18 NoO>CjeFb
Jurkat cells cultured in calcium-free RPMI-1640 medium (Gibco BRL; number <Q/^[
22300-107) containing calcium-free 10% FBS were triggered by anti-Fas IgM. The m(q6Xe:Vc
treated cells were harvested at the indicated time points and incubated with Fluo-3-AM at 3H%R`ha
a final concentration of 1 micromolar for 30 min at 37°C (Scoltock et al., 2000). The k^Zpb&`Hx
labeled cells (50,000 cells per treatment) were then analyzed by exciting the cells at 488 O_K@\<;~
nm and examining the fluorescence emission of Fluo 3 at 530 nm with a FACS Scan, \#1!qeF
(Becton Dickinson). A one micromolar concentration of LPA was used as a positive bxyEn'vNvQ
control for Ca2+ induction. The data thus obtained was analyzed with the software Win Wv_5sPqLW
MDI 2.8 and represented as contour plots.The effect of chelating intracellular calcium on >9a%"<(2#
translocation of annexin I was studied by culturing Jurkat cells in the presence of 10 2FD[D`n]f
micromolar BAPTA-AM, with or without the addition of anti-Fas IgM. Cells were 9Re605xQ6
harvested and fractionated as detailed above, and the S-100 fractions were assessed by 6#T?g7\pyR
immunoblotting for presence or absence of annexin I. ?=jmyDXH!
Mouse bone marrow transduction and transplantation. +cvz
Retrovirus-mediated transduction of mouse bone marrow cells was done by published |2CW!is
methods49. Prestimulated low-density bone marrow cells were infected with high-titer aQCbR
S6
retrovirus supernatant on fibronectin-coated plates. Retrovirus supernatant was generated :>nk63V (
in the phoenix-gp cells with a mouse stem cell virus–based retroviral vector coexpressing Fly@"W4a
EGFP and HA–RhoH as described50. EGFP+ sorted cells were transplanted by _Jk-nZgn
intravenous injection into the sublethally irradiated (300 rads with a 137Cs irradiator) CEc&
G
Rag2-/- recipient mice. At 9 weeks after transplantation, thymus, peripheral blood, bone UYW%%5p?
marrow, spleen and lymph nodes from each recipient mouse were collected for analysis Vyj>&"28
of EGFP+ chimerism and hematopoietic lineage by flow cytometry. Expression of -kES]P?2
HA–RhoH and HA–RhoHF73F83 in EGFP+ sorted thymocytes of recipient mice was
$8_t.~q
confirmed by immunoblot analysis. n]ar\f
Determination of renal morphology ZgtW
Kidney slices were postfixed in buffered 2% OsO4, dehydrated, and embedded in an T=VVK6Lc:
Araldite-EM bed 812 mixture. Large sections were cut perpendicular to the renal capsule, [Qy]henK
containing cortex, and medulla. Thin (1 m) sections were analyzed in a blinded manner JQ+4 SomK
for morphologic alterations, as previously detailed ETH`.~
%
Patient population A&$!s)8z
Patients included in the study met the following criteria: (1) biopsy-proven IMN; (2) s[nOB0
creatinine clearance 30 ml per min per 1.73 m2; and (3) persistent proteinuria >5 g per Vt
n$*ML
24 h despite treatment with an HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A) (d^pYPr{
reductase inhibitor, an ACEi, and/or ARB at maximal tolerated dose for at least 4 months. 7Yk6C5C
The Mayo Institutional Review Board and the Research Ethical Board, University Health s7FqE>#c0
Network, University of Toronto approved the study protocol. All patients gave written %E1~I\n:F
informed consent. Patients who had been on treatment with prednisone, cyclosporine, or IZJV6clM
19 Q
;m
.m2
mycophenolic mofetil within the last 4 months or alkylating agents within the last 6 WRVKh
months were not included in the study. Patients with active infection, diabetes, or a L}>9@?;GW
secondary cause of MN (for example, hepatitis B, systemic lupus erythematosus (SLE), p*Q"<@n
medications, malignancies) were also excluded. b\=0[kBQw
Treatment EoQ.d|:g
At enrollment, a low-sodium (<4 g day-1) and low-protein (0.8 g per kg per day of Pl1:d{"d
high-quality protein) diet was recommended and patients were encouraged to maintain gQ?>%t]
the same diet throughout the duration of the study. All patients received a similar q n =6>wP
conservative treatment regimen that included loop diuretics to control edema, an
O(&EnNm[2
HMG-CoA reductase inhibitor, and an ACEi combined with an ARB if tolerated. N:L<ySJ7
-Blockers and non-dihydropyridine calcium channel blockers, in that order, were added WpTC,~-
when required to control systolic blood pressures to <135 mm Hg in >75% of the . s?
''/(
readings. Patients who after a minimum of 4 months of conservative therapy and a Mv
maximized Ang II blockade had proteinuria >5 g per 24 h received two i.v. infusions of cB -XmX/
rituximab at a dose of 1000 mg on days 1 and 15. To minimize infusion reactions, Hd;>k$B
patients were premedicated with acetaminophen (1000 mg) and diphenhydramine +"'h?7'C
hydrochloride (50 mg) orally. In addition, methylprednisolone (100 mg, i.v.) was given Dm 'Q&
prior to the first rituximab infusion. B-cell depletion was defined as CD19+ count j{p0yuZ)<
<5 cells per l at any time and B-cell recovery was defined as CD19+ cell count OEB_LI'
>15 cells per l. Patients treated with rituximab, who at month 6 had proteinuria >3 g per AqN(htGvx
24 h and in whom CD19+ B-cell counts had increased to >15 cells per l, received a .YS[Md{
second course of rituximab treatment following the same protocol described above. /x$ jd)C
Follow-up x!I7vs~~zW
In all patients, clinical and laboratory parameters including complete blood counts, I9Z8]Q+2"
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum w'
L;`k;Q
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry u;=("S{"0
and at months 1, 3, 6, 9, and 12. Creatinine clearance and protein and creatinine excretion ^GRd;v=-@
in the urine were assessed by performing two consecutive 24-h urine collections at each {Ziq~{W_
time point. Data were considered accurate when urinary creatinine excretion was .Ta$@sP h}
consistent with a complete 24 h collection. The mean of the two measurements was 4{fi=BA
considered for the analysis. The presence of HACAs was evaluated at baseline and at VLQf
uh;
months 3, 6, 9, and 12. ^]aD
LjD
Method / Approach / Study/ Technique ?5/Sa
A discussion is presented of a problem-solving system 2H]&3kM3X
To improve the efficiency of the method, the following approach may be applied. Yt|{l
In order to an investigation was made to find the causes of the `^_.E:f
Although large collections of rules and equations have been complied, none are generally [k1N `K(M
accepted c&'T By
20 ->93.sge
This approach will be explained and discussed thoroughly in the body of the report. \f
This can be accomplished by t3bN
PK^
This algorithm to compute the total cost can be described step by step as follows: $9hOWti
The above preliminary analysis has provided important information "J[Cr m
Various methods have been proposed for selecting an optimum... (a)d7y.oo
These concepts have been applied to RYvS,hf6z
On the basis of the concept mentioned above, ri\r%x
This can be achieved by U)c,ZxE
In addition, tissues were stained for infiltrating lymphocytes (CD20 and CD3), and the F;MFw2G
amount of interstitial fibrosis was quantified by histomorphometry. Formalin-fixed, iCw~4KG
paraffin-embedded sections were cut onto coated glass slides. Following heat-induced %Xp}d5-
antigen retrieval, sections were incubated at 20°C overnight with either anti-CD20 ,T1t`
primary antibody or anti-CD3 primary antibody, both at 1:1000 dilution (Dako, Canada wcV~z:&^5
Inc, Mississauga, Ontario, Canada). After rinsing all sections, pretreatment with 3% AZtZa'hbkQ
hydrogen peroxide was performed to prevent endogenous peroxidase activation. Sections Iy`Zh@"~
were incubated with a secondary rabbit anti-mouse antibody linked with avidin–biotin dq|z;,`
complex. Sections were counterstained with hematoxylin and examined by light v
?Y9z!M
microscopy. mtJI#P
The HACA assay is a proprietary bridging enzyme-linked immunosorbent assay w"A'uFXLc
performed at Genentech Inc. that measures the antibody response to rituximab in human v( (fRX.`
serum samples. IURi90Ir
In all patients, clinical and laboratory parameters including complete blood counts, Af *^u|#
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum @gs
Kb*,
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry M
[Ls:\1a
and at months 1, 3, 6, 9, and 12. G~^Pkl3%T
This fact suggests that a new concept 3pq&TYQU
This was accomplished by taking ... =;c? 6{<1
The preparatory stage is very time consuming process. 5H?`a7q N
Test are performed for validity, completeness, and compatibility >dZ x+7
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There has been an increasing awareness of the potential of using most ..so far made have kqv>rA3
not taken this approach, with the exception of j1<@*W&b
Only a few studies can be found. ^9"|tWf6O
It is a very tedious process to go through `@=}5 9+|
It is only when .. has been completed that .. may be effected 1
!OQxY}f
The entire interpretation process is conducted in one's head. hIPDJ1a
These approaches are sometimes very tedious. a<c % Xy/
Several techniques can be used fl+
[(x<
A polynomial parametric model can be written as [the following]/[follows]: {"'W!WTb
A xx model is constructed/formulated using xx. Q_LPLmM
A xx model represents an xx by its xx. 3}n=o d=
A process decision model captures the logic essential to ~D<o}ItRF
From the equation above, xx is equal to the summation of xx times the ... Vx-HW;,
21 wdoA>a?q
The validity of a xx model can be checked using Euler's formula. xzIs,i}U
Given a model, one can mathematically determine whether ... or ... <k}>eGn
Equations for xx need to be derived and implemented in the system. Z4bN|\I
A number of heuristic rules have been developed for d/Fjs0pt
Optimum .. techniques can be made more reliable by ... so that ~F>oNbJIv
An algorithm based on the characteristic ... is used to determine 0O"GI33Mg
Euler's formula states the following: e+4p__TmZ
The completed model should agree with the formula. C[? itk!
For manufacturing purposes, a detailed and precise model of the object is necessary C6CX{IA]
Engineering design models are very well defined; therefore, Yca9G?^\v
To keep the domain narrow enough to be implementable, yet wide enough to be useful. {HvR24#
Point of View x-CYG?-x
from an implementation standpoint, 6i%LM`8GEk
From the point of view of this application, ~ $!eB/6ty
From this point of view, Zadeh suggested an inference rule named xxx (CRI for short). Fov/?
:f$
Information is the meaningful interpretation and correlation of some aggregation of data 1:q55!b
in order to allow one to make decisions. noh3mi
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simplification of the fuzzy control algorithm. q;tsA"l
The use of a hammer to insert screws, although partly effective, tends to distort, destroy, OQW#BBet@
and generally defeat the purpose of using a screw [Kusiak AI Implications for CIM
uSM4:!8
p.129] \*!g0C8 o
Statistical analysis a5L#c=
Data were analyzed by one-way analysis of variance comparing the three conditions )|uPCZdLZ
(sham operation, ischemic AKI, and bilateral nephrectomy) at each time point. If |C7GI[P
significant F-statistic from analysis of variance existed, this test was followed by Dunnett JS}W4 N
post hoc multiple comparison procedure with sham operation as the control group. For all u)R>ozER
other comparisons, Student's t-test was used. A P-value of <0.05 was considered as 3`IDm5
statistically significant. >OZ+k(saL
Values are expressed as means s.e.m. and significance was evaluated by Mann–Whitney <MA!?7Z|
U test using GraphPad Prism, version 4.0 software (GraphPad Software Inc., San Diego, H<X4R
CA, USA). #}:VZ2Z
All values are expressed as means s.d. Statistical significance (defined as P<0.05) was ~9h6"0K!
evaluated using analysis of variance and Bonferroni t-tests, and the two-tailed Pearson's NTs< ;ED
test, where appropriate. <)?H98S
Data are expressed as mean s.d., median and interquartile range, or frequencies, as 2s?j5 Sd
appropriate. Variables who deviated from the normal distribution (positively skewed) 5(u7b
were log-transformed (log10) before the correlation study. K)l{3\9l|
Data are represented as means s.e.m. Student's t-test and multiple comparisons with t-test Qn[4 &nUD
post hoc analysis of variance were used as indicated below, for the comparison of p
Ic;9
22 o=q
N+-N
morphological, immunohistological and functional parameters. Statistical significance ;# {XNq<1
was set at P<0.05. JjyQ
The primary efficacy parameter was defined as change in urinary protein excretion from A9p$5jt7
baseline (week 0) to 12 months after treatment. The 12-month changes were tested _o' jy^
against zero using the paired t-test. Secondary end points included 6-month changes in Fl(+c0|kT
protein; the number with PR or CR at 6 or 12 months; and changes in glomerular j%]i#iqF
filtration rate (GFR), serum albumin, and lipid profiles. Study sample size was based on aNv6 "
the desire to have 80% power to detect a drop in urinary protein of at least 2.0 g day-1. !COaPrg
Assuming a two-sided hypothesis test performed at a significance level of 0.05 and an s.d. \>23_d0
of urinary protein change of 2.5 g, it was determined that 15 patients were required.2 nU[ROy5
Definition of remission status is according to the criteria established by Cattran et al.30 1p+2*c
CR was defined as proteinuria <0.3 g per 24 h, PR as proteinuria 3 g per 24 h, and a $-5iwZ
>50% reduction in peak proteinuria and non-response as <50% reduction in peak W%Br%VQJ
proteinuria. Any patient reaching a CR or PR was considered a treatment success wvx
N6
The statistical significance of differences for the mean values of cytokine concentration vb5tyY0c
and T cell proliferation was determined with Student's t-test. Differences with a P value v|t^th,
of less than 0.05 were considered significant. Il~ph9{JH
Statistical significance was determined by Student's t-test. Data with a P value of less 6LNm>O
than 0.05 were considered significant. YOP=gvZq
In vitro and in vivo experiments were analyzed by the two-tailed Student's t-test, with P W*`2lf
values less than 0.05 considered statistically significant. SZ1+h TY7d
The significance of differences among groups was determined by Student's t-test and 9p>3k&S
one-way analysis of variance (SigmaStat software; Jandel). Z^>3}\_v
Comparisons jW?.>(
As well, the pros and cons of these representations from a process planning point of view ^[\F uSL
will be discussed. Ax=Rb
B"
The method of using xx to implement xx described by Zadeh (1973) appeared more _9\ayR>d
suitable DH)@8)C
As discussed [in the previous section]/[preciously], W O|2x0K
Relation @xSS`&b
We can not invert F' directly because it defines a many-to-one mapping. k2k/v[60
The relationships appear very complicate \qNj?;B
Lifting tasks involve complex and imprecise relationship between the task variables and l')?w]|
the human operator's characteristics. vxk1RL*Xu
These methods are based on the relationship between ... and ... (!W:-|[K\
The fundamental concept of a fuzzy rating language is that we can establish a relationship ,%v
among terms such as high, medium, and low, and then modify these relationships. 9_5Fl,u
z
This article will thus mention the latter as well as the former. S=eY`,'#R
The former two bear a close relation to a fuzzy Cartesian product. N/qr}-
3z
Importance yZUB8erb.
) &AZr(>
23 oizD:
|
The emphasis is on an implementation of a general approach to rule based decision "Z& {
making. PG&@.KY
Consideration / Attention T&w3IKb|}
Careful evaluation is necessary to ensure H2E!A2\m
Such a formulation does not change further considerations. &]16Hb~
Considerable attention has been paid to W*}q;ub;
Attention should be paid to an important finding of this investigation. $*9h\W-)`Q
Caution should be exercised in this process to avoid ... jF_K*:gQ
Primary consideration is given to ... components, though others can be accommodated )0yY|E
\
After ... has been defined by ..., a carefully analysis is carried out/performed to determine RUlM""@b
A number of factors such as ...need to be taken into consideration before making the =5[}&W
appropriate decision. kQsyvE
It should be noted that ~Q0jz/#c
It is important to point out that ... *&% kkbA
These considerations have heightened interest in the possibility of providing ... 9"I/jd0B
We should stress the fundamental importance of the xx vp[~%~1(
Results. kzbgy)PK3
Advantages / Disadvantage Pi40w+/
One of the major advantages of this new measure of xx is that it can be applied to the 4%KNHeaN
experimental study of Q/1
6D
One advantage of using a .. is the ease of preparing it. A+iQH1C0h
It has a very fast decision making process wHx1CXC
All the algorithms involve mostly logical operations. Q|"{<2"]U0
It can be easily and without additional cost implemented in a microprocessor;based DSM,dO'
environment. cr27q6_
It can reduce the waste of designing from scratch. =L
7scv%i
The advantages of using a xx to represent xx are the following: KNic$:i
However, xx is not without its shortcomings. : N> 5{
In most cases, the xxx shows an improvement over the existing xxx. TMhUo#`I|
Compared to the existing xx, the impacts of the xx are generally reduced by 5% to 9%. wbUpD(
The "best case" results shows a savings of 6% to 9%. Hd
U1gV>
Most of the existing works based on xx approach can only recognize a xx . `2o/W]SSk
Most of the above methods are computational expansive and limited to xx. bO>Mvf
Some other advantages of xx are the following: j;=+5PY
The problem is the limitation of this method to a limited domain of parts. 3G~ T_J&
It proved limited in application because it demanded precision in system modeling that _W@q
%L>
was impossible in practice. Y`uL4)hR5
There are advantages to be gained in the structuring of costs and benefits, the use of xx, vnw83a%3
The disadvantages of this method are also disadvantages of conventional xx approaches. zbOEF
This combines the best features of both techniques $-|`#|CBd
24 <*i
'
Hopefully, this tool can be as the reference framework of for developing a xx platform, ^.
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