18 =ZURh_{xV
Jurkat cells cultured in calcium-free RPMI-1640 medium (Gibco BRL; number {u
y^Bui}
22300-107) containing calcium-free 10% FBS were triggered by anti-Fas IgM. The g4,ldr"D
treated cells were harvested at the indicated time points and incubated with Fluo-3-AM at YL|)`m0-^5
a final concentration of 1 micromolar for 30 min at 37°C (Scoltock et al., 2000). The +zch e
labeled cells (50,000 cells per treatment) were then analyzed by exciting the cells at 488 Yn [
F:Z
nm and examining the fluorescence emission of Fluo 3 at 530 nm with a FACS Scan, /Q7q2Ne^*
(Becton Dickinson). A one micromolar concentration of LPA was used as a positive s/hWhaS<
control for Ca2+ induction. The data thus obtained was analyzed with the software Win , 2#Q>
MDI 2.8 and represented as contour plots.The effect of chelating intracellular calcium on zT)cg$8%fY
translocation of annexin I was studied by culturing Jurkat cells in the presence of 10 z0+JMZ/
micromolar BAPTA-AM, with or without the addition of anti-Fas IgM. Cells were r@r*|50
harvested and fractionated as detailed above, and the S-100 fractions were assessed by cOdRb=?9
immunoblotting for presence or absence of annexin I. y[l{
UBue:
Mouse bone marrow transduction and transplantation. dO2?&f
Retrovirus-mediated transduction of mouse bone marrow cells was done by published r3' DXP
methods49. Prestimulated low-density bone marrow cells were infected with high-titer `qmwAT
retrovirus supernatant on fibronectin-coated plates. Retrovirus supernatant was generated RjviHd#DXn
in the phoenix-gp cells with a mouse stem cell virus–based retroviral vector coexpressing iT{4-j7|P4
EGFP and HA–RhoH as described50. EGFP+ sorted cells were transplanted by g)Hsd0
intravenous injection into the sublethally irradiated (300 rads with a 137Cs irradiator) /y{fD
CC
Rag2-/- recipient mice. At 9 weeks after transplantation, thymus, peripheral blood, bone p2l@6\m\
marrow, spleen and lymph nodes from each recipient mouse were collected for analysis R&t2
of EGFP+ chimerism and hematopoietic lineage by flow cytometry. Expression of Y{e,I-"{
HA–RhoH and HA–RhoHF73F83 in EGFP+ sorted thymocytes of recipient mice was Oz\J
+
confirmed by immunoblot analysis. Ag:/iB]
Determination of renal morphology ]6v7iuvI
Kidney slices were postfixed in buffered 2% OsO4, dehydrated, and embedded in an @(=?x:j
Araldite-EM bed 812 mixture. Large sections were cut perpendicular to the renal capsule, }T}c%p
containing cortex, and medulla. Thin (1 m) sections were analyzed in a blinded manner xG
edY*[`
for morphologic alterations, as previously detailed Tg@G-6u0c
Patient population g8Y)90 G
Patients included in the study met the following criteria: (1) biopsy-proven IMN; (2) `xv Uq\
creatinine clearance 30 ml per min per 1.73 m2; and (3) persistent proteinuria >5 g per +7WpJ;C4
24 h despite treatment with an HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A) U<XfO'XJ
reductase inhibitor, an ACEi, and/or ARB at maximal tolerated dose for at least 4 months. *j,noHUT~>
The Mayo Institutional Review Board and the Research Ethical Board, University Health (]mBAQ#hw
Network, University of Toronto approved the study protocol. All patients gave written M^l%*QF[,q
informed consent. Patients who had been on treatment with prednisone, cyclosporine, or t`}=~/#`X
19 ?t\GHQ$$?
mycophenolic mofetil within the last 4 months or alkylating agents within the last 6 2XBHo (
months were not included in the study. Patients with active infection, diabetes, or a E<B/5g!
secondary cause of MN (for example, hepatitis B, systemic lupus erythematosus (SLE), #+"D?
medications, malignancies) were also excluded. IYptNR
Treatment <uoVGV5N
At enrollment, a low-sodium (<4 g day-1) and low-protein (0.8 g per kg per day of AxJqLSfyb,
high-quality protein) diet was recommended and patients were encouraged to maintain OS
L~a_
the same diet throughout the duration of the study. All patients received a similar 0V"(}!=2a
conservative treatment regimen that included loop diuretics to control edema, an 60|m3|
0o
HMG-CoA reductase inhibitor, and an ACEi combined with an ARB if tolerated. a&mL Dh/
-Blockers and non-dihydropyridine calcium channel blockers, in that order, were added t]3:vp5N]
when required to control systolic blood pressures to <135 mm Hg in >75% of the 9ygNJX'~
readings. Patients who after a minimum of 4 months of conservative therapy and %dmQmO,
maximized Ang II blockade had proteinuria >5 g per 24 h received two i.v. infusions of Bz,Xg-k+
rituximab at a dose of 1000 mg on days 1 and 15. To minimize infusion reactions, )WEOqaR]
patients were premedicated with acetaminophen (1000 mg) and diphenhydramine vXdI)Sx[
hydrochloride (50 mg) orally. In addition, methylprednisolone (100 mg, i.v.) was given DvI^3 iG8
prior to the first rituximab infusion. B-cell depletion was defined as CD19+ count - t4F
<5 cells per l at any time and B-cell recovery was defined as CD19+ cell count ] : ](xW%
>15 cells per l. Patients treated with rituximab, who at month 6 had proteinuria >3 g per tC+11M
24 h and in whom CD19+ B-cell counts had increased to >15 cells per l, received a 7|ACJv6%9
second course of rituximab treatment following the same protocol described above. FhEfW7]0,
Follow-up 'OhGSs|
In all patients, clinical and laboratory parameters including complete blood counts, Oeg^%Y
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum 7
0R_O&f-k
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry uXGAcUx(
and at months 1, 3, 6, 9, and 12. Creatinine clearance and protein and creatinine excretion tOQnxKzu
in the urine were assessed by performing two consecutive 24-h urine collections at each >#;>6q9_
time point. Data were considered accurate when urinary creatinine excretion was oBPm^ob4
consistent with a complete 24 h collection. The mean of the two measurements was y]k{u\2A
considered for the analysis. The presence of HACAs was evaluated at baseline and at (3m^@2i
months 3, 6, 9, and 12. tg2+Z\0)4g
Method / Approach / Study/ Technique 5cr\ JR
A discussion is presented of a problem-solving system eN TKX
To improve the efficiency of the method, the following approach may be applied. N71%l
In order to an investigation was made to find the causes of the 7X/B9Hee
Although large collections of rules and equations have been complied, none are generally ~NK|q5(I
accepted z_$F)*PL
20 U]E~7C
This approach will be explained and discussed thoroughly in the body of the report. ?j8_j
This can be accomplished by R36A_
This algorithm to compute the total cost can be described step by step as follows: FCt %of#
The above preliminary analysis has provided important information ,V+,3TT
Various methods have been proposed for selecting an optimum... ~N+H7T.L
These concepts have been applied to s";9G^:
On the basis of the concept mentioned above, N*}g+IS
This can be achieved by g % 8@pjk
In addition, tissues were stained for infiltrating lymphocytes (CD20 and CD3), and the HhqNpU
amount of interstitial fibrosis was quantified by histomorphometry. Formalin-fixed, Ij7[2V]
c
paraffin-embedded sections were cut onto coated glass slides. Following heat-induced mv`ND&
antigen retrieval, sections were incubated at 20°C overnight with either anti-CD20 JHsxaX;c
primary antibody or anti-CD3 primary antibody, both at 1:1000 dilution (Dako, Canada 6PRP&|.#
Inc, Mississauga, Ontario, Canada). After rinsing all sections, pretreatment with 3% y?xFF9W@H
hydrogen peroxide was performed to prevent endogenous peroxidase activation. Sections 1n>(CwLG"
were incubated with a secondary rabbit anti-mouse antibody linked with avidin–biotin mA\}zLw+r9
complex. Sections were counterstained with hematoxylin and examined by light `mDCX
microscopy. 2?(dS
The HACA assay is a proprietary bridging enzyme-linked immunosorbent assay &jE@i#
performed at Genentech Inc. that measures the antibody response to rituximab in human 67P@YL
serum samples. KyRcZ"
In all patients, clinical and laboratory parameters including complete blood counts, $xq$
electrolytes, serum albumin, B-cell flow cytometry for CD19+ B cells, serum \2y[Hy?
immunoglobulin (IgG, IgM, IgA) levels, and a lipid panel were evaluated at study entry ,-5|qko=
and at months 1, 3, 6, 9, and 12. @A8@j%CK1
This fact suggests that a new concept o#=O5@>ai
This was accomplished by taking ... 2$yNryd
The preparatory stage is very time consuming process. 1C<@QrT
Test are performed for validity, completeness, and compatibility ECvTmU'=
There is little hope of achieving successful ... /yTPb
There has been an increasing awareness of the potential of using most ..so far made have eDS,}Z'
not taken this approach, with the exception of )cBO_
Only a few studies can be found. cPPTGpqw
It is a very tedious process to go through A $W~R
It is only when .. has been completed that .. may be effected CjUYwAy$k
The entire interpretation process is conducted in one's head. C?t!Uvs
These approaches are sometimes very tedious. c4(og|ifk
Several techniques can be used #9p|aS\
A polynomial parametric model can be written as [the following]/[follows]: ! +XreCw
A xx model is constructed/formulated using xx. \JIyJ8FleC
A xx model represents an xx by its xx. IQQ QB
A process decision model captures the logic essential to @ x5LrQ_`r
From the equation above, xx is equal to the summation of xx times the ... E{{Kzr2$
21 =&roL7ps
The validity of a xx model can be checked using Euler's formula. :HkXsZ
Given a model, one can mathematically determine whether ... or ... "WzD+<oL
Equations for xx need to be derived and implemented in the system. Nyl)B7/w
A number of heuristic rules have been developed for CX#d9
8\b
Optimum .. techniques can be made more reliable by ... so that !:M+7kmr7t
An algorithm based on the characteristic ... is used to determine 9i+`,r
Euler's formula states the following: QfRo`l/V9
The completed model should agree with the formula. !AN;
For manufacturing purposes, a detailed and precise model of the object is necessary U}DLzn|w
Engineering design models are very well defined; therefore, cr ~.],$Om
To keep the domain narrow enough to be implementable, yet wide enough to be useful. qpV"ii
Point of View nfksi``Vq
from an implementation standpoint, 2&XNT-Qm
From the point of view of this application, Ih)4.lLcKn
From this point of view, Zadeh suggested an inference rule named xxx (CRI for short). z"f+;1
Information is the meaningful interpretation and correlation of some aggregation of data
%BF,;(P
in order to allow one to make decisions. [G'
+s
From a practical point of view, the computational aspects of an FLC require a
]EqwDw4
simplification of the fuzzy control algorithm. y+nX(@~f]
The use of a hammer to insert screws, although partly effective, tends to distort, destroy, 2=uwGIF
and generally defeat the purpose of using a screw [Kusiak AI Implications for CIM kO4'|<
p.129] _fS4a134R
Statistical analysis Sm;@MI<@/
Data were analyzed by one-way analysis of variance comparing the three conditions *U +<Hv`C
(sham operation, ischemic AKI, and bilateral nephrectomy) at each time point. If Ako]34Rl,
significant F-statistic from analysis of variance existed, this test was followed by Dunnett ch33+~Nn
post hoc multiple comparison procedure with sham operation as the control group. For all JT6}m
other comparisons, Student's t-test was used. A P-value of <0.05 was considered as r+;k(HMY}[
statistically significant. Ko0?c.l
Values are expressed as means s.e.m. and significance was evaluated by Mann–Whitney 7[I +1
U test using GraphPad Prism, version 4.0 software (GraphPad Software Inc., San Diego, jXPbj.
CA, USA). #HZ W57"
All values are expressed as means s.d. Statistical significance (defined as P<0.05) was oxL)Jx\c9A
evaluated using analysis of variance and Bonferroni t-tests, and the two-tailed Pearson's }46Zfg\T6n
test, where appropriate. {.ypZ8JU
Data are expressed as mean s.d., median and interquartile range, or frequencies, as 7^tYtMm|U
appropriate. Variables who deviated from the normal distribution (positively skewed) KW$.Yy
were log-transformed (log10) before the correlation study. 2#/23(Wc
Data are represented as means s.e.m. Student's t-test and multiple comparisons with t-test EvH/d4V;
post hoc analysis of variance were used as indicated below, for the comparison of vo(:g6$
22 gegM&Xo
morphological, immunohistological and functional parameters. Statistical significance uh`5:V
was set at P<0.05. TbLU[(m-n
The primary efficacy parameter was defined as change in urinary protein excretion from (4?^X
baseline (week 0) to 12 months after treatment. The 12-month changes were tested e+)y6Q=
against zero using the paired t-test. Secondary end points included 6-month changes in g#`}HuPoE
protein; the number with PR or CR at 6 or 12 months; and changes in glomerular @Ne&%F?^Z
filtration rate (GFR), serum albumin, and lipid profiles. Study sample size was based on Kj)sL0
the desire to have 80% power to detect a drop in urinary protein of at least 2.0 g day-1. a,(nf1@5
Assuming a two-sided hypothesis test performed at a significance level of 0.05 and an s.d. (V1;`sI8
of urinary protein change of 2.5 g, it was determined that 15 patients were required.2 :{4C2qK>
Definition of remission status is according to the criteria established by Cattran et al.30 [ r
CR was defined as proteinuria <0.3 g per 24 h, PR as proteinuria 3 g per 24 h, and a }Xfg~%
6
>50% reduction in peak proteinuria and non-response as <50% reduction in peak ]=9 d'WL
proteinuria. Any patient reaching a CR or PR was considered a treatment success N;,zPW a
The statistical significance of differences for the mean values of cytokine concentration UBJYs{zz
and T cell proliferation was determined with Student's t-test. Differences with a P value h(R7y@mp\0
of less than 0.05 were considered significant. Zb2PFwcy
Statistical significance was determined by Student's t-test. Data with a P value of less dm]g:KWg
than 0.05 were considered significant. |?ZU8I^vW
In vitro and in vivo experiments were analyzed by the two-tailed Student's t-test, with P Ijap%l1I
values less than 0.05 considered statistically significant. -2!S>P Zs
The significance of differences among groups was determined by Student's t-test and `Tf}h8*
one-way analysis of variance (SigmaStat software; Jandel). M$Bb,s
Comparisons egBk7@Ko
As well, the pros and cons of these representations from a process planning point of view ?@Q0;LG
will be discussed. $${3I4
The method of using xx to implement xx described by Zadeh (1973) appeared more h Ypj
suitable L)&^Pu
As discussed [in the previous section]/[preciously], otfmM]f
Relation po\jhfn
We can not invert F' directly because it defines a many-to-one mapping. U<;{_!]
The relationships appear very complicate S7WHOr9XMV
Lifting tasks involve complex and imprecise relationship between the task variables and ]n>9(Mp!M
the human operator's characteristics. ]kx<aQ^
These methods are based on the relationship between ... and ... -s!J3DB
The fundamental concept of a fuzzy rating language is that we can establish a relationship l)o!&]2
among terms such as high, medium, and low, and then modify these relationships. Z.!<YfA)
This article will thus mention the latter as well as the former. qo7<g*kf~
The former two bear a close relation to a fuzzy Cartesian product. 5'+g[eNyBV
Importance R.2i%cU
) .a :7|L#a
23 74:( -vS
The emphasis is on an implementation of a general approach to rule based decision gY[G>D=
making. Jk`Jv;
Consideration / Attention wJJ|]^0.
Careful evaluation is necessary to ensure _<jU! R
Such a formulation does not change further considerations. sC[#R.eq
Considerable attention has been paid to ?lgE9I]
Attention should be paid to an important finding of this investigation. CQ<