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									                                                            *Clinical Pathology Dpt., Faculty of Medicine, Cairo
Chapter 9                                                   University.
                                                            Egypt J Lab Med 12: 131-143, 2000 (Proceedings of
                                                            11th Conference)
                                                            This study included 44 cases [21 cases AL in
                                                            hematological remission i.e. 0-1% blast cells in B.M.
                                                            smear (11 ALL and 10 ANLL) and 23 NHL presumed
9.1. Decreased incidence of Acute and Chronic               not disseminated (either in P.B. or B.M.)]. The tests
Graft Vs. Host Disease Following Primary                    done were direct blood and B.M. smears, buffy coat of
Transplantation of Allogeneic Peripheral                    blood and B.M., bone marrow culture as well as B.M.
Blood Stem Cells in Patients With Severe                    trephine biopsy for the NHL group. In AL cases, 13
Aplastic Anemia                                             showed relapse on follow- up (5 ALL and 8 ANLL).
O.A. Fahmy, H.K. Mahmoud, A. El Haddad*, A.                 This could be detected by P.B. buffy coat scanning in
Kamel*, M. Nazih*, A. Sobhy*, T.S. Rabie*, M.A.             1/13 (7.6%) and by B.M. buffy coat scanning in 13/13
Hassaballa*.                                                (100.0%). Such cases had 2% or more blasts. Bone
BMT Team, Nasser Institute, Cairo, Egypt.                   marrow culture showed blast colonies in 1/5 (20.0%) of
Blood 96(11): 400a, 2000                                    the relapsed ALL cases and in 8/8 (100.0%) of the
                                                            relapsed ANLL cases. In Non Hodgkin's
Graft vs. Host Disease (GVHD) is still one of the           Lymphoblastic Lymphoma presumed not disseminated
leading causes of morbidity and mortality following         in P.B. or B.M., 11/23 showed dissemination on
allogeneic hematopoietic stem cell transplantation. A       follow-up. This could be detected by P.B. buffy coat
concern is still going on regarding the incidence of        scanning in 9/11 (81.8%) of the disseminated cases, by
chronic GVHD in patients allografted with peripheral        B.M. buffy coat scanning in 10/11 (90.9%) and by B.
blood stem cells (PBSCs). We have already reported          M. trephine biopsy in 11/11 (100.0%). Of the 23 Non
the feasibility of allogeneic PBSCT in 18 heavily           Hodgkin's Lymphoblastic Lymphoma cases presumed
pre-transfused patients with severe aplastic anemia         not disseminated, 10 were also subjected to B.M.
(SAA), and showed that the incidence of acute GVHD          culture. Eight of 10 showed dissemination on
is not increased, if not even less, as compared to          follow-up. Bone marrow culture showed colonies in
patients receiving marrow cell. In the current study, we    2/8 (25.0%) of the disseminated cases. From this study,
are reporting on the incidence of chronic GVHD in the       it is concluded that there is a gradual increase in
same cohort of patients. In the period between July         efficiency for the detection of invasion or relapse front
1997 and November 1998, 11 males and 7 females              scan of P.B. buffy coat to B.M. buffy coat to B.M.
(mean age, 19) received Filgrastim mobilized PBSCs          culture in both AL and NHL and trephine biopsy in
from their HLA-identical siblings, and all were             NHL. Trephine biopsy was found to beg superior to
conditioned with Cyclophosphamide + ATG. The mean           B.M. culture in the NHL group. These cheap tests are
number of infused unmanipulated CD34+ve cells was           proposed to be done for all AL and NHL cases as their
7.5x106/kg body weight. All patients received GVHD          routine follow-up. These cover almost all early
prophylaxis using Methotrexate + Cyclosporine. The          relapsing; or early invaded cases especially if
tempo of hematopoietic reconstitution was rapid (mean       molecular biologic detection is not available.
number of days for neutrophil and platelet recovery
was 12 and 15, respectively), and no single early or late   9.3. A New Chemical Technique For
graft failure was reported. The main causes of death        Quantitative Estimation of Bone Marrow Iron
included intracranial hemorrhage (n=1), CMV                 Concentration (BMIC) As Compared With The
infection (n=l), interstitial pneumonitis (n=1), sepsis     Cytochemical Staining For Iron Storage.
(n=I), late onset infectious meningitis (n=1) and acute     Application      in     Different    Hematologic
GVHD (n=2). No other cases of acute GVHD was                Malignancies
observed. During a follow-up period of up to 36             Mostafa I. Aboul Enein, Nahed Abd El. Wahab,
months, only one patient (5.5%) developed chronic           Badawy M. Al Kholy*, Khaled M. Aboul-Enein, Ahmed
GVHD, which was limited and well controlled under           S. Bedeir and Khaled A. Emara.
immunosuppressive therapy. In conclusion, our data          Egypt J Lab Med 12(2): 413-421, 2000
suggest that in patient with SAA, allogeneic PBSCT is
not only associated with rapid engraftment but also         This study presents a new chemical method for
with decreased incidence of both acute and chronic          estimation of BM iron concentration (BMIC). It is
GVHD.                                                       comparable to the cytochemical Prussian blue method,
9.2. The Comparative Sensitivity of The                     but is easier, simpler and more accurate. The value of
Different Techniques Used For The Detection                 this chemical method is apparent when applied to
of Early Relapse of Acute Leukemia and                      normal and malignant cases. Decreased, normal and
invasion of Lymphoblastic Lymphoma                          increased B.M. iron was clearly verified. It is proposed
Mostafa I. Aboul-Enein, Khaled M. Aboul-Enein,              to generalize the use of this method for accurate and
Nahed Abd-Elwahab, Maha Saleh, Sonya F. Arsanios            clear BM iron storage evaluation.
and Ahmed A Shams El-Din.
Clinical Pathology Dpt., NCI, Cairo University,             9.4. Acquired Von Willebrand Disease in
Scientific Annual Report 2000-2002

Hematologic Malignancies At The National                     Sonya F. Arsanyos; **Ola M. Eldessouki*
Cancer institute (NCI) Egypt                                 *Clinical Pathology Department, National Cancer
Magda M. Assem, M.D.*; Mohamad A.M. Aly,                     Institute, Cairo University, **Clinical Pathology
Ph.D.**; Ibrahim Abdel Salam, Ph.D.**; Mohamad               Department, Cairo University Hospital
Hassan Shaheen, M.D.****; M. Nagib Zuhair,                   Egypt. J. Lab. Med. 12 (3): 635-646, 2000.
M.D.****; Maha Saleh, M.D.*; Nasr M. Aly,
M.D.****; Samia Y. Akle, M.D.* and Walid Radwan,             This study involved (60) cases: (15) ALL cases (7
M.D.                                                         active disease and 8 in remission), (17) ANLL cases
Departments of Clinical Patltology*, Tumor Biology**         (10 active and 7 in remission), (20) CML cases (7
& Medical Oncology****, NCI & Clinical Pathology,            treated and 13 untreated), (8) treated CLL cases and 10
Kasr El-Aini ****, Cairo University, and Hematology          normal individuals (as a control group). In all cases,
Department, Military Medical Academy. ***                    serum sL-selectin level was measured. The CSF level
J Egypt Nat. Cancer lnst 12(2): 117-723, 2000                was assessed in ALL cases. In ANLL cases mean
                                                             serum sL-selectin level in disease was (12.23±8.37
Acquired von Willebrand disease (AvWD) is an ac-             ug/ml) which was not different from those in remission
quired bleeding disorder, which may suddenly become          (mean 12.9±11.52 ug/ml). Both were higher than
manifest in individuals, in the absence of a past            normal control (mean 117±0.73 ug/ml). In ALL cases:
personal or family history of bleeding and frequently in     patients in active disease showed a higher serum sL-
association     with     monoclonal        gammopathies,     selectin level (mean 20.6±10.48 ug/ml) than those in
lymphoproliferative,        myeloproliferative         and   remission (mean 10.13±5.58 ug/ml) but this difference
autoimmune disorders. In a minority of cases AvWD            was statistically insignificant (p=0.074). Both were
may develop in association with drugs or solid tumors.       much higher than the normal control (mean 1.7±0.73
One hundred and fourteen patients attending to the           ug/ml) .There was also a strong correlation between
Medical Oncology Department, NCI, presenting with a          serum sL-selectin level and percentage of peripheral
recent development of non-thrombocytopenic mu-               blood blast cells (p<0.01). The serum selectin level was
cocutaneous bleeding were included in the study. Due         significantly higher than the CSF level (p=0.001) and
to the limitations and variability of each assay and         the CSF level was not correlated with CNS
because of AvWD heterogeneity, no single test                involvement in CML cases: the serum sL-selectin level
procedure was sufficiently robust to permit detection of     was significantly increased in untreated cases (mean
all subtypes. However, this is not the case in AvWD,         27.73±12.39 ug/ml) than treated cases (mean
since the most common pattern observed is deficiency         12.47±10.11 ug/ml) (p= 0.034). In CLL cases: All
of large vWF multimers 20/39. In this regard, the            cases were treated but with a mean value of serum sL-
multimer analysis would depict most of the cases. Four       selectin of 49±23.41 ug/ml, which is significantly
separate assays for vWF were included: vWF:                  higher than that of normal control (p<0.05). The mean
Multimer analysis, vWF; Antigen (vWF: Ag), factor            value of serum sL-selectin in active CML cases
eight coagulant activity (FVIIIc) and Ristocetin             (27.73±12.39 ug/ml) was significantly higher than its
cofactor (vWF: RiCof). 39/114 cases were diagnosed           level in active acute leukemias (15.68±9.93 ug/ml)
as AvWD (17 CML, 11 NHL, 8 CLL & 3 MM). The                  (p=0.003). We therefore propose that monitoring of
39 cases of AvWD were subclassified: 3 cases as type         serum sL-selectin level is more sensitive to determine
1, 20 cases as type 2A, 8 cases as probable type 2N, 7       disease state in chronic leukemias and ALL cases than
cases as prohahle type 2M (based on the low Ri:              AML cases. Its va1ue in serum or CSF in ALL patients
Cof/vWF: Ag ratio <0.36) and 1 case as AvWD                  needs further study with larger number of cases.
(unrecognized subtype). The increased incidence of
AvWD in CML patients was statistically significant           9.6. A Simple Strategy For Breakpoint
(p=0.04). Using receiver operator characteristic (ROC)       Fragment Determination in Chronic Myeloid
analysis, the best diagnostic single test wits found to he   Leukemia
RiCof, followed by FVIIIc, APTT and lastly vWF: Ag.          Azza M Kamela, Heba M Shakera, Farida H.
Although patients with blood group O were most               Gadallaha, M Reda Hamzab, Othman Mansourb, Omar
frequently and significantly (p=0.046) affected when         H El Hattabc, Heba S. Moussaa
the multimer test was used, this turned insignificant          Department of Clinical Pathology, National Cancer
when the Ri:Cof test was analyzed. This could be             Institute, Cairo University, Cairo, Egypt; b Department
relevant to the normal distribution of blood groups          of Medical Oncology, National Cancer Institute, Cairo
among the population rather than to the inherent low         University, Cairo, Egypt; c Biostatistics and
level of vWF:Ag in blood group O individuals. This           Epidemiology Unit, National Cancer Institute, Cairo
report emphasizes that AvWD is not an extremely, rare        University, Cairo, Egypt
disorder, particularly in the setting of hematologic         Cancer Genetics and Cytogenetics 122: 110-115, 2000
malignancies in Egypt.
                                                             Molecular characterization is considered a part of the
9.5. Comparative Analysis of Serum SL-                       routine work-up of chronic myeloid leukemia (CML)
Selectin Level in Acute and Chronic                          cases. Southern blot analysis using the universal BCR
Leukaemias, Relation To Disease Activity and                 (UBCR) probe on Bg/II-digested DNA samples is the
Correlation With CSF Selectin Level in ALL                   most commonly used technique, while employing the
*Moustafa I. Aboul Enein,* Lobna M. Abdel Megid,*            human 3’ bcr probe (PR-1) is usually considered a

                                                                                                    9. Leukemia

complementary tool. In this study, we tried to develop    normal in all disease states in both AML and ALL. The
a simple and economic strategy for molecular              mean ferritin level was high in new cases, remission
characterization of CML using the 3’ probe as it has      and relapsed patients whether in AML or ALL as
been shown to be the one capable of locating the          compared to normal control. The least increase in both
breakpoint site. Seventy-eight cases of CML were          AML and ALL was in relapsed patients, yet it did not
studied. Molecular analysis was performed using the       reach the level of significance. In conclusion, serum
Southern blat technique. DNA was digested with            ferritin has no diagnostic or prognostic value in acute
BamHI, Bg/II, EcorI, and XbaI. Hybridization was          leukemia due to its low specificity and sensitivity.
performed using the human 3’ bcr (PR-1) probe.            Bone marrow iron in both AML and ALL cases: serum
BamHI and Bg/II could differentiate fragment 1 (Fl)       iron did not correlate with increased bone marrow iron
showing rearrangement (R) with BamHI and germline         storage. The iron reutilization defect represents
configuration (G) with Bg/II; F2/3 showing R with         quantitatively the most important basis for the anemia
both, and F4 showing R with BamHI and G with Bg/II.       of malignancy. CSF ferritin can be used as a marker of
P2/3 cases were further divided by HindIII enzyme into    CNS invasion, as the mean CSF ferritin in cases with
P2 showing (G) and F3 showing (R). Fragment 0             blastic invasion was almost double that of cases not
showed G with both, but R with EcorI and/or XbaI,         invaded. CSF ferritin post CNS therapy in previously
while 3’ deletion gave G with all four enzymes. Our       invaded cases was markedly high.
results showed a relative incidence of 6.4% for FO,
20.5% for Fl, 32.1% for P2, 19.2% for F3, 15.4% for       9.8. Successful Primary Transplantation of
F4, and 6.4% for 3’ deletion. Sixty cases were            Allogeneic Peripheral Blood Stem Cells in
evaluated clinically and hematologically and were         Patients With Severe Aplastic Anemia
followed up for disease evolution and survival. They      Fahmy, O.A.; Mahmoud, H.K., Haddad, A.E.; Kamel,
included 32 cases in early chronic phase, 24 in late      A.M.; Nazih, M.A.; Sobhy, A.M.; Rabie, T.S.
chronic phase, two in acceleration, and two in blastic    Nasser Institute, Cairo, Egypt.
crises. No significant correlation was encountered        ASBMT, March 29-April 1st, 2000, California, USA
between the breakpoint site and any of the clinical and
hematological data except those patients with 3’          Allogeneic peripheral blood stem cell transplantation
deletion who showed a very short survival. The study      (PBSCT) is gaining more popularity in the treatment of
emphasizes Southern blotting as the method of choice      different hematologic disorders. However, apart from a
for molecular characterization of CML and offers a        few cases, no data exist regarding the feasibility of
simple and economic strategy for diagnosis and            primary allogeneic PBSCT in heavily pre-transfused
determination of breakpoint fragment.                     patients with severe aplastic anemia (SAA). Between
                                                          July 1997 and November 1998, we have transplanted
9.7. Changes in Iron Metabolism and Types of              18 patients with SAA using allogeneic PBSCs. They
Anaemias During The Course of Acute                       were 11 males and 7 females (mean age 19). All
Leukaemias and The Possible Use of CSF                    patients received Cyclophosphamide + Antithymocyte
Ferritin As A Marker or CNS invasion in                   globulin (ATG) as the conditioning regimen. The mean
Egyptian Cases                                            number of Filgrastim - mobilized CD34+ cells from the
Mostafa I. Aboul-Enein, Amira M. Khourshid, Farida        HLA-identical siblings was 7.5 X 106/kg recipient
H. Gadallah, Khalid Aboul-Enein, Azza H. El Sissy,        weight. All patients received Methotrexate +
Ashraf Khyri Zaki* and Ahmed S. Bedeir                    Cyclosporine for graft versus - host disease (GVHD)
Department of Clinical Pathology, NCI, Cairo              prophylaxis. The mean number of days for neutrophil
University,* Department of Medical Oncology, NCI,         and platelet recovery was 12 and 15, respectively. Six
Cairo University                                          patients (33.3%) died; the main causes of death were
Egypt J Hematol 25(3): 349, 2000                          intracranial hemorrhage, interstitial pneumonitis, CMV
                                                          infection, acute GVHD and sepsis. The remaining 12
This work involved 82 cases of acute leukemia             patients are alive and well for up to two years after
(43AML & 39 ALL) and 13 normal controls to study          transplantation, with no single case of early or late
iron metabolism. 40 cases and 10 normal controls were     graft failure. One of these is having a well-controlled
evaluated for CSF ferritin. In AML cases: The mean        limited chronic GVHD. Thus, our data suggest that
iron level was high (205.6±122.8 mg/dl) at                allogeneic PBSCT is quite feasible in heavily
presentation. During remission: The mean iron level       pre-transfused patients with SAA and provides a faster
was lower (154.1±97.6 mg/dl) compared to that at          reconstitution without increased incidence of acute
presentation (yet it was within the normal range).        GVHD compared to bone marrow transplantation
During relapse: The mean iron level was even lower        (BMT). Further follow up is needed to determine die
(88.5±69.54 mg/dl) compared to that at presentation.      ultimate incidence of chronic GVHD.
This may be of prognostic value, but did not reach the
level of significance. In ALL cases: there was no         9.9. Recommended Procedures For The
changes in the mean iron level at presentation            Classification of Acute Leukemias
(l83.2±134.4      mg/dl)    and     during    remission   O. El-Diwanyl and M. Assem2,
(l85.43±139.2 mg/dl), but during relapse, the mean iron    Al Azhar University, Cairo, Egypt and 2National
level was lower (125.07±124.37 mg/dl) compared to         Cancer Institute, Cairo University, Cairo, Egypt
that at presentation and remission. The mean TIBC was     The 28th World Congress of ISH, August 26-30, 2000.

Scientific Annual Report 2000-2002

Toronto, Canada. Abstract #17815                            of bcl-2 and bax expression in CLL patients was sig-
                                                            nificantly higher than in ALL patients p=0.003 and
The classification of acute leukemias is widely based       0.030, respectively. The level of bcl-2 expression in
on a combined morphological, cytochemical and               CLL patients was significantly higher than in ALL
immunophenotyping approach. The International               patients and control group, p=0.006. Our results
Council for Standardization in Haematology (ICSH)           confirmed a higher sensitivity of WB in detection of
converted an expert panel to consider currently             these cell cycle proteins when compared to IC. Higher
available diagnostic techniques with the aim of             bcl-2 expression was more evident among ALL
defining a minimum ctyochemical and immunological           patients above 18 years, p=0.03. The level of bax
diagnostic panel (intracellular phenotyping and             expression was significantly higher in patients with
membrane immunophenotyping) that could be used as           total leukocytic counts (TLC) ≥50 x 109/L, p=0.02.
core components for the classification of acute             Although no relation was found between bcl-2 and bax
leukaemias such as myeloperoxidase (MPO),                   expression and an increased probability of relapse yet,
chloroacetate esterase (Chlore E) and α-naphthyl            overexpression of bax (but not bcl-2) was associated
acetate esteralc (ANAE). Subsequent immunological           with an unfavorable disease outcome, p=0.02. In cases
analyses for cytoplasmic CD3, CD22, and TdT can be          with strong expression of bcl-2, an over expression of
done for cases of acute leukemias that remain               bax as well was detected by WB. This finding reflects
unclassified by primary cytochemistry. Cytogenetic          the fact that it is the balance rather than the level of
and DNA genotyping were excluded from the panel             either protein product that determines the influence of
used because of their restricted availability. The use of   these cell cycle proteins on prognosis of ALL.
these minimum primary cytochemical and intracellular
phenotyping procedures will lead to consistent              9.11. The Expression of P53 Protein in Chronic
classification of most acute leukemias and finally          Lymphocytic Leukemia Is Associated With
membrane immunophenotyping should be used for the           Poor Response To Chemotherapy and Short
confirmation, diagnostic clarificaion of atypical           Survival
leukemias. In this study, 40 cases of acute leukemias       Azza Mahmoud Kamel, M.D.*; Nahla El-Sharkawy,
were classified by morphological and cytochemical           M.D.*; Zekri Khaled Zekri, M.D.**; Hussein Khaled,
stains e.g. MPO, PAS, Acid esterase and phosphatase.        M.D.**; Mohamad Ahmad Al-Akkad, M.D.*** And
For cases were the cytochemical findings were               Mohamad Tawfik Hassan, M.D.***
uninformative or equivocal immunological evaluation         The Departments of Clinical Pathology*, Medical
were      done,     intracellular     and      membrane     Oncology**, National Cancer Institute, Cairo
immunophenotyping were studied, details will be             University and Internal Medicine***, Faculty of
discussed later.                                            Medicine, Zagazig University.
                                                            J Egypt Nat. Cancer Inst. 13(1): 19-26, 2001
9.10. The Expression of Bcl-2 and Bax
Proteins and Their Clinical Relevance in ALL                We have analyzed by immunocytochemistry (ICC) the
and CLL Patients                                            frequency of p53 protein expression in 47 cases of
Hadir A. El-Mahallawy, M.D.*; Nayera H.                     chronic lymphocytic leukemia to assess the
El-Shakankiry, M.D.*; Saad El-Guindy, Ph.D.**;              relationship between p53 and the disease stage as well
Sherine El-Maghraby, M.Sc.*; Sohair Abd El-Latif,           as the possible impact of this protein on the response to
M.D. And Inas El-Attar, Ph.D.***                            treatment and survival. Of these, 42 samples were valid
The Departments of Clinical Pathology*, Tumor               for interpretation. The overall frequency of p53 protein
Biology** and Epidemiology & Biostatistics***,              positivity in CLL was 16.7% (7 of 42 cases). The
National Cancer Institute, Cairo University.                percentage of p53-positive cells increased with disease
J Egypt Nat. Cancer Inst. 13(1): 35-42, 2001                progression (25±7% in stage 3 vs. 10.3±6.2% in stage
                                                            1+2). This difference, however, did not reach statistical
The products of the bcl-2 gene prolong survival of          significance. A significantly poorer response to therapy
lymphohemopoietic cells by inhibition of programmed         was observed in p53-positive compared to
cell death, whereas, bax has an antagonistic role against   p53-negative patients. Three out of 7 p53 (+ve) versus
the function of bcl-2. In the present study, we have        37 out of 35 p53 (-ve) patients showed a response to
examined the expression of bcl-2 and bax proteins in        therapy (p=0.05). The overall survival rate was
cases of ALL and CLL aiming to establish if                 significantly shorter in patients with p53 protein
measurement of expression of these proteins could be        expression (p=0.024). The results of this study indicate
of prognostic relevance. Using quantitative western         that in CLL the expression of the p53 protein is
blotting (WB) and immunocytochemistry (IC) we               associated with poor response to chemotherapy and
determined the level of expression of bcl-2 and bax         short survival.
proteins in 42 and 20 newly diagnosed cases of ALL
and CLL, respectively. Ten healthy individuals were         9.12. Frequency and Clinical Relevance of
taken as control for bcl-2 and bax expression by IC and     TEL-AML1 Fusion Gene in Childhood Acute
WB. By WB, bcl-2 protein was detectable in 39/42 of         Lymphoblastic Leukemia in Egypt
ALL cases and all CLL cases studied. Bax protein            Heba M. Shaker, M.D.; Iman A. Sidhom, M.D.* And
expression using WB revealed detectable levels in           Inas A. El-Attar, Ph.D.**
37/42 ALL cases and all CLL cases studied. The level        The Departments of Clinical Pathology, Pediatric

                                                                                                      9. Leukemia

Oncology*      and     Biomedical       Statistics &       hallucinations, facial palsy, macular edema or
Epidemiology**, National Cancer Institute, Cairo           encephalitis p=0.001. Conclusion: Thus, we report the
University.                                                use of HHV-6 antigenemia for the detection of active
J Egypt Nat. Cancer Inst. 13(1): 9-18, 2001                infection of this virus and that HHV-6 has a
                                                           myelosuppressive effect and we confirm that it is a
TEL-AML1 fusion gene, resulting from 12;21 chro-           CNS pathogen in allogeneic SCT recipients.
mosomal translocation, is believed to be the most
common molecular genetic abnormality in childhood          9.14. Level and onset of CMV-PP65
acute lymphoblastic leukemia (ALL). This study has         Antigenemia As Determinants of Risk For
been conducted to investigate the frequency of this        CMV-Related Complications in Stem Cell
fusion gene in Egyptian children suffering from ALL        Transplant Recipients
and to point out the different laboratory and clinical     Hadir El-Mahallawy, M.D.1; Fatma Salah, M.D.4;
features associated with this anomaly, as well as the      Nahla El-Sharkawy, M.D.1; Alaa El-Haddad, M.D.2;
response of positive cases to therapy. The status of       Inas El-Attar, Ph.D.3; Omar Fahmy, M.D5.; Hossam
TEL-AML1 fusion gene was determined by the reverse         Kamel, M.D.2 And Azza Kamel, M.D.1
transcriptase polymerase chain reaction (RTPCR) in 81      Clinical Pathology Department, Medical Oncology1,
children with ALL, 69 newly diagnosed, and 12 in           Department, Biostatistics        and Epidemiology,2,
relapse. Of the newly diagnosed cases, 7 were positive     Department, National Cancer Institute, Cairo
for TEL-AMLI fusion gene (10.14% of all ALL cases          University3, Research Institute of Ophthalmology4, and
studied, 11.67% of precursor B ALL), as well as three      Department of Medicine, Ain Shams University5.
out of the 12 cases (25% g all ALL cases, 30% of           J Egypt Nat. Cancer Inst 13(4): 259-266, 2001
BCP - ALL) in relapse. All positive cases belonged to
the precursor Blineage, showed an age peak between 3       This study was carried out to investigate if quantifica-
and 6 years, had non-hyperdiploid DNA content and no       tion of CMV antigen can be used to risk adapt the
CNS infiltration. Most of the positive cases had total     prophylaxis for CMV infection in stem cell transplant
leukocytic counts below 50 x 109/l, myeloid marker         recipients in order to reduce the cost and the number of
co-expression and good response to induction therapy.      patients who are exposed to ganciclovir. Assay for
In conclusion, TEL-AML1 fusion gene identifies a           CMV early antigen pp65 on circulating leukocytes was
subset of pediatric acute lymphoblastic leukemia           used to monitor early CMV infection in 54 consecutive
associated with a number of clinical and laboratory        patients undergoing peripheral blood stem cell
markers of good prognosis and should thus be included      transplantation (PBSCT). A total of 201 samples were
in routine molecular workup of acute leukemias to          examined for CMV antigen in peripheral blood
confirm its impact on clinical outcome and to design       leukocytes by the slide method and 40 samples were
suitable therapeutic regimens.                             additionally investigated by flow cytometry. Of 54
                                                           patients examined, twenty five patients (46%) were
9.13. Clinical Relevance of Quantitative HHV-6             positive for CMV pp65 antigen. Eleven of these
Antigen Detection in Leukocytes of Allogeneic              patients developed CMV disease following PBSCT.
Stem Cell Transplant Recipients                            Patients who suffered from CMV disease showed
Hadir A. El-Mahallawy, M.D.; Inas El-Attar, Ph.D.*;        statistically significantly higher OT levels (p=0.003),
Raafat Abd-El-Fattah, M.Sc.** And Hossam Kamel,            higher frequency of complications (p=0.008) and
M.D.**                                                     unfavorable outcome (p=0.002). Patients who were
The Departments of Clinical Pathology, Epidemiology        positive for CMV antigen had a 4.5 times higher risk to
& Biostatistics* and Medical Oncology**, National          develop GVHD. In multivariate analysis, we found that
Cancer Institute, Cairo University.                        CMV pp65 antigen positivity was the only factor,
J Egypt Nat. Cancer Inst. 13(4): 297-301, 2001             which was independently related to GVHD. On
                                                           comparing the detection of antigenemia by the slide
Background: Human herpesvirus-6 (HHV-6) DNA has            method to that by flow cytometry, the slide method
been detected in 70-100% of allogeneic bone marrow         was statistically significantly more sensitive in
transplant and PB stein cell transplant (SCT) patients     diagnosing patients who manifested with CMV-related
few weeks after transplantation using the nested PCR       disease (p=0.01). Thus, antigenemia can be used to
technique. But this may indicate latent HHV-6              assign PBSCT recipients at risk for CMV disease and
infection. Patients and methods: To evaluate the           risk adapted prophylaxis for CMV can be carried out
potential role of the antigenemia assay in the diagnosis   according to the level of CMV positive leukocytes.
of active infection of HHV-6, we subsequently
analyzed PB leukocytes of 50 patients following SCT        9.15. Peripheral Blood: The Challenge in
for HHV-6 antigen. Results: Eighteen patients (36%)        Hematopoietic Stem Cell Transplantation
were positive for HHV-6 by the antigenemia assay and       A Fahmy, H K Mahmoud, A Haddad, S El Badawy, A
infection was significantly associated with reduced        Kamel, H Sedky, S Abdel Latif, M Abdel Moaty, R
total leukocytic count (TLC) and thrombocytopenia,         Abdel Fattah, A Sobhy, A Nassar, M El Emary, T Rabie
p=0.02 and <0.001, respectively. Moreover, HHV-6           and N Adel.
infection was encountered in patients who developed        BMT unit, Nasser Institute, Cairo, Egypt
unexplained CNS manifestations, as 7 cases of the          ISH – European African Division Meeting, 9-14
HHV-6 positive group developed persistent headache,        October 2001, Cairo.

Scientific Annual Report 2000-2002

                                                           University of Edinburgh, Edinburgh EH8 9AG. United
Over the past 25 years, the field of hematopoietic stem    Kingdom [F.E.A.S.L.P.]; Clinica Pediatrica, Universita
cell transplantation (HSCT) has shown a tremendous         Milano, Ospedale S. Gerardo, 20052 Monza, Italy
progress, and consequently there has been an               [A.B.]; Centro Infantil de Investigacoes Hematoldgicas
exponential growth of the number of transplanted           D. Boldrini. Rua Dr. Gabriel Porto, 1270, Bardo
patients as well as the number of indications for which    Geraldo, CEP 13 083 210 Campinas, Sau Paulo. Brazil
the transplantation deemed necessary. Compared to          [S. B.]; Haematology Section. University of Chile,
bone marrow (B.M), peripheral blood (PB) is gaining        Hospital del Salvador, Santiago, Chile [M.E.C.S.L.];
more popularity as the main source of stem cells for       Department of Pathology, University of Hong Kong,
both allogeneic & autologous HSCT. In the period           Queen Mary Hospital Compound, Hong Kong
between April 1997 and September 2000, we have             [L.C.C.]; Shanghai Institution of Hematology,
transplanted 291 patients in the BMT center of Nasser      Shanghai Second Medical University, Shanghai. China
Institute; Cairo, Egypt Out of these, 233 patients         [Z.C.]; Department of Cellular Biotechnology and
received an allo-transplant from HLA-identical             Hematology, University La Sapienza of Rome, Rome,
siblings, while the other 58 Patients had an               Italy [G.C.]; Hospital de Apoto Brasilia, Unidade de
auto-transplant. Seventy five percent of our patients      Onco-Hematologia Pediatrica, SGAIN Q.4. CEP 72
were adults, while children represented only 25%. All      620 000 - Brasilia. Brazil [J.C.C.I.Q.M.]; Department
patients (auto- & allo-) received peripheral blood stem    of Pediatric Hematology/Oncology, Xinhua Hospital
cells which were completely unmanipulated. The main        Shanghai Children's Medical Centre, Shanghai Second
indications for transplant were chronic myeloid            Medical University, Shanghai. China [L.I.G.];
leukemia (CML; 35%), acute myeloid leukemia (AML;          Department of Pediatric Oncology, National Cancer
18%), severe aplastic anemia (SAA; 14%), acute             Institute, Cairo University, Kasr El Eini St, Fom
lymphatic leukemia (ALL; 11%), myelodysplastic             El-Khalig, Cairo, Egypt [H. H.]: Division of Pediatrics,
syndrome (MDS) & solid tumors (6% each);                   Hamanomachi Hospital 3-5-27 Maiatru, Chuo-kit
lymphoma (5%) and hemoglobinopathies (5%). The             Fukuoka 810-8539, Japan [E.I.]; Department of
main causes of death were graft vs. host disease           Clinical Pathology, National Cancer Institute, Cairo
(GVHD; 53%); various infections (30%), bleeding            University, Kasr El Eini St. Fom El-Khalig, Cairo,
(7%), and interstitial pneumonitis & veno-occlusive        Egypt [A. M.K.]: Department of Pediatrics and
disease of the liver (5% each). Indeed, the tempo of       Developmental Biology, Postgraduate Medical School,
hematopoietic recovery was quite tempting; for the         Tokyo Medical and Dental University, I-5-45,
allotransplanted group, the mean number of days for        Yushima, Bunkyo-kit, Tokyo 113-8519, Japan [S.M.];
neutrophil and platelet recovery was 14 & 19,              Department of Hygiene and Epidemiology, University
respectively, while for the autotransplanted group, they   of Athens, School of Medicine, 11527 Athens. Greece
were 17 & 22, respectively. When we followed our           [E.P.]: Instauto Nacional de Cancer, Prato Cruz
patients to establish a Kaplan - Meier curve for each      Vermelha, 7o. andar, laboratono de Marcadores
group of diseases, we found that the 3 year                Celulares-Hematologia Cliniea, CEP 20230-130, Rio
disease - free survival was: for CML (transplanted in      de Janeiro. Braid [M.P.D.O.]; Department of
1st chronic phase) 75%, for SAA 63%, for AML 45%,          Paediatrics, The Chinese University, Hong Kong
for ALL 56%, for B-thalassemia 80%, while it was           [P.Y.]; and Institute of Cancer Research, Chester
58% for the whole 291 transplanted patients. In            Beatty Laboratories, London SW3 QJB, United
agreement with the prevailing view, when we                Kingdom [J.L.W.M.F.G.]
compared the incidence of acute & chronic GVHD in          Cancer Research 61: 2542-2546, 2001
this group of patients to our historical BMT group, we
did not find any significant difference from the           Infant acute leukemia (IAL) frequently involves
statistical point of view. In conclusion, the peripheral   breakage and recombination of the MLL gene with one
blood stem cells appear to be a more tempting source       of several potential partner genes. These gene fusions
for autologous & allogeneic HSCT compared to BM            arise in utero and are similar to those found in
stem cells. Moreover, many of the previous worries         leukemias secondary to chemotherapy with inhibitors
regarding their use should be superseded when              of topoisomerase II (topo-II). This has led to the
engraftment, tempo of hematopoietic reconstitution,        hypothesis that in utero exposures to chemicals may
and GVHD are concerned.                                    cause IAL via an effect on topo-II. We report a pilot
                                                           case-control study of IAL across different countries
9.16. Transplacental Chemical Exposure and                 and     ethnic    groups.    Cases    (n=136)      were
Risk of infant Leukemia With MLL Gene                      population-based in most centers. Controls (n=266)
Fusion                                                     were selected from inpatients and outpatients at
Freda E. Alexander, Sherry L. Patheal, Andrea Biondi,      hospitals serving the same populations. MLL
Silvia Brandalise, Maria-Elena Cabrera, Li C. Chan,        rearrangement status was derived by Southern blot
Zhu Chen, Giuseppe Cimino, Jose-Carlos Cordoba,            analysis, and maternal exposure data were obtained by
Long-Jun Gu, Hany Hussein, Eiichi Ishii, Azza M.           interviews using a structured questionnaire. Apart from
Kamel, Silvia Labra, Isis Q. Magalhaes, Shuki              the use of cigarettes and alcohol, very few mothers
Mizutani, Eleni Petridou, Maria Pombo de Oliveira,         reported exposure to known topo-II inhibitors.
Patrick Yuen, Joseph L. Wiemels, and Mel F.                Significant case-control differences were apparent for
Greaves Department of Public Health Science.               ingestion of several groups of drugs, including herbal

                                                                                                       9. Leukemia

medicines and drugs classified as "DNA-damaging,"          Leukemias
and for exposure to pesticides with the last two being     Mostafa I. Aboul-Enein, Randa M. Aboul Fetouh,
largely attributable, respectively, to one nonsteroidal    Khaled M. Aboul Enein, Ahmed A. Shams El Din* and
anti-inflammatory drug, dipyrone, and mosquitocidals       Amal A. Aly Saadallah**
(including Baygon). Elevated odds ratios were              Egypt J Lab Med 13(3): 435-442, 2001
observed for MLL+ve (but not MLL-ve) leukemias (2.31
for DNA-damaging drugs, P=0.03; 5.84 for dipyrone,         This study involved 11 cases of AML, 10 cases of ALL
P=0.001; and 9.68 for mosquitocidals, P=0.003).            and 1 case biphenotypic acute leukemia. All cases were
Although it is unclear at present whether these            subjected to morphological, cytochemical and
particular exposures operate via an effect on topo-II,     immunophenotypic analysis for typing of acute
the data suggest that specific chemical exposures of the   leukemia. At the same time the phagocytic test using
fetus during pregnancy tray cause, MLL gene fusions.       candida albicans was done to all cases and compared
Given the widespread use of dipyrone, Baygon, and          with the other differentiating parameters for the type of
other carbamate-based insecticides in certain settings,    blast. The results showed that occasional myeloblasts
confirmation of these apparent associations is urgently    in every AML case engulfed candida i.e. possessing
required.                                                  early phagocytic function. But the blasts of the
                                                           biphenotypic AL and ALL cases did not show any
9.17. A Suggested Flow Chart For Diagnosing                phagocytic power. The positivity of the phagocytic test
Biphenotypic (Hybrid) Acute Leukemia                       matched that of SBB, Peroxidase & Chloroacetate
Azza M. Kamel, Nahla M. El-Sharkawy, Fayek M.              esterase tests. Hence, it is proposed to use the
Ghaleb*, Heba M. Shaker, Dina A. Yassin, Nayera            phagocytic function as a new parameter to help
Hamdy                                                      differentiation of the myeloblast of AML from the
Clinical Pathology, Departments, National Cancer           lymphoblast of ALL.
Institute, Cairo University and Ophthalmology
Research Institute, Academy of Scientific Research*        9.19. Nonmyeloablative Transplants For
6th Meeting of the European Association of                 Hematological Disorders
Hematology, Frankfurt, Germany, June 2001.                 Mahmoud HK, El Haddad A, Fahmy O, Kamel A, El
                                                           Shakankery N, Mossallam G and Nassar A.
Acute Leukemias are basically classified according to      BMT Units, Nasser institute and NCI, Cairo.
morphological criteria and cytochemical findings into      The 7th Annual Conference of The Egyptian Association
two broad categories of lymphoid (ALL) and non-            of Immunologists, Ein Soukhna, Jan 2001.
lymphoid leukemias (ANLL). After the introduction of
monoclonal antibodies (mAbs), increasing number of         Although the myeloablative conditioning regimen with
reports are appearing in the literature describing cases   allogeneic bone marrow transplantation has been used
expressing both myeloid and lymphoid markers. In the       to treat patients with hematologic malignancies, yet the
present study morphology, cytochemistry, and detailed      regimen related toxicities has limited the use of such
immunophenotyping of 497 newly diagnosed acute             treatment modality to younger and medically tit
leukemia cases were performed. The cases were 382          patients. The objective of this study was to assess the
ALL, which included 302 child (≤16years) and 80            feasibility of replacing the myeloablative conditioning
adult and 115 ANLL patients, which included 63             regimen with a less toxic non-myeloablative
children and 52 adult. Twenty-nine ANLL cases              conditioning for the elderly and medically infirm
expressed lymphoid markers and 41 ALL cases ex-            patients. Between October 2000 and November 2001,
pressed myeloid markers. In few cases, gene                36 patients were enrolled (20 males and 16 females)
rearrangement and TCR protein expression were done.        with the diagnosis of chronic myelocytic leukemia
The incidence of myeloid marker expression in ALL          (n=11), acute myeloid leukemia (n=11), acute
cases were 10.7%; being 8.9% in children and 17.5 %        lymphoblastic leukemia (n=4), myelodysplastic
in adults. In the ANLL group, cases expressing lym-        syndrome (n=3), chronic lymphocytic leukemia (n=2)
phoid markers were 20% being 23.8% in children and         and Fanconi anemia (n=2).The mean age was 34.6
15.4% in adults. Three scoring systems (Hurwitz and        years (range=6-55 yrs). All patients received
Mirro, Catovsky et al and EGIL) were applied to our        fludarabine 30 mg /m2 for 3 days and total body
cases and the phenotype verified. We have suggested a      irradiation with 200 cGy followed by infusion of
flow chart, in which the interpretation of marker          G-CSF mobilized peripheral CD34 +ve cells and post
expression should be clone in context of the original      transplantation         immunosuppression         using
diagnosis being ANLL or ALL, achieved by                   mycophenolate mofetil and cyclosporine A. Chimerism
morphology and detailed cytochemistry. In certain          studies (VNTR) were performed at D28 and D36.
conditions confirmatory tests as TCR protein               Procedure related toxicities were minimal. Early
expression and gene rearrangement may be needed.           engraftment with full donor chimerism was observed in
The flow chart should take in consideration any genetic    16/36 and mixed chimerism in 10/36. Acute GvHD
markers available.                                         more than grade 2 was observed in 4/36 and extensive
                                                           chronic GvHD in 12/36. DLI was required in 9/39
9.18. The Use of The Phagocytosis Test As A                patients (CML 8/9 and AML 1/9). The disease was
Differentiating  Parameter   Between    the                controlled in 5/9 and progression was diagnosed in 4/9.
Mylloblast and Lymphoblast in Acute                        GvHD occurred in 3/9 following DLI (grade 1 in 2/9

Scientific Annual Report 2000-2002

and grade 3 in 1/9). The OS was 21/36 and DFS was          Thrombosis & Haemostasis. ISTH Paris 2001, July
17/36 at a median observation of 5 months,                 6-12.
non-myeloablative HSCT is the first step in
re-evaluation of how to perform allo-HSCT in the           Graft Vs. Host Disease (GVHD) is still one of the
future and appears to be safe and minimally toxic even     leading causes of morbidity and mortality following
in patients who would normally be excluded from            allogeneic hematopoietic stern cell transplantation. A
allo-HSCT. However, longer follow up and more cases        concern is still going on regarding the incidence of
are needed to reach a firm conclusion.                     chronic GVHD in patients allografted with peripheral
                                                           blood stem cells (PBSCs). We have already reported
9.20. Proliferative and Prognostic Significance            the feasibility of allogeneic PBSCT in 18 heavily
of DNA Ploidy, Nucleolar organizer Regions                 pre-transfused patients with severe aplastic anemia
and Morphometric Analysis Using Image                      (SAA), and showed that the incidence of acute GVIID
Cytometry in Pediatric Acute Leukemias                     is not increased, if not even less, as compared to
Mostafa Aboul-Enein; Somya A.K. El- Hosseiny*;             patients receiving marrow cells. In the current study,
Sherif A. Aboul-Naga**; Bahaa B. Ghannam***                we are reporting on the incidence of chronic GVHD in
Nahla M. Leheta****; Khaled M. Aboul Enein; Randa          the same cohort of patients. In the period between July
M. Aboul-Fetouh and Somaya M. Abdel - Monem                1997 and November 1998, 11 males and 7 females
Dept. Clinical Pathology, NCI, Cairo University;           (mean age 19) received Filgrastim mobilized PBSCs
*Dept. Pathology, NCI, Cairo University; **Dept.           from their HLA-identical siblings, and all were
Pediatric Oncology, NCI, Cairo University; ***Dept.        conditioned with Cyclophosphamide + ATG. The mean
Pathology, El Azher University; *Dept. Clinical            number of infused unmanipulated CD34+ve cells was
Pathology, Cairo University                                7.5x106/kg body weight. All patients received GVHD
Egypt. J. Lab. Med. 13(2): 417-432, 2001                   anaphylaxis using Methotrexate + Cyclosporine. The
                                                           tempo of hematopoietic reconstitution was rapid (mean
This work, involved the study of DNA ploidy,               number of days for neutrophil and platelet recovery
Ag- NOR and morphometric analysis by Image                 was 11 and 15, respectively), and no single early or late
Analyzer in pediatric acute leukemia. Forty five           graft failure was reported. The main causes of death
subjects were studied (35 de novo pediatric acute          included intracranial hemorrhage (n=1), CMV
leukemia patients and 10 healthy children controls).       infection (n=1), interstitial pneumonitis (n=1), sepsis
Twenty-three cases had ALL and 12 had AML. The             (n=1), late case infectious meningitis (n=1) and acute
scatter of ploidy showed similarity between ALL and        GVHD (n=2). No other cases of acute GVHD was
AML cases. Hence, ploidy was not affected by the type      observed. During a follow-up period of up to 36
of acute leukemias. Aneuploidy occurred in 31.4% and       months, curly one patient (5.5%) developed chronic
tetraploidy in 5.7%, while most of the cases (62.9%)       GVHD, which was limited and well controlled under
were similar to the normal control i.e. diploid pattern.   immunosuppressive therapy. In conclusion, our data
Ag-NOR count was significantly higher in ALL (3.8          suggest that in patients with SAA, allogeneic PBSCT is
±1.3) and AML (3.2±1.4) than in controls (l.9±0.09).       not only associated with rapid engraftment but also
Bone marrow morphometric mean nuclear area was             with decreased incidence of both acute and chronic
higher in both ALL patients (52.5±17.0) and AML            GVHD.
patients (85.7 ±35.0) as compared to normal controls
(12.7±2.3). As regards prognosis DNA ploidy pattern        9.22. The Decisive Role of Bone Marrow
showed no relation to prognosis, however Ag-NOR            Trephine Biopsy in Cytopenias of Hemopoetic
and morphometric mean nuclear area (MNA) parallel          Malignancies
prognosis. MNA is considered a survival parameter as       Aboul-Enein M. I.*, Khoirshid A. M.*, Mokktar N. M.
it was less in patients attaining complete remission.      *, Aboul-Enein K. M. P`, Kholoussi N. M. **, Moussa
Also, if characterizes together with Ag-NOR count L1       H. S.*, and Zaher A. E.*
and L2 subtypes of ALL. Thus, Image Cell Analyser          *Clinical Pathology & Pathology Departments, NCI,
not only shows high capability and sensitivity to detect   Cairo University, **Human Genetics Department,
small aneuploid populations, but also proved to be the     National Research Centre
most recent approach for standardized Ag-NOR and           Egypt. J. Lab. Med. 13(3): 435-442, 2001.
morphometric analysis. Ag-NOR and MNA are
promising markers to monitor prognosis of malignant        Objectives: This study involved the evaluation and
calls.                                                     decisive diagnostic role of BM trephine biopsy in the
                                                           etiology of cytopenias of hemopoietic malignancy.
9.21. Acute and Chronic Graft Versus Host                  Design: Selective study design. Patients: Some cases of
Disease Following Primary Transplantation of               failed aspiration subjected to BM trephine biopsy
Allogeneic Peripheral Blood Stem Cells in                  showed normal cellularity or hypercellularity were
Patients With Severe Aplastic Anemia                       excluded when not cytopenic. This work included 104
O.A. Fahmy, H.K. Mahmoud*, A. E. Haddad*, A.               cytopenic cases: 28 Aplastic Anemia (AA), 24
Kamel*, M. Nazih, A. Sobhy*, T.S. Rabie*, M.A.             Hodgkin's disease (HD), 23 non Hodgkin's lymphoma
Hassaballa*.                                               (NHL), 1 myelofibrosis (MF), 1 myelosclerosis (MS),
BMT Team, Nasser Institute, Cairo, Egypt.                  11 rnyelodysplastic syndrome (MDS), 8 hairy cell
XVIII Congress The International Society on                leukemia (I-ICE), 3 chronic myeloid leukemia with

                                                                                                          9. Leukemia

myelofibrosis (CMLMF), 2 chronic lymphocytic
leukemia (CLL), 1 metastasis, and 2 hypersplenism.            In this study 80 patients suffering from acute leukemia,
Setting: National Cancer Institute, Cairo University.         chronic leukemia, lymphoma and solid tumors were
Intervention: Of the 104 cases only cytopenic 83 cases        subjected to CBC, BM examination, phagocytosis. C3
were studied by BM trephine biopsy with different             and protein electrophoresis. The aim is to detect if the
stains, (hematoxlin and eosin (H&E), Giemsa,                  neutrophils in malignant disorders are defective only in
Reticulin,     Toluidine      blue     O     (TBO)     and    number or in number and function and to detect
immunohistochemistry (IHC) to explore the etiologic           immunologic status in different levels of neurtopenia.
diagnosis. Results: Cases with AA showed mostly fat           The patients wew grouped as follows: group I Acute
spaces, some showed rare lymphocytes or occasionally          leukemia (28 ALL& 6 AML): 34 cases. Group II
tissue Mast Cells. In HD, G of 24 from the studied            Chronic leukemia (8 CLL&4CML): 12 cases. Group
cases showed Reed-Sternberg (R.S.) or Hodgkin (H)             III Lymphoma (2 HD & 12 NHL): 14 cases. Group IV
cells and stained with CD IS and CD 30 11-IC.                 solid tumors: 20 cases. Group V: 10 normal controls.
Cytopenia associated with hypocellularity but showed          Results revealed that in neutropenia in malignancy the
the characteristic HD cells. NHL were classified into         neutrophil had an increased phagocytic power to a
nodular, interstitial, or diffuse infiltration. B or T-cell   remarkable degree when compared with non-
origin by II4C was diagnostic of NHL associated with          neutropenic patients or normal controls. Also, the Beta
cytopenias, mostly due to chemotherapy/radiotherapy           and Gamma globulis in protein electrophoresis and C3
effect. Myelofibrosis case studied showed that                were higher in neutropenic patients when compared
cytopenia revealed extensive marrow fibrosis which            with non-neutropenic patients or normal controls.
was evidently stained with Reticulin stain.                   Complement C4 did not show any important changes.
Myelosclerosis cytopenic case showed extensive                CRP changes, decreas in total protein and albumin and
trabicular proliferation and thicking with scanty small       decrease in immunoglobulins specially IGM and IGA
areas left for marrow. MDS cases with cytopenias              were related to malignancy is compensated by
showed varieties of dysplasia in the trephine biopsy.         increased phagocytic power, and Beta and Gamma
Uni and multilobular megakaryocytes clustered                 globulins. These compensatory levels of defense
paratrabicular were seen. Abnormal Localisation of            parameters in neutropenic are possibly enough to
Immature Precursors (ALIP) was noted in some cases            prevent infection in all cases studied.
(refractory anemia with excess blast RAEB). One case
with BM aspiration showed 5% blasts, the trephine             9.24. The Role of Scanning Electron
biopsy revealed many blast nests (ALIP) which                 Microscopy in The Diagnosis of Some
changed the diagnosis to acute myeloid leukaemia              Hematological Malignancies
(AML). As regards to HCL, BM trephine biopsy was              Amira Khorshed,*, Naglaa M. Kholoussi,**,
decisive in showing hairy cells, honey comb                   Aboul-Enein I. M.*, Zaki A. M. ** and Aboul-Enein
appearance in these cytopenic cases. In CML/MF, the           M.K.*.
count was close to normal and BM trephine biopsy              *Clinical Pathology Dept, National Cancer Institute,
showed remarkable MI, as one of the complications of          Cairo University, **Human Genetics Dept, National
CML. The hemogram in CLt_ cases showed anemia,                Research Center.
neutropenia and till omhocytopenia though evident             Egypt J Hematol 26(2), 331, 2001
lymphocytosis. These cases were late (treated for
years). BM trephine biopsy showed very scanty                 Accurate categorization of leukemic cells in clinical
granulocytes and increased tissue mast cells as               hematology depends on the French-American-British
evidenced by Toluidine blue-O. Metastasis case slowed         classification. Recently the WHO proposed some
marked cytopenia and BM trephine biopsy revealed              modifications; the morphology of leukemic cells is one
unilateral iliac metastasis. Two hypersplenism cases          of the important criteria in this classification. In this
with pancytopenia .showed evident BM hyperplasia in           study, we distinguished leukemic cells under the
the trephine biopsy. Conclusion: BM trephine biopsy           scanning electron microscopy (SEM) in 40 cases of
should be done in: 1-dry tap 2-most hemopoietic               different hematological malignancies. The surface
malignancies especially with cytopenias, unless BM            topography of different leukemic cells, the size of cells
aspiration is sufficient as in acute and chronic              and the surface changes were identified anal described
leukemias and hypersplenism. 3-bilateral iliac trephine       for each hematological disease. These parameters were
biopsy in essential in HD, NHL, and metastases to             compared with the parameters seen by the transmission
avoid missing cases with uneven distribution                  electron microscope (TEM). This was a complete
4-supplementary IIIC (for early invasion), Reticulin          descriptive study for the different leukemic cells.
and T130 stains give important descriptive features.
                                                              9.25. MDR-1 Gene Expression in Egyptian
9.23. Study on The Compensatory Activation                    Leukaemia: Correlation With Phenotype and
of The Neutrophils and Some Defense                           Treatment Outcome
Mechanisms in Neutropenia of Malignancy                       Abdel-Rabman Zekri**, Moushira M El Saifi*, Maha
Mostafa I. Aboul Enein*, Omainia A. Gohar*, Walaa             Saleh*, Iman Attia***, and Sabreen Abd Allah**
A. Cag*, Nehad E. Moussa,Khaled M. Aboul.Enein*,              *Clinical Pathology Dept, **Cancer Biology Dpt, and
Nahed Abdel Wahab* and Hisham M. Shahin*                      ***Pediatric Oncology Dept, National Cancer Institute,
Egypt J Hematol 26(1): 31, 2001                               Cairo University.

Scientific Annual Report 2000-2002

Egypt J Lab Med 13(1): 35-46, 2001                         responders were close to normal level. GD95 (cells
                                                           going to apoptosis) control normal (7.04±1.0). As
One important mechanism of drug resistance in acute        regards CML before treatment (3.1±2.4) and after
leukemias is over-expression of the multi-drug             treatment responders (1.6±16.9) and non responders
resistance (MDRI) gene that encodes a 170-KDa              (11.0 ±5.4). Before treatment there was significant low
membrane protein called P-glycoprotein. To estimate        level of cells going to apoptosis (CD95+ve) as
the incidence and role of MDRI gene expression in          compared to normal control level. After treatment,
patients with acute leukemia, we investigated the          there was quite high CD95+ve cells in the responders
expression of MDR-l by using die RT-PCR method in          (many cells going in apoptosis). The non responders
blast cells from 43 patients with de novo acute            shows prominent: increase in number of CD95+ve cells
leukemia. We found a high frequency of MDR-I gene          than the responders but still much higher than before
expression: 17 out of 32 with de novo ALL and 3 of II      treatment or controls. As regards CLL before treatment
with de novo ANLL, were MDR-1 in RNA -positive.            CD95 level is significantly less than normal controls
No correlation between MDR-1 gene expression and           (3.5±1.9), after treatment responders were very high as
clinical data as patient’s age, sex, WBC count at          compared to control and before treatment (18.5±7.4)
presentation, Hb level, PB or BM blasts or                 and the same for non responders (18.7 ±10.6) ( ? due to
immunophenotype. Multivariate analysis using logistic      marked increase in cells ). As regards DLL before
regression revealed a lower CR rate among MDR-1            treatment CD95 level was (3.3 ±2.5) and after
positive cases as compared to those without detectable     treatment responders (12.6±3.3) and non responders
expression in the ALL group (P=0.02). These results        were (14.6 ±9.2) i.e. significantly lower in pre-
suggest that MDR-1 gene expression can be used as          treatment and higher after treatment than normal
prognostic factor and may be helpful in determining        controls (similar CML and CLL). The still high level of
chemotherapeutic protocols for patients with acute         CD95 in non responders may be due to increase in the
leukemia.                                                  number of cells i.e. increase in apoptotic and non
                                                           apoptotic cells s suggested that Telomerase activity
9.26. The Role of Telomerase Activity and                  could be used as tumor marker for diagnostic and
CD95 in The Pathogenesis of Some                           prognostic purpose
Hemopoietic Malignancies and Their Relation
To Therapeutic Response                                    9.27. Light and EM Studies on The Phagoytic
Aboul Enein M.I.; Khorshid A.; Mansour M.T.; Aboul         and Chemotactic Activities in Normal Myeloid
Enein K.M.; Leheta N.M * ; Khalafalla O. A. *and           Series, Leukemias, Lymphomas and Solid
Raslan H.N.                                                Tumors
Dept of Clinical path. NCI, Cairo University, *Dept of     Mostafa I. About-Enein, Amira M. Khorshid, Sohair A.
Clinical path. Faculty of Medicine, Cairo University.      Isa, Samia Y. Akel, Zakaria A. Alyan, Amr H. Ziada,
Egypt J Hematol 26(4): 785, 2001                           Farida Gadallah, Mohamed A. Taha*
                                                           Dept. Clin. Path., NCI, Cairo University, * Military
This study involved the potential of apoptosis by assay    Medical Academy.
of Telomerase and CD95 in the pre-treatment stages of      Egypt J Hematol 26(4): 785, 2001
CLL, CML and DLL. The cases studied were 50 newly
diagnosed patients, 20 CML, 10 CLL and 20 DLL and          The study was conducted on 67 cases from the
50 cases following treatment were 25 CML ( 20              outpatient Clinics, NCI, Cairo University: 10 controls
responders & 5 non responders), 10 CLL ( 7                 (were anemia for investigation), 10 ML cases, 12 ALL
responders & 3 non responders ) and 15 DLL (11             cases, 3 CML cases, 4 CLL, 7 HD, 15 NHL, 2 MM, 1
responders & 4 non responders). Also 20 normal             MDS, 2 neuroblastoma, l Rhabdomyosarcoma. Bone
subjects were studied as control. All cases were studied   marrows collected from these patients and the normal
clinically, CBC, BM, assessment of Telomerase              controls were subjected to the following studies: (a)
activity by PCR and ELISA and CD95 % by                    Phagocytosis of Candida albicans. (b) Phagocytosis of
Flowcytometry. Telomerase Activity (normal controls        NBT. (c) Chemotaxis. The marrows in [a & b] were
0.09 ±0.06) As regards CML before treatment (1.2±0.        examined by the light and electron microscope. The
7), after treatment responders (0.09±0.05) and non         results are summarized as follows: In normal controls:
responders (1.02±0.6). Before treatment and non            Phagocytosis of candida & NBT showed early activity
responders were significantly higher than normal           in the promyelocyte stage than myelocyte, juvenile,
controls but the responders were within normal control.    and band and segmented. The activity increased as
As regards CLL before treatment (0.6±0.3) and after        maturation proceeds. Blast could not be assessed due to
treatment: responders (0.07±0.03) and non responders       their extreme sparcity in benign cases. Chemotaxis
(0.2±0.01). Before treatment and non responders were       started at promyelocytes and increased as maturation
significantly higher than control normal but the           proceeds. Direct chemotaxis range between 20-35
responders were close to normal control level. As          [mean 28.2±5.5]. Electron microscopy showed
regards DLL before treatment (1.4±1.1) and after           phagocytosis of candida in different stages of the
treatment: responders (0.1±0.04) and non responders        granulocytic series. Secondary ultrastructural changes
(1.9±0.6). Before treatment and non responders were        were decrease in the granules contents, sometimes
significantly higher (frequent primitive cells) than all   leaving vacuoles after emptying their contents, also
studied groups as well as the normal controls but the      large glycogen masses were noticed. In acute leukemia

                                                                                                         9. Leukemia

cases: phagocytosis of candida & NBT showed                  with disseminated tumors) in addition to 20 normal
engulfment by the myeloblasts of AML but riot the            controls. The results showed VWF: patients with
lymphoblasts of ALL. Chemotaxis were 3 times more            disseminated bladder had significantly higher levels of
in AML (84±4.5] than in normal [28.2±5.5] and in             VWF compared with those with primary disease as
ALL [29±2.5]. In Chronic leukemia cases:                     well as control group (153.15±11.73 vs. l25±16.99,
phagocytosis of candida showed increased activity in         50.45±8.15 respectively) p=0.0001 with higher levels
all granulocytic series in CML as compared to normal         in grade III compared with grade II. Thrombomodulin:
and CLL. In NBT; the phagocytic activity was slightly        Patients with disseminated disease have statistically
higher in CML than normal but in CLL, it was                 significant higher levels than those with primary
generally below normal in all granulocytic series.           disease and the control group 5.84±0.55 vs. 4.9±1.47,
Direct chemotaxis in CML was nearly double                   4.11±0.35 respectively) p=0.0001 significantly higher
[65.5±0.7] the normal value [28.2±5.5] and CLL               levels were present in those with squamous cell
[25.6±4]. In HD: Phagocytosis of candida showed              carcinoma compared with transitional carcinoma.
increased activity in promyelocytes, myelo and               PG12: the levels pf PG12 was statistically higher in
meta-myelo, than in normal while the band and mature         patients with primary bladder cancer compared with
segmented were similar to normal. In NBT, the                disseminated disease as well as the control group
phagocytic activity was decreased in the pro-myelo,          (14449±1872.02       vs.     32.5±41.2,      12.9±10.39
myelo, and metamyelocytes than, normal. Similar,             respectively) p=0.0001 with higher levels in more
activity in the band, while below normal in the mature       advanced and positive nodes. In conclusion:
segmented. Direct chemotaxis was [52.2±3.5] nearly           Assessment of these factors may potentially denote
double the normal value [28.2±5.5]. In NHL:                  disease progression. Further studies are still needed in
Phagocytosis of candida and NBT showed higher                larger number of patients with longer follow up.
phagocytic activity, from the promyelocytes down as
maturation progress till the mature segmented, as            9.29. Impact of Graft Components on
compared to normal. Direct chemotaxis showed                 Engraftment Kinetics in PBSC Transplantation
[58±4.2] nearly, double the normal value [28.2±5.5]. In      Azza Kamel, Nahla El-Sharkawy, Hossam kamel,
miscellaneous disorders: The phagocytic activity             Mohammed Raafat Khalaf, Alaa El-Haddad, Omer
(candida & chemotactic power, though yielded                 Fahmy, Doaa Sayed.
interesting results, yet comments cannot be deduced to       NCI, Cairo University, South Egypt Cancer institute,
be applied on similar conditions because of the small        Assiut University, Faculty of Medicine, Cairo
sample size. It is concluded that phagocytosis occurred      University, Cairo, Assiut, Egypt.
in the myeloblasts of ANLL but not in the                    29th World congress of the international society of
lymphoblasts of ALL. This can be used for                    hematology, Soeul, South Korea, Aug. 2002
differentiation from each other. Comparison between
malignant cases showed that phagocytic power was             Objectives: To evaluate, probably for the first time, the
highest in ANLL, CML, NHL high in HD and lowest              impact of CD34 subsets on engraftment kinetics in
in ALL and CLL.                                              allogeneic PBSC transplantation. Methods: The PBSC
                                                             graft components were analysed in 40 cases for the
9.28. Endothelial Release Markers Denoting                   absolute count/kg of total CD34- and its following
Progression in Bilharzial Related Bladder                    subsets: DR- & +, CD71- & +, CD38- & +, CD 33- &
Cancer in Egypt                                              + and CD6l -& +, Time to neutrophil count of ≥0.5 &
Mustafa I. Abou1-Enein*; Nahed Abdel Wahab*;                 ≥1x109L and to platelet count of ≥20 & ≥50x109L was
Khaled Aboul-Enein*; Hussein E. Gaballah**;                  reported. The median value for each parameter was
Hisham shahin*; and Inas El-Attar***                         used to discriminate rapid from slow engraftment.
*Clinical Pathology, **Medical Oncology and                  Results: Two patients died before engraftment and
***Biostatistics and Epidemiology Departments,               were excluded. Fair correlation was encountered
National Cancer Institute Cairo University                   between        CD34+/DR-,         CD34-/CD38-         and
Egypt J Hematol 26(1): 113, 2001                             CD34+:CD71- on one side and platelet engraftment on
                                                             the other (r=0.39/0.35, 0.37/0.35 and 0.36/0.35
Background cancer metastases involves multiple,              respectively). Five parameters showed significant
sequential complex, interacting, and interdependent          predictive power of early neutrophil engraftment. The
steps in which tumor cells undergo complex interaction       best was CD 34+/DR- cell dose; it showed an 85%
with the host platelets and vascular endothelium prior       probability of early engraftment at a Cutoff dose of
to establishing a secondary colony. Components of the        1.29 x 106/kg (P<0.003). The next best were
blood clotting pathway may contribute to metastases by       CD34+/CD38- and CD34-/CD61-; they showed a 75%
trapping cells in capillaries or by facilitating adhesions   probability of early engraftment at cutoff doses of 3.94
of cells to capillary walls. The endothelium is an           and 6.78x106/kg (P=0.011 & 0.03 respectively). The
important source of the VWF, prostacyclin, fibronectin       other 2 parameters were total , CD34± and
and thrombomodulin. This study was intended to               CD34+/CD71-cell dose; they showed a 70%
access the plasma levels of VWF, prostacyclin, and           probability of early engraftment at Cutoff doses of 8.95
thrombomodulin in patients with primary and                  and 4.32x106/kg (P=0.026 & 0.043 respectively). None
metastatic bilharzial related bladder cancer. Results this   of the tested subsets was a significant predictor of early
study involved 40 patients (20 primary tumor and 20          platelet engraftment. Conclusion: In allogeneic PBSC

Scientific Annual Report 2000-2002

transplantation the best predictor of early neutrophil        engraftment from other causes. The recent method of
engraftment is the absolute CD34+/DR- cell dose               transplantation with nonmyeloablative conditioning
followed by CD34-/CD38- and CD34--CD61 and last               "minitransplant" can he only performed under the
total CD34- and CD34-/CD71- cell dose. No predictor           guidance of the status of chimerism in the recipient.
of early platelet engraftment was encountered: yet the        However, a lot of controversies exist. First MC per se
engraftment showed fair correlation to CD34-/DR-,             is not always associated with a higher incidence of
CD34-/CD38- and CD34+/CD71- cell dose.                        relapse: the argument is that detection of chimerism is
                                                              not leukemia specilic (3). Second relapse is net always
9.30. Detection of Chimerism in Allogeneic                    preceded by an increasing MC. Apparently the
BMT: Methodology and Clinical Significance                    sensitivity of the method used and the frequency of
Azza Kamel                                                    testing for MC are behind these controversies.
NCI, Cairo University                                         Quantitative estimation, lineage specific detection and
The XVI Meeting of The International Society of               more frequent monitoring of MC are suggested to solve
Hematology, European and African Division, Cairo,             these controversies (4). In conclusion detection of MC
Egypt, Jan 2002                                               is currently one of the routine investigations performed
                                                              in allogeneic BMT patients. Sensitive and accurate
In medicine, the term chimera is used to designate an         quantitative methods for detection of MC are now
Organism whose body contains cell populations                 available. MC detection is very helpful in DD of failure
derived from different individuals of the same or             of engraftment and GVHD. MC per se does not
different species occurring spontaneously or produced         necessarily anticipate the occurrence of relapse, but
artificially (1). In successful allogeneic BMT the donor      rather an increasing MC. Close monitoring for MC
hematopoietic cells would totally replace the recipients,     preferentially in a lineage specific manner is the best
a condition referred to as complete chimerism (CC).           method for early anticipation of relapse.
However, a low percentage of recipient hemopoietic
cells may persist, which is called mixed chimerism            9.31. Karyotype and Immunophenotype in
(MC). The Presence of residual recipient cells always         Pediatric Acute Lymphoblastic Leukemia;
carried the risk that these cells might proliferate           Impact on Clinical Presentation and Duration
overcrowding the donor cells with ultimate relapse or         of First Remission
autologous reconstitution. Accordingly, the interest in       Fayza     Hamouda,       Farida      Gaballah,     Nahla
detection of MC started from the very begining of             El-Sharkawy, Ashraf Khairy*, Azza H. El-Sissy, Azza
allogeneic BMT. Historically, markers as red cell             Kamel.
phenotypes, Ig allotypes and karyotyping were used.           Clinical pathology department, (*) Pediatric oncology
Currently molecular techniques are the tool of choice         unit, National Cancer Institute, Cairo University.
in detection of chimerism. Three different approaches         XVIth Meeting of the International Society of
are used for Detection of MC: (1) In sex matched              Hematology, European & African Division, Jan 5-9,
transplants with male patient and female donor,               202, Cairo, Egypt.
characterization of Y chromosome specific sequences
is performed either by FISH technique, PCR and/or             This is a retrospective analysis of 40 pediatric ALL
RFLP. (2) In recipients with disease marker, detection        patients who presented to the pediatric unit, National
of MRD is the method of choice: this will detect only         Cancer Institute, Cairo University. The studied patients
malignant cells and not all recipient cells. The protoype     were divided into four groups based on cell surface
of this approach is the detection of BCR/ABL chimeric         markers. The first group is the Pro B ALL represented
gene in CML cases. (3) In cases where neither of these        by 5/40 (12.5%), their mean age was 5.4 years with a
approoaches is appthse use vanumhcr tandem repeats            M/F 1.5:1, the mean total leukocytic count was 66.6,
(VNTRs), also called microsatellites or short tandem          massive organomegaly was encountered in 3/5 (60%).
repeats (STR), also called microsatallites, is the method     The normal karyotype was present in 4/5 (80%) with a
of choice. These are tandemly repeated sequences              mean first complete remission (CR1) 10.5 months. The
(8-80 bases in VNTRs and 2-7 in STR) present                  second Group was the CALL representing 13/40
throughout the genome. Polymorphism results from              (32.5%), their mean age was 9.8 years, with M/F 1.4:1,
variation in the number of copies of the tandemly             the mean total leukocytic count was 77.046, massive
repeated motif at each sue or locus. They can be              organomegaly was present in 9/13 (69.23%), normal
detected by RFLP or PCR and quantitative estimation           karyotype was present in 2/13 (15.38%) with mean
of STR is the most sensitive method currently                 CR1=16.5 months, hyperdiploid karyotype was seen in
available. The various methods are elegantly presented        5/13 (38.46%) with mean CR1=11.8 months, pseudo
in a review article by Van Deerlin and Leonard (2).           diploid in 6/13 (46.15%), with mean CR1= 6.66
Though the methods of Detection of MC very well               months. The third group was patients having Pre B
established yet its clinical significance is still far from   phenotype, the mean total leukocytic count was 39.83,
it. In certain conditions, the significance is quite well     massive organomegaly was seen in 8/14 (57.14%.)
evident. For instance in MC when the percentage of            normal karyotype was found in 6/14 (42%) with mean
recipient cells is increasing in conesquentive assays,        CR1=12 months, pseudodiploid in 5/14 (35%) with
this for sure would herald the occurrence of relapse.         min CR1=16.4 months, hyperdiploid in 2/14 (14.28%)
Also in cases of pancytopenia after allogeneic BMT            with mean CR1=24.5 months and hypodiploidy in only
detection of chimerism will discriminate failure of           1/14 (7.14%) with mean CR1= 6 months. The last

                                                                                                          9. Leukemia

group was patients with T cell surface markers 7/40           patients and may be due to small number of patients or
(17.5%), their mean age was 8.14 years, with M/F of           we can conclude that immunophenotyping can be used
6:1, the mean total leucocytic count was 33.1, massive        only on the purpose of correct diagnosis not as a
organomegaly in 85.71%, pseudodiploid in 5/7                  prognostic factors. P53, Bcl-2, MDR as a prognostic
(71.42%) with mean CR1= 9.4 months, hyper diploidy            factors in our work reveal that p53 is a good prognostic
in 1/7 (14.28%) with mean CRl=14 months. Only one             indicator of survival in AML patients when not
patient was biphenotypic, a female ten years old who          expressed, MDR when highly expressed in ALL
had massive organomegaly and a total leukocytic count         patients have poor prognosis on treatment outcome and
171.000, the mean CR1 was 4 months and she had both           survival of that patients and Bc12 is of clinical
bone marrow and central nervous system relapse.               significance when highly expressed in both AML and
                                                              ALL patients have a bad prognosis on both treatment
9.32. A Study of Prognostic Factors in                        outcome and survival. Cytogenetic study can be used
Adulthood Acute Leukemias Revealed By                         as a prognostic marker, our result reveal that patients
Surface Markers & DNA Analysis                                with abnormal karyotyping was resistant to treatment
Maha Ibrahim Ismael Ibrahim (MD Medical Oncology,             in both AML and ALL patients.
Mostafa Mahmoud El-Serafi, Sameh Sayed Ahmed                  9.33. Cell Therapy: Utilization of Unrelated
Shamaa, Farida Hassan Abdel Rahman.                           Allogeneic     Hematopoietic      Stem    Cell
                                                              Transplantation     in   Pediatric   oncology
The aim of the present work is to study some                  Hematology
prognostic factors revealed to have impacts on                Mohamed Sedky Mahmoud Sedky (MD Ped Oncology
treatment outcome and survival of adult acute                 2002)
leukemias patients (AML and ALL). Forty five adult            Eliane Glackman, Hossam Kamel Mahmoud, Nagwa
patients with acute leukemia, thirty patients AML and         Abdallah Mahmoud
fifteen patients with ALL were included in the study.
The age ranged from 19-60 years for patients with             Hematopoietic stem cell transplantation is actually
AML and between 16-60 years for patients with ALL.            considered as the first choice effective therapeutic
The male to female ratio was 1:1.3 for AML and 2:1            modality for a variety of malignant and non-malignant
for ALL. Bleeding and pallor were the main presenting         hematological disorders in pediatrics. In spite of the
symptoms in AML while fever and pallor were the               continuous expansion of the transplantation indications
main symptoms in ALL. A higher incidence of                   from one year to the other, the availability of a suitable
lymphadenopathy, splenomegaly and hepatomegaly                donor remains the key of success of such a procedure.
were detected in ALL compared to AML cases. The               Confronted with a limited HLA identical sibling donor
CR rate in AML patients in the present study was              to 30% of cases worldwide, our target is to favor other
33.3% and 53.3% for ALL patients. So, a more                  acceptable hematopoietic stem ce11 sources. Along
intensification of therapy may be needed. The median          more than one decade, a group of 82 children with
follow up of our patients was seven months (210 days).        different malignant or non-malignant hematological
50% of AML patients and 33% of ALL patients died              disorders, and lacking all identical sibling donor, were
during the study, 20%, 1.3% of AML and ALL patients           the subject of allogeneic unrelated hematopoietic stem
still alive diseased at the end of the study, this denoting   cell transplantation in Saint-Louis hospital, Paris. So
that many different prognostic factors and not only the       marrow sources were taken at a first choice in 48
therapy play a significant role on the treatment              patients from a phenotypically identical unrelated
outcome and survival of acute leukemias patients.             donor through the bone marrow donor registries
Some of these prognostic factors were studied in this         starting from end 1987 (NTCDM group), or if
work on the clinical level e.g. age, sex, organomegaly,       unavailable, an unrelated cord blood till to 2
on the laboratory level e.g. Hb level. TLC, platelets         antigen-mismatch from cord blood banks, in 11
number and percent of blast in the peripheral blood, on       patients starting from mid 1994 (UCB group). In a
the immunophenotyping study of surface markers level          determined period between mid 1995 and mid 1999
e.g. CD13, CD33, CD34 in AML and CD2, CD7, CD                 and in order to control lethal GVHD in high-risk
19 in ALL and on molecular biological study by                patients for transplant related mortality, we started all
flowcytometry of p53, Bc12, MDR 1-Pgp, DNA ploidy             experimental phase I-II study in 23 patients, using tile
and lastly cytogenetic study of abnormal chromosomal          positive selection of CD34+ cells as a stem cell source
structure and number. Neither age, sex, Hb level,             (TCDM group). A High-risk GVHD, moderate to
WBCs count nor platelets count were related to the            high-risk infections and rejection characterized the
success of obtaining complete remission in both AML,          NTCDM group arid thus a high transplant
ALL patients and the only detected significant                related-mortality, but as expected there was a low risk
parameter is the blast cell percentage in AML patients        relapse. The survival was low reaching around 25%.
where patients with no response to treatment have a           These results were notably ameliorated by the TCDM
peripheral blast percent of 69.7% and that patients in        by positive selection of CD34+ cells with a remarkable
complete remission have a 49.6% blast percent. When           lower incidence of GVHD, thus a act lower TRM.
we use surface markers as a prognostic factor in our          Although a higher rate of relapse, moderate to high
study no significant relation was found between surface       incidence of infections an rejection occurred with a
markers and treatment outcome in both AML, ALL                doubtful beneficial effect of the negative fraction add

Scientific Annual Report 2000-2002

back in these cases. TCDM showed the best survival          significantly increase of GRX in most of hematological
rate around 45%. The UCB group was characterized by         malignancies. There was a positive correlation between
a high TRM either due to infections, toxicity or            GPX and SOD in most of the studied groups. In
GVHD. Unexpectedly, this group showed a high                contrast, breast cancer patients showed significantly
probability of engraftment. GVHD with no case of            increase of the activity of SOD, GPX and GRX when
relapse. So the result was a low survival around 10%.       compared to the control group. On the other hand,
                                                            urinary bladder cancer patients showed only significant
9.34. Acquired Von Willebrand Disease (Avwd)                decrease of GPX activity in relation to the control
in Haematologic Malignancies                                subjects.
Walid Monir Ahmed (MD Clinical Pathology).
Magda Mahmoud Ahmed Assem, Naguib Zoheir                    9.36.         Disseminated       intravascular
Mostafa, Maha Saleh                                         Coagulopathy and Its Relation To Phenotype
                                                            in Acute Leukemia
Acquired von Willebrand disease is an acquired              Manar Mohamed Mohamed Ismail (MD Clinical
bleeding disorder which may suddenly become                 Pathology, 2001)
manifest in individuals, in the absence of a                Moshira Fathy El-Seify, Hossam Mohamed Kamel,
past-personal history of bleeding and frequently in         Lobna Abdel-Azim Refaat.
association      with     monoclonal      gammopathies,
lymphoproliferative,        myeloproliferative        and   Hemorrhagic diathesis is the commonest cause of
autoimmune disorders. 114 patients attending to the         morbidity and mortality in acute leukemia. It is most
Medical Oncology department, NCI presenting with a          commonly due to thrombocytopenia associated with
recent     development      of    non-thrombocytopenic      the proliferating leukemic clone and with cytostatic
mucoutaneous bleeding were included in this study. No       treatment given. However, in a significant number of
single test is sufficiently enough to diagnose AvWD         cases, disseminated intravascular coagulation (DIC)
but the diagnosis requires multi-test analysis. Screening   plays an important role. Previously, it was thought that
tests were performed in addition to specific tests. Using   DIC was mainly confined to acute promyelocytic
ROC analysis, Ri:Cof was found to be the best               leukemia, but recently it has also been reported to
diagnostic test. AvWD was most frequent in CML              occur in other subtypes of acute leukemia. The
patients and was prominent in patients with blood           pathogenesis and mechanism of tills disorder is a
group O. Type 2.A with loss of HMWM was most                complex one. The most important of these relates to the
frequent in AvWD. Conclusion No single test is              leukemia, cells circulating in the blood which may
sufficiently enough to diagnose AvWD but a                  contain procoagulants, fibrinolytic activators or
combination of screening and specific tests may             proteases. These substances can be released into the
increase the total accuracy of the diseases diagnosed.      blood from disisntegrating cells during cytostatic
One ratio Ri Cof/vWF:Ag may be used to predict type         treatment. Coagulation disorder may also occur due to
2M and allow better identification of the different         leukemia - associated complications such as sepsis or
subtypes of AvWD. CIVIL is the most frequent                Hepatic impairment. The aim of the present work was
hematological disorder affected by AvWD followed by         to find out the frequency of DIC and pre- DIC in newly
NHL, CLL and MM. Blood group O is most frequent             diagnosed acute leukemia cases both at presentation
blood group affected by AvWD while group AB is the          and in the first seven days of remission induction and
least affected. The most common subtype of AvWD is          to attempt to correlate such findings with
type 2A (with loss of HMWM).                                immunophenotyping. The work was carried out on 62
                                                            newly diagnosed acute leukemia cases, 31 ALL, and 31
9.35. A Study on The Antioxidants in                        AML. Both groups were subjected to the following:
Leukaemias and Solid Malignant Tumors                       1- Complete blood picture 2- Bone marrow
Tarek Mahmoud Ali El Masry (MD Clinical Pathology           examination 3- Cytochemistry 4- Immunoperoxidase
2001)                                                       staining for immunophenotyping 5- PT, PTT,
Salwa Abdel Hamid El-Demerdash, Mohamed Ibrahim             fibrinogen level and FDP test as laboratory evidence of
El Khodary, Khaled Mostafa Aboul Enein                      DIC 6- D-diner and TAT tests as laboratory evidence
                                                            of pre-DIC. Groups and 6 were done at presentation as
All cells have an antioxidant defense systems that          well as on the second and seventh days of remission
consist of interacting components. The most important       induction. Prolongation         of   PT, PTT          and
are SOD, glutathione and its related enzymes and CAT.       hypofibrinogenemia denoted activation of coagulation
The balance between these several antioxidant enzymes       and thrombin generation, while the presence of FDPs
rather than the activity of a single component may          and D-dimer denoted plasmin generation and
determine the degree of protection. Our study included      secondary fibrinolysis. At presentation DIC was
78 patients of different ages and sexes with different      diagnosed in 38.7% of ALL and 61.3% of AML cases,
hematological malignancies, 16 male patients with           on the second day in 58% of ALL and 80% of AML
urinary bladder cancer and 15 female patients with          and on the seventh day postinduction it was diagnosed
breast cancer. They were all studied as regard the          in 34.6 % of ALL and 66.6% of AML cases, being
activity of erythrocyte GPX, SOD and GRX and were           statistically significant and denoting higher incidence
matched with control healthy subjects. Both GPX and         in AML in all cases. The occurrence of DIC had no
SOD were found to be significantly decreased with           significant correlation with phenotypic profiles of acute

                                                                                                         9. Leukemia

leukemia, age, sex, clinical signs or hematological           In the current study, cytogenetic analysis was done for
findings. The results reported here indicate that there is   33 patients suffering from ALL, 15 suffering from
a chronic activation of blood coagulation in most of the     AML, and 15 suffering from CML, by both
AL patients. DIC can develop in various forms and            conventional cytogenetic analysis (G-banding) and by
subtypes of acute leukemia, mostly promyelocytic and         FISH using LSI probe for t(12;21), t(9:22) and WPC of
monocytic leukemias both at the onset of the disease         chromosomes 1 and 19 (or t(1;19) for some selected
and after cytostatic treatment. Because of these             ALL cases, LSI probe for t(15;17) for some selected
findings, we suggest that an analysis of the DIC             AML cases in LSI probe for t(9;22) for detection of
syndrome should be made at diagnosis and during              Philadelphia chromosome in CML cases. Successful
remission induction in all acute leukemia patients. It is    Karyotyping was encountered in 25/33 (75.8%) in ALL
essential to predict the probability of DIC (pre-DIC) by     patients, 12/15 (80%) in AML patients and 9/15 (60%)
determination of the molecular hemostatic marker             in CML patients. Both numerical and structural
(D-dimer and TAT) for early diagnosis and treatment          chromosomal aberrations were detected in l3/25 (52%)
of this complication to have an effective treatment          of ALL cases, 4/12 (33.3%) of AML cases and 5/9
outcome.                                                     (55.5%) of CML cases were positive for Philadelphia
                                                             chromosome. Using FISH analysis with LSI t(12;21)
9.37. Deoxyribonucleic Acid Ploidy, Nucleolar                for ALL patients revealed positive results in 7/20
organizer Regions and Morphometric Analysis                  (35%), while with WPC of both chromosomes 1 and 19
By Image Cytometry IN Pediatric Acute                        for t(l:19) for the same ALL patients revealed positive
Leukemias                                                    results in 3/20 (15%). Philadelphia chromosome were
Somaya Mohammed Abdel-Moneim Ramadan (MD                     positive in 2/6 (33.3%) selected ALL patients. Using
Clinical Pathology 2001)                                     FISH analysis with LSI t(15;17) for AML patients
Mostafa I. Aboul-Enein, Somaya A. K. El-Hosseiny,            revealed positive results in 5/9 (55.5%), while LSI
Sherif A. Aboul-Naga                                         t(9;22) for CML patients revealed positive Ph
                                                             chromosome in 11/15 (73.3%). Conventional
The aim of this work is to study the morphornetric           cytogenetic study was found to be less successful in
features of the malignant leukemic cells, nuclear DNA        leukemic cases where low number of metaphases and
ploidy and proliferation rate as useful parameters in        poor duality of mitosis is usually obtained. On the
predicting the biologic behavior of ALs in children.         other hand, FISH provides a simple, rapid and
This study involved 45 subjects (35 pediatric acute          unequivocal cytogenetic diagnosis.
leukemia patients who had received no prior therapy
and 10 healthy children as a control group of matching       9.39. Salvage Therapy Protocol For Refractory
age and sex. These patients were selected from the           or Relapsing Acute Lymphoblastic Leukemia
clinical pathology and pediatric oncology departments,       and Disseminated Lymphoma in Children
NCI, Cairo University. Of these patients it was possible     Shimaa Lotfy Abo-El Gouth Nassif (MS Pediatrics,
to follow 34 cases for 2 years. These cases were             2002)
subjected to thorough history taking, clinical               Aly Mostafa Aly, Sherif Aboul Naga, Iman Abd
examination and laboratory investigations (complete          El-Mokhales Sidhom
blood picture, bone marrow aspiration, cytochemical
staining). DNA ploidy, nucleolar organizer regions and       Acute leukemia is the most common cancer in children,
morphometric analysis were carried out by CAS 200            with acute lymphoblastic leukemia (ALL) the most
image analyzer of bone marrow (BM) and peripheral            common subtype. Despite improvements in the
blood (PB) samples once at diagnosis of ALs. Results         treatment of pediatric acute lymphoblastic leukemia,
revealed the following. 1. The incidence of aneuploidy       approximately one in five patients will develop
in AL (31.4%). 2. No relation between DNA ploidy             recurrent disease. The ultimate objective of the study
pattern and remission rates of ALs. 3. Ag-NOR                were to reinduce and maintain disease remission for
enumeration using silver staining technique may be           children with relapsed ALL using the subjected
significant in the evaluation of pediatric AL.               protocol of salvage therapy and to evaluate its impact
Measurement of nuclear area in AL cells helps to             on prolonging EFS and RFS. Also the study was
characterize L1 and L2 subtypes of ALL and to predict        designed to identify patients in relapse or those who are
the prognosis of acute leukemias. Automatic image            resistant to therapy, analysis of underlying risk criteria
analyzer offer the advantages to detect small aneuploid      and determining the response rate and toxicity to
population and proved to be the most recent approach         mufti-agent combination therapy used in different
for standard Ag-NOR and morphometric analysis.               phases of therapy. This study was carried out on 51
                                                             pediatric patients with definitive diagnosis of refractory
9.38. in Situ Hybridization As A Tool For                    or relapsing acute lymphoblastic leukemia in first
Detection of Chromosomal Anomalies in                        relapse. Of the fifty-one patients who were included in
Hematopoietic Malignancies                                   the study, two were refractory (4%) and 49 (96%) with
Omyma Mohamed Ahmed Hassanin (MD Clinical                    acute lymphoblastic leukemia in first relapse. Isolated
Pathology, 2001)                                             medullary relapse was present in 39/49 patients
Azza Mohamed Kamel, Wagida Abdel Rahman                      (79.6%), isolated extramedullary relapse in 5/49
Anwar, Sherif Abo El Naga                                    patients (10.2%) [Four with isolated CNS 8.2% and
                                                             one with isolated testicular 2%] and 5/49 patients

Scientific Annual Report 2000-2002

(10.2%) with combined relapse [two combined               5q 13 translocations and deletions FISH was performed
hematological and CNS (4.1%) and three combined           with probe PAC 125D9 to delineate the breakpoint on
hematological and testicular (6.1%)]. By the end of 31    chromosome 5q 13 and results were as follows: the
November 2000, 26 patients (51%) were alive in            breakpoint was telomeric to the probe in group A while
second complete remission, 22 patients (43.1%) off        centromeric in group B. In group A, 13/22 cases
therapy (three resistant and 19 relapsing) and three      showed snore signals oil the del5q 13 than the normal
patients (5.9%) died (Two during re-induction and one     5q 13. FISH could also be of benefit in detecting new
during intensive module one cycle I). The estimated       chromosomal aberrations such as t(l;19)(q41;?) that is
two- year EFS was 38.5%, while the estimated two          different from the known t(l;19)(q23;p 13). FISH was
year RFS was 42.1%. EFS and RFS in relation to age,       also a tool in detecting an amplification (>10 signals in
sex, hepatomegaly, FAB and IPT showed no statistical      60% of interphase nuclei) of the AML1 gene in an
significant differences. On the other hand the two year   ALL case. To our knowledge the literature is poor in
probability of RFS was 77% for TLC <10 x109 /l            this finding. Ultrastructure features of hematopoietic
versus 28% for TLC > 10 x109 /I. Results are              malignant cells were correlated with their cytogenetic
statistically significant (p = 0.039). Also RFS in        aberrations.       Nucleocytoplasmic        asynchrony,
relation to splenomegaly, duration of 1st CCR and         deformation of nuclei, abnormal large mitocliondria
initial line of therapy shows significant statistical     with destruction of their crestae and very scarce
difference (P value <0.05). Toxicities to different       primary granules in AML-M2/t(8;21) but not in
phases of treatment were recorded; with a higher          AML-M2 with normal karyotype. Also, in
incidence of sever myelosuppresion (neutropenia grade     APL/t(15;17), increased vacuolations, abnormal large
4) following consolidation 1 (FLAG) however               granules, dilated endoplasmic reticulum, and many
mortality related septic episodes were not documented     nuclear split were detected.
in relation to this phase of treatment (FLAG). The
subjected protocol of salvage therapy was tolerable and   9.41. Protocol Therapy For Newly Diagnosed
successful in achieving two year RFS of 42%               Acute Lymphoblastic Leukemia Different Risk
compared to 5 % in previous protocols used in our         Groups
institute.                                                Inas Mohsen El Said El Nadi (MD Ped. Oncology,
9.40. Ultrastructural, Molecular (FISH), and              Mohamed Hani Hussien, Azza Mahmoud Kamel,
Cytogenetic Abnormalities in Blast Cells                  Sherif Abo El Naga.
Sherine lbrahim Salem El-Deghaidy (MD Clinical
Pathology, 2001)                                          This prospective study included 152 newly diagnosed
Mostafa I Aboul-Enein, Amira Khorshed, Farida H           ALL patients below the age of 20 years, who presented
Gadalla,                                                  in the pediatric oncology clinic of the National Cancer
                                                          Institute, during the period between 1st of December
This study was performed on 103 patients with             1998 to 1sT December 2000. Patients were followed
different hematological malignancies. Fifty two           until April 2002. They received one protocol; cranial
patients presented to the NCI, Cairo University and the   radiotherapy was restricted to patients with initial CNS
rest presented to Hôpital 1'Archet 2, Nice, France The    disease or high-risk patients. Instead, CNS prophylaxis
patients were divided into 2 groups: Group 1 included     to other patients was through extended triple intrathecal
cases with different chromosomal aberrations. Group 2     therapy with intermediate or high dose systemic
included cases with chromosomal aberrations of 5q13       chemotherapy. The most common encountered
These cases were subjected to (1) CBC, BM,                immunophenotype was the pre-B, which presented
cytochemistry, and immunophenotyping (on routine          33%, followed by the T phenotype 29.1 %, C-ALL
basis) (2) Karyotyping (3) FISH (4) EM. Group 1           category constituted 26.7% and early Pre-B was 5.5%.
patients include 42 patients with lymphoid                By the end of follow up period (41 months) 57.8%
malignancies, having ALL- L1, ALL-L2, ALL-L3, and         achieved CR, 11.2% developed HR, 5.3% developed
NHL. Clonal chromosomal abnormalities were                CNS relapse and one case developed both HR and CNS
detected in 33 cases. Only 3 cases had normal             R. The OS to the whole group was 83.3%, the
karyotypes. The t(9;22) and hyperdiploidy were            estimated DFS for the whole group was 76.6% and the
recurrent chromosomal anomalies. Four cases showed        estimated EFS were 64.7%.
the t(8;14), one of which had Pre-B cell
immunophenotype and an additional t(1;19). Two cases      9.42. Light and Electron Microscopic Changes
had t(12;21)(p 13;q22) with TEL/AML1 fusion gene.         in Bone Marrow Cell Subjected To Drugs and
Group 1 also includes 35 patients with myeloid            Chemicals: An in Vitro Study
malignancies including AML, CML and MDS. Clonal           Basma Mostafa El Gamal (MD Clinical Pathology
chromosomal abnormalities were detected in 29 cases.      2000 )
Three cases had the t(8;21). Eight cases lead the         Amira Mokhtar Khorshed, Amr Hammam Ziada,
t(15;17). Seven cases had chromosome 16 structural        Khaled Mostafa Aboul-Enein
abnormalities. Group 2 patients with RA, RARS,
RAEB, CMML, AML, NHL was further divided into 2           The human body is at a continuous defense against
groups: The Group A: 22 patients with del5q 13 The        extraneous agents in its environment. This host defense
Group B: 4 patients with complex karyotype including      is directed towards bacteria, viruses, parasites ...etc.

                                                                                                        9. Leukemia

Besides these infectious agents. the body is exposed to     g/dl with a mean 6.7 g/dl. CSF was free of blasts in all
environmental pollution or agents it is forced to use       patients while chest X-ray was positive for
such as chemicals, drugs and irradiation. These             mediastinal/hilar lymphadenopathy in 20% of patients.
damaging      agents     affect    immunity,     cellular   The most commonly encounterd FAB subtype was M1
development. Chromosomes, DNA, RNA, enzymes,                in 34.9% followed by M2 in 33.9% and M3 in 16.5%.
proteins and other cellular components. In response to      6.3% of our patients were diagnosed by
the changes, which develop, the body could either           immunophenotyping after being misdiagnosed by BM
repair the damage in DNA or others or the process           aspirate. A wide panel of immunophenotyping was
progresses to death of the cells. Damage is elicited into   done including MPO, CD13, CD33, CD14 and CD15.
several morphologic changes in the form of necrosis or      The most commonly detected IPT was MPO in 98% of
enhancement of the normal programmed cell death or          cases followed by CD13 (86%), CD 33 (72.4%) and
apoptosis. Such a stimulus may be in the form of            HLA-DR (60.4%). Correlation with survival was done
chemotherapeutic       agents.      Apoptosis      shows    with no statistical significance. Eighty-one patients
characteristic morphologic changes that are quite           received induction chemotherapy with a CR ratio 85%,
distinct from those seen in necrosis. These changes         while 15% of patients failed to achieve remission. 32
could be detected microscopically, either by electron or    patients received chemotherapy on Arm A with
by light microscopy. This study involved the                consolidation and maintenance chemotherapy for total
exploration of the toxic damage to normal and               duration of 18 months. Treatment related deaths were
leukemia bone marrow cells by some of the                   low (6.4%) while relapse rate was high (40%). OS
chemotherapeutic agents in common use. Bone marrow          ranged between 4-30 months with a mean of 13.2
cells were subjected in vitro to these agents, then         months. DFS for the same group ranged between 3-29
studied by light and electron microscopy. Aspirated         months with a mean of 12.1 months. Thirty-one
bone marrow of 10 acute myeloid leukemic patients, 10       patients received chemotherapy on Arm B with
acute lymphatic leukemic patients and 10 non                consolidation therapy without maintenance for duration
malignant cases wets incubated in culture medium at         of 6-8 months. Treatment related deaths (22.3%), while
37°C for 24 hours. Before incubation, selected              relapses were 13%. OS for the second group of patients
chemotherapeutic agents were added to culture tubes at      ranged between 3-25 months with a mean of 12.1
calculated dose levels commonly used in treatment.          months while DFS for the same group ranged between
After 24 hours incubation, Bone marrow examination          2-23 months with a mean of 10.8 months No statistical
was done by light microscopy and compared to its state      significance was found between both groups.
before incubation. Electron microscopic examination
was done for some selected cases showing, prominent         9.44. Study of Some Prognostic Factors in
necrotic or apoptotic changes. The sensitivity of the       Chronic Lymphocytic Leukemia
different cell lines to chemotherapy was estimated.         Mohamad Ahmad Mohamad Al-Akkad (Internal
Also, the apoptogenic effects of the different              Medicine)
chemotherapeutic agents on individual bone marrow           Zekri Khaled Zekri, Ibrahim Amin Ibrahim, Azza
samples was compared.                                       Mahmoud Kamel, Hussein Khaled

9.43. Management of Childhood Acute                         B-cell CLL is characterized by a highly variable
Nonlymphocytic Leukemia at The NCI The                      clinical course. At present clinical stage remains the
Impact of An integrated Approach on Survival                strongest predictor of survival in patients with B-CLL.
Hany Abdel Rahman Sayed (MSc Pediatric Oncology)            The substantial heterogeneity even within clinical
Salah Saleh Abd-El Hadi, Heba Mohammed Hamed                stages has led to a search for more relevant prognostic
Shaker, Lobna Mohammed El-Amin Shalaby                      factors. The purpose of this study was to investigate the
                                                            clinical significance of the expression of the
This study was conducted in the Pediatric Unit of the       proliferative markers PCNA and Ki67+ cells as well as
Medical Oncology Department, National Cancer                the oncoproteins bcl-2 and p53 in a series of CLL
Institute-Cairo University during the period from           patients. The study was carried out on 48 cases. These
October 1996 till March 1999. It included 113 patients      included 28 males and 20 females with an age range of
≤16 years of age newly diagnosed as AML. The                35-80 years. All cases presented to the outpatient
Patients ages ranged from 1 - 16 years with a mean of       clinics of the Medical Oncology Department of the
8.2 years. Male to Female ratio was 1:1. Most of our        NCI, Cairo University. All cases were subjected to the
patients were from the area of greater Cairo (45%),         following      investigations:    complete      physical
other patients come from El-Menya, Fayoum, Kena             examination, complete blood count, BM examination,
and Bany Sweif. Pallor was the most common feature          and immunophenotying using MoAb and the
at presentation (95%) followed by fever (51.3%) and         immunoperoxidase staining technique on the
bleeding tendency (41.6%). Hepatomegaly was                 mononuclear cytopreps. According to the modified Rai
detected in 17.7%, while splenomegaly was seen in           staging system, only one patient was in stage 1,
35.5%. Lymph node enlargement was seen in 23.9%,            32(68.1%) in stage 2, and 14(29.8%) in stage 3. Forty
while granulocytic sarcoma was detected in 10.6% of         cases (93.00%) belonged to B-CLL while only 3 to
our patients. The initial total leucocytic count ranged     T-CLL. CD 10 positivity was observed in 36 cases
from 0.1/ul to 386.000/ul with a mean of 48.200/ul          (78.30%). Bcl-2 expression was detected in 13 cases
while hemoglobin level ranged from 1.0 g/dl to 12.2         (31%) and p53 in 7 cases (16.70%). Positive

Scientific Annual Report 2000-2002

expression of proliferative markers Ki-67+ cells and        failure in patients with hematopoietic malignancies and
PCNA were observed in 7 (17.10%) and 24 (60%)               suffer from cytopenias. To reach this goal bone marrow
cases respectively. There was no difference with regard     trephine biopsies were applied to 120 patients
to the distribution of the studied biologic markers         complaining from variable degrees of anemia,
among the different disease stages except for Ki-67         bicytopenia, or pancytopenia either at their initial
positivity which showed a significantly higher              presentation with hemopoietic malignancies or during
incidence in stage 3 disease (p=0.017). The                 the course of their therapy. These patients included 90
proliferative activity was not different between p53 +      male and 30 female with M: F ratio of 3: 1. Their ages
and p53- groups, while bcl-2 expression was associated      ranged from 2 to 73 yrs. for males and from 3 to 66 yrs.
with a significantly reduced proliferative activity         for females. All these patients were suffering from
(p=0.042). Among 13 patients who expressed the bcl-2        hematopoietic malignancies characterized either by
protein 5 patients coexpressed p53. The L1 of bcl-2         focal BSI lesions or by alterations of the BM
was not different between the two groups indicating         architectures, e.g., AA, MDS, MF/MS, CML, CLL,
that p53 has no impact on the expression of bcl-2. The      HCL, HD and NHL. They were subjected to CBC, BW
finding drat bcl-2 is associated with a low proliferative   aspiration and BM biopsies. The patients with HD and
activity is consistent with recent in vitro data showing    NHL were subjected to bilateral biopsies, while the
that the bcl-2 protein retards entry into cell cycle. Out   other groups were subjected to unilateral satisfactory
of 45 evaluated patients, six patients (13.3%)              long care biopsy. In the late: groups, bilateral biopsies
experienced complete response, 30(66.7%) showed             were done only whenever required. All the obtained
partial response, and 9 (20.00%) showed no response.        biopsy sections were subjected to HOE staining as well
Only the modified Rai staging and p53 expression            as a panel of different special stains including iron
among all the studied disease parameters showed a           stain, reticulin stain, giemsa stain and toluidine blue
relationship to treatment outcome. Out of 31 patients       stain. According to requirements, some cases were
with stage l and 2 disease, 28 patients (90.32%) bowed      subjected to IHC staining using different monoclonal
a response and only 3 showed no response. On the            antibodies. Conclusion: BM biopsy is considered an
other hand, only 8 out of 14 patients who had stage 3       indispensable diagnostic tool in different hematopoietic
disease showed an objective response (p=0.019). Of the      malignancies that presenting with variable degrees of
six patients showing positive p53 immunostaining,           cytopenias. Application of a panel of special stains as
only 3 patients showed a response while 30 out of 34        well as immunohistochemical staining on the biopsy
patients negative for p53 staining showed a response        sections can significantly help the assessment of
(p=0.05). Of the 47 patients followed for at least 3        pathogenesis of bone marrow failure in these
years, the percentages of overall survival after one, two   hematopoietic malignancies.
and three years of follow up were 97.9, 89.4, and 80.3
respectively. Both bcl-2 and p53 expression affected        9.46. The Heterotopic interaction Between The
patients' survival. The percentages of survival of          B1-integrin VLA-4 and Its Ligand The Vascular
patients showing positive immunostaining for bcl-2          Cell Adhesion Molecule (VCAM-1) in The
after 1, 2, and 3 years were 92.3, 76,9 and 53.9            Various Hematological Leukaemia
respectively, while these percentages for patients          Mona Abbas Abdel Sattar (MD Clinical Pathology &
showing negativity for bcl-2 staining were 100.0, 93.1      Oncologic Laboratory Medicine 2000)
and 89.4 (p=0.039). As regards the expression of p53        Moshira Fathi El Saify, Fatma Mohamed Nasrat,
oncoprtein, the percentages of survival at 1, 2, and 3      Rabab Mohamed Gaafar, Sherif Naseh Amin
years for patients with positive staining were 100, 71.4,
and 42.9 respectively. On the other hand, these             This work included 152 patients comprising 66 ALL,
percentages were 97.1, 91.4, and 85.4 for patients with     52 AML and 34 CLL. Each group was analysed for the
negative staining (p= 0.024). In conclusion, our            expression of certain adhesion molecules. Expression
findings indicate that bcl-2 and p53 expression CLL is      of VLA-4 (CD49d) as well as the expression of its
associated with short survival. In the context of a         ligand VCAM-1 (CD 106) has been studied in all the
heterogeneous condition like CLL, the simple                three    groups.     Additionally,     expression     of
inexpensive and reliable ICC method appears capable         NK-associated markers (CD56 and CD 16) as well as
of identifying patients who may be considered               CD 11b along with DNA ploidy and cell-cycle analysis
candidates for more intensive therapeutic strategies.       for 5-phase cells and the proliferative index were
                                                            carried out in the AML group. Each of the above
9.45. The Diagnostic Role of Bilateral Bone                 parameters was correlated to age and sex; clinical
Marrow Trephine Biopsy with New Staining                    features, laboratory features at presentation, FAB
Methods in The Assessment of The                            subtype, immunologic subtype and response to
Pathogenesis of Bone Marrow Failure During                  treatment. Results revealed the following: In the ALL
The Course of Hematopoietic Malignancy                      group, VLA-4 expression was not seen to be related to
Amr El-Sayed Zaher (MD Clinical Pathology 2000)             any of the parameters studied. The fact that it is
Amira Mokhtar Khorshed, Nadia Mahmod Mokhtar,               expressed on normal hematopoietic cells makes it a
Khalid Mostafa Aboul-Enein                                  useless diagnostic tool and the fact that its expression
                                                            did not affect treatment outcome makes it an
This study is to evaluate how the BM trephines biopsy       inadequate prognostic tool. As regards VCAM-1
with different stains disclosing the pathogenesis of BM     expression, it was not seen to be related to age, sex,

                                                                                                       9. Leukemia

distinct clinical, laboratory features or immunologic       This decrease was significant in M2, and in M3. And it
subtypes. However, it was shown to he significantly         was non-significant in M0-1, and in M4-5. In AML,
related to treatment outcome with a worse complete          and ALL samples, zinc showed positive correlation
remission rate among those expressing the marker thus       with copper. 3. Copper: (a) In ALL: Copper showed no
possibly having prognostic implications in the patient      significant differences between ALL group and the
populations studied. In the CLL group, neither VLA-4        control group. (b) In AML: Copper showed
nor VCAM-1 related to sex or clinical features. On the      significance decrease in AML cases as compared to the
other hand, they both related significantly with platelet   control. This significant decrease was only observed in
count and clinical staging where higher marker              M3 subgroup. As regards AML group, copper showed
expression was seen with platelet counts ≤ 100x109/L        a high positive correlation with Cu/Zn ratio. 4.
and in stages III and IV. As regards response to            Copper/Zinc ratio: (a) In ALL: This ratio was
treatment, VLA-4 pressing cases were seen to respond        significantly higher in L1 and non-significantly higher
less than VLA-4 negative ones. In the AML group,            in L1, and in L2, as compared to the normal control.
VLA-4 expression did nut relate to any of the               (b) In AML: This ratio showed non-significant increase
parameters studied making it a useless tool in both         in M0-1, M2, M3, and in M4-5 as compared to the
diagnosis and prognosis. As regards VCAM-1, CD11b           control. In AML group, this ratio was positively
and CD56 expression, all were seen to be I related to       correlated to LDH, while it showed a negative
the monocytic subtypes of the FAB classification.           correlation with zinc and a higher positive correlation
Neither VCAM-1, CD16, CD56 nor CD11b related to             with copper. In ALL, it also showed negative
age, sex, clinical and laboratory features or response to   correlation with zinc and high positive correlation with
treatment. DNA, ploidy also did not relate to age, sex,     copper. The statistical comparative analysis in between
laboratory features, marker expression, or response to      ALL, and AML, and between the different groups of
treatment. However, diploid cases had significantly         ALL and AML studied did not show significant
higher WBC count at presentation and bone marrow            differences as regards trace elements and biochemical
blast cell percentage. Proliferation fraction not           markers. Although the role of selenium, zinc, and
correlated to any of the parameters.                        copper in the carcinogenic process is being thoroughly
                                                            investigated, and has proven to be of value, further
9.47. Selenium and Other Trace Elements in                  studies are recommended to delineate their roles in
Acute Leukemia                                              initiation versus nepotistic progression of tumors.
Niveen Abdel Gawad Ahmed Omran (MD Clinical
Pathology)                                                  9.48. Acquired Thrombotic Disorders in Acute
Niveen M. El Hefawy, Lobna A. Refaat, Naglaa M.             Leukaemia and Non Hodgkin's Lymphoma
K.Ismail                                                    Nahla Ahmed Bahgat Abd El-Latif (MD Clinical
                                                            Pathology and Oncologic Laboratory Medicine 2000)
This work was intended to study the changes in some         Amira M. Khorshed, Magda M. Assem, Amr H. Ziada,
serum trace elements, and some biochemical markers
in acute leukemia. This study involved 80 cases with        The aim of the present work was to identity
acute leukemia classified according to FAB                  abnormalities in haemostasis that relate to thrombotic
classification as follows: ALL group: 40 cases, of          or haemorrhagic manifestation in patients with acute
which 12 cases were L1, 23 cases were L2, and 5 cases       leukaemia and NHL. The work was carried out on fifty
were L3. AML group: 40cases, of which one case was          patients 30 with acute leukaemia and 20 with NHL.
M0, 11 cases were M1, 15 cases were M2, 3 cases             Then patients with ALL two were L1 and eight L2. 20
were M3, 8 cases were M4 and 2 cases were M5. Also,         patients with ANLL seven (M1), seven (M2), one
20 healthy normal age-matched individuals were              (M3), one (M4), two (M5a), one (M5b) and one (M7),
collected as control group. All the cases and the           20 patients with NHL. Ten healthy subjects were taken
controls were subjected to the following: 1. -Chemical      as control group. Different laboratory investigations
analysis for (Calcium, some liver function tests. some      were done including: (1) Routine hematological study
kidney function tests, total lactate dehydrogenase          (complete blood picture, Bone marrow examination (2)
enzyme, uric acid, and cholesterol 2. -Estimation of        screening tests for coagulation (PT, PTT, TT) (3)
some trace elements, namely selenium, zinc and              Specific tests for coagulation (protein C and protein S)
copper. The following, results were observed: 1.            (4)Tests for pathological anti-coagulants (lupus-like
Selenium: (a) In ALL: It was significantly lower in         anticoagulant activity and antiphospholipid antibodies).
ALL groups than in control group. This significant          Results revealed the following: In patients with acute
decrease was observed in all ALL subgroups L1, L2           leukemia and NHL. a reduction in the hemoglobin
and in L3 (b) In AML: Selenium also showed                  levels was found. Platelet count was reduced inn acute
significant decrease in AML group as compared to the        leukemia patients, while in NHL no difference was
control group. This decrease was significant in all         detected as compared to control. Screening coagulation
AML subgroups M0-1, M2, M3 and in M4-5. 2. Zinc:            test; showed a significant prolongation of the (PT,
(a) In ALL: It was significantly decreased in ALL           PTT, TT). Except for TT, which was not prolonged in
samples, than in controls. This significance decrease       NHL. No difference detected between ALL and ANLL
was observed in all ALL subgroups. L1, L-2, and in          protein C and S functional activity was reduced in our
L-3. (b) In AML: Zinc also showed significant               patients compared to normal controls. In ANLL protein
decrease in AML group as compared to the control.           C activity was more affected than ALL. No difference

Scientific Annual Report 2000-2002

was detected concerning protein S between ALL and
ANLL patients. Two tests were done as regards anti-
phospholipids antibodies. First, the lupus anticoagulant
clotting test, prolonged APTT sensitive for the
detection of lupus anticoagulant was found in the
present study High titres of IgG APA was also present
in acute leukemia and NHL patients.


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