Cryo-electron tomography (CET) is an imaging technique capable of visualizing the three-dimensional (3-D) structure of complex viruses, cells, and tissues in hydrated state. With the current resolution of 4-5 nm, CET can resolve supramolecular complexes that are responsible for many cellular functions. This paper discusses the important considerations in CET of biological specimens and identify areas where digital signal processing can make a decisive contribution. Topics discussed include the principles of electron tomography and CET, image background and feature contrasts in CET, acquisition and alignment of projection images, reconstruction of the image volume, denoising and segmentation of tomograms, and feature recognition in cellular tomograms

Curtin, P.T.

Gruenberg, D.

Hansen, K.S.

Hayes-Lattin, B.

Kovacsovics, T.J.

Lanier, K.S.

Leis, J.F.

Mauro, M.J.

Maziarz, R.T.

Menashe, J.I.

Segal, G.M.

Simic, A.

English

Leis, A.

Beck, M.

Gruska, M.

Best, C.

Hegerl, R.

Baumeister, W.

Leis, J.

MPG.PuRe

Cryo-electron tomography of biological specimens

Elsevier - Publisher Connector 

The failure of prognostic models (PM) for Hodgkins disease (HD) to predict outcomes after autologous stem cell transplantation (ASCT) 

J.F. Leis

K.S. Hansen

P.T. Curtin

B. Hayes-Lattin

K.S. Lanier

D. Gruenberg

M.J. Mauro

G.M. Segal

J.I. Menashe

T.J. Kovacsovics

A. Simic

R.T. Maziarz

Crossref

The failure of prognostic models (PM) for Hodgkins disease (HD) to predict outcomes after autologous stem cell transplantation (ASCT)

Leis, A. P.

Leis, J. W.

Max Planck Society eDoc Server

will improve survival in children with high-risk solid tumors. Fif-
teen patients were enrolled from March 2000 to August 2004 with
a median age of 6 years (2–19). Each had received at least 2 courses
of chemotherapy prior to study enrollment. Patients (Table 1) had
the following diagnoses: recurrent Wilms tumor (n  5), high-risk
Ewing’s sarcoma (n  3), recurrent hepatoblastoma (n  2), re-
current retinoblastoma (n  2), recurrent germ cell tumor (n  2),
and undifferentiated sarcoma (n  1). Patients were prepared with
etoposide (800 mg/m2/day on 6, 5, 4), carboplatin (667 mg/
m2/day on 6, 5, 4), and cyclophosphamide (1800 mg/m2/day
on 3, 2) prior to the ﬁrst transplant, and melphalan (60 mg/
m2/day on8,7,6) and cyclophosphamide (500 mg/m2/day on
5, 4, 3, 2) prior to the second. Children received a median
of 3.86  108 TNC/Kg (2.3–9.4) prior to the ﬁrst transplant, and
4.1  108 (2.5–10.5) TNC/Kg prior to the second. Eight patients
were in complete remission and 7 were in partial remission at the
time of ﬁrst transplant. One patient received local radiation instead
of the second transplant due to parental preference. Following each
rescue, patients achieved an ANC 	 500 at a median of 13 days
(11–34) and 12 days (10–31), respectively. Patients achieved a
platelet count 	20,000 at a median of 22 days (13–37) and 37 days
(15–113), respectively. There was a median of 38 days between
rescues (29–49). Five patients received radiation therapy post-
transplant to relapse sites. There were no toxic deaths. Toxicity was
primarily hematopoietic. Nine of 15 are surviving, 2 with recurrent
disease following tandem therapy at a median follow-up of 22 months
(11–55). Relapses following transplant occurred at the primary site
(n  3), distant (n  3), and primary/distant (n  2). Five have died
from progressive disease; 1 from late pulmonary toxicity. We con-
clude that tandem stem cell transplant used as consolidation in pa-
tients with solid tumors is feasible, well tolerated, and offers the
potential for cure in some patients with high-risk disease (Table1).
Table 1. Patient Characteristics
Patient Diagnosis
Stage/
Location
Relapse
Site Prior
to SCT
Relapse Site
Following
SCT
Follow-up
Post
Transplant
1 Undifferentiated
sarcoma
Pelvis NED,
55 months
2 Ewing’s Pelvis Primary
site, lungs
PD, died -
39 months
3 Hepatoblastoma Stage IV Primary site Alive, stable
disease -
53 months
4 Wilms Recurrent
stage IV
Primary site,
lungs
NED,
45 months
5 Ewing’s Pelvis Primary
site, lungs
PD, died -
11 months
6 Wilms Recurrent
stage I
Liver NED,
49 months
7 Wilms Recurrent
stage III
Lungs CNS Alive, stable
disease -
38 months
8 Ewing’s Metastatic NED,
41 months
9 Hepatoblastoma Recurrent
stage IV
Primary site Lungs PD, died -
22 months
10 Germ cell
tumor
Stage IV Pulmonary
toxicity,
died -
19 months
11 Wilms Recurrent
stage II
Primary site Primary
site
NED,
27 months
12 Wilms Recurrent
stage III
Primary site Primary
site
PD, died -
12 months
13 Retinoblastoma Bilateral Sinus, orbit NED,
16 months
14 Retinoblastoma Bilateral Pineal Pineal PD, died -
11 months
15 Germ cell
tumor
Pineal Primary site NED,
13 months
268
PHARMACOKINETICS (PK) OF 2 DOSING REGIMENS OF PALIFERMIN IN
PATIENTS (Pts) WITH HEMATOLOGIC MALIGNANCIES (HM) UNDER-
GOING HIGH-DOSE CHEMORADIOTHERAPY AND HEMATOPOIETIC
STEM CELL TRANSPLANTATION (HSCT)
Zia-Amirhosseini, P.1, Salﬁ, M.1, Elhardt, D.1, Aycock, J.1,
Cheah, T.C.1, Cesano, A.1, Hurd, D.D.2 1. Amgen Inc, Thousand Oaks,
CA; 2Wake Forest University School of Medicine, Winsto-Salem, NC.
For patients (pts) with HM undergoing HSCT, oral mucositis
(OM) is a frequent and debilitating complication that negatively
impacts treatment outcomes, patient quality of life, and healthcare
resources. Palifermin reduces the incidence and duration of severe
OM in the HSCT setting. This phase 1 open-label study assessed
the PK of 2 palifermin dosing regimens. Methods: Pts were 18 to
76 years old with HM and a Karnofsky performance score 70%.
Palifermin was administered intravenously once daily as follows: 60
mcg/kg/day for 3 consecutive days on day 11, day 10, and day
9 before conditioning (total body irradiation [TBI]  etopo-
side  cyclophosphamide) and following HSCT on days 0, 1, and
2 (part A) and a single dose of 180 mcg/kg (part B) before condi-
tioning on day 11 and after HSCT on d 0. In part A (6 total
doses), PK parameters were assessed after the ﬁrst, third, fourth,
and sixth doses. In part B (2 total doses), assessments were made
after each dose administration (day 11 and day 0). Results: In
part A, 13 pts received palifermin; in part B, 12 pts received the
single dose on day 11 and 11 pts received the single dose on day
0. For both dosing regimens, palifermin concentrations declined
rapidly (98% decrease) in the ﬁrst 30 minutes postdose, followed
by a slight increase in mean concentrations between 1 and 4 hours
and then a terminal decay phase. Respective mean (SD) PK pa-
rameter values for the 2 dosing regimens are shown in Table 1. In
part A, mean AUC0-t values were comparable between doses 1 and
3 (within 15%) and 1 and 4 (within 1%). In part B, mean PK
parameter values were similar (within 10% of each other) between
doses 1 and 2. The mean AUC after the ﬁrst 180 mcg/kg dose in
part B was approximately 4-fold higher than that after the ﬁrst 60
mcg/kg dose in part A. Mean half-life values ranged between 3.3 to
5.7 hours in part A and the value was 5.4 hours in part B. Con-
clusions:The PK data in pts receiving HSCT were consistent with
approximately dose-linear PK in the dose range of 60 and 180
mcg/kg, with no observed accumulation, based on AUC, after 3
daily doses of 60 mcg/kg in this pt population in the HSCT setting
(Table1).
Table 1.
Dosing
Regimen
(Dosing Day) n
AUC0-t
(hr  ng/mL)
Mean (SD)
Clearance
(mL/hr/kg)
Mean (SD)
Vss (mL/kg)
Mean (SD)
Part A - 60 mcg/kg/day  3 consecutive days
1st dose (day 11) 9 to 13 34.3 (15.9) 1730a (497) 5320a (2330)
3rd dose (day 9) 13 39.8 (36.4) - -
4th dose (day 0) 11 to 13 34.8 (22.5) 2030a (862) 3870a (2080)
6th dose (day 2) 13 21.2 (15.1) - -
Part B - 180 mcg/kg/day  1 day
1st dose (day 11) 12 140 (50.9) 1460 (600) 4290 (3270)
2nd dose (day 0) 11 143 (71.8) 1770 (1290) 4270 (4700)
Accurate computations of clearance (CL) and volume of distribu-
tion at steady state (Vss) were not possible for some concentra-
tion-time proﬁles.
269
THE FAILURE OF PROGNOSTIC MODELS (PM) FOR HODGKINS DISEASE
(HD) TO PREDICT OUTCOMES AFTER AUTOLOGOUS STEM CELL
TRANSPLANTATION (ASCT)
Leis, J.F.1, Hansen, K.S.2, Curtin, P.T.1, Hayes-Lattin, B.1,
Lanier, K.S.2, Gruenberg, D.2, Mauro, M.J.1, Segal, G.M.2,
Menashe, J.I.2, Kovacsovics, T.J.1, Simic, A.1, Maziarz, R.T.1 1.
OHSU Cancer Institute, Center for Hematologic Malignancies, Port-
land, OR; 2. Northwest Marrow Transplant Program, Portland, OR.
PM have been developed for patients with advanced HD in an
effort to identify high-risk individuals for and predict outcomes of
ASCT. Four PM were evaluated in patients transplanted in our
program between 1993 and 2005. One hundred and thirteen pa-
tients with relapsed or refractory HD received ASCT. Forty-ﬁve
patients received a conditioning regimen of busulfan, melphalan,
and thiotepa (BuMelTT), whereas 68 patients received other stan-
dard conditioning regimens (SCR) including Cy/TBI/VP (23),
CBV (39), and other (6). Followup is 113 weeks for BuMelTT
Poster Session II
95BB&MT
patients and 201 weeks for SCR patients. Factors differentiating
the BuMelTT and SCR groups included median followup post-
ASCT, total percent relapse after transplant, death after transplant,
relapse in prior radiation ﬁeld, and stage at salvage therapy pre-
ASCT. To date, 37% of the BuMelTT and 56% of SCR patients
have relapsed (P  .05) with a median time to relapse of 30 and 36
weeks (P  NS), respectively. Median OS, EFS, and RFS have not
yet been reached for BuMelTT patients compared with 439 (P 
.03), 67 (P  NS), and 74 (P  NS) weeks for the SCR patients
respectively. TRM was 5% in both groups at 5 years. Secondary
myelodysplasia has been seen in one patient transplanted with
BuMelTT and one with SCR. In multivariate analysis the
BuMelTT regimen and chemosensitive disease were associated
with improved OS (HR  0.19 and 0.31, respectively). Relapse in
prior radiation ﬁeld predicted for poor EFS and OS (HR  3.43).
We investigated the predictive value of 4 PM on the entire group,
BuMelTT and SCR cohorts. Each of these models has identiﬁed 3
poor prognostic indicators on the basis of multivariate analysis.
Patients with 0 or 1 factor were considered good risk and 2 or 3
factors poor risk. Only the SWOG model was predictive for OS for
the entire group (P  .01) and for the BuMelTT (P0.05) and
SCR (P  .01) cohorts. The SWOG model was also able to
differentiate EFS between good and poor risk patients for the
entire group (P  .02) and SCR group (P  .05) but not for the
BuMelTT cohort. None of the models could differentiate between
good and poor risk patients with regard to RFS. When good and
poor risk was deﬁned as 0 factors versus 1 to 3 factors, the RPCI
model was predictive of OS (P  .02) and EFS (P  .02) but only
for the BuMelTT cohort. The PM evaluated had limited utility in
our patient cohort with only the SWOG model being predictive of
outcome. Large multi-institutional studies are needed to improve
PM for transplantation in advanced HD.
270
PACLITAXEL FOLLOWED BY FILGRASTIM FOR PERIPHERAL BLOOD
STEM CELL (PBSC) MOBILIZATION IN PATIENTS WITH HEMATOLOGIC
MALIGNANCIES WHO EXPERIENCED PRIOR MOBILIZATION FAILURES
McKibbin, T.1,2, Burzynski, J.A.1,2, Twombly, R.G.1,2, Ochoa, J.L.1,2,
Tsai, T.W.1,2, Callander, N.S.1,2, Freytes, C.O.1,2 1. South Texas
Veterans Health Care System, San Antonio, TX; 2. University of Texas
Health Science Center, San Antonio, TX.
Cyclophosphamide followed by ﬁlgrastim (G-CSF) is frequently
utilized for PBSC mobilization in patients with hematologic ma-
lignancies, but is associated with adverse events such as hemor-
rhagic cystitis and a rate of febrile neutropenia as high as 30%. In
addition, white blood cell recovery varies among patients making it
difﬁcult to predict when to initiate apheresis. Our institution pre-
viously reported the efﬁcacy of paclitaxel followed by G-CSF for
PBSC mobilization in 30 patients with hematologic malignancies.
In 86% of those patients, paclitaxel mobilization was the initial
mobilization attempt. We report our experience with paclitaxel
250 mg/m2 IV once followed by G-CSF 10-16 mcg/kg/day in 27
consecutive patients treated at Audie L. Murphy Memorial Veter-
ans Hospital between 1/98 and 9/05. Twenty-one (78%) failed
mobilization with G-CSF 16 mcg/kg/day alone, 5 (18%) failed
mobilization with cyclophosphamide 3 g/m2 followed by G-CSF,
and 1 (4%) failed mobilization with rituximab, ifosfamide, carbo-
platin, and etoposide followed by G-CSF. Eight patients had mul-
tiple myeloma (MM), 17 had non-hodgkin’s lymphoma, and 2 had
acute myelogenous leukemia. Similar to our prior experience, 74%
of patients began apheresis on day 8 after paclitaxel administration,
based on a rising peripheral white blood cell count. The remaining
patients began apheresis on day 9 or 10 after paclitaxel adminis-
tration. Eighteen (67%) of 27 patients collected the target number
of CD34cells, deﬁned as 2  106 cells/kg. An additional 4
patients underwent PBSC transplant with fewer than 2  106
CD34 cells/kg (median 1.83, range 1.43&ndsh;1.98). One pa-
tient successfully collected adequate PBSC but elected to undergo
allogeneic PBSC transplant; thus 21 (78%) of 27 patients pro-
ceeded to high-dose chemotherapy followed by autologous PBSC
transplant. Median neutrophil (ANC 	 0.5  109/L) and platelet
(platelet20 109/L) recovery was 11 days (range 8–12) and 13 days
(range 9–24), respectively. Three patients (11%) developed febrile
neutropenia following paclitaxel resulting in one death prior to
high-dose chemotherapy in a patient with MM. Conclusions:
Paclitaxel followed by G-CSF is an effective mobilization strategy
for patients with hematologic malignancies who failed to mobilize
PBSC with other mobilization regimens. The low incidence of
febrile neutropenia and the predictable kinetics of white blood cell
recovery make this combination an attractive alternative for pa-
tients who fail to mobilize PBSC.
271
HIGH-DOSE CHEMOTHERAPY (HDCT) WITH HEMATOPOIETIC STEM
CELL TRANSPLANTATION (HSCT) IN PATIENTS WITH POOR PROGNO-
SIS EWING FAMILY TUMORS (EFT)
Bolotin, E.1, Shakhnovits, M.1, Cooper, L.1, Qian, D.1, Hitt, D.,
Sweetman, R.1, Sato, J.K.1, Miser, J.1, Forman, S.1, Rosenthal, J.1 City
of Hope National Center and Beckman Research Institute, Duarte, CA.
Objective: To evaluate the feasibility and safety of treatment
with two cycles of HDCT followed by autologous HSCT in
patients (pts) with poor prognosis EFT. A retrospective analysis
included 20 EFT pts treated at City of Hope National Medical
Center with 2 cycles of HDCT and HSCT from 1997 to 2003.
HDCT for the ﬁrst cycle consisted of melphalan (MEL)/carbopla-
tin (CARB) in 9 pts and Mel/busulfan (BU) in 11 pts. HDT for the
second cycle consisted of MEL/CARB (n  5); MEL/BU (n  3);
and cyclophosphamide/etoposide/CARB (n  5). A descriptive
analysis of the safety, rate of recurrence, transplant-related mor-
bidity (TRM) and mortality, event-free survival (EFS), and overall
survival (OS) are reported. Results: Out of 20 eligible patients, 13
(65%) received 2 cycles of HDCT followed by autologous HSCT.
Seven (35%) patients did not proceed to a second cycle as planned:
2 (28.5%) patients refused, 2 (28.5%) patients died of rapidly
progressive disease, 1 (14%) patient died of idiopathic pulmonary
ﬁbrosis, 1 (14%) patient had previous radiation, and 1 (14%)
patient had renal failure. The median follow-up (n  20) was 2.4
years (range 0.1–7.8 years). Grade 4 adverse events were limited to
mucositis and pancytopenia. No TRM was recorded after the ﬁrst
cycle of HDT and HSCT. There was one death after second cycle
due to rapidly progressive disease. A sustained absolute neutrophil
count (ANC 	 1000/mm3) for both cycles was achieved at the
median of 11 days. A sustained platelet count (	50,000/mm3) was
achieved at a median of 17 to 20 days. There were no differences
in OS rates between various HDCT regimens used and between
patients with or without metastatic disease disease at the time of
diagnosis. The Kaplan-Meier estimates of OS and EFS at three
years were 45% (CI 0.22, 0.69) and 47% (CI 0.25,0.70), respec-
tively. For patients who have received two cycles of HDCT fol-
lowed by HSCT, the Kaplan-Meier estimates of OS and EFS at 3
years were 58% (CI 0.30, 0.86) and 58% (CI 0.30, 0.86), respec-
tively. Conclusions: Dose-intensiﬁcation with two cycles of
HDCT and HSCT is feasible and relatively safe, with low and
acceptable treatment related morbidity and mortality. Adding a
second course of therapy does not impair engraftment. Both OS
and EFS compare favorably with previously published reports.
HDCT with HSCT may improve outcome in patients with poor
prognosis EFT. Further studies are required to establish the opti-
mal regimen for HDCT.
272
A LONG-TERM EXPERIENCE USING AUTOLOGOUS STEM CELL TRANS-
PLANTATION FOR THE MANAGMENT OF HODGKIN’S LYMPHOMA
Skikne, B.1, Divine1, C.1, Thudi, N.1, Deauna-Limayo, D.1,
Bodensteiner, D.1, Ganguly, S.1, Williams, C.2, Abhyankar, S.2,
McGuirk, J.2 1. The University of Kansas Medical Center, Kansas City,
KS; 2. Kansas City Cancer Center, Kansas City, MO.
Hodgkin’s disease (HD) patients generally have a good long-
term prognosis with approximately 70% cure rate. However, the
prognosis signiﬁcantly declines after disease relapse. High-dose
chemotherapy with autologous stem cell transplantation is an ef-
fective salvage option for patients in relapse with 5-year OS of
50%, and has become an accepted therapy. While transplantation
Poster Session II
96


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