Leukemic stem cells shall be searched in the bone marrow before "tyrosine kinase inhibitor-discontinuation" in chronic myeloid leukemia.

BACKGROUND: Leukemic stem cells (LSCs) of chronic myeloid leukemia (CML), persisting in the bone marrow (BM) niche, could be responsible for the relapses within the patients of whom the treatment-free remission (TFR) had been attempted. We assessed the presence of the CML LSCs in the peripheral blood (PB) and concurrently in the BM in the patients with chronic-phase CML (CP CML). PATIENTS AND METHODS: Thirty-eight patients with CP CML were included into the study. CD45+ /CD34+ /CD38- cells with positive CD26 expression were considered as CML LSCs (CD26+ LSC) by using multiparameter flow cytometry (FCM). RESULTS: Mean BCR-ABL, PB LSC, and BM LSC were 58.528 IS (37.405-83.414 IS), 237.5 LSC/µL (16-737.5 LSC/µL), and 805 LSC/106  WBCs (134.6-2470 LSC/106  WBCs), respectively, in newly diagnosed CML patients. In the patients with BCR-ABL positive hematopoiesis, mean BCR-ABL, PB LSCs, and BM LSCs were 30.09 IS (0.024-147.690 IS), 13.5 LSC/µL (0-248.7 LSC/µL) and 143.5 LSC/106  WBCs (9-455.2 LSC/106  WBCs), respectively. No CML LSCs were detected in PB of patients who achieved deep molecular response (DMR). BM LSCs of the patients who were in DMR were 281.1 LSC/106  WBCs (3.1-613.7 LSC/106  WBCs). The amount of PB LSCs was highest in patients with newly diagnosed CML (P < .001). CONCLUSION: LSCs persisted in the BM of the patients with DMR, whereas there was no LSCs in the peripheral blood. The investigation of the CML LSCs in bone marrow before deciding TKI discontinuation could be justified to achieve and maintain stable TFR.

Protein Kinases in Hematological Disorders.

Cell signaling is an important part of the complex system of molecular communication that governs basic cellular activities and coordinates cell cycle machinery. Pathological alterations in the cellular information processing may be responsible for the diseases such as cancer. Numerous diseases may be treated effectively via the pharmacological management of cellular signaling. Protein kinases (PK) have significantly important roles in the cell signal transduction process. Protein kinases phosphorylate serine, threonine, tyrosine and histidine amino acids in a wide variety of molecular networks. Two main PK groups are distinguished; serine/threonine kinase and tyrosine kinases. MAPK (mitogen-activated protein kinases), ERK, EGFR (epidermal growth factor receptor), src, abl, FAK (focal adesion kinase), and JAK (janus family kinase) are considered as the main PK molecular networks. Protein kinases are closely related to the pathobiology of hematologic neoplastic disorders. For instance; JAKV617F point mutation-causing polycythemia vera and essential thrombocytosis occur at the position 617 in the JH2 domain of the JAK2 gene. The protein kinase inhibitor drugs targeting specific kinase molecules have already been developed and widely used in the field of Clinical Hematology. The existence of a local renin-angiotensin system (RAS) specific to the hematopoietic bone marrow (BM) microenvironment had been proposed two decades ago. Local BM RAS is important in hematopoietic stem cell biology and microenvironment. There are interactions among the local BM RAS and PK. For example, ACE2-ang(1-7)-Mas axis inhibits p38 MAPK/NF-КB signaling pathway. The Local BM RAS may have a role in the effect on PK in this biological spectrum. The aim of this review is to outline the functions of PKs in the pathobiology of hematologic neoplastic disorders.

Platelet Activation in Ocular Behçet's Patients with Posterior Segment Involvement.

PURPOSE: The aim of the present study was to assess platelet activation by analyzing three platelet activation parameters in ocular Behçet's disease (BD): mean platelet volume (MPV), platelet distribution width (PDW), and plateletcrit (PCT). METHODS: Twenty-nine patients with active ocular BD (Group 1), 40 patients with inactive ocular BD (Group 2), and 40 healthy adult individuals serving as controls (Group 3) were evaluated. All of the individuals had been performed the complete ophthalmologic evaluation. The levels of MPV, PDW, and PCT were measured in each group. RESULTS: The mean MPV level was 8.40 ± 0.97 in Group 1, 8.32 ± 1.04 in Group 2, and 7.77 ± 0.72 in Group 3. The mean PDW level was 15.12 ± 1.09 in Group 1, 14.97 ± 1.02 in Group 2, and 14.52 ± 0.82 in Group 3. The mean PCT level was 0.23 ± 0.07 in Group 1, 0.21 ± 0.04 in Group 2, and 0.18 ± 0.03 in Group 3. MPV, PDW, and PCT levels were significantly higher in ocular BD patients than controls (P < 0.05). CONCLUSION: Platelet activation may affect vascular occlusion in ocular Behçet's patients with posterior segment involvement. This result may be important in evaluating ocular BD patients.

Ankaferd hemostat (ABS) as a potential mucosal topical agent for the management of COVID-19 syndrome based on its PAR-1 inhibitory effect and oestrogen content.

COVID-19 due to the SARS-CoV-2 infection is a multi-systemic immune syndrome affecting mainly the lungs, oropharyngeal region, and other vascular endothelial beds. There are tremendous ongoing efforts for the aim of developing drugs against the COVID-19 syndrome-associated inflammation. However, currently no specific medicine is present for the absolute pharmacological cure of COVID-19 mucositis. The re-purposing/re-positioning of already existing drugs is a very important strategy for the management of ongoing pandemy since the development of a new drug needs decades. Apart from altering angiotensin signaling pathways, novel drug candidates for re-purposing comprise medications shall target COVID-19 pathobiology, including pharmaceutical formulations that antagonize proteinase-activated receptors (PARs), mainly PAR-1. Activation of the PAR-1, mediators and hormones impact on the hemostasis, endothelial activation, alveolar epithelial cells and mucosal inflammatory responses which are the essentials of the COVID-19 pathophysiology. In this context, Ankaferd hemostat (Ankaferd Blood Stopper, ABS) which is an already approved hemostatic agent affecting via vital erythroid aggregation and fibrinogen gamma could be a potential topical remedy for the mucosal management of COVID-19. ABS is a clinically safe and effective topical hemostatic agent of plant origin capable of exerting pleiotropic effects on the endothelial cells, angiogenesis, cell proliferation and vascular dynamics. ABS had been approved as a topically applied hemostatic agent for the management of post-surgical/dental bleedings and healing of infected inflammatory mucosal wounds. The anti-inflammatory and proteinase-activated receptor axis properties of ABS with a considerable amount of oestrogenic hormone presence highlight this unique topical hemostatic drug regarding the clinical re-positioning for COVID-19-associated mucositis. Topical ABS as a biological response modifier may lessen SARS-CoV-2 associated microthrombosis, endothelial dysfunction, oropharyngeal inflammation and mucosal lung damage. Moreover, PAR-1 inhibition ability of ABS might be helpful for reducing the initial virus propagation and mocasal spread of COVID-19.

In vitro analysis of the renin-angiotensin system and inflammatory gene transcripts in human bronchial epithelial cells after infection with severe acute respiratory syndrome coronavirus.

INTRODUCTION: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a recently identified coronavirus family member that triggers a respiratory disease similar to severe acute respiratory syndrome coronavirus (SARS-CoV). SARS-CoV and SARS-CoV-2 are very similar to each other in many respects, such as structure, genetics, and pathobiology. We hypothesized that coronaviruses could affect pulmonary tissues via integration with the critical immune genes after their interaction with renin-angiotensin system (RAS) elements. The aim of the present bioinformatics study was to assess expression changes of the RAS and non-RAS genes, particularly immune response genes, in the lung epithelial cells after infection with SARS-CoV. METHODS: Linear regression, hierarchical clustering, pathway analysis, and network analysis were performed using the E-GEOD-17400 data set. RESULTS: The whole-genome expression data of the lung epithelial cells infected with SARS-CoV for 12, 24, and 48 hours were analyzed, and a total of 15 RAS family and 29 immune genes were found to be highly correlated with the exposure time to the virus in the studied groups. CONCLUSION: RAS genes are important at the initiation of the infections caused by coronavirus family members and may have a strong relationship with the exchange of immune genes in due course following the infection.

Comparison of bortezomibcyclophosphamide- dexamethasone versus bortezomib-dexamethasone based regimens in newly diagnosed multiple myeloma patients.

The treatment landscape and clinical outcome of multiple myeloma (MM) patients have changed in the last decades, with an improved median survival of 8-10 years. This study aimed to evaluate the bortezomib, cyclophosphamide and dexamethasone (VCD) regimen versus bortezomib and dexamethasone (VD) regimen in patients with newly diagnosed MM. This study has been performed in a retrospective manner. One hundred and three patients with newly diagnosed MM who received chemotherapy at our tertiary care center between the years of 2009 and 2018 were evaluated. A total of 103 patients were included. The 5-year overall survival (OS) for patients who received VD regimen and patients who received VCD regimen were 75% and 83%, respectively. The OS for VD patients was 113.1±12.5 versus 122.2±9.5 months for VCD patients with no statistically significant difference (P=0.47). The 5- year PFS (progression free survival) for patients who received VD regimen and patients who received VCD regimen were 66% and 75%, respectively. The PFS for VCD patients was higher than the PFS for VD patients (67.1±7.4 versus 97.7±13.4 months), but no statistically significant difference was observed (P=0.59). Relapse rate (P=0.002) and mortality rate (P=0.01) were higher in VD group than VCD group and they were statistically significant. The OS and PFS were clinically longer in patients receiving VCD regimen than in patients receiving VD regimen, although not statistically significant. Cyclophosphamide should be given to patients at physician discretion and depending on patient's frailty function.

Pharmacobiological management of hemostasis within clinical backgrounds via Ankaferd hemostat (Ankaferd blood stopper).

Ankaferd hemostat (Ankaferd blood stopper [ABS], Istanbul, Turkey) is a hemostatic agent affecting red blood cell-fibrinogen interactions. ABS has been traditionally used in Anatolia as a hemostatic agent for centuries. ABS contains a standardized combination of the plants namely Glycyrrhiza glabra, Thymus vulgaris, Alpinia officinarum, Vitis vinifera, and Urtica dioica. The hemostatic effect of ABS depends upon the quick promotion of a protein network, particularly fibrinogen gamma, in relation to the erythrocyte aggregation. The aim of this review is to indicate pharmacobiological basis and clinical backgrounds of ABS. Current perspective for using ABS is to provide hemostasis and accelerating wound healing particularly in cases which are difficult to manage. Future controlled trials are needed to elucidate the actions of ABS with in hemostasis, antithrombotic, anti-inflammatory, anti-infective, antifungal, and anti-oxidative effects.

The Impact of Early Versus Late Platelet and Neutrophil Recovery after Induction Chemotherapy on Survival Outcomes of Patients with Acute Myeloid Leukemia

Objective: The prognosis of patients with acute myeloid leukemia (AML) is affected by factors that are both patient- and disease-specific. The aim of this study is to evaluate the impact of early versus late platelet and neutrophil recovery after induction chemotherapy on survival outcomes of AML patients. Materials and Methods: A total of 181 patients with AML who were treated in our tertiary center between 2001 and 2018 were evaluated. Neutrophil and platelet recovery times were accepted as the periods from the beginning of induction chemotherapy to a neutrophil count of ≥0.5x109/L and a platelet count of ≥20x109/L 3 days in a row, respectively. The median time to platelet recovery was 25 days (range=12-52) for all patients. Therefore, platelet recovery in the first 25 days was defined as early platelet recovery (EPR) and at ≥26 days it was defined as late platelet recovery (LPR). The median time to neutrophil recovery was 28 days (range=13-51) for all patients. Therefore, neutrophil recovery in the first 28 days was defined as early neutrophil recovery, and at ≥29 days it was defined as late neutrophil recovery. Results: The 5-year overall survival (OS) rates for patients who had EPR and LPR after induction chemotherapy were 62% and 23%, respectively (p<0.001). The 5-year disease-free survival (DFS) rates for patients who had EPR and LPR after induction chemotherapy were 57% and 15%, respectively (p<0.001). Conclusion: Short bone marrow recovery time may indicate better healthy hematopoiesis and marrow capacity associated with longer OS and DFS.

Evaluation of the Survival Outcomes of Multiple Myeloma Patients According to Their Plasmacytoma Presentation at Diagnosis

Objective: Multiple myeloma (MM) associated with extramedullary (EM) plasmacytoma has a poor therapeutic response and poor outcomes when treated with conventional chemotherapy. EM plasmacytoma is divided into two groups: the first group comprises tumors that are extending directly from osteolytic bone lesions (EM-B, bone-related), while the second results from plasmacytoma infiltration into soft tissues with no relationship to the bone (EM-S, soft tissue-related). This study aimed to compare the general characteristics and survival outcomes of transplant-eligible MM patients who had EM-S or EM-B and MM patients who did not have plasmacytoma at the time of diagnosis. Materials and Methods: This study was performed in a retrospective manner. The MM patients who were treated at our tertiary care center between January 2003 and January 2017 were evaluated retrospectively for the presence of plasmacytoma at diagnosis. Results: There were 141 (78.3%) MM patients who did not have plasmacytoma, 22 (12.2%) MM patients who had EM-B, and 17 (9.4%) MM patients who had EM-S at the time of diagnosis in this study. The 5-year overall survival was 63% in patients who had bone EM-B, 63% in patients who had EM-S, and 80% in patients who did not have plasmacytoma (p=0.02). The 5-year disease-free survival was 47% in patients who had EM-B, 35% in patients who had EM-S, and 54% in MM patients who did not have plasmacytoma (p=0.15). Conclusion: These findings lead us to suggest that MM patients with EM plasmacytoma at the time of diagnosis have poorer prognosis than patients without plasmacytoma, even if autologous stem cell transplantation is performed. The presence of EM involvement negatively affects survival outcomes.

WHO 2016 Definition of Chronic Myeloid Leukemia and Tyrosine Kinase Inhibitors

Philadelphia (Ph*)/BCR-ABL1-positive chronic myeloid leukemia (CML) is considered as a chronic life-long disease, which could be manageable with tyrosine kinase inhibitor (TKI) drugs. The aim of TKI drug treatment is to provide age- and sex-matched duration of life in a given patient with CML. Personalized CML treatment with TKI drugs is the key strategy. Individual treatment approach includes the harmonization of CML disease characteristics, clinical experience, and best available clinical evidence. Specific CML disease characteristics in a given patient include; CML disease risk, comorbidities, molecular profile, compliance, lifestyle, and drug off-target risk profile. CML research evidence includes; randomized clinical trials indicating the data on the efficacy, safety, tolerability, toxicity, possible long-term adverse events, and pharmacoeconomy of TKIs. Clinical and physician experience includes TKI availability, TKI reimbursability, drug experience, adherence, and BCR-ABL1 monitorization facilities. The key decision of choosing a TKI of choosing TKIs for CML should be made via the consideration of these variables. The aim of this paper is to outline the latest 2016 World Health Organization definition of CML and its proper management with TKI-class drugs.

The comparison of bone marrow kinetics between patients with acute myeloid leukemia and acute promyelocytic leukemia after induction chemotherapy.

Background and Aim: Recently, acute promyelocytic leukemia (APL) has shifted from the most hazardous to the best curable type of acute myeloid leukemia. Anthracyclines, all-trans retinoic acid (ATRA) and arsenic derivatives are the most important developments for the treatment of APL. ATRA promotes the terminal differentiation of malignant promyelocytes to mature neutrophils. We aimed to compare platelet and neutrophil recovery time after induction chemotherapy in patients with acute myeloid leukemia (AML) and APL.Materials and Methods: Two hundred and fifteen patients with AML and APL, who were diagnosed and treated in our tertiary care center between the years of 2001 and 2018 were evaluated.Results: One hundred and eighty one AML patients (84.2%) and 34 (15.8%) APL patients were included in this study. The time between neutrophil nadir after induction chemotherapy and neutrophil recovery was longer in APL patients than in AML patients [30.5 (4-52) vs. 20 (5-58), p < 0.001]. The time between platelet nadir after induction chemotherapy and platelet recovery was longer in APL patients than in AML patients [21.5 (4-42) vs. 17 (4-45), p = 0.02].Conclusion: Neutrophil and platelet recovery times were longer in APL patients than in AML patients in our present study. In 60 days, mortality rate was higher in APL patients than AML patients. Non-relapse mortality (NRM) rate was similar between two groups. There was a significant difference between two groups in terms of NRM causes. Platelet and neutrophil recovery time is very important because infection is the most important cause of NRM.

Indications and Outcomes of Splenectomy for Hematological Disorders.

BACKGROUND AND AIM: Splenectomy is a frequent component of the diagnosis and treatment of hematological disorders. The aim of this study was to define the indications and outcomes of splenectomy for benign and malign hematological disorders. MATERIALS AND METHODS: One hundred and two patients with hematological disease who had splenectomy at Hacettepe University Hospital between the years of 2010 and 2018 were evaluated. RESULTS: A total of one hundred and two patients were included in this study. The median age was 52 (20-82) years at the time of splenectomy. Most of the patients were female (57.9%). The median follow up time was 11.0 (0.03-87.9) months after splenectomy. Splenectomy was performed to diagnose thirty patients (29.4%). Seventy-two patients underwent splenectomy for the treatment of hematological disease (70.6%). Twenty-seven patients (90%) were diagnosed with various lymphomas. Two patients (6.7%) were diagnosed with hairy cell leukemia and one patient (3.3%) was diagnosed with large granular lymphocytic leukemia. CONCLUSION: In conclusion, an improvement in medical therapy, especially with monoclonal antibodies, the indications and outcomes of splenectomy for hematologic disorders have changed extremely in last years. Nevertheless, splenectomy has an important role for diagnosis and treatment of benign and malign hematological disorders.

Analysis of the Antiproliferative Effect of Ankaferd Hemostat on Caco-2 Colon Cancer Cells via LC/MS Shotgun Proteomics Approach.

Ankaferd hemostat (ABS), a traditional herbal extract, is a hemostatic agent used for wound healing and bleeding treatment. A standardized form of plants contains many biomolecules. In recent years, previous studies have demonstrated the antineoplastic effect of ABS. In the present work, we focused on the mechanism of its antineoplastic effect over Caco-2 colon cancer cells. The LC/MS-based proteomics method was used to understand the effect of ABS at the protein level. The results were evaluated with gene ontology, protein interaction, and pathway analysis. As shown by our results, ABS altered glucose, fatty acids, and protein metabolism. Moreover, ABS affects the cell cycle machinery. Moreover, we found that ABS induced critical cancer target and suppressor proteins such as carboxyl-terminal hydrolase 1, 60S ribosomal protein L5, Tumor protein D52-like2, karyopherin alpha 2, and protein deglycase DJ-1. In conclusion, the proteomics results indicated that ABS affects various cancer targets and suppressor proteins. Moreover ABS has systematical effect on cell metabolism and cell cycle in Caco-2 cells, suggesting that it could be used as an antineoplastic agent.

The impact of At1r inhibition via losartan on the anti-leukaemic effects of doxorubicin in acute myeloid leukaemia.

INTRODUCTION: Bone marrow renin-angiotensin system(RAS) modulates acute myeloid leukaemia(AML).The aim of this study is to clarify the relationships between RAS and AML, and to show the effect of losartan and doxorubicin treatment in AML cell lines. METHODS: AML cell lines including CESS, HL-60, MO-1, P31/FUJ, GDM-1 and KASUMI-3 were used as models in this study. RESULTS: After treating the six AML cell lines with a combination of losartan and doxorubicin, they were divided into two groups based on their behaviour: one became more sensitive to drug treatment (Group A) and the other had no change observed in behaviour after drug treatment (Group B). In silico analyses showed that Group A is involved in cellular apoptosis, while Group B is involved in tumour angiogenesis further supporting the in vitro results. CONCLUSION: The combined treatment of the AML cell lines with losartan and doxorubicin resulted in an increase in sensitivity of some of the cell lines. Those leukaemic cells are modulated via the induction of apoptosis, whereas the other cells resistant to the drug treatment are closely related to tumour angiogenesis indicating that RAS-AT1R seems to be differently expressed in different leukaemic blast cells and tumour microenvironments. Pharmaco-biological actions of RAS inhibitors may be different in distinct leukaemic cells based on the pathological behaviour of AML genomic subtypes.

Effects of Ankaferd Hemostat on Helicobacter pylori strains and antibiotic resistance

Background/aim: Ankaferd hemostat (ABS; Ankaferd blood stopper, Istanbul, Turkey) is a folkloric medicinal plant extract. The aim of this study was to determine the effect of Ankaferd hemostat (ABS) on the fate of Helicobacter pylori strains. The study also aims to determine alterations in the antimicrobial resistance of three different H. pylori strains in response to ABS exposure. Materials and methods: H. pylori Strain 1 was obtained from the culture collection ATCC 43504 and passaged three times for viability. Strain 2 was isolated from a gastric ulcer patient and Strain 3 was isolated from a gastritis patient. 1% of ABS was added to all of the strains and antimicrobial susceptibility was observed on 30 and 60 min after application. Results: The efficacy of ABS solutions in achieving significant logarithmic reduction in foodborne pathogens of H. pylori was observed in this study. This study showed that ABS has antibacterial (Anti-H. pylori) effects. Conclusion: Our present study indicated, for the first time, that ABS could act against H. pylori. ABS is clinically used for the management of GI bleeding due to benign and malignant GI lesions. Thus, the possible anti-H. pylori effect of ABS shall expand the therapeutic spectrum of the drug in GI lesions in relation to H. pylori infection such as peptic ulser disease (PUD) and lymphoid tissue (MALT) lymphomagenesis.