18 R5eB,FN
Jurkat cells cultured in calcium-free RPMI-1640 medium (Gibco BRL; number kr(<Y|
22300-107) containing calcium-free 10% FBS were triggered by anti-Fas IgM. The <i'u96
treated cells were harvested at the indicated time points and incubated with Fluo-3-AM at ES\Q5)t/fo
a final concentration of 1 micromolar for 30 min at 37°C (Scoltock et al., 2000). The )6>|bmpU
labeled cells (50,000 cells per treatment) were then analyzed by exciting the cells at 488 cvUut^CdK
nm and examining the fluorescence emission of Fluo 3 at 530 nm with a FACS Scan, @ 9 {%Kn
(Becton Dickinson). A one micromolar concentration of LPA was used as a positive [9O~$! <%
control for Ca2+ induction. The data thus obtained was analyzed with the software Win QG|GXp_q`
MDI 2.8 and represented as contour plots.The effect of chelating intracellular calcium on VL7S7pb_
translocation of annexin I was studied by culturing Jurkat cells in the presence of 10 G|-\T(&J
micromolar BAPTA-AM, with or without the addition of anti-Fas IgM. Cells were PDt<lJU+X
harvested and fractionated as detailed above, and the S-100 fractions were assessed by D*cyFAF
immunoblotting for presence or absence of annexin I. qggk:cN1
Mouse bone marrow transduction and transplantation. c',:@2R
Retrovirus-mediated transduction of mouse bone marrow cells was done by published IE)"rTI)b
methods49. Prestimulated low-density bone marrow cells were infected with high-titer ~zxwg+:QO
retrovirus supernatant on fibronectin-coated plates. Retrovirus supernatant was generated x_<bK$OU
in the phoenix-gp cells with a mouse stem cell virus–based retroviral vector coexpressing [
vWkAJ'K
EGFP and HA–RhoH as described50. EGFP+ sorted cells were transplanted by I/H
V;g:#
intravenous injection into the sublethally irradiated (300 rads with a 137Cs irradiator) R3
j#WgltP
Rag2-/- recipient mice. At 9 weeks after transplantation, thymus, peripheral blood, bone {^cF(7p
marrow, spleen and lymph nodes from each recipient mouse were collected for analysis A~S
L5h
of EGFP+ chimerism and hematopoietic lineage by flow cytometry. Expression of <!~1{`n%9J
HA–RhoH and HA–RhoHF73F83 in EGFP+ sorted thymocytes of recipient mice was $_RWd#Q(
confirmed by immunoblot analysis. vm\wO._
Determination of renal morphology ACigeK^C}E
Kidney slices were postfixed in buffered 2% OsO4, dehydrated, and embedded in an 9zZ5Lr^21
Araldite-EM bed 812 mixture. Large sections were cut perpendicular to the renal capsule, StU 4{
containing cortex, and medulla. Thin (1 m) sections were analyzed in a blinded manner *N-;V|{
for morphologic alterations, as previously detailed 7m:|u*ij2~
Patient population RpO@pd m
Patients included in the study met the following criteria: (1) biopsy-proven IMN; (2) /l,+oG%\
creatinine clearance 30 ml per min per 1.73 m2; and (3) persistent proteinuria >5 g per sI&i{D
24 h despite treatment with an HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A) z\fW )/
reductase inhibitor, an ACEi, and/or ARB at maximal tolerated dose for at least 4 months. khU6*`lQ
The Mayo Institutional Review Board and the Research Ethical Board, University Health M{?zvq?d
Network, University of Toronto approved the study protocol. All patients gave written ;rB6u_5"I.
informed consent. Patients who had been on treatment with prednisone, cyclosporine, or Rx6l|'e
19 &?.n2+T+
=
mycophenolic mofetil within the last 4 months or alkylating agents within the last 6 "]V
DY)
months were not included in the study. Patients with active infection, diabetes, or a ^WE4*.(
secondary cause of MN (for example, hepatitis B, systemic lupus erythematosus (SLE), #D+7TWDwNt
medications, malignancies) were also excluded. Y^52~[w~
Treatment S6v!GQ
At enrollment, a low-sodium (<4 g day-1) and low-protein (0.8 g per kg per day of P$Z}
high-quality protein) diet was recommended and patients were encouraged to maintain OT%0{2c"]
the same diet throughout the duration of the study. All patients received a similar 9-0<*)"b>
conservative treatment regimen that included loop diuretics to control edema, an !^m%O0DT
HMG-CoA reductase inhibitor, and an ACEi combined with an ARB if tolerated. d1v<DU>M
-Blockers and non-dihydropyridine calcium channel blockers, in that order, were added 9O;cJ)tXY
when required to control systolic blood pressures to <135 mm Hg in >75% of the GCDwWCxh
readings. Patients who after a minimum of 4 months of conservative therapy and qv[[Q[RK-5
maximized Ang II blockade had proteinuria >5 g per 24 h received two i.v. infusions of !p9)CjQ "
rituximab at a dose of 1000 mg on days 1 and 15. To minimize infusion reactions, veeI==]
patients were premedicated with acetaminophen (1000 mg) and diphenhydramine 33Jd!orXU
hydrochloride (50 mg) orally. In addition, methylprednisolone (100 mg, i.v.) was given ~9GOk;{~&
prior to the first rituximab infusion. B-cell depletion was defined as CD19+ count .XXW
|{
<5 cells per l at any time and B-cell recovery was defined as CD19+ cell count M=O Czgj
>15 cells per l. Patients treated with rituximab, who at month 6 had proteinuria >3 g per 3jjMY
24 h and in whom CD19+ B-cell counts had increased to >15 cells per l, received a 07V8;A<,
second course of rituximab treatment following the same protocol described above. '+j<n[JLC
Follow-up C|MQ
$~5:w
In all patients, clinical and laboratory parameters including complete blood counts, bLzs?eos
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum ~C-,G"zw&G
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry +TK3{5`!Ae
and at months 1, 3, 6, 9, and 12. Creatinine clearance and protein and creatinine excretion ~@T<gA9V
in the urine were assessed by performing two consecutive 24-h urine collections at each ~vH
k&r]|
time point. Data were considered accurate when urinary creatinine excretion was As~(7?]r
consistent with a complete 24 h collection. The mean of the two measurements was #
2RiLht
considered for the analysis. The presence of HACAs was evaluated at baseline and at !gF9k8\Yr$
months 3, 6, 9, and 12. F>zl9Vi<
Method / Approach / Study/ Technique G?AG:%H %
A discussion is presented of a problem-solving system |Os6V<u"
To improve the efficiency of the method, the following approach may be applied. $<nCXVqL,
In order to an investigation was made to find the causes of the W3r?7!~
Although large collections of rules and equations have been complied, none are generally .~>?*}
accepted G,f-.
20 4[yIOs
This approach will be explained and discussed thoroughly in the body of the report. &;E5[jO^D
This can be accomplished by
%ft &Q
This algorithm to compute the total cost can be described step by step as follows: ?YQPlv:<o.
The above preliminary analysis has provided important information %'
$o"
Various methods have been proposed for selecting an optimum... fpWg R4__
These concepts have been applied to \25/$
Ae}c
On the basis of the concept mentioned above, :O-iykXyI
This can be achieved by |TsE-t*E}
In addition, tissues were stained for infiltrating lymphocytes (CD20 and CD3), and the M;w?[yEZ
amount of interstitial fibrosis was quantified by histomorphometry. Formalin-fixed, #3$\Iu
paraffin-embedded sections were cut onto coated glass slides. Following heat-induced oVvc?P
antigen retrieval, sections were incubated at 20°C overnight with either anti-CD20 5| B(\wqG
primary antibody or anti-CD3 primary antibody, both at 1:1000 dilution (Dako, Canada m<rhIq
Inc, Mississauga, Ontario, Canada). After rinsing all sections, pretreatment with 3% Yd
EptAI
hydrogen peroxide was performed to prevent endogenous peroxidase activation. Sections K+*Q@R D
were incubated with a secondary rabbit anti-mouse antibody linked with avidin–biotin /./"x~@
complex. Sections were counterstained with hematoxylin and examined by light z~*g ~RKS!
microscopy. ~y HU^5D
The HACA assay is a proprietary bridging enzyme-linked immunosorbent assay r]@0eb
performed at Genentech Inc. that measures the antibody response to rituximab in human *NFg;<:j
serum samples. 7hP<f}xL
In all patients, clinical and laboratory parameters including complete blood counts, H}hFFI)#Oo
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum ldha|s.*
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry mw$r$C{
and at months 1, 3, 6, 9, and 12. az0<5Bq)
This fact suggests that a new concept /1N6X.Zb
This was accomplished by taking ... 5r~#0Zf*
The preparatory stage is very time consuming process.
3E3U /K
Test are performed for validity, completeness, and compatibility ;~fT,7qBah
There is little hope of achieving successful ... Hnknly
There has been an increasing awareness of the potential of using most ..so far made have >r`b_K
not taken this approach, with the exception of SIO&rrT.
Only a few studies can be found. +#U|skl
It is a very tedious process to go through qm/Q65>E
It is only when .. has been completed that .. may be effected =_$Qtq+h
The entire interpretation process is conducted in one's head. Q!-
0xlx
These approaches are sometimes very tedious. 3 LDS
Z1f
Several techniques can be used &'cL%.
A polynomial parametric model can be written as [the following]/[follows]: valtev0<
A xx model is constructed/formulated using xx. Z^ }mp@j>
A xx model represents an xx by its xx. Lg8nj< TF
A process decision model captures the logic essential to
'iLpE7
From the equation above, xx is equal to the summation of xx times the ... ~wg:!VWA)
21 BA\/YW @
The validity of a xx model can be checked using Euler's formula. .MO\uh0N
Given a model, one can mathematically determine whether ... or ... "KcA
Equations for xx need to be derived and implemented in the system. z/i+EE
A number of heuristic rules have been developed for r0p w_j
Optimum .. techniques can be made more reliable by ... so that $kR N
h6
An algorithm based on the characteristic ... is used to determine oIUy -|
Euler's formula states the following: rG,5[/l
The completed model should agree with the formula. UK9@oCIB
For manufacturing purposes, a detailed and precise model of the object is necessary ^C2\`jLMY
Engineering design models are very well defined; therefore, a0r"N[&
To keep the domain narrow enough to be implementable, yet wide enough to be useful. hiNEJ_f
Point of View ^vG8#A}]
from an implementation standpoint, >0Q|nCx
From the point of view of this application, wAnb
Di{W
From this point of view, Zadeh suggested an inference rule named xxx (CRI for short). zYl#
4O`=c
Information is the meaningful interpretation and correlation of some aggregation of data sz9L8f2
in order to allow one to make decisions. OY?y ^45y
From a practical point of view, the computational aspects of an FLC require a !^Q.VYY
simplification of the fuzzy control algorithm. iNMx"F0r
The use of a hammer to insert screws, although partly effective, tends to distort, destroy, i<pk6rO1
and generally defeat the purpose of using a screw [Kusiak AI Implications for CIM 1sYEZO;
p.129] 'Ei;^Y 1e
Statistical analysis YM5fyv?
Data were analyzed by one-way analysis of variance comparing the three conditions Bn>8&
w/P
(sham operation, ischemic AKI, and bilateral nephrectomy) at each time point. If *L!!]Q2c
significant F-statistic from analysis of variance existed, this test was followed by Dunnett g2unV[()_
post hoc multiple comparison procedure with sham operation as the control group. For all HF3f)}l$
other comparisons, Student's t-test was used. A P-value of <0.05 was considered as \>Efd
statistically significant.
D %JlbH8
Values are expressed as means s.e.m. and significance was evaluated by Mann–Whitney ^iQn'++Q
U test using GraphPad Prism, version 4.0 software (GraphPad Software Inc., San Diego, $+7`Dy!
CA, USA). $8a(veXd
All values are expressed as means s.d. Statistical significance (defined as P<0.05) was 37;
$-cFE
evaluated using analysis of variance and Bonferroni t-tests, and the two-tailed Pearson's
S4=R^];l
test, where appropriate. hR.@b*q?R
Data are expressed as mean s.d., median and interquartile range, or frequencies, as Z}-Vf$O~
appropriate. Variables who deviated from the normal distribution (positively skewed) }x&N^Ky3c
were log-transformed (log10) before the correlation study. n`4K4y%Dy}
Data are represented as means s.e.m. Student's t-test and multiple comparisons with t-test ^ "\R\COQ
post hoc analysis of variance were used as indicated below, for the comparison of /Wf^hA
22 `%j~|i)4
morphological, immunohistological and functional parameters. Statistical significance v(pmIb{
was set at P<0.05. Y[ N^p#t{
The primary efficacy parameter was defined as change in urinary protein excretion from _!|/
;Nk
baseline (week 0) to 12 months after treatment. The 12-month changes were tested >"Q@bQ:e
against zero using the paired t-test. Secondary end points included 6-month changes in 7==f\%,
protein; the number with PR or CR at 6 or 12 months; and changes in glomerular ~xoF6CF
filtration rate (GFR), serum albumin, and lipid profiles. Study sample size was based on m XXt'_"
the desire to have 80% power to detect a drop in urinary protein of at least 2.0 g day-1. mq%<6/YU
Assuming a two-sided hypothesis test performed at a significance level of 0.05 and an s.d. T,$WlK
Wj
of urinary protein change of 2.5 g, it was determined that 15 patients were required.2 [b$4Shx
Definition of remission status is according to the criteria established by Cattran et al.30 xz:J
CR was defined as proteinuria <0.3 g per 24 h, PR as proteinuria 3 g per 24 h, and a ~.!c~
fke
>50% reduction in peak proteinuria and non-response as <50% reduction in peak *cO sv
proteinuria. Any patient reaching a CR or PR was considered a treatment success %sRUh0AL
The statistical significance of differences for the mean values of cytokine concentration c,*9K/:
and T cell proliferation was determined with Student's t-test. Differences with a P value ]TaN{"
of less than 0.05 were considered significant. 7h)iu9j
Statistical significance was determined by Student's t-test. Data with a P value of less TB;o~>9U
than 0.05 were considered significant. "H
XYNS>
In vitro and in vivo experiments were analyzed by the two-tailed Student's t-test, with P }E_#k]#*
values less than 0.05 considered statistically significant. vip~'
The significance of differences among groups was determined by Student's t-test and 3su78e t}
one-way analysis of variance (SigmaStat software; Jandel). ti &J
Comparisons pc?>cs8
As well, the pros and cons of these representations from a process planning point of view qn,O40
/]
will be discussed. (os$B
The method of using xx to implement xx described by Zadeh (1973) appeared more {%#)5l)
suitable M,5"b+mX[~
As discussed [in the previous section]/[preciously], `~X!Ll
Relation 4l&g6YneX
We can not invert F' directly because it defines a many-to-one mapping. =LTmr1?
The relationships appear very complicate ,<:!NF9
Lifting tasks involve complex and imprecise relationship between the task variables and ;us%/kOR
the human operator's characteristics. _{|D
These methods are based on the relationship between ... and ... B:Z_9,gj-N
The fundamental concept of a fuzzy rating language is that we can establish a relationship C4}*)a
among terms such as high, medium, and low, and then modify these relationships. h!m_PgRSs
This article will thus mention the latter as well as the former. i>[_r,-\[
The former two bear a close relation to a fuzzy Cartesian product. Qeu\&%C!<
Importance H}LS??P
) DI;DECQl$
23 <x\7L2#p
The emphasis is on an implementation of a general approach to rule based decision zm,@]!wI
making. (Y^X0yA/
Consideration / Attention RaTH\>n
Careful evaluation is necessary to ensure R1Ye<R!Q
Such a formulation does not change further considerations. Va$Pi19 O
Considerable attention has been paid to L}m8AAkP[
Attention should be paid to an important finding of this investigation. 8?Z4-6!{V,
Caution should be exercised in this process to avoid ... rDdzxrKg{
Primary consideration is given to ... components, though others can be accommodated _
%%Z6x(
After ... has been defined by ..., a carefully analysis is carried out/performed to determine +ki{H}G21
A number of factors such as ...need to be taken into consideration before making the }KD;0t4
appropriate decision.
8L*GE
It should be noted that [Ei1~n)o
It is important to point out that ... GjB]KA^
These considerations have heightened interest in the possibility of providing ... it2 a
We should stress the fundamental importance of the xx `m.eM
Results. hAj1{pA,
Advantages / Disadvantage ""s
vDfy$
One of the major advantages of this new measure of xx is that it can be applied to the 1oKfy>i e
experimental study of $W;b{H=F
One advantage of using a .. is the ease of preparing it. *1@:'rJ
It has a very fast decision making process eh R{X7J
All the algorithms involve mostly logical operations.
~m*,mz
It can be easily and without additional cost implemented in a microprocessor;based IHC1G1KW=A
environment. ?GBkqQ
It can reduce the waste of designing from scratch. dC}`IR
The advantages of using a xx to represent xx are the following: LEEC W_:
However, xx is not without its shortcomings. (5^ZlOk3
In most cases, the xxx shows an improvement over the existing xxx. 2l5KJlfj>k
Compared to the existing xx, the impacts of the xx are generally reduced by 5% to 9%. nY $tp
The "best case" results shows a savings of 6% to 9%. UofTl
l)
Most of the existing works based on xx approach can only recognize a xx . n-QJ;37\
Most of the above methods are computational expansive and limited to xx. z<]bv7V
Some other advantages of xx are the following: ^/Sh=4=G
The problem is the limitation of this method to a limited domain of parts. sCl$f7"
It proved limited in application because it demanded precision in system modeling that YxJD _R
was impossible in practice. bzi|s5!'<
There are advantages to be gained in the structuring of costs and benefits, the use of xx, ;7Oi! BC
The disadvantages of this method are also disadvantages of conventional xx approaches. ji+{ :D
This combines the best features of both techniques fRTo.u
24 @;wzsh >o
Hopefully, this tool can be as the reference framework of for developing a xx platform, 5(&xNT-n8
and helping the administration, marketing, and knowledge management activities in H^1gy=kdj
virtual communities. /)uM[ dnai
Results 0"[`>K~7a8
An improvement on the result shown above can be made by based on the data provided c#xP91.m
Recent work has identified 5|$a =UIR
Time-dependent changes of rn.\tDeA
Cyclosporine-induced cell death is triggered by a non-classical phosphoinositide 3-kinase `0#H]=$2h
and does not require ERK activation. MEwdw3
Much of the current work on @kD8^,( oH
Discussion of these theories is beyond the scope of this review _|isa]u\z
Based on the information contained in this 3]E(mRX
The result can be categorized into nine classes >Cd9fJ&0gP
The results are illustrated by an example 1K|@h&@
The experimental results for each xx time are reported in Table 2. 8
|h9sn;P
From the results obtained so far, it seem that u}H$-$
jE
Because of the inaccuracy of the ..., a conclusion cannot be drawn as /c+)C"
Although much effort has been made to., this reality is far from completion. B94
&elu
The results indicate that the total benefits are higher than the total costs. M:ai<TZ]
Their results may then serve as guidelines for lower level models, less fuzzy and more k ~6-cx
detailed. ]@xL=%
Conclusion _
py2kjA6
From the discussion, one may conclude that ... <6N_at3
The first indication that ?p8k{N(1
In summary, we have shown that 2DMrMmLI
These results suggested that 5)lW
Our findings have shown that ehO:')XF
Our observations have provided &^ I+s^\=
This study reveals following five main findings: V;9 }7mw
To our knowledge, this is the first immunohistochemical report of both PIM and HIFs in
fCEz-TMW
the diabetic kidney. StQ@g
As mentioned above, ~y\:iL//E
Moreover, we demonstrate that, U|fTb0fB
A number of studies have elaborated a body of knowledge about <]#o*_aFP
There are findings to indicate that $@87?Ab
On a descriptive level, there is agreement that for q6[}ydV
One issue that became particularly evident from our study is t
9*=
Our data are in agreement with the findings and models presented by previous `facFt[\
publications, in particular those of Jones et al lIDl1Z@Z
According to our observations, >C2HC6O
3
…was not evident from our data )}|
mDN&P
25 nu 7lh6o=
It could therefore be concluded that T]2U fi.
our data illustrate that 2YIF=YWO},
All these different aspects, as listed above, lead to 6D*chvNA;
In fact, it has already been speculated that ^('cbl
It should be furthermore mentioned in this context that ^-L{/'[8M
To identify and verify the essential factors triggering developmental renin expression, it }1rvM4{/+f
is of interest now to study the development of intrarenal renin expression in mice with [#X|+M&u6
defined gene mutations, using the results of this study as a reference system. P
dM*5g4
In conclusion, using several approaches, we demonstrated that ] piM
/v\
But we have not provided formal proof yet that 8m7eaZ
In summary, our results suggest that zv9MHC
&
In conclusion, we identify f=oeF]=I"
A better understanding of such regulatory systems may yield novel therapeutic yam}x*O\xn
approaches to glomerular diseases. |J
?:91
Preliminary results of short-term studies (2 weeks) indicate that .WO/=#O
However, we believe that g =x"cs/[
However, despite our extensive measurements of these parameters, we found no V(I7*_ZFl
relationship between response and initial proteinuria, time to B-cell recovery, the SauHFl8?
development of antibodies to rituximab, B-cell count in the kidney tissue, nor the degree . z].:$J&
of tubular interstitial damage present. A clear conclusion for efficacy, however, cannot be 4a 4N
C
obtained from an uncontrolled study, and definitive claims of efficacy should be reserved #'g^Za
for rigorous, prospective, controlled, and randomized trials with the use of rituximab and {0fz9"|U
that will also look for factors that may determine its efficacy in IMN. wEK%T P4
It has become increasingly clear that aYBTrOd z
Form the above discussion, the conclusion can be reached that O$(c.(_$
The conclusions drawn are also valid ,M{Q}:$+4
In conclusion to this, it becomes obvious that the problem of xx lies not only in... @5GBuu^j
We have attempted to introduce some concepts associated with a theory of ~ ;XYwQ"
Considerable more work, hopefully, will be done in this area !3QRzkJX~
Employing the compositional rule of inference, the assessment of the xx compatibility in /Bm#`?(ia
achieving prescribed xx projectiles in any level of the hierarchy is made possible. Kc-4W6?$
This paper has presented a theoretical and experimental study of the xx process and xx kI<WvgoL
concept. To">DOt
The experimental research results will hopefully serve as useful feedback information for UIz:=DJ
improvements for xx work. $(XgKq&xWZ
The scope of this contribution was to introduce a xx method. yz8-&4YRNd
This has a tremendous impact on our understanding of ,y{0bq9*2
Significant progress has been made in advancing RNAi therapeutics in a remarkably k=ts&9\
short period of time ">4PePt.n
To date, little is known about… fqN75['n
Irrespective of the mechanisms and the molecules –which are still largely unknown – JxnuGkE0[#
involved in MSC functions, current data suggest that (Hl8U
To better understand Q%I#{+OT
26 \hrrPPD1z
Future Research (常用于conclustion 的结尾) {v,{x1
Thus, first extension of the approach could be, kq>GMUl~@
Further studies are needed to determine the importance of }'DC
Q
Some improvements to the scheduling aspect of the model may be brought through mF}c-
D
additional levels in the hierarchy for more detailed representation of the scheduling Ut]2` 8-
activity. 6$;L]<$W>
It also unveiled new aspects of the role played by thyroid hormone in response to injury Q
js2hj-$
and tissue repair. E2kW=6VO>|
In the near future, the ongoing clinical trials with siRNAs for macular degeneration and OI*ZVD)J
RSV may reveal the exciting potential of RNAi therapeutics as the next major class of 1;aF5~&
drug molecules. R!@|6=]iG
In the next few years it should become clear whether w2 /* `YO
It should be an exciting time for researchers seeking to harness this powerful endogenous |MR%{ZC^i
pathway to treat human disease. E2Us#a
Acknowledgement 4 #aqz9k
The authors are grateful to the insightful work provided by Mr. John W. Harney and all &S<?07Z
of the fellows and students that contributed with some of the studies discussed in this XO 0>t{G
article
CdDH1[J
I would like to express my gratitude to all those who gave me the possibility to complete K?9WY]Ot
this thesis. $U ~=.!_du
I want to thank the Department of Transport of the former Interim Government of PP!l
Namibia for giving me permission to commence this thesis in the first instance, to do the Tku/OG'
necessary research work and to use departmental data Yr5iZ~V$
This work was supported by NIH grants AI058695 and AI056900. We thank members of $<
A8gTJ
the laboratory and our collaborators for many useful suggestions. I0I_vu
We thank J. Maraganore and B. Greene for critical reading of the manuscript, M. Ok*VQKyDLH
Duckman for graphics assistance and A. Capobianco for word processing assistance. mgo'M
W\
Author contributions CFD*g\g<*
All authors were involved in experimental planning and data analysis and contributed to 4\&H?:c.
manuscript preparation. g4zT(,ZY
Tables and Figures mG>T`c|r3
Figure 7-1 sketches these relationships. 's =Q.s
The graphical representation of these functions is shown in Figure 1. {'h)
The xx may be depicted as in Figure 1. /uC+.B9k
Figure x shows the schematic diagram of the ;n3uV`\
Figure 1 though 2 provide a ... that &6`
the architecture of this expert system for .... is illustrated in Figure 2. T^KCB\\<
Figure 2 gives the outline of an ... system 0\V\qAk
Table shows the )3k)2X F
27 U,oD44
Time-dependent changes of [
P.@1mV
as shown in Table 1 and 2 _pY
This concept is illustrated in Figure 2 ~+nS)4(
At the top of Table xx are shown two blocks of data. Hi9 G^Q
Each table or matrix has constructs xx through xx as row;headings, p5lR-G
xx through xx as column;headings. Q.j-C}a
Time course of calculated renin immunoreactive tissue volumes during development of 1h#w"4
the mouse kidney. Data are single values per time point. They were derived from the ZrmnQ
kidneys shown in Figure 1. /AK*aRU^
Yellow color indicates 2&^,IIp
PCT3 cells were pretreated with 50 M PD98059 or 10 M U0126 for 30 min and then /.pa
??u
cells were stimulated with 25 g ml-1 CsA for 6 h. ERK and PKB activation were y}(_SU
analyzed by western blot using phospho-specific antibodies. `77;MGg*
ERK and PKB activation were measured by western blot using phospho-specific 8r7/IGFg
antibodies Y}V)4j
The data are the means of three different experiments taking the number of untreated cells a(Gk~vD;"
(vehicle alone) as 100%. J,dG4.ht
ERK and PKB activation were then analyzed by western blot with the corresponding |YG)NO
phospho-specific antibodies. To ensure equal amount of protein in each lane, western J8qu]{0I"
blots against total protein were performed for ERK and PKB. Y-}hNZn"{
The means of optical density (OD) values of the bands detected in western blots of three #C;#$|d
different experiments are plotted, taking the maximal value of phosphorylation of ERK zJB+C=]D7H
and PKB as 100%. ^L*VW
gi9
CT3 cells were treated with 25 g ml-1 CsA or vehicle alone for 24 h and then fixed with ~2, wI<Nz
paraformaldehyde and stained with Hoescht as indicated in Materials and Methods. Then 18[?
dV
cells were visualized by phase-contrast microscopy (upper panel) and fluorescence ql%]$`IV6
microscopy (lower panel). x| ~D(zo
The data are the means of three experiments performed on different days taking the uK"$=v6|
number of cells at the onset of the experiment as 100%. ,b2O^tJF#
Rats were killed at 4 h and at days 1, 2, 4, 7, 9, 14, 21, and 28 after disease induction o'= [<
(n=9 each). NO0[`jy(
*P<0.05 versus non-nephritic rats. 6`$,-(J=
The expression of the CCN3 protein was detected by western blot analysis (n=4 each, PC, {ra Esb-X
positive control, M, molecular weight markers). ?j^?@%f0
Original magnifications: 200 in A, E, G, H; 600 in B, C, D, F. #nMP(ShK
Data are means s.d. of four independent experiments. |Ls&~'ik
*P<0.05 versus 0.5 h. V,"AG
PDGF-BB and -DD, but not PDGF-AA and -CC, induce a downregulation of CCN3 4n.i<K8K[
mRNA as determined by real-time RT-PCR and normalized to %_i0go,^
glyceraldehyde-3-phosphate dehydrogenase mRNA. #h5Hi9LKf
Cell number was counted in duplicate using a Malassez hemocytometer ND99g
Data are means s.d. of four independent experiments. *P<0.05 versus cells treated with ,XF6Xsg2
MCDB media for 24 h, #P<0.05. X\^3,k."
28 XN 0RT>@
The curves are adjusted for history of coronary artery disease, CRP, and progression to N;Gf,pE
ESRD. ~9{-I{=
Both models are independent of age, gender, baseline GFR and proteinuria, and other 9M-]~.O
clinical characteristics and traditional and nontraditional cardiovascular risk factors -%QEzu&
Correlation between serum bone-specific alkaline phosphatase (bAP) and serum PTH (}7o
a9Q<
(n=99, P<0.0001, r2=0.190) j
Wb\"0)
Concentration of serum FGF23 concentration before starting and at the end of the 4-h y^Xxa'y
hemodialysis procedure in 23 patients, expressed as log10 values (n=23, P<0.0001, tX2>a
Student's paired t-test). oR%E_g?mI~
Immunohistochemistry for the hypoxia marker pimonidazole (PIM) and HIF-2 ; I^C
]6D{
arrow=endothelial cell and CD=collecting duct *IfIRR>3l(
Magnification: 1000. eC1c`@C:
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