Day 1 :
Keynote Forum
C D Atreya
Center for Biologics Evaluation and Research, Food and Drug Administration, USA
Keynote: Small regulatory RNAs in blood component storage and hemophilia A disease manifestation
Time : 10:00-10:40
Biography:
Abstract:
Discovery of small noncoding RNAs (e.g. microRNAs) have revolutionized the field of cell biology with the seminal discovery that that these tiny molecules are the gene regulators impacting almost all aspects of the cellular processes in human health and disease. MiRNAs are small (17-22 nucleotides) single-stranded noncoding RNAs that bind to their specific target sites on mRNAs via Argonaute-2 (AGO-2) protein-associated miRNA-induced silencing complex (RISC). Such functional complexes have been identified in a variety of cells. Blood cells stored for transfusion undergo several physiological changes and several miRNAs were identified to be differentially expressed during platelet storage that either can serve as predictors of poor quality of the stored product or, regulators of cellular processes. Recently it was identified in stored platelets that miR-570 interacts with and down regulates the expression of mitochondrial ATPase subunit g (ATP5L) encoding mRNA and several miRNAs associated with AGO-2 in stored red blood cells (RBC) suggesting their role in RBC both in vivo and in storage. MicroRNAs also play a role in hereditary blood disorders such as hemophilia A (HA). It has been shown that miR-1246 expression is high in HA patients and was inversely correlated with factor VIII expression in cell culture. Subsequently several miRNAs have been identified in HA patients that have the potential to serve both as biomarkers and regulators of HA disease phenotype. Overall, the advances made in the small regulatory RNA field with regards to stored cellular blood components and hemophilia A clearly suggest that these RNAs are critical players that needs to be studied to develop strategies to enhance the quality of stored blood components and to improve therapies for HA patients.
Keynote Forum
Chava Kimchi-Sarfaty
Center for Biologics Evaluation and Research, Food and Drug Administration, USA
Keynote: New platform technologies for biotherapeutics and emerging rewards and risks
Time : 10:40-11:20
Biography:
Abstract:
Therapeutic proteins now represent a growing and critical component of drug development. Biotherapeutics such as antibodies, recombinant enzymes, gene and cell therapies continue to revolutionize the treatment of many diseases. The production and regulatory approval of these complex molecules are not without their own inherent challenges. A growing list of protein engineering strategies exists to improve the circulating half-life, bioavailability, function and stability of recombinant protein therapeutics. To aid in biomanufacturing, protein therapeutics are often strategically designed with synonymous and/or non-synonymous mutations to eliminate rate-limiting transcriptional/translational features in the native expression sequence. In a technique known as codon optimization, hundreds of synonymous mutations are strategically incorporated into the expression vectors of recombinant therapeutic proteins. We seek to understand how protein biogenesis can be modulated by genetic variants in the coding sequence using model recombinant proteins regulated by CBER. To this end, we have evaluated existing and new laboratory assays that are sensitive to transcriptional, translational and posttranslational processes. These emerging technologies for recombinant protein design are regularly evaluated at FDA’s CBER, which encompasses a holistic view of protein therapeutic development and monitors the process from the bench to the bedside to ensure effective development and product licensing. CBER’s goals include development and enforcement of laboratory standards by evaluating technologies and reagents as well as the assessment of pre-clinical models and clinical approaches. These goals are in line with CBER’s vision to improve the safety of biological products and ultimately advance public health.
- Hematology | Blood Disorders | Hematology and Oncology
Location: Johnson
Chair
C D Atreya
Center for Biologics Evaluation and Research, Food and Drug Administration, USA
Session Introduction
Chava Kimchi-Sarfaty
Center for Biologics Evaluation and Research, Food and Drug Administration, USA
Title: Assay-dependent results of ADAMTS13 activity in sickle cell disease
Biography:
Abstract:
Nahla Osman
Menoufia University, Egypt
Title: Higher mTOR mRNA is associated with poor outcome in AML patients
Biography:
Abstract:
Neetu Dahiya
Food and Drug Administration, USA
Title: Study of miRNAs and mRNAs in stored platelets identifies the potential functional pathways relevant to platelet storage lesions
Biography:
Neetu Dahiya has expertise in understanding cellular gene regulation by small noncoding RNAs such as microRNAs (miRNAs). Her recent work is focused on the changes in expression of both miRNAs and mRNAs in platelets during storage. Some of these studies already identified miRNA:mRNA interactions that demonstrate the role of miRNAs in platelet biology during ex vivo storage and the results and concepts have been published in peer-reviewed journals.
Abstract:
Shahtaj Khan
Hayatabad Medical Complex hospital, Pakistan
Title: Pattern of adulthood hematological malignancies in Khyber Pakhtunkhwa
Biography:
Shahtaj Khan is an Assistant Professor of Hematology and Head of the Department of Pathology at Hayatabad Medical Complex, Peshawar, Pakistan. She is also working as Consultant Hematologist at Rehman Medical Institute. Her research interests reflect in her wide range of publications in various national and international journals.
Abstract:
Objective: To evaluate the frequency of adulthood hematological malignancies in Khyber Pakhtunkhwa population.
Material & Method: This descriptive observational study was carried out at Diagnostic Laboratory Rehman Medical Institute (RMI), and Hayatabad Medical Complex, Peshawar Pakistan, from December 2014 to December 2017. A total of 571 adult patients suspected to have hematological malignancies were included in the study. All these patients were examined in clinics by different physicians and referred to pathology department for bone marrow aspiration and trephine biopsy. Two millilitre of peripheral blood was collected in EDTA vacutainer tube and complete blood count, retic count along with peripheral film examination was done. Bone marrow aspiration and trephine biopsy samples were taken from all the patients. Aspiration and trephine biopsy slides were examined and further immunohistochemistry and flow cytometry was done for complete diagnosis. All the data was recorded, analyzed and presented in tables.
Results: Out of the 571 cases referred for bone marrow examination. 259 adult patients were diagnosed with different types of hematological malignancies. Out of 259, 186 (71.8%) were males and 73 (28.2%) were females. The age range of studied population was from 18 to 84 years with average age of 46.21 years. Out of them, 96 (37.1%) were diagnosed with myeloid hematological malignancies and 163 (62.9%) were diagnosed with lymphoid hematological malignancies. Acute myeloid leukemia (22.3%), acute lymphoblastic leukemia (21.6%) and chronic lymphocytic leukemia (18.9%) were more prevalent hematological malignancies in this region while plasma cell leukemia, polycythemia rubra vera and hairy cell leukemia were least common hematological malignancies. The frequencies of other hematological malignancies were lymphoma (10.4%), multiple myeloma (9.7%), chronic myeloid leukemia (7.3%), primary myelofibrosis (2.7%), myelodysplastic syndrome (2.7%) and essential thrombocythemia (1.1%) in total hematological malignancies.
B Yusuf Jamoh
Ahmadu Bello University Teaching Hospital, Nigeria
Title: Soluble CD36 as a determinant of disease severity among patients with sickle cell anaemia in Nigeria
Biography:
B Yusuf Jamoh has completed his MBBS programme from Bayero University, Kano, Nigeria and had MSc Cancer Biology, with commendation, from Kingston University, London. He is a Fellow of National Postgraduate Medical College of Nigeria and was appointed as Honorary Consultant Physician, Ahmadu Bello University Teaching Hospital (ABUTH), Zaria, Nigeria. He is the Head of Clinical Haematology Unit, ABUTH. He has published 12 papers in reputed journals and he is currently acting Postgraduate Coordinator, Department of Medicine, Ahmadu Bello University, Zaria, Nigeria.
Abstract:
Background: This descriptive, cross-sectional study was aimed at determining the relationship between soluble CD36 (sCD36; a cell adhesion marker), levels of % Hb F, haematological parameters and disease severity in adults with SCA in Kano, Nigeria.
Methods: One hundred and forty subjects with SCA in steady state were purposively selected and compared with 70 apparently healthy controls. Ten milliliters of venous blood was obtained to determine CBC (using auto haematology analyzer), % Hb F (estimated by modified Betke’s method) and sCD36, using Human Soluble CD36 Elisa Kit (ADIPO Bioscience Inc., USA). Severity was assessed by El-Hazmi’s scoring system. Student t-test and Pearson’s correlation were used as statistical test and P-value ≤ 0.05 was used to define statistical significance.
Results: The median sCD36 was significantly higher (P<0.01) in subjects with SCA (22.3 ng/ml) than in the controls (14.8 ng/ml). A direct correlation was observed between sCD36 and WBC count (ρ=0.7410; P<0.001), an inverse correlation was observed between sCD36 and % Hb F (ρ=-0.5406; P<0.001) and a direct correlation was observed between sCD36 and severity score (ρ=0.5808; P<0.001) in the subjects. No such relationship was observed among the parameters in the control group. Complications like ACS, stroke, retinopathy and AVN of the femoral head were observed to be associated with high sCD36 levels. A multiple logistic regression modeling revealed that WBC count predicted the most significant odds (OR=3.87; P<0.001) for sCD36 positivity.
Conclusion: The level of sCD36 is a marker of disease severity and may predict the occurrence of vascular-related complications of adults with SCA; and WBC alone may be used as a surrogate marker of sCD36 level.
- Leukaemia | Hematology Oncology
Location: Johnson
Chair
Iman Atteya eltounsi
Menoufia University, Egypt
Session Introduction
Haytham Dahlawi
Taif University, Saudi Arabia
Title: Impact of anthocyanin chemical structure found in pomegranate juice on leukaemia treatment
Biography:
Haytham Dahlawi has completed his PhD from Sheffield Hallam University. He is the Vice-Dean and Head of Exam Committee of College of Applied Medical Sciences, Taif University. He has published more than eight papers. He is interested in polyphenols and their potential effect on treatment of blood.
Abstract:
Anthocyanins are an abundant group of flavonoids present in pomegranate juice. Anthocyanidins lack the sugar component of the parent anthocyanin. Anthocyanins are the mono and diglycosylated forms of anthocyanidins with substitutions at the 3 and 5 positions. The six most abundant anthocyanins in pomegranate juice are cyanidin-3-O-glucoside, cyanidin-3,5- di-O-glucoside, delphinidin-3-O-glucoside, delphinidin-3,5-di-O-glucoside, pelargonidin-3-O-glucoside and pelargonidin- 3,5-di-O-glucoside. Anthocyanins and anthocyanidins have demonstrated cancer chemopreventive properties in breast, skin, lung and gastrointestinal carcinogenesis. In this study we investigated the effect of anthocyanin chemical structure found in pomegranate juice on leukaemia cells. The anti-cancerous effect of eight different anthocyanins was investigated on four leukaemia cell lines (CCRF-CEM, MOLT-3, HL-60 and THP-1). Cells were treated with 0 μM to 100 μM anthocyanins for 24 hours. Cell proliferation was assessed using CellTiter-Glo® luminescent cell viability assay. The pro-apoptotic actions of anthocyanins were assessed by two assays: Annexin V/Propidium iodide staining and staining for caspase3-activity using flow cytometry. Delphinidin was found to have the greatest inhibitory effect on cell proliferation which was found to be significantly greater than that shown by cyanidin and pelargonidin. Delphinidin also significantly induced apoptosis in all four cell lines. Cyanidin induced apoptosis only in CCRF-CEM and pelargonidin failed to induce apoptosis in any cell lines. Anthocyanins containing sugar molecules showed decreased toxicity which correlated with the size of sugar molecule. These results provide evidence that anthocyanins show anti-cancer effects which are dependent on chemical structure and association with sugar molecules.
Mario da Silva Garrote Filho
Federal University of Uberlândia, Brazil
Title: Influence of resistance training on erythrocyte stability and hematologic, and biochemical variables in breast cancer survivors
Biography:
Mario da Silva Garrote Filho completed his PhD from Federal University of Uberlândia, and is currently developing Postdoctoral studies at the same university. He has published more than 15 articles in renowned journals. He also develops interactive animations in Animate CC and has great knowledge in Statistics, with special abilities in the use of SPSS and Excel.
Abstract:
The objective of this study was to investigate the effects of 12 weeks of resistance training (RT) on hematologic, biochemical and erythrocyte membrane stability variables in a population (n=14) of breast cancer survivors (BCS). Blood collections and laboratory tests were performed before and after the training period. The RT program contributed to the promotion of significant declines in triglycerides and total and LDL cholesterol, and a significant elevation in HDL-cholesterol. There were also significant declines in erythrocytes count, although values have still remained within the reference range of these variables. The observed decline in the RBC count was associated with increasing levels of HDL C and decreasing levels of LDL-C, with no association with changes in the erythrocyte stability variables. This is immensely relevant and should mean that the reduction of lipidemia should not be seen as an isolated goal, outside the hematologic context. The results also shows the importance of monitoring any type of treatment of BCS, even physical exercise, with periodic hematologic and biochemical evaluations.
Viola Maria Popov
Colentina Clinical Hospital, Romania
Title: The analyses of influence of platelet membrane fluidity in platelet receptors expression in chronic myeloproliferative neoplasms patients
Biography:
Viola Maria Popov has completed his PhD from Carol Davila University Bucharest. He is the project Manager of one research project - the role of microparticles in thrombotic complication in chronic myeloproliferative neoplasms patients. He has published more than 10 papers in journals A and B+
Abstract:
Background: Patients with chronic myeloproliferative neoplasms (MPNs) had qualitative and quantitative modifications of platelet membrane receptors that are involved in alteration of platelet function. These modifications of platelet functions determined thrombotic complications. The aim of our study was to determine if changes of platelet membrane fluidity could be correlated with alterations in expression of platelet receptors.
Materials & Methods: This retrospective study included 60 patients with MPNs as well as 10 patients with thrombosis not associated with MPNs. The group of patients with myeloproliferative neoplasms included 12 patients with chronic myeloid leukemia and 38 patients with MPNs Ph negative (essential thrombocythemia, polycythemia vera and idiopathic myelofibrosis). Depending on thrombosis presence in medical history of patients enrolled, we divided the MPNs patients into two groups: patients without thrombosis (50 patients) and patients with thrombosis in their medical history (stroke, myocardial infarction and splanchnic thrombosis). The determination of platelet membrane fluidity was performed by fluorescence anisotropy measurements using as marker 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene p-toluenesulfonate (TMA DPH). Fluorescence anisotropy (FA) was analysed depending the expression of platelet membrane receptors measured by flow cytometry analyses. We examined the flow cytometry markers of platelet adhesion (CD42a, CD42b), aggregation (CD41, CD61) and CD36.
Results & Discussion: The expression of CD41 receptor was lower in MPNs group (min 28.58, max 100.75) compared with controls group (min 43.58, max 89.46), p=0.02. We did not obtain statistical difference between expressions of CD61, CD42a, CD42b and CD 36. The fluorescence anisotropy of platelet membrane in MPNs group is higher than control group, p=0.05 The CD36 expression had a positive correlation with value of fluorescence anisotropy in MPNs patients group. (rho=0.75-95%; CI 0.308-0.925; p=0.005). We did not obtain any correlations between the rest of platelet receptors and FA modification.
Conclusions: MPNs patients have a lower membrane fluidity corresponding to higher FA. The expression of CD36 is higher in MPNs patients and was direct correlated with FA modifications. Platelet receptor CD36 recognizes oxidized lipids in oxidized low-density lipoprotein (oxLDL) particles, a process that determines the thrombosis process. The direct role of CD36 in thrombosis process has to be determined.
Haytham Dahlawi
Taif University, Saudi Arabia
Title: Prevalence of hemoglobinopathies in Taif city, Saudi Arabia
Biography:
Haytham Dahlawi has completed his PhD from Sheffield Hallam University. He is the Vice-Dean and Head of Exam Committee of College of Applied Medical Sciences, Taif University. He has published more than eight papers. He is interested in polyphenols and their potential effect on treatment of blood.
Abstract:
Hemoglobin variants can be either hemoglobinopathies which are responsible for diseases or non-pathological variants which couldn’t make any detectable disorder. Carriers with structural variant haemoglobin have 30 to 50% of the variant haemoglobin in their red blood cells. The most common variant haemoglobin is hemoglobin S, which accounts for 40% of carriers and responsible for more than 80% of disorders related to hemoglobinopathies. According to the WHO there are at least 948,000 new carrier couples, and over 1.7 million pregnancies to carrier couples every year. Thus, it is very important to provide a systematic carrier screening program specially among at-high risk couples. This might help to prevent or/and reduce the incidence of blood disorders that related to variant haemoglobin. In this study, a total of 9008 blood samples were studied among Saudi male and female, who attending to the centre of premarital screening at Taif city. Of these, 118 cases were displayed abnormal hemoglobin fractions on HPLC. The result of this study showed that Hb S heterozygous was presented as the major abnormality with 58.5% followed by beta thalassemia minor with 21%. Clear understanding the genetics and the prevalence of these diseases will provide opportunities for prevention or and reduce the incidence. Thus, this study suggests that in addition to the huge efforts already accomplished by the Saudi Ministry of Health to prevent at-risk marriages, the early diagnosis for these disorders might be offered for young adults as they can discuss the issue in the early stage of the marriage proposal.
Erhabor Osaro
Usmanu Danfodiyo University, Nigeria
Title: Some haematological parameters and ascorbate deficiency among children of African descent with protein energy malnutrition in Sokoto, Nigeria
Biography:
Erhabor Osaro is a Professor of Haematology, Transfusion Medicine and Laboratory Total Quality Management. He received his PhD in Immuno-Haematology from the Rivers State University of Science and Technology in Port Harcourt Nigeria. He is also an Alumni of University of Greenwich in the United Kingdom and Francis Tuttle College of Technology in Oklahoma, USA. Currently, he is working as Professor in Usmanu Danfodiyo University, Sokoto, Nigeria. His experience spans both Africa and Europe. He is on the Science Council of the United Kingdom register as a Chartered Scientist. He is Fellow and an Examiner (Registration Portfolio Verifier) for the Institute of Biomedical Science of London. He is the author of 4 scientific books and 6 chapters of scientific books.
Abstract:
Protein energy malnutrition is the most widespread nutritional deficiency disorder of mankind and continues to be a major public health burden in developing countries. The aim of this case-control study was to determine the changes in some haematological parameters and ascorbic acid levels among children of African descent with PEM in Sokoto, North Western Nigeria. The study included a total of 90 children (47 subjects with protein energy malnutrition and 43 apparently healthy controls) aged 6 months - 5 years, admitted to the Pediatric Units of Usmanu Danfodiyo University Teaching Hospital and Specialist Hospital, Sokoto. Some haematological parameters (packed cell volume, total white blood cell count and platelet count) were analyzed using the auto-hematology analyzer (Genesis, HA6000). Ascorbic acid levels were assayed by a standard chemical method. Nutritional status was determined using the Wellcome Trust Classification. Data were analyzed using SPSS 22.0 statistical package. A p-value ≤ 0.05 was considered significant in all statistical comparisons. The result indicated that subjects with protein energy malnutrition had a lower mean packed cell volume (25.50±0.996%) compared to controls (32.73±1.004%) (p=0.0001). The mean total white cell count was significantly higher among subjects with protein energy malnutrition (12.16±0.72x109/l) compared to controls (7.59±0.49x109/l) (p=0.0001). There were no statistically significant differences in the mean value of platelet counts of subjects (260.40±21.71x109/l) and controls (237.61±15.13x109/l) (p=0.400). The mean value of ascorbic acid was significantly lower among subjects (0.82±0.04 mg/dl) compared to controls (1.06±0.02 mg/dl) (p=0.0001). Children with kwashiorkor had higher value of packed cell volume compared to those with marasmickwashiorkor (p=0.0001). Children with marasmic-kwashiorkor had a higher total white cell count when compared with other types of protein energy malnutrition (p=0.0001). Underweight subjects had lower ascorbic acid levels when compared with other types of protein energy malnutrition (p=0.0001). Platelet count showed no significant difference within the various types of protein energy malnutrition (p=0.331). This study has shown that children with protein energy malnutrition have lower packed cell volume and ascorbic acid levels compared to controls. The total white cell count was higher among children with protein energy malnutrition compared to controls. Protein energy malnutrition was more prevalent among children from low socioeconomic class whose mothers have no formal education. Marasmus was the most common type of protein energy malnutrition. Children with kwashiorkor have a higher packed cell volume compared to other types of protein energy malnutrition. Total white blood cell count of children with marasmic-kwashiorkor was significantly higher compared with other types. Immune boosters (vitamins and other micronutrient) should be provided for school children particularly children with protein energy malnutrition. There is need for infant feeding practice to be strengthened by promoting exclusive breast feeding. There is need for increased enrollment of women in schools, enlightenment on nutritional education and empowerment so as to improve their socioeconomic status.