18 h!Y?SO.b
Jurkat cells cultured in calcium-free RPMI-1640 medium (Gibco BRL; number 4 l1 i>_R
22300-107) containing calcium-free 10% FBS were triggered by anti-Fas IgM. The ~},~c:fF?
treated cells were harvested at the indicated time points and incubated with Fluo-3-AM at %YCd%lAe,
a final concentration of 1 micromolar for 30 min at 37°C (Scoltock et al., 2000). The iokP
mV
labeled cells (50,000 cells per treatment) were then analyzed by exciting the cells at 488 <A{y($
nm and examining the fluorescence emission of Fluo 3 at 530 nm with a FACS Scan, 6Hn)pD# U
(Becton Dickinson). A one micromolar concentration of LPA was used as a positive _uJVuCc
control for Ca2+ induction. The data thus obtained was analyzed with the software Win phS>T
MDI 2.8 and represented as contour plots.The effect of chelating intracellular calcium on LfllO
translocation of annexin I was studied by culturing Jurkat cells in the presence of 10 |!xpYT:
micromolar BAPTA-AM, with or without the addition of anti-Fas IgM. Cells were quRPg)
harvested and fractionated as detailed above, and the S-100 fractions were assessed by ENyAF%6
immunoblotting for presence or absence of annexin I. &SrGh$:X
Mouse bone marrow transduction and transplantation. FF3&Y^+^"
Retrovirus-mediated transduction of mouse bone marrow cells was done by published
A!4VjE>
methods49. Prestimulated low-density bone marrow cells were infected with high-titer 9fMSAB+c%
retrovirus supernatant on fibronectin-coated plates. Retrovirus supernatant was generated /sV?JV[t
in the phoenix-gp cells with a mouse stem cell virus–based retroviral vector coexpressing
p;e$kg1
EGFP and HA–RhoH as described50. EGFP+ sorted cells were transplanted by KqK9X
intravenous injection into the sublethally irradiated (300 rads with a 137Cs irradiator) T*R{L
Rag2-/- recipient mice. At 9 weeks after transplantation, thymus, peripheral blood, bone ;le0QA
Pf
marrow, spleen and lymph nodes from each recipient mouse were collected for analysis H@4/#V|Uy
of EGFP+ chimerism and hematopoietic lineage by flow cytometry. Expression of sNf
+ lga0
HA–RhoH and HA–RhoHF73F83 in EGFP+ sorted thymocytes of recipient mice was
aH
confirmed by immunoblot analysis. I5Q~T5Ar
Determination of renal morphology T
$IUKR
Kidney slices were postfixed in buffered 2% OsO4, dehydrated, and embedded in an VW
~Xbyf
Araldite-EM bed 812 mixture. Large sections were cut perpendicular to the renal capsule, +NT:<(;|i5
containing cortex, and medulla. Thin (1 m) sections were analyzed in a blinded manner cP J7E
for morphologic alterations, as previously detailed );!dg\U
Patient population 4]]b1^vVj
Patients included in the study met the following criteria: (1) biopsy-proven IMN; (2) =nw0# '
creatinine clearance 30 ml per min per 1.73 m2; and (3) persistent proteinuria >5 g per 4Lw'v: (
24 h despite treatment with an HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A) sjcQaF`=
reductase inhibitor, an ACEi, and/or ARB at maximal tolerated dose for at least 4 months. h)P]gT0f/
The Mayo Institutional Review Board and the Research Ethical Board, University Health (]}XLMi,|!
Network, University of Toronto approved the study protocol. All patients gave written j $KM9
informed consent. Patients who had been on treatment with prednisone, cyclosporine, or ^h`!f vyH
19 o3fR3P%$
mycophenolic mofetil within the last 4 months or alkylating agents within the last 6 0T=jR{j!o
months were not included in the study. Patients with active infection, diabetes, or a 9X.gg$P
secondary cause of MN (for example, hepatitis B, systemic lupus erythematosus (SLE), VGfD;8]z
medications, malignancies) were also excluded. <e[!3,%L
Treatment A
xRl*B
At enrollment, a low-sodium (<4 g day-1) and low-protein (0.8 g per kg per day of A7.JFf>
high-quality protein) diet was recommended and patients were encouraged to maintain 3b
,=
the same diet throughout the duration of the study. All patients received a similar cj`#Tg.
conservative treatment regimen that included loop diuretics to control edema, an #%;<FFu\
HMG-CoA reductase inhibitor, and an ACEi combined with an ARB if tolerated. Y7*'QKz2
-Blockers and non-dihydropyridine calcium channel blockers, in that order, were added LciL/?
when required to control systolic blood pressures to <135 mm Hg in >75% of the kqQphKkL
readings. Patients who after a minimum of 4 months of conservative therapy and kbD*=d}3{
maximized Ang II blockade had proteinuria >5 g per 24 h received two i.v. infusions of M^Q&A R'F
rituximab at a dose of 1000 mg on days 1 and 15. To minimize infusion reactions, 2LtU;}7s
patients were premedicated with acetaminophen (1000 mg) and diphenhydramine *;>V2!N=U
hydrochloride (50 mg) orally. In addition, methylprednisolone (100 mg, i.v.) was given XB6N[E
prior to the first rituximab infusion. B-cell depletion was defined as CD19+ count D>psh-,1
<5 cells per l at any time and B-cell recovery was defined as CD19+ cell count ]JkpR aP$
>15 cells per l. Patients treated with rituximab, who at month 6 had proteinuria >3 g per 2O}s*C$Xav
24 h and in whom CD19+ B-cell counts had increased to >15 cells per l, received a Lw!?T(SK
second course of rituximab treatment following the same protocol described above. bjU 2UcI"<
Follow-up 5d)'`hACe
In all patients, clinical and laboratory parameters including complete blood counts, QwKky ^A
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum n0Qh9
*h
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry ~{52JeUc P
and at months 1, 3, 6, 9, and 12. Creatinine clearance and protein and creatinine excretion qk"oFP6
in the urine were assessed by performing two consecutive 24-h urine collections at each ^bZ<9}
time point. Data were considered accurate when urinary creatinine excretion was z7NGpA(
consistent with a complete 24 h collection. The mean of the two measurements was 0KWy?6 X
considered for the analysis. The presence of HACAs was evaluated at baseline and at mcAH1k e
months 3, 6, 9, and 12. ="'rH.n #
Method / Approach / Study/ Technique < -@,
A discussion is presented of a problem-solving system PV Q#>_~5
To improve the efficiency of the method, the following approach may be applied. dQWA"6?i
In order to an investigation was made to find the causes of the b.w(x*a
Although large collections of rules and equations have been complied, none are generally c (_oK ?
accepted : .FfE
20 V29S*
This approach will be explained and discussed thoroughly in the body of the report. cLN[o8ZU
This can be accomplished by f:JYG]E &
This algorithm to compute the total cost can be described step by step as follows: `o^;fcnG
The above preliminary analysis has provided important information mPw56>
Various methods have been proposed for selecting an optimum... l/y
Kc8^<
These concepts have been applied to ;s!ns N
On the basis of the concept mentioned above, aQym=
6%e
This can be achieved by j4
&
In addition, tissues were stained for infiltrating lymphocytes (CD20 and CD3), and the cQ8$,fo
amount of interstitial fibrosis was quantified by histomorphometry. Formalin-fixed, P*"AtZuY]
paraffin-embedded sections were cut onto coated glass slides. Following heat-induced _sf0{/< )
antigen retrieval, sections were incubated at 20°C overnight with either anti-CD20 Zc' >}X[G
primary antibody or anti-CD3 primary antibody, both at 1:1000 dilution (Dako, Canada v[3hnLN%
Inc, Mississauga, Ontario, Canada). After rinsing all sections, pretreatment with 3% r{6B+3J
hydrogen peroxide was performed to prevent endogenous peroxidase activation. Sections G~iYF(:&
were incubated with a secondary rabbit anti-mouse antibody linked with avidin–biotin p#8LQP~0$
complex. Sections were counterstained with hematoxylin and examined by light a*n%SUP
microscopy. ,ovv
The HACA assay is a proprietary bridging enzyme-linked immunosorbent assay fE7a]REK
performed at Genentech Inc. that measures the antibody response to rituximab in human dc@wf;
o
serum samples. |uBot#K|
In all patients, clinical and laboratory parameters including complete blood counts, .V )2Tz
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum !k??Kj
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry }AH|~3|D
and at months 1, 3, 6, 9, and 12. Sy0s`\[
This fact suggests that a new concept
rgvc5p
This was accomplished by taking ... =iN_Ug+
The preparatory stage is very time consuming process. #O974f8
Test are performed for validity, completeness, and compatibility Ud(d Wj-/
There is little hope of achieving successful ... aFc'_FrQ
There has been an increasing awareness of the potential of using most ..so far made have 96V, [-arf
not taken this approach, with the exception of = {DB
Only a few studies can be found. #?d>S;)+
It is a very tedious process to go through x)sDf!d4bi
It is only when .. has been completed that .. may be effected 5lA 8e
The entire interpretation process is conducted in one's head. ez9M]! 8Lt
These approaches are sometimes very tedious. Z5^UF2`Q
Several techniques can be used uRcuy/CY
A polynomial parametric model can be written as [the following]/[follows]: "'dC>7* <
A xx model is constructed/formulated using xx. (Jr;:[4XC
A xx model represents an xx by its xx. Fi=8B&j
A process decision model captures the logic essential to \z?;6A
From the equation above, xx is equal to the summation of xx times the ... VrL>0d&d
21 J_Xf:Mz-
The validity of a xx model can be checked using Euler's formula. ,
&f20o
Given a model, one can mathematically determine whether ... or ... q&S
.C9W
Equations for xx need to be derived and implemented in the system. ?S7:KnU>K
A number of heuristic rules have been developed for &otgN<H9
Optimum .. techniques can be made more reliable by ... so that .`=PE&xq
An algorithm based on the characteristic ... is used to determine ;lt;]7
Euler's formula states the following: Mo=-P2)>lt
The completed model should agree with the formula. ~ +$><qj
For manufacturing purposes, a detailed and precise model of the object is necessary 6%y: hLT
Engineering design models are very well defined; therefore, \kO_"{7n
To keep the domain narrow enough to be implementable, yet wide enough to be useful. ' .<"jZ
Point of View ?P]md9$(+e
from an implementation standpoint, q|2C>{8
From the point of view of this application, *(IO<KAg8
From this point of view, Zadeh suggested an inference rule named xxx (CRI for short). A*i_-;W)
Information is the meaningful interpretation and correlation of some aggregation of data sva$@y7b
in order to allow one to make decisions. L
*@>/N
From a practical point of view, the computational aspects of an FLC require a Yl=
|P`
simplification of the fuzzy control algorithm. S_6
;e|
The use of a hammer to insert screws, although partly effective, tends to distort, destroy, NQD b;5:
and generally defeat the purpose of using a screw [Kusiak AI Implications for CIM g5[3[Z(.
p.129] ;0E4S
Statistical analysis 5.
i;IOx
Data were analyzed by one-way analysis of variance comparing the three conditions D bJ(N h
(sham operation, ischemic AKI, and bilateral nephrectomy) at each time point. If VPbNLi
significant F-statistic from analysis of variance existed, this test was followed by Dunnett 1[ Pbsb
post hoc multiple comparison procedure with sham operation as the control group. For all @pz2}Hd|
other comparisons, Student's t-test was used. A P-value of <0.05 was considered as ?`$4ZDM
statistically significant. "'L
SLp
Values are expressed as means s.e.m. and significance was evaluated by Mann–Whitney &s5*akG
U test using GraphPad Prism, version 4.0 software (GraphPad Software Inc., San Diego, G1|:b-C
CA, USA). Tj:F Qnx
All values are expressed as means s.d. Statistical significance (defined as P<0.05) was zZ63
P
evaluated using analysis of variance and Bonferroni t-tests, and the two-tailed Pearson's "cx" d:
test, where appropriate. 04:Dbt~=?p
Data are expressed as mean s.d., median and interquartile range, or frequencies, as QB3AL;7
appropriate. Variables who deviated from the normal distribution (positively skewed) ORN
E>6J
H
were log-transformed (log10) before the correlation study. Sk-Ti\
Data are represented as means s.e.m. Student's t-test and multiple comparisons with t-test tq[C"| dH
post hoc analysis of variance were used as indicated below, for the comparison of iVRz
22 `{%*DHa
morphological, immunohistological and functional parameters. Statistical significance i
BQf tq7
was set at P<0.05. S1."2AxO
The primary efficacy parameter was defined as change in urinary protein excretion from HhO$`YZ%>
baseline (week 0) to 12 months after treatment. The 12-month changes were tested p\ASf
against zero using the paired t-test. Secondary end points included 6-month changes in ^VQiq7 xm
protein; the number with PR or CR at 6 or 12 months; and changes in glomerular +x1sV *S
filtration rate (GFR), serum albumin, and lipid profiles. Study sample size was based on ;iEqa"gO
the desire to have 80% power to detect a drop in urinary protein of at least 2.0 g day-1. 2
UgjH
Assuming a two-sided hypothesis test performed at a significance level of 0.05 and an s.d. V}( "8L
of urinary protein change of 2.5 g, it was determined that 15 patients were required.2 (L\tp>
E-
Definition of remission status is according to the criteria established by Cattran et al.30 Rj8l]m6U9
CR was defined as proteinuria <0.3 g per 24 h, PR as proteinuria 3 g per 24 h, and a _T1e##Sq,
>50% reduction in peak proteinuria and non-response as <50% reduction in peak ixp %aRRP
proteinuria. Any patient reaching a CR or PR was considered a treatment success }.Ug`7%G
The statistical significance of differences for the mean values of cytokine concentration R75sK(oS
and T cell proliferation was determined with Student's t-test. Differences with a P value TnC'<zm9!
of less than 0.05 were considered significant. tIg_cY_y
Statistical significance was determined by Student's t-test. Data with a P value of less cD 1p5U
than 0.05 were considered significant. aW"BN 5eM>
In vitro and in vivo experiments were analyzed by the two-tailed Student's t-test, with P % $J^dF_0
values less than 0.05 considered statistically significant. u}pLO9V"`
The significance of differences among groups was determined by Student's t-test and 'RPe5 vB
one-way analysis of variance (SigmaStat software; Jandel). !$O +M#
Comparisons Z*k}I{0,-
As well, the pros and cons of these representations from a process planning point of view S -im
o
will be discussed. RxAZ<8T_
The method of using xx to implement xx described by Zadeh (1973) appeared more
w*aKb
suitable _- {
> e
As discussed [in the previous section]/[preciously], }4$UlTA'
Relation l{kum2DT
We can not invert F' directly because it defines a many-to-one mapping. 1;>J9
The relationships appear very complicate Ug/b;( dJ'
Lifting tasks involve complex and imprecise relationship between the task variables and Z[ys>\_To
the human operator's characteristics. y<kW2<?
These methods are based on the relationship between ... and ... E-CZk_K9
The fundamental concept of a fuzzy rating language is that we can establish a relationship p?ICZg:
among terms such as high, medium, and low, and then modify these relationships. 2WqjNqx)6
This article will thus mention the latter as well as the former. a8NL
The former two bear a close relation to a fuzzy Cartesian product. L~>pSP^a
Importance
y
m^
) &iez{[O
23
<<4U:
The emphasis is on an implementation of a general approach to rule based decision "&W80,O3
making. 0w)Gb}o$
Consideration / Attention -Q9} gaH_
Careful evaluation is necessary to ensure _TEjB:9eY
Such a formulation does not change further considerations. b@`h]]~:
Considerable attention has been paid to }hRw{#*8
Attention should be paid to an important finding of this investigation. O%}?DiSl
Caution should be exercised in this process to avoid ... 4h~CDy%_
Primary consideration is given to ... components, though others can be accommodated ZD ~ra7
After ... has been defined by ..., a carefully analysis is carried out/performed to determine j]P'xrWl]8
A number of factors such as ...need to be taken into consideration before making the -?z#
appropriate decision. $Y`oqw?g+^
It should be noted that 37<^Oly!
It is important to point out that ... mUA!GzJ~u-
These considerations have heightened interest in the possibility of providing ... u'`eCrKT*
We should stress the fundamental importance of the xx AdD,94/
Results. '9u(9S
Advantages / Disadvantage R`KlG/Tk
One of the major advantages of this new measure of xx is that it can be applied to the Y#[xX2z9
experimental study of *(E]]8o
One advantage of using a .. is the ease of preparing it. 9x`1VR
:
It has a very fast decision making process k:[T
#/;
All the algorithms involve mostly logical operations. 4v.{C"M
It can be easily and without additional cost implemented in a microprocessor;based r(j :C%?}C
environment. hh!4DHv
It can reduce the waste of designing from scratch. i"#pk"@`
The advantages of using a xx to represent xx are the following: [>D5(O
However, xx is not without its shortcomings. H
S)$|m_
In most cases, the xxx shows an improvement over the existing xxx. 2Q%*`
vCuV
Compared to the existing xx, the impacts of the xx are generally reduced by 5% to 9%. n|fKwWB\
The "best case" results shows a savings of 6% to 9%. m*|G2
Most of the existing works based on xx approach can only recognize a xx . Dy&{PeE!
Most of the above methods are computational expansive and limited to xx. ;Wgkf_3
Some other advantages of xx are the following: oT0:Ny
The problem is the limitation of this method to a limited domain of parts. c
HC1l
It proved limited in application because it demanded precision in system modeling that v:"Y
was impossible in practice. ; GEr8_7
There are advantages to be gained in the structuring of costs and benefits, the use of xx, G/ H>M%M
The disadvantages of this method are also disadvantages of conventional xx approaches. ?)g [Xc;K
This combines the best features of both techniques @6M>x=n5
24 }{}?mQ
Hopefully, this tool can be as the reference framework of for developing a xx platform, :w]NN\
and helping the administration, marketing, and knowledge management activities in r 6&+pSA>
virtual communities. h.\p+Qw.
Results !yvw5As %
An improvement on the result shown above can be made by based on the data provided nub!*)q
Recent work has identified 5S[:;o
Time-dependent changes of >`^;h]Q
Cyclosporine-induced cell death is triggered by a non-classical phosphoinositide 3-kinase [b3$em<^JV
and does not require ERK activation. Bjh8uW
G
Much of the current work on G.r=fNP
Discussion of these theories is beyond the scope of this review !C(PfsrR/
Based on the information contained in this qT0
1@Bku
The result can be categorized into nine classes gH.$B'
The results are illustrated by an example lk[u
The experimental results for each xx time are reported in Table 2. vsPIvW!V
From the results obtained so far, it seem that qoZe<jW (
Because of the inaccuracy of the ..., a conclusion cannot be drawn as E2tUL#
Although much effort has been made to., this reality is far from completion. >}/T&S
The results indicate that the total benefits are higher than the total costs. Q[FDk63;w
Their results may then serve as guidelines for lower level models, less fuzzy and more 2k
m0
detailed. BjHp3-A'
Conclusion 7q{yLcC"
From the discussion, one may conclude that ... <tm=
The first indication that a@AIv"q
In summary, we have shown that 'e6J
&X
These results suggested that O]?\<&y
Our findings have shown that flz7{W
Our observations have provided p@h<u!rL8
This study reveals following five main findings: mRt/d
To our knowledge, this is the first immunohistochemical report of both PIM and HIFs in ,|]k4F
the diabetic kidney. gQ|?~hYYv
As mentioned above, -al
Moreover, we demonstrate that, b8[
ayy
A number of studies have elaborated a body of knowledge about 7mn&w$MS4:
There are findings to indicate that 4^9qs%&
On a descriptive level, there is agreement that for 9JeT1\VvHY
One issue that became particularly evident from our study is nITkgN:s
Our data are in agreement with the findings and models presented by previous +i
}uRO
publications, in particular those of Jones et al hM&VMa [
According to our observations, M5xMTP-
…was not evident from our data )%
gU
25 tVNFulcz$
It could therefore be concluded that T t_QAIl
our data illustrate that Gs4t6+Al
All these different aspects, as listed above, lead to IU"n`HS
In fact, it has already been speculated that @Y ?p-&
It should be furthermore mentioned in this context that M"Z/E>ne
To identify and verify the essential factors triggering developmental renin expression, it 6aWNLJ
@
is of interest now to study the development of intrarenal renin expression in mice with H1
i+j;RN
defined gene mutations, using the results of this study as a reference system. 7c<_j55(
In conclusion, using several approaches, we demonstrated that XAB/S8 e
But we have not provided formal proof yet that @mcP-
In summary, our results suggest that ZfS"
In conclusion, we identify Smd83W&
A better understanding of such regulatory systems may yield novel therapeutic Qe]&
approaches to glomerular diseases. 3'7X[{uBr
Preliminary results of short-term studies (2 weeks) indicate that $SzuUI
However, we believe that S@ItgG?
X
However, despite our extensive measurements of these parameters, we found no )>pIAYCVP
relationship between response and initial proteinuria, time to B-cell recovery, the DRRy5+,I
development of antibodies to rituximab, B-cell count in the kidney tissue, nor the degree Q:]v4/MT
of tubular interstitial damage present. A clear conclusion for efficacy, however, cannot be `Tr !Gj_
obtained from an uncontrolled study, and definitive claims of efficacy should be reserved i}C%`1+(
for rigorous, prospective, controlled, and randomized trials with the use of rituximab and ,!98VJmr
that will also look for factors that may determine its efficacy in IMN. Wf&i{3z[
It has become increasingly clear that jVSU]LU E
Form the above discussion, the conclusion can be reached that ZRUI';5x
The conclusions drawn are also valid mvyOw
M
In conclusion to this, it becomes obvious that the problem of xx lies not only in... sb%l N
We have attempted to introduce some concepts associated with a theory of v2>Dn
=V
Considerable more work, hopefully, will be done in this area )u307Lg
Employing the compositional rule of inference, the assessment of the xx compatibility in }G<A$*L1
achieving prescribed xx projectiles in any level of the hierarchy is made possible. +F^X1
This paper has presented a theoretical and experimental study of the xx process and xx lH>6;sE
concept. ^;cJjl'=
The experimental research results will hopefully serve as useful feedback information for D!bi>]Yd
improvements for xx work. ?M04 cvm
The scope of this contribution was to introduce a xx method. <##aD3)
This has a tremendous impact on our understanding of ?$4CgN-
Significant progress has been made in advancing RNAi therapeutics in a remarkably %?z;'Y7D
short period of time kk/vgte-)e
To date, little is known about… bpfSe
Irrespective of the mechanisms and the molecules –which are still largely unknown – s;:quM
involved in MSC functions, current data suggest that Jl89}Sf
To better understand aGmbB7[BZ
26 @%#!-wC-5
Future Research (常用于conclustion 的结尾) \J^xpR_0u
Thus, first extension of the approach could be, U,nQnD"!t&
Further studies are needed to determine the importance of K#
plSD^f=
Some improvements to the scheduling aspect of the model may be brought through 3hNb
?
additional levels in the hierarchy for more detailed representation of the scheduling yykyvy
activity. D{s4Bo-
It also unveiled new aspects of the role played by thyroid hormone in response to injury |n.ydyu`
and tissue repair. Z!qF0UDj
In the near future, the ongoing clinical trials with siRNAs for macular degeneration and M.}QXta
RSV may reveal the exciting potential of RNAi therapeutics as the next major class of Gh9dv|m=[;
drug molecules. w^QqYUL${
In the next few years it should become clear whether G3t
4$3|
It should be an exciting time for researchers seeking to harness this powerful endogenous K?aUIkVs
pathway to treat human disease. 5V/CYcO
Acknowledgement N
bW5a3=
The authors are grateful to the insightful work provided by Mr. John W. Harney and all ^o*$+DbC
of the fellows and students that contributed with some of the studies discussed in this yj{:%Km:`
article {p@uj_pS
I would like to express my gratitude to all those who gave me the possibility to complete &;k`3`MC~w
this thesis. [u2)kH$
I want to thank the Department of Transport of the former Interim Government of ?YbZVoD)J
Namibia for giving me permission to commence this thesis in the first instance, to do the ;M Z@2CO
necessary research work and to use departmental data iP~dH/B|v
This work was supported by NIH grants AI058695 and AI056900. We thank members of _Uz}z#jt
the laboratory and our collaborators for many useful suggestions. :j2_Jn4UP
We thank J. Maraganore and B. Greene for critical reading of the manuscript, M. tY=n("=2
Duckman for graphics assistance and A. Capobianco for word processing assistance. d\ Z#XzI8
Author contributions g#ONtY@*U
All authors were involved in experimental planning and data analysis and contributed to H;n(qBSB
manuscript preparation. z[wk-a+w
Tables and Figures dj76YK
Figure 7-1 sketches these relationships. Up$vBE8i]
The graphical representation of these functions is shown in Figure 1. 4&/-xg87(
The xx may be depicted as in Figure 1. }f6x>
Figure x shows the schematic diagram of the P( W8XC
Figure 1 though 2 provide a ... that <ZNa`
the architecture of this expert system for .... is illustrated in Figure 2. \(LHcvbb
Figure 2 gives the outline of an ... system O!uB|*
Table shows the T8FKa4ikn
27 6P;1I+5m{q
Time-dependent changes of !+H=e>Y6
as shown in Table 1 and 2 UgJ^
NF2w
This concept is illustrated in Figure 2 l(zkMR$b8
At the top of Table xx are shown two blocks of data. N_r*Ig
Each table or matrix has constructs xx through xx as row;headings, N[ArwV2O
xx through xx as column;headings. *(d^k;
Time course of calculated renin immunoreactive tissue volumes during development of -ij1%#t z
the mouse kidney. Data are single values per time point. They were derived from the 2#1FI0,Pa*
kidneys shown in Figure 1. &H5
6mL{
Yellow color indicates Z/uRz]Hi
PCT3 cells were pretreated with 50 M PD98059 or 10 M U0126 for 30 min and then sS(^7GARa
cells were stimulated with 25 g ml-1 CsA for 6 h. ERK and PKB activation were [dU/;Sk5
analyzed by western blot using phospho-specific antibodies. +Mm0bqNN
ERK and PKB activation were measured by western blot using phospho-specific WM=kr$/3
antibodies +bf%]
The data are the means of three different experiments taking the number of untreated cells N'm:V
(vehicle alone) as 100%. =AcbX_[
ERK and PKB activation were then analyzed by western blot with the corresponding (%+DE4?
phospho-specific antibodies. To ensure equal amount of protein in each lane, western &<^@/osi
blots against total protein were performed for ERK and PKB. V`fh,(:
The means of optical density (OD) values of the bands detected in western blots of three ]r"{G*1Q
9
different experiments are plotted, taking the maximal value of phosphorylation of ERK g\*gHHa
and PKB as 100%. 6/#= dv
CT3 cells were treated with 25 g ml-1 CsA or vehicle alone for 24 h and then fixed with GF/p|I D
paraformaldehyde and stained with Hoescht as indicated in Materials and Methods. Then Z p_
_
cells were visualized by phase-contrast microscopy (upper panel) and fluorescence 06Hn:IT18
microscopy (lower panel). Bxa],inuZ
The data are the means of three experiments performed on different days taking the G0]n4"~+?
number of cells at the onset of the experiment as 100%. g:o\ r
(
Rats were killed at 4 h and at days 1, 2, 4, 7, 9, 14, 21, and 28 after disease induction C,IN+@
(n=9 each). ~l}TlRqL
*P<0.05 versus non-nephritic rats. E$ngmm[
The expression of the CCN3 protein was detected by western blot analysis (n=4 each, PC, k Lv_P[I
positive control, M, molecular weight markers). $&e(V6A@
Original magnifications: 200 in A, E, G, H; 600 in B, C, D, F. Zi[)(agAT
Data are means s.d. of four independent experiments. 1I@4xC
#X
*P<0.05 versus 0.5 h. YwF6/JA0^
PDGF-BB and -DD, but not PDGF-AA and -CC, induce a downregulation of CCN3 !gk\h
mRNA as determined by real-time RT-PCR and normalized to .nSupTyG
glyceraldehyde-3-phosphate dehydrogenase mRNA. iZSSd{jO
Cell number was counted in duplicate using a Malassez hemocytometer Ov<3?)ok
Data are means s.d. of four independent experiments. *P<0.05 versus cells treated with `Eu(r]:W
MCDB media for 24 h, #P<0.05. 8uGPyH
28 |,7J!7T(I
The curves are adjusted for history of coronary artery disease, CRP, and progression to (Dar6>!
ESRD. y(]|jRo
Both models are independent of age, gender, baseline GFR and proteinuria, and other z\E"={P&
clinical characteristics and traditional and nontraditional cardiovascular risk factors Gzir>'d2'V
Correlation between serum bone-specific alkaline phosphatase (bAP) and serum PTH $HXB !$d
(n=99, P<0.0001, r2=0.190) ~r!5d@f.6
Concentration of serum FGF23 concentration before starting and at the end of the 4-h a?Y1G3U'
hemodialysis procedure in 23 patients, expressed as log10 values (n=23, P<0.0001, O7d Fz)$
Student's paired t-test). O9|'8"AF
Immunohistochemistry for the hypoxia marker pimonidazole (PIM) and HIF-2 ; L@z !,r,
arrow=endothelial cell and CD=collecting duct %)/f; T6
Magnification: 1000. @gqw]_W
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