Η αναιμία σχετιζόμενη με τη νόσο στη χρόνια λεμφογενή λευχαιμία (ΧΛΛ) είναι διάγνωση εξ΄ αποκλεισμού και διαπιστώνεται στις περιπτώσεις που δεν ανευρίσκεται άλλο εμφανές αίτιο αναιμίας. Δεδομένου ότι η παθογένεια της αναιμίας αυτής δεν είναι διευκρινισμένη, μελετήσαμε διάφορες παραμέτρους της ερυθροποίησης σε πρωτοδιαγνωσθέντες ασθενείς με ΧΛΛ και αναιμία σχετιζόμενη με τη νόσο και τις συγκρίναμε με τα αντίστοιχα ευρήματα ασθενών με ΧΛΛ χωρίς αναιμία και χωρίς να έχουν λάβει θεραπεία και με αυτά των υγιών μαρτύρων. Η διήθηση του μυελού των οστών από τα κακοήθη Β-λεμφοκύτταρα δεν ήταν αποκλειστικά υπεύθυνη για την πρόκληση αναιμίας. Τα CD34+ κύτταρα του μυελού των οστών στη ΧΛΛ δεν εμφάνισαν ενδογενή διαταραχή στη στροφή και περαιτέρω διαφοροποίησή τους προς την ερυθρά σειρά. Επιπλέον, δε διαπιστώθηκαν ανεπαρκή επίπεδα ερυθροποιητίνης ορού ούτε ελαττωματική απάντηση των ερυθροποιητικών προδρομικών κυττάρων στην ΕΡΟ στον άξονα ΕΡΟ-ΕΡΟ υποδοχέα κατά τη διέγερσή τους με ΕΡΟ ± TNF-α. Από την άλλη μεριά, τα επίπεδα του TNF-α βρέθηκαν αυξημένα στους ασθενείς με ΧΛΛ και αναιμία σχετιζόμενη με τη νόσο και φάνηκε ο TNF-α να ασκεί άμεση κατασταλτική δράση στη διαφοροποίηση των CD34+ κυττάρων προς κύτταρα της ερυθράς σειράς in vitro. Η μελέτη μας έδειξε ότι η αναιμία σχετιζόμενη με τη νόσο στη ΧΛΛ δεν οφείλεται σε ενδογενείς διαταραχές των ερυθροποιητικών προδρομικών κυττάρων, αλλά πιθανότατα να αποδίδεται στην απευθείας κατασταλτική επίδραση του TNF-α . / Disease-related anemia in chronic lymphocytic leukemia (CLL) occurs when the obvious causes are excluded while its pathogenesis is still obscure. To investigate its underlying mechanisms we studied parameters of erythropoiesis at cellular and molecular level in newly diagnosed CLL patients with disease-related anemia in comparison with those of non-anemic CLL patients and normal controls. Bone marrow (BM) infiltration by leukemic B cells was not exclusively responsible for the presence and the severity of disease-related anemia and BM CD34+ cells were intrinsically capable of generating erythroid precursors. No deficiency of serum erythropoietin (EPO) or defective intracellular response of erythroid precursors to EPO ± Tumor Necrosis Factor-α (TNF-α) stimulation was also observed. On the other hand, serum TNF-α levels were found increased in patients with CLL and disease-related anemia and TNF-α appeared to exert a direct inhibitory effect on the differentiation of CD34+ cells towards the erythroid lineage in vitro. Our study showed that disease-related anemia in CLL was not due to intrinsic defects of erythroid precursors, but might result from the direct suppressive effect of TNF-α on the erythroid production.

Monoclonal B lymphocytosis (MBL) has been recognized as a novel diagnostic condition. This study aims at the identification of clonal lymphocytosis in the patients with asymptomatic lymphocytosis. A total of 203 patients were evaluated for clonal B and T lymphocytosis by using flow cytometry and multiplex-PCR. Among them clonal B- or T-cells were detected in 54.2% of the cases, of which 38.4% were clonal B-cells and 15.8% were clonal T-cells cases. By immunophenotype, MBL was classified into the chronic lymphocytic leukemia (CLL) type (21.7%) and non-CLL-type (7.4%). Flow cytometry analysis and cell counts were used to determine the size of clonal population, and the data indicate that MBL and CLL are present in a continuous spectrum of clonal expansion. The findings may contribute to the current understanding of MBL and evaluation of incidental lymphocytosis. Further studies are required to evaluate clonal progression as a precursor stage of lymphoid malignancy.

To supplement and modify the diagnosis and clinical research of B-cell Chronic Lymphocytic Leukemia (B-CLL), a new method based on cell imaging and image processing was developed and applied to the B-CLL patient samples. The fluorophore-labelled leukemia cells were clearly visualized, reflecting the positive/negative expression of the corresponding surface markers and their distribution. Computer algorithms were devised and used to analyze a large number of images. The fluorescence intensity of the labelled antibodies on a given cell directly reflects the expression of the corresponding surface markers. The morphology and size of leukemia cells were not identical even in the same patient’s sample and the size variation does not correlate with the number of surface markers. The amount of each surface marker was approximately fixed for each patient, but there were some relationships, for instance, the number of CD19 and CD38 markers were correlated to each other. The heterogeneous expression of surface markers confirmed an assumption that surface markers have their preferred membrane positions. One of the most important results is that the cell imaging and our image processing method has provided an alternative and reliable way to diagnose B-CLL and new insights in the prognosis of subtype of B-CLL.

Monoclonal B lymphocytosis (MBL) has been recognized as a novel diagnostic condition. This study aims at the identification of clonal lymphocytosis in the patients with asymptomatic lymphocytosis. A total of 203 patients were evaluated for clonal B and T lymphocytosis by using flow cytometry and multiplex-PCR. Among them clonal B- or T-cells were detected in 54.2% of the cases, of which 38.4% were clonal B-cells and 15.8% were clonal T-cells cases. By immunophenotype, MBL was classified into the chronic lymphocytic leukemia (CLL) type (21.7%) and non-CLL-type (7.4%). Flow cytometry analysis and cell counts were used to determine the size of clonal population, and the data indicate that MBL and CLL are present in a continuous spectrum of clonal expansion. The findings may contribute to the current understanding of MBL and evaluation of incidental lymphocytosis. Further studies are required to evaluate clonal progression as a precursor stage of lymphoid malignancy.

Lymphocytes from normal subjects and patients with B-chronic lymphocytic leukemia (B-CLL) show functional responses to extracellular ATP characteristic of the P2X7 receptor. These responses include opening of a cation selective channel/pore which allows entry of the fluorescent dye, ethidium+ and activation of a membrane metalloprotease which sheds the adhesion molecule L-selectin. In this thesis, the surface expression of P2X7 receptors was measured in normal leucocytes, platelets and B-CLL lymphocytes and compared with their functional responses. Monocytes showed 4-5 fold greater expression of P2X7 than B-, T- and NK- lymphocytes, while P2X7 expression on neutrophils and platelets was weak. All cell types demonstrated abundant intracellular expression of this receptor. All 12 subjects with B-CLL expressed surface P2X7 at about the same level as for B-lymphocytes from normal subjects. P2X7 function, measured by ATP-induced uptake of ethidium, correlated closely with surface expression of this receptor in normal and B-CLL lymphocytes and monocytes. However, the ATP-induced uptake of ethidium into the malignant B-lymphocytes in 3 patients was low or absent. The lack of P2X7 function in these B-lymphocytes was confirmed by the failure of ATP to induce Ba2+ uptake into their lymphocytes. This lack of function of the P2X7 receptor resulted in a failure of ATP-induced shedding of L-selectin, an adhesion molecule which directs the recirculation of lymphocytes from blood into the lymph node. To study a possible genetic basis of non-functional P2X7 receptor, we sequenced DNA coding for the carboxyl terminal tail of P2X7. In 33 of 130 normal subjects a heterozygous nucleotide substitution (1513A--C) was found while 3 subject carried the homozygous substitution which codes for glutamic acid to alanine at amino acid position 496. Surface expression of P2X7 on lymphocytes was not affected by this 496Glu--Ala polymorphism demonstrated both by confocal microscopy and immunofluorescent staining. Monocytes and lymphocytes from the 496Glu--Ala homozygote subject expressed non-functional receptor while heterozygotes showed P2X7 function which was half that of wild type P2X7. Results of transfection experiments showed the mutant P2X7 receptor was non-functional when expressed at low receptor density but regained function at a high receptor density. This density-dependence of mutant P2X7 function was also seen on differentiation of fresh monocytes to macrophages with interferon-gamma which upregulated mutant P2X7 and partially restored its function. P2X7-mediated apoptosis of lymphocytes was impaired in homozygous mutant P2X7 compared with wild type. The data suggest that the glutamic acid at position 496 is required for optimal assembly of the P2X7 receptor. Apart from the 496Glu--Ala polymorphism, three other single nucleotide polymorphisms, 155His--Tyr, 348Ala--Thr and 568Ile--Asn were also found in the P2X7 receptor. The site directed mutant cDNA were generated for all 3 polymorphisms and transfected into HEK293 cells to study the impact of these polymorphisms on P2X7 function. Results suggested that Ile568 is important for P2X7 protein trafficking to cell surface. Further study of these two loss-of-function polymorphisms (496Glu--Ala and 568Ile--Asn) may help better understanding of the functional domains in the P2X7 receptor and its role in CLL, lymphoma and infectious diseases. Conclusions: 1.P2X7 receptor is expressed in human leukocytes, including lymphocytes, natural killer cells as well as monocytes, on both surface and intracellular locations. 2.Both the expression and function of P2X7 are highly variable between in human individuals. Non-functional P2X7 receptors are found in some subjects, including both normal subjects and CLL patients, and are often associated with defects in ATP-induced cytotoxicity and L-selectin shedding. 3.Two single nucleotide polymorphisms (SNPs), 496Glu--Ala and 568Ile--Asn, are found at low frequency in the human population and lead to the loss-of-function of P2X7. Both permeabllity function and the downstream effects mediated by P2X7 are affected by these two SNPs. The mechanisms for the loss-of-function differs between the two polymorphisms.

Resumo: Leucemia Linfóide Crônica (LLC) é uma neoplasia maligna de linfócitos B maduros com aspecto monomórfico, que se acumulam em tecidos linfóides secundários, medula óssea e sangue periférico. O diagnóstico é realizado com base em achados clínico-morfológicos e imunofenotípicos. Os pacientes com LLC são estratificados conforme o estadiamento clínico para definição de terapêutica; atualmente a descoberta de marcadores prognósticos, como ZAP-70, trouxe novas perspectivas ao tratamento, principalmente no estádio precoce. Os objetivos deste estudo foram: avaliar a eficácia da pesquisa da ZAP-70 em biópsia de medula óssea (BMO) e em inclusão do coágulo, utilizando-se a técnica de imuno-histoquímica; avaliar possíveis correlações deste marcador na BMO com evolução clínica e risco de progressão; correlacionar a imuno-expressão da ZAP- 70 com o arcabouço reticulinico das BMO; avaliar se este arcabouço possui correlação com o prognostico da doença e, por fim, traçar um perfil epidemiológico dos pacientes com LLC atendidos no Hospital das Clinicas da Faculdade de Medicina de Botucatu (FMB). Foram selecionados 153 pacientes com LLC, atendidos no ambulatório do Serviço de Hematologia da FMB-UNESP, de 1980 a 2008, e 9 pacientes assistidos no Serviço de Onco-Hematologia do Hospital Amaral Carvalho-Jaú, no período de 2000 a 2008, com o mesmo diagnóstico, perfazendo 162 casos. Destes, 79 possuíam prontuários que foram revisados, bem como dados morfológicos da BMO e AMO. Foi realizada a pesquisa da ZAP- 70 pela técnica de imuno-histoquímica nas amostras parafinadas de BMO e inclusão do coágulo. Observou-se que 55,7% dos pacientes eram do sexo masculino; 86,1% de etnia branca; a mediana de idade foi 65 anos; relação homem:mulher de 1,2:1. 40,5% dos pacientes tiveram diagnóstico por achado incidental e 73,4%, já com adenopatia secundária à admissão. 17,2% ...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Chronic lymphocytic leukemia (CLL) is a malignant neoplasm consisting of mature monomorphic B lymphocytes that accumulate in secondary lymphoid tissues, bone marrow and peripheral blood. Diagnosis is based on clinical, morphological and immunophenotypic findings. CLL patients are categorized according to clinical aspects for treatment management and currently, certain tools, such as prognostic marker ZAP- 70, have determined new treatment perspectives, especially for patients in early stages. The study aimed to evaluate the efficiency of ZAP-70 investigation in bone marrow biopsies (BMBs) and inclusion coagulate using immunohistochemistry; identify possible correlations of this marker in clinical evolution and risk of progression; correlate the immunoexpression of ZAP-70 with BMB reticulin network; evaluate whether this network correlates with disease prognosis and; outline an epidemiological profile of patients with CLL attended in the Clinics Hospital of Botucatu Faculty of Medicine (FMB). The sample consisted of 153 CLL patients attended at hematology outpatient clinic of the FMB-UNESP, from 1980 to 2008, and 9 CLL patients attended at the oncohematology service of the Amaral Carvalho Hospital, Jaú, from 2000 to 2008, totaling 162 cases. In 79 cases, medical records including BMB and bone marrow aspirate (BMA) morphological data were reviewed. Immunohistochemistry for ZAP-70 was conducted on paraffinated samples of BMB and inclusion coagulate. Observation revealed that: 55.7% were male, 86.1% white, with a median age of 65 years-old; 40.5% of cases were diagnosed by incidental findings in routine blood smears; 73.4% were diagnosed with peripheral adenomegaly on admission. 17.2% were diagnosed in the early stages and 15% of cases showed prolonged remission. BMB and BMA morphology revealed that 94.8% of patients presented increased cellularity... (Complete abstract click electronic access below) / Orientador: Lígia Niéro-Melo / Coorientador: Maria Aparecida Custódio Domingues / Banca: Lucilene Silva Ruiz e Resende / Banca: Lisandro Ferreria Lopes / Mestre

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / A Leucemia LinfocÃtica Aguda (LLA) e o Linfoma nÃo-Hodgkin (LNH) estÃo entre os mais frequentes tipos de neoplasias em crianÃas. A prevenÃÃo e controle do cÃncer devem ser priorizados, tendo em vista sua alta prevalÃncia e crescente relevÃncia como causa de morte em muitos paÃses, alÃm do grande volume de recursos financeiros consumidos. O nordeste brasileiro à pobre em estudos epidemiolÃgicos sobre o cÃncer infantil. O objetivo deste estudo foi traÃar o perfil dos pacientes portadores de LLA e LNH admitidos no perÃodo de 2001 a 2005 no Hospital Infantil Albert Sabin. Estudo observacional, descritivo e retrospectivo. 325 prontuÃrios (254 casos de LLA e 71 de LNH) de pacientes entre 0 e 18 anos e 11 meses foram revisados. Os dados foram inseridos em bancos de dados dos programas SPSS 14.0, Epi Info 3.3.2 e Microsoft Excel 2007. O ponto de corte para desnutriÃÃo foi o escore Z igual a -2 desvios-padrÃo. Na anÃlise estatÃstica foram utilizados o teste exato de Fisher, Qui-quadrado, Student (t), Mann-Whitney, Shapiro-Wilk, Levene, Log-rank, modelo de regressÃo de Cox e mÃtodo de Kaplan Meier para anÃlise de sobrevida. O nÃvel de significÃncia foi p<0,05. A populaÃÃo deste estudo constituiu-se predominantemente por pacientes do sexo masculino (63,4%), faixa etÃria de 02 a 06 anos (49,8%), cor nÃo-branca (62,5%); provenientes da capital e regiÃo metropolitana (56,9%) e com prognÃstico de alto risco (59,1%). 38,3% evoluÃram a Ãbito. As principais manifestaÃÃes clÃnicas iniciais foram febre, anemia, emagrecimento e cansaÃo nos casos de LLA; e febre, massa tumoral palpÃvel, anemia e dor abdominal nos casos de LNH. O tempo mÃdio de duraÃÃo das queixas foi de 3,9 meses. CrianÃas de 0-1 e de 13-18 anos apresentaram pior prognÃstico. A cor da pele, o prognÃstico, o protocolo de tratamento e os sintomas/sinais iniciais febre, cansaÃo e vÃmito mostraram associaÃÃo significativa em relaÃÃo aos Ãbitos. Os protocolos terapÃuticos mais utilizados foram adaptados do LLA 93 e LNH 95. 31% dos pacientes em uso do LLA 93 e 49,2% em uso do LNH 95 evoluÃram a Ãbito. Maior percentual de Ãbito ocorreu no grupo de alto risco (56,94%) e durante a fase de induÃÃo (36,11%) do protocolo LLA 93. Entre os pacientes de baixo risco, 39,28% faleceram durante a fase de manutenÃÃo e 17,85% apÃs o fim do protocolo. 48,78% dos pacientes de alto risco faleceram durante a fase de induÃÃo. Entre os 25 casos que utilizavam o protocolo LNH 95 e faleceram, 4% correspondiam a Linfoma de alto risco oriundos de cÃlulas T e 96% de cÃlulas B. 53,31% (n=15) dos pacientes com Linfoma de cÃlulas B e risco intermediÃrio para recaÃda faleceram durante o Ciclo A do tratamento. Nove pacientes apresentavam alto risco para recaÃda e 33,34% faleceram na fase de CitorreduÃÃo. O Ãndice de desnutriÃÃo para os pacientes com LLA foi de 8,3%, 6,0% e 5,6% e para LNH foi 12,3%, 14,1% e 15,9% em relaÃÃo a peso/estatura, peso/idade e estatura/idade, respectivamente. Pacientes com LLA apresentaram dÃficit maior no Ãndice peso/estatura, indicativo de um processo de desnutriÃÃo aguda. Maior dÃficit no Ãndice estatura/idade entre os pacientes com LNH indica um processo de desnutriÃÃo crÃnica. Os resultados acerca da frequÃncia dessas patologias, faixa etÃria e sexo foram equivalentes aos encontrados na maioria dos estudos. O prognÃstico inicial e o protocolo terapÃutico indicam uma possÃvel influÃncia sobre o desfecho do tratamento. Estudos adicionais sÃo necessÃrios para avaliar a influÃncia da quimioterapia, cor da pele, estado nutricional e outros fatores sobre o tempo de sobrevida do paciente com cÃncer. Os profissionais de saÃde e a populaÃÃo leiga precisam conhecer melhor e estar atentos Ãs manifestaÃÃes clÃnicas iniciais das neoplasias a fim de facilitar o diagnÃstico precoce. / The Acute Lymphocytic Leukemia (ALL) and the Non-Hodgkin Lymphoma (NHL) are among the most frequent types of cancer in children. The prevention and control of the cancer must be prioritized, in view of its high prevalence and increasing relevance as cause of death in many countries, beyond the great sum of consumed financial resources. The Brazilian northeast is poor in epidemiological studies about cancer in the children. The objective of this paper was to set the profile of the patients with LLA and LNH admitted in the Hospital Infantil Albert Sabin between 2001 and 2005. Observational descriptive and retrospective study. 325 medical registers (254 cases of LLA and 71 of LNH) of patients among 0 and 18 years and 11 months had been revised. The data had been inserted in data bases of the programs SPSS 14.0, Epi 3.3.2 Info and Microsoft Excel 2007. A Z-score cut-off point of <-2 SD was used to classify the malnutrition. The Fisherâs Exact Test, Qui-square, Student (t), Mann-Whitney, Shapiro-Wilk, Levene, Log-rank, Cox regression and Kaplan Meier Survival Probability Estimates were used in the statistical analyses. The level of significance was p<0,05. The population of this study was predominantly male (63.4%), 02 to 06 years age-group (49.8%), non-white (62.5%), from the capital and metropolitan region (56.9%) and with prognostic of high risk (59.1%). 38.3% died. The main clinical manifestations had been fever, anaemia, loss of weight and fatigue in the LLA cases; and fever, anaemia, palpable tumor mass and abdominal pain in the LNH cases. The mean duration time of the complaints was 3.9 months. Children with 0 to 1 and 13 to 18 years had presented worse prognosis. The color of the skin, the prognosis, the treatment protocol and the initial clinical manifestations (fever, fatigue and vomit) had shown significant association in relation to the deaths. The therapeutical protocols more used were adapted of the LLA 93 and LNH 95. 31% of the patients in use of LLA 93 and 49.2% in use of LNH 95 died. The largest percent of deaths were in the group of high risk (56.94%) and in the induction phase (36.11%) of protocol LLA 93. Between the patients of low risk, 39.28% died during the maintenance phase and 17.85% after the end of the protocol. 48.78% of the patients of high risk died during the induction phase. Among the 25 cases that used protocol LNH 95 and died, 4% corresponded to lymphoma of high risk deriving of the cells T and 96% of the cells B. 53.31% (n=15) of the patients with lymphoma of the cells B and intermediate risk to fallen died during the Cycle A of the treatment. Nine patients presented high risk for fallen and 33.34% died in the cytoreduction phase. The malnutrition indices to the LLA patients were of 8.3%, 6.0% and 5.6% and to LNH were 12.3%, 14.1% and 15.9% in relation the weight/height, weight/age and height/age, respectively. Patients with LLA had presented larger deficit in the index weight/height, indicative of a process of acute malnutrition. Larger deficit in the height/age index between the patients with LNH indicates a process of chronic malnutrition. The results about the frequency of these disease, age-group and gender were equivalents to those encountered in the majority of studies. The initial prognosis and the therapeutical protocol indicate an influence on the outcome of the treatment. Other studies are necessary to evaluate the influence of the chemotherapy, color of the skin, nutritional status and other factors on the survival time of the patient with cancer. The professionals of health and the laypeople need to know better and to be intent to the initial clinical manifestations of the neoplasm disease in order to facilitate the precocious diagnosis.

Magister Scientiae (Nutrition Management) - MSc(NM) / Acute lymphocytic leukemia (ALL) is the most common type of childhood cancer accounting for approximately 25% of cancers diagnosed in children less than 20 years of age. It originates in the bone marrow and prevents the normal manufacture of red blood cells, white blood cells and platelets. A poor nutritional status is frequently observed in children with ALL at the time of diagnosis and during treatment which may result in protein energy malnutrition if nutrition intervention is delayed. This retrospective study aims to assess the nutritional status of children newly diagnosed with Acute Lymphoblastic Leukemia (ALL) using 20 case studies between 1 January 2013 and 31 December 2014 from Sheikh Khalifa Medical City (Abu Dhabi, UAE), in order to develop an appropriate nutritional support protocol for pediatric ALL patients treated at this institution. Study Design: A retrospective descriptive case study design was used. The study population consisted of 20 electronic medical records of patients aged between 1-14 years who were newly diagnosed with Acute Lymphoblastic Leukemia (ALL) and admitted to Sheikh Khalifa Medical City for treatment during the period 1 January 2012 and 31 Dec 2014. Data Collection: Identification of suitable participants began through a review of each potential study participant`s electronic medical record. Data was collected and recorded on a data collection form (Appendix III) from the electronic medical record for each suitable participant for the following at admission and during the full duration of all phases of cancer treatment namely induction, consolidation, interim maintenance, delayed intensification and maintenance. The data collected comprised of the following: age, gender, date of diagnosis, symptoms on diagnosis, the cancer diagnosis (type and subtype), anthropometric measurements (weight, length/ height, head circumference), biochemical values (visceral proteins, blood glucose levels, hemoglobin, hematocrit, lymphocyte count), clinical assessment (stomatitis, anemia, mucositis), diet history (home feeding regimes; consumption of daily requirements; food preferences – types, textures; food allergies, food intolerances; food aversions; use of oral nutritional supplements; treatment-related side-effects; systemic related side-effects (nausea; vomiting; diarrhea; anorexia; appetite changes; taste changes; physical activity level; depression), dietary requirements (age and gender related nutritional requirements for energy, protein, fat and fluids) and indications for nutritional support (oral feeding; enteral feeding; parenteral feeding). Analysis of Results: The weights and length/ heights of participants recorded in the electronic medical records were converted to z-scores on the World Health Organization growth charts. The diet prescription of nutritional intervention was interpreted in comparison to the biochemical indices, anthropometric status and dietary intake of each participant. All the data involving changes in anthropometrics, biochemistry, diet history and nutritional interventions from each case study (from diagnosis and through all stages of treatment) was screened and compared with reference values in the context of the age and sex of the child. Evidence based nutritional guidelines were used to document the outcomes of the medical nutrition treatment provided in order to develop a nutrition support protocol for children with Acute Lymphoblastic Leukemia at Sheikh Khalifa Medical City. Results: The results showed that weight loss expressed as a percentage of body weight provided a more accurate estimate of the true significance of weight loss in subjects undergoing cancer treatment (chemotherapy) for ALL. A weight loss of greater than 5% of body weight over a period of one month is considered a sign of nutritional deprivation even if the subject is not classified as undernourished by anthropometric parameters. Subjects experienced the highest weight loss during the consolidation phase and interim maintenance phases of treatment. Conclusion: It can therefore be concluded that pediatric subjects on cancer treatment for ALL at SKMC and receiving nutritional support underwent changes in nutritional status as manifest by a reduction in more than 5% of their body weight during three phases of treatment namely induction, consolidation and interim maintenance. An appropriate nutrition support protocol was developed based on the results and experience obtained from this study for pediatric ALL patients treated at SKMC.

The estrogen receptor alpha (ERa) and estrogen receptor beta (ER~) have been demonstrated to be important for immune system regulation and studies have suggested an antiproliferative effect of ER~ in lymphoid malignancies. We have studied the expression of ERa and ER~ in peripheral blood mononuclear cells from patients with chronic lymphocytic leukemia (CLL). Expression of ERa was low, while ER~ was highly expressed in CLL cells. In order to investigate the possible inhibitory effect of ligand-activated ER~ , we treated CLL cells and Mecl cell lines with the selective ER~ agonist diarypropionitrile (DPN) in culture. Treating Mecl cell lines with DPN showed an antiproliferative effect of ER~ agonist by significantly inhibit the growth of Mec 1 cell lines. This suggests that ER~ agonist may be useful in the treatment of CLL.

Chronic lymphocytic leukemia (CLL) cells are considered to be derived from antigen-exposed B cells. To further explore the antigen-driven selection behind the leukemogenesis of CLL, we performed immunoglobulin (Ig) specificity screening of 7 CLL cell lines and 23 primary CLL clones from patient peripheral blood. We also included a recombinant monovalent monoclonal antibody (mAb) belonging to a subset of CLL cases with identical or semiidentical heavy chain complementarity determining region 3 (HCDR3) of the IGHV3-21 gene rearrangement. We found CLL mAb specificities against vimentin, filamin B, cofilin-1, proline-rich acidic protein 1, cardiolipin, oxidized low density lipoprotein and Streptococcus pneumoniae polysaccarides. These molecules are functionally associated with microbial infection and/or apoptotic cell removal. An antigen-driven selection would therefore imply that CLL B cell precursors are involved in the elimination and scavenging of pathogens and apoptotic cells, which could trigger the development of the disease. The limited in vitro survival of CLL cells makes Epstein-Barr virus (EBV) immortalization of CLL cells a useful experimental model for studies on antibody-specificity screening. Considering the intricate procedure of EBV transformation of CLL cells and the many false cell lines used worldwide, we also wanted to characterize and evaluate the authentic origin of several previously established CLL cell lines and their normal lymphoblastoid counterparts. Three of the CLL cell lines tested were truly authentic (I83-E95, CLL-HG3 and CII), two had features of a biclonal Ig expression (232B4 and WaC3CD5+), one was only tentatively verified (PGA-1), whereas one cell line could not be verified (EHEB) due to lack of original patient cells for comparison. Two of the presumed normal lymphoblastoid cell lines tested were shown to be a neoplastic CLL clone. This study emphasizes the importance of proper cell line authentication and we will continue to verify additional cell lines not yet proven authentic. In conclusion, we provide evidence for natural Ab production by CLL cells and suggest that these cells might be derived from B cell precursors involved in the innate immunity and, thus, providing a first-line-defence against pathogens and in elimination of apoptotic cells.