18 rn$G.SMgz
Jurkat cells cultured in calcium-free RPMI-1640 medium (Gibco BRL; number 6"Ze%:AZZ
22300-107) containing calcium-free 10% FBS were triggered by anti-Fas IgM. The `@q\R-`
treated cells were harvested at the indicated time points and incubated with Fluo-3-AM at iQJa6QF&:
a final concentration of 1 micromolar for 30 min at 37°C (Scoltock et al., 2000). The tk)JE^'
labeled cells (50,000 cells per treatment) were then analyzed by exciting the cells at 488 `q9n`h1
nm and examining the fluorescence emission of Fluo 3 at 530 nm with a FACS Scan, KqN;a i,F
(Becton Dickinson). A one micromolar concentration of LPA was used as a positive 2Z-
[x9t
control for Ca2+ induction. The data thus obtained was analyzed with the software Win s=e`}4
MDI 2.8 and represented as contour plots.The effect of chelating intracellular calcium on jY2mn" .N
translocation of annexin I was studied by culturing Jurkat cells in the presence of 10 D6dliU?k
micromolar BAPTA-AM, with or without the addition of anti-Fas IgM. Cells were (@ 1>G
^%
harvested and fractionated as detailed above, and the S-100 fractions were assessed by fsl
ZJE
immunoblotting for presence or absence of annexin I.
>akC
Mouse bone marrow transduction and transplantation. ?f$U8A4lp
Retrovirus-mediated transduction of mouse bone marrow cells was done by published +7Uv|LZ~@
methods49. Prestimulated low-density bone marrow cells were infected with high-titer 1COSbi]
retrovirus supernatant on fibronectin-coated plates. Retrovirus supernatant was generated n$lVmQ6
in the phoenix-gp cells with a mouse stem cell virus–based retroviral vector coexpressing 4(91T
EGFP and HA–RhoH as described50. EGFP+ sorted cells were transplanted by i[?VF\Y(
intravenous injection into the sublethally irradiated (300 rads with a 137Cs irradiator) ]v/pMg#-
Rag2-/- recipient mice. At 9 weeks after transplantation, thymus, peripheral blood, bone n0LN
AhM
marrow, spleen and lymph nodes from each recipient mouse were collected for analysis EKD#s,(V*X
of EGFP+ chimerism and hematopoietic lineage by flow cytometry. Expression of A^E 6)A=
HA–RhoH and HA–RhoHF73F83 in EGFP+ sorted thymocytes of recipient mice was m68>`
confirmed by immunoblot analysis. Rt5,/Q0
Determination of renal morphology 1:iB1TclP
Kidney slices were postfixed in buffered 2% OsO4, dehydrated, and embedded in an BoQ%QV69%
Araldite-EM bed 812 mixture. Large sections were cut perpendicular to the renal capsule, F [Lg,}
containing cortex, and medulla. Thin (1 m) sections were analyzed in a blinded manner k*u6'IKi.4
for morphologic alterations, as previously detailed cV"Ov@_.k
Patient population 2cO6'?b
Patients included in the study met the following criteria: (1) biopsy-proven IMN; (2) ]61Si~Z
creatinine clearance 30 ml per min per 1.73 m2; and (3) persistent proteinuria >5 g per {+ Ibi{
24 h despite treatment with an HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A) mr2Mu
reductase inhibitor, an ACEi, and/or ARB at maximal tolerated dose for at least 4 months. ?(Ua+*b
The Mayo Institutional Review Board and the Research Ethical Board, University Health J0
dY%pH#
Network, University of Toronto approved the study protocol. All patients gave written )+"5($~
informed consent. Patients who had been on treatment with prednisone, cyclosporine, or ?K;l 5$?%
19 f ~*7hv\
mycophenolic mofetil within the last 4 months or alkylating agents within the last 6 -uu&{$
months were not included in the study. Patients with active infection, diabetes, or a W"*2,R[}%
secondary cause of MN (for example, hepatitis B, systemic lupus erythematosus (SLE), QF
npp\K
medications, malignancies) were also excluded. Q)y5'u qZ
Treatment "G-h8IN^O
At enrollment, a low-sodium (<4 g day-1) and low-protein (0.8 g per kg per day of u;]xAr1
high-quality protein) diet was recommended and patients were encouraged to maintain k$ T
the same diet throughout the duration of the study. All patients received a similar Gov.;
hy
conservative treatment regimen that included loop diuretics to control edema, an 7p hf
HMG-CoA reductase inhibitor, and an ACEi combined with an ARB if tolerated. +zd/<
-Blockers and non-dihydropyridine calcium channel blockers, in that order, were added 8MI8~
when required to control systolic blood pressures to <135 mm Hg in >75% of the ,[T/O\k
readings. Patients who after a minimum of 4 months of conservative therapy and dA (n,@{
maximized Ang II blockade had proteinuria >5 g per 24 h received two i.v. infusions of 3z, Ci$[
rituximab at a dose of 1000 mg on days 1 and 15. To minimize infusion reactions, ZjMnGRP
patients were premedicated with acetaminophen (1000 mg) and diphenhydramine a|=^
hydrochloride (50 mg) orally. In addition, methylprednisolone (100 mg, i.v.) was given ^-Ygh[x
prior to the first rituximab infusion. B-cell depletion was defined as CD19+ count S 6_:\Q
<5 cells per l at any time and B-cell recovery was defined as CD19+ cell count <YtjE!2
>15 cells per l. Patients treated with rituximab, who at month 6 had proteinuria >3 g per 122s7A
24 h and in whom CD19+ B-cell counts had increased to >15 cells per l, received a oPF]]Imu
second course of rituximab treatment following the same protocol described above. $|@vmv0
Follow-up \v6lcAL-
In all patients, clinical and laboratory parameters including complete blood counts, {l5fKVb\C
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum },#7
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry @{q<"hT
and at months 1, 3, 6, 9, and 12. Creatinine clearance and protein and creatinine excretion [>r0
(x&.
in the urine were assessed by performing two consecutive 24-h urine collections at each {:("oK6w
time point. Data were considered accurate when urinary creatinine excretion was $H[q5(_~
consistent with a complete 24 h collection. The mean of the two measurements was oSmETk\
considered for the analysis. The presence of HACAs was evaluated at baseline and at ,OubKcNg
months 3, 6, 9, and 12. 9(q(;|;Hp
Method / Approach / Study/ Technique 9]w?mHslE
A discussion is presented of a problem-solving system c AO:
fb7
To improve the efficiency of the method, the following approach may be applied. ls&H oJ7
In order to an investigation was made to find the causes of the mB"I(>q*M
Although large collections of rules and equations have been complied, none are generally r~h#
accepted +)WU:aKI
20 {4,],0bjx/
This approach will be explained and discussed thoroughly in the body of the report. H!Q72tyo
This can be accomplished by ;>
jEeIlT
This algorithm to compute the total cost can be described step by step as follows: qD ?`Yd
The above preliminary analysis has provided important information Gj-nTN
Various methods have been proposed for selecting an optimum... ;*<R~HJt
These concepts have been applied to 'UWkJ2:!
On the basis of the concept mentioned above, (@5`beEd
This can be achieved by OVDuF&0
In addition, tissues were stained for infiltrating lymphocytes (CD20 and CD3), and the 6(d6Uwc`
amount of interstitial fibrosis was quantified by histomorphometry. Formalin-fixed, 6V]m0{:E
paraffin-embedded sections were cut onto coated glass slides. Following heat-induced 4jC)"tch
antigen retrieval, sections were incubated at 20°C overnight with either anti-CD20 &`v?oN9$
primary antibody or anti-CD3 primary antibody, both at 1:1000 dilution (Dako, Canada kl.; E{PL
Inc, Mississauga, Ontario, Canada). After rinsing all sections, pretreatment with 3% 'WE"$1
hydrogen peroxide was performed to prevent endogenous peroxidase activation. Sections 3MNM<Ih
were incubated with a secondary rabbit anti-mouse antibody linked with avidin–biotin C'|9nK$%
complex. Sections were counterstained with hematoxylin and examined by light -'d:~:1f
microscopy. s.
A}ydtt
The HACA assay is a proprietary bridging enzyme-linked immunosorbent assay e~zgH\`
performed at Genentech Inc. that measures the antibody response to rituximab in human 4BCe;Q^6
serum samples. ga%\n!S
In all patients, clinical and laboratory parameters including complete blood counts, w=WF$)ZU
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum s __xBY
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry O%\cRn8m
and at months 1, 3, 6, 9, and 12. ftxL-7y
%
This fact suggests that a new concept MVzuE}
This was accomplished by taking ... GT<oYrjU
The preparatory stage is very time consuming process. 8A3/@Z;0S
Test are performed for validity, completeness, and compatibility F5
]<=i
There is little hope of achieving successful ... } jJKE
There has been an increasing awareness of the potential of using most ..so far made have
Q*>)W{H&)
not taken this approach, with the exception of *7h~0%WR
Only a few studies can be found. OA3J(4!"W
It is a very tedious process to go through b`#YJpA
It is only when .. has been completed that .. may be effected js81@WX!c
The entire interpretation process is conducted in one's head. hXjZ>n``
These approaches are sometimes very tedious. BeVDTk:
Several techniques can be used kPs?
A polynomial parametric model can be written as [the following]/[follows]: uH9Vj<E$K
A xx model is constructed/formulated using xx.
= ){G
A xx model represents an xx by its xx. /fp8tL2Y
A process decision model captures the logic essential to XMI*obS'z
From the equation above, xx is equal to the summation of xx times the ... k=mQG~
21 E#,"C`&*
The validity of a xx model can be checked using Euler's formula. cRnDAn#42
Given a model, one can mathematically determine whether ... or ... dRron_'
Equations for xx need to be derived and implemented in the system. :.*Q@X}-I
A number of heuristic rules have been developed for c6xr[tc%
Optimum .. techniques can be made more reliable by ... so that Og7yT{h_
An algorithm based on the characteristic ... is used to determine /(BS<A
Euler's formula states the following: SF:98#pg
The completed model should agree with the formula. VAL?
Z
For manufacturing purposes, a detailed and precise model of the object is necessary 6LDZ|K@
Engineering design models are very well defined; therefore, hH1Q:}a
To keep the domain narrow enough to be implementable, yet wide enough to be useful. h.vy SwF"j
Point of View utZI'5i
from an implementation standpoint, V|'@D#\
From the point of view of this application, \|Af26
From this point of view, Zadeh suggested an inference rule named xxx (CRI for short). ElW\;C:K*
Information is the meaningful interpretation and correlation of some aggregation of data e` 4mrBtz|
in order to allow one to make decisions. +yO) 3
From a practical point of view, the computational aspects of an FLC require a #'&-S@/nQs
simplification of the fuzzy control algorithm. w
PR Ns9^
The use of a hammer to insert screws, although partly effective, tends to distort, destroy, QPf\lN/$4d
and generally defeat the purpose of using a screw [Kusiak AI Implications for CIM W8 g13oAu"
p.129] 1!1JT;gG^9
Statistical analysis :5~Dca_iU4
Data were analyzed by one-way analysis of variance comparing the three conditions ,h^6y
(sham operation, ischemic AKI, and bilateral nephrectomy) at each time point. If nN{DO:_o
significant F-statistic from analysis of variance existed, this test was followed by Dunnett bDvGFSAH
post hoc multiple comparison procedure with sham operation as the control group. For all d
8:
$ll
other comparisons, Student's t-test was used. A P-value of <0.05 was considered as nr]=O`Mvh
statistically significant. En&7 e
Values are expressed as means s.e.m. and significance was evaluated by Mann–Whitney q<E7qY+
U test using GraphPad Prism, version 4.0 software (GraphPad Software Inc., San Diego, wetu.aMp
CA, USA). D/=05E%[81
All values are expressed as means s.d. Statistical significance (defined as P<0.05) was [1(eSH
evaluated using analysis of variance and Bonferroni t-tests, and the two-tailed Pearson's cY!Y?O
test, where appropriate. aZ~e;}w.Zq
Data are expressed as mean s.d., median and interquartile range, or frequencies, as N#M>2b<A/T
appropriate. Variables who deviated from the normal distribution (positively skewed) Mt4]\pMUb
were log-transformed (log10) before the correlation study. =
~O3j:<6
Data are represented as means s.e.m. Student's t-test and multiple comparisons with t-test 2Di~}* 9&
post hoc analysis of variance were used as indicated below, for the comparison of |5;,]lbt
22 w-WAgAch
morphological, immunohistological and functional parameters. Statistical significance ! =21K0~t#
was set at P<0.05. ek)rsxf1A
The primary efficacy parameter was defined as change in urinary protein excretion from <IR@/b!,
baseline (week 0) to 12 months after treatment. The 12-month changes were tested %.
((4 6)
against zero using the paired t-test. Secondary end points included 6-month changes in A(&\wd
protein; the number with PR or CR at 6 or 12 months; and changes in glomerular \&vXp"-@
filtration rate (GFR), serum albumin, and lipid profiles. Study sample size was based on ]Yt3@ug_f
the desire to have 80% power to detect a drop in urinary protein of at least 2.0 g day-1. |6-9vU!LK?
Assuming a two-sided hypothesis test performed at a significance level of 0.05 and an s.d. K_U`T;Z\
of urinary protein change of 2.5 g, it was determined that 15 patients were required.2 }%wP^6G*x\
Definition of remission status is according to the criteria established by Cattran et al.30 ,eebO~7vB
CR was defined as proteinuria <0.3 g per 24 h, PR as proteinuria 3 g per 24 h, and a ;,Vdj[W$>
>50% reduction in peak proteinuria and non-response as <50% reduction in peak * g+v*q X
proteinuria. Any patient reaching a CR or PR was considered a treatment success <*iFVjSI(
The statistical significance of differences for the mean values of cytokine concentration mu0L_u(P
and T cell proliferation was determined with Student's t-test. Differences with a P value q#Otp\f
of less than 0.05 were considered significant. !pe[H*Cy
Statistical significance was determined by Student's t-test. Data with a P value of less <2j$P Y9
than 0.05 were considered significant. 2FL_!;p;2E
In vitro and in vivo experiments were analyzed by the two-tailed Student's t-test, with P 5D3&E_S
values less than 0.05 considered statistically significant. )+G0m,n
The significance of differences among groups was determined by Student's t-test and Pi[]k]XA\
one-way analysis of variance (SigmaStat software; Jandel). n"iaE
Comparisons SN[yC
As well, the pros and cons of these representations from a process planning point of view i{:?Iw 'ay
will be discussed. ,-d2wzhW
The method of using xx to implement xx described by Zadeh (1973) appeared more k!qOE\%B
suitable
aG"
As discussed [in the previous section]/[preciously], Lo'GfHE
Relation x6T$HN/2
We can not invert F' directly because it defines a many-to-one mapping. LfnQcI$kO
The relationships appear very complicate T}p|_)&y
Lifting tasks involve complex and imprecise relationship between the task variables and 6EWCJ%_
the human operator's characteristics. T?n [1%K
These methods are based on the relationship between ... and ... C>l (4*S
The fundamental concept of a fuzzy rating language is that we can establish a relationship M(^IRI-
among terms such as high, medium, and low, and then modify these relationships. I`{=[.c
This article will thus mention the latter as well as the former. 99zMdo S
The former two bear a close relation to a fuzzy Cartesian product. O> wGJ.
Importance ) \
cnz
) N:^4OnVR
23
A[88IMZs
The emphasis is on an implementation of a general approach to rule based decision usugjx^p
making. sGMnm
Consideration / Attention /{ 8 .Jcx$
Careful evaluation is necessary to ensure Df$Yn
Such a formulation does not change further considerations. wzI*QXV2s
Considerable attention has been paid to [/cJc%{N
Attention should be paid to an important finding of this investigation. -fz( ]d
Caution should be exercised in this process to avoid ... <dY{@Cgw=
Primary consideration is given to ... components, though others can be accommodated >&qaT*_g
After ... has been defined by ..., a carefully analysis is carried out/performed to determine No/D"S#
A number of factors such as ...need to be taken into consideration before making the JZNvuP D
appropriate decision. G>^ _&(c@2
It should be noted that w4&-9[@Y
It is important to point out that ... f2$<4Hhmm
These considerations have heightened interest in the possibility of providing ... SQ`ec95',
We should stress the fundamental importance of the xx +m6acu)N.
Results. B{_-k
Advantages / Disadvantage h3-dJgb
One of the major advantages of this new measure of xx is that it can be applied to the iHOvCrp+X
experimental study of 4vPKDd
One advantage of using a .. is the ease of preparing it. 7!,
p,|K
It has a very fast decision making process }Mh`j$
All the algorithms involve mostly logical operations. I#t#%!InH
It can be easily and without additional cost implemented in a microprocessor;based _k5-Wd5Ypw
environment. q}7(w$&
It can reduce the waste of designing from scratch. w259':
The advantages of using a xx to represent xx are the following: E@ !~q
However, xx is not without its shortcomings. m[*y9A1
In most cases, the xxx shows an improvement over the existing xxx. fxX4
!r
Compared to the existing xx, the impacts of the xx are generally reduced by 5% to 9%. /T/7O
The "best case" results shows a savings of 6% to 9%. @$d_JwI
Most of the existing works based on xx approach can only recognize a xx . qgLj^{
Most of the above methods are computational expansive and limited to xx. !xZ`()D#
Some other advantages of xx are the following: $P~Tt 4068
The problem is the limitation of this method to a limited domain of parts. 5-X(K 'Q
It proved limited in application because it demanded precision in system modeling that aruT eJF
was impossible in practice. C")NNs=
There are advantages to be gained in the structuring of costs and benefits, the use of xx, %Nm69j-5%
The disadvantages of this method are also disadvantages of conventional xx approaches. G-Ju`.
This combines the best features of both techniques c1Ta!p{%
24 n*fsdo~
Hopefully, this tool can be as the reference framework of for developing a xx platform, N,NEg4 q[
and helping the administration, marketing, and knowledge management activities in 0E#3XhU
virtual communities. \?.Tq24
Results H
GRH9W
An improvement on the result shown above can be made by based on the data provided b:}+l;e52
Recent work has identified q+-Bl
Time-dependent changes of O(QJiS
Cyclosporine-induced cell death is triggered by a non-classical phosphoinositide 3-kinase gkyv[
and does not require ERK activation. Fps.Fhm
Much of the current work on u?n{r
Discussion of these theories is beyond the scope of this review & ]/Z~V t
Based on the information contained in this 9fL48f$
The result can be categorized into nine classes B}y-zj;T
The results are illustrated by an example ^g56:j~?
The experimental results for each xx time are reported in Table 2. #I.Wmfz
From the results obtained so far, it seem that Eo }mSd
Because of the inaccuracy of the ..., a conclusion cannot be drawn as x
p#+{}
Although much effort has been made to., this reality is far from completion. |F 18j9
The results indicate that the total benefits are higher than the total costs. Qm\VZ<6/5
Their results may then serve as guidelines for lower level models, less fuzzy and more <2a7>\74E0
detailed. CdRgI^5
Conclusion `)W}4itm
From the discussion, one may conclude that ... Rg6e7JVu
The first indication that M/,jHG8v
In summary, we have shown that
PiRbdl
These results suggested that G*|2qX"o
Our findings have shown that i}5
#n
Our observations have provided b)A$lP%`
This study reveals following five main findings: v?7.)2XcX
To our knowledge, this is the first immunohistochemical report of both PIM and HIFs in h.6yI
the diabetic kidney. F:CqB
|
As mentioned above, %@{);5[
Moreover, we demonstrate that, ~pqp`
A number of studies have elaborated a body of knowledge about <o_H]c->
There are findings to indicate that Cp_YIcnEJ
On a descriptive level, there is agreement that for >4.{|0%ut
One issue that became particularly evident from our study is #GYCU!
Our data are in agreement with the findings and models presented by previous w
K[xLf
publications, in particular those of Jones et al 3H%o
TgWk
According to our observations, (s,*soAN
…was not evident from our data rBP!RSl1
25 mB$r>G/'
It could therefore be concluded that TZq']Z)#
our data illustrate that 6M"]p
All these different aspects, as listed above, lead to $H/3t? 6h`
In fact, it has already been speculated that *N$#cz
It should be furthermore mentioned in this context that E#_TX3B
To identify and verify the essential factors triggering developmental renin expression, it 'hL\xf{
is of interest now to study the development of intrarenal renin expression in mice with !9i,V{$c`"
defined gene mutations, using the results of this study as a reference system. +EcN[-~
In conclusion, using several approaches, we demonstrated that Q^8C*ekfg!
But we have not provided formal proof yet that nVoPTr
In summary, our results suggest that :JSOj@s
In conclusion, we identify p~28?lYv
A better understanding of such regulatory systems may yield novel therapeutic vcAs!ls+
approaches to glomerular diseases. R\+p`n$
Preliminary results of short-term studies (2 weeks) indicate that e.i5j^5u
However, we believe that TE0hVw0c
However, despite our extensive measurements of these parameters, we found no .8CR
\-
relationship between response and initial proteinuria, time to B-cell recovery, the oa+Rr&t'
development of antibodies to rituximab, B-cell count in the kidney tissue, nor the degree 7
#_{UJ%
of tubular interstitial damage present. A clear conclusion for efficacy, however, cannot be ]]+wDhxH
obtained from an uncontrolled study, and definitive claims of efficacy should be reserved {y'c*NS
for rigorous, prospective, controlled, and randomized trials with the use of rituximab and w/N.#s^
that will also look for factors that may determine its efficacy in IMN. q?&vV`PG5
It has become increasingly clear that C]-Z+9Vvv
Form the above discussion, the conclusion can be reached that Rw*l#cr=.
The conclusions drawn are also valid H__9%p#
In conclusion to this, it becomes obvious that the problem of xx lies not only in... y-{^L`%Mk
We have attempted to introduce some concepts associated with a theory of +is;$1rq
Considerable more work, hopefully, will be done in this area ./)A6O*#
Employing the compositional rule of inference, the assessment of the xx compatibility in 9rtcI[&?0
achieving prescribed xx projectiles in any level of the hierarchy is made possible. 6Ez}
A|i
This paper has presented a theoretical and experimental study of the xx process and xx %K
/=7
concept. ;58l_ue
The experimental research results will hopefully serve as useful feedback information for 4)-LlYS_d<
improvements for xx work. _{KQQ
5k\
The scope of this contribution was to introduce a xx method. >tqLwC."'
This has a tremendous impact on our understanding of 1/3Go97/qV
Significant progress has been made in advancing RNAi therapeutics in a remarkably '
V;cA$ $
short period of time @X"p"3V
To date, little is known about… g` 6Xrf
Irrespective of the mechanisms and the molecules –which are still largely unknown – NHUx-IqOX
involved in MSC functions, current data suggest that \q(RqD
To better understand 4_8%ZaQ\.?
26 t*<.^+Vd
Future Research (常用于conclustion 的结尾) Xp{+){Iu
Thus, first extension of the approach could be, S>aN#
Further studies are needed to determine the importance of Z'>Xn^
Some improvements to the scheduling aspect of the model may be brought through jtV{Lf3<
additional levels in the hierarchy for more detailed representation of the scheduling .:*V
CDOM
activity. lKrD.iYt8
It also unveiled new aspects of the role played by thyroid hormone in response to injury .KGW#Qk8
and tissue repair. O$E3ry+?
In the near future, the ongoing clinical trials with siRNAs for macular degeneration and >0=` 3X|Y7
RSV may reveal the exciting potential of RNAi therapeutics as the next major class of <bWhTNOb
drug molecules. vAbMU
In the next few years it should become clear whether l!<Nw8+U
It should be an exciting time for researchers seeking to harness this powerful endogenous 3205gI,
pathway to treat human disease. @
|6n.'f+
Acknowledgement ><viJ$i
The authors are grateful to the insightful work provided by Mr. John W. Harney and all @mW: FVI
of the fellows and students that contributed with some of the studies discussed in this KcglpKV`
article {j+w|;
dZF
I would like to express my gratitude to all those who gave me the possibility to complete O7I:Y85i#O
this thesis. E9;cd$}K
I want to thank the Department of Transport of the former Interim Government of 6n]fr9f
Namibia for giving me permission to commence this thesis in the first instance, to do the .dt7b4.kd
necessary research work and to use departmental data &/d;4Eu
This work was supported by NIH grants AI058695 and AI056900. We thank members of "GEJ9_a[
the laboratory and our collaborators for many useful suggestions. N0oBtGb
We thank J. Maraganore and B. Greene for critical reading of the manuscript, M. ='q:Io?T
Duckman for graphics assistance and A. Capobianco for word processing assistance. <LIL{g0eX
Author contributions )d!,,o
All authors were involved in experimental planning and data analysis and contributed to Zj99]4?9
manuscript preparation. \Y_2Z/
Tables and Figures ?~$0;5)QC
Figure 7-1 sketches these relationships. "~0m_brf
The graphical representation of these functions is shown in Figure 1. -Mx"ox
The xx may be depicted as in Figure 1. ?D]4*qsIlu
Figure x shows the schematic diagram of the rn #FmM
Figure 1 though 2 provide a ... that Q3vC^}Dmr
the architecture of this expert system for .... is illustrated in Figure 2. 0tv"tA;
Figure 2 gives the outline of an ... system m_\w)
Table shows the N@lTn}U
27 zs<W>gBq
Time-dependent changes of NBuibL
as shown in Table 1 and 2 zV_U/]y
This concept is illustrated in Figure 2 [,Q(~Qb
At the top of Table xx are shown two blocks of data. (xf_
Each table or matrix has constructs xx through xx as row;headings, 0)`{]&
xx through xx as column;headings. HgPRz C
Time course of calculated renin immunoreactive tissue volumes during development of REUWK#>
the mouse kidney. Data are single values per time point. They were derived from the .[%em9u
kidneys shown in Figure 1. 9ot A5I^v
Yellow color indicates j&6'sg;n)
PCT3 cells were pretreated with 50 M PD98059 or 10 M U0126 for 30 min and then ++d(}^C;
cells were stimulated with 25 g ml-1 CsA for 6 h. ERK and PKB activation were 0g}+%5]yg
analyzed by western blot using phospho-specific antibodies. 20750G
ERK and PKB activation were measured by western blot using phospho-specific F(c~D0
antibodies gKy@$at&
The data are the means of three different experiments taking the number of untreated cells tgX
Ij5z
(vehicle alone) as 100%. c6FKpdn%
ERK and PKB activation were then analyzed by western blot with the corresponding Rp$t;=SMD
phospho-specific antibodies. To ensure equal amount of protein in each lane, western m99j]wr~c
blots against total protein were performed for ERK and PKB. l*_%K}%?V
The means of optical density (OD) values of the bands detected in western blots of three t)P5bQ+$u9
different experiments are plotted, taking the maximal value of phosphorylation of ERK bh9rsRb}O
and PKB as 100%. XDs )
CT3 cells were treated with 25 g ml-1 CsA or vehicle alone for 24 h and then fixed with 2
<HG=iSf
paraformaldehyde and stained with Hoescht as indicated in Materials and Methods. Then n!=%MgF'*p
cells were visualized by phase-contrast microscopy (upper panel) and fluorescence e FDhJ
microscopy (lower panel). jY>|>]4X
The data are the means of three experiments performed on different days taking the V?AHj<
number of cells at the onset of the experiment as 100%. !R*%F
Rats were killed at 4 h and at days 1, 2, 4, 7, 9, 14, 21, and 28 after disease induction *fyC@fI>
(n=9 each). 3]xe7F'`
*P<0.05 versus non-nephritic rats. 'PPVM@)fU
The expression of the CCN3 protein was detected by western blot analysis (n=4 each, PC, %\6ns
positive control, M, molecular weight markers). ~XAtt\WS
Original magnifications: 200 in A, E, G, H; 600 in B, C, D, F. 0sq?;~U
Data are means s.d. of four independent experiments. XQL"D)fw
*P<0.05 versus 0.5 h. KWtLrZ(j
PDGF-BB and -DD, but not PDGF-AA and -CC, induce a downregulation of CCN3 &"GHD{ix
mRNA as determined by real-time RT-PCR and normalized to *>Sb4:
glyceraldehyde-3-phosphate dehydrogenase mRNA. '5xf?0@s.
Cell number was counted in duplicate using a Malassez hemocytometer xbJ@ z{
Data are means s.d. of four independent experiments. *P<0.05 versus cells treated with 7tWt3
MCDB media for 24 h, #P<0.05. 9F+i+(\,b
28
E~oQ%X~
The curves are adjusted for history of coronary artery disease, CRP, and progression to |jB]5ciT
ESRD. `L<
f15][
Both models are independent of age, gender, baseline GFR and proteinuria, and other VsS.\1
clinical characteristics and traditional and nontraditional cardiovascular risk factors |lH~nU.*
Correlation between serum bone-specific alkaline phosphatase (bAP) and serum PTH r~q*E'n
(n=99, P<0.0001, r2=0.190) Mazjn?f
Concentration of serum FGF23 concentration before starting and at the end of the 4-h RqTW$94RD
hemodialysis procedure in 23 patients, expressed as log10 values (n=23, P<0.0001, ""`>v`\
Student's paired t-test).
p x;X}Cd
Immunohistochemistry for the hypoxia marker pimonidazole (PIM) and HIF-2 ; L \0nO i
arrow=endothelial cell and CD=collecting duct Jej P91
Magnification: 1000. fsc^8
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