Duffy blood group system phenotypes and prevalence. Reproduced with permission and modification.
\r\n\tAn important part of the book will consider electrodes (materials, configurations, contacts with biological matter) as responsible tools for the acquisition of bioimpedance data correctly. Implementations in wearable and implantable health monitors are the proposed book topics. Detecting of different pathogens by the aid of lab-on-chip (LoC) devices for point-of-care (PoC) and need-of-care (NoC) diagnostics is expected. Also, express analysis of biological matter (blood and other body fluids) is included. Electronics connected to electrodes for receiving the bioimpedance signals for further processing belongs to sensing techniques and will be considered.
\r\n\tDevelopment and application of software tools for information extracting from the acquired bioimpedance data, automatic identification of bioparticles and the decision making for diagnosing and treatment are very welcome chapters in the present book.
Infective endocarditis (IE) is characterised by a microbial infection that involves the endocardial surface of the heart; it most often denotes infection of the heart valves or an intracardiac device [1]. Less commonly it involves septal defects, mural endocardium and the subvalvular apparatus [2]. The classic lesion is a vegetation, which is composed of platelets, fibrin enmeshed with microorganisms and inflammatory cells [3]. Infective endocarditis is caused by many different species of bacteria such as staphylococci, streptococci, enterococci and slow-growing Gram-negative coccobacilli.
In the 1950s IE secondary to intravenous drug use was first described [4]. Right-sided infective endocarditis (RSIE) secondary to intravenous drug use is a distinct entity and will be reviewed in this chapter.
Intravenous drug abuse (IVDA) is a recognised risk factor for IE. Intravenous drug users are at a seven times higher risk for infective endocarditis compared to patients with rheumatic heart disease or prosthetic valves [3].
Over the last decade there has been a steady increase in number of cases related to IE due to intravenous drug use. Between 2000 and 2008 the rate of IE due to IVDA has increased from 6 to 8% hospitalisation to 12% in the year 2013. During this period there has been an increase in IVDA related IE cases amongst younger white population. A similar distribution was noted between males and females [5].
In United States, North Carolina, a study reported a 12-fold increase in hospitalisations for intravenous drug use related IE over the last decade [6].
In the past IE related to IVDA was predominantly disease of men. A recent study has shown that there is a general increase in the rate of IVDA associated IE in the United States, with a relatively higher proportion of women compared to previous studies [5, 6]. In a recent South African Study, Meel et al. reported an increase in the incidence of IE related to IVDA amongst Africans. These were predominantly male and majority were HIV infected [7].
Infective endocarditis involving the right side accounts for 5–10% of cases of IE [8, 9].
RSIE may occur in patients with intracardiac devices but in intravenous drug users it is usually associated with HIV infection [9].
HIV infection in intravenous drug users is associated with a higher rate of IE compared to HIV uninfected users [10, 11]. Further, immunosuppression with lower CD4 count is associated with a higher predisposition to IE [9].
Intravenous drug use related IE involves the tricuspid valve in 46–78% of the cases, mitral valve in 24–32% of cases and the aortic valve in 8–19%. About 16% of the patients have multiple valve involvement. In the majority the infection occurs on the native valves. Intravenous drug use is characterised by recurrent infective endocarditis of the native valves [3].
The most common organism isolated in IVDA related IE is Staphylococcus aureus. It accounts for greater than 50% of the organisms cultured [3]. It tends to commonly infect the native tricuspid valve. In contrast streptococci and enterococci infect damaged valves, mostly aortic and the mitral valve. Other organisms include fungi, Pseudomonas aeruginosa and Gram-negative bacilli. Injection of contaminated material predisposes drug addicts to less commonly encountered organisms such as Corynebacterium species, Lactobacillus, Neisseria species and Bacillus cereus. In 3–5% of cases Polymicrobial infection is present [3].
The tricuspid valve is the most commonly involved valve in RSIE due to IVDA. Injection of recreational drugs results in entry of particulate matter such as talc into the circulatory system resulting in structural damage to the endothelium of the valve [12, 13]. Similarly, the left-sided valves get damaged by particulate matter that is less than 10 mm in size and is able to cross the pulmonary circulation [14]. The use of cocaine is associated with greater frequency of IE in IVDA. The possible mechanisms postulated include the ability of cocaine to cause vasospasm and tissue damage to the myocardium. It is also procoagulant and thus can cause thrombus formation and thus producing a nidus for bacterial seeding the damaged valve tissue [8]. Further, it has been postulated that intravenous drugs can result in pulmonary hypertension leading to increased turbulent blood flow across the valve resulting in endothelial damage to the right-sided heart valves.
The pathogenesis of formation of vegetation is complex. It involves interaction between the host, the organism, the endothelium, hemostatic pathways, the ability of the hosts immune system to eliminate the organism and the virulence of the specific microorganism [3].
The microorganism once in the blood stream tend to attach themselves to the valve surface and proliferate at sites of endothelial damage resulting in further damage to the valve tissue. The microorganisms initially attach to the platelet-fibrin nidus and then proliferate [15]. Microbial growth results in activation of the extrinsic coagulation pathway, monocytes release a myriad of pro-inflammatory cytokines and there is increased expression of fibronectin on the surface of the endothelial cells with resultant formation of a vegetation.
The vegetation grows further, with subsequent embolization and continued bacteraemia, if the host is unable to contain the infection [16].
Right-sided infective endocarditis usually presents with fever, persistent bacteraemia and septic emboli to the lungs. Initial presentation may comprise haemoptysis, cough or chest pain. Peripheral embolization must alert one to the presence of concomitant left-sided endocarditis or a shunt. Right heart failure is a result of both pressure and volume overload from pulmonary hypertension or organic tricuspid regurgitation or rarely obstruction of the tricuspid orifice by a vegetation [17, 18].
Pulmonary septic emboli may be complicated by pulmonary infarction, abscess, pneumothorax, and purulent pulmonary effusion [17] (Figures 1 and 2).
An anterior-posterior chest X-ray showing increased cardiothoracic index with areas of alveolar opacification involving both lung fields likely representing septic embolization and abscess formation in the lungs.
Multiple areas of consolidation suggestive of infarction and dilated right heart chambers on a CT scan of the chest of a patient with history of right-sided infective endocarditis due to intravenous drug abuse.
It is important to note that patients with RSIE do not always have an audible murmur of tricuspid regurgitation [13]. Other features unique to this group of patients with IE are the presence of co-infections with HIV, hepatitis C and hepatitis B infections, which complicate their clinical management and adversely affect their outcomes. A high degree of suspicion of IE must be maintained in IVDA as their clinical assessment can be quite challenging, especially in those who do not manifest the classic clinical features.
Additionally, the sensitivity and specificity of the modified Duke’s criteria in right-sided endocarditis has not been studied. Inclusion of septic pulmonary infarcts as a minor criteria in the modified Duke’s criteria may therefore be inappropriate [19].
In addition to the above mentioned clinical features and positive blood cultures, transthoracic echocardiography (TTE) greatly aids in establishing a diagnosis of IE, especially in cases with equivocal clinical presentation. TTE allows easy visualisation of vegetations on the anteriorly located tricuspid valve and associated tricuspid regurgitation (Figures 3 and 4) [20, 21].
Modified apical four-chamber view showing multiple vegetations on the tricuspid valve (arrow) with a dilated right atrium and right ventricle.
Colour Doppler ultrasound showing severe tricuspid regurgitation.
A transesophageal echocardiography may be required in detection of vegetations on the pulmonary valve and for exclusion of left-sided valve involvement [22]. The Eustachian valve must be screened for presence of vegetations.
The initial antimicrobial therapy should take into account four factors: (1) suspected organism (2) type of drug (3) the solvent used by the addict and (4) the location of Infection [17].
Empirical therapy in acute severely ill patients must consist of ampicillin and cloxacillin with gentamycin or vancomycin with gentamycin (in patients allergic to penicillin) [17]. Staphylococcus aureus must always be covered. Anti-pseudomonas agent must be added in a pentazocine drug addict. If an IVDA gives a history of brown heroin use mixed with lemon juice then an anti-fungal agent must be added due to a high risk of candida septicaemia. Anti-microbial therapy can be de-escalated once the specific causative organism is isolated on blood cultures.
Due to reluctance of IVDA for prolonged hospital admission and the concerns related to their discharge on intravenous antibiotic therapy, a few studies have studied the possibility of treating IE in these patients with short course antibiotic therapy [23].
A 2 week treatment regimen has been advocated in non-complicated isolated tricuspid valve endocarditis. These patients must have low risk features such as good response to therapy, methicillin sensitive Staphylococcus aureus, small vegetation size (less than 20 mm), no features of peripheral embolization, absence of metastatic infection, lack of involvement of left-sided valves or prosthetic valve and absence of a severely immunosuppressed state. In such cases, a short 2 week course of intravenous cloxacillin or oxacillin alone may be used [24]. These patients must be closely followed up and the response to therapy must be assessed.
In complicated cases a 4–6 week course of intravenous therapy must be utilised. These include situations where there is poor response to antibiotic therapy, large vegetation size (>20 mm), septic emboli, use of penicillinase non-resistant antibiotics, and a severely immunosuppressed state such as HIV with a CD4 count less than 200cell/ml and associated involvement of left-sided valves [25, 26, 27].
Due to a high rate of recurrent IE in IVDA, surgery should only be considered in the following situations: (1) intractable right-sided heart failure with poor response to diuretics; (2) persistent bacteraemia despite use of appropriate antimicrobial therapy; and (3) large vegetation size of greater than 20 mm that do not diminish in size after repeated episodes of pulmonary emboli [25, 28, 29].
In general the outcomes of patients with IVDA related IE have been poor post surgery. A substantially high long term mortality has been reported for IE related surgery in IVDA compared to non-drug users [30, 31, 32].
In HIV-infected IVDAs with IE cardiac surgery does not worsen the outcome of either the IE or the HIV [17]. Patients with advanced HIV infection with severe immunosuppression. However, valve replacement surgery may have unacceptably high risks in selected patients with advanced HIV infection, low CD4 counts, and either a history of failed antiretroviral therapy or ongoing drug abuse that precludes therapy with antiretroviral agents [33].
The most commonly performed surgery for tricuspid valve endocarditis includes valvectomy, valve replacement or repair [34]. Valve repair is advocated by some studies but repair has not proven to be superior to either valve replacement or valvectomy. In a few cases of RSIE valvectomy may be performed initially followed by subsequent bioprosthetic valve replacement once the infection has subsided and drug use discontinued. Pulmonary valve rarely requires replacement except in extreme cases of valve destruction. In cases where pulmonary valve replacement is deemed suitable, a homograft is preferred.
Overall, IVDA with RSIE have a lower mortality than those with left-sided infective endocarditis [14, 24, 35, 36, 37, 38, 39]. In one study the mortality was noted to be 6% [40]. Factors associated with high mortality included a large vegetation size (>20 mm) and a fungal aetiology [41, 42].
In general patients with HIV do not have a poor outcome, except those with CD4 count <200 cells/ml. The major reason for repeat hospitalisation and recurrent endocarditis in IVDA is related to persistent use of drugs [30, 43, 44].
Finally, management of RSIE related to IVDA poses some ethical dilemmas. From the limited available literature, surgery should be offered for patients with surgical indications, with a first episode of IE in IVDAs, who are willing to undergo rehabilitation. If the patient presents with a second episode of IE due to recurrent IVDA, the decision to re-operate the patient, if indicated, is complex. It should be individualised and discussed by the endocarditis team. It is reasonable to decline further surgical intervention in this group, especially in resource-limited settings [45].
The Duffy blood group system, ISBT number 008/symbol (FY), was published for the first time in 1950 when anti-Fya was identified in a suspected hemolytic transfusion reaction in a 43-year-old patient with hemophilia who received 3 packed red blood cell (PRBC) units for treatment of spontaneous bleeding and who developed jaundice 1 day after transfusion [1, 2]. Approximately, 1 year later, anti-Fyb was discovered in a postpartum blood sample from a patient who gave birth to her third child [3].
\nChromosome 1 has both FY and RH gene loci. The FY locus is located on the long arm at position 1q22-q23 where it consists of two exons distributed over 1.5 kbp of gDNA, whereas RH resides on the short arm. The Duffy system is N-glycosylated multi-pass transmembrane glycoprotein (Figure 1) [4] also known as the atypical chemokine receptor 1 (ACKR1, CD234). The protein is composed of 336 amino acids. There are two possible Duffy mRNAs which are translated from the Duffy antigen gene, a less abundant α form (338 amino acids) and a major β form (336 amino acids) which differ by 2 amino acids in the N-terminus. Approximately 6000–13,000 copies of the Duffy protein are found on the surface of RBCs [5].
\nThe predicted seven-transmembrane domain structure of the Duffy protein. The amino acid change responsible for Fya/Fyb polymorphism, the mutation responsible for Fyx, and the glycosylation sites and the regions where Fy3 (and Fy6) map are indicated (reproduced with permission).
The Duffy blood group includes six known antigens that differ by amino acid sequence. The Duffy antigen prevalence varies between racial groups.
\nACKR1 (previously known as DARC) is a receptor for a variety of chemokines, including interleukin-8, monocyte chemotactic protein-1, and melanoma growth stimulatory activity. Also, this glycoprotein is a receptor for Plasmodium vivax and Plasmodium knowlesi; thus red cells with Fy(a-b-) phenotype are resistant to invasion by these malarial species. Antibodies formed against the Duffy antigens show a dosage effect and are a cause of both hemolytic transfusion reactions and hemolytic disease of fetus and newborn. The Duffy protein is also found on the endothelial cells of capillary and postcapillary venules, the epithelial cells of kidney collecting ducts, lung alveoli, and Purkinje cells of cerebellum [6].
\nThere are six known antigens with four main phenotypes; Fy(a+b+), Fy(a−b+), Fy(a+b−), and Fy(a−b−) (Table 1) [5]. The most common antigens are, two polymorphic and antithetical, Fya (FY1) and Fyb (FY2) which differ by one amino acid at position 42 on the extracellular domain, with glycine resulting in Fya expression and aspartic acid resulting in Fyb expression [5, 7]. They are sensitive to destruction when RBCs are treated with proteolytic enzymes such as papain or ficin, whereas, there is no RBCs destruction with trypsin treatment [8].
\nRed cell phenotype | \nPrevalence (%) | \nAllele | \n|
---|---|---|---|
Caucasians | \nBlacks | \n||
Fy (a+b−) | \n17 | \n9 | \nFY*01/FY*01 or FY*A/FY*A | \n
Fy (a−b+) | \n34 | \n22 | \nFY*02/FY*02 or FY*B/FY*B | \n
Fy (a+b+) | \n49 | \n1 | \nFY*A/FY*B | \n
Fy (a−b−) | \nRare | \n68 | \nFY*/N.01–05, FY*/N.01–02\n‡\n\n | \n
Fy3\n | \n100 | \n32 | \n\n |
Fy5\n | \n99.9 | \n32 | \n\n |
Fy6\n | \n100 | \n32 | \n\n |
Duffy blood group system phenotypes and prevalence. Reproduced with permission and modification.
Nomenclature pending approval by the ISBT working party on terminology for red cell surface antigens.
Fya antigen has a prevalence of 66% in Caucasians, 10% in Blacks, and 99% in Asians. It has been identified on fetal RBCs as early as 6 weeks gestation and reaches adult levels in approximately 12 weeks after birth. Fyb has a prevalence of 83% in Caucasians, 23% in Blacks, and 18.5% in Asians. It is expressed on cord blood cells. Fy3 antigen is expressed in 100% of Caucasians, 32% of Blacks, and 99.9% of Asians. It is also expressed on cord cells and demonstrates increased expression after birth. Fy5 antigen is expressed on 32% of Blacks and 99.9% of Caucasians and Asians. It is not expressed on Rh null RBCs. Fy6 is expressed in 100% of most populations and 32% of Blacks. The Fy(a–b–) phenotype is the major phenotype in approximately 70% Blacks, but is very rarely found in other populations. This phenotype is characterized by the absence of the Fyb antigen on RBCs and its presence on non-erythroid cells. Duffy mRNA is not detected in the bone marrow of Fy(a–b–) individuals; however, it is detected in other tissues including the colon, lung, and spleen. This unique phenotype is caused by a single amino acid substitution at position 46 in the Duffy (Fyb) gene. This mutation impairs the promotor activity in erythroid cells by disrupting the binding site for GATA1 erythroid transcription factor. Furthermore, some individuals with this phenotype do not make anti-Fyb. This is believed to be due to a mutation in the, erythroid promoter, GATA-1 binding motif. Interestingly, the same Fy(a–b–) phenotype rarely found in Caucasians is characterized by absence of Duffy antigens expression in both erythroid and non-erythroid tissues due to possibly presence of mutations which prevent formation of Duffy protein. These individuals can form anti-Fy3. The have high prevalence antigens; Fy3, Fy5, and Fy6 are conformational epitopes as opposed to specific sequence epitopes with Fy5 hypothesized to be a combined conformational epitope of Duffy and Rh protein [9, 10, 11, 12].
\nAnti-Fya and -Fyb are clinically significant RBC alloantibodies which can cause immediate and delayed hemolytic transfusion reactions (HTRs) as well as hemolytic disease of the fetus and newborn (HDFN). They often result from previous exposure such as after transfusion or pregnancy. They are not usually naturally occurring. The Duffy antibodies are predominantly of the IgG subclass whereas the IgM form is rare.
\nThe mechanism of extravascular hemolysis (EH) in both HDFN and HTR is similar. In HDFN, the mother lacks a certain red cell antigen which the fetus is positive for, thus the mother is allo-immunized (i.e., made a new antibody) during the first pregnancy. If she gets exposed to the same antigen in subsequent pregnancy (ies), the fetus (es) is/are at risk of HDFN. Similarly, if a patient lacks a certain red cell antigen but receives red cell transfusion with a unit that has such antigen, the patient is at risk for allo-immunziation after the transfusion and HTR in subsequent transfusion (s). EH is typically induced by IgG red cell antibodies. EH consists of consumption of antibody and/or C3b-bound red cells by phagocytes in the reticuloendothelial system (RES) causing a delayed hemolytic transfusion reaction (DHTR). DHTRs can be clinically significant leading to morbidity and possibly mortality. To avoid DHTR, patients with known clinically significant antibodies, receive red cell units that lack antigen (s) to their the cognate antibody (ies). The Duffy antibodies are usually associated with a moderate DHTR and mild HDFN [13].
\nAnti-Fya is identified more than anti-Fy3, anti-Fy5, or anti-Fyb. Fya is 20 times more immunogenic than Fyb. Some of anti-Fya can bind and activate complements [14]. Anti-Fy3 is also clinically significant antibody which can cause mild HDFN and HTRs. Serologically, it can react with enzyme treated Fy(a+) or Fy(b+) RBCs, but fails to react with Fy(a−b−) RBCs [15]. Anti-Fy4 shows lack of consistent test results. It was found to be reactive with Fy(a−b−), some Fy(a+b−), some Fy(a−b+) RBCs but shows no reaction with Fy(a+b+) RBCs [16]. Anti-Fy5 reacts with enzyme treated Fy(a+) or Fy(b+) RBCs with no reaction with Fy(a−b−) RBCs or Rh null RBCS. It has been reported in sickle cell patients with delayed HTRs in the presence of other clinically significant alloantibodies [17]. A human anti-Fy6 has not been identified [18].
\nThe Duffy glycoprotein can bind to a variety of chemokines and is known commonly as the Duffy antigen receptor for chemokines (DARC) or more recently atypical chemokine receptor 1 (ACKR1). Chemokines are proteins secreted by immune cells as a mean to communicate signals to guide their interactions. The exact function of DARC is not fully clear. One postulated function is that DARC permits erythrocyte to act a chemokine scavenger to limit leukocyte activation. The importance of this function in inflammatory diseases is not well established [6, 19].
\nThe Duffy glycoprotein plays an important role in malaria transmission by acting as the erythroid receptor for Plasmodium vivax through binding to the Fy6 epitope (previously known as P. vivax Duffy-binding protein (PvDbp)) and for Plasmodium knowlesi. Individuals with Fy(a−b−) phenotype were resistant to parasitic invasion in a study performed on 11 volunteers, whereas those who contracted malaria were Fy(a+) or Fy(b+). Fy6 is present on all erythroid cells with an Fy(a+) or Fy(b+) phenotype. Thus it is absent on red cells with Fy(a−b−) phenotype. In west Africa, individuals with Fy(a−b−) phenotype are found in greater frequency than in areas where P. vivax is absent. The protective effect of Fy(a−b−) phenotype does not extend to P. falciparum which can infect red cells of all Duffy phenotype [20].
\nI want to thank the department of Pathology at the University of Chicago, Chicago, IL, United States.
\nThe author declares no conflict of interest.
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