
It proved limited in application because it demanded precision in system modeling that 4;D>s8dgG
was impossible in practice. hqwDlapTt
There are advantages to be gained in the structuring of costs and benefits, the use of xx, > %B7/l$
The disadvantages of this method are also disadvantages of conventional xx approaches. p/WE[8U
This combines the best features of both techniques oH>G3n|U^
24 .jK,6't^
Hopefully, this tool can be as the reference framework of for developing a xx platform, U3OXO1
and helping the administration, marketing, and knowledge management activities in 5kdh!qy[$,
virtual communities. |:b!e
Results ]rMHO
An improvement on the result shown above can be made by based on the data provided $dkkgsw7
Recent work has identified s.j6"
Q[W
Time-dependent changes of /Ps5Og
Cyclosporine-induced cell death is triggered by a non-classical phosphoinositide 3-kinase >yr1wVS
and does not require ERK activation. </-aG[Fi
Much of the current work on ]3&BLq
Discussion of these theories is beyond the scope of this review ipH'}~=ID
Based on the information contained in this e{E8_2d
The result can be categorized into nine classes :/~TV
The results are illustrated by an example \I+#M-V
The experimental results for each xx time are reported in Table 2. W`g zMx
From the results obtained so far, it seem that :=e"D;5
Because of the inaccuracy of the ..., a conclusion cannot be drawn as G-Dc(QhU&
Although much effort has been made to., this reality is far from completion. Dg'BlrwbR
The results indicate that the total benefits are higher than the total costs. 9?.
Their results may then serve as guidelines for lower level models, less fuzzy and more 5"mH6%d :8
detailed. Y |'}VU
Conclusion 0.z\YTZ9
From the discussion, one may conclude that ... {2T;^+KE
The first indication that R>y/Y<5=
In summary, we have shown that vNSeNS@jxC
These results suggested that |?<^4U8
Our findings have shown that v&r\Z @%
Our observations have provided fp?cb2'7
This study reveals following five main findings: -W.bOr
To our knowledge, this is the first immunohistochemical report of both PIM and HIFs in iCNJ%AZH
the diabetic kidney. `L7Cf&W\l8
As mentioned above, SmCtwcB1
Moreover, we demonstrate that, Tu&W7aoX5
A number of studies have elaborated a body of knowledge about nXAGwU8a
There are findings to indicate that Rg%Xy`gS
On a descriptive level, there is agreement that for \W+Hzf]
W#
One issue that became particularly evident from our study is >gf,8flgj
Our data are in agreement with the findings and models presented by previous q$7/X;A
publications, in particular those of Jones et al gPcOm
b
According to our observations, X]c>clk
,
…was not evident from our data ZDbzH=[
25 f[Xsri
It could therefore be concluded that iH>IV0
<
our data illustrate that BY>]6SrP
All these different aspects, as listed above, lead to VYnB&3%DF
In fact, it has already been speculated that cQrXrij;!
It should be furthermore mentioned in this context that Kj/Lcx;bh
To identify and verify the essential factors triggering developmental renin expression, it &/8B
(0<
is of interest now to study the development of intrarenal renin expression in mice with OXS.CFZM
defined gene mutations, using the results of this study as a reference system. =4
NKXP~C
In conclusion, using several approaches, we demonstrated that Qq3>Xv <
But we have not provided formal proof yet that 5#mHWBGd7
In summary, our results suggest that WXd#`f %
In conclusion, we identify ;E0x#JUrw
A better understanding of such regulatory systems may yield novel therapeutic J~)JsAXAI
approaches to glomerular diseases. py$i{v%
Preliminary results of short-term studies (2 weeks) indicate that EOXuc9>
G
However, we believe that }c=YiH,o
However, despite our extensive measurements of these parameters, we found no =yZ6 $ hK
relationship between response and initial proteinuria, time to B-cell recovery, the .t.4y.
97
development of antibodies to rituximab, B-cell count in the kidney tissue, nor the degree zvwv7JtB
of tubular interstitial damage present. A clear conclusion for efficacy, however, cannot be jQ['f\R
obtained from an uncontrolled study, and definitive claims of efficacy should be reserved x LGMN)@r
for rigorous, prospective, controlled, and randomized trials with the use of rituximab and ]L?WC
that will also look for factors that may determine its efficacy in IMN. 83F]d+n
It has become increasingly clear that Ct]?
/
Form the above discussion, the conclusion can be reached that m}t`43}QE
The conclusions drawn are also valid @B9#Hrc
In conclusion to this, it becomes obvious that the problem of xx lies not only in... O@>{%u
We have attempted to introduce some concepts associated with a theory of A_+*b
[P
Considerable more work, hopefully, will be done in this area u zL|yxt
Employing the compositional rule of inference, the assessment of the xx compatibility in }]. |7h
achieving prescribed xx projectiles in any level of the hierarchy is made possible. AjINO}b
This paper has presented a theoretical and experimental study of the xx process and xx 5Zl7crA [
concept. zwK$ q=-:
The experimental research results will hopefully serve as useful feedback information for c+jnQM'
improvements for xx work. ._<gc;G
The scope of this contribution was to introduce a xx method. GuQ#
This has a tremendous impact on our understanding of iBCIJ!;
Significant progress has been made in advancing RNAi therapeutics in a remarkably Jo9c|\4
short period of time BXr._y, cr
To date, little is known about… ]CgZt'h{
Irrespective of the mechanisms and the molecules –which are still largely unknown – #J%Fi).^)
involved in MSC functions, current data suggest that BF
U#FE)s
To better understand N~B'gJJDx
26 +
N!/>w]n
Future Research (常用于conclustion 的结尾) o
Hrx$>W]
Thus, first extension of the approach could be, VqeW;8&*iv
Further studies are needed to determine the importance of X($@E!|
Some improvements to the scheduling aspect of the model may be brought through
=[o/D0-Kn
additional levels in the hierarchy for more detailed representation of the scheduling 62#8c~dL
activity. IO?6F@(
It also unveiled new aspects of the role played by thyroid hormone in response to injury ni3A+Y0
and tissue repair. 92L{be;SY
In the near future, the ongoing clinical trials with siRNAs for macular degeneration and a4N8zDS
RSV may reveal the exciting potential of RNAi therapeutics as the next major class of znd fIt^
drug molecules. )X-TJ+d
In the next few years it should become clear whether H/O v8|
It should be an exciting time for researchers seeking to harness this powerful endogenous j(\jYH>
pathway to treat human disease. U+g<lgH1J
Acknowledgement >HQ<KFA
The authors are grateful to the insightful work provided by Mr. John W. Harney and all )UG<KcdI
of the fellows and students that contributed with some of the studies discussed in this k'+Mc%pg4E
article Toc="F`SW
I would like to express my gratitude to all those who gave me the possibility to complete ,]-A~ ^|
this thesis. =k22f`8ew
I want to thank the Department of Transport of the former Interim Government of d!]_n|B@9
Namibia for giving me permission to commence this thesis in the first instance, to do the QOY M/1U
necessary research work and to use departmental data k/V:QdD Sb
This work was supported by NIH grants AI058695 and AI056900. We thank members of Md ,KW#
the laboratory and our collaborators for many useful suggestions.
dzZ75
We thank J. Maraganore and B. Greene for critical reading of the manuscript, M. j
/=4f
Duckman for graphics assistance and A. Capobianco for word processing assistance. Se-
n#
Author contributions ]pP [0S
All authors were involved in experimental planning and data analysis and contributed to RP4P"m(
manuscript preparation. 'w5g s}1D
Tables and Figures o>Jr6:D(
Figure 7-1 sketches these relationships. LdM9k(
The graphical representation of these functions is shown in Figure 1. g}=opw6z
The xx may be depicted as in Figure 1. :=K <2
Figure x shows the schematic diagram of the t!,GI&
Figure 1 though 2 provide a ... that KcQe1mT!+
the architecture of this expert system for .... is illustrated in Figure 2. jVtRn.qh
Figure 2 gives the outline of an ... system fer~NlX
Table shows the $pg1Av7l
27 ZeyAbo
Time-dependent changes of 0'9zXJ"
as shown in Table 1 and 2 P@z,[,sy"$
This concept is illustrated in Figure 2 RIO4`,
At the top of Table xx are shown two blocks of data. ,[{)4J$MV
Each table or matrix has constructs xx through xx as row;headings, C{8d^SCA"
xx through xx as column;headings. h#>%\Pvt;
Time course of calculated renin immunoreactive tissue volumes during development of .oNs8._:
the mouse kidney. Data are single values per time point. They were derived from the ZOK,P
kidneys shown in Figure 1. eQRY
xx{
Yellow color indicates >$yqx1=jW
PCT3 cells were pretreated with 50 M PD98059 or 10 M U0126 for 30 min and then k7gm)}RKcu
cells were stimulated with 25 g ml-1 CsA for 6 h. ERK and PKB activation were e5 ?;{H
analyzed by western blot using phospho-specific antibodies.
/M1 /
ERK and PKB activation were measured by western blot using phospho-specific _K?{DnTb
antibodies `]8z]PD
The data are the means of three different experiments taking the number of untreated cells 0;kp`hB
(vehicle alone) as 100%. w<ol$2&B
ERK and PKB activation were then analyzed by western blot with the corresponding Z7X_U`Q
phospho-specific antibodies. To ensure equal amount of protein in each lane, western 5IPZ;
blots against total protein were performed for ERK and PKB. v{Rj,O
u
The means of optical density (OD) values of the bands detected in western blots of three xSQ:#o=8G
different experiments are plotted, taking the maximal value of phosphorylation of ERK {F/0pvP9
and PKB as 100%. h}k/okG
CT3 cells were treated with 25 g ml-1 CsA or vehicle alone for 24 h and then fixed with a}Dx"zl;
paraformaldehyde and stained with Hoescht as indicated in Materials and Methods. Then )NoNgU\7!
cells were visualized by phase-contrast microscopy (upper panel) and fluorescence oM&}akPE
microscopy (lower panel). f8T6(cA
The data are the means of three experiments performed on different days taking the atW=xn
number of cells at the onset of the experiment as 100%. (^:0g.~c
Rats were killed at 4 h and at days 1, 2, 4, 7, 9, 14, 21, and 28 after disease induction GI~JIXHTQ
(n=9 each). +ld]P}
*P<0.05 versus non-nephritic rats. [O"9OW'2!B
The expression of the CCN3 protein was detected by western blot analysis (n=4 each, PC, <}G*/ z?/
positive control, M, molecular weight markers). ig Q,ZY1
Original magnifications: 200 in A, E, G, H; 600 in B, C, D, F. V__|NVoOm
Data are means s.d. of four independent experiments. N-0kB vo
*P<0.05 versus 0.5 h. y@#JzfY?Hr
PDGF-BB and -DD, but not PDGF-AA and -CC, induce a downregulation of CCN3 (R.l{(A
mRNA as determined by real-time RT-PCR and normalized to |XDbf3^6
glyceraldehyde-3-phosphate dehydrogenase mRNA. N)kZ2|oD
Cell number was counted in duplicate using a Malassez hemocytometer F9Hxqa#1T
Data are means s.d. of four independent experiments. *P<0.05 versus cells treated with /IO<TF(X
MCDB media for 24 h, #P<0.05. [T6MaP?
28 ru#CywK{{;
The curves are adjusted for history of coronary artery disease, CRP, and progression to ,&q
Q[i
ESRD. aANzL
Both models are independent of age, gender, baseline GFR and proteinuria, and other KE_GC ;bQ
clinical characteristics and traditional and nontraditional cardiovascular risk factors h{Zd, 9H
Correlation between serum bone-specific alkaline phosphatase (bAP) and serum PTH |EjMpRNE
(n=99, P<0.0001, r2=0.190) p%]ZG,
Concentration of serum FGF23 concentration before starting and at the end of the 4-h UWPzRk#s"
hemodialysis procedure in 23 patients, expressed as log10 values (n=23, P<0.0001, DOa%|H'P
Student's paired t-test). *;0Ods+IcY
Immunohistochemistry for the hypoxia marker pimonidazole (PIM) and HIF-2 ; <0M2qt8
arrow=endothelial cell and CD=collecting duct px~ :'U
Magnification: 1000. ro8c-[
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