18 7CvD'QW /
Jurkat cells cultured in calcium-free RPMI-1640 medium (Gibco BRL; number !ZtSbOC '
22300-107) containing calcium-free 10% FBS were triggered by anti-Fas IgM. The ,ul5,ygA
treated cells were harvested at the indicated time points and incubated with Fluo-3-AM at M<8ML!N0;t
a final concentration of 1 micromolar for 30 min at 37°C (Scoltock et al., 2000). The ,8K'F
labeled cells (50,000 cells per treatment) were then analyzed by exciting the cells at 488 A{,n;;
nm and examining the fluorescence emission of Fluo 3 at 530 nm with a FACS Scan, 4\ $
3
(Becton Dickinson). A one micromolar concentration of LPA was used as a positive KI&:9j+M)
control for Ca2+ induction. The data thus obtained was analyzed with the software Win -rY 7)=
MDI 2.8 and represented as contour plots.The effect of chelating intracellular calcium on J?712=9
translocation of annexin I was studied by culturing Jurkat cells in the presence of 10 EZr6oO@Nc
micromolar BAPTA-AM, with or without the addition of anti-Fas IgM. Cells were H:oQ
harvested and fractionated as detailed above, and the S-100 fractions were assessed by #n})X,ip2
immunoblotting for presence or absence of annexin I. c{88m/;eP
Mouse bone marrow transduction and transplantation. SN]LeXesS
Retrovirus-mediated transduction of mouse bone marrow cells was done by published *v #/Y
9}
methods49. Prestimulated low-density bone marrow cells were infected with high-titer Q</h-skLZ
retrovirus supernatant on fibronectin-coated plates. Retrovirus supernatant was generated ku.A|+Tn
in the phoenix-gp cells with a mouse stem cell virus–based retroviral vector coexpressing ()|3
EGFP and HA–RhoH as described50. EGFP+ sorted cells were transplanted by 2cnj@E:5l
intravenous injection into the sublethally irradiated (300 rads with a 137Cs irradiator) Lx+`<<_dJ
Rag2-/- recipient mice. At 9 weeks after transplantation, thymus, peripheral blood, bone IcoowZZ
marrow, spleen and lymph nodes from each recipient mouse were collected for analysis !EFBI+?&
of EGFP+ chimerism and hematopoietic lineage by flow cytometry. Expression of YF[$Q=7.
HA–RhoH and HA–RhoHF73F83 in EGFP+ sorted thymocytes of recipient mice was jXcNAl
confirmed by immunoblot analysis. }: #dV
B+
Determination of renal morphology G(OFr2M
Kidney slices were postfixed in buffered 2% OsO4, dehydrated, and embedded in an ,/o(|sks
Araldite-EM bed 812 mixture. Large sections were cut perpendicular to the renal capsule, {e9Y
!oFg
containing cortex, and medulla. Thin (1 m) sections were analyzed in a blinded manner ^5)_wUf
for morphologic alterations, as previously detailed ;QPy:x3
Patient population YBylyVZ
Patients included in the study met the following criteria: (1) biopsy-proven IMN; (2) zh?4K*>.k
creatinine clearance 30 ml per min per 1.73 m2; and (3) persistent proteinuria >5 g per qX@9N=g`#O
24 h despite treatment with an HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A) VK4/82@5
reductase inhibitor, an ACEi, and/or ARB at maximal tolerated dose for at least 4 months. NV~vuC
The Mayo Institutional Review Board and the Research Ethical Board, University Health iwo$\
Network, University of Toronto approved the study protocol. All patients gave written NhDA7z`b'J
informed consent. Patients who had been on treatment with prednisone, cyclosporine, or )
>$^
wT
19 -A;w$j6*
mycophenolic mofetil within the last 4 months or alkylating agents within the last 6 [Yvsa,2
months were not included in the study. Patients with active infection, diabetes, or a PW^ 8;[\QP
secondary cause of MN (for example, hepatitis B, systemic lupus erythematosus (SLE), m<FK;
medications, malignancies) were also excluded. jZ#UUnR%
Treatment h#O"Q+J9n
At enrollment, a low-sodium (<4 g day-1) and low-protein (0.8 g per kg per day of *H*\gaSh
high-quality protein) diet was recommended and patients were encouraged to maintain g< )72-h
the same diet throughout the duration of the study. All patients received a similar EE9vk*[@C
conservative treatment regimen that included loop diuretics to control edema, an J";=d4Sd
HMG-CoA reductase inhibitor, and an ACEi combined with an ARB if tolerated. Mk "vvk
-Blockers and non-dihydropyridine calcium channel blockers, in that order, were added oT!/J
when required to control systolic blood pressures to <135 mm Hg in >75% of the T0?uC/7H
readings. Patients who after a minimum of 4 months of conservative therapy and kYtHX~@
maximized Ang II blockade had proteinuria >5 g per 24 h received two i.v. infusions of gPp(e
j7
rituximab at a dose of 1000 mg on days 1 and 15. To minimize infusion reactions, )-)pYRlO
patients were premedicated with acetaminophen (1000 mg) and diphenhydramine Q>[Ce3
hydrochloride (50 mg) orally. In addition, methylprednisolone (100 mg, i.v.) was given N*':U^/t4J
prior to the first rituximab infusion. B-cell depletion was defined as CD19+ count >F|qb*Tm7
<5 cells per l at any time and B-cell recovery was defined as CD19+ cell count d}VALjXHX!
>15 cells per l. Patients treated with rituximab, who at month 6 had proteinuria >3 g per w Oj88J)
24 h and in whom CD19+ B-cell counts had increased to >15 cells per l, received a ex=)H%_|
second course of rituximab treatment following the same protocol described above. r
hZQQOQ
Follow-up -O /T?H
In all patients, clinical and laboratory parameters including complete blood counts, ~R$[n.Vpk
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum LhZZc`|7t
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry ysl8LK
and at months 1, 3, 6, 9, and 12. Creatinine clearance and protein and creatinine excretion q>P[n z%
in the urine were assessed by performing two consecutive 24-h urine collections at each !6{Jq]
time point. Data were considered accurate when urinary creatinine excretion was B#Qpd7E+*
consistent with a complete 24 h collection. The mean of the two measurements was p!QR3k.9s
considered for the analysis. The presence of HACAs was evaluated at baseline and at f~U
#z7
months 3, 6, 9, and 12. YK7 \D:
Method / Approach / Study/ Technique i<J^:7
A discussion is presented of a problem-solving system \+G.]|" Y
To improve the efficiency of the method, the following approach may be applied. ,T/GW,?
In order to an investigation was made to find the causes of the 8F\~Wz 7K
Although large collections of rules and equations have been complied, none are generally XRtyC4f
accepted 0c`sb+?
20 -*r
[
This approach will be explained and discussed thoroughly in the body of the report. +<GrRYbC
This can be accomplished by
!,Qm
This algorithm to compute the total cost can be described step by step as follows: ~d&'Lp[3
The above preliminary analysis has provided important information `S5>0r5[
Various methods have been proposed for selecting an optimum... H]VsOr
These concepts have been applied to YfV"_G.ad|
On the basis of the concept mentioned above, ix(U:'{
This can be achieved by KBd7|,j
In addition, tissues were stained for infiltrating lymphocytes (CD20 and CD3), and the ^8b~ZX
amount of interstitial fibrosis was quantified by histomorphometry. Formalin-fixed, QjehDwt|
paraffin-embedded sections were cut onto coated glass slides. Following heat-induced ?OdV1xB
antigen retrieval, sections were incubated at 20°C overnight with either anti-CD20 *iPs4Es-
primary antibody or anti-CD3 primary antibody, both at 1:1000 dilution (Dako, Canada OY+!aG@.
Inc, Mississauga, Ontario, Canada). After rinsing all sections, pretreatment with 3% 2;dM:FHLhO
hydrogen peroxide was performed to prevent endogenous peroxidase activation. Sections [t {vYo
were incubated with a secondary rabbit anti-mouse antibody linked with avidin–biotin FrgW7`s[A
complex. Sections were counterstained with hematoxylin and examined by light A$%@fO.b
microscopy. :yd=No@
The HACA assay is a proprietary bridging enzyme-linked immunosorbent assay l0eANB%Y=@
performed at Genentech Inc. that measures the antibody response to rituximab in human 2qt=jz\s
serum samples.
Tbe_xs^
In all patients, clinical and laboratory parameters including complete blood counts, cjCE3V9X
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum -OHvK0~
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry (3)C_Z
and at months 1, 3, 6, 9, and 12. tpN]evp|
This fact suggests that a new concept nwZ[Ygl|
This was accomplished by taking ... R1]v}f_I"
The preparatory stage is very time consuming process. 0SAG6k~x
Test are performed for validity, completeness, and compatibility h<PYE]?l
There is little hope of achieving successful ... [d!Af4
There has been an increasing awareness of the potential of using most ..so far made have M>`?m
L
not taken this approach, with the exception of uYG #c(lc
Only a few studies can be found. 9OrA
9r
It is a very tedious process to go through eyI-s9#t
It is only when .. has been completed that .. may be effected {jj]K.&
The entire interpretation process is conducted in one's head. =Bcux8wA#6
These approaches are sometimes very tedious. 5^
G7pI7
Several techniques can be used fJSV)\e0
A polynomial parametric model can be written as [the following]/[follows]: avYh\xZ
A xx model is constructed/formulated using xx. {Sd@u$&
A xx model represents an xx by its xx. G
#_(7X&
A process decision model captures the logic essential to e#76h;
From the equation above, xx is equal to the summation of xx times the ... hW*^1%1
21 XnNOj>!
The validity of a xx model can be checked using Euler's formula. Mt7X<?GZm
Given a model, one can mathematically determine whether ... or ... I(iGs I
Equations for xx need to be derived and implemented in the system. &;uGIk>s
A number of heuristic rules have been developed for bi4^ zaCEE
Optimum .. techniques can be made more reliable by ... so that bUipp\[aV
An algorithm based on the characteristic ... is used to determine (_s!,QUe
Euler's formula states the following: Gc3PN
The completed model should agree with the formula. @n~ND).
For manufacturing purposes, a detailed and precise model of the object is necessary ul5::
Engineering design models are very well defined; therefore, A ydy=sj
To keep the domain narrow enough to be implementable, yet wide enough to be useful. K2vPj|
Point of View dxae2 tV
from an implementation standpoint, >E2WZHzd2
From the point of view of this application,
t`&s
From this point of view, Zadeh suggested an inference rule named xxx (CRI for short). [8o
m9
Z3
Information is the meaningful interpretation and correlation of some aggregation of data +wPvQKVfI
in order to allow one to make decisions. k^vmRe<lk
From a practical point of view, the computational aspects of an FLC require a MZhJ,km)
simplification of the fuzzy control algorithm. 9Rt(G_'
The use of a hammer to insert screws, although partly effective, tends to distort, destroy, g'l7Jr3
and generally defeat the purpose of using a screw [Kusiak AI Implications for CIM (t"YoWA#m
p.129] \[hrG?A
Statistical analysis )Lv6vnT>
Data were analyzed by one-way analysis of variance comparing the three conditions [;kj,j
(sham operation, ischemic AKI, and bilateral nephrectomy) at each time point. If =,}!Ns{k
significant F-statistic from analysis of variance existed, this test was followed by Dunnett qkyX*_}
post hoc multiple comparison procedure with sham operation as the control group. For all n$\6}\k
other comparisons, Student's t-test was used. A P-value of <0.05 was considered as xX67bswG
statistically significant. ='l6
&3X
Values are expressed as means s.e.m. and significance was evaluated by Mann–Whitney XQ?)
U test using GraphPad Prism, version 4.0 software (GraphPad Software Inc., San Diego, q=i,'.nS
CA, USA). BM}a?nnoc
All values are expressed as means s.d. Statistical significance (defined as P<0.05) was C8%nBa/
evaluated using analysis of variance and Bonferroni t-tests, and the two-tailed Pearson's jind!@}!
test, where appropriate. '\3.isTsx
Data are expressed as mean s.d., median and interquartile range, or frequencies, as J1wGK|F~
appropriate. Variables who deviated from the normal distribution (positively skewed) )-7(Hv1
were log-transformed (log10) before the correlation study. U S~JLJI
Data are represented as means s.e.m. Student's t-test and multiple comparisons with t-test 9cHNwgD>v
post hoc analysis of variance were used as indicated below, for the comparison of m]b.P,~v
22 (4/`@;[
morphological, immunohistological and functional parameters. Statistical significance #p^pvdvh3
was set at P<0.05. Tr+Y@]"
The primary efficacy parameter was defined as change in urinary protein excretion from l[OQo|_
baseline (week 0) to 12 months after treatment. The 12-month changes were tested =dJEcC_J
against zero using the paired t-test. Secondary end points included 6-month changes in N_~Wu
protein; the number with PR or CR at 6 or 12 months; and changes in glomerular gj(l&F *@
filtration rate (GFR), serum albumin, and lipid profiles. Study sample size was based on ]4pC\0c
the desire to have 80% power to detect a drop in urinary protein of at least 2.0 g day-1. {Eb2<;1o{
Assuming a two-sided hypothesis test performed at a significance level of 0.05 and an s.d. r_2VExk
of urinary protein change of 2.5 g, it was determined that 15 patients were required.2 Op)R3qt{
Definition of remission status is according to the criteria established by Cattran et al.30 GVPEene
CR was defined as proteinuria <0.3 g per 24 h, PR as proteinuria 3 g per 24 h, and a /t2<OU9
>50% reduction in peak proteinuria and non-response as <50% reduction in peak oz7udY=]0
proteinuria. Any patient reaching a CR or PR was considered a treatment success v?iH}7zb%Q
The statistical significance of differences for the mean values of cytokine concentration -)c"cgx.
and T cell proliferation was determined with Student's t-test. Differences with a P value ?J1&,'&
of less than 0.05 were considered significant. 6j.(l4}
Statistical significance was determined by Student's t-test. Data with a P value of less +R jD\6bJb
than 0.05 were considered significant. :1Yd;%>92
In vitro and in vivo experiments were analyzed by the two-tailed Student's t-test, with P - cC(d$y
values less than 0.05 considered statistically significant. rs)aEmvC
The significance of differences among groups was determined by Student's t-test and krT!AfeV
one-way analysis of variance (SigmaStat software; Jandel). ]$%4;o4O
Comparisons -|UX}t*
As well, the pros and cons of these representations from a process planning point of view 8J@OMW&[l
will be discussed. s~'C'B?
The method of using xx to implement xx described by Zadeh (1973) appeared more G\+MT(&5
suitable <{~UKi
As discussed [in the previous section]/[preciously], H_^u_%:e
Relation "PP0PL^5F
We can not invert F' directly because it defines a many-to-one mapping. #8HXR3L5=!
The relationships appear very complicate Or~6t}f
Lifting tasks involve complex and imprecise relationship between the task variables and ]qTr4`.
the human operator's characteristics. `Y$5g~3.
These methods are based on the relationship between ... and ... .EELR]`y7I
The fundamental concept of a fuzzy rating language is that we can establish a relationship >dH*FZ:c
among terms such as high, medium, and low, and then modify these relationships. ho=]'MS|
This article will thus mention the latter as well as the former. +=xRr?F
The former two bear a close relation to a fuzzy Cartesian product. |1 6v4 R
Importance '%7]xp
) jiD8|%}v
23 fXQRsL8
]
The emphasis is on an implementation of a general approach to rule based decision G+p>39P
making. DN_C7\CoA
Consideration / Attention O/~^}8TLL
Careful evaluation is necessary to ensure g0$k_
Such a formulation does not change further considerations. <;"=ah7A
Considerable attention has been paid to tYa*%|!
v
Attention should be paid to an important finding of this investigation. U]$3NIe
Caution should be exercised in this process to avoid ... GjG3aqP&!
Primary consideration is given to ... components, though others can be accommodated #{1fb%L{i
After ... has been defined by ..., a carefully analysis is carried out/performed to determine n-Y'LK40Os
A number of factors such as ...need to be taken into consideration before making the >$F]Ss)$
appropriate decision.
,oil}N(
It should be noted that vL0Ol-Vt
It is important to point out that ... B:.rp.1
These considerations have heightened interest in the possibility of providing ... j+/*NM_y3
We should stress the fundamental importance of the xx l"8YI sir
Results. .|-y+9IP
Advantages / Disadvantage Tkf !Y?
One of the major advantages of this new measure of xx is that it can be applied to the 7m:, -xp
experimental study of \eCdGx?
One advantage of using a .. is the ease of preparing it. !C?z$5g
It has a very fast decision making process b.&YUg[#
All the algorithms involve mostly logical operations. 70eN]OY
It can be easily and without additional cost implemented in a microprocessor;based F^-4Pyq@
environment. Y`uCDfcQ
It can reduce the waste of designing from scratch. *Q?HaG|S
The advantages of using a xx to represent xx are the following: .MlE1n'
However, xx is not without its shortcomings. @T"385>
In most cases, the xxx shows an improvement over the existing xxx. $P_Y8:
Compared to the existing xx, the impacts of the xx are generally reduced by 5% to 9%. \BW(c)Q
The "best case" results shows a savings of 6% to 9%. f`e.c_n(
Most of the existing works based on xx approach can only recognize a xx . 7Z_iQ1
Most of the above methods are computational expansive and limited to xx. &PX'=UT
Some other advantages of xx are the following: &7LfNN`
The problem is the limitation of this method to a limited domain of parts. ,")7uMZaF\
It proved limited in application because it demanded precision in system modeling that ]Ea-?IhD
was impossible in practice. 1Jt%I'C?
There are advantages to be gained in the structuring of costs and benefits, the use of xx, Er)b( Kk
The disadvantages of this method are also disadvantages of conventional xx approaches. 9}N*(PI
This combines the best features of both techniques i{2KMa{K
24 =RA6 p
Hopefully, this tool can be as the reference framework of for developing a xx platform, e]QkZg2?Yn
and helping the administration, marketing, and knowledge management activities in {+;8dtZ)x
virtual communities. W2RS G~|
Results rk|6!kry
An improvement on the result shown above can be made by based on the data provided Z5Cv$bUc
Recent work has identified [>E0(S]
Time-dependent changes of dU,/!|.K
Cyclosporine-induced cell death is triggered by a non-classical phosphoinositide 3-kinase u8*0r{kOH
and does not require ERK activation. LBg#KQ@
Much of the current work on (w*$~p
Discussion of these theories is beyond the scope of this review Rd2[
xk
Based on the information contained in this Vr'Z5F
*@
The result can be categorized into nine classes IH5thL@D
The results are illustrated by an example HwHI$IB
The experimental results for each xx time are reported in Table 2. m5S/T\,X
From the results obtained so far, it seem that \vI_%su1N
Because of the inaccuracy of the ..., a conclusion cannot be drawn as ?@~FT1"6G
Although much effort has been made to., this reality is far from completion. qcK)J/K"
The results indicate that the total benefits are higher than the total costs. =Zj9F1E[i
Their results may then serve as guidelines for lower level models, less fuzzy and more YXvKDw'95
detailed. "783F:mPh
Conclusion <,m}TTq
From the discussion, one may conclude that ... '&"7(8E}
*
The first indication that 6c2fqAF>i
In summary, we have shown that #8PjYB
These results suggested that KT3[{lr
Our findings have shown that ^3el-dZ
Our observations have provided {S|uQgs6j
This study reveals following five main findings: rTi.k
To our knowledge, this is the first immunohistochemical report of both PIM and HIFs in {vaq,2_w
the diabetic kidney. (vQShe\
As mentioned above, 4F'@yi^Gt
Moreover, we demonstrate that, b&LhydaJ
A number of studies have elaborated a body of knowledge about YXmLd'F^3
There are findings to indicate that ~sM334sQ
On a descriptive level, there is agreement that for giI9-
C
One issue that became particularly evident from our study is 3mx7[Q
Our data are in agreement with the findings and models presented by previous cM55
vVd
publications, in particular those of Jones et al l!`m}$
According to our observations, j3$KYf`T}
…was not evident from our data Z
rvb
%
25 tf>?;
It could therefore be concluded that V;>p@uE,P
our data illustrate that %R}}1
All these different aspects, as listed above, lead to 3uN;*f
In fact, it has already been speculated that d~qDQ6!
It should be furthermore mentioned in this context that O%rjY
To identify and verify the essential factors triggering developmental renin expression, it ,;UVQwY
is of interest now to study the development of intrarenal renin expression in mice with la
f b^
defined gene mutations, using the results of this study as a reference system. `^RpT]S
In conclusion, using several approaches, we demonstrated that B1 'Ds
But we have not provided formal proof yet that ,LG6py&aT
In summary, our results suggest that e3&R3{
In conclusion, we identify kAU[lPt*R
A better understanding of such regulatory systems may yield novel therapeutic 7n
{uxE#U)
approaches to glomerular diseases. i{xgygp6f
Preliminary results of short-term studies (2 weeks) indicate that ?PLf+S
However, we believe that LC/6'4}_
However, despite our extensive measurements of these parameters, we found no 0zetOlFbO
relationship between response and initial proteinuria, time to B-cell recovery, the /qEoiL###
development of antibodies to rituximab, B-cell count in the kidney tissue, nor the degree 1%t9ic
of tubular interstitial damage present. A clear conclusion for efficacy, however, cannot be a//<S?d$:
obtained from an uncontrolled study, and definitive claims of efficacy should be reserved pGzzv{H
for rigorous, prospective, controlled, and randomized trials with the use of rituximab and RoT}L#!!
that will also look for factors that may determine its efficacy in IMN. Fep#Pw1
It has become increasingly clear that *ax$R6a#X
Form the above discussion, the conclusion can be reached that "I}3*s9Q-
The conclusions drawn are also valid hxGZ}zq*S
In conclusion to this, it becomes obvious that the problem of xx lies not only in... |-%[Z
We have attempted to introduce some concepts associated with a theory of L
slI!.(
Considerable more work, hopefully, will be done in this area JT#jJ/^
Employing the compositional rule of inference, the assessment of the xx compatibility in p~6/
achieving prescribed xx projectiles in any level of the hierarchy is made possible. 1!~=8FTv
This paper has presented a theoretical and experimental study of the xx process and xx V*"-@
concept. F$F,I,$ "
The experimental research results will hopefully serve as useful feedback information for ~*@UQ9*p#
improvements for xx work. N^jQ\|A<
The scope of this contribution was to introduce a xx method. TFjb1a,)
This has a tremendous impact on our understanding of ;*Ldnj;B
Significant progress has been made in advancing RNAi therapeutics in a remarkably }4b
4<Sm_h
short period of time 5ro^<P0f**
To date, little is known about… ?A+-k4l
Irrespective of the mechanisms and the molecules –which are still largely unknown – #?DwOUw
involved in MSC functions, current data suggest that ^}fc]ovV
To better understand \R\?`8Orz
26 Q'>pOtJG*J
Future Research (常用于conclustion 的结尾)
T)?:q
Thus, first extension of the approach could be, |WwC@3)
Further studies are needed to determine the importance of 0A9llE
Some improvements to the scheduling aspect of the model may be brought through ?5-Y'(r
additional levels in the hierarchy for more detailed representation of the scheduling f9ux+XQk9
activity. G'<:O(Imu
It also unveiled new aspects of the role played by thyroid hormone in response to injury *oZ]k`-!8
and tissue repair. +?
y ', Ir
In the near future, the ongoing clinical trials with siRNAs for macular degeneration and 2t
7':X
RSV may reveal the exciting potential of RNAi therapeutics as the next major class of )B'&XLK
drug molecules. a4aM.o
In the next few years it should become clear whether In13crr4!
It should be an exciting time for researchers seeking to harness this powerful endogenous 2[}
O:
pathway to treat human disease. ;GQCq@)-
Acknowledgement lgrD~Y (x
The authors are grateful to the insightful work provided by Mr. John W. Harney and all 6.5
E
d-
of the fellows and students that contributed with some of the studies discussed in this FJ-X~^
article dmA#v:$1
I would like to express my gratitude to all those who gave me the possibility to complete kA/4W^]Ws
this thesis. |Yb]@9>vn
I want to thank the Department of Transport of the former Interim Government of YOOcHo.F
Namibia for giving me permission to commence this thesis in the first instance, to do the Aq5CF`e{
necessary research work and to use departmental data &6="r}
This work was supported by NIH grants AI058695 and AI056900. We thank members of 2P,%}Ms
the laboratory and our collaborators for many useful suggestions. *RYok{w
We thank J. Maraganore and B. Greene for critical reading of the manuscript, M. i8w/a
Duckman for graphics assistance and A. Capobianco for word processing assistance. ?i{/iH~Sf
Author contributions x3AAn,m8
All authors were involved in experimental planning and data analysis and contributed to B)/L[ )S
manuscript preparation. w`YN#G
Tables and Figures =0t<:-?.-
Figure 7-1 sketches these relationships. 3QI. |;X
The graphical representation of these functions is shown in Figure 1. 1S+lHG92I
The xx may be depicted as in Figure 1. +7\"^D
Figure x shows the schematic diagram of the I|x?
K>
Figure 1 though 2 provide a ... that Y6 <.]H
the architecture of this expert system for .... is illustrated in Figure 2. 6n A/LW\x
Figure 2 gives the outline of an ... system V1#:[o63+
Table shows the DuQW?9^232
27 prEI9/d"
Time-dependent changes of Y{d-k1?s5
as shown in Table 1 and 2 v#|yr<
This concept is illustrated in Figure 2 K+\2cf?bU
At the top of Table xx are shown two blocks of data. 'iWDYZ?
Each table or matrix has constructs xx through xx as row;headings, i@`qam
xx through xx as column;headings. ;w7s>(ITZ
Time course of calculated renin immunoreactive tissue volumes during development of
:al
,zxs
the mouse kidney. Data are single values per time point. They were derived from the )7C+hQe
kidneys shown in Figure 1. o<gK"P
Yellow color indicates )DGJr/)
PCT3 cells were pretreated with 50 M PD98059 or 10 M U0126 for 30 min and then EQtY b"_
cells were stimulated with 25 g ml-1 CsA for 6 h. ERK and PKB activation were a9u2Wlz
analyzed by western blot using phospho-specific antibodies. "m%EFWUOl
ERK and PKB activation were measured by western blot using phospho-specific ~ o=kW2Y
antibodies Zi?:< H}
The data are the means of three different experiments taking the number of untreated cells "
l{{H&d
(vehicle alone) as 100%. q/^?rd
ERK and PKB activation were then analyzed by western blot with the corresponding ||L^yI~_d
phospho-specific antibodies. To ensure equal amount of protein in each lane, western '#C5m#v
blots against total protein were performed for ERK and PKB. ;X;(7
The means of optical density (OD) values of the bands detected in western blots of three F`La_]f?b\
different experiments are plotted, taking the maximal value of phosphorylation of ERK JEs@ky?{z
and PKB as 100%. b<%c ]z
CT3 cells were treated with 25 g ml-1 CsA or vehicle alone for 24 h and then fixed with IhN^*P:Fo
paraformaldehyde and stained with Hoescht as indicated in Materials and Methods. Then S,EL=3},=
cells were visualized by phase-contrast microscopy (upper panel) and fluorescence )HcLpoEi
microscopy (lower panel). cm0$v8
The data are the means of three experiments performed on different days taking the +eO>> ~Z
number of cells at the onset of the experiment as 100%. 7GTDe'T
Rats were killed at 4 h and at days 1, 2, 4, 7, 9, 14, 21, and 28 after disease induction YG /@=Z.
(n=9 each). P4zwTEk`
*P<0.05 versus non-nephritic rats. B<ncOe
The expression of the CCN3 protein was detected by western blot analysis (n=4 each, PC, JVGTmS[3
positive control, M, molecular weight markers). E0x\h<6W~
Original magnifications: 200 in A, E, G, H; 600 in B, C, D, F. }g@
'^v
Data are means s.d. of four independent experiments. :+
mULUi
*P<0.05 versus 0.5 h. ]Ok'C"V(j
PDGF-BB and -DD, but not PDGF-AA and -CC, induce a downregulation of CCN3 EDuH+/:n
mRNA as determined by real-time RT-PCR and normalized to ]28j$)6
glyceraldehyde-3-phosphate dehydrogenase mRNA. r
\[|'hA
Cell number was counted in duplicate using a Malassez hemocytometer 4AKr.a0q
Data are means s.d. of four independent experiments. *P<0.05 versus cells treated with N:CQ$7T{ j
MCDB media for 24 h, #P<0.05. [?hvx}
28 PxQQf I>
The curves are adjusted for history of coronary artery disease, CRP, and progression to ^|=P9'4Th
ESRD. PU[<sr#,
Both models are independent of age, gender, baseline GFR and proteinuria, and other OrYN-A4{
clinical characteristics and traditional and nontraditional cardiovascular risk factors xticC>
Correlation between serum bone-specific alkaline phosphatase (bAP) and serum PTH f,F1k9-1!
(n=99, P<0.0001, r2=0.190) 1C<d^D_!p
Concentration of serum FGF23 concentration before starting and at the end of the 4-h L N
Fe7<y
hemodialysis procedure in 23 patients, expressed as log10 values (n=23, P<0.0001, 3y+~l
H:
Student's paired t-test). S0Io$\ha
Immunohistochemistry for the hypoxia marker pimonidazole (PIM) and HIF-2 ; /a<UKh:A[
arrow=endothelial cell and CD=collecting duct M.6uWwzQR
Magnification: 1000. [^?i<z{0C
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