Electron configuration of group 8 chemical elements.
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Manning is a Principal Scientist (Rank Grade 9) in the Coasts & Oceans Group at HR Wallingford (UK) and has over 23 years of scientific research experience (in both industry and academia) examining natural turbulent flow dynamics, fine-grained sediment transport processes, and assessing how these interact, (including both field studies and controlled laboratory flume simulations). Andrew also lectures in Coastal & Shelf Physical Oceanography at the University of Plymouth (UK). Internationally, Andrew has been appointed Visiting / Guest / Adjunct Professor at five Universities (Hull, UK; Delaware, USA; Florida, USA; Stanford, USA; TU Delft, Netherlands), and is a highly published and world-renowned scientist in the field of depositional sedimentary flocculation processes. Andrew has contributed to more than 100 peer-reviewed publications in marine science, of which more than 60 have been published in international scientific journals, plus over 180 articles in refereed international conference proceedings, and currently has an H-index of 24. He supervises graduates, postgraduates and doctoral students focusing on a range of research topics in marine science. 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Out of these 118 elements, 92 elements originated from natural sources and remaining 26 elements have been synthesized in laboratories [1, 2]. The last naturally occurring element to be discovered was Uranium in 1789 [1, 3]. Technetium was the first man-made element to be synthesized in the year 1937 [2]. Recently in the year 2016, four of the man-made elements were included in periodic table. The four newly added elements goes by the name nihonium (Nh), moscovium (Mc), tennessine (Ts), and oganesson (Og), respectively for element 113, 115, 117 and 118 [4].
Discovery of Ruthenium had many twist and turns. A polish Chemist Jedrzej Sniadecki (1768–1838) in 1808 was first to announce the discovery of an element which he named Vestium after an asteroid called Vesta [3]. However, none of the contemporary Chemists were able to confirm his discovery. Later he again reported discovery of element 44 while working on the platinum ores from South America and published his results but again none of the fellow chemist were able to confirm the element 44 [4]. Due to repeated failures of his claim, Sniadecki got depressed and dropped the idea of further research on this element [1, 5]. After 20 years, a Russian chemist, Gottfried W. Osann, claimed the discovery of element 44. His discovery had the same fate as that of Sniadecki as none of his fellow chemist could repeat his results [5].
At last in the year 1844, another Russian chemist Carl Ernst Claus [also known in Russian as Karl Karlovich Klaus (1796–1864)] tried his luck on discovery of element 44. He succeeded in it as he gave positive proof about the new element extracted from platinum ores obtained from the Ural Mountains in Russia [6]. Claus had suggested the name of newly discovered element as Ruthenium after the name Ruthenia which was the ancient name of Russia. Earlier Osann had also suggested the same name for the element 44 [2, 5]. Ruthenium with atomic number 44 was given the symbol Ru. It is included in group 8, period 5 and block d in modern periodic table and it is a member of the platinum group metals [5].
Like other platinum group metals, Ruthenium is also one of the rare metals in the earth’s crust. It is quite rare in that it is found as about 0.0004 parts per million of earth crust [6]. This fraction of abundance makes it sixth rarest metal in earth crust. As other platinum group metals, it is obtained from platinum ores [7]. For instance, it is also obtained by purification process of a mineral called osmiridium [5].
In the modern periodic table, group 8 consists of four chemical elements. These elements are Iron (Fe), Ruthenium (Ru), Osmium (Os) and Hassium (Hs) [7]. Ruthenium has atomic number of 44, that is, it contains 44 electrons distributed in atomic orbitals and its nucleus has 44 protons and 57 neutrons (Figure 1). Electron distribution in atomic or molecular orbitals is called electron configuration which for Ru and the other group 8 chemical elements is shown in Table 1. Except for Ru, the electron configuration of group 8 elements shows two electrons in their outer most shell; Ruthenium has only one electron in its outermost shell. This tendency is quite similar to its neighboring metals such as niobium (Nb), molybdenum (Mo) and rhodium (Rh) [8].
Schematic of the electron configuration and nucleus of an atom of Ruthenium.
Atomic number | Element | Electron configuration | Number of electrons per shell |
---|---|---|---|
26 | Iron (Fe) | 1s2 2s2 2p6 3s2 3p6 4s2 3d6 | 2,8,14,2 |
44 | Ruthenium (Ru) | 1s2 2s2 2p6 3s2 3p6 4s2 3d10 4p6 5s1 4d7 | 2,8,18,15,1 |
76 | Osmium (Os) | 1s2 2s2 2p6 3s2 3p6 4s2 3d10 4p6 5s2 4d10 5p6 6s2 4f14 5d6 | 2,8,18,32,14,2 |
108 | Hassium (Hs) | 1s2 2s2 2p6 3s2 3p6 4s2 3d10 4p6 5s2 4d10 5p6 6s2 4f14 5d10 6p6 7s2 5f14 6d6 | 2,8,18,32,32,14,2 |
Electron configuration of group 8 chemical elements.
Any atom having same number of protons, but different number of neutrons is termed as an Isotope. Isotopes can be differentiated on the basis of mass number as each isotope consists of different mass number which is being written on the right of the element name [1, 7]. Mass number indicates sum total of proton and neutron present in the nucleus of atom [9]. Ruthenium has many isotopes although only seven of them are stable. Apart from seven stable isotopes, 34 radioactive isotopes of Ruthenium are also found [8]. The most stable radioactive isotopes are 106Ru, 103Ru, 97Ru having a half-life of 373.59, 39.26, 2.9 days, respectively. Other characteristics of the main isotopes are listed in Table 2 [8].
Ruthenium (Ru), Rhodium (Rh), Palladium (Pd), Osmium (Os), Iridium (Ir) and Platinum (Pt) form the Platinum group metals. Some of the fundamental properties of platinum group metals are summarized in Table 3 [8]. Ruthenium is silvery whitish, lustrous hard metal with a shiny surface. At room temperature, Ru does not lose its luster because it is unreactive in that condition but shows paramagnetic behavior [7]. At the higher temperature of around 800°C, Ru reacts with oxygen and gets oxidized [11]. It also reacts with halogens at higher temperature. As far as dissolution is concerned, Ruthenium does not dissolve in most of the acid or mixture of acids such as aqua regia which is a mixture of hydrochloric acid and nitric acid [7, 10]. When it is reacted with alkali it forms ruthenate ion which leads to dissolution of Ruthenium in alkalies (Eq. 1) [6].
Main isotopes of Ruthenium | |||
---|---|---|---|
S. No. | Isotopes | Abundance | Half-life |
1 | 96Ru | 5.54% | Stable with 52 neutrons |
2 | 97Ru | Synthetic | 2.9 days |
3 | 98Ru | 1.87% | Stable with 54 neutrons |
4 | 99Ru | 12.76% | Stable with 55 neutrons |
5 | 100Ru | 12.60% | Stable with 56 neutrons |
6 | 101Ru | 17.06% | Stable with 57 neutrons |
7 | 102Ru | 31.55% | Stable with 58 neutrons |
8 | 103Ru | Synthetic | 39.26 days |
9 | 104Ru | 18.62% | Stable with 60 neutrons |
10 | 106Ru | Synthetic | 373.59 days |
Physical properties of platinum group elements.
Ru | Rh | Pd | Os | Ir | Pt | |
---|---|---|---|---|---|---|
Atomic number | 44 | 45 | 46 | 76 | 77 | 78 |
Atomic weight | 101.07 u ± 0.02 u | 102.9055 u ± 0.00002 u | 106.42 u ± 0.01 u | 190.23 u ± 0.03 u | 192.217 u ± 0.003 u | 195.084 u |
Electronic configuration | Kr 4d7 5 s1 | Kr 4d8 5 s1 | Kr 4d10 | Xe 4f14 5d6 6 s2 | Xe 4f14 5d7 6 s2 | Xe 4f14 5d9 6 s1 |
Density(g/cc) | 12.2 | 12.41 | 11.9 | 22.59 | 22.56 | 21.45 |
Melting point(°C) | 2334 | 1963 | 1555 | 3033 | 2447 | 1768 |
Boiling point(°C) | 4150 | 3697 | 2963 | 5027 | 4130 | 3825 |
Electronegativity | 2.2 | 2.28 | 2.2 | 2.2 | 2.2 | 2.28 |
Characteristics of main isotopes of ruthenium.
As noted above, Ruthenium undergoes oxidation reaction to form Ruthenium oxide [11]. When Ruthenium oxide undergoes further oxidation in the presence of sodium metaperiodate, Ruthenium tetraoxide (RuO4) is formed (Eq. 2), with properties somewhat similar to those of OsO4, in that both are strong oxidizing agents. However, RuO4 differs from OsO4 since it can easily oxidize diluted form of hydrochloric acid as well as ethanol at normal room temperature [12]. At temperatures above 100°C, RuO4 get reduced to its dioxide. RuO4 also has specific stain property which is utilized in electron microscopy to investigate organic polymer samples [11, 13].
At lower oxidation states such as +2 or +3, Ru does not undergo oxidation reaction. Ruthenium reacts with hydroxide ions to attain higher coordination number [13]. Ruthenium does not form oxoanion readily as seen with iron. Ruthenium attains +7 oxidation states when it reacts with cold and diluted potassium hydroxide to form potassium perruthenate [14]. Ruthenium can also attain same oxidation state when potassium ruthenate gets oxidize in the presence of chlorine gas [9].
Coordination complex is the process where a center molecule makes bond with surrounding atoms or ions which are also known as ligands. Ruthenium readily forms coordinate complexes with different derivatives. It reacts with pentaamines to form different coordination complex. Ruthenium reacts with pyridine derivatives to form tris (bipyridine) ruthenium (II) chloride (Eq. 3) [15]. Ruthenium also reacts with carbon containing compounds. Ruthenium forms Roper’s complex when trichloride form of Ruthenium reacts with carbon monoxide [10, 15]. Ruthenium makes hydride complex when Ruthenium trichloride is heated in presence of alcohol which then reacts with triphenylphosphine to form chlorohydridotris (triphenylphosphine) ruthenium (II) (Eq. 4) [10].
Ruthenium acts as a catalyst in many reactions. In the olefin metathesis, the carbene and alkylidene complex of Ruthenium act as a catalyst. In Fischer Tropsch reaction (Eq. 5), Ruthenium also acts as a catalyst [16]. Fischer Tropsch reaction is a reaction in which liquid hydrocarbons are formed as a product of reaction between hydrogen and carbon monoxide. Decomposition process of ammonia also employs Ru as catalyst [17]. Ru also catalyzes group of reactions called “borrowing hydrogen reactions”. Borrowing hydrogen reaction is a reaction where two atoms of hydrogen are transferred to the catalyst to covert alcohol to carbonyl. The same reaction occurs in the conversion of alcohol to alkenes [5, 17].
Ruthenium carbonyl complex catalyzes the conversion of primary alcohol to aldehydes and secondary alcohol to aldehydes and ketones in the presence of a co-oxidant N-methylmorpholine-N-oxide (NMO) [8]. Ruthenium acts as a unique catalyst in oxidation reaction because of its varying oxidation state that ranges from −2 to +8 [6].
In recent years, there has been remarkable growth and evaluation in the field of coordination and organometallic chemistry of Ru. Many publications have appeared recently on the formation of Ru-based complexes and their applications in such areas as medicine, catalysis, biology, nanoscience, redox and photoactive materials. These developments can be related to the fact that Ru has the unique ability to exist in multiple oxidation states. Examples of these complexes and various applications of Ru are reviewed in the following sections.
Ruthenium (II)-arene complex has a structure of three-legged piano stool with a metal at the center in a quasi-octahedral geometry which is occupied by byan arene complex. 2-(naphthylazo)phenolate ligands reacts with chloro-bridged (g6-p-cymene) ruthenium complex [{(g6-pcymene)RuCl}2(l-Cl)2] in methanol having molar ratio 1:1 at room temperature leads to formation of monomeric ruthenium(II) complexes. The formed complexes (Figure 2) show the solubility in polar solvents (dichloromethane and acetone) and are insoluble in non- polar solvents (aspentane and hexane). It is stable in air and shows diamagnetic nature with the +2 oxidation state [6, 10].
Structure of (p-cymene) ruthenium (II) 2-(naphthylazo)phenolate complexes.
Polypyridine are coordination complexes containing polypyridine ligands such as 2,2′-bipyridine, 1,10-phenanthroline and 2,2′,6′2″-terpyridine. Polypyridines are multi-denated ligands which are responsible for characteristics property of metal complex they formed. Some of complexes show the characteristics of absorption of light by a process called metal-to-ligands charge transfer (MLCT). This said property of metal complex is due to the change in substituent to the polypyridine moiety. Among the polypyridine ligands for ruthenium complexes the mostly studied complex is Tris (bipyridine) ruthenium (II) chloride (Figure 3). It is a red crystalline salt having a hexahydrate form. Tris (bipyridine) ruthenium (II) chloride salt is prepared when aqueous solution of ruthenium trichloride reacts with 2,2′-bipyridine in the presence of reducing agent hypo-phosphorus acid. In this reaction Ru(III) gets reduced to Ru(II) [18].
Structure of Tris (bipyridine) ruthenium (II) chloride.
Ruthenium has a wide variety of application in diverse fields. Few of the applications of Ruthenium are listed below.
Ruthenium finds application both in electronic industry and chemical industry. In electrical industry it is used in manufacturing of electronic chips [19]. Chemically it is used in the form of anodes for chlorine production in electrochemical cells [20]. Ruthenium is used as a hardener when it is mixed with other metals to form alloy. This characteristic of ruthenium is used in the preparation of jewelry of palladium [18, 20]. When Ruthenium forms alloy with titanium it improves its corrosion resistant property. Ruthenium alloys also find application in manufacturing of turbines of jet engines [17]. Fountain pen nibs also contain Ru tips. Ruthenium has also application in therapy. For instance 106 isotope of Ru has application in radiotherapy of malignant cells of eye [11]. RuO4 is used in criminal investigations as it reacts with any fat or fatty substance having sebaceous pigments to give black or brown coloration due to formation of ruthenium dioxide pigments [12].
Ruthenium complexes tend to absorb light rays of visible spectrum. This property of ruthenium finds application in manufacturing solar cells for production of solar energy. [16] Ruthenium vapor get deposited on the surface of substrate and has magneto-resistive property. This property of Ru is used in making a layer or film on hard disk drives [12].
Ruthenium is used for determination of calcitonin level in blood. This determination is helpful in diagnosis and treatment of diseases related to thyroid and parathyroid glands. In treatment of medullary thyroid carcinoma (MTC), determination of calcitonin level plays an important role. The process of determination of calcitonin level involves one step sandwich assay method. This method is carried out in two incubation steps. Each incubation process takes 9 min each. In first incubation, 50 micro-liters of sample of biotinylated monoclonal human calcitonin specific antibody and monoclonal human calcitonin specific antibody labeled with ruthenium complex are incubated. This incubation leads to formation of sandwich like complex where human calcitonin is carrying both biotinylated and ruthenylated complex. After the first step, second incubation step is done where streptavidin-coated microparticles is added. Streptavidin-coated microparticle makes complex with biotin. After the incubation step, measurement is done. For measurement, the mixture of incubation is aspirated into measuring cells and micro particles of mixture are magnetically attracted to the surface of electrode. After that the unbound particles are removed. Voltage is applied on to the electrode and induction of chemi-lumiscent emission is done and after that the response is studied with photomultiplier [12].
Folate is the main constituent of synthesis of DNA. It is also essential for formation of red blood cells. Deficiency of folate leads to megalobalstic anemia. Deficiency of folate is estimated by determination of folate level in erythrocytes as well as serum. Ruthenium plays an important role in Elecys folate RBC assay in estimating folate deficiency in RBC. The process involved in folate determination is competition principle. This process involves three steps incubation method. In first incubation step folate pretreatment reagent is added which leads to release of folate from its binding sites (erythrocytes). In the second incubation step, Ru-labeled folate binding protein is added which makes complex with the sample. In the third incubation step streptavidin bounded microparticles are added which get attached to unbound sites of ruthenium-labeled folate binding protein. The whole complex is bound to solid phase via streptavidin and biotin. For measurement, the mixture of incubation is aspirated into measuring cells and microparticles of mixture are magnetically attracted to the surface of electrode. After that the unbound particles are removed. Voltage is applied on to the electrode and induction of chemi-lumiscent emission is done and after that the response is studied with photomultiplier [12].
Ruthenium is also employed in detection of cyclosporine by Elecsys cyclosporine assay. Determination of cyclosporine is an important aspect for management of liver, kidney, heart lungs and bone marrow transplant patients receiving cyclosporine therapy [12].
History of medical science shows metals like gold has always been used for medicinal purpose. Though it is known that metals may have beneficial effect for health, but the exact mode of activity remains unknown. Ruthenium also has been applied in treatment [21].
Immunosuppressant: Immunosuppressant is drug used to suppress hyperactivity of body’s immune system. An immunosuppressant Cyclosporin A which has wide application in treatment of disease like anemia and psoriasis eczema has shown side effects such as nausea, renal diseases, and hypertension. To modify the action of Cyclosporin A, complex is made with Ru(III). Ruthenium cyclosporin complex gives a stable compound which results in an inhibitory effect on T lymphocyte proliferation [22].
Antimicrobial action: antimicrobial drugs are drugs that inhibit microbial growth in human body. Ruthenium complex has its effectiveness against wide range of parasitic diseases. Microbial strains which are exposed to a certain kind of antimicrobial therapy become resistant to that drug. The resistance develops because the microbes mutate themselves against the organic compound of the drug. But with the formation of complex with certain metals the effectiveness of the drug increases as the microbes are unable to deal with the metal part of the organometallic complex of drug. In case of Chloroquine, Plasmodium species develops résistance against it, whereas when Chloroquine is complexed with ruthenium, resistance does not develop [23].
Antibiotic action: antibiotics are drugs which are made from one particular microorganism and act on the other microorganism. Synthetic antibiotics are also nowadays made in laboratory. Antibiotic exhibit their action by entering the cell of microbes and targeting any vital biosynthetic pathway. Ruthenium has upper edge if it gets complexes with synthetic antibiotics. Ruthenium being a metal has better tendency to bind to the cellular component similar to Iron. When an organic moiety gets bind to a metal ion, at that time sharing or delocalization of cations between the two moieties occurs. The change in charges among the component of drug increases the permeability of cellular component in favor of drug. For example, Thiosemicarbazone shows a remarkable increase in its activity due to formation of complex of Ru [24].
Inhibitory effect on nitric oxides: nitric oxide is a cellular component which is produced by many cells. The main physiological role of nitric oxide is to produce vasodilation. Nitric oxide does this action my increasing cellular level of cyclic-guanosine 3′,5′-monophosphate (CGMP) which is a secondary messenger in the physiological system. Over production of nitric acid can cause many disorders associated with respiratory system such as tumor of respiratory system. It also causes severe hypotension on over production. It also causes gastric inflammatory disorders. Ruthenium has beneficial effect in treatment of over production of nitric oxides. When ruthenium is administered in complex form such as ruthenium poly amino carboxylates, excess nitric oxide present in blood binds to this complex readily and reduces ruthenium to form an unabsorbable complex there by inhibiting its unwanted effects [25].
Anti-carcinogenic activity: cancer or carcinoma is a stage where body cells undergo uncontrolled proliferation and having invasiveness and metastatic property. To treat carcinoma, drug therapy aims at inhibiting synthesis of cancerous protein as well as inhibiting DNA replication. In market there are drugs such as Cisplatin which uses platinum as anticancer agent. Though platinum has shown better results in treatment of cancer but in some cancers, platinum is unable to show positive results. This shortcoming of Platinum made way for use of Ruthenium as a new entrant in treatment of cancer. Ruthenium shows the ability to bind to the DNA and inhibits its replication as well as protein synthesis. Ruthenium has low aqueous solubility which was the only drawback of it. This drawback was countered by using dialkyl sulfoxide derivative of ruthenium. The mechanism of action of ruthenium as an anticancer agent is that it causes apoptosis of tumor cells by acting at DNA level. Apoptosis is a controlled destruction of cells [17, 18].
Radiation therapy: in cancer treatment radiotherapy has also been used. Radiation therapy becomes beneficial only when it is proximal to the cancerous cell. The agents used in radiation therapy are called radio sensitizers. To increase the proximity to cancerous cells radio sensitizers’ complexes with ruthenium are used as Ru has the affinity to bind to DNA easily [18, 19].
Photodynamic therapy: it is a therapy where chemicals and electromagnetic radiations are used. In this therapy chemicals are targeted on the cancerous cell, these chemicals become cytotoxic when they interact with electromagnetic radiation. In this therapy Ruthenium find its application as it increases the access of these chemicals to the cancerous cells [20, 21].
Action on cancerous mitochondria: mitochondria are the power house of any cell. This makes it a potential target for anticancer therapy. Ruthenium red is a type of ruthenium which is used to stain mitochondria. Mitochondrial surface has some calcium entity on it. When ruthenium red is added, it reacts with this calcium and stains the mitochondria. Ruthenium red also has tumor inhibiting activity. However, ruthenium red is not preferably used clinically as it has major side effects [20, 22].
Effect on metastasis: metastasis is the ability of cancerous cell to spread in the body by lymphatic or circulatory system. A tumor cell more than 1 mm in size requires additional blood supply to spread in the body. Formations of new blood vessels are called angiogenesis. Drugs which act as anti-metastasis many inhibit this action. Ruthenium complexes anti-metastatsis drug namely NAMI-A does the same action by binding to the mRNA and production of denatured protein which gets accumulated on the surface of tumor making a hard film and prevents any blood supply to the tumor cell. This action inhibits the metastasis. Ruthenium has additional benefit that it easily crosses any cell so the reach of the drug increases [23, 26].
Ruthenium with atomic number of 44 and symbol Ru was discovered by Russian chemist Karl Klaus (1796–1864). In earth’s crust, it is quite rare, found in parts per billion quantities, in ores containing some of the other platinum group metals. It is silvery whitish, lustrous hard metal with a shiny surface. The ability of Ru to exist in many oxidation states is an important property of this rare element which plays an important part in its applications. Ruthenium readily forms coordinate complexes and these complexes have their applications in diverse fields such as medicine, catalysis, biology, nanoscience, redox and photoactive materials. In biomedical fields Ru is used for diagnosis and treatment purpose. For example, Ru is used for determination of calcitonin level in blood which is helpful in diagnosis and treatment of diseases related to thyroid and parathyroid glands. Also, Ru plays an important role in Elecys folate RBC assay in estimating folate deficiency in RBC. Ruthenium cyclosporin complex gives a stable compound which results in an inhibitory effect on T lymphocyte proliferation which shows its immune-suppressant action. Ruthenium complex has its effectiveness against wide range of parasitic diseases. Ruthenium shows the ability to bind to the DNA and inhibits its replication as well as protein synthesis. This property helps in the treatment of cancer. This chapter gives a brief account of the various properties of Ru which are exploited for applications in the medical field. It is likely that in the coming years, further research will lead to even more useful applications of this miraculous element.
Non-Hodgkin lymphomas (NHL) are the most common type of lymphoma, accounting for roughly 85% of this category of lymphoid neoplasia. NHL are most commonly classified as B-cell, T-cell, and natural killer (NK) cell lymphomas, according to their cell of origin. Diffuse large B-cell lymphoma (DLBCL) is a heterogenous class of aggressive lymphoma and is considered as the most common subtype of NHL. Several genetic anomalies such as point mutations, numerical alterations, and, more rarely, translocations and gene amplifications play a role in the pathogenesis of this class of B-cell lymphoma and have been related to specific histological and immunophenotypic subtypes [1].
\nThe incidence of diffuse large B-cell lymphoma (DLBCL) is increasing with age, from 0.3 cases/100,000/year in those between the ages of 35 and 39 years and 26.6/100,000/year for the 80- to 84-year-old age group [2, 3]. Mean age of diagnosis is 65 years for DLBCL and between 20 and 30 years of age for primary mediastinal subtype [4].
\nThis type of lymphoma is more common in Caucasians than African and Asian populations, while the extranodal involvement is more frequent in European and the Far East population than the American patients. The sex ratio ranges from 1.5 to 3.5/1 in favor of men [5].
\nDLBCL originates in mature B cells in certain stages of differentiation. Various changes occur in the B cell toward its malignant transformation, which are induced by genetic mutations. During their ontogenesis, the B cells are passing in the secondary lymphoid tissues where the antigen-dependent activation occurs. The activation and amassment of B cells in the secondary lymphoid tissue lead to the creation of the germinal center, which is critically dependent on BCL-6. If certain genetic alterations occur during lymphocyte development, then the determinatives of the neoplastic changes are settled. The type of lymphoma is determined by the stage of maturation of the B cell and by the type of anomalies that are interfering with their development and differentiation [6]. More specifically, the upregulation of B-cell lymphoma-2 (Bcl-2) protein expression and the inactivation of BCL6 which in turn blocks the apoptosis have both been observed in DLBCL.
\nMoreover, the elevation of the nuclear factor kappa-light-chain-enhancer of activated B cells (NFkB) and the upregulation of the avian myelocytomatosis virus oncogene cellular homolog (c-Myc) expression, leading to an increase in B cell proliferation, have also been cited [7]. The signal pathways for the B-cell receptor (BCR) and the activation of NFkB and downregulation of the B-cell lymphoma-6 (Bcl-6) pathways have a special pathogenic significance in DLBCL [7].
\nDLBCL can originate from three types of cells and can be classified accordingly as germinal center DLBCL (GCB DLBCL), from the centroblasts of the germinal center; activated B-cell DLBCL (ABC-DLBCL), from the plasmablasts that are involved in the terminal differentiation of the B cells; and primary mediastinal large B-cell lymphoma (PMBCL), which derives from thymic B cells. This classification has a prognostic utility, as the activated B-cell subtype has an unfavorable evolution [8].
\nApart from the cell-of-origin classification, several other classifications for non-Hodgkin lymphomas have been suggested during the last half-century, each with its own advantages and drawbacks: Rappaport, Kiel, International Working Formulation (IWF), and the Revised European-American Lymphoma classification (REAL). The last classification that is currently in use has been offered by the World Health Organization (WHO) in 2001 and was last revised in 2016. According to the maturation stage in which the B cell is in, and to the type of anomalies that occur during differentiation and maturation, DLBCL presents several variants and subtypes.
\nThe World Health Organization classification of the lymphoid neoplasms has systemized DLBCL into several subtypes, each with its own morphological, clinical, and immunohistochemical particularities. The 2016 revision of the WHO classification brought several updates concerning diagnostic and prognostic factors for DLBCL, such as the acknowledgment of the prognostic role of double-hit DLBCL-NOS, which involves the double expression of MYC and BCL2, or the newly introduced designation of DLBCL EBV+ (NOS) which is replacing the old term of “DLBCL EBV+ of the elderly” (Table 1) [9].
\nChronic lymphocytic leukemia/small lymphocytic lymphoma | \n
Monoclonal B-cell lymphocytosis | \n
B-Cell prolymphocytic leukemia | \n
Splenic marginal zone lymphoma | \n
Hairy cell leukemia | \n
Splenic B-cell lymphoma/leukemia, unclassifiable | \n
Splenic diffuse red pulp small B-cell lymphoma | \n
Hairy cell leukemia variant | \n
Lymphoplasmacytic lymphoma | \n
Waldenstrom macroglobulinemia | \n
Monoclonal gammopathy of undetermined significance (MGUS), IgM | \n
m heavy-chain disease | \n
g heavy-chain disease | \n
a heavy-chain disease | \n
Monoclonal gammopathy of undetermined significance (MGUS), IgG/A | \n
Plasma cell myeloma | \n
Solitary plasmacytoma of bone | \n
Extraosseous plasmacytoma | \n
Monoclonal immunoglobulin deposition diseases | \n
Extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma) | \n
Nodal marginal zone lymphoma | \n
Pediatric nodal marginal zone lymphoma | \n
Follicular lymphoma | \n
In situ follicular neoplasia | \n
Duodenal-type follicular lymphoma | \n
Pediatric-type follicular lymphoma | \n
Large B-cell lymphoma with IRF4 rearrangement | \n
Primary cutaneous follicle center lymphoma | \n
Mantle cell lymphoma | \n
In situ mantle cell neoplasia | \n
Diffuse large B-cell lymphoma (DLBCL), NOS | \n
Germinal center B-cell type | \n
Activated B-cell type | \n
T-cell/histiocyte-rich large B-cell lymphoma | \n
Primary DLBCL of the central nervous system (CNS) | \n
Primary cutaneous DLBCL, leg type | \n
EBV1 DLBCL, NOS | \n
EBV1 mucocutaneous ulcer | \n
DLBCL associated with chronic inflammation | \n
Lymphomatoid granulomatosis | \n
Primary mediastinal (thymic) large B-cell lymphoma | \n
Intravascular large B-cell lymphoma | \n
ALK1 large B-cell lymphoma | \n
Plasmablastic lymphoma | \n
Primary effusion lymphoma | \n
HHV81 DLBCL, NOS | \n
Burkitt lymphoma | \n
Burkitt-like lymphoma with 11q aberration | \n
High-grade B-cell lymphoma, with MYC and BCL2 and/or BCL6 rearrangements | \n
High-grade B-cell lymphoma, NOS | \n
B-cell lymphoma, unclassifiable, with features intermediate between DLBCL and classical Hodgkin lymphoma | \n
2016 WHO classification of mature B-cell neoplasms [9].
Over time, several variants and subtypes of DLBCL have been identified, according to their clinical features, localization, and morphology, which are all included in the recent REAL and WHO classifications. Extranodal DLBCL are relatively rare, accounting for 1–5% of NHL [10]. Less than 40% of DLBCL cases are originating in extranodal sites, especially in the gastrointestinal tract but also the mediastinum, bones, central nervous system (CNS), testes, and breast [11].
\nRoughly 10% of the total number of DLBCL is represented by PMBCL, a subtype which originates in medullary thymic B cells and affects young women, around the age of 30 and 40 years. They clinically appear as large mediastinal tumors sometimes involving the lungs and pericardium, which can lead to local compression with superior vena cava syndrome and airway obstruction. PMBCL is positive for pan-B markers, CD23 and CD30, and is negative to CD15 and sIg. BCL6, CD10, and interferon regulatory factor 4 (IRF4) can sometimes be positive [12]. Several chromosomal abnormalities can be found, such as gains in chromosomal arm 9p and 2p corresponding to JAK2 and cREL loci [13, 14]. Because of the diagnosis confusions between PMBCL and DLBCL with secondary mediastinal involvement, the reports concerning the survival rates of the two pathologic entities have been similar [15].
\nMore recently the improvements in diagnosis of PMBCL have allowed to observe a higher survival rate of this type of lymphoma compared to DLBCL [16]. Some variants of PMBCL, which own morphologic, immunophenotypical, and molecular resemblance with the nodular sclerosis subtype of Hodgkin lymphoma, are included in the category of B-cell lymphoma, unclassifiable, with features intermediate between DLBCL and classical Hodgkin lymphoma [9, 10, 17].
\nOnly a limited number of cases (2–3%) of cerebral neoplasms are lymphomas, the most frequent of which is DLBCL. Primary DLBCL of CNS can be both germinal center and non-germinal center, while some genetic anomalies suggest the implication of the microenvironment as a pathogenic factor [18].
\nThe clinical manifestations usually range from neurological deficits to psychiatric symptoms, but ocular involvement, seizures, and symptoms of increased intracranial pressure can also be found [18].
\nPTL are usually DLBCL (>80%) but can also classified as follicular, plasmablastic, Burkitt, mantle cell, or plasmablastic lymphomas in a minority of cases. Usually this type of lymphoma is diagnosed in stages 1–2, while in advanced stages, it can also affect the central nervous system, the liver, the skin, the Waldeyer ring, and the lungs [19]. PTL displays a modified expression of adhesion molecules that can be correlated with extranodal involvement. In most cases (60–96%) the cell of origin analysis indicates the activated B-cell (ABC) pattern. Several mutations have been observed, such as MYD88 mutations in 70% of cases, rearrangements of forkhead box protein P1 (FOXP1), programmed death-ligand 1 (PDL 1), and class II MHC transactivator (CIITA), as well as inactivation mutations of beta-2 microglobulin (B2M) gene [10].
\nPrimary breast lymphoma is a rare form of breast cancer, with cell of origin studies usually indicating an activated B-cell pattern. The cytogenetic analysis shows multiple chromosomal anomalies, such as trisomies 3 and 18 and translocations of chromosome 18 involving IGH/MALT1 [20].
\nPBoL is a rare type of lymphoma that is confined to the bone tissue, preponderantly affecting the middle-aged male patients. It should be differentiated from primary medullar lymphoma which, unlike PBoL, does not display cortical involvement [21]. The germinal center subtype is more common, frequently displaying BCL2, BCL6, and MYC rearrangements [10].
\nDLBCL patients have a wide spectrum of clinical manifestations, a frequent nodal involvement, but also, in up to 40% of cases, extranodal symptoms (skin, digestive and central nervous system, etc.). All these clinical manifestations, together with the general signs and symptoms (such as fever, weight loss, sweating) usually indicate a more aggressive phenotype. On rare occasions, the diagnosis is incidentally established when the clinical examination detects enlarged lymph nodes with no other signs or symptoms. Medullary involvement is uncommon in the early phases of the disease, as it is only found in less than 30% of cases [22].
\nAs for the paraclinical investigations, usual blood tests consist in complete blood count, lactate dehydrogenase and uric acid determination, serology, osteomedular biopsy, and cytologic exam of the cerebrospinal fluid. Computed tomography (CT) scan or, even better, positron emission tomography–computed tomography (PET-CT) is considered mandatory to assess the extension of the disease.
\nPET-CT is of great diagnostic and prognostic importance, but it is also necessary for the post-chemotherapy assessment, such that in case of negative scans at the end of chemotherapy regimen, the overall prognosis is favorable [23, 24]. The drawbacks of this technique consist of the possibility of false-positive results due to reversible thymic hyperplasia, infections, and sarcoidosis or as a consequence of hematopoietic growth factor therapy [23].
\nThe search for an exclusively cytologic classification of DLBCL is mostly abandoned, due to the wide observer-dependent variations in morphologic analysis and nosologic framing. The cytologic exam usually differentiates between centroblastic, immunoblastic, anaplastic, and T-cell/histiocyte-rich B-cell lymphoma.
\nThe subtype of DLBCL is decided by the genetic anomalies that occur during B-cell differentiation and maturation process [6, 25].
\nEven though gene expression profiling has the best accuracy for the identification of the cell of origin in DLBCL, its numerous drawbacks—such as the diverse cellular regulating pathways, some financial limitations, etc.—make its widespread clinical use still impractical. Consequently, several surface molecules have been evaluated for the differentiation between GCB and ABC DLBCL, which prove to be consistent with gene expression profiling.
\nThe Hans algorithm has structured DLBCL in these two main subtypes, by analyzing three essential markers: multiple myeloma 1 protein (MUM1), CD10, and bcl-6 (Figure 1). The same classification can be achieved by using the germinal center B cell-expressed transcript 1 (GCET1), CD10, BCL-6, MUM1, and FOXP1 biomarkers (the Tally and Choi algorithms) with a slightly better accuracy (Figure 2). All these algorithms are important, as the cell of origin classification tends to display a prognostic role, due to the fact that DLBCL-GCB subtype usually has a favorable prognosis compared to non-GCB subtype. To further add to the complexity of this classification, GC and ABC DLBCL subtypes also express several particularities in cell marker expression, activation pathways, and outcomes [26].
\nHans algorithm for DLBCL subtyping.
Choi algorithm for DLBCL subtyping.
However, the detection of other cell marker expression has become a fundamental component both in establishing an accurate prognosis and in the development of an optimal therapeutic algorithm.
\nThe treatment protocol in DLBCL did not witness significant changes during the last two decades. The widespread adoption of rituximab as an important adjuvant to standard chemotherapy protocol in CD20+ cases was a notable exception, which provided significant improvement in disease-free survival, overall survival, and complete remission rates, with limited toxicity. However, no less than 20% of patients diagnosed with DLBCL exhibit relapse after the initial response to R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone) regimen, while more than 15% of the patients exhibit primary refractory disease [27]. All these factors required the search for alternative therapeutic regimens, such as R-CHOEP (rituximab, cyclophosphamide, doxorubicin, vincristine, etoposide, and prednisone), R-miniCHOP (adjusted short-term R-CHOP therapy), R-ACVBP (adriamycin-cytoxan-vindesine-bleomycin-prednisone-rituximab), etc., that are strictly codified by several therapeutic protocols, such as the 2015 ESMO guidelines [10].
\nThe choice for any of these regimens should be adapted to the specific clinical status of each patient, as the exact therapy depends on a number of factors, such as the stage of the disease, age, and biological status. However, despite numerous attempts, no clear recommendation has been published thus far.
\nThe association of radiation therapy to chemotherapy offered conflicting results. While some studies showed some benefits for the combined therapy, most of the authors did not find any real advantages, especially in the case of localized disease without bulky mass [28]. There were some suggestions of a combined therapeutic algorithm, involving four R-CHOP cycles followed by either local radiation therapy or two more cycles of R-CHOP, in the case of complete remission. Armitage et al. pleaded for mandatory radiation therapy after six cycles of R-CHOP in the case of bulky disease [29]. More recently MInT (MabThera International Trial) study recommended the same therapeutic algorithm for localized bulky disease [30].
\nIn the case of localized disease, the therapeutic protocol varies with the specific organ involvement. For primary DLBCL of the testis, R-CHOP should be associated with radiotherapy and methotrexate or intrathecal cytarabine, due to the potential risk of CNS involvement. In case of documented CNS involvement before the onset of therapy, then high-dose intravenous and intrathecal methotrexate is advised. Conversely, cytarabine should be used instead of methotrexate for lymphomatous meningitis [31]. R-CEOP can be advised for patients with an altered cardiac function.
\nIn the case of advanced stage DLBCL, the therapeutic options varied greatly since the early 1980s. The third-generation regimens, ProMACE/CytaBOM (cyclophosphamide, doxorubicin, etoposid, Cytosar, bleomycin, vincristine, methotrexate, prednisone) and M-BACOD (methotrexate, bleomycin, doxorubicin, cyclophosphamide, vincristine, and dexamethasone), showed no benefits for complete remission or disease-free survival when compared to CHOP. However, a lower toxicity for CHOP was observed [32].
\nThe R-ACVBP regimen, consisting of rituximab, doxorubicin, cyclophosphamide, vindesine, bleomycin, and prednisone, for induction of remission, and consolidation with methotrexate, doxorubicin, and cyclophosphamide, was tested by the GELA (Groupe d’Etude des Lymphomes de l’Adulte) study on 379 patients. This regimen showed some advantages over the R-CHOP-21 but at the price of significantly higher toxicity [33].
\nGiven these circumstances, there is a growing need for new therapeutic alternatives, fitted for the morphologic, immunohistochemical, and genetic profile of each patient selected for individualized treatment protocol.
\nThe role of immunophenotype variability for the therapeutic outcome has long been the cornerstone for DLBCL management strategy, largely because the treatment of lymphomas evolves toward new therapies (immunotherapy, targeted therapy), which is made possible by analyzing the biology and the signaling pathways of this disease [34, 35]. Furthermore, a growing number of biological agents are available, varying from interferon to rituximab or radiolabeled antibodies, but also the more recent acquisitions in targeted therapy, such as ibrutinib, acalabrutinib, and daratumumab [36, 37, 38].
\nDespite the numerous advances and better understanding of the clinical, immunophenotypic, and genetic characteristics of DLBCL, but also in face of several breakthroughs in its treatment, the prognosis of this type of NHL has witnessed only modest improvements. The search for new biomarkers and efficient therapeutic agents in the context of future individualized treatment will be crucial in our quest for improved results.
\nAll authors shared equal contribution to this chapter.
\nNone.
ABC DLBCL | activated B-cell DLBCL |
B2M | beta-2 microglobulin |
Bcl-2 | B-cell receptor 2 |
Bcl-6 | B-cell receptor 6 |
CIITA | class II MHC transactivator |
c-Myc | avian myelocytomatosis virus oncogene cellular homolog |
CNS | central nervous system |
DLBCL | diffuse large B-cell lymphomas |
FOXP1 | forkhead box protein P1 |
GCB DLBCL | germinal center DLBCL |
GCET1 | germinal center B cell-expressed transcript 1 |
IRF4 | interferon regulatory factor 4 |
MUM1 | multiple myeloma 1 protein |
NFkB | nuclear factor kappa-light-chain-enhancer of activated B cells |
NHL | non-Hodgkin lymphoma |
PDL 1 | programmed death-ligand 1 |
PET-CT | positron emission tomography-computed tomography |
PMBCL | primary mediastinal large B-cell lymphoma |
PTL | primary testicular lymphomas |
R-ACVBP | adriamycin-cytoxan-vindesine-bleomycin-prednisone-rituximab |
R-CHOP | rituximab, cyclophosphamide, doxorubicin, vincristine, prednisone |
R-CHOEP | rituximab, cyclophosphamide, doxorubicin, vincristine, etoposide, and prednisone |
WHO | World Health Organization |
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