18 |Ax~zk;
Jurkat cells cultured in calcium-free RPMI-1640 medium (Gibco BRL; number *_3+ DF
22300-107) containing calcium-free 10% FBS were triggered by anti-Fas IgM. The a{,EX[~b
treated cells were harvested at the indicated time points and incubated with Fluo-3-AM at VI[ikNpX
a final concentration of 1 micromolar for 30 min at 37°C (Scoltock et al., 2000). The A1>R8Zuhy
labeled cells (50,000 cells per treatment) were then analyzed by exciting the cells at 488 X{<j%PdC
nm and examining the fluorescence emission of Fluo 3 at 530 nm with a FACS Scan, eB^:+h#A_
(Becton Dickinson). A one micromolar concentration of LPA was used as a positive l^WFMeMD3a
control for Ca2+ induction. The data thus obtained was analyzed with the software Win Ga"$_DyM
MDI 2.8 and represented as contour plots.The effect of chelating intracellular calcium on 'C!b($Y
translocation of annexin I was studied by culturing Jurkat cells in the presence of 10 R/A40i
micromolar BAPTA-AM, with or without the addition of anti-Fas IgM. Cells were >/eQjp?:
harvested and fractionated as detailed above, and the S-100 fractions were assessed by n<y!@p^X
immunoblotting for presence or absence of annexin I. N@)tU;U3O
Mouse bone marrow transduction and transplantation. '$UlJDZ
Retrovirus-mediated transduction of mouse bone marrow cells was done by published +wgNuj0=*
methods49. Prestimulated low-density bone marrow cells were infected with high-titer 1j9 .Q;9
retrovirus supernatant on fibronectin-coated plates. Retrovirus supernatant was generated CXb)k.L
in the phoenix-gp cells with a mouse stem cell virus–based retroviral vector coexpressing jx+%X\zokA
EGFP and HA–RhoH as described50. EGFP+ sorted cells were transplanted by r,EIOcz:
intravenous injection into the sublethally irradiated (300 rads with a 137Cs irradiator) q_ykB8Ensa
Rag2-/- recipient mice. At 9 weeks after transplantation, thymus, peripheral blood, bone Y*dzoN.sW
marrow, spleen and lymph nodes from each recipient mouse were collected for analysis / F5g@ X&
of EGFP+ chimerism and hematopoietic lineage by flow cytometry. Expression of 3;>|*(cO
HA–RhoH and HA–RhoHF73F83 in EGFP+ sorted thymocytes of recipient mice was m("!
M~1
confirmed by immunoblot analysis. ZS4lb=)G
Determination of renal morphology #,0%g1
Kidney slices were postfixed in buffered 2% OsO4, dehydrated, and embedded in an tz6d
}$
Araldite-EM bed 812 mixture. Large sections were cut perpendicular to the renal capsule, +Y0Wiwr'
containing cortex, and medulla. Thin (1 m) sections were analyzed in a blinded manner (B}+uI{
for morphologic alterations, as previously detailed 9pb4!=g*
Patient population 2$jTj<.K
Patients included in the study met the following criteria: (1) biopsy-proven IMN; (2) "nf.kj:>
creatinine clearance 30 ml per min per 1.73 m2; and (3) persistent proteinuria >5 g per o2He}t2o
24 h despite treatment with an HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A) NgaX&m`
reductase inhibitor, an ACEi, and/or ARB at maximal tolerated dose for at least 4 months. AL>*Vj2h/n
The Mayo Institutional Review Board and the Research Ethical Board, University Health
A<C`JN}
Network, University of Toronto approved the study protocol. All patients gave written X@*$3z#Z
informed consent. Patients who had been on treatment with prednisone, cyclosporine, or s"*zyLUUo
19 3U!\5Nsby
mycophenolic mofetil within the last 4 months or alkylating agents within the last 6 !i&
^H,
months were not included in the study. Patients with active infection, diabetes, or a ek1Y
aE
secondary cause of MN (for example, hepatitis B, systemic lupus erythematosus (SLE), >%h_ R:
medications, malignancies) were also excluded. R+
lwOVX
Treatment -n?}L#4%8
At enrollment, a low-sodium (<4 g day-1) and low-protein (0.8 g per kg per day of Dt0S"`^=k
high-quality protein) diet was recommended and patients were encouraged to maintain &B1d+.+
the same diet throughout the duration of the study. All patients received a similar E5w.wx
conservative treatment regimen that included loop diuretics to control edema, an kL 6f^MoL
HMG-CoA reductase inhibitor, and an ACEi combined with an ARB if tolerated. lX/6u
E_%
-Blockers and non-dihydropyridine calcium channel blockers, in that order, were added jhr{JApbJv
when required to control systolic blood pressures to <135 mm Hg in >75% of the *z!!zRh3x
readings. Patients who after a minimum of 4 months of conservative therapy and 8l-+
4~mH
maximized Ang II blockade had proteinuria >5 g per 24 h received two i.v. infusions of (D]l/akP
rituximab at a dose of 1000 mg on days 1 and 15. To minimize infusion reactions, {}H5%W
patients were premedicated with acetaminophen (1000 mg) and diphenhydramine wXIsc;
hydrochloride (50 mg) orally. In addition, methylprednisolone (100 mg, i.v.) was given h?jy'>T?b2
prior to the first rituximab infusion. B-cell depletion was defined as CD19+ count `c ~Va/Yi
<5 cells per l at any time and B-cell recovery was defined as CD19+ cell count rl qn39
>15 cells per l. Patients treated with rituximab, who at month 6 had proteinuria >3 g per *lRP ZN
24 h and in whom CD19+ B-cell counts had increased to >15 cells per l, received a L
']EYK5
second course of rituximab treatment following the same protocol described above.
uh*b[`e
Follow-up Qctm"
g|
In all patients, clinical and laboratory parameters including complete blood counts, p:U9#(v)
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum Gnop
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry Xm`jD'G
and at months 1, 3, 6, 9, and 12. Creatinine clearance and protein and creatinine excretion h~)oiT2v
in the urine were assessed by performing two consecutive 24-h urine collections at each Kf2Ob1
time point. Data were considered accurate when urinary creatinine excretion was fEf",{I
consistent with a complete 24 h collection. The mean of the two measurements was p8o
~
considered for the analysis. The presence of HACAs was evaluated at baseline and at %B5.zs]Of
months 3, 6, 9, and 12. %p6"Sg*
Method / Approach / Study/ Technique ,rVm81-2
A discussion is presented of a problem-solving system Sr Z\]
To improve the efficiency of the method, the following approach may be applied. c-.t8X,5(~
In order to an investigation was made to find the causes of the Vui5Z K
Although large collections of rules and equations have been complied, none are generally [ R1S+i
accepted Tu2
BQ4\[
20 ]BGWJ A5
This approach will be explained and discussed thoroughly in the body of the report. S^'?sfq
This can be accomplished by dmI,+hHtL
This algorithm to compute the total cost can be described step by step as follows: 0j\?zt?
The above preliminary analysis has provided important information V1 T?T9m
Various methods have been proposed for selecting an optimum... S e/VOzzg
These concepts have been applied to NKf][!bi
On the basis of the concept mentioned above, i lk\&J~I
This can be achieved by ?JO x9;`
In addition, tissues were stained for infiltrating lymphocytes (CD20 and CD3), and the sN0S~}F+
amount of interstitial fibrosis was quantified by histomorphometry. Formalin-fixed, rM<|<6(L
paraffin-embedded sections were cut onto coated glass slides. Following heat-induced #
le<R
antigen retrieval, sections were incubated at 20°C overnight with either anti-CD20 >ly`1t1
primary antibody or anti-CD3 primary antibody, both at 1:1000 dilution (Dako, Canada {E`[`Kf
Inc, Mississauga, Ontario, Canada). After rinsing all sections, pretreatment with 3% gjB36R
hydrogen peroxide was performed to prevent endogenous peroxidase activation. Sections <+@?V$&
were incubated with a secondary rabbit anti-mouse antibody linked with avidin–biotin '7AlE!7%
complex. Sections were counterstained with hematoxylin and examined by light PhL }V|W>
microscopy. 17g\XC@ Cl
The HACA assay is a proprietary bridging enzyme-linked immunosorbent assay &TYTeJ]
performed at Genentech Inc. that measures the antibody response to rituximab in human B@g 0QgA
serum samples. ^]:w5\DG
In all patients, clinical and laboratory parameters including complete blood counts, `F5iZWW1
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum J%`-K"NB
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry
x~p8Mcv
and at months 1, 3, 6, 9, and 12. G?$|aQ0j
This fact suggests that a new concept
|>o]+ V
This was accomplished by taking ... h?E[28QB
The preparatory stage is very time consuming process. iIWz\FM
Test are performed for validity, completeness, and compatibility :{@&5KQ8)
There is little hope of achieving successful ... xL8r'gV@
There has been an increasing awareness of the potential of using most ..so far made have Sc03vfmo"N
not taken this approach, with the exception of V|GH4DT=
Only a few studies can be found. qW*JB4`?a
It is a very tedious process to go through
LVNq@,s
It is only when .. has been completed that .. may be effected S9/oBxGN
The entire interpretation process is conducted in one's head. _GEt:=DAP#
These approaches are sometimes very tedious. R
gY-fc0
Several techniques can be used xCU^4DO3p
A polynomial parametric model can be written as [the following]/[follows]: ):"Z7~j=
A xx model is constructed/formulated using xx. 4q sIJJ[.
A xx model represents an xx by its xx. y+b4s
Ff
A process decision model captures the logic essential to _BGw)Z 6
From the equation above, xx is equal to the summation of xx times the ... %Jy0?W N
21
B-Bgk
The validity of a xx model can be checked using Euler's formula. \Tq !(]o^
Given a model, one can mathematically determine whether ... or ... )I7~<$w
Equations for xx need to be derived and implemented in the system. \Tf[% Kt x
A number of heuristic rules have been developed for 6mcxp+lm|
Optimum .. techniques can be made more reliable by ... so that o,o,(sII
An algorithm based on the characteristic ... is used to determine hP1}Do
Euler's formula states the following: )1 ]P4
The completed model should agree with the formula. 6)<g%bH!
For manufacturing purposes, a detailed and precise model of the object is necessary oU/CXz?H
Engineering design models are very well defined; therefore, q\+khy,k
To keep the domain narrow enough to be implementable, yet wide enough to be useful. <1")JDW
Point of View 5f5
bhBZ<
from an implementation standpoint, )qXl8H I
From the point of view of this application, 80qSPitj
From this point of view, Zadeh suggested an inference rule named xxx (CRI for short). mOiA}BGw
Information is the meaningful interpretation and correlation of some aggregation of data h#_KO-#.[
in order to allow one to make decisions. M0Eq
7:Ba
From a practical point of view, the computational aspects of an FLC require a !~X[qT
simplification of the fuzzy control algorithm. "S8JHHx
The use of a hammer to insert screws, although partly effective, tends to distort, destroy, y b 7
and generally defeat the purpose of using a screw [Kusiak AI Implications for CIM #<_gY
p.129] LjZvWts?
Statistical analysis TVaD',5_V%
Data were analyzed by one-way analysis of variance comparing the three conditions +7yirp~`K
(sham operation, ischemic AKI, and bilateral nephrectomy) at each time point. If Xx.4K>j+j
significant F-statistic from analysis of variance existed, this test was followed by Dunnett )8BGN'jyi
post hoc multiple comparison procedure with sham operation as the control group. For all um/2.Sn>
other comparisons, Student's t-test was used. A P-value of <0.05 was considered as BGk>:Z`
statistically significant. =m<; Jx5
Values are expressed as means s.e.m. and significance was evaluated by Mann–Whitney 9VqE:c /
U test using GraphPad Prism, version 4.0 software (GraphPad Software Inc., San Diego, XwI~ 0
CA, USA). 6??o(ziK$
All values are expressed as means s.d. Statistical significance (defined as P<0.05) was m8Wv46%
evaluated using analysis of variance and Bonferroni t-tests, and the two-tailed Pearson's /g|H?F0
test, where appropriate. +ZuT\P&kR5
Data are expressed as mean s.d., median and interquartile range, or frequencies, as :0G_n\
appropriate. Variables who deviated from the normal distribution (positively skewed) $-HP5Kj(k-
were log-transformed (log10) before the correlation study. t r)[6o#
Data are represented as means s.e.m. Student's t-test and multiple comparisons with t-test 4r'QP .h
post hoc analysis of variance were used as indicated below, for the comparison of "6[a%f#Q
22 B8XW+U
morphological, immunohistological and functional parameters. Statistical significance 0F"W~OQ6
was set at P<0.05. <>gX'te
The primary efficacy parameter was defined as change in urinary protein excretion from O=,[u?
baseline (week 0) to 12 months after treatment. The 12-month changes were tested jkTh)Bm|'
against zero using the paired t-test. Secondary end points included 6-month changes in 79DzrLu
protein; the number with PR or CR at 6 or 12 months; and changes in glomerular =r w60B
filtration rate (GFR), serum albumin, and lipid profiles. Study sample size was based on |2i=oX(r|
the desire to have 80% power to detect a drop in urinary protein of at least 2.0 g day-1. W^a-K
Assuming a two-sided hypothesis test performed at a significance level of 0.05 and an s.d. HDmjt+3&n
of urinary protein change of 2.5 g, it was determined that 15 patients were required.2 K;:_UJ>t
Definition of remission status is according to the criteria established by Cattran et al.30 [uJfmr EH
CR was defined as proteinuria <0.3 g per 24 h, PR as proteinuria 3 g per 24 h, and a {;z{U;j
>50% reduction in peak proteinuria and non-response as <50% reduction in peak Ri<7!Y?l
proteinuria. Any patient reaching a CR or PR was considered a treatment success jTR>H bh
The statistical significance of differences for the mean values of cytokine concentration 9.:]eL
and T cell proliferation was determined with Student's t-test. Differences with a P value :.k1="H~@
of less than 0.05 were considered significant. 3"*tP+H
Statistical significance was determined by Student's t-test. Data with a P value of less ?S*Cvr+=4
than 0.05 were considered significant. n7|8`?R^
In vitro and in vivo experiments were analyzed by the two-tailed Student's t-test, with P &["s/!O1 R
values less than 0.05 considered statistically significant. m1 tYDZ"i
The significance of differences among groups was determined by Student's t-test and 6`c5\G+
one-way analysis of variance (SigmaStat software; Jandel). kFRl+,bi~
Comparisons s6DmZ^Y%
As well, the pros and cons of these representations from a process planning point of view xJ$/#UdP
will be discussed. %Tv2op
The method of using xx to implement xx described by Zadeh (1973) appeared more <:n!qQS6
suitable &x":
As discussed [in the previous section]/[preciously], 5[[
mS
Relation s|`Z V^R
We can not invert F' directly because it defines a many-to-one mapping. +HXR ))X
The relationships appear very complicate qxQuXF>:#
Lifting tasks involve complex and imprecise relationship between the task variables and n@JZ 2K4
the human operator's characteristics. hZ[(Ik]*Zd
These methods are based on the relationship between ... and ... zao=}j?
The fundamental concept of a fuzzy rating language is that we can establish a relationship A_4.>g
among terms such as high, medium, and low, and then modify these relationships. *el~sor;S
This article will thus mention the latter as well as the former. ^pysoaZCT_
The former two bear a close relation to a fuzzy Cartesian product. 6CV9ewr
Importance 7'g{:dzS*3
) +_T`tmQ
23 *U mWcFoF
The emphasis is on an implementation of a general approach to rule based decision q` 0wG3
making. BN~g
k~t_
Consideration / Attention 8#{DBWU
Careful evaluation is necessary to ensure Xl %ax!/
Such a formulation does not change further considerations. fP KFU
Considerable attention has been paid to 7C>5Xyy
J
Attention should be paid to an important finding of this investigation. u85dG7
Caution should be exercised in this process to avoid ... 6ec#3~ Y]
Primary consideration is given to ... components, though others can be accommodated v,d
bto0
After ... has been defined by ..., a carefully analysis is carried out/performed to determine KNG7$icG
A number of factors such as ...need to be taken into consideration before making the 61~7 L^882
appropriate decision. ]ft}fU5C1
It should be noted that YHOo6syk
It is important to point out that ... 'Z8=y[l
These considerations have heightened interest in the possibility of providing ... UbEK2&q/8
We should stress the fundamental importance of the xx hd1aNaF-
Results. +pY--5t
Advantages / Disadvantage a{@gzB
One of the major advantages of this new measure of xx is that it can be applied to the $]<wQH/?_
experimental study of +GvPJI
One advantage of using a .. is the ease of preparing it. #GuN.`__n,
It has a very fast decision making process I&9Itn p$
All the algorithms involve mostly logical operations. fZavZ\qU
It can be easily and without additional cost implemented in a microprocessor;based cd(YH! 3
environment. 1d 1
~`B
It can reduce the waste of designing from scratch. rHp2I6.0a
The advantages of using a xx to represent xx are the following: 0fog/c#q(
However, xx is not without its shortcomings. ;-^8lWt
In most cases, the xxx shows an improvement over the existing xxx. KA? J:
Compared to the existing xx, the impacts of the xx are generally reduced by 5% to 9%. }u]7 x:lh
The "best case" results shows a savings of 6% to 9%. l.xKv$uOGR
Most of the existing works based on xx approach can only recognize a xx . _P9*78
Most of the above methods are computational expansive and limited to xx. Vr:`?V9Q2(
Some other advantages of xx are the following: $'n?V=4
The problem is the limitation of this method to a limited domain of parts. 9~{,Hj1xE
It proved limited in application because it demanded precision in system modeling that ZkryoIQ%=
was impossible in practice. ^#!\VGnL
There are advantages to be gained in the structuring of costs and benefits, the use of xx, .dsB\C
The disadvantages of this method are also disadvantages of conventional xx approaches. j@R"AP}
This combines the best features of both techniques CrB4%W:{
24 TPn#cIPG
Hopefully, this tool can be as the reference framework of for developing a xx platform, yRR[M@Y
and helping the administration, marketing, and knowledge management activities in U%n>(!d
virtual communities. jAdZS\?w
Results !j
/54,
An improvement on the result shown above can be made by based on the data provided >P ~j@Lv
Recent work has identified uann'ho?q
Time-dependent changes of [r1dgwh8
Cyclosporine-induced cell death is triggered by a non-classical phosphoinositide 3-kinase 1&RB=7.h
and does not require ERK activation. =E62N7_`=
Much of the current work on 48Jt1^
Discussion of these theories is beyond the scope of this review &