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In this chapter, we report and discuss the diagnosis and management of idiosyncratic drug-induced, or drug-associated, severe neutropenia, and agranulocytosis (neutrophil count of

Isothermal nucleic acid amplification is a simple process that rapidly and efficiently accumulates nucleic acid sequences at constant temperature such as 37 and 42°C. Isothermal nucleic acid amplification approach offers several advantages over temperature circle methods (such as PCR) including rapid assay results, cost-effectiveness, and portability. Two detection approaches based on circular strand-displacement polymerization reaction (CSDPR) were presented in this chapter for sensitive and specific thalassemia gene detection. One is a lateral flow strip biosensor based on CSDPR for semi-quantitative detection of thalassemia DNA. The other is a spectrophotometric DNA detection approach based on CSDPR for quantitative detection of thalassemia DNA.

Metastatic tumor blood perfusion and interstitial fluid transport based on 3D microvasculature response to inhibitory effect of angiostatin are investigated. 3D blood flow, interstitial fluid transport, and transvascular flow are described by the extended Poiseuille’s, Darcy’s, and Starling’s law, respectively. The simulation results demonstrate that angiostatin has the capacity to regulate and inhibit the formation of new blood vessels and has an obvious impact on the morphology, growth rate, and the branches of microvascular network inside and outside the metastatic tumor. Heterogeneous blood perfusion, widespread interstitial hypertension, and low convection within the metastatic tumor have obviously improved under the inhibitory effect of angiostatin, which suits well with the experimental observations. They can also result in more efficient drug delivery and penetration into the metastatic tumor. The simulation results may provide beneficial information and theoretical models for clinical research of antiangiogenic therapy strategies.

Introductory Chapter: Thalassemia - An Overview

Antiphospholipid syndrome is characterised by arterial and venous thromboembolic events and pregnancy morbidity (mainly, recurrent foetal losses), in the presence of antiphospholipid antibodies. Diagnosis is based on the presence of at least one laboratory and at least one clinical manifestation of antiphospholipid syndrome. There are also so-called “non-criteria” clinical features, and thrombocytopenia is one of the most important among them. Thrombocytopenia has been reported with a prevalence between 30 and 46% among patients with antiphospholipid syndrome. The pathogenesis of thrombocytopenia related to antiphospholipid antibodies is not absolutely clear. Binding of antiphospholipid antibodies to platelets and the promotion of platelet activation and aggregation thus thrombus formation must be an important mechanism as well as the immune-mediated clearance of platelets. Thrombocytopenia in antiphospholipid syndrome is usually mild and does not require clinical intervention. The presence of thrombocytopenia in patients with antiphospholipid syndrome is typically associated not with haemorrhagic complications, rather it can trigger thrombotic events. Other causes of thrombocytopenia, such as TTP, SLE, MDS, and ITP should be excluded. As thrombocytopenia is usually mild and it predicts later thrombosis, patients may be given platelet aggregation inhibitors and/or anticoagulant therapy. Anti-thrombotic treatment should be stopped only in case of severe thrombocytopenia.

Liquid biopsies represent an innovative methodology for cancer diagnostics and disease monitoring. The analysis of circulating cell-free nucleic acids (CFNA) and circulating tumour cells (CTC) are rapidly being adopted for quantitative and qualitative characterisation of the tumour genome and as a mode of non-invasive therapeutic monitoring. Circulating cell-free DNA (cfDNA) and CTC are representative of the underlying mutational profile of a cancer whereas the evaluation of extracellular RNA (exRNA) can be utilised as a prognostic biomarker thus providing critical biological information both at the time of diagnosis and during disease evolution. In this chapter, we will review the emerging utility of CFNA and CTC as biomarkers of prognosis and for both mutational characterisation and monitoring disease progression, and how these have the potential to provide additional information as an adjunct to bone marrow biopsies and conventional disease markers in multiple myeloma (MM). Emerging data suggest that liquid biopsies might offer a potentially simple, non-invasive, repeatable analysis that can aid in diagnosis, prognostication and therapeutic decision making in MM, with particular applicability in subsets of patients where conventional markers of disease burden may be less informative.

Prevalence of hepatitis C virus (HCV) infection is relatively low in children. However, seroprevalence rates of 10–20% have been reported among children who received repeated transfusion. The development and the severity of liver fibrosis are strongly related to the extent of the liver iron overload and to the presence of chronic hepatitis C (CHC). In CHC, liver iron overload has been suggested as a negative prognostic factor exacerbating inflammation with subsequent progression of liver fibrosis and decrease in antiviral therapy effectiveness. CHC may be suspected based on medical history or accidentally discovered abnormal liver functions. Hepatitis C is diagnosed by positive serology for viral antibodies and confirmed by polymerase chain reaction (PCR) to detect virus RNA. The treatment of HCV infection in children was difficult due to the limitations of pegylated interferon-α and ribavirin. In 2017, FDA approved the first direct-acting antiviral agents (DAAs) for children including ledipasvir/sofosbuvir in the adult dose, 90/400 mg, to treat HCV in children and adolescents aged 12 years and older or weighing at least 35 kg. Similarly, giving half the adult fixed-dose of ledipasvir/sofosbuvir, 45/200 mg, to children aged 6–11 years is still under clinical trials with promising results.

Interferon (IFN) is an effective therapy for multiple disorders. An infrequently reported side effect is thrombotic microangiopathy (TMA): thrombotic thrombocytopenic purpura (TTP) and hemolytic uremic syndrome (HUS). We published the first comprehensive review analyzing this association with the following observations: (1) there was a higher incidence of IFN-induced TMA in myeloproliferative disorders (chronic myelogenous leukemia (CML)) than that in nonmalignant disorders (multiple sclerosis (MS), chronic hepatitis C virus infection (HCV)); (2) mean age at diagnosis was 47 years; (3) there was rare association with hairy cell leukemia (HCL), Sezary syndrome (one case each) and no cases reported for polycythemia vera (PV); (4) sex distribution was balanced (exception of higher prevalence in females for MS); (5) TMA was insidious in onset with long incubation periods (average treatment duration 40.4 months); (6) comparative analysis of mean time (months) to onset of TMA ensuing cumulative IFN exposure was: MS 68.6 vs. CML 35.5 vs. HCV 30.4; (7) confirmed TTP (low ADAMTS 13 levels) was associated with the presence of an inhibitor; (8) outcome analysis revealed complete remission in 27 (40%), persistent chronic kidney disease in 28 (42%) and fatality in 12 patients (18%); (9) corticosteroids, plasma exchange (PEX) and rituximab are effective therapies.

The expression of a functional antigen receptor is necessary for cell survival of normal B lymphocytes and most B-cell neoplasms alike. When the genetic modifications of the B-cell receptor locus fail to produce a functional antigen receptor or result in deleterious mutations of a previously expressed receptor, the affected B cell will undergo apoptosis. The three physiological mechanisms that generate the B-cell receptor, VDJ recombination, somatic hypermutation, and class switch recombination, can induce double-strand DNA breaks and can specifically contribute to lymphomagenesis. On the other hand, the B-cell receptor activation and signaling pathways, which provide strong survival and proliferation signals to normal B cells, can support the growth and evolution of malignant lymphocytes. As a result, an otherwise structurally normal B-cell receptor can behave, from the functional perspective, as a true oncogene. In this chapter, we provide an in-depth discussion of the most recently discovered recurrent mechanisms involving the B-cell receptor in lymphoma pathogenesis. The discussion is structured around two major topics: (1) the genetic mechanisms that create a functional antigen receptor and their errors leading to oncogenic events, and (2) the pathogenic activation of the B-cell receptor signaling cascade. Finally, we will briefly comment on novel emerging therapies targeting the B-cell receptor at different levels.

Acute leukemia is the most common childhood malignancy, accounting for almost 35% of all childhood cancers. Acute myeloid leukemia (AML) represents 15–20% of pediatric acute leukemia. Majority of AML cases appear de novo, however a minority of cases can present as a secondary malignancy. AML is a highly heterogeneous disease and its diagnosis involves a combination of diagnostic analyses including morphology, immunophenotyping, cytochemistry, and leukemic blasts derived from peripheral blood or bone marrow demonstrating cytogenic and molecular characteristics. Through the identification of recurrent genetic mutations, it has been made possible to refine individual prognosis and guide therapeutic management. The current survival rate of children with AML is approximately 70%. The standard therapeutic regimen is a combination of cytarabine- and anthracycline-based regimens with allogenic stem cell transplantation in appropriate patients. Relapse in pediatric patients suffering from AML occurs in approximately 30% of cases, whereas death occurs in 5–10% of patients as a result of disease complications or chemotherapeutic side effects. In understanding the genetic basis of AML, targeted therapies will have the ability to reduce treatment-related morbidity and mortality. Here, we provide a comprehensive review of AML, its biology, diagnosis and therapeutic management in pediatric patients.

Platelets were mainly associated with coagulation and hemostasis; however, other biological effects have been attributed to platelets, including angiogenesis, extracellular matrix synthesis, inflammation, and immune response. Dengue virus infection causes 200 million cases of severe flu-like illness annually, escalating to life-threatening hemorrhagic fever or shock syndrome. Some hypotheses are postulated for immunopathogenesis of dengue, including antibody enhancement theory, T-cell activation of cross-reactive memory, and original antigenic sin. All hypotheses, to some extent, induce an overproduction or a skewed profile of cytokine release, giving rise to the term cytokine storm/cytokine tsunami. Although thrombocytopenia is typical of both mild and severe diseases, the mechanism triggering platelet reduction is incompletely understood. In dengue, platelets are one of the major cell populations affected by direct and/or indirect mechanisms of infection. It is common to observe both thrombocytopenia and platelet dysfunction in dengue, both strongly related to the clinical outcome. Thus, platelets are frequently affected in dengue, either for alteration of their own functionality, for “silent transport” of virus, or as an anti-viral immune cell. In this way, we describe some of functional aspects of platelets on dengue, observing circulating mediators, intraplatelet proteins contents, morphology, activation markers, and ability to interact with dengue virus.

Genetics has an important role in the risk stratification and management of the patients with acute myeloid leukemia (AML). Molecular testing can’t take the place of cytogenetic testing results, but has complementary role to help refine prognosis of the disease, especially within specific AML subgroups. Molecular genetic analysis of CEBPA, NPM1, and FLT3 is already the standard of care in AML patients, and mutations in several additional genes are assuming increasing importance. The French-American-British (FAB) classification and the World Health Organization (WHO) classification are the most classifications for AML. The aim of this chapter is a review on the role of genetic analysis in new treatments of AML.

Thrombocytopenia is a common complication of both chronic liver disease and liver transplantation (LT). The mechanism of thrombocytopenia is multifactorial implicating not only sequestration in the spleen, or in the graft, reduction in the mean platelet survival but also decreased platelet production due to low synthesis of thrombopoietin (TPO) or direct toxicity to the bone marrow. Platelets play a dualistic role in liver transplant, both beneficial and detrimental. The beneficial role of platelets is due to platelet-derived serotonin that is involved in liver regeneration. During surgery for liver transplant, in addition to the preoperative causes for thrombocytopenia, we have other mechanisms that will contribute to further deterioration of the platelets count and function: hemodilution, immunological reactions, and sequestration in the newly transplanted graft. This might result in a life-threatening level of thrombocytes. The concern is when we should treat thrombocytopenia because despite life-saving benefits, transfusion has also been related to complications and platelet transfusion has been identified as an independent risk factor for postoperative complications. Risks related to platelet concentrate administration are allergic reactions, alloimmunization, bacterial sepsis, and transfusion-related acute lung injury (TRALI). Administration of platelets is not indicated if there is no bleeding or immediate bleeding risk. New emerging therapies like thrombopoietin-receptor agonist will furthermore limit the administration of blood products.

Small non-coding RNAs (sncRNAs) are small regulatory molecules, which play key roles in fine-tune of all cell functions. In late 1970s and early 1980s, it was first determined that non-coding RNAs contribute to the cellular regulatory processes. The kingdom of sncRNAs is very numerous and it is clear that functions of different members of this family is different from each other and may be involved in normal and pathologic processes in cell. Recently it was investigated that sncRNAs and long non-coding RNAs play roles in cellular differentiation, proliferation, metabolic processes, bioenergetic regulation, cell death and inter-cellular communications, etc. In embryos, non-coding RNAs control maternal-zygotic transition, the maintenance of pluripotency, the pattering of the body axes, the specification and differentiation of cell types and morphogenesis of organs. Development of hematologic malignancies in humans, their course and regulation of resistance and sensitivity of tumorous cells to therapy are under the control of sncRNAs.

Acute myeloid leukemia (AML) is an extremely heterogeneous and deadly hematological cancer. Cytogenetic abnormalities and genetic mutations, though well recognized and highly prognostic, do not fully capture the degree of heterogeneities manifested in AML clinically. Additionally, current treatment of AML still largely depends on chemotherapy and allogeneic stem cell transplantation, with few options for personalized and molecularly targeted therapies. Proteomics holds promise for unraveling biological heterogeneities in AML beyond the scope of cytogenetics and genomics. In recent years, proteomics has emerged as an important tool for discovering new diagnostic biomarkers, enabling more prognostic patient classifications, and identifying novel therapeutic targets. In this chapter, we review recent advances in proteomic studies of AML, including an overview of AML pathology, popular proteomic techniques, various applications of proteomics in AML from biomarker discovery to target identification, challenges and future directions in this field.

Sickle cell nephopathy (SCN) begins early in childhood from failure of urinary concentration (hyposthenuria), albuminuria to hyperfiltration, hematuria and progression to falling glomerular filtration to end-stage renal disease and increased mortality. Renal involvement is more severe in homozygous individuals (HbSS) than in compound heterozygous patients (HbSC). The pathogenesis of SCN is multifactorial from hypoxia, acidosis, hemolysis, ischemia-reperfusion injury and albuminuria. The clinical manifestations depend on whether the main pathology is tubular, glomerular or a mixture of both abnormalities. This chapter offers a critical review of the recent literature and will highlight the pathophysiology, epidemiology, clinical manifestations and management of sickle cell nephropathy with particular focus on the major advance in the early diagnosis. Learning points: For SCN, the onset of hyperfiltration and albuminuria in infants and childhood is an opportunity to intervene early. There is no diagnostic markertest capable of detecting the onset of these changes. Moreover there is no reliable therapeutic agent to prevent or halt early changes due to SCN. The development of a marker of renal impairment in SCD such as such as Cystatin C assay if validated may be appropriate for wider clinical application.

More recently, DLBCL has witnessed advances in the molecular profiling and treatment of patients with refractory and relapsed disease. DLBCL is biologically and clinically a heterogeneous disease. Despite its aggressive behavior, DLBCL is a potentially curable disease with overall survival of 94 and 55% in patients with low and high rIPI scores, respectively. The combination of anti-CD 20 monoclonal antibody rituximab and cyclophosphamide, doxorubicin, oncovin (vincristine) and prednisone (R-CHOP) chemotherapy every 3 weeks is the first line treatment. Radiotherapy is reserved for the patients with bulky disease who fail to achieve complete remission after first line treatment. CNS prophylaxis is reserved for the patients with high lactate dehydrogenase (LDH) levels and involvement of more than one extranodal sites and for the patients with involvement of selective extranodal sites like testes and orbits (the sanctuary sites). Patients who suffer relapse after first line treatment receive high-dose chemotherapy supported by autologous stem cell transplantation (HDC/ASCT). Variants of DLBCL like double-hit (presence of MYC and BCL2/BCL6) and triple-hit (presence of MYC, BCL2 and BCL6) lymphomas are treated differently and these patients have worse outcome. Several novel immunotherapeutic agents like checkpoint inhibitors and chimeric antigen receptor T cell (CART) are being investigated in randomized trials on patients with DLBCL.

Leukemia, a blood cancer originating in the bone marrow, presents as a heterogeneous disease with highly variable survival rates. Leukemia is classified into major types based on the rate of cancerous cell growth and cell lineage: chronic or acute and myeloid or lymphoid leukemia. Histological and cytological analysis of the peripheral blood and the bone marrow can classify these major leukemia categories. However, histological analyses of patient biopsies and cytological microscopic assessment of blood and bone marrow smears are insufficient to diagnose leukemia subtypes and to direct therapy. Hence, more expensive and time-consuming diagnostic tools routinely complement histological-cytological analysis during a patient’s diagnosis. To extract more accurate and detailed information from patient tissue samples, digital pathology is emerging as a powerful tool to enhance biopsy- and smear-based decisions. Furthermore, digital pathology methods integrated with advances in machine learning enable new diagnostic features from leukemia patients’ histological and cytological slides and optimize patient classification, thus providing a cheaper, more robust, and faster diagnostic tool than current standards. This review summarizes emerging approaches to automatically diagnose leukemia from morphological and cytological-histological analyses.

Sickle cell disease (SCD) is a structural and monogenetic genetic disorder due to a mutation that occurs in the globin β-chain, resulting in the formation of hemoglobin S (Hb S), a protein composed of two normal, and two β-type mutant chains. Estimates indicate that the prevalence among live births is 4.4% in the world. The difficulty in circulating the sickle cell, its interaction with endothelial cells, leukocytes, platelets, endothelial dysfunction, and the abnormal expression of adhesion molecules permeate the beginning of the blood vessel occlusion process as well as pathophysiological aspects of SCD. Among the secondary complications are the stroke, pulmonary hypertension, leg ulcer, renal disorders, and all complications associated with vascular dysfunction. Clinical and biochemical markers of disease severity can be used to predict risk, prevent complications, and increase the expectation and quality of life of the SCD population. The entire scenario generated by Hb S has implications for the health and social inclusion of patients, so the treatment of the person with SCD needs an approach focused on the prevention of these complications in an individualized way.

The circulatory system plays a vital role in the survival of an organism by supplying it with essential nutrients, signaling molecules and eliminating the waste or toxic products from the body. This flow is tightly regulated by various factors, procoagulants support the formation of hemostatic plugs to prevent the leakage or blood loss and anticoagulants prevent the formation of unwanted clots. Disruption or dysregulation of procoagulants and anticoagulants lead to clinical complexities. In this chapter defects in the coagulation system, hereditary, acquired coagulation disorders, their diagnosis and recent clinical modulators of the coagulation system are discussed.

In the last two decades, heparin was widely used as an anticoagulant. Besides numerous advantages of heparin, some patients with heparin administration suffer from a side effect, the so-called heparin-induced thrombocytopenia (HIT), which can result in thromboses such as deep vein thrombosis, pulmonary embolism, occlusion of a limb artery, acute myocardial infarct, stroke, and a systemic reaction or skin necrosis. The basic on HIT complication have been investigated and led to clinical insights. Recent studies provided detail mechanisms among binding partners in HIT; especially, it has been shown that not only heparin but also a subset of antibody induce thrombocytopenia. In this chapter, insights into both heparin- and antibody-induced thrombocytopenia will be discussed and the novel mechanism of the autoimmune HIT caused by a subset of antibodies will be introduced.

Introductory Chapter: Thrombocytopenia

Hypoxia-inducible factor (HIF) is the central master regulator of adaptation to decreased oxygen availability in both physiological and pathological conditions. In leukemia, HIF regulates tumor cell metabolic regulation, metastasis, and other tumor-adaptive survival. However, the regulatory role of HIF in different types of leukemia, including myeloid leukemia, has been unclear. In this chapter, the focus throughout is on the aspects of roles of HIF in the tumor mitochondria metabolic change that are relevant to the assessment and treatment of myeloid leukemia. The connection of HIF with metabolic modification and anaerobic metabolism, along with epigenetic modification, contribute to abnormal biological and clinical behavior of myeloid leukemia, including response to treatment. We have also explored the metabolic requirements of tumor cell proliferation in an attempt to understand why tumor cells escape hypoxia-induced cell growth inhibition. We believe that a better understanding of the mechanistic links between HIF-regulated cellular metabolism, growth control, and epigenetic modifications could be useful for the indication of pharmaceutical agents in myeloid leukemia.

Hemoglobinopathies are a group of red blood cell productivity disorders, including α-thalassemia, β-thalassemia, and sickle cell disease (SCD), that are autosomal recessive and very common in Mediterranean, Middle Eastern, East Asian, and African countries. Thalassemia presents with the following clinical signs and symptoms: fatigue, weakness, yellowish skin, facial bone deformities, and abdominal swelling. Genetic studies have successfully characterized the key variants and pathways involved in hemoglobin F (HbF) regulation, providing new therapeutic targets for HbF reactivation. According to the current literature, using lentivirus vector for gene therapy and genome-editing-based treatment strategies for β-thalassemia and SCD have been discussed and well documented. According to current studies, novel treatments are becoming more important for thalassemia patients, because the consequences of supportive treatments are not sufficient for patients and their families. Supportive treatment does not have a positive effect on the survival rate of β-thalassemia patients. New treatments are empowering to develop develop a gene therapy for β-thalassemia and include pharmacological or disruption of BCL11A erythroid enhancer by CRISPR-CAS9 technology in addition to zinc-finger or transcription activator-like effector nuclease, and attempts at repairing the defective β-globin gene in hematopoietic stem cells by genome editing. These approaches are needed to improve for being more successful; gene addition has the advantage of making use of a single product applicable to all cases of β-thalassemia.

Introductory Chapter: Hemodynamic Management. The Problem of Monitoring Choice

Active, progressive hydrocephalus in children leads to increase of intracranial pressure, dilatation of cerebral ventricles, and decrease of intracranial compliance. These changes lead to disorder of regulation of cerebral circulation and development of cerebral hypoperfusion resulting in the secondary brain damage. Ependymal disruption, periventricular edema, and compression of the periventricular capillaries can be developed. Ischemia of the white matter can be developed due to hypoperfusion. But it is reversible if treated early and adequately. Transcranial Doppler sonography enables to determine hemodynamic parameters of cerebral circulation in various physiological and pathophysiological conditions. As transcranial Doppler sonography has been regarded to be noninvasive and appropriate for bedside treatment, it can also be applied in children at any age. The goal of this chapter is to assess changes of cerebral circulation in children with hydrocephalus and application of data from scientific studies of intracranial dynamics in children with hydrocephalus in clinical practice. The work is also focused on evaluation of impact of intracranial factors on Doppler parameters of cerebral circulation, especially in neonates with hydrocephalus. The ambition of this chapter is to improve indication and timing of drainage procedure in children with hydrocephalus by application of the results and clinical experience in daily clinical practice.

Beta thalassemia is the most frequent hemoglobinopathy worldwide. In patients with beta thalassemia major (BTM), the consequence of long-term life-saving transfusions is iron overload in liver, heart and endocrine glands. Hypogonadotropic hypogonadism is the most frequent endocrine complication. Recent progresses in the treatment of BTM dramatically improved life expectancy and quality of life of these patients, making the concern for fertility and pregnancy to gain importance. Therefore, we performed a review of the available data regarding hypogonadism in female patients with BTM. We found that hypogonadotropic hypogonadism is still frequently found in female patients with BTM. Pituitary iron overload seems to be the main factor contributing to hypogonadism occurrence, although iron-related damage of the ovaries and the genital tract cannot be excluded. The increased oxidative stress observed in BTM patients was hypothesized as a contributor to pituitary-gonadal dysfunction. Hypogonadism has significant consequences on quality of life, final height, bone health and fertility of the patients. Estro-progestative administration is essential in order to minimize consequences, although the best treatment regimen should be carefully weighted in each patient. Although spontaneous fertility is reduced by the presence of hypogonadism, it seems that ovulation-induction treatment with gonadotropins is effective in achieving pregnancies in majority of patients.

Myeloid sarcomas are extramedullary myeloid masses with associated tissue damage. Myeloid sarcomas usually arise before, during or after diagnosis of acute leukemia, most often AML. Majority of the patients with myeloid sarcoma respond to upfront systemic chemotherapy and sometimes bone marrow transplant, but it is unclear which patients will benefit from which treatments. This is primarily due to the paucity of knowledge on myeloid sarcoma. At present, there are no prognostic biomarkers for myeloid sarcoma, which can help in risk stratification in patients with myeloid sarcoma. Several studies have suggested that myeloid sarcoma is more likely to occur with certain translocations such as CBF and MLL rearrangements. In addition, sequencing analysis has identified several mutations in genes such as FLT3, NPM1, EZH2, and KIT. Nevertheless, there is still lack of knowledge to understand why particular leukemia migrates to the skin and soft tissues and becomes refractory to systemic therapy.

Introductory Chapter: Hematology in Times of Precision and Innovation

Vitamin C (ascorbic acid) is an essential nutrient with a number of beneficial effects on the human body. Although the majority of mammals can synthesize their own Vitamin C, humans and a few other species, do not produce it and depend on dietary sources for their Vitamin C supply. Among its many effects on cell function and metabolism, Vitamin C has shown, in vitro, a powerful anticancer effect against a number of human tumor cell lines, including myeloid leukemia. There are many different mechanistic explanations for the anticancer/anti-leukemic effects of Vitamin C and the aim of the present review is to illustrate these mechanisms, showing the results of some preliminary in vitro investigations, and outlining their potential clinical relevance.

Seventy seven women with 17–19 years of age examined central hemodynamics and its wave structure at rest, with orthopedic and psychoemotional load in different phases of the ovarian cycle (OC). It was established that in the luteal phase of the OC in the lying position, the blood pressure was higher than in other phases. In orthoprost in the luteal phase, in comparison with other phases, the growth of cardiac rhythm wave strength in the range of 0.04–0.15 Hz is observed, their concentration and connection with oscillations of the shock volume of blood, and the leveling of differences in blood pressure levels. The involvement of spontaneous baro-reflex sensitivity of the studied states and reactions is discussed.

Bone marrow (BM) clots are routinely sampled in aspiration tests, and their sections are prepared for histological observation by hematoxylin and eosin (H&E) staining. However, H&E-stained sections are considered less informative than those stained by the May-Grünwald Giemsa (M-G) stain; thus, diagnosis using H&E-stained clot samples is challenging for pathologists. In fact, the diagnostic evaluation is limited to the observation of cellular morphology and the myeloid-erythroid cell ratio. Pathologists leave cellular observation to laboratory hematologists, who generally use M-G staining. In this chapter, the utility of bone marrow clot specimens for diagnosis by H&E staining is reviewed. Specifically, the review provides a descriptive and illustrative explanation of the diagnosis of acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), and myelodysplastic syndrome/myelocytic proliferative neoplasm (MDS/MPN) and demonstrates the possibility of diagnosis on the basis of the characteristic features of blast cells. Clot specimens appear to be useful for the diagnosis of hematopoietic dysplasia by pathologists, and this approach can provide more informative findings for hematologists.

Introductory Chapter: Myeloid Leukemia

In the human placenta, fetal blood circulates in the blood vessels of the villous tree while maternal one circulates in the intervillous space, the surroundings of the villous tree. Previously, the computational model of the villous tree, whose stem villi actively contract because of the contractile cells, has been developed. The result of the computation indicated that the displacement caused by the contraction would be helpful for the fetal and maternal circulations and can be combined with the other measurements for blood circulations in the placenta. Hypoxia in the placenta is classified into the following categories: preplacental hypoxia, uteroplacental hypoxia, and postplacental hypoxia. The number and the form of the terminal villi are altered by hypoxia. Assuming that increase in the terminal villi causes a higher shear elastic modulus of the placenta, this villous tree model is useful to estimate the influence of hypoxia on the blood circulations. In this chapter, how these three types of hypoxia influence the blood circulation in the placenta by the aforementioned computational model are discussed. While preplacental hypoxia and uteroplacental hypoxia would cause similar displacement in large regions, postplacental hypoxia would do vice versa. All the types might make the fetal and maternal blood circulations difficult.

Internal tandem duplication mutations in the FLT3 gene (FLT3/ITD) are detected in 10–15% of children and 30% of adult patients with AML and are associated with an extremely poor prognosis. Although several antagonists against FLT3/ITD have been developed, few of them are effective for the treatment of FLT3/ITD+ AML because of the emergence of drug-resistant cells. The mechanisms responsible for drug resistance include the acquisition of additional mutations in the FLT3 gene and/or activation of other prosurvival pathways such as microenvironment-mediated resistance. Recent studies have strongly suggested that the reciprocal interaction between the microenvironment and AML cells generates specific machinery that leads to chemoresistance. This chapter describes the molecular mechanism responsible for the refractory phenotype of FLT3/ITD+ AML cells resulting from the communication between the microenvironment and FLT3/ITD+ leukemia cells. Understanding this mechanism enables the discovery of novel and innovative therapeutic interventions for resistant FLT3/ITD+ AML.

Thalassemia is a significant health problem worldwide. There are two main classifications, α- and β-thalassemias, which are usually caused by the defective synthesis of the α-globin, and which are commonly caused by different mutations of the β-globin chain. Different hemoglobin mutations have been identified to date. Thalassemias can result in profound anemia from early life and, if not treated with regular blood transfusions, can lead to death in the first year. Prenatal diagnosis of thalassemia is the essential part of preventive medicine and is currently dependent on the use of invasive diagnostic tests within the first 2 months of pregnancy. These diagnostic techniques carry a small but significant risk of fetal loss up to 1%. Molecular diagnostic methods have been developed for genotyping thalassemias based on PCR techniques and high-throughput technologies. Noninvasive tests using cell-free DNA (cfDNA) from a maternal blood sample is also an alternative method, thus eliminating the risk of miscarriage. This chapter summarizes the current invasive approaches and the noninvasive methods using cell-free fetal DNA as new molecular diagnostic methods for genotypic diagnosis of thalassemia in clinical practice. Prevention strategies that encompass carrier screening, genetic counseling, and prenatal diagnosis are discussed.

This chapter focuses on anti-red blood cells antibody and anti-globulin test. The relationship between warm or cold antibody and hemolysis is explained. Direct anti-globulin test (DAT) is a useful clinical examination tool on the diagnosis of autoimmune hemolytic anemia (AIHA); however, false positive or negative results are sometimes detected. This chapter shows the disposition about the surroundings of the IgG antibody in DAT examination. In addition, this chapter contains pointing issue on the diagnosis of AIHA. Some pitfalls about diagnostic AIHA are presented from our experienced cases. To diagnose the background diseases such as lymphoproliferative disorders or autoimmune diseases under the analysis of secondary AIHA is important.

Acute myelogenous leukemia (AML) is a clonal, malignant disease of hematopoietic tissue that is characterized by accumulation of abnormal (leukemic) blast cells, principally in the bone marrow. Representation of these genetic mutations and the involvement patterns seems to follow specific and temporally ordered fluctuating manners. Somatic mutations in these genes are represented as a variety of recurrent chromosomal abnormalities, e.g., t (8;21), t(15;17), etc., or by the presence of prognostic markers, e.g., FLT3, MLL, NPM1 and CEBPA as well as encoding epigenetic modifiers, such as DNMT3A, ASXL1, TET2, IDH1, and IDH2, are commonly acquired early and are present in the founding clone. The same genes are frequently found to be mutated in elderly individuals along with clonal expansion of hematopoiesis that confers an increased risk for the development of hematologic cancers. Furthermore, such genomic changes may persist after therapy, lead to clonal expansion during hematologic remission, and eventually lead to relapsed disease. Majority of genetic data are now being used to classification, risk stratification, and clinical care of patients. The unprecedented molecular characterization provided by advanced and deeply sensitized molecular assays like next-generation sequencing (NGS) offers the potential for an individualized approach to treatment in AML, bringing us one step closer to personalized medicine.

Intracerebral hematoma occurs in about 35/100,000 population and the incidence is likely increase over the next few decades as the population ages. The most common causes are hypertension and amyloid angiopathy. Bleeds due to these two causes are classified as primary while all other causes, such as AVM bleeds, coagulopathies, and so on, are classified as secondary. Primary tissue damage due to the intracerebral hematoma is followed by edema, neuronal damage, and secondary damage due to cellular breakdown. Basal ganglia are the most common site of intracerebral hemorrhage, accounting for nearly 50% of cases. CT scan, CT angiogram, DSA, and MRI are the investigations of choice. The initial management is medical, with control of blood pressure and antiedema measures forming the mainstay of treatment. Surgical option includes external ventricular drainage, endoscopic evacuation of hematoma, craniotomy and evacuation of hematoma, and decompressive craniectomy and is usually reserved for patients who deteriorate while on treatment.

Oral tissues are constantly exposed to damage from the mechanical effort of eating and from the invasion of foreign microorganisms such as bacteria, fungi, and virus. In healthy oral tissues, there is a balance between symbiotic bacteria and cells from the innate immune system, mainly neutrophils. When this balance is broken, inflammation appears and more immune cells are recruited to the gingiva. Neutrophils form a barrier against dysbiotic bacteria. However, when neutrophils are insufficient, bacteria thrive causing periodontitis, a chronic inflammatory disease that destroys the tooth‐supporting tissues or periodontium. Damage of periodontal tissues leads to tooth loss, and in severe cases, it can also affect systemic health by increasing a person's risk for atherosclerosis, rheumatoid arthritis, diabetes, and even cancer. The mechanisms neutrophil employ to keep a balance with bacteria in order to maintain healthy oral tissues is the focus of this chapter. We discuss how neutrophil antimicrobial functions keep bacteria at check and how some dysbiotic bacteria block neutrophils to promote an inflammatory state. Also, novel therapeutic approaches for periodontitis are discussed.

Polymorphonuclear neutrophils (PMNs) are the most abundant leukocytes in the blood and are considered as the first line of innate immune defence against infectious diseases. However, PMN cells have a crucial function in both innate and adaptive immune responses. Neutrophils have several mechanisms to control pathogens, and one of them is their capability to form neutrophil extracellular traps (NETs) that may control infection. NETs have the capacity to trap microorganisms, kill them, or avoid their dissemination. The aim of this chapter is to provide a comprehensive review on NETs, the cells that produce them, and some of the mechanisms involved in their formation, their role in the immune response, and the pros and cons of NETs, focusing mainly on infectious diseases.

Neutrophils, as the main cells of the first line of host defense against microbial pathogens, are responsible for pathogen recognition, inhibition of pathogen spreading into the host tissue, and finally, killing the invader cells. Neutrophils carry out these functions via numerous mechanisms, including a relatively recently described activity based on a release of neutrophil extracellular traps (NETs), a process called netosis. NETs are structures composed of DNA backbone, decorated with antimicrobial factors, derived from neutrophil granules. The structure of NETs and their enzymatic and microbicidal inclusions enable efficient trapping and killing of microorganisms within the neutrophil extracellular space. However, the efficiency of NETs depends on neutrophil ability to recognize pathogen signals and to trigger rapid responses. In this chapter, we focus on possible pathways involved in the release of NETs and summarize the current knowledge on triggers of this process during bacterial, fungal, protozoan, and viral infections. We also consider the mechanisms used by microorganisms to evade NET-killing activity and analyze the harmful potential of NETs against the host cells and the contribution of NETs to noninfectious human diseases.

It is now known that neutrophils make up a population of complex cells with great plasticity, challenging the old view of neutrophil association with tissue damage and early phases of infection. Here, we discuss different contexts in which these cells can induce anti-inflammatory responses. Although distinct surface markers and cytokines profiles were shown, the most reliable characterization of suppressor neutrophil subtypes relies on their functional characteristics. One important example of inhibitory neutrophils generation comes from in vivo treatment with G-CSF, for 5 days, as for hematopoietic-stem-cell-transplantation (HSCT). In this case,donor blood is enriched in degranulated granulocytes harboring a functional regulatory phenotype, characterized by IL-10 production. These cells, when transferred together with HSCT, are able to reduce graft-versus-host-disease, being influenced by Treg cells and influencing them back. Importantly, this protection is long lasting and specific, keeping immunocompetence to other antigens. This regulation is paramount in HSCT, and represents a simple approach to be applied in humans. In summary, we discuss the interaction of neutrophils with other cell types and its consequence in immunomodulation. We believe these features confer an important bridge between innate and adaptive immune system, building a new knowledge for an underestimated cell type.

Introductory Chapter: An Introduction to Hypertension-Related Intracerebral Hematomas

Intracerebral hemorrhage (ICH), defined as bleeding within the brain parenchyma, remains a challenging and controversial neurosurgical entity to treat. ICH has a broad range of etiology—stemming from complications associated with traumatic head injury to complications of hemorrhagic stroke. The role of medical management lies in optimizing blood pressure and intracerebral pressure, preventing secondary injury from complications of the hematoma such as seizures, and correcting coagulopathy. Given the mass effect of a hematoma and the possibility of expansion, surgical interventions attempt to evacuate the clot to restore normal intracerebral pressure and prevent worsening neurologic injury. This chapter reviews the recent controversy associated with surgical evacuation of intracerebral hemorrhage placing particular emphasis on the size and location of the hemorrhage and the methods used to evacuate the expanding ICH. Moreover, this chapter reviews considerations and therapeutic goals of the preoperative and postoperative window to minimize complications and optimize patient care.

Polymorphonuclear (neutrophil) granulocytes (PMNs) are an essential part of the innate immune responses and key instigators and effectors of the underlying pathological mechanisms (endothelial damage, interstitial histolysis, cytokine production, phagocytosis) leading to post-injury inflammation and secondary tissue injury. In 2004, the formation of neutrophil extracellular traps (NETs) was identified as an additional defence mechanism of PMN against microbes. The understanding of complex regulation of neutrophil functions and NET formation is essential for differentiating between healthy and pathological inflammatory response, which frequently determines if patient recovers uneventfully or develops catastrophic complications. Recent discoveries have revealed the potential role of NETs in the pathogenesis of a wide range of non-infectious diseases, including post-injury sterile inflammation. In such conditions, both spontaneous NET formation and impaired NETosis are documented. In this chapter, we review the evidence for the role of NETs in post-injury inflammation, the key molecular and cellular participants in pathological NET formation, the clinical relevance of NETs in post-injury complications and the therapeutic potential of NET inhibition/clearance.

Objective:The objective is to study the relationship between cell age and function in the process of the red blood cell (RBC) normal metabolism and investigate the functional changes of red blood cells in the preservation process.

Acute upper gastrointestinal bleeding (UGIB) is a potentially lethal and frequent digestive disease. It is mainly divided into the nonvariceal UGIB and variceal bleeding according to the source of bleeding. Red blood cell transfusion is the core therapeutic option for the management of acute UGIB. In this chapter, we reviewed the primary evidence from meta‐analyses and large‐scale randomized controlled trials regarding red blood cell transfusion strategy for acute UGIB.

Platelets are classically used in the clinic to maintain hemostasis, while recent evidence has shown the important role for platelets in the host inflammatory and immune responses. In fact, platelets in vivo produce different mediators such as cytokines or chemokines, which may be involved in the course of disease treatment, thus platelets transfusion is often an effective therapy in many cases. It is well known that platelets can regulate neutrophils, lymphocytes and other immune cells behavior in immune response, thus directing these immune cells onto the damaged tissues, organs or infected sites. On the other hand, platelets can induce neutrophil extracellular traps release in response to bacterial or viral infection. All the characterized novel profile of platelet, if not all, at least in some situations, should be take into consideration when platelets have to be transfused into patients.