18 9 wSl,B-
Jurkat cells cultured in calcium-free RPMI-1640 medium (Gibco BRL; number
;# R3k
22300-107) containing calcium-free 10% FBS were triggered by anti-Fas IgM. The nszpG1U:
treated cells were harvested at the indicated time points and incubated with Fluo-3-AM at i>tW|N
a final concentration of 1 micromolar for 30 min at 37°C (Scoltock et al., 2000). The } g3HoFC
labeled cells (50,000 cells per treatment) were then analyzed by exciting the cells at 488 FX|0R#4vm
nm and examining the fluorescence emission of Fluo 3 at 530 nm with a FACS Scan, ub`z7gL
(Becton Dickinson). A one micromolar concentration of LPA was used as a positive gro7*<
control for Ca2+ induction. The data thus obtained was analyzed with the software Win ->&BcPLn
MDI 2.8 and represented as contour plots.The effect of chelating intracellular calcium on DL'd&;6
translocation of annexin I was studied by culturing Jurkat cells in the presence of 10 @9_nwf~X4
micromolar BAPTA-AM, with or without the addition of anti-Fas IgM. Cells were ur/Oc24i1n
harvested and fractionated as detailed above, and the S-100 fractions were assessed by O%YjWb
immunoblotting for presence or absence of annexin I. S}zC3
Mouse bone marrow transduction and transplantation. 5wzQ?07T_
Retrovirus-mediated transduction of mouse bone marrow cells was done by published =gW"#ZjL){
methods49. Prestimulated low-density bone marrow cells were infected with high-titer Qy{NS.T
retrovirus supernatant on fibronectin-coated plates. Retrovirus supernatant was generated &q4ox7 1
in the phoenix-gp cells with a mouse stem cell virus–based retroviral vector coexpressing JZ %`%rA
EGFP and HA–RhoH as described50. EGFP+ sorted cells were transplanted by &e%{k@
intravenous injection into the sublethally irradiated (300 rads with a 137Cs irradiator) Xy[}G p
Rag2-/- recipient mice. At 9 weeks after transplantation, thymus, peripheral blood, bone !(B_EM
marrow, spleen and lymph nodes from each recipient mouse were collected for analysis xM% H~(
of EGFP+ chimerism and hematopoietic lineage by flow cytometry. Expression of y!S^xS
HA–RhoH and HA–RhoHF73F83 in EGFP+ sorted thymocytes of recipient mice was +7Sf8tg\
confirmed by immunoblot analysis. `ifb<T
Determination of renal morphology I_:t}3s
Kidney slices were postfixed in buffered 2% OsO4, dehydrated, and embedded in an _mw13jcN]
Araldite-EM bed 812 mixture. Large sections were cut perpendicular to the renal capsule, Lqg]Fd
containing cortex, and medulla. Thin (1 m) sections were analyzed in a blinded manner gB>(xY>LrA
for morphologic alterations, as previously detailed 1lbwJVY[
Patient population $RpFxi
Patients included in the study met the following criteria: (1) biopsy-proven IMN; (2) et@">D%;]
creatinine clearance 30 ml per min per 1.73 m2; and (3) persistent proteinuria >5 g per ,(6)ghr
24 h despite treatment with an HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A) +IvNyj|
reductase inhibitor, an ACEi, and/or ARB at maximal tolerated dose for at least 4 months. jh>N_cp
The Mayo Institutional Review Board and the Research Ethical Board, University Health eUYZxe :6
Network, University of Toronto approved the study protocol. All patients gave written sD{j@WEZ
informed consent. Patients who had been on treatment with prednisone, cyclosporine, or yXkt:O,i
19 { AFf:[G
mycophenolic mofetil within the last 4 months or alkylating agents within the last 6 d60c$?"]a(
months were not included in the study. Patients with active infection, diabetes, or a 1y7y0V
secondary cause of MN (for example, hepatitis B, systemic lupus erythematosus (SLE), )5U!>,fT
medications, malignancies) were also excluded. &i)helXs]
Treatment X2LV&oi
At enrollment, a low-sodium (<4 g day-1) and low-protein (0.8 g per kg per day of uN8/Q2
high-quality protein) diet was recommended and patients were encouraged to maintain DjY8nePyE
the same diet throughout the duration of the study. All patients received a similar Sg_O?.
r
conservative treatment regimen that included loop diuretics to control edema, an iM
"asEU
HMG-CoA reductase inhibitor, and an ACEi combined with an ARB if tolerated. +N7"EROc
-Blockers and non-dihydropyridine calcium channel blockers, in that order, were added \Y+")
when required to control systolic blood pressures to <135 mm Hg in >75% of the o OQ'*7_
readings. Patients who after a minimum of 4 months of conservative therapy and fa4=h;>a+
maximized Ang II blockade had proteinuria >5 g per 24 h received two i.v. infusions of &f"T,4Oh
rituximab at a dose of 1000 mg on days 1 and 15. To minimize infusion reactions, b;*c:{W)
patients were premedicated with acetaminophen (1000 mg) and diphenhydramine Xo\S9,s{
hydrochloride (50 mg) orally. In addition, methylprednisolone (100 mg, i.v.) was given 9)Ly}Kzx
prior to the first rituximab infusion. B-cell depletion was defined as CD19+ count 2[ksi51y
<5 cells per l at any time and B-cell recovery was defined as CD19+ cell count =HS4I.@c_5
>15 cells per l. Patients treated with rituximab, who at month 6 had proteinuria >3 g per jQ|:I7y
24 h and in whom CD19+ B-cell counts had increased to >15 cells per l, received a eIJ[0c b}
second course of rituximab treatment following the same protocol described above. IT18v[-G
Follow-up (-(QDRxK
In all patients, clinical and laboratory parameters including complete blood counts, o<rsAe
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum `BjR.xMv
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry Pj&A=
and at months 1, 3, 6, 9, and 12. Creatinine clearance and protein and creatinine excretion {[P!$
/
in the urine were assessed by performing two consecutive 24-h urine collections at each 1O0X-C,wo$
time point. Data were considered accurate when urinary creatinine excretion was @vt$MiOi
consistent with a complete 24 h collection. The mean of the two measurements was l `R KqT+
considered for the analysis. The presence of HACAs was evaluated at baseline and at G?ig1PB"#
months 3, 6, 9, and 12. 'yr{^Pek
Method / Approach / Study/ Technique $@.jZ_G
A discussion is presented of a problem-solving system NbkK&bz
To improve the efficiency of the method, the following approach may be applied. jT-<IJh!o
In order to an investigation was made to find the causes of the `{o$F ::(
Although large collections of rules and equations have been complied, none are generally p9iu:MucD<
accepted ~},W8\C>
20 AY88h$a
This approach will be explained and discussed thoroughly in the body of the report. CA*~2|
This can be accomplished by g "*;nHI D
This algorithm to compute the total cost can be described step by step as follows: mNDuwDd$S
The above preliminary analysis has provided important information ="(>>C1-
Various methods have been proposed for selecting an optimum... .iT4-
These concepts have been applied to g~)3WfC$[
On the basis of the concept mentioned above, ^l(^z fsZ
This can be achieved by B^q<2S;
In addition, tissues were stained for infiltrating lymphocytes (CD20 and CD3), and the <5IQc[3]aP
amount of interstitial fibrosis was quantified by histomorphometry. Formalin-fixed, [h-norB((
paraffin-embedded sections were cut onto coated glass slides. Following heat-induced Pm?B
9S
antigen retrieval, sections were incubated at 20°C overnight with either anti-CD20 wS$46M<
primary antibody or anti-CD3 primary antibody, both at 1:1000 dilution (Dako, Canada ]w[ThHRJ
Inc, Mississauga, Ontario, Canada). After rinsing all sections, pretreatment with 3% N
!c
gN
hydrogen peroxide was performed to prevent endogenous peroxidase activation. Sections hY=I5[*
were incubated with a secondary rabbit anti-mouse antibody linked with avidin–biotin vr4S9`,
complex. Sections were counterstained with hematoxylin and examined by light !^v\^Fc
microscopy. +)gB9DoK
The HACA assay is a proprietary bridging enzyme-linked immunosorbent assay m
=k%,J_
performed at Genentech Inc. that measures the antibody response to rituximab in human 2C1+_IL
serum samples. o;QZe&
In all patients, clinical and laboratory parameters including complete blood counts, ~ 5"J(
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum w+_Wc~f
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry ym%o}(v-
and at months 1, 3, 6, 9, and 12. RIjM(
P
This fact suggests that a new concept [`nY2[A$
This was accomplished by taking ... * nCx[
The preparatory stage is very time consuming process. tcOnM w
Test are performed for validity, completeness, and compatibility N-QCfDao
There is little hope of achieving successful ... gZ` DT
There has been an increasing awareness of the potential of using most ..so far made have wd+O5Lr.R
not taken this approach, with the exception of 4
z0L ke
Only a few studies can be found. !Uy>eji}
It is a very tedious process to go through +c?ie4
It is only when .. has been completed that .. may be effected r*Z_+a8
The entire interpretation process is conducted in one's head. ]!aUT&
These approaches are sometimes very tedious. qTy v.#{y
Several techniques can be used 2sun=3qb
A polynomial parametric model can be written as [the following]/[follows]: b1)\Zi
A xx model is constructed/formulated using xx. f4
O]`U
A xx model represents an xx by its xx. hBy*09Sv
A process decision model captures the logic essential to ]-Lruq#
From the equation above, xx is equal to the summation of xx times the ... LG6I_[
21 |^!Vo
&T
The validity of a xx model can be checked using Euler's formula. +7Rt{C,
Given a model, one can mathematically determine whether ... or ... =mLeMk/7 w
Equations for xx need to be derived and implemented in the system. oG\>--
A number of heuristic rules have been developed for S}mm\<=1
Optimum .. techniques can be made more reliable by ... so that eI?HwP{m
An algorithm based on the characteristic ... is used to determine C*Xik9n
Euler's formula states the following: z L8J`W
The completed model should agree with the formula. q;No"_aAd
For manufacturing purposes, a detailed and precise model of the object is necessary -)>(8 f
Engineering design models are very well defined; therefore, 1yJ75/
To keep the domain narrow enough to be implementable, yet wide enough to be useful. ,z oB0([
Point of View OQ,}/
from an implementation standpoint, <OIUyZS
From the point of view of this application, Fr2kbQTg;
From this point of view, Zadeh suggested an inference rule named xxx (CRI for short). -EVs@:3]j
Information is the meaningful interpretation and correlation of some aggregation of data n_1jHJo
in order to allow one to make decisions. OelU
D/[$
From a practical point of view, the computational aspects of an FLC require a 2'Y{FY_Z
simplification of the fuzzy control algorithm. 7oD
y7nV4
The use of a hammer to insert screws, although partly effective, tends to distort, destroy, Y}S.37|+^
and generally defeat the purpose of using a screw [Kusiak AI Implications for CIM 9@Q&B+!
p.129] jmSt?M0.xV
Statistical analysis Etw~*
Data were analyzed by one-way analysis of variance comparing the three conditions 4Q@\h=r
(sham operation, ischemic AKI, and bilateral nephrectomy) at each time point. If `|&\e_"DE
significant F-statistic from analysis of variance existed, this test was followed by Dunnett Bj; [
post hoc multiple comparison procedure with sham operation as the control group. For all /9P^{OZ;y
other comparisons, Student's t-test was used. A P-value of <0.05 was considered as AjG)1
statistically significant. YXgWH'i~
Values are expressed as means s.e.m. and significance was evaluated by Mann–Whitney u
IAZo;
U test using GraphPad Prism, version 4.0 software (GraphPad Software Inc., San Diego, vf6`s\6
CA, USA). y2U/$%B)G
All values are expressed as means s.d. Statistical significance (defined as P<0.05) was EK^JLvyT
evaluated using analysis of variance and Bonferroni t-tests, and the two-tailed Pearson's P`7ojXy
test, where appropriate. BQ).`f";d
Data are expressed as mean s.d., median and interquartile range, or frequencies, as ^g}L`9fL
appropriate. Variables who deviated from the normal distribution (positively skewed) 8b-7]%
were log-transformed (log10) before the correlation study. 0Wjd-rzc,
Data are represented as means s.e.m. Student's t-test and multiple comparisons with t-test sq6>DuBZz
post hoc analysis of variance were used as indicated below, for the comparison of f$G
r`d
22 r;c' NqP
morphological, immunohistological and functional parameters. Statistical significance ,?C|.5
was set at P<0.05. &\JK%X.Jlt
The primary efficacy parameter was defined as change in urinary protein excretion from FX{~"
baseline (week 0) to 12 months after treatment. The 12-month changes were tested =AWX
+znP
against zero using the paired t-test. Secondary end points included 6-month changes in es.jh
protein; the number with PR or CR at 6 or 12 months; and changes in glomerular kJHr&=VO~
filtration rate (GFR), serum albumin, and lipid profiles. Study sample size was based on }9{dR4hD
the desire to have 80% power to detect a drop in urinary protein of at least 2.0 g day-1. &mX5&e
Assuming a two-sided hypothesis test performed at a significance level of 0.05 and an s.d. *>EI2HX
of urinary protein change of 2.5 g, it was determined that 15 patients were required.2 6
"BtfQ")
Definition of remission status is according to the criteria established by Cattran et al.30 %M|Z}2qv
CR was defined as proteinuria <0.3 g per 24 h, PR as proteinuria 3 g per 24 h, and a aQz|!8Is
>50% reduction in peak proteinuria and non-response as <50% reduction in peak
s/r5,IFR
proteinuria. Any patient reaching a CR or PR was considered a treatment success 3$GY,B
The statistical significance of differences for the mean values of cytokine concentration vF5wA-3&t
and T cell proliferation was determined with Student's t-test. Differences with a P value SQ1M4:hP
of less than 0.05 were considered significant. #Tr;JAzVjG
Statistical significance was determined by Student's t-test. Data with a P value of less xI=}z
than 0.05 were considered significant. 1{SrHdD=
In vitro and in vivo experiments were analyzed by the two-tailed Student's t-test, with P 1Z^`l6|2
values less than 0.05 considered statistically significant. Tns?mQ
The significance of differences among groups was determined by Student's t-test and jF%)Bhn(
one-way analysis of variance (SigmaStat software; Jandel). -X"5G
Comparisons ^CowJ(y(
As well, the pros and cons of these representations from a process planning point of view @-)<|orU4
will be discussed. ,!o\),N
The method of using xx to implement xx described by Zadeh (1973) appeared more %NF<bEV
suitable `jD8(}_
As discussed [in the previous section]/[preciously], R&u)=~O\5
Relation -XS+Uv
We can not invert F' directly because it defines a many-to-one mapping. #~bU}[
{
The relationships appear very complicate l _kg3e4
Lifting tasks involve complex and imprecise relationship between the task variables and 9TYw@o5V
the human operator's characteristics. VfL]O 8P>
These methods are based on the relationship between ... and ... lfU"SSQ
The fundamental concept of a fuzzy rating language is that we can establish a relationship ZI2K-z'e
among terms such as high, medium, and low, and then modify these relationships. %5Q7 #xU
This article will thus mention the latter as well as the former. ^]#Ptoz^(l
The former two bear a close relation to a fuzzy Cartesian product. W,9k0t
Importance _/QKWk&j
) h3U
Z|B0=
23 :,h=2a_ 8
The emphasis is on an implementation of a general approach to rule based decision
Pqr Ou
making. k=h/i8i2z
Consideration / Attention mFgb_Cd
Careful evaluation is necessary to ensure iGEQXIr3
Such a formulation does not change further considerations. :|3"H&FWK
Considerable attention has been paid to zn2"swhq\V
Attention should be paid to an important finding of this investigation. OUBgBr
Caution should be exercised in this process to avoid ... E5Snl#Gl\0
Primary consideration is given to ... components, though others can be accommodated k2_6<v
Z
After ... has been defined by ..., a carefully analysis is carried out/performed to determine Pq{YZMr
A number of factors such as ...need to be taken into consideration before making the &geO
Fe}R
appropriate decision. Ru9pb~K
It should be noted that EvH(Po h
It is important to point out that ...
(<>Sz(
These considerations have heightened interest in the possibility of providing ... WoClTb>F
We should stress the fundamental importance of the xx 5JU(@}Db
Results. ys.!S.k+
Advantages / Disadvantage X('Q;^`
One of the major advantages of this new measure of xx is that it can be applied to the =B o4yN
experimental study of My6]k?;}(
One advantage of using a .. is the ease of preparing it. ,h^;~|GT
It has a very fast decision making process 93Ci$#<y
All the algorithms involve mostly logical operations. e~2*>5\:
It can be easily and without additional cost implemented in a microprocessor;based -fSKJo#}|
environment. ++p&
x{
It can reduce the waste of designing from scratch. p-
(ADQS
The advantages of using a xx to represent xx are the following: bbPd&7
However, xx is not without its shortcomings. =|{,5="
In most cases, the xxx shows an improvement over the existing xxx. }&C!^v
o
Compared to the existing xx, the impacts of the xx are generally reduced by 5% to 9%. KbuGf$Bv
The "best case" results shows a savings of 6% to 9%. BI]ut|Qw
Most of the existing works based on xx approach can only recognize a xx . n`I
jG
Most of the above methods are computational expansive and limited to xx. 3_1Io+uXk
Some other advantages of xx are the following: '`goy%Wd
The problem is the limitation of this method to a limited domain of parts. ,w58n%)H
It proved limited in application because it demanded precision in system modeling that $b
71
was impossible in practice. ;Q,).@<C
There are advantages to be gained in the structuring of costs and benefits, the use of xx, sxQ ,x/O
The disadvantages of this method are also disadvantages of conventional xx approaches. vM3|Ti>a'
This combines the best features of both techniques v=Bh
A9[
24 A[,"jh
Hopefully, this tool can be as the reference framework of for developing a xx platform, 2'^OtM,
and helping the administration, marketing, and knowledge management activities in [6gHi.`p'
virtual communities. {8p?we3l1
Results om39;nk!}
An improvement on the result shown above can be made by based on the data provided ^d$e^cU
Recent work has identified F1[[
fH
Time-dependent changes of {kO:HhUg
Cyclosporine-induced cell death is triggered by a non-classical phosphoinositide 3-kinase kX .1#%Ex
and does not require ERK activation. m3!MHe~t
Much of the current work on `TR9GWU+B
Discussion of these theories is beyond the scope of this review "&o@%){]
Based on the information contained in this LK}eU,m=
The result can be categorized into nine classes (S 3kP5:F
The results are illustrated by an example @ y2Bq['
The experimental results for each xx time are reported in Table 2. ,r,;2,;6nd
From the results obtained so far, it seem that mpJ_VS`
Because of the inaccuracy of the ..., a conclusion cannot be drawn as YZu#0)
Although much effort has been made to., this reality is far from completion. KEB>}_[
The results indicate that the total benefits are higher than the total costs. U,}T ]J
Their results may then serve as guidelines for lower level models, less fuzzy and more \V\ET
detailed. 0# GwhB
Conclusion u<r('IW0
From the discussion, one may conclude that ...
GB Un" _J
The first indication that !Bbwl-e`
In summary, we have shown that ptfADG
These results suggested that O@&I.d$
Our findings have shown that ".Z+bi2l
Our observations have provided ozwqK oE
This study reveals following five main findings: $l"(tB7d
To our knowledge, this is the first immunohistochemical report of both PIM and HIFs in 'v)+S;oB
the diabetic kidney. }('QIvq2
As mentioned above, IMEoov-x
Moreover, we demonstrate that, OPh@H.)^
A number of studies have elaborated a body of knowledge about ZS;kCdL
There are findings to indicate that >qZRIDE5$
On a descriptive level, there is agreement that for M,lu)~H
One issue that became particularly evident from our study is aa!c>"g6
Our data are in agreement with the findings and models presented by previous m8`A~
publications, in particular those of Jones et al '1
*MiFxKq
According to our observations, ,^T2hY`
…was not evident from our data yV`vu/3K
25 _en 8hi@Z
It could therefore be concluded that 8i"fhN3?Y
our data illustrate that 5lbh
"m=
All these different aspects, as listed above, lead to jws(`mIf\
In fact, it has already been speculated that ~yg9ZM
It should be furthermore mentioned in this context that B dUyI_Ks:
To identify and verify the essential factors triggering developmental renin expression, it VI{1SIhfa
is of interest now to study the development of intrarenal renin expression in mice with =X):Zi
defined gene mutations, using the results of this study as a reference system. 48VsHqG
In conclusion, using several approaches, we demonstrated that FeJ5^Gh.
But we have not provided formal proof yet that VB%xV
In summary, our results suggest that
_L ].n)b
In conclusion, we identify "`wq:$R
A better understanding of such regulatory systems may yield novel therapeutic c@P,
approaches to glomerular diseases. &-9D.'WzP
Preliminary results of short-term studies (2 weeks) indicate that el<nY"
c
However, we believe that 6 s/O\A
However, despite our extensive measurements of these parameters, we found no )/Vr 5b@
relationship between response and initial proteinuria, time to B-cell recovery, the 1*"Uc!7.%
development of antibodies to rituximab, B-cell count in the kidney tissue, nor the degree hV%l}6yS&
of tubular interstitial damage present. A clear conclusion for efficacy, however, cannot be ),_bDI L+
obtained from an uncontrolled study, and definitive claims of efficacy should be reserved w>e
OERZa
for rigorous, prospective, controlled, and randomized trials with the use of rituximab and A@4Cfb@
that will also look for factors that may determine its efficacy in IMN. b]~M$y60q
It has become increasingly clear that TFhj]r^{
Form the above discussion, the conclusion can be reached that S70ERRk
The conclusions drawn are also valid 6eBQ9XV
In conclusion to this, it becomes obvious that the problem of xx lies not only in... sr\MQ?\fB
We have attempted to introduce some concepts associated with a theory of ` mi!"pm w
Considerable more work, hopefully, will be done in this area TCYnErqk
Employing the compositional rule of inference, the assessment of the xx compatibility in $t]DxMd
achieving prescribed xx projectiles in any level of the hierarchy is made possible. ^ZsME,
This paper has presented a theoretical and experimental study of the xx process and xx r&qD