18 V$ZclV2:Ih
Jurkat cells cultured in calcium-free RPMI-1640 medium (Gibco BRL; number 97liSd
22300-107) containing calcium-free 10% FBS were triggered by anti-Fas IgM. The k;\gYb%L
treated cells were harvested at the indicated time points and incubated with Fluo-3-AM at q|*^{(tWs
a final concentration of 1 micromolar for 30 min at 37°C (Scoltock et al., 2000). The G&S2U=KdV%
labeled cells (50,000 cells per treatment) were then analyzed by exciting the cells at 488 $udhTI#,
nm and examining the fluorescence emission of Fluo 3 at 530 nm with a FACS Scan, W>#yXg9
(Becton Dickinson). A one micromolar concentration of LPA was used as a positive "Not /
8J
control for Ca2+ induction. The data thus obtained was analyzed with the software Win #@FA=p[%
MDI 2.8 and represented as contour plots.The effect of chelating intracellular calcium on <Yki8
translocation of annexin I was studied by culturing Jurkat cells in the presence of 10 Ki><~!L
micromolar BAPTA-AM, with or without the addition of anti-Fas IgM. Cells were v+LJx
harvested and fractionated as detailed above, and the S-100 fractions were assessed by 1<ag=D`F_"
immunoblotting for presence or absence of annexin I. 6*i**
Mouse bone marrow transduction and transplantation. l
5-[
a
Retrovirus-mediated transduction of mouse bone marrow cells was done by published ?=&; A
methods49. Prestimulated low-density bone marrow cells were infected with high-titer rpKZ>S|7+)
retrovirus supernatant on fibronectin-coated plates. Retrovirus supernatant was generated 6qT@M0)i
in the phoenix-gp cells with a mouse stem cell virus–based retroviral vector coexpressing 6:5K?Yo
EGFP and HA–RhoH as described50. EGFP+ sorted cells were transplanted by 'dwW~4|B
intravenous injection into the sublethally irradiated (300 rads with a 137Cs irradiator) C
@
XS
Rag2-/- recipient mice. At 9 weeks after transplantation, thymus, peripheral blood, bone s70Z&3A
marrow, spleen and lymph nodes from each recipient mouse were collected for analysis poBeEpbs
of EGFP+ chimerism and hematopoietic lineage by flow cytometry. Expression of x)C}
HA–RhoH and HA–RhoHF73F83 in EGFP+ sorted thymocytes of recipient mice was x^K4&'</
confirmed by immunoblot analysis. \\S
QACN
Determination of renal morphology 9[qOfIny
Kidney slices were postfixed in buffered 2% OsO4, dehydrated, and embedded in an ?I?G+(bq
Araldite-EM bed 812 mixture. Large sections were cut perpendicular to the renal capsule, ZeV@ X
containing cortex, and medulla. Thin (1 m) sections were analyzed in a blinded manner +RJ{)Nec
for morphologic alterations, as previously detailed XmQ;Roe
Patient population == wX.y\.n
Patients included in the study met the following criteria: (1) biopsy-proven IMN; (2) gC?}1]9c
creatinine clearance 30 ml per min per 1.73 m2; and (3) persistent proteinuria >5 g per hi,="
/9
24 h despite treatment with an HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A) \9[vi +T
reductase inhibitor, an ACEi, and/or ARB at maximal tolerated dose for at least 4 months. W^iK9|[qp
The Mayo Institutional Review Board and the Research Ethical Board, University Health ^{"i eVn
Network, University of Toronto approved the study protocol. All patients gave written eKr>>4,-P
informed consent. Patients who had been on treatment with prednisone, cyclosporine, or d[t0K]
19 ^8&}Nk[ j
mycophenolic mofetil within the last 4 months or alkylating agents within the last 6 UIEvwQ
months were not included in the study. Patients with active infection, diabetes, or a c=u+X`
Q
secondary cause of MN (for example, hepatitis B, systemic lupus erythematosus (SLE), ,g?ny<#o
medications, malignancies) were also excluded. Ubu&$4a
Treatment kZF]BPh.
At enrollment, a low-sodium (<4 g day-1) and low-protein (0.8 g per kg per day of w"0$cL3
high-quality protein) diet was recommended and patients were encouraged to maintain .6gx|V+
the same diet throughout the duration of the study. All patients received a similar P'VHga
conservative treatment regimen that included loop diuretics to control edema, an Vs5 &X+k
HMG-CoA reductase inhibitor, and an ACEi combined with an ARB if tolerated. "\*)KH`C
-Blockers and non-dihydropyridine calcium channel blockers, in that order, were added CRo'r/G
when required to control systolic blood pressures to <135 mm Hg in >75% of the =\]gL%N-|
readings. Patients who after a minimum of 4 months of conservative therapy and `:
lcN0n
maximized Ang II blockade had proteinuria >5 g per 24 h received two i.v. infusions of !po,Z&
rituximab at a dose of 1000 mg on days 1 and 15. To minimize infusion reactions, #w L(<nE
patients were premedicated with acetaminophen (1000 mg) and diphenhydramine n`=S&oKH
hydrochloride (50 mg) orally. In addition, methylprednisolone (100 mg, i.v.) was given <uIPv
Zsx
prior to the first rituximab infusion. B-cell depletion was defined as CD19+ count sA6Hk
B.
<5 cells per l at any time and B-cell recovery was defined as CD19+ cell count *4~7p4[
>15 cells per l. Patients treated with rituximab, who at month 6 had proteinuria >3 g per q[/g3D\G
24 h and in whom CD19+ B-cell counts had increased to >15 cells per l, received a ;7Qe m&
second course of rituximab treatment following the same protocol described above. yHM29fEZk
Follow-up `T[yyOL/
In all patients, clinical and laboratory parameters including complete blood counts, Mxz
X@GBX
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum qb nlD\
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry toF6 Z
and at months 1, 3, 6, 9, and 12. Creatinine clearance and protein and creatinine excretion AkjoD7.*
in the urine were assessed by performing two consecutive 24-h urine collections at each L,_U co
time point. Data were considered accurate when urinary creatinine excretion was P9\y~W
consistent with a complete 24 h collection. The mean of the two measurements was A f?&VD4K
considered for the analysis. The presence of HACAs was evaluated at baseline and at 9F,XjPK=
months 3, 6, 9, and 12. ;$a|4_U$m
Method / Approach / Study/ Technique *y"|/_
*
A discussion is presented of a problem-solving system s5 Fn("h]n
To improve the efficiency of the method, the following approach may be applied. 9LQy0Gx
In order to an investigation was made to find the causes of the ?fs#K;w
Although large collections of rules and equations have been complied, none are generally @Ng q+uXm
accepted suj}A
20 0g#?'sD
This approach will be explained and discussed thoroughly in the body of the report. $AFiPH9
This can be accomplished by "2"*3R<Y
This algorithm to compute the total cost can be described step by step as follows: `0ju=FP'u5
The above preliminary analysis has provided important information DA4!-\bt@
Various methods have been proposed for selecting an optimum... CQY/q@7
These concepts have been applied to #iGz&S3iN$
On the basis of the concept mentioned above, nuv$B >
This can be achieved by %@M/)"k
In addition, tissues were stained for infiltrating lymphocytes (CD20 and CD3), and the
K[TMTn
amount of interstitial fibrosis was quantified by histomorphometry. Formalin-fixed, M/sqOhg
paraffin-embedded sections were cut onto coated glass slides. Following heat-induced Hp> J,m(*
antigen retrieval, sections were incubated at 20°C overnight with either anti-CD20 f<'&_*7,|t
primary antibody or anti-CD3 primary antibody, both at 1:1000 dilution (Dako, Canada &(^>}&XS.<
Inc, Mississauga, Ontario, Canada). After rinsing all sections, pretreatment with 3% tT#Q`c
B
hydrogen peroxide was performed to prevent endogenous peroxidase activation. Sections .y[K =p3
were incubated with a secondary rabbit anti-mouse antibody linked with avidin–biotin 7iJk0L$]x
complex. Sections were counterstained with hematoxylin and examined by light s*#|EdD6@
microscopy. J2[QHr&tn
The HACA assay is a proprietary bridging enzyme-linked immunosorbent assay UZX)1?U
performed at Genentech Inc. that measures the antibody response to rituximab in human >%3c 1
serum samples. gS|xicq!
In all patients, clinical and laboratory parameters including complete blood counts, =<_5gR
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum *wF:Q;_<z
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry &s;%(c04A
and at months 1, 3, 6, 9, and 12. 9FLn7Y
This fact suggests that a new concept f8^58]wx0
This was accomplished by taking ... /#se>4]
The preparatory stage is very time consuming process. ?-84_i
Test are performed for validity, completeness, and compatibility 1=a}{)0h
There is little hope of achieving successful ... 4vQHr!$Ep
There has been an increasing awareness of the potential of using most ..so far made have Mqtp}<*@-
not taken this approach, with the exception of fDqXM;a"
Only a few studies can be found. D5m\u$~V
It is a very tedious process to go through
ck~xj0
It is only when .. has been completed that .. may be effected wqOhJYc
The entire interpretation process is conducted in one's head. /a|NGh%
These approaches are sometimes very tedious. RM+E
Several techniques can be used /nP=E
A polynomial parametric model can be written as [the following]/[follows]: r*wKYb
A xx model is constructed/formulated using xx. R*0F)M
A xx model represents an xx by its xx. W8NA.
A process decision model captures the logic essential to $
bosGG
From the equation above, xx is equal to the summation of xx times the ... vrv*k
21 mD)O\.uA
The validity of a xx model can be checked using Euler's formula. EvmmQ
Given a model, one can mathematically determine whether ... or ... b]#d04]
Equations for xx need to be derived and implemented in the system. 1[`<JCFClc
A number of heuristic rules have been developed for %Ab_PAw
Optimum .. techniques can be made more reliable by ... so that gu[dw3L
An algorithm based on the characteristic ... is used to determine & }}WP:U
Euler's formula states the following: k&s7-yY
The completed model should agree with the formula. F:S>\wG,
For manufacturing purposes, a detailed and precise model of the object is necessary <SZO-
-+lB
Engineering design models are very well defined; therefore, iBC>w+t14
To keep the domain narrow enough to be implementable, yet wide enough to be useful. f94jMzH9z
Point of View ld}$Tsy0
from an implementation standpoint, ]GPJ(+5
From the point of view of this application, zn1Rou]6
From this point of view, Zadeh suggested an inference rule named xxx (CRI for short). ;;hyjFGq%
Information is the meaningful interpretation and correlation of some aggregation of data A=d$ir
K[
in order to allow one to make decisions. YQdX>k
From a practical point of view, the computational aspects of an FLC require a 2RUR=%C
simplification of the fuzzy control algorithm. etMh=/NFV
The use of a hammer to insert screws, although partly effective, tends to distort, destroy, DE\bYxJ
and generally defeat the purpose of using a screw [Kusiak AI Implications for CIM T)
tZU?
p.129] !aw#',r8m
Statistical analysis o?mXxL)
Data were analyzed by one-way analysis of variance comparing the three conditions 0$L0fhw.
(sham operation, ischemic AKI, and bilateral nephrectomy) at each time point. If Iei7!KLW
significant F-statistic from analysis of variance existed, this test was followed by Dunnett ?t0zsq
post hoc multiple comparison procedure with sham operation as the control group. For all }#HTO:r
other comparisons, Student's t-test was used. A P-value of <0.05 was considered as Z\
=04[
statistically significant. SAG`^t
Values are expressed as means s.e.m. and significance was evaluated by Mann–Whitney >[H&k8\7n
U test using GraphPad Prism, version 4.0 software (GraphPad Software Inc., San Diego, B(\r+" PB
CA, USA). Go\} A:|s
All values are expressed as means s.d. Statistical significance (defined as P<0.05) was g7 Md
evaluated using analysis of variance and Bonferroni t-tests, and the two-tailed Pearson's
q]N?@l]
test, where appropriate. rI<nUy P?
Data are expressed as mean s.d., median and interquartile range, or frequencies, as uqa4&2(I=j
appropriate. Variables who deviated from the normal distribution (positively skewed) vNz;#Je
were log-transformed (log10) before the correlation study. S"P9Nf?9
Data are represented as means s.e.m. Student's t-test and multiple comparisons with t-test NDRW
post hoc analysis of variance were used as indicated below, for the comparison of Z\|u
9
DO
22 [Y22Wi
morphological, immunohistological and functional parameters. Statistical significance %wt2F-u
was set at P<0.05. DIG0:)4R.
The primary efficacy parameter was defined as change in urinary protein excretion from >P\h,1
baseline (week 0) to 12 months after treatment. The 12-month changes were tested 6wb^*dD92
against zero using the paired t-test. Secondary end points included 6-month changes in CJ9cCtA
protein; the number with PR or CR at 6 or 12 months; and changes in glomerular c$Xe.:QY
filtration rate (GFR), serum albumin, and lipid profiles. Study sample size was based on HQ^9[HN.
the desire to have 80% power to detect a drop in urinary protein of at least 2.0 g day-1. N
>d|A]zH
Assuming a two-sided hypothesis test performed at a significance level of 0.05 and an s.d. Jo{zy
of urinary protein change of 2.5 g, it was determined that 15 patients were required.2 kqVg2#<@M
Definition of remission status is according to the criteria established by Cattran et al.30 M`0(!Q}
CR was defined as proteinuria <0.3 g per 24 h, PR as proteinuria 3 g per 24 h, and a Qm.z@DwFM{
>50% reduction in peak proteinuria and non-response as <50% reduction in peak l^k+E-w\
proteinuria. Any patient reaching a CR or PR was considered a treatment success 3<c*v/L{C\
The statistical significance of differences for the mean values of cytokine concentration b/}'Vf[
and T cell proliferation was determined with Student's t-test. Differences with a P value `[WyHO|8
of less than 0.05 were considered significant. r_L
u~y|
Statistical significance was determined by Student's t-test. Data with a P value of less g-."sniP$g
than 0.05 were considered significant. |]^l^e6m
In vitro and in vivo experiments were analyzed by the two-tailed Student's t-test, with P SK's!m:r=
values less than 0.05 considered statistically significant. 5B%w]n
The significance of differences among groups was determined by Student's t-test and +Dv 7:x7
one-way analysis of variance (SigmaStat software; Jandel). #`/bQ~s
Comparisons ,1,&b_
As well, the pros and cons of these representations from a process planning point of view rB,ldy,f
will be discussed. 8+9\7*
The method of using xx to implement xx described by Zadeh (1973) appeared more JlUb0{8PE
suitable WhH!U0
As discussed [in the previous section]/[preciously], /w0sj`;"
Relation Wu ,S\!
We can not invert F' directly because it defines a many-to-one mapping. C#0brCQq3
The relationships appear very complicate F'1k<V?
Lifting tasks involve complex and imprecise relationship between the task variables and ^;+[8:Kb
the human operator's characteristics. 0*/kGvw`i
These methods are based on the relationship between ... and ... /
$_M@>
The fundamental concept of a fuzzy rating language is that we can establish a relationship h(nj,X+
among terms such as high, medium, and low, and then modify these relationships. =5?.'XMk
This article will thus mention the latter as well as the former. F_
81l<
The former two bear a close relation to a fuzzy Cartesian product. yplG18
Importance Ps3~{zH`
) Kk=LXmL2
23 QjJfE<h
The emphasis is on an implementation of a general approach to rule based decision JGis
" e
making. tb,9a!?
Consideration / Attention >'b=YlUL
Careful evaluation is necessary to ensure lI6W$V\,
Such a formulation does not change further considerations. :b@igZ<
Considerable attention has been paid to L5f$TLw
h;
Attention should be paid to an important finding of this investigation. 2 x32U
MD
Caution should be exercised in this process to avoid ... @^&7$#jq%
Primary consideration is given to ... components, though others can be accommodated D"^'.DL@wG
After ... has been defined by ..., a carefully analysis is carried out/performed to determine Ac`;st%l.
A number of factors such as ...need to be taken into consideration before making the
X{\jK]O
appropriate decision. (` c
G
It should be noted that 3{ "O,h
It is important to point out that ... +qy
6d7^
These considerations have heightened interest in the possibility of providing ... cG_Vc[
We should stress the fundamental importance of the xx ?lKhzH.T
Results. *yez:
qnx
Advantages / Disadvantage 5k$vlC#[H
One of the major advantages of this new measure of xx is that it can be applied to the HI
D([Wk
experimental study of zK;XFN#U^
One advantage of using a .. is the ease of preparing it. E!mmLVa9
It has a very fast decision making process D*2\{W/
All the algorithms involve mostly logical operations. OfZN|S+~W
It can be easily and without additional cost implemented in a microprocessor;based ^sClz*%?
environment. *y]+dK&-
It can reduce the waste of designing from scratch. fY W|p<Q0
The advantages of using a xx to represent xx are the following:
$NG++N
However, xx is not without its shortcomings. Or+p%K}-7
In most cases, the xxx shows an improvement over the existing xxx. eUBrzoCO
Compared to the existing xx, the impacts of the xx are generally reduced by 5% to 9%. VR0=SE
The "best case" results shows a savings of 6% to 9%. m9":{JI.w
Most of the existing works based on xx approach can only recognize a xx . L%h/OD
Most of the above methods are computational expansive and limited to xx. l=
={pb
Some other advantages of xx are the following: K5 KyG
The problem is the limitation of this method to a limited domain of parts. *hY2.t; X
It proved limited in application because it demanded precision in system modeling that wCKj7y[
was impossible in practice. I 3$dVls}
There are advantages to be gained in the structuring of costs and benefits, the use of xx, !8Rw O%c(
The disadvantages of this method are also disadvantages of conventional xx approaches. r4XH =
This combines the best features of both techniques A\sI<WrH
24 EqB)sK/3
Hopefully, this tool can be as the reference framework of for developing a xx platform, Ud(`V:d
and helping the administration, marketing, and knowledge management activities in kxB.,
'
virtual communities. pPro }@@
Results (p)!Mq
"^
An improvement on the result shown above can be made by based on the data provided c-4STPNQi
Recent work has identified dA~
3>f*b_
Time-dependent changes of }"s;\?a
Cyclosporine-induced cell death is triggered by a non-classical phosphoinositide 3-kinase lCT{v@pp
and does not require ERK activation. L,_.$1d
Much of the current work on j;~%
lg=)
Discussion of these theories is beyond the scope of this review Kq6jw/T
Based on the information contained in this chI.{Rj
The result can be categorized into nine classes HL K@xKD<
The results are illustrated by an example /F/;G*n
The experimental results for each xx time are reported in Table 2. 3R$Z[D-
From the results obtained so far, it seem that tln37vq
Because of the inaccuracy of the ..., a conclusion cannot be drawn as r?$?;%|C
Although much effort has been made to., this reality is far from completion. N@T.T=r
The results indicate that the total benefits are higher than the total costs. pW\'ZRj
Their results may then serve as guidelines for lower level models, less fuzzy and more a*dQ
_
detailed. io2)1cE&f
Conclusion u'~;Y.@i'
From the discussion, one may conclude that ... ~p1EF;4 #
The first indication that Zo-$z8
In summary, we have shown that 7;:#;YSha
These results suggested that .d<W`%[
Our findings have shown that %r{3wH#D@
Our observations have provided wucV_p.E
This study reveals following five main findings: LaFZ?7@|}
To our knowledge, this is the first immunohistochemical report of both PIM and HIFs in
-T@`hk`
the diabetic kidney. ?i\;:<e4
As mentioned above, fT\:V5-
Moreover, we demonstrate that, zt24qTKL
A number of studies have elaborated a body of knowledge about 1Nn@L2b 2
There are findings to indicate that q6#<[ 4?
On a descriptive level, there is agreement that for ;zTuKex~
One issue that became particularly evident from our study is +\
_{x/u1
Our data are in agreement with the findings and models presented by previous K-vWa2
publications, in particular those of Jones et al R{Z-m2La
According to our observations, ,V]
]:eR
…was not evident from our data F'PQqb {
25 M6|I6M
<
It could therefore be concluded that zR6siAV9
our data illustrate that klOp ^w
All these different aspects, as listed above, lead to ]DG?R68DQ
In fact, it has already been speculated that NLd``=&
It should be furthermore mentioned in this context that GOy%^:Xd
To identify and verify the essential factors triggering developmental renin expression, it ITJ
q
is of interest now to study the development of intrarenal renin expression in mice with o5o^TW{
defined gene mutations, using the results of this study as a reference system. oP?YA-#nc
In conclusion, using several approaches, we demonstrated that H[Q_hY[>V
But we have not provided formal proof yet that iTo k[uJ}
In summary, our results suggest that A1Y7;-D
In conclusion, we identify &NZfJs
A better understanding of such regulatory systems may yield novel therapeutic
cS.i
approaches to glomerular diseases. 4\5uY
Preliminary results of short-term studies (2 weeks) indicate that U!5)5c}G
However, we believe that `<" m%>
However, despite our extensive measurements of these parameters, we found no i7[uLdQ
relationship between response and initial proteinuria, time to B-cell recovery, the hPhZUL%
development of antibodies to rituximab, B-cell count in the kidney tissue, nor the degree .3 pbuU
of tubular interstitial damage present. A clear conclusion for efficacy, however, cannot be vC;]jJb:
obtained from an uncontrolled study, and definitive claims of efficacy should be reserved S*)1|~pRvQ
for rigorous, prospective, controlled, and randomized trials with the use of rituximab and ;(6g\'m
that will also look for factors that may determine its efficacy in IMN. "$HbK
@]!h
It has become increasingly clear that ^,Ydr~|T
Form the above discussion, the conclusion can be reached that _1Z=q.sC
The conclusions drawn are also valid Mq%,lJA\
In conclusion to this, it becomes obvious that the problem of xx lies not only in... s$h]
G[x
We have attempted to introduce some concepts associated with a theory of 3#kitmV
Considerable more work, hopefully, will be done in this area \Xrw"\")j
Employing the compositional rule of inference, the assessment of the xx compatibility in ;+ "f
achieving prescribed xx projectiles in any level of the hierarchy is made possible. pj|pcv^
This paper has presented a theoretical and experimental study of the xx process and xx 5 +9Ze9
concept. f5RE9%.#~
The experimental research results will hopefully serve as useful feedback information for e59dVFug.U
improvements for xx work. Q'apG)0I
The scope of this contribution was to introduce a xx method. Z5G!ct:W
This has a tremendous impact on our understanding of }$su4A@0
Significant progress has been made in advancing RNAi therapeutics in a remarkably 1pp -=$k
short period of time J a,d3K
To date, little is known about… lEL78l.
Irrespective of the mechanisms and the molecules –which are still largely unknown – JEn3`B!*
involved in MSC functions, current data suggest that <T?oKOD ]
To better understand &`|:L(+
26 ^o?S M^
Future Research (常用于conclustion 的结尾) eZhPu'id\s
Thus, first extension of the approach could be, qgt[ ~i*
Further studies are needed to determine the importance of `\>.h
Some improvements to the scheduling aspect of the model may be brought through z1OFc
qm
additional levels in the hierarchy for more detailed representation of the scheduling @=S}=cl
activity. ]T!
}XXK
It also unveiled new aspects of the role played by thyroid hormone in response to injury aAvsb$
and tissue repair. Zx }&c |Q
In the near future, the ongoing clinical trials with siRNAs for macular degeneration and $>M<j
RSV may reveal the exciting potential of RNAi therapeutics as the next major class of =`n]/L"Q
drug molecules. Vv
bFp
In the next few years it should become clear whether dGfVZDsr]
It should be an exciting time for researchers seeking to harness this powerful endogenous _:n b&B
pathway to treat human disease. vh!v
MB}}
Acknowledgement SZ![%)83
The authors are grateful to the insightful work provided by Mr. John W. Harney and all ^ OJyN,A
of the fellows and students that contributed with some of the studies discussed in this ;Lw{XqT
article ^_I} x)i*@
I would like to express my gratitude to all those who gave me the possibility to complete kpwt]]e*
this thesis. *x~xWg9^
I want to thank the Department of Transport of the former Interim Government of ;P &y,:<m:
Namibia for giving me permission to commence this thesis in the first instance, to do the $!vxVs9n
necessary research work and to use departmental data :vx$vZb
This work was supported by NIH grants AI058695 and AI056900. We thank members of ,iNv'
the laboratory and our collaborators for many useful suggestions. vP%tk s+.
We thank J. Maraganore and B. Greene for critical reading of the manuscript, M. Mj>}zbpk/
Duckman for graphics assistance and A. Capobianco for word processing assistance. .Hhh i
Author contributions J;t 7&Zpe
All authors were involved in experimental planning and data analysis and contributed to 9Ywpej*+
manuscript preparation. 0oA{Jix
Tables and Figures [./6At&|
Figure 7-1 sketches these relationships. #BVtL :x@
The graphical representation of these functions is shown in Figure 1. P;ZU-G4@
The xx may be depicted as in Figure 1. Y&H}xn
Figure x shows the schematic diagram of the h/eR
Figure 1 though 2 provide a ... that wCs^J48=
the architecture of this expert system for .... is illustrated in Figure 2. -Lu)'+
Figure 2 gives the outline of an ... system c'6g*%2k
Table shows the }uHrto3M
27 W+wA_s2&D
Time-dependent changes of WFR?fDtE
as shown in Table 1 and 2 S0h'50WteJ
This concept is illustrated in Figure 2 75nNh~?)\
At the top of Table xx are shown two blocks of data. 4!Z5og1kn
Each table or matrix has constructs xx through xx as row;headings, RpG+>"1]
xx through xx as column;headings. j hYToMq
Time course of calculated renin immunoreactive tissue volumes during development of mE O\r|A
the mouse kidney. Data are single values per time point. They were derived from the Wc3kO'J
kidneys shown in Figure 1. 0tISXu-
Yellow color indicates lPH%Do>K
PCT3 cells were pretreated with 50 M PD98059 or 10 M U0126 for 30 min and then !f>d_RG
cells were stimulated with 25 g ml-1 CsA for 6 h. ERK and PKB activation were xX3'bsN
analyzed by western blot using phospho-specific antibodies. 4p%A8%/q
ERK and PKB activation were measured by western blot using phospho-specific 7e7 M@8+4
antibodies 4v
p
The data are the means of three different experiments taking the number of untreated cells
d\e7,"L*Q
(vehicle alone) as 100%. &<zd.~N"
ERK and PKB activation were then analyzed by western blot with the corresponding ]v[|B
phospho-specific antibodies. To ensure equal amount of protein in each lane, western }:: S0l
blots against total protein were performed for ERK and PKB. _X~87
The means of optical density (OD) values of the bands detected in western blots of three :^UFiUzrE
different experiments are plotted, taking the maximal value of phosphorylation of ERK ntNI]~z&
and PKB as 100%. (SSRY 9
CT3 cells were treated with 25 g ml-1 CsA or vehicle alone for 24 h and then fixed with .xf<=ep
paraformaldehyde and stained with Hoescht as indicated in Materials and Methods. Then dEBcfya
cells were visualized by phase-contrast microscopy (upper panel) and fluorescence "CiTa>x
microscopy (lower panel). ^Rm
The data are the means of three experiments performed on different days taking the cj11S>D
number of cells at the onset of the experiment as 100%. ^
9!!;)
Rats were killed at 4 h and at days 1, 2, 4, 7, 9, 14, 21, and 28 after disease induction ;?C#IU
(n=9 each). 3D?sL!W
*P<0.05 versus non-nephritic rats. 54zl
nM$
The expression of the CCN3 protein was detected by western blot analysis (n=4 each, PC, Z{'i
F
positive control, M, molecular weight markers). p~LrPWHSTP
Original magnifications: 200 in A, E, G, H; 600 in B, C, D, F. 26CS6(sn
Data are means s.d. of four independent experiments. zbJT&@z
*P<0.05 versus 0.5 h. .k,YlFvj
PDGF-BB and -DD, but not PDGF-AA and -CC, induce a downregulation of CCN3 |Ph3#^rM?
mRNA as determined by real-time RT-PCR and normalized to B=|cS;bM$3
glyceraldehyde-3-phosphate dehydrogenase mRNA. N
Hlk|Y#6b
Cell number was counted in duplicate using a Malassez hemocytometer XrZ
*1V
Data are means s.d. of four independent experiments. *P<0.05 versus cells treated with ,AT[@
MCDB media for 24 h, #P<0.05. >M&3Y
XC
28 3Gkv4,w<
The curves are adjusted for history of coronary artery disease, CRP, and progression to _UuC,Pl3
ESRD. :%IoM E
Both models are independent of age, gender, baseline GFR and proteinuria, and other @B}aN@!/
clinical characteristics and traditional and nontraditional cardiovascular risk factors W^"AU;^V56
Correlation between serum bone-specific alkaline phosphatase (bAP) and serum PTH Y".RPiTL
(n=99, P<0.0001, r2=0.190) fDLG>rXPT
Concentration of serum FGF23 concentration before starting and at the end of the 4-h itMg|%B%
hemodialysis procedure in 23 patients, expressed as log10 values (n=23, P<0.0001, 1@$Ko5
Student's paired t-test). bJD;>"*
Immunohistochemistry for the hypoxia marker pimonidazole (PIM) and HIF-2 ; K<>sOWZ'S
arrow=endothelial cell and CD=collecting duct Dih3}X&jn$
Magnification: 1000. ^b
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