Histomorphological criteria for spongiosis, epidermal lymphocytes, basal cell vacuolation and melanophages.
\r\n\tThere are different types of multiple pregnancies: fraternal twins, identical twins, triplets, and higher-order multiples. Symptoms of multiple pregnancies are larger uterus than expected for the date in pregnancy, increased morning sickness, increased appetite, and excessive weight gain. In this book, we will examine the clinical aspects of multiple pregnancies and management. Also, we will examine the management of cases of twins including antenatal care, delivery, and postpartum.
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Goudos",coverURL:"https://cdn.intechopen.com/books/images_new/5436.jpg",editedByType:"Edited by",editors:[{id:"171056",title:"Dr.",name:"Sotirios",surname:"Goudos",slug:"sotirios-goudos",fullName:"Sotirios Goudos"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}],publishedBooksByAuthor:[]},onlineFirst:{chapter:{type:"chapter",id:"69700",title:"Histopathology and Molecular Pathology of Vitiligo",doi:"10.5772/intechopen.84258",slug:"histopathology-and-molecular-pathology-of-vitiligo",body:'Vitiligo is a common acquired, idiopathic, progressive disorder which is characterized by the development of depigmented milky white macules of variable sizes. These often enlarge and coalesce to form extensive areas of leukoderma [1, 2, 3]. It equally affects both sexes with a worldwide prevalence of 0.1–2% [4]. It is a psychologically devastating and frequently resistant to treatment [5, 6]. The basic defect in vitiligo is a selective destruction of functional melanocytes [7].
The role of histopathology in the diagnosis of vitiligo is not yet fully established. So much so that routinely in these cases biopsy is not performed. The diagnosis is made primarily on clinical grounds.
Vitiligo is a multifactorial disorder [8, 9]. In its genesis both genetic and non-genetic factors are believed to play a role. It is observed that clinically no two patients of vitiligo are alike. This suggests that etiology also varies among different patients. Due to the observed variation in clinical manifestations of the disease, it seems likely that etiology of vitiligo may differ among patients [10]. These several theories have been combined into the convergence theory [11] which is currently the most accepted theory.
Briefly in the earliest theory, it was proposed by Lerner that vitiligo was neural in origin [12]. This theory could explain the segmental form of vitiligo which follows dermatomal distribution and is associated with hyperhidrosis and emotional disturbances. In another study the role of sympathetic nervous system in vitiligo was studied [13]. It was observed that the cutaneous blood flow in the lesional skin was three times higher than the normal skin in cases of segmental vitiligo. However, in other cases of non-segmental vitiligo, this was not observed.
Studies on the expression of neural proteins like neuropeptide Y (NPY), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP) and polyclonal general neuronal marker (PGP) have shown variable results. In one study NPY expression was found to be increased in cases of segmental vitiligo [14]. It is proposed that precipitating factors like stress lead to increased NPY expression [15].
But this theory failed to explain the other forms of vitiligo. For that matter generalized or non-segmental vitiligo is better explained by autoimmune hypothesis. In previously done studies, antibodies against various targets like tyrosine hydroxylase, melanin-concentrating hormone receptor-1 (MCHR1), tyrosinase [16] and pigment cell surface antigens [17] have been demonstrated. In a study carried out to evaluate the various immunoglobulins, it was observed that 80% of active vitiligo patients showed the presence of IgG and IgM against melanocytes [17]. Other studies have shown the presence of anti-thyroglobulin antibodies, antithyroid antibodies, anti-thyroperoxidase and anti-smooth muscle antibody in these cases [18, 19].
Besides humoral immunity, cell-mediated immunity may also play an important role. Immunohistochemical examination of perilesional skin in vitiligo patients showed increased CD8:CD4 ratio and HLA-DR production along suprabasal and basal keratinocytes. Macrophages were found to be quite numerous [20]. However, not only the immune cells and antibodies but expression of various cytokines is also increased. Chief among these which have been studied are tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and IL-10 [21, 22]. IL-17 has also been shown to be significantly increased in cases of vitiligo [23].
The redox (reduction–oxidation) state of vitiliginous patients has been studied by many authors. These studies have shown increased serum levels of selenium, superoxide dismutase (SOD) and malondialdehyde (MDA) [24, 25]. Increased levels of these substances indicate the presence of oxidative stress in vitiligo. Increased levels of tetrahydrobiopterin [26] and xanthine oxidase [27] leading to increased levels of H2O2 may also be contributory.
Few authors have pointed towards the role of zinc-α2-glycoprotein (ZAG) in the pathogenesis of vitiligo. They hypothesize that lack of ZAG causes impaired melanocytic adhesion to other cells in the epidermis [28, 29]. The efficacy of zinc in the treatment of vitiligo may be due to its ability to precipitate ZAG at the site of vitiligo [30].
The role of viral infection in vitiligo has been proposed by certain authors. The potential candidates include hepatitis C virus (HCV) [31], cytomegalovirus (CMV) [32], Epstein–Barr virus (EBV), hepatitis e virus, herpes virus and HIV [33]. However, the evidence available is scant and not conclusive enough to attribute a significant role for viral agents in vitiligo.
The intrinsic theory states that in vitiligo there is loss of melanocytes due to various abnormalities which lead to increased apoptosis [34] and accelerated cell senescence [35, 36]. Studies done previously have shown various abnormalities in the melanocytes including cytoplasmic vacuolization, DNA marginalization, dendrite loss and detachment [36, 37]. The evidence in favor of increased apoptosis in vitiligo includes reduced expression levels of the antiapoptotic proteins Bcl-2 and FLIP in vitiliginous skin as compared to normal skin [34]. On the other hand, marked increase in the expression of proapoptotic factors such as Bax and p53 along with the various caspases has also been observed [34].
The melanocytorrhagy theory states that in vitiligo there is chronic melanocyte detachment and loss caused by trauma and other stressors which include catecholamines, free radicals or autoimmune elements [38].
However, the consensus opinion of majority of experts is that vitiligo occurs due to convergence of these various pathways [39]. These are also depicted in (Figure 1). The author also is in agreement with this view; however, it is likely that in various subtypes of vitiligo the relative contribution of these pathways may vary. For example, in segmental vitiligo the neural theory may be more relevant than the other theories, whereas the same may not hold true for vitiligo vulgaris.
Pathogenesis of vitiligo.
In order to understand the histopathology of vitiligo, it is essential to first understand the concept of
The basic histopathological finding in vitiligo is the absence of functional melanocytes in the basal layer of the epidermis (Figure 2) [42, 43, 44]. This absence can also be demonstrated by using special stains like Fontana-Masson (Figure 3) [45]. Immunohistochemistry for melanocyte-specific markers like HMB-45 and Melan-A and electron microscopy can also be performed for this purpose.
Vitiligo showing the absence of melanocytes in basal layer of the epidermis (H&E, X400).
(a) Fontana-Masson showing the presence of melanin pigment and melanocytes in the epidermis (X400). (b) Fontana-Masson showing the absence of melanin pigment and melanocytes in the epidermis (x400).
Other changes that have been observed include degenerative changes in the nerves and adnexa like sebaceous glands and hair follicles especially in long-standing cases [46].
In the margins of lesions especially early lesions, often inflammatory cells are seen. Principally, these cells comprise of CD4+ and CD8+ T cells [47]. These cells have been shown to demonstrate melanocyte-specific cytotoxicity [48]. At the margins of the lesions, melanocytes have been observed to show morphological changes like cellular enlargement, cytoplasmic vacuolization and long dendritic processes [29].
However, usually skin biopsy is not performed for making the diagnosis as it is primarily a clinical diagnosis. The cornerstone of its management is correct categorization of a case into its two broad types—stable and unstable vitiligo. This distinction is at present based mainly on clinical criteria because the histopathological features are not fully established. In a study carried out by the author, a reliable and systematic approach towards this diagnostic challenge has come up [49]. In that study the biopsies (3-mm punch) were taken from the margin of the active lesion.
The author recommends that while evaluating biopsies from cases of vitiligo histopathological examination should be primarily focused on evaluating five histopathological variables—spongiosis, epidermal lymphocytes, basal cell vacuolation, dermal lymphocytes and melanophages (Figure 4). The morphological criteria used to assess these parameters are listed in Table 1. All the cases are then scored using a scoring system devised by the authors Table 2, and the recommended diagnoses based on these scores are shown in Table 3. The counting for dermal lymphocytes was done in high power (x400) of a Nikon microscope. The scoring system can be applied to both segmental and non-segmental vitiligo. Adoption of a systematic reporting system brings more consistency and objectivity in the diagnosis.
(a) Spongiosis (H&E, x400). (b) Intraepidermal lymphocytes (H&E, x400). (c) Basal cell vacuolation (H&E, x400). (d) Dermal lymphocytosis (H&E, x200). (e) Dermal melanophages (H&E, x400).
S. no. | Histological feature | Criteria |
---|---|---|
1 | Spongiosis | Presence of at least one focus showing intercellular oedema in the epidermis |
2 | Epidermal lymphocytes | Presence of at least one lymphocyte in the epidermis |
3 | Basal cell vacuolation | Presence of at least one focus showing basal cell degeneration in the form of vacuolation |
4 | Melanophages | Presence of at least one focus in the superficial reticular dermis showing melanophages |
Histomorphological criteria for spongiosis, epidermal lymphocytes, basal cell vacuolation and melanophages.
S. no. | Histological feature | Observation | Score |
---|---|---|---|
1 | Spongiosis | Present/absent | 1/0 |
2 | Epidermal lymphocytes | Present/absent | 1/0 |
3 | Basal vacuolation | Present/absent | 1/0 |
4 | Dermal lymphocytes >100 | Present/absent | 1/0 |
5 | Melanophages | Present/absence | 1/0 |
Vitiligo histological scoring system.
S. no. | Total score | Diagnosis |
---|---|---|
1. | 5 | Unstable vitiligo |
2. | 4 | Unstable vitiligo |
3. | 3 | Favours unstable, clinical correlation required |
4. | 2 | Favours stable, clinical correlation required |
5. | 0–1 | Strongly favours stable vitiligo, clinical correlation essential |
Final recommended diagnostic categories based on score.
Vitiligo is a common skin disorder which is characterized by the presence of depigmented milky white macules of variable sizes. Although there are various theories on its etiopathogenesis, the consensus opinion is that vitiligo occurs due to convergence various pathways. The basic histopathological finding in vitiligo is the absence of functional melanocytes in the basal layer of the epidermis. However, in order to evaluate for stability, the histopathological examination should be primarily focused on evaluating spongiosis, epidermal lymphocytes, basal cell vacuolation, dermal lymphocytes and melanophages. It is recommended to score these parameters, and the final report should incorporate recommended diagnosis based on the score. This will bring more objectivity and consistency in reporting these biopsies.
The author wishes to acknowledge the help provided by Dr. Niti Khunger, Consultant, Dermatology, VMMC & Safdarjung Hospital, and Dr. Pallavi Mishra, Resident, VMMC & Safdarjung Hospital.
The author wishes to declare that there are no conflicts of interest.
None declared.
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\n\nMetadata for all publications is also automatically deposited in IntechOpen's OAI repository, making them available through the Open Access Infrastructure for Research in Europe's (OpenAIRE) search interface further establishing our compliance.
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virus (AAV) has been isolated from numerous vertebrate species since 1966. Besides its wide and promiscuous tropism, AAV infection does not result in considerable toxicity or pathogenicity and is capable of achieving adequate and long-term levels of gene transfer, especially following generation of the AAV recombinant variant: rAAV. Due to these properties, rAAV has gained special attention as a viral vector for gene therapy in the last decade. Currently, there are 130 clinical trials taking place worldwide for several diseases testing the safety and efficacy profiles of rAAV. During preclinical and clinical studies, several challenges have arisen in terms of reaching the full therapeutic potential of rAAV, such as efficient delivery of the virus in a targeted and specific manner to a desired tissue. Importantly, the development of immune responses towards the viral capsids poses an obstacle to rAAV applicability in the clinical setting. Numerous approaches have been developed in order to tailor an optimized therapeutic virus for treating specific diseases, including the use of different AAV serotypes or the creation of recombinant capsid variants with distinctive transduction and immunological profiles. This chapter reviews current information on rAAV clinical trials and the potential for combining rAAV platform with other technologies, such as induced pluripotent cells and gene editing.",book:{id:"4754",slug:"gene-therapy-principles-and-challenges",title:"Gene Therapy",fullTitle:"Gene Therapy - Principles and Challenges"},signatures:"Melisa A. Vance, Angela Mitchell and Richard J. Samulski",authors:[{id:"174649",title:"Ph.D.",name:"Melisa",middleName:null,surname:"Vance",slug:"melisa-vance",fullName:"Melisa Vance"},{id:"175144",title:"Dr.",name:"R. Jude",middleName:null,surname:"Samulski",slug:"r.-jude-samulski",fullName:"R. 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Numerous approaches have been developed in order to tailor an optimized therapeutic virus for treating specific diseases, including the use of different AAV serotypes or the creation of recombinant capsid variants with distinctive transduction and immunological profiles. This chapter reviews current information on rAAV clinical trials and the potential for combining rAAV platform with other technologies, such as induced pluripotent cells and gene editing.",book:{id:"4754",slug:"gene-therapy-principles-and-challenges",title:"Gene Therapy",fullTitle:"Gene Therapy - Principles and Challenges"},signatures:"Melisa A. Vance, Angela Mitchell and Richard J. Samulski",authors:[{id:"174649",title:"Ph.D.",name:"Melisa",middleName:null,surname:"Vance",slug:"melisa-vance",fullName:"Melisa Vance"},{id:"175144",title:"Dr.",name:"R. Jude",middleName:null,surname:"Samulski",slug:"r.-jude-samulski",fullName:"R. Jude Samulski"},{id:"178296",title:"Dr.",name:"Angela",middleName:null,surname:"Mitchell",slug:"angela-mitchell",fullName:"Angela Mitchell"}]},{id:"63034",title:"Mitochondrial Dysfunction Associated with Doxorubicin",slug:"mitochondrial-dysfunction-associated-with-doxorubicin",totalDownloads:1634,totalCrossrefCites:6,totalDimensionsCites:13,abstract:"Cancer prevalence is scaling up each year. Anthracycline groups are still the best chemotherapeutic agent. The most popular anticancer drug in the group is doxorubicin (DOX). Unfortunately, DOX has potent toxicity on noncancerous tissues, e.g., heart, kidneys, etc. However, it is well documented that the severest toxicity of the drug affects heart tissue. Of course, some reasons have been suggested why and/or how the heart is so vulnerable to toxicity. The primary mechanism responsible for DOX’s cardiospecific toxicity remains unidentified so far; however, mitochondrial dysfunction induced by DOX is now considered one of the leading reasons for DOX’s toxicities and undesired side effects. Mitochondrial reactive oxygen production in the heart is a significant contributor to developing mitochondrial dysfunction-exposed DOX based on a variety of evidence. The objective of this review chapter is to critically evaluate and highlight the role of mitochondria in the development of DOX-induced cardiotoxicity.",book:{id:"6060",slug:"mitochondrial-diseases",title:"Mitochondrial Diseases",fullTitle:"Mitochondrial Diseases"},signatures:"Celal Guven, Yusuf Sevgiler and Eylem Taskin",authors:[{id:"192567",title:"Prof.",name:"Eylem",middleName:null,surname:"Taskin",slug:"eylem-taskin",fullName:"Eylem Taskin"},{id:"195229",title:"Dr.",name:"Celal",middleName:null,surname:"Guven",slug:"celal-guven",fullName:"Celal Guven"},{id:"206996",title:"Prof.",name:"Yusuf",middleName:null,surname:"Sevgiler",slug:"yusuf-sevgiler",fullName:"Yusuf Sevgiler"}]}],onlineFirstChaptersFilter:{topicId:"186",limit:6,offset:0},onlineFirstChaptersCollection:[{id:"81348",title:"Mouse Models to Understand Mutagenic Outcomes and Illegitimate Repair of DNA Damage",slug:"mouse-models-to-understand-mutagenic-outcomes-and-illegitimate-repair-of-dna-damage",totalDownloads:14,totalDimensionsCites:0,doi:"10.5772/intechopen.103929",abstract:"Maintenance of genome integrity is critical to prevent cell death or disease. Illegitimate repair of chromosomal DNA breaks can lead to mutations and genome rearrangements which are a well-known hallmark of multiple cancers and disorders. Endogenous causes of DNA double-strand breaks (DSBs) include reactive oxygen species (ROS) and replication errors while exogenous causes of DNA breaks include ionizing radiation, UV radiation, alkylating agents, and inhibitors of topoisomerase II (Top2). Recent evidence suggests that a growing list of environmental agents or toxins and natural dietary compounds also cause DNA breaks. Understanding the consequences of exposure to a broad spectrum of DSB-inducing agents has significant implications for understanding mutagenicity, genome stability and human health. This chapter will review in vivo mouse models designed to measure DNA damage and mutagenicity, and illegitimate repair of DNA DSBs caused by exposure to environmental agents.",book:{id:"11348",title:"Mutagenesis and Mitochondrial-Associated Pathologies",coverURL:"https://cdn.intechopen.com/books/images_new/11348.jpg"},signatures:"Kiran Lalwani, Caroline French and Christine Richardson"},{id:"79374",title:"Mitochondrial Cytopathies of the Renal System",slug:"mitochondrial-cytopathies-of-the-renal-system",totalDownloads:71,totalDimensionsCites:0,doi:"10.5772/intechopen.96850",abstract:"Mitochondria are major intracellular organelles with a variety of critical roles like adenosine triphosphate production, metabolic modulation, generation of reactive oxygen species, maintenance of intracellular calcium homeostasis, and the regulation of apoptosis. Mitochondria often undergo transformation in both physiological and pathological conditions. New concepts point that mitochondrial shape and structure are intimately linked with their function in the kidneys and diseases related to mitochondrial dysfunction have been identified. Diseases associated with mitochondrial dysfunction are termed as “mitochondrial cytopathies”. Evidence support that there is a role of mitochondrial dysfunction in the pathogenesis of two common pathways of end-stage kidney disease, namely, chronic kidney disease (CKD) and acute kidney injury (AKI). Mitochondrial cytopathies in kidneys mainly manifest as focal segmental glomerular sclerosis, tubular defects, and as cystic kidney diseases. The defects implicated are mutations in mtDNA and nDNA. The proximal tubular cells are relatively vulnerable to oxidative stress and are therefore apt to suffer from respiratory chain defects and manifest as either loss of electrolyte or low-molecular-weight proteins. Patients with mitochondrial tubulopathy are usually accompanied by myoclonic epilepsy and ragged red muscle fibers (MERRF), and Pearson’s, Kearns-Sayre, and Leigh syndromes. The majority of genetic mutations detected in these diseases are fragment deletions of mtDNA. Studies have shown significantly increased ROS production, upregulation of COX I and IV expressions, and inactivation of complex IV in peripheral blood mononuclear cells of patients with stage IV–V CKD, thereby demonstrating the close association between mitochondrial dysfunction and progression to CKD. Furthermore, the mechanisms that translate cellular cues and demands into mitochondrial remodeling and cellular damage, including the role of microRNAs and lncRNAs, are examined with the final goal of identifying mitochondrial targets to improve treatment of patients with chronic kidney diseases.",book:{id:"11348",title:"Mutagenesis and Mitochondrial-Associated Pathologies",coverURL:"https://cdn.intechopen.com/books/images_new/11348.jpg"},signatures:"Lovelesh K. Nigam, Aruna V. Vanikar, Rashmi Dalsukhbhai Patel, Kamal V. Kanodia, Kamlesh Suthar and Umang Thakkar"},{id:"75895",title:"Maneuvering Mitochondria for Better Understanding of Therapeutic Potential of mtDNA Mutation",slug:"maneuvering-mitochondria-for-better-understanding-of-therapeutic-potential-of-mtdna-mutation",totalDownloads:114,totalDimensionsCites:0,doi:"10.5772/intechopen.96915",abstract:"Heterogeneity of mitochondrial diseases in terms of genetic etiology and clinical management makes their diagnosis challenging. Mitochondrial genome, basic mitochondrial genetics, common mutations, and their correlation with human diseases is well-established now and advances in sequencing is accelerating the molecular diagnostics of mitochondrial diseases. Major research focus now is on development of mtDNA intervention techniques like mtDNA gene editing, transfer of exogenous genes (sometimes even entire mtDNA) that would compensate for mtDNA mutations responsible for mitochondrial dysfunction. Although these genetic manipulation techniques have good potential for treatment of mtDNA diseases, research on such mitochondrial manipulation fosters ethical issues. The present chapter starts with an introduction to the factors that influence the clinical features of mitochondrial diseases. 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OXPHOS occurs in the inner membrane of the mitochondrion and involves 5 protein complexes that sequentially undergo reduction-oxygen reactions ultimately producing adenosine triphosphate (ATP). Tissues with high metabolic demand such as lungs, central nervous system, peripheral nerves, heart, adrenal glands, renal tubules and the retina are affected preferentially by this critical role in energy production by mitochondrial disorders. Eye-affected mitochondrial disorders are always primary, but the role of mitochondrial dysfunction is now best understood in acquired chronic progressive ocular diseases. Recent advances in mitochondrial research have improved our understanding of ocular disorders. 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