**2. Penile cancer**

Penile malignancies are thought to arise from the accumulation of multiple mutations that may occur as consequence of progressive genetic instability. This intricate process of genetic instability may be caused by environmental factors, such as history of intense smoking, penile tears, phimosis, and poor genital hygienic habits. (Chaux & Cubilla 2012). In addition, a recent study conducted by Chaux et al. (2011) also described the poor education, penile chronic inflammation, genital warts and Human papillomavirus (related to number of sexual partners during lifetime) as environmental factors to malignant transformation.

There is a worldwide geographic difference in occurrence of penile malignancies that could be caused by differences in socio-economic status, cultural and religious conditions (Bleeker et al. 2009, Chaux & Cubilla, 2012). The higher incidences are frequent in tropical or subtropical regions of Latin America, Asia and Africa but have uncommon incidence rates in Europe, Japan, USA and Israel (Cubilla 2009). Recently, were reported higher incidence rates in underdeveloped regions such as Africa, South America, and Asia (2-4/100000 inhabitants) as compared with North America (United States) and Europe (0.3-1/100000 inhabitants) (Chaux & Cubilla, 2012). Pow-Sang et al. (2010) also described the penile malignancies prevalence rate among different populations. Prevalence rates in developed countries as Israel (0.1/100 000) and United States (0.3-1.8/100 000) and interestingly, compared with underdeveloped coun‐ tries such as Uganda (2.8/100 000) and Brazil (1.5-3.7/100 000). Once again confirming the disease geographical difference and the influence of country development.

Squamous cell Carcinoma represents vast majority of histological subtype of primary penile malignancies with heterogeneous features due to differences in morphology pathogenesis and prognosis (Hakenberg & Protzel, 2012; Stankiewicz et al., 2012; Syed et al., 2012). Knowledge of origin and progression of penile squamous cell carcinoma depends on an intricate relation between anatomy and histopathology.

cell cycle checkpoints and to entrance in S1 phase of cell cycle, leading to disruption of normal

222 Human Papillomavirus and Related Diseases – From Bench to Bedside A Diagnostic and Preventive Perspective

Following cell division, infected cells leave the basal layer, migrate towards the suprabasal regions and begin to differentiate. Increased understanding of cervical pathogenesis has led to confirmation of HPV as an etiological agent for several cancers and consequently to the development of preventive vaccines targeting HPV antigens for the control of HPV-related cancers. HPVvaccinewasdevelopedas a result ofthe achievement of core technologies,that are able to produce virus-like particles (VLPs), which, in turn, are able to mimic the natural virus andelicithigh-titersofvirusneutralizingantibodies.Withtheprogressthroughadvancedstages of clinical trials and further exploration of combinatorial strategies, there is a great promise for significantadvancesalsointhefieldoftherapeuticHPVvaccinedevelopment,notonlytocervical cancer, but other several malignancies related to HPV infection. Moreover, in this chapter we discuss the current status of HPV vaccines as well as the most common associated factors that might interfere on establishment of strategies that could control the HPV infections and the

Penile malignancies are thought to arise from the accumulation of multiple mutations that may occur as consequence of progressive genetic instability. This intricate process of genetic instability may be caused by environmental factors, such as history of intense smoking, penile tears, phimosis, and poor genital hygienic habits. (Chaux & Cubilla 2012). In addition, a recent study conducted by Chaux et al. (2011) also described the poor education, penile chronic inflammation, genital warts and Human papillomavirus (related to number of sexual partners

There is a worldwide geographic difference in occurrence of penile malignancies that could be caused by differences in socio-economic status, cultural and religious conditions (Bleeker et al. 2009, Chaux & Cubilla, 2012). The higher incidences are frequent in tropical or subtropical regions of Latin America, Asia and Africa but have uncommon incidence rates in Europe, Japan, USA and Israel (Cubilla 2009). Recently, were reported higher incidence rates in underdeveloped regions such as Africa, South America, and Asia (2-4/100000 inhabitants) as compared with North America (United States) and Europe (0.3-1/100000 inhabitants) (Chaux & Cubilla, 2012). Pow-Sang et al. (2010) also described the penile malignancies prevalence rate among different populations. Prevalence rates in developed countries as Israel (0.1/100 000) and United States (0.3-1.8/100 000) and interestingly, compared with underdeveloped coun‐ tries such as Uganda (2.8/100 000) and Brazil (1.5-3.7/100 000). Once again confirming the

Squamous cell Carcinoma represents vast majority of histological subtype of primary penile malignancies with heterogeneous features due to differences in morphology pathogenesis and prognosis (Hakenberg & Protzel, 2012; Stankiewicz et al., 2012; Syed et al., 2012). Knowledge

development of penile carcinoma associated to this infection.

during lifetime) as environmental factors to malignant transformation.

disease geographical difference and the influence of country development.

cell cycle controls.

**2. Penile cancer**

The anatomy of the penis is complex and has important implications to define predictive risk model and delineate the prognostic factors (Chaux & Cubilla, 2012). The same authors described 3 anatomical compartments in the penis (Glans, Foreskin and Coronal Sulcus) where the malignant neoplasms may be originated (Fig 1). However, the penile malignant neoplasms have a predilection to originate first on the Gland followed by Foreskin inner mucosa and lastly the Coronal Sulcus is rarely affected by neoplastic entity.

**Figure 1.** Paraurethral longitudinal section presenting anatomical levels of the Penis. **CC:** Corpus Cavernosum; **CS:** Cor‐ pus Spongiosum; **LP:** Lamina Propria; **SF:** Skim of the Foreskim and **TA:** Tunica Albuginea. (Adapted from Chaux & Cu‐ billa 2012).

Recently, Hernandez et al. (2008) performed an epidemiological study with 4967 United States men with the diagnosis of penile squamous cell carcinoma. Thirty four percent of patients (1712) presented neoplasms arising in gland, 13.2% in prepuce, 5.3% in penis shaft, 4.5%, in overlapping of penis, and 42,5% in unspecified site. Lesions generally initiate on the glans and slowly extend to involve completely the glans and shaft of the penis. During the neoplasm progress Buck's fascia act as a natural barrier to local tumor invasion defending the corporal bodies from tumoral expansion (Pow-Sang et al. 2010). This assessment is schematically illustrated in figure 2.

The anatomy of the penis presents a pivotal role in tumor invasion and prognosis of cancer. Moreover, the TNM staging system is based, at least partially, on the commitment of these anatomical levels (Velazquez et al. 2010). The glans can be divided in 4 levels: squamous epithelium, lamina propria, corpus spongiosum, and corpus cavernosum (corpus spongio‐ sum, and corpus cavernosum are subdivided by the tunica albugínea). Anatomical levels in the foreskin, like in glans, are divided in squamous epithelium, lamina propria, dartos muscle, and outer skin (Chaux & Cubilla, 2012).

**Figure 2.** Natural history of penile cancer (Adapted from Pow-Sang et al. 2010).

A previous study suggests that different tumor histological features could be based on anatomical site. This hypothesis is sustained by the histological differences among the urethral segments and their corresponding neoplasms (Velasquez et al. 2005). As discussed previously, Squamous Cell Carcinoma of Usual Type (SCCUT) is the most frequent histopathologic diagnoses in penile malignancies (Chaux & Cubilla, 2012; Stankiewicz et al. 2012) affecting around 48%-65% of all type of penile carcinoma. Additionally, Epstein et al. (2011) reported 11 more subtypes of SCCUT (Table 1). Macroscopic features of SCCUT range from endophytic to irregular exophytic masses, presenting white-to-gray coloration. However, the reddish pigmentation also can be observed (Chaux et al. 2010). Microscopilly, the SCCUT is similar to oral, vulvar and cervical Squamous Cell Carcinomas (Cubilla et al. 2001). SCCUT may vary from well-differentiated tumors to anaplastic entities. Other presented feature is keratiniza‐ tion, ranging from highly keratinized, presented in well-differentiated tumors, until scarce or minimal keratinization observed in anaplastic neoplasm. Chaux et al. (2010)

Lopes et al. (2002) performed a study that aimed to investigate the p53 in Brazilian patients with PSCC to establish a new prognostic factor for lymph node metastasis and its possible influence on prognosis. This study observed the nodal stage as a factor that influenced survival (independent risk factors) in the univariate and multivariate analyses. Gunia et al. (2012) shown that p16INK4a is a good prognostic marker for penile squamous cell carcinomas, surpassing the prognostic impact of histologically confirmed koilocytosis. In their study, p16INK4a expression predicted better cancer specific survival rates. Furthermore, p16INK4a can be useful in differentiate subtypes of PSCC. According Chaux & Cubilla (2012), warty carcinomas tend to be p16INK4a positive, whereas giant condylomas and papillary and

**Table 1.** Classification of Squamous Cell Carcinomas (SCCs) of the penis (Adapted from Chaux & Cubilla 2012)

**Classification of SCCs of the penis**

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**Subtype Frequency (%)**

Usual SCC 48-65 Basaloid carcinoma 4-10 Warty carcinoma 7-10 Verrucous carcinoma 3-8 Papillary carcinoma 5-15 Sarcomatoid carcinoma 1-3 Mixed carcinomas 9-10 Adenosquamous carcinoma 1-2 Pseudohyperplastic carcinoma <1 Carcinoma cuniculatum <1 Pseudoglandular carcinoma <1 Warty-basaloid carcinoma 9-14

Medical record analysis of 145 men with penile squamous cell carcinomas was performed to identify prognostic factors for lymph node involvement (Lopes et al. (1996)). The authors found that lymph node metastasis presents correlation with tumor thickness, lymphatic and vascular embolization. Interestingly, univariate analysis did not reached statistically significant values to pathologic stage of primary tumor, clinical lymph node stage (cN), and histological grade. However, histological grade may be considered an important prognostic factor in penile squamous cell carcinoma. In accord with Cubilla AL. 2009 these prognostic factors are

Currently, different methods are employed to grade Penile Squamous Cell Carcinomas. For instance, Akhter et al. (2011) uses the Broder's system as histological grade system in Squamous Cell Carcinoma. In the Broder's system Penile Squamous Cell Carcinomas is stratified in 4 grades levels based only in differentiation of the cells: Grade I (well differentiated) presenting <25% undifferentiated cells; Grade II (Moderately differentiated) presenting <50% undiffer‐

predictive to the nodal spread, metastasis and tumoral dissemination.

verrucous carcinomas are consistently negative.

Evidences from pertinent literature indicate the involvement of groin lymph nodes as the most relevant and unfavorable prognostic factors predicting cancer-specific survival in patients with penile squamous cell carcinoma. Numerically, in the same review, the 5-year cancerspecific survival rate for those presenting cN0 tumors were between 75% and 93%, compared with a 5-year cancer-specific survival rate for those presenting cN3 tumors ranging between 20% and 34%. There is a substantial decrease in the survival rates with N progression (Novara et al. 2007).

**Table 1.** Classification of Squamous Cell Carcinomas (SCCs) of the penis (Adapted from Chaux & Cubilla 2012)

A previous study suggests that different tumor histological features could be based on anatomical site. This hypothesis is sustained by the histological differences among the urethral segments and their corresponding neoplasms (Velasquez et al. 2005). As discussed previously, Squamous Cell Carcinoma of Usual Type (SCCUT) is the most frequent histopathologic diagnoses in penile malignancies (Chaux & Cubilla, 2012; Stankiewicz et al. 2012) affecting around 48%-65% of all type of penile carcinoma. Additionally, Epstein et al. (2011) reported 11 more subtypes of SCCUT (Table 1). Macroscopic features of SCCUT range from endophytic to irregular exophytic masses, presenting white-to-gray coloration. However, the reddish pigmentation also can be observed (Chaux et al. 2010). Microscopilly, the SCCUT is similar to oral, vulvar and cervical Squamous Cell Carcinomas (Cubilla et al. 2001). SCCUT may vary from well-differentiated tumors to anaplastic entities. Other presented feature is keratiniza‐ tion, ranging from highly keratinized, presented in well-differentiated tumors, until scarce or

224 Human Papillomavirus and Related Diseases – From Bench to Bedside A Diagnostic and Preventive Perspective

Evidences from pertinent literature indicate the involvement of groin lymph nodes as the most relevant and unfavorable prognostic factors predicting cancer-specific survival in patients with penile squamous cell carcinoma. Numerically, in the same review, the 5-year cancerspecific survival rate for those presenting cN0 tumors were between 75% and 93%, compared with a 5-year cancer-specific survival rate for those presenting cN3 tumors ranging between 20% and 34%. There is a substantial decrease in the survival rates with N progression (Novara

minimal keratinization observed in anaplastic neoplasm. Chaux et al. (2010)

**Figure 2.** Natural history of penile cancer (Adapted from Pow-Sang et al. 2010).

et al. 2007).

Lopes et al. (2002) performed a study that aimed to investigate the p53 in Brazilian patients with PSCC to establish a new prognostic factor for lymph node metastasis and its possible influence on prognosis. This study observed the nodal stage as a factor that influenced survival (independent risk factors) in the univariate and multivariate analyses. Gunia et al. (2012) shown that p16INK4a is a good prognostic marker for penile squamous cell carcinomas, surpassing the prognostic impact of histologically confirmed koilocytosis. In their study, p16INK4a expression predicted better cancer specific survival rates. Furthermore, p16INK4a can be useful in differentiate subtypes of PSCC. According Chaux & Cubilla (2012), warty carcinomas tend to be p16INK4a positive, whereas giant condylomas and papillary and verrucous carcinomas are consistently negative.

Medical record analysis of 145 men with penile squamous cell carcinomas was performed to identify prognostic factors for lymph node involvement (Lopes et al. (1996)). The authors found that lymph node metastasis presents correlation with tumor thickness, lymphatic and vascular embolization. Interestingly, univariate analysis did not reached statistically significant values to pathologic stage of primary tumor, clinical lymph node stage (cN), and histological grade. However, histological grade may be considered an important prognostic factor in penile squamous cell carcinoma. In accord with Cubilla AL. 2009 these prognostic factors are predictive to the nodal spread, metastasis and tumoral dissemination.

Currently, different methods are employed to grade Penile Squamous Cell Carcinomas. For instance, Akhter et al. (2011) uses the Broder's system as histological grade system in Squamous Cell Carcinoma. In the Broder's system Penile Squamous Cell Carcinomas is stratified in 4 grades levels based only in differentiation of the cells: Grade I (well differentiated) presenting <25% undifferentiated cells; Grade II (Moderately differentiated) presenting <50% undiffer‐ entiated cells; grade III (Poorly differentiated) presenting <75% undifferentiated cells and grade IV (Anaplastic/Pleomorphic) >75% undifferentiated cells. Cell anaplasia degrees are also pointed as common approach to determine Penile Squamous Cell Carcinomas grading (Mikuz et al. 2004; Slaton et al. 2001), absence of anaplasia (well differentiated cells), grade 1; grade 2, moderately differentiated (<50% anaplastic cells); and grade 3, poorly differentiated (>50% anaplastic cells). Cubilla et al. (2009) reported a method to grade Penile Squamous Cell Carcinomas. Carcinomas with a minimal deviation from normal/hyperplastic morphology of squamous epithelium were considered Grade 1 (extremely well-differentiated). Grade 3 are tumors showing any proportion of anaplastic cells, identified as solid sheets or irregular small aggregates, cords or nests of cells with little or no keratinization, high nuclear cytoplasmic ratio, thick nuclear membrane, nuclear pleomorphism, clumped chromatin, prominent nucleoli and numerous mitosis. Grade 2 is composed by remainder tumors. Grading both extremes of the spectrum is simple and reproducible.

established. Specifically, more than a dozen protein-protein interactions between E6 and cellular proteins have been shown (Villa et al., 2002). Taken into a carcinogenic point of view, E6 and E7 ORF are considered to play the most important roles, encoding for oncoproteins that allow viral replication and the immortalization and transformation of the epithelial cell

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Proving the importance of p53 and pRb in cell cycle progression, the repression of HPV 16 E6 and E7 expression by dual shRNA transfection has been shown to be capable of restoring the p53 and pRb tumor suppressor pathways and activating apoptosis (Psyrri et al., 2009, Rampias et al., 2009). Thus, the demonstration of this tumor suppressor inactivation by the E6 and E7 HPV oncoproteins has provided a basic explanation for how the high-risk HPV types exert their oncogenic effects on cervical cells, and this explanation are under investigation to be related with other sites of HPV-infection. This is particularly important in penile cancerassociated HPV infection, whereas HPV16 seems to develop a pivotal role, and accounts for

Different from other viruses, HPV does not infect or replicate in antigen-presenting cells (APCs) of the epithelium nor induce cell lysis, which is a key escape mechanism to avoid that APC recognize and produce antigens derived from the virion, and alerts immune system. About more than 50% of infections present seroconversion in the patients, but the production of antibodies usually occurs only months after the initial infection (Vidal & Gillison, 2008). The life cycle of papillomaviruses is closely tied to the epithelial differentiation process. Infection occurs exclusively in squamous epithelial cells with a preference for the keratinocyte stem cell as the initial target of HPV infection, which will allow the maintenance of viral replication (Vidal & Gillison, 2008). The route of entry for HPV infection is microtraumas or small wounds in the skin or mucosal surface, which are particularly important in penile HPV-infection. These breaks in the epithelial surface allow the virus to access and persist in the nuclei of infected basal layer cells of the epithelium. Until now, no single receptor has been definitively identified and established as being responsible for HPV entry, although is believed that receptors closely related to wound healing might be preferential targets for HPV infection, such as α6 integrin

As most viruses, HPV uses the host cell DNA machinery to maintain the production of viral progeny. This mechanism of viral-induced cell growth is very well known and is analogous to other viruses that disrupt the control of cell growth (Hebner & Laimins, 2006). Following cell division, as the basal cells divide into squamous epithelial cells, HPV establishes its DNA genome in the host cell nuclei, replicates and reaches a high copy number. Infected cells then leave the basal layer, migrate toward the suprabasal regions and begin to differentiate. In the basal layer phase, the HPV genome is maintained at a low copy number, providing a type of stock of viral DNA for further use in cell divisions. At the same time, 'early' viral genes (E5, E6 and E7) are expressed, resulting in enhanced proliferation of the infected cells and their

that host the HPV DNA (Doorbar et al., 1991).

more than 60% of HPV-related tumors.

**4. HPV impact in squamous cell homeostasis**

and glycosamioglycan heparin (Vidal & Gillison, 2008).

In summary, Penile Squamous Cell Carcinomas represents an aggressive locoregional malignancy with dissemination process may occurring 20-40% (Guimarães et al. 2009), a lethal disorder that often presents after significant delay (Barocas & Chang 2010). Usually, death occurs within 2 years after initial diagnosis (Guimarães et al. 2009). Accurate evaluation of clinical stage, anatomical site, regional lymph nodes, and metastatic disease presents a pivotal role in treatment planning to predict the survival outcome (Barocas & Chang, 2010).


**Abstract**

Xenotransplantation, the transfer of cells, tissues or organs between species, has the potential to overcome the critical need for organs to treat patients. One major barrier in the widespread application of xenotransplantation in the clinic is the overwhelming rejection response that occurs when non-human organs encounter the human immune system. Recent progress in developing new and better genome engineering tools now allows the genetic engineering of genes and pathways in non-human animals to overcome the human rejection response and provide an unlimited supply of rejection-free organs. In this review, the benefits and drawbacks of various genome engineering protocols, and examples of their application in xenotransplantation, are discussed.

**Keywords:** xenotransplantation, xenoantigen, transgene, gene targeting, gene editing, homologous recombination


**7. Benefits of exercise**

Regular PE in patients with CKD is associated with a myriad of health benefits, including physiological, psychological, and functional benefits.

## **7.1 Physiological benefits**

Regular PE reduces CV mortality, hypertensive medication use, inflammatory markers (C-reactive protein); prevents muscle wasting; improves toxin removal by dialysis, exercise capacity, blood pressure control, lipid profile (increased HDL-C and reduced TG), haematocrit (prior to erythropoietin therapy), glycaemic control, serum albumin, nutritional summarise.

#### **7.2 Psychological benefits**

There is a link between regular physical activity and a better psychological profile (lower stress/anxiety/hostility/depression and increased engagement in pleasurable activities), perception of general and mental health, physical functioning, and vitality. Subjective weariness symptoms are reduced, as is the impression of physical pain, all at the same time [75–77].

#### **7.3 Functional benefits**

Regular physical exercise improves muscle strength, 6-minute walk distance, gait speed, sit-to-stand time, balance (which reduces the chance of falling), independence, and HRQoL [75–77].


**2. Classification and pathogenesis of DLBCL**

Non-Hodgkin's lymphoma (NHL) classification is based on pathohistological findings. The history of classification of lymphoid neoplasms begins in the 1940s. Based on the advancement in knowledge, the work classification and Kiel have replaced the revised European-American Classification of Lymphomas, and it is a step ahead of the classification given by the WHO [8].

Classifications are an essential part of modern medicine, offering a consensus on the terminology and disease definitions to be used interdisciplinary both in research and clinical practice. Evolution of lymphoma classification includes numerous attempts from descriptive schemes, relying on morphology to strictly clinically oriented stratifications proposed by hematologists, usually without significant international acceptance. The classification of the WHO (Swerdlow et al. 2008) differs from the previous classifications in defining the real entities of the disease through multiple divisions. The new entities defined in this release include extended immunophenotypes and some gene information relevant to the classification. Panel antibody was selected based on morphological differential diagnosis and tumor location. DLBCL is a diffuse proliferation of large neoplastic B lymphoid cells of the same size or up to two times than the normal lymphocyte or extremely large as in macrophages. The origin of these cells are B cells of the germinating center and postgerminant cells [8]. DLBCL can occur in the form of nodal and extranodal diseases. Approximately 40% of patients are present with extranodal disease. Any tissue organ can be the primary site of DLBCL, but the gastrointestinal tract is the most common site [33].

Tumor cells are B large transformed lymphocytes, but the morphological picture of DLBCL is varied, and the predominant morphological cell type can distinguish six subtypes of DLBCL: centroblastic, immunoblastic, T lymphocyte/histiocyte rich, anaplastic, plasmablastic, and DLBCL with cells similar to Reed-Sternberg cells and expression of anaplastic lymphoma kinase (ALK) protein. Tumor cells are positive, as defined by the disease, on pan-B markers CD19, CD20, CD22, and CD79a, and the positivity of intracytoplasmic or surface immunoglobulin (Ig) markers is distinctly expressed as CD5, CD10, CD30, and CD45 markers. Numerous chromosomal abnormalities have been detected with DLBCL, but no specific diagnostic changes have been isolated. The most frequent changes are breaks of regions 14q32, 3p21, 3q27, 22q12, 1q25, and 18q21 and additional chromosomes 7,9,12 and 4q31-35,13p13-14 and 17p11-13,8,9. In 11–23% of cases, DLBCL with advanced clinical stage of the disease revealed nine regions of chromosomal amplification and isolated amplified genes with a potential role in the formation of lymphoma. The clinical significance of each individual change has not been established, but some changes are associated with advanced clinical stage or poorer response to therapy. The abovementioned chromosomal abnormalities are generally not found as individual changes. The accumulation of a greater number of abnormalities indicates the progression of genetic changes, which play an important role in the pathogenesis of DLBCL [8, 33].

**83**

*B Cell Lymphomagenesis*

*DOI: http://dx.doi.org/10.5772/intechopen.87241*

T-cell/histiocyte-rich large B-cell lymphoma Primary DLBCL of the central nervous system

DLBCL associated with chronic inflammation

Primary mediastinal (thymic) large B-cell lymphoma

Germinal center B cell-like (GCB) Activated B-cell-like (ACB)

Primary cutaneous DLBCL, leg type

EBV mucocutataneous ulcer

Lymphomatoid granulomatosis

Burkitt lymphoma

Hodgkin lymphoma

**Table 1.**

Intravascular large B-cell lymphoma ALK+ large B-cell lymphoma Plasmablastic lymphoma Primary effusion lymphoma

Diffuse large B-cell lymphoma (DLBCL), not otherwise specified (NOS)

Epstein–Barr virus (EBV) positive DLBCL, not otherwise specified (NOS)

Human herpesvirus 8 (HHV8)+ DLBCL, not otherwise specified (NOS)

High-grade B-cell lymphoma, with MYC and BCL2 and/or BCL6 translocations

*The 2016 update of WHO classification of DLBCL: subtypes and related entities [8].*

B-cell lymphoma, unclassifiable, with features intermediate between DLBCL and classical

**4. Prognostic markers in DLBCL**

Burkitt-like lymphoma with 11q aberration

High-grade B-cell lymphoma, NOS

**4.1 Gene expression profile (GEP)**

clinical course is completely different [50–52].

Clinical and morphological differences in DLBCL suggest the biological significance of subtypes, which may help in adapting the therapy to be defined. The development of DNA microarray techniques provided the ability to identify gene expression of the tumor with a new molecular entity and a molecular predictor essential for survival. DNA microarray can analyze thousands of sites of previously identified genes. DLBCL has been identified by several tumor markers associated with unfavorable outcome after therapy and which can be linked to tumor cells and normal B lymphocytes. Phenotypic cells resemble normal B lymphocytes, but the

Gene expression profile has shown that DLBCL can be divided into two molecular entities that differ in the gene expression profile. Hans et al. have identified two subgroups of DLBCL [34, 52]. One subgroup was called germinal center B-cell-like (GCB) and has a characteristic gene expression of normal GC B cells, in about 50% of cases, and the second group was called activated B-cell-like (ABC). Type-3 group is defined as nonclassified cases. According to the GEP classification, the observed prognosis distinguishes two major subtypes after the chemotherapy. Patients with GCB lymphoma have a more favorable outcome in 5-year survival than patients in the ABC group (68% vs. 24%) [52]. The distinction between GCB and ABC

#### **3. The WHO classification which was revised in 2016**

See **Table 1**.

*Normal and Malignant B-Cell*

**2. Classification and pathogenesis of DLBCL**

step ahead of the classification given by the WHO [8].

each type of lymphoma. *BCL1* translocation results in disturbed cell cycle regulation. *BCL2* and API2-MALT1 have an antiapoptotic function [49]. API2-apoptosis inhibitor 2-MALT1 (mucosa-associated lymphoid tissue lymphoma translocation gene 1)-induced NF-kappaB activation may contribute to antiapoptotic action probably through NF-kappaB-mediated upregulation of apoptotic inhibitor genes [49].

Non-Hodgkin's lymphoma (NHL) classification is based on pathohistological findings. The history of classification of lymphoid neoplasms begins in the 1940s. Based on the advancement in knowledge, the work classification and Kiel have replaced the revised European-American Classification of Lymphomas, and it is a

Classifications are an essential part of modern medicine, offering a consensus on the terminology and disease definitions to be used interdisciplinary both in research and clinical practice. Evolution of lymphoma classification includes numerous attempts from descriptive schemes, relying on morphology to strictly clinically oriented stratifications proposed by hematologists, usually without significant international acceptance. The classification of the WHO (Swerdlow et al. 2008) differs from the previous classifications in defining the real entities of the disease through multiple divisions. The new entities defined in this release include extended immunophenotypes and some gene information relevant to the classification. Panel antibody was selected based on morphological differential diagnosis and tumor location. DLBCL is a diffuse proliferation of large neoplastic B lymphoid cells of the same size or up to two times than the normal lymphocyte or extremely large as in macrophages. The origin of these cells are B cells of the germinating center and postgerminant cells [8]. DLBCL can occur in the form of nodal and extranodal diseases. Approximately 40% of patients are present with extranodal disease. Any tissue organ can be the primary

site of DLBCL, but the gastrointestinal tract is the most common site [33].

**3. The WHO classification which was revised in 2016**

Tumor cells are B large transformed lymphocytes, but the morphological picture of DLBCL is varied, and the predominant morphological cell type can distinguish six subtypes of DLBCL: centroblastic, immunoblastic, T lymphocyte/histiocyte rich, anaplastic, plasmablastic, and DLBCL with cells similar to Reed-Sternberg cells and expression of anaplastic lymphoma kinase (ALK) protein. Tumor cells are positive, as defined by the disease, on pan-B markers CD19, CD20, CD22, and CD79a, and the positivity of intracytoplasmic or surface immunoglobulin (Ig) markers is distinctly expressed as CD5, CD10, CD30, and CD45 markers. Numerous chromosomal abnormalities have been detected with DLBCL, but no specific diagnostic changes have been isolated. The most frequent changes are breaks of regions 14q32, 3p21, 3q27, 22q12, 1q25, and 18q21 and additional chromosomes 7,9,12 and 4q31-35,13p13-14 and 17p11-13,8,9. In 11–23% of cases, DLBCL with advanced clinical stage of the disease revealed nine regions of chromosomal amplification and isolated amplified genes with a potential role in the formation of lymphoma. The clinical significance of each individual change has not been established, but some changes are associated with advanced clinical stage or poorer response to therapy. The abovementioned chromosomal abnormalities are generally not found as individual changes. The accumulation of a greater number of abnormalities indicates the progression of genetic changes, which play an important role in the pathogenesis of DLBCL [8, 33].

**82**

See **Table 1**.

**Table 1.**

*The 2016 update of WHO classification of DLBCL: subtypes and related entities [8].*


*3.2. Resin-Assisted Capture (RAC) of Oxidized Proteins (OX) Coupled to Mass Spectrometry (OX–RAC) Analysis of E. histolytica Trophozoites Exposed to L. acidophilus*

In order to explore the amebicidal properties of *L. acidophilus*, we used OX–RAC to measure the levels of oxidized proteins (OXs) in *E. histolytica* trophozoites exposed to *L. acidophilus*. In absence of DTT treatment, OXs are not expected to bind to the thiopropyl resin [38]. We observed that the level of OXs in *E. histolytica* trophozoites exposed to heatkilled *L. acidophilus* culture is very low (Figure 2A). These results indicate that heat-killed culture of *L. acidophilus* do not trigger the formation of OXs in *E. histolytica* trophozoites. In contrast, a strong level of OXs was detected in *E. histolytica* trophozoites exposed to live *L. acidophilus* culture (Figure 2A). The addition of catalase during the interaction of *E. histolytica* trophozoites with *L. acidophilus* strongly inhibits the formation of OXs in the parasite, which confirms that the formation of OXs in the parasite is mediated by H2O2 produced by *L. acidophilus* (Figure 2B). These results indicate that the formation of OXs is triggered by H2O2 produced by *L. acidophilus*.

**Figure 2.** Detection of OXs by resin-assisted capture (OX–RAC) analysis of *E. histolytica.* Note: *E. histolytica* trophozoites were incubated with live *L. acidophilus* (L.a) or with heat-killed *L. acidophilus* (DN L.a) (**A**), with/without catalase (Cat.) (50 μg/mL) (**B**) for 2 h at 37 ◦C. Total protein lysate was prepared by lysing the trophozoites with 1% Igepal in PBS. The oxidized proteins in the cell lysates were subjected to RAC in the presence of 10 mM DTT (+DTT) or the absence of DTT (−DTT). The protein was resolved on a 12% SDS-PAGE and stained with silver stain.

The intensity of the protein bands were quantified by densitometry using Image J software [39]. The intensity of the OX-protein bands obtained in the presence of DTT in *E. histolytica* trophozoites incubated with live *L. acidophilus* was arbitrarily set to 1. It is important to note that the data presented in Figure 2A,B were obtained at two different times, and that the silver staining development time was different in each case.

Using MS, we identified 997 OXs in *E. histolytica* trophozoites incubated with *L. acidophilus* (Table S1), which were classified using PANTHER. The most abundant OX families belong to metabolite interconversion enzyme (PC00262), such as protein arginine N-methyltransferase (EHI\_158560), the galactose-specific adhesin 170kD subunit (EHI\_042370), and thioredoxin (EHI\_004490) (Figure 3A). *E. histolytica* lacks glutathione, so it relies mainly on thiol for its defense against OS [40]. Thioredoxin (TRX)/thioredoxin reductase (TRXR) also contributes to redox signaling in *E. histolytica* trophozoites as well as oxidative stress responses [41]. This ubiquitous mechanism of defense is present in many parasites, including *Schistosoma mansoni*, *Plasmodium falciparum*, *Giardia lamblia*, and *Trichomonas vaginalis* [41]. TRXs are small redox proteins of around 12 kD, which act as radical scavengers. In their active site, two cysteine residues are involved in the antioxidant system. The oxidation of these cysteine residues produces disulfide bonds, which will be reduced by TRXR. The presence of TRXs as OXs in *E. histolytica* exposed to *L. acidophilus* strongly suggests that the parasite is actively responding to H2O2 released by the bacteria.

The other abundant OX family belongs to the protein modifying enzyme (PC00260) such as cysteine proteinase CP5 (EHI\_168240), serine/threonine-protein phosphatase (EHI\_031240), or E3 ubiquitin-protein ligase (EHI\_050540) and the protein-binding activity modulator (PC00095) such as AIG1 family protein (EHI\_176700), inhibitors of serine proteinase domain-containing protein (SERPIN) (EHI\_119330), and the Rho family GTPase (EHI\_070730) (Figure 3A).

SERPINs control a broad range of biological processes, including pathogen evasion of the host defense system. Cathepsin G, a pro-inflammatory enzyme released by activated neutrophils, is inhibited by serpins [42]. *E. histolytica* expresses a SERPIN that interacts with human neutrophil cathepsin G [43]. In this work, we showed that EhSERPIN is one of the OXs present in *E. histolytica* exposed to *L. acidophilus*. Studies have suggested that SERPINs are redox-regulated by oxidation of cysteine residues in the reactive site loop of these enzymes or its vicinity [44–46]. The presence of carbamidomethylated cysteine residues in the vicinity of the reactive site loop of EhSERPIN (Table S2) [43] suggests that EhSERPIN is also redox-regulated. The effect of oxidation on EhSERPIN activity has yet to be determined.

A functional motility is critical to the survival of *E. histolytica* in order to both dislodge and phagocytose host cells as well as transport virulence factors intracellularly [47]. Rho GTPases play a critical role in the regulation of motility and phagocytic activity of *E. histolytica* [48]. There are several Rho GTPases present in the parasite, and we identified six of them (EHI\_126310, EHI\_013260, EHI\_197840, EHI\_029020, EHI\_129750, and EHI\_070730) as OXs. EhRho1 (EHI\_029020) regulates phagocytosis by regulating actin polymerization [49]. Numerous studies have shown that ROS regulate Rho GTPases activity [50]. Many Rho family GTPases contain a cysteine-containing motif (GXXXXGK[S/T]C) at their N-terminal, which is located directly adjacent to the phosphoryl-binding loop. Oxidation of the cysteine residue in this motif affects the nucleotide binding properties of these Rho GTPases [50]. According to the MS analysis of OXs (Table S2), this cysteine residue in the active site is not carbamidomethylated. Instead, we found that cysteine residues located at the C-terminal of these Rho GTPases are carbamidomethylated (Table S2). An ubiquitination region is present in the C-terminal region of many Rho GTPases that may regulate their stability [51]. In light of this information, it is tempting to speculate that the stability of these Rho GTPases is redox-dependent. An example of such regulation occurring in human endothelial cells is described here [52].

**Figure 3.** Protein analysis through evolutionary relationships (PANTHER) analysis of OXs in *E. histolytica* incubated with *L. acidophilus.* Note: (**A**) PANTHER sequence classification of the OXs identified in *E. histolytica* trophozoites co-incubated with *L. acidophilus*. (**B**) PANTHER statistical overrepresentation test of the OXs identified in *E. histolytica* trophozoites incubated with *L. acidophilus*.

Of the OXs in *E. histolytica* trophozoites incubated with *L. acidophilus* (Table S2), oxidoreductase (PC00176) and dehydrogenase (PC00092), such as glyceraldehyde-3-phosphate dehydrogenase (EHI\_008200), NAD(FAD)-dependent dehydrogenase (EHI\_099700), and pyruvate: ferredoxin oxidoreductase (EHI\_051060), are significantly enriched according to the PANTHER statistical overrepresentation test (Figure 3B). Pyruvate: ferredoxin oxidoreductase (EHI\_051060) is a Fe–S enzyme that catalyzes the oxidative decarboxylation of pyruvate [53]. This protein has also been identified as an OX in trophozoites exposed to

H2O2 [6], metronidazole, or auranofin [54]. In an oxidatively stressed parasite, pyruvate: ferredoxin oxidoreductase becomes strongly inhibited, resulting in an accumulation of pyruvate, which limits ATP production and causes parasite death [55]. Several cysteine residues present within the [4Fe–4S] clusters of close to them are carbamidomethylated suggesting that they are oxidized (Table S2). Destabilization of the Fe–S clusters integrity via oxidation of these cysteine residues in the parasite exposed to *L. acidophilus* will more certainly inactivate the enzyme and consequently contribute to the parasite death.

Other OXs, which are significantly enriched according to the PANTHER statistical overrepresentation test, include vesicle coat protein (PC00235), such as GOLD domaincontaining protein (EHI\_023070), beta2-COP (EHI\_088220) and coatomer subunit gamma (EHI\_040700) and protease (PC00190), such as EhCP-a1 (EHI\_074180) and EhCP-a4 (EHI\_ 050570) (Figure 3B).


**3. Discussion**

Numerous variations occur in the chloroplast genomes of parasitic plants. However, only a small number of plants within Santalales have been studied. In this study, the complete chloroplast genomes of *M. cochinchinensis*, *M. tricolor* and *M. bibracteolatus* from Santalales were assembled, annotated and analyzed. Compared with the chloroplast genomes of the model plant *Nicotiana tabacum*, all the *ndh* genes of the chloroplast genomes were lost amongst the three species, and the *infA* gene, which codes for a translation initiation factor, was also missing in these species. These cases were similar to those of *T. chinensis* and *T. sutchuenensis* [7]. The *rpl16* and *ycf15* genes were lost in the three species, but they were still present in *T. chinensis* as pseudogenes (Figure 9). However, compared with the results reported by Shin et al. [36], different gene contents of the chloroplast genome were observed in *M. cochinchinensis*. These studies have shown that *M. cochinchinensis* contains the exon 1 fragment of the *ndhB* gene and a fragment of the *infA* gene, whereas the *rpl36* gene is completely lost. However, the *rpl36* gene is still present in the chloroplast genome according to our results. *M. cochinchinensis* has lost the *infA* gene and all *ndh* genes. The number of tRNA genes also differed between the two studies. We annotated 35 tRNA genes, but previous studies only obtained 30 tRNA genes. The evolution of the chloroplast genome in parasitic plants, particularly nonphotosynthetic holoparasites, can lead to significantly reconfigured plastomes [21]. The losses of *ndh* genes are associated with nutritional status or extensive rearrangements of chloroplast structures [37], and they have occurred in the reported chloroplast genomes of parasitic plants [7]. Our study also showed that *ndh* genes were lost in the transformation from autotrophy to heterotrophy [38].

**Figure 9.** Comparison of the chloroplast genome gene content of six parasitic plants and one model plant (*Nicotiana tabacum*). The common existing genes in the complete chloroplast genome of the seven species are not listed. Red boxes indicate each gene present, and green boxes indicate that each gene is considered as a pseudogene. The yellow boxes indicate an absent gene.

The Santalales order consists of a small number of autotrophic species and a large number of parasitic species which are root or aerial (stem) parasites [39]. According to the Engler system, Santalales consists of seven families. We downloaded five families belonging to Santalales, which were available in the National Center for Biotechnology Information (NCBI) at that time, and two

species as outgroups to analyze the phylogenetic relationships of *Macrosolen* in Santalales. The present study showed that Loranthaceae is closely related to Olacaceae, whereas Viscaceae is closely related to Santalaceae and Opiliaceae. These results are similar to those of previous studies [13,14]. All the phylogenetic results strongly support that Loranthaceae and Viscaceae diverged independently from each other.

As folk medicine in China, *M. cochinchinensis*, *M. tricolor* and *M. bibracteolatus* have been used to treat diseases for a long time, and their dried stems and branches with leaves are used as medicinal parts. However, *Macrosolen* species are similar in appearance, especially when they are processed into medicinal slices, thereby causing difficulty in their identification. The identification of parasitic medicinal materials has rarely been reported. Though phytochemical approaches have played an important role in species identification [26], they are inadequate because they are limited to the environment and harvest period. Molecular characterization has shown an improved specificity for plants [23,26]. In our study, mutational hotspots such as the *ycf1* gene, the *ccsA* gene and the *trnF-trnM* intergenic region are potential sites for identification of *Macrosolen* species.


*<IP_ADDRESS> Blastomycosis*

*Sarcoidosis and Granulomatosis - Diagnosis and Management*

*<IP_ADDRESS> Cryptococcosis (European blastomycosis)*

(**Figure 11**).

*<IP_ADDRESS> Coccidiosis*

examination [15].

matosis is more often described in *H. capsulatum* infection [15].

cause the formation of epithelioid cell granulomas; however, necrotizing granulo-

*Cryptococcus neoformans* is ubiquitous; it is found in soil and pigeon excrements. The fungal cell is of 4–7 μm in size, replicates by budding, and is stained with H&E, mucicarmin, and PAS. Cryptococci cause a spectrum of various changes in the lungs. A typical granulomatous reaction presents with confluent non-necrotizing granulomas, many multinucleated giant cells, and a mild inflammatory reaction; giant cells are located mainly outside the granulomas and contain cryptococcal cells

These fungi can also be located inside necrotizing granulomas (cryptococcomas) resembling those with mycobacterial and other types of fungal infections (**Figure 12**). In immunocompromised individuals, cryptococcal cells are found inside the alveoli, in their walls, and in the interstitium, without marked inflamma-

*Coccidia* most often lead to necrotizing granuloma formation; the eosinophilic reaction may be marked or absent; numerous neutrophils may be observed. Like in other infections, granulomas are located peribronchiolar or with destroyed bronchioles. This process is accompanied by the formation of small non-necrotizing granulomas at the periphery. *Coccidia* are usually found in the center of necrotizing granulomas, they consist of large spherical structures (spherules) containing yeast-like structures (endospores). Endospores of various sizes can be located in necrosis or cellular debris, resembling other fungal infections. The detection of spherules and endospores favor the diagnosis of coccidiosis. Like *Histoplasma, Coccidia* do not grow in vitro; thus, the diagnosis can only be made by histological

*Cryptococci cells and multinucleated giant cells in a granuloma. Combined stain of alcian blue and PAS.*

tory reaction; some scattered multinucleated giant cells can be found [9].

**56**

**Figure 11.**

Blastomycosis is a rare disease, and this diagnosis may be suspected when granulomatosis or giant cell lesion in combination with severe acute inflammation is detected. Blastomycosis is characterized by basophilic necrosis rich in cellular debris in contrast to eosinophilic or slightly "dirty" infectious necrotizing granulomas. Blastomycosis is often a bronchiolocentric process. Large, thickwalled, yeast-like *Blastomyces* cells can be detected in H&E and also mucicarmin staining. Active budding is a distinctive feature of the microorganism; nuclear material (multiple nucleoli) can also be found inside the cells, but these signs are not always observed. In this regard, compared to *Histoplasma* and *Cryptococci* cells and *coccidial* endospores, *Blastomyces* cells are larger but still smaller than *coccidial* spherules [15]*.*


**Table 1.**

**107**

*lncRNAs in Hallmarks of Cancer and Clinical Applications*

be promising as biomarkers and potential therapeutic targets.

Leandro Garcia, Erika Zambalde, Carolina Mathias, Jéssica Barazetti,

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

\*Address all correspondence to<EMAIL_ADDRESS>
The authors declare no conflict of interest.

The vast number of studies describing lncRNAs associated with several tumor types and regulating several processes of cancer cells is shown here. The great advance in RNA sequencing technology allows us to identify new molecules and characterized better lncRNAs. From the discovery of these molecules, in the beginning, they appeared to have no important functions; however, today many researches in this area propose that more information about these molecules may help us understand numerous characters of tumor cells that are still unknown. Some lncRNAs are associated with several hallmarks of cancer demonstrating the importance of these molecules in the mechanism of disease, like MALAT and HOTAIR. Other are already utilized as biomarker in prostate cancer like PCA3. Considering the challenges for *in vivo* experimental designs, lncRNAs continue to

*DOI: http://dx.doi.org/10.5772/intechopen.88903*

**4. Conclusion**

**Conflict of interest**

**Author details**

Daniela Gradia and Jaqueline Oliveira\* Federal University of Parana, Curitiba, Brazil

provided the original work is properly cited.

*lncRNAs with potential to be used as biomarkers, in several types of cancer.*


**2. Iodine species in nature**

Iodine plays an integral role in a diverse array of processes. As such, it exists in a variety of forms reflecting either the environment in which it is found or its biological function. Water,

A Review of Spectrophotometric and Chromatographic Methods

**2.4 Food and plants**

and inorganic components (Sheppard et al., 1995; Whitehead, 1984).

and Sample Preparation Procedures for Determination of Iodine in Miscellaneous Matrices 373

mg/kg (Muramatsu, 2004). Retention of iodine in the soil is influenced by a number of factors, including the soil pH, moisture content, porosity and composition of the organic

Geographic differences in topsoil iodine content and irrigation procedures determine food iodine levels. In most diets the mainstay sources of iodine are fish, shellfish, milk and iodinated salt. Food supplements constitute an alternative means of obtaining dietary iodine. Fish contain iodine in similar forms to those found in humans. Perhaps the widest assortment of forms is encountered in different species of seaweed. Brown seaweeds contain mostly I-, however green seaweeds play host to a wide array of organic molecules to which iodine is bound, including numerous proteins and polyphenols. Dietary iodine is obtained from a variety of sources and individual dietary habits contribute to the wide disparity in iodine intake among populations. In 1993, the World Health Organization [WHO] published the first version of the WHO Global Database on Iodine Deficiency with global estimates on the prevalence of iodine deficiency based on total goitre prevalence (TGP), using data from 121 countries. Since the international community and the authorities in most countries where IDD was identified as a public health problem have taken measures to control iodine deficiency, in particular through salt iodization programmes – the WHO recommended strategy to prevent and control IDD (World Health Organization [WHO], 2004). Salt iodization programmes are carried out in more than seventy countries, including the United States of America and Canada. There is a wide variation in the scope of iodine supplement; almost 90% of households in North and South America utilize iodized salt while in Europe and the East Mediterranean regions this figure is less than 50%, with a worldwide figure about 70% (WHO, 2007). WHO, ICCIDD (International Council for the Control of Iodine Deficiency Disorders) and UNICEF (United Nations International Children's Emergency Fund) recommend that the term "iodized" be used to designate the addition of iodine to any substance, regardless of the form. Iodine is commonly added as the I- or IO3- of potassium, calcium or sodium. 70% of salt sold for household use in the U.S.A. is iodized with 100 ppm KI (400 µg iodine per teaspoon) (U.S. Salt Institute, 2007). In Canada all salt must be iodized with 77 ppm KI. Mexico requires 20 ppm levels of iodization. Recommendations for the maximum and minimum levels of iodization of salt are calculated as iodine and determined by local national health authorities in accordance with regional variations in iodine deficiency. In Poland iodine deficiency prophylaxis was first started in 1935. Near the end of the 1940's and 80's, the practice of iodizing table salt was abandoned. A nonobligatory recommendation of iodizing salt took place in 1989. In 1991 the Polish Council for Control of Iodine Deficiency Disorders (PCCIDD) was established and an epidemiological survey performed in 1992-1993, defined Poland as an area with moderated – the seaside region as light – severity of iodine deficiency. In 1996 the production of table salt without the addition of KI was made illegal. An obligatory law was passed, mandating the addition of 30(+/-10) mg of KI per kilogram of salt (Szybiński, 2009). Iodization of salt in Turkey has been mandatory since 1998 and the recommended iodine concentration is 50-70 mg KI/kg or 25- 40 mg KIO3/kg (Gurkan et al., 2004). In Switzerland, iodization of salt was altered three times. Iodization was first introduced in 1922 at 3.75 mg/kg. In 1962 the concentration was doubled and in 1980 it was doubled again giving a present level of 15 mg/kg. Although it's use is voluntary, by 1988, 92% of retail salt and 76% of all salt for human consumption

air, soil and food constitute the most common group of analyzed samples derived from our external environment.

#### **2.1 Water**

In water iodine is predominantly found in the iodide (I-) or iodate (IO3 -) form (Gilfedder et al., 2007; Schwehr & Santschi, 2003). Other forms of iodine species in water include: IO4- (periodate), IO- (hypoiodite), CH3I (methyl iodide), CH2I2 (methyl diiodide), C2H5I (ethyl iodide), C3H7I (propyl iodide), C 4H9I (butyl iodide), and CH2 BrI (methyl iodide bromide) (Hou, 1999, Hou et al., 2009). Organic iodine concentrations may be especially high in fresh water (from rivers, lakes and rain).

#### **2.2 Air**

Generally, the air contains iodine in particulate form, as inorganic gaseous iodine (I2, HIO) and as some forms of organic gaseous iodine (CH3l, CH2l2). High iodine concentrations are found in urban areas due to the combustion of oil and coal. Coastal areas also have high iodine concentration due to the emission of gaseous I2 from algae, seawater and sea spray, which varies with location, season and climate (Hou, 2009; Yoshida & Muramatsu, 1995). There are numerous pathways that may be responsible for transferring I2 from the sea to air. Photochemical oxidation of I from seawater to elemental iodine has been recreated in the laboratory (Miyake & Tsunogai, 1963). Another means of obtaining elemental I2 is through the reaction of I with ozone (O3)(Garland & Curtis, 1981). The greatest source of atmospheric I2 is thought to be from microbial activity within the oceans, through the transformation of I and IO3 into organic CH3I, which has a residence time between 1.1-8 days (Cicerone, 1981). Lovelock et al. (Lovelock et al., 1973) measured the mean atmospheric CH3I concentration above the Atlantic as 1.2 ppt (6.8 ng/m3). Rasmussen et al. (Rasmussen et al., 1982) found the background level of CH3I to vary between 1 and 3 ppt (5.7-17 ng/m3) with measurements near oceans with high biomass productivity to be around 7-22 ppt (40-125 ng/m3).

#### **2.3 Soil**

Strong evidence suggests that atmospheric transport from the oceans is responsible for the deposition of iodine in soil. Iodine exists in various forms in soil and varies largely with respect to its concentration. I- (more mobile form) is believed to be the dominant species in acidic soils whilst IO3- (less mobile) will occur in alkaline soils. In low pH oxidizers, e.g. Fe3+ and Mn4+ convert I - into molecular I2. The activity of reducing bacteria impacts the form of iodine in the soil as well. CH2I2 and other volatile organic complexes of iodine are generated by microbial activity (Johnson, C.C. 2003). Generally, organic bound iodine is more abundant in soil samples. The secondary environment (soil) has high iodine content compared to the primary environment (parent rocks) from which it is derived as a result of weathering. Weathered rocks and soils are richer in iodine than the unweathered bedrocks (Fuge & Johnson, 1986). On average, the igneous rocks contain an average of 0.25 mg/kg of iodine, the sedimentary rocks have 2.3 mg /kg iodine and the metamorphic rocks have 0.81 mg/kg of iodine. Organic matter is the major concentrator of iodine in sedimentary basins (Mani et al., 2007). The highest values were found in soil samples from areas close to the coast, where there is high rainfall, and from areas with high soil organic matter. In one study, the iodine concentration in Japanese soils was found to range from 0.2 mg/kg to 150 mg/kg (Muramatsu, 2004). Retention of iodine in the soil is influenced by a number of factors, including the soil pH, moisture content, porosity and composition of the organic and inorganic components (Sheppard et al., 1995; Whitehead, 1984).


*Proceeding Paper* **The Influence of the Ageing Process on the Mechanical Properties of Cement Mortars with Nano-SiO2 Admixture Initially Subjected to Thermal Treatment †**

**Elzbieta Horszczaruk ˙ 1,\* and Paweł Łukowski 2**

**Abstract:** The research aimed to evaluate the effect of nano-silica (NS) on the strength recovery of the previously heated cement mortars. One hundred twenty cement mortar specimens were prepared with 1% to 5% of NS. The specimens were heated at temperatures ranging from 200 ◦C to 800 ◦C. Half of the samples, after heating and cooling, were mechanically tested. The other half of the specimens were stored for 24 months in 90% relative humidity. The presence of NS fostered partial recovery of the compressive strength of the heated mortars. The cement mortars regained more than 40% of their compressive strength after heating at 400 ◦C and about 25% after heating at 600 ◦C. The strength recovery phenomenon faded away at 800 ◦C.

**Keywords:** cement mortars; compressive strength; high temperature; nano-silica

**Citation:** Horszczaruk, E.; Łukowski, P. The Influence of the Ageing Process on the Mechanical Properties of Cement Mortars with Nano-SiO2 Admixture Initially Subjected to Thermal Treatment. *Mater. Proc.* **2023**, *13*, 20. https://doi.org/10.3390/ materproc2023013020

Academic Editors: Katarzyna Mróz, Tomasz Tracz, Tomasz Zdeb and Izabela Hager

Published: 14 February 2023

**Copyright:** © 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/).


**5. Current application**

The BN material shows excellent chemical, thermal, and mechanical properties, which are utilized in different applications. The industrial application of 2D BN is yet to be realized due to the lack of large-area thin film synthesis techniques. To date, large-area films with uniform thickness are hard to fabricate. The h-BN is known for its wide direct bandgap (~5.9 eV), small lattice mismatch with graphene (1.7%), and high thermal conductivity. The BN nanosheets have been employed in several semiconductors device applications, such as transparent membranes, encapsulation materials, tunneling barriers, and high dielectric materials [81]. For instance, some graphene devices fabricated on the h-BN show very high mobility in order of 60,000 cm2V-1 s-1, which is far greater than III-V devices that exploit 2DEG (two-dimensional electron gas) properties [82].

Interestingly, an ultrathin layer of BN is sandwiched between the graphene layer (C-BN-C), making a field effect tunneling transistor heterostructure. The study revealed that the h-BN nanosheet forms a good tunnel barrier [83]. Ranjan et al. studied dielectric breakdown failure of thin h-BN films. The study found that the breakdown field is 21 MV cm−1 for 3-nm-thick h-BN. The breakdown field suggests that h-BN is more suitable for gate dielectric than high-quality silicon dioxide [84].


**Preface to "Cyclodextrin Chemistry and Toxicology"**

Cyclodextrins (CDs) are cyclic oligosaccharides composed of six (α-CD), seven (β-CD), or eight (γ-CD) D (+) - glucose units linked by α-1,4 bonds. They have found wide application in numerous fields which, thanks to their typical toroidal structure consisting of an apolar internal cavity and a polar external surface, allows the physical inclusion of a wide range of molecules. Cyclodextrins have been the subject of several books, reviews and articles. This testifies to the great interest in these molecules and their derivatives due to the extensive use in various fields, such as biotechnology, chemistry, green chemistry, analytical chemistry, pharmacy, administration of drugs and pharmaceutical excipients, biology, medicine and biomedicine, food and beverage industry, cosmetic formulations/fragrance stabilization, chromatography, catalysis, biotechnology, agrochemistry and the remediation of polluted sites.

This collection of articles, through eleven contributions and three reviews, provides an overview of the applications of cyclodextrins, implements the information regarding the use of cyclodextrins and their inclusion complexes, considering both experimental and theorists approaches and using various scientific and technological tools. Furthermore, the importance of understanding the mechanisms involved in the formation of the complexes and of the resulting advantages, together with their possible integration within different application areas, including those related to data communication, are underlined.

In particular, the published articles have clearly highlighted the dynamism and breadth of the sectors involved, as well as the variety of approaches to the use of CDs: as food supplements and nutraceuticals by studying the impact on the grain dough and on the properties of bread, the effect on the antiplatelet capacity of proteins, development of new formulations with greater solubility, bioavailability and stability, the administration of drugs. The drug concentration plays an important role in the interaction with drug carriers that influence the kinetics of the release process and the toxicological effects. The Molecular Dynamics simulations are useful for evaluating the bioeffects of β-cyclodextrin-based nanosponges on two-dimensional cell cultures, also discussing the study of the cyclodextrin-based nanosystems in targeted cancer therapy. One of the reviews introduces the advances in cyclodextrin (pCD) polymer research, including their synthesis and applications in analytical separation science, materials science and biomedicine. Another review summarizes and highlights the results obtained in the field of cyclodextrin-based contrast agents for medical imaging. Finally, new perspectives are presented in the use of cyclodextrins in textiles, as smart materials for microbial control.

As guest editor of the Special Issue "Cyclodextrin Chemistry and Toxicology", we would like to express our deep appreciation to all the authors whose valuable work has been published in this issue, thus contributing to the success of the Special Issue. Despite the presence on the market of numerous food, cosmetic and pharmaceutical formulations, cyclodextrins continue to attract the interest of both the academic community and the industry and we believe that the rapid evolution of the different sectors considered is moving further in interdisciplinary way in the near future, this will give birth to new discoveries and applications based on innovative scientific insights carrying further technological advances, demonstrating that cyclodextrins, for research, still have a lot of interest.