**3. Results**

#### *3.1. Inhibition of Airway Inflammation of CS-Exposed Airways by YE*

This study examined how YE inhibited CS-evoked inflammation in mouse airways. Exposure of mice to CS increased total leukocyte number in the BALF by ≈1.5-fold (Figure 1A). Surprisingly, the challenge of CS promoted the influx of neutrophils and eosinophils in the BALF, indicating that CS resulted in neutrophilic and eosinophilic inflammation (Figure 1A). Oral administration of ≥25 mg/kg YE reduced CS-induced leukocytosis of neutrophils and eosinophils, which was incomparable to that of control mice (Figure 1A). In addition, YE curtailed the number of lymphocytes and monocytes in the BALF elevated in CS-exposed mice.

It was examined whether CS-stimulated airway inflammation was attenuated in YE-supplemented mice. The tissue level of COX-2 responsible for prostaglandin biosynthesis and inflammation was elevated in CS-exposed mouse lungs (Figure 1B). In addition, the airway tissue levels of iNOS and ICAM-1 directly involved in inflammatory responses were enhanced in CS-challenged mice (Figure 1C,D). However, orally-administrated YE reduced the induction of these inflammatory proteins promoted by exposure of airways to CS (Figure 1B–D).

**Figure 1.** Leukocytes in the bronchoalveolar lavage fluid (BALF; **A**) and inhibition of pulmonary inflammation (**B**–**D**) in cigarette smoke (CS)-challenged mouse lungs treated with dried yeast extracts (YE). Mice were orally administrated with 25–100 mg/kg YE and exposed to CS for 30 min. Cells in BALF were counted using a Hemavet HV950 Multispecies Hematologic Analyzer (**A**). Tissue extracts were subject to Western blot analysis with a primary antibody against COX-2, iNOS, or ICAM-1. β-Actin protein was used as an internal control. The bottom bar graphs represent quantitative results of the upper bands obtained from a densitometer. Values (mean ± SEM, *n* = 3–4) in respective bar graphs not sharing a same small letter indicate a significant difference at *p* < 0.05.

#### *3.2. Blockade of Emphysematous Injury of CS-Challenged Airways by YE*

The histological examination was conducted in the lung tissues stained with H&E. The CS exposure induced the airway wall damage in mice (Figure 2). However, oral administration of 25–100 mg/kg YE highly attenuated the pathological alterations observed in bronchiolar and alveolar tissues of CS-challenged mice. In addition, MMP-12 was highly expressed in mouse airways exposed to CS, evidenced by FITC-immunostaining (Figure 3). In contrast, the treatment with 25–100 mg/kg YE reduced its induction in CS-exposed bronchioles (Figure 2). Moreover, the H&E histological staining revealed that the CS challenge to mice evoked destruction in pulmonary alveoli (Figure 2). Consistently, the MMP-12 induction was markedly enhanced. However, 25–100 mg/kg YE diminished the emphysematous damage in pulmonary alveoli and curtailed the FITC-staining of MMP-12 (Figure 3). Therefore, YE may inhibit emphysema and alveolar cell loss in CS-exposed small airways and alveoli.

**Figure 2.** Blockade of airway destruction by dried yeast extracts (YE) in cigarette smoke (CS)-challenged mouse bronchioles and alveoli. Mice were orally administrated with 25–100 mg/kg YE and exposed to CS for 30 min. Airway tissue sections were stained by using a hematoxylin and eosin (H&E) reagent. Each photograph is representative of four mice. The arrows indicate damaged bronchioles and alveolar air sacs. Scale bars = 100 μm.

#### *3.3. Inhibitory E*ff*ects of YE on CS-Induced Pulmonary Apoptosis and Oxidative Stress*

This study attempted to examine whether YE inhibited emphysematous airway damage through blocking pulmonary apoptosis and oxidative stress induced by CS. Western blot analysis showed that CS diminished the lung tissue level of anti-apoptotic bcl-2 and increased the level of pro-apoptotic bax, consequently elevating the bax/bcl-2 ratio (Figure 4A). Oral treatment of YE reduced the bax/bcl-2 ratio in CS-exposed mouse alveolar tissues. The tumor suppressor p53 is known to directly activate bax and mediate mitochondrial membrane permeabilization and apoptosis [31]. As expected, the activation of p53 was enhanced in CS-loaded lung tissues, which was retarded by supplementing 25–100 mg/kg YE to mice (Figure 4B). In addition, caspase-9 and its downstream executioner caspase-3 responsible for executing cell death were upregulated in lung tissues by exposure to CS (Figure 4C). In contrast, YE highly attenuated the activation of these caspases in lung tissues.

**Figure 4.** Inhibition of apoptotic lung injury and reactive oxygen species (ROS) production by 25–100 mg/kg yeast extracts (YE) in cigarette smoke (CS)-challenged mouse lungs. Tissue extracts were subject to Western blot with a primary antibody against bcl-2, bax, phospho-p53, cleaved caspase-9, or cleaved caspase-3 (**A**–**C**). β-Actin protein was used as an internal control. The bar graphs (mean ± SEM, *n* = 3) represent quantitative results of the upper bands obtained from a densitometer. Values in respective bar graphs not sharing a same small letter indicate a significant difference at *p* < 0.05. Dihydroethidium (DHE) staining showing pulmonary ROS production (**D**). Tissue sections of small airways and alveoli were stained with DHE, and nuclear staining was done with DAPI (blue). Each photograph is representative of four mice. Scale bars=100 μm.

The reciprocal interactions among ROS, airway inflammation, and alveolar cell death play crucial role in the pathogenesis of COPD [32]. This study introduced DHE staining for ROS production in airways exposed to CS. DHE exhibits blue-fluorescence in the cytosol until oxidized, where it intercalates within the cell DNA, with subsequent staining of nuclei as a bright fluorescent red. There was a strong DHE staining in CS-loaded bronchioles and alveoli, indicating marked superoxide

production by CS (Figure 4D). However, oral administration of 25–100 mg/kg YE highly attenuated ROS production in bronchiolar and alveolar tissues of CS-challenged mice.

#### *3.4. Suppressive E*ff*ects of YE on CSE-Loaded Alveolar Apoptotic Injury*

This study further explored how CS evoked alveolar damage in mice and how YE protected alveoli from CS. The treatment of A549 cells with 10–100 μg/mL YE did not cause cytotoxicity for 24 h (Figure 5A). When 5% CSE was applied to alveolar epithelial A549 cells for 24 h, the viability dropped to below 20% (Figure 5B). When 5% CSE-loaded A549 cells were supplemented with ≥10 μg/mL YE for 24 h, the viability was highly boosted (Figure 5B). On the other hand, Hoechst 33258 nuclear staining and TUNEL staining showed that 5% CSE resulted in nuclear condensation and DNA fragmentation of A549 cells in an apoptotic manner (Figure 5C). The apoptotic cell death by CSE was significantly curtailed in YE-added alveolar cells. Accordingly, CS-induced alveolar emphysema may be ascribed to its apoptotic death of alveolar cells.

**Figure 5.** Viability of alveolar epithelial A549 cells and effects of dried yeast extracts (YE) on alveolar apoptosis. Alveolar epithelial cells were incubated in media containing 5% cigarette smoke extract (CSE) in the absence and presence of 10–100 μg/mL YE for up to 24 h. A549 cell viability (mean ± SEM, *n* = 5) was measured by using MTT assay and expressed as percent cell survival relative to untreated controls (**A**,**B**). Values in respective bar graphs not sharing a same small letter indicate a significant difference at *p* < 0.05. Nuclear staining was done with Hoechst 33258 dye for the detection of apoptotic cells (**C**). A transferase dUTP nick end labeling (TUNEL) assay was conducted to detect DNA fragmentation of apoptotic A549 cells and nuclear staining was accomplished with DAPI (**C**). Representative microphotographs were obtained by fluorescence microscopy. Scale bars = 50 μm.

#### *3.5. Inhibition of Airway Inflammation of OVA-Exposed Airways by YE*

This study investigated that YE inhibited allergic airway inflammation evoked by OVA in mouse airways. When mice underwent OVA inhalation, total leukocyte number in BALF was highly elevated by ≈2.5-fold (Figure 6A). The OVA inhalation prompted neutrophilic and eosinophilic inflammation in BALF, while oral administration of 25–100 mg/kg YE reduced OVA-induced leukocytosis of neutrophils and lymphocytes (Figure 6A). In addition, YE diminished eosinophilic inflammation elevated in OVA-exposed mice (Figure 6A).

The tissue levels of inflammatory COX-2 and iNOS were enhanced in OVA inhalation-experienced mouse lungs in a similar manner to the CS challenge (Figure 6B). However, the YE supply attenuated the lung tissue induction of these proteins promoted by OVA (Figure 6B). Accordingly, YE may alleviate OVA inhalation-induced allergic inflammation in airways. In addition, this study examined whether OVA induced pulmonary emphysema in mice. The Cy3-immunostaining revealed that OVA promoted the MMP-12 expression in mouse bronchioles and alveoli (Figure 6C). It should be noted that the MMP-12 induction by OVA inhalation was less than that of CS challenge. Nevertheless, YE curtailed the Cy3-MMP-12 staining in airways, indicating that YE abrogated pulmonary emphysema due to OVA (Figure 6C).

**Figure 6.** Suppressive effects of dried yeast extract (YE) on airway inflammation and induction of airway target proteins in ovalbumin (OVA) inhalation-challenged mouse lungs. OVA-sensitized mice were orally administrated with 25–100 mg/kg YE. Cells in BALF were counted using a Hemavet HV950 Multispecies Hematologic Analyzer (**A**). Lung tissue extracts were subject to Western blot with a primary antibody against COX-2 and iNOS (**B**). β-Actin protein was used as an internal control. The bar graphs (mean ± SEM, *n* = 3) represent quantitative results of the left bands obtained from a densitometer. Values in respective bar graphs not sharing a same small letter indicate a significant difference at *p* < 0.05 Immunohistofluorescence analysis was done in tissues of small airways and alveoli of OA-challenged mice (**C**). The MMP-12 localization was identified as Cy3-red staining in mouse airways exposed to OVA. Nuclear staining was done with DAPI (blue). Each photograph is representative of four mice. Scale bars = 100 μm.

#### *3.6. Blockade of LPS-Triggered Airway Inflammation by YE*

The endotoxin LPS stimulated alveolar inflammation through the induction of COX-2, iNOS, and ICAM-1 in A549 cells (Figure 7A–C). In addition, LPS prompted the secretion of pro-inflammatory cytokines of TNF-α and MCP-1 from alveolar epithelial cells (Figure 7D,E). When LPS-loaded alveolar cells were treated with 10–100 μg/mL YE, such induction and secretion of these inflammatory proteins were reduced (Figure 7A–E).

This study further examined whether pro-inflammatory TNF-α produced by alveolar cells might be involved in evoking alveolar emphysema by pathological stimuli. When TNF-α was treated to A549 cells, the MMP-12 protein was highly induced (Figure 7F). In contrast, ≥10 μg/mL YE blunted its induction in TNF-α-experienced alveolar cells. Thus, one can speculate that airway inflammation may be a contributor to pulmonary emphysema.

**Figure 7.** Blockade of alveolar inflammation by dried yeast extracts (YE) in lipopolysaccharide (LPS)-exposed A549 cells. Alveolar epithelial cells were incubated in media containing 2 μg/mL LPS or 10 ng/ml TNF-α in the absence and presence of 10–100 μg/mL YE for up to 24 h. Cell lysates were prepared for Western blot analysis with a primary antibody against COX-2, iNOS, ICAM-1, or MMP-12 (**A**–**C**,**F**). β-Actin protein was used as an internal control. The bar graphs (mean ± SEM, *n* = 3) represent quantitative results of the upper bands obtained from a densitometer. Alveolar secretion of TNF-α and MCP-1 was measured by using ELISA kits (**D**,**E**). Values in respective bar graphs not sharing a same small letter indicate a significant difference at *p* < 0.05.

It has been reported that sustained activation of NF-κB pathway links airway inflammation and COPD, which provides its potential as target for treatment of asthma and COPD [33]. LPS highly increased IκB phosphorylation of A549 cells, leading to induction of nuclear translocation of NF-κB (Figure 8A). The treatment of ≥10 μg/mL YE retarded its phosphorylation. Consistently, the Cy3-NF-κB staining supported the Western blot data showing nuclear translocation of NF-κB that was inhibited by YE (Figure 8B).

**Figure 8.** Involvement of NF-κB signaling in lipopolysaccharide (LPS)-induced alveolar inflammation and blockade by dried yeast extracts (YE). Alveolar epithelial cells were incubated in media containing 2 μg/mL LPS in the absence and presence of 10–100 μg/mL YE for up to 24 h. Cell lysates were prepared for Western blot analysis with a primary antibody against IκB and phospho-IκB (**A**). β-Actin protein was used as an internal control. The bar graphs (mean ± SEM, *n* = 3) represent quantitative results of the left bands obtained from a densitometer. Values in respective bar graphs not sharing a same small letter indicate a significant difference at *P* < 0.05. Immunocytofluorescence analysis was done in LPS-treated A549 alveolar epithelial cells (**B**). The NF-κB localization was identified as Cy3-red staining in cells exposed to LPS. Nuclear staining was done with DAPI (blue). Each photograph is representative of stained cells (*n* = 4). Scale bar = 50 μm.


**Author details**

Valquíria Aparecida Passos Kneipp Federal University of Rio Grande do Norte (UFRN), Natal, Rio Grande do Norte, Brazil

\*Address all correspondence to<EMAIL_ADDRESS>
© 2024 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, provided the original work is properly cited.

*New Social Practices on Brazilian TV: Reconfigurations in News Production and Distribution DOI: http://dx.doi.org/10.5772/intechopen.113307*


**2.6 Electrokinetics potential**

Passing direct current (DC) through an oil reservoir generates an electrokinetics potential which causes electrophoresis, electromigration, electrochemical reaction, electro-osmosis and Joule heating [22]. The collective actions of these mechanisms improve formation rock permeability and porosity through the dislodgement and removal of pore linings in the form of colloids, thereby increasing pore sizes and creating new flow paths [22].


**6. References**

10 Will-be-set-by-IN-TECH

Clustering groups of objects such that similar ones are placed in the same cluster, and in its application to biological datasets are very important in that it can help identification of natural groups of biological entities that might give insight about biomarkers. In this chapter, we review some clustering algorithms applied to biological data. Ensemble clustering approaches for biological data are also reviewed. Implementation of K-means, C-means and HC algorithms and merging of the algorithms using an ensemble frame work are presented using two different datasets. The datasets are protein and DLBCL-B. Two different cluster validation indices, adjusted rand and silhouette, are used for comparing the partitions from individual algorithms and ensemble clustering. Investigating Table 1, we conclude that merging individual partitions improves C-rand values meaning that ensemble approach finds partitions similar to the real partitions. Ensemble approach is coded as a Java application and

Authors thank Dilip Gautam for his contribution to this chapter. The work of ¸Sadi Evren Seker was supported by Scientific Research Projects Coordination Unit of Istanbul University, ¸

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[3] Asur, S., Ucar, D. & Parthasarathy, S. [2007]. An ensemble framework for clustering

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[6] Bandyopadhyay, S., Mukhopadhyay, A. & Maulik, U. [2007]. An improved algorithm for

protein–protein interaction networks, *Bioinformatics* 23(13): 29–40.

**5. Conclusion**

available upon request.

**Acknowledgement**

**Author details**

Sadi Evren ¸ ¸ Seker

**6. References**

*Istanbul University, Turkey*

Harun Pirim

project number YADOP-16728.

*King Fahd University of Petroleum and Minerals*

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© 2012 Wang et al., licensee InTech. This is an open access chapter 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, provided the original work is properly cited.

© 2012 Wang et al., licensee InTech. This is a paper 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, provided the original work is properly cited.

**Research on Pattern Matching with**

**Wildcards and Length Constraints:**

The practical importance of the string matching problem should be obvious to everyone. For typical word-processing applications, immense amounts of work have been done on this subject. However, with the developments in bioinformatics (Cole et al., 2005), information retrieval (Califf et al., 2003), pattern mining (Xie et al., 2010; Ji et al., 2007; He et al., 2007), etc, sequential Pattern Matching with Wildcards and Length constraints (PMWL) has attracted more and more attention. It is not difficult to think up realistic cases where PMWL plays an important role. In Dan Gusfield's book (Gusfield, 1997), they give an example about *transcription factor* to illustrate the concept of wildcard. A *transcription factor* is a protein that binds to specific locations in DNA and regulates the transcription of the DNA into RNA. In this way, production of the protein that the DNA codes for is regulated. Many transcription factors are found and can be separated into families characterized by specific substrings containing wildcards. They use *Zinc Finger*, a common transcription factor as an example. It

CYS¢¢CYS¢¢¢¢¢¢¢¢¢¢¢¢¢HIS¢¢HIS Where CYS is the amino acid cysteine and HIS is the amino acid histidine. They also give a conclusion that if the number of wildcards is bounded by a fixed constant, the problem can

Another respective example is about *promoter*. In bioinformatics, *promoter* will help researchers to quickly locate the starting position of the intron from hundreds of millions of the sequence of *ACGT*. Among these promoters, *TATA* box is a common one (Manber & Baeza-Yates, 1991). It has very loose sequence specificity, so many *TATA* sequences are not

**Methods and Completeness**

Haiping Wang, Taining Xiang and Xuegang Hu

Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/48574

has the following signature:

be solved in linear time.

**1. Introduction**


**3. Results**

After applying the inclusion and exclusion criteria, 94 women affected by vulvar EMPD and treated at the European Institute of Oncology, from October 1997 to May 2020, were selected for our retrospective analysis.

The mean age of patients at the time of first diagnosis was 63.3 years (range: 31–88) and the median follow-up time was 7 years + 10 months (range: 2 months–30 years + 7 months).

The main clinical and pathological characteristics of the enrolled women at first diagnosis and during follow-up are shown in Table 1.

Most of the patients (81%) were affected by intraepithelial EMPD (Type 1a) at first diagnosis. Invasive EMPD occurred in only 36% of cases, including 17 patients diagnosed at first occurrence and 17 during follow-up. The histology of the invasive EMPD patients is listed in Table 1.

Persistence or recurrence was very common, taking place in 86% of cases and, thus, often requiring multiple surgical instances (median: 2; range: 0–11). The histology of persistence/recurrence of EMPD is detailed in Table 1. Alternative treatments were applied, especially in the case of relapse, including local therapy with imiquimod (63%), photodynamic therapy (5%) and radiotherapy (12%). *Diagnostics* **2023**, *13*, 464 5 of 12

> After excluding one patient with an unknown death date, the 5 year overall survival was 90.5% (95% CI: 81.8–95.1%), as shown in Figure 1.

(95% CI: 4.8–27.8%) at 15 years, respectively (Figure 2).

sidering death as a competing event (N = 93).

**Figure 1.** Overall survival of women affected by vulvar EMPD (N = 93). **Figure 1.** Overall survival of women affected by vulvar EMPD (N = 93).

Overall, nine women developed CV localization of EMPD, with a cumulative inci‐ dence of 2.5% (95% CI: 0.5–8.0%) at 5 years, 6.5% (95% CI: 1.9–15.1%) at 10 years and 14.0%

**Figure 2.** Cumulative incidence function of cervico‐vaginal (CV) localization of vulvar EMPD, con‐

**Table 1.** Clinical and pathological characteristics of women affected by vulvar EMPD at first diagnosis and during follow-up (N = 94).

*Diagnostics* **2023**, *13*, 464 5 of 12

Overall, nine women developed CV localization of EMPD, with a cumulative incidence of 2.5% (95% CI: 0.5–8.0%) at 5 years, 6.5% (95% CI: 1.9–15.1%) at 10 years and 14.0% (95% CI: 4.8–27.8%) at 15 years, respectively (Figure 2). dence of 2.5% (95% CI: 0.5–8.0%) at 5 years, 6.5% (95% CI: 1.9–15.1%) at 10 years and 14.0% (95% CI: 4.8–27.8%) at 15 years, respectively (Figure 2).

**Figure 2.** Cumulative incidence function of cervico‐vaginal (CV) localization of vulvar EMPD, con‐ sidering death as a competing event (N = 93). **Figure 2.** Cumulative incidence function of cervico-vaginal (CV) localization of vulvar EMPD, considering death as a competing event (N = 93).

The main characteristics of women who developed CV involvement from vulvar EMPD are reported in Table 2, including histology at diagnosis and at vulvar recurrence and the timing and type of CV localization. The majority of women (6/9) showed an intraepithelial vulvar EMPD (Type 1a) at first diagnosis, but three of them developed an invasive disease (Type 1b) at recurrence. The CV localizations occurred after a median time of 133 months (range: 16–334) from the first diagnosis of vulvar EMPD. All cases except one were firstly detected by abnormal pap smear and histologically confirmed by cervical and vaginal colposcopic-guided biopsies.

Most patients (5/9) developed invasive EMPD of the cervix and/or vagina. In the case of invasive or unusual disease, magnetic resonance imaging (MRI) of the lower abdomen and total body positron emission tomography (PET) were performed to rule out pelvic node or distant metastases. Interestingly, one woman with vulvar EMPD Type 1a at the onset developed invasive cervico-vaginal involvement with node metastasis after 251 months from the initial diagnosis. CV intraepithelial EMPD occurred in only two patients, among which one later developed invasive EMPD of the urethra. Two CV localizations manifested with other associated diseases: invasive urothelial carcinoma and invasive mucinous intestinal-type adenocarcinoma with node metastasis.

Most of the women underwent hysterectomy with partial or total colpectomy based on the site of extravulvar EMPD localization. The patient with node metastases was treated with chemotherapy in association with anti-HER2 (human epidermal growth factor 2) targeted monoclonal antibodies. Instead, the patient diagnosed with cervical invasive mucinous intestinal-type adenocarcinoma refused any treatment because of comorbidities and died nine months after the diagnosis. Radiotherapy was offered to the woman who developed an unresectable and advanced form of invasive EMPD of the urethra.


*3.2.1 Gene silencing*

Similar to miRNAs, tRFs have been associated with the epigenetic regulator RISC, however mechanistic details on the role of tsRNA in the RISC remain to be elucidated. A recent meta-analysis of short RNA libraries from HEK293 cells demonstrated that both tRF-3 and tRF-5 associate with Ago proteins; however, a preference for Ago1, 3 and 4 over Ago2 was identified [59]. Interestingly a subsequent study in *D. melanogaster* revealed an age-related shift in tRF-Ago binding demonstrating a preference for Ago2 binding over Ago1 with increasing age [60].

### *3.2.2 Regulation of protein translation*

The synthesis of protein is a central activity in all cells that consumes a high level of energy and is dynamic in response to metabolic conditions and external stimuli. The regulation of protein translation therefore is a vital process in the maintenance of cell viability and the stress response. Canonical cap-dependent translation begins with the formation of the eukaryotic initiation factor (eIF) 4F complex containing eIF4A, a DEAD-box helicase, eIF4E and eIF4G. The eIF4E subunit binds to the 5' m7GTP cap on target mRNA and the eIF4G subunit is a scaffold protein that mediates the recruitment of other proteins including eIF3 and poly(A) binding protein (PABP). eIF4F binding to the 5′ m7GTP cap and the 3′ poly(A) tail circularizes the target mRNA and allows the 48S pre-initiation complex, containing the 40S small ribosomal subunit, Met-tRNAi met and eIF2, to scan the 5′ untranslated region and find the AUG start codon [61].

The dynamic regulation of protein translation in response to cellular stress and metabolic conditions is vital to cell survival. Stress-induced Ang-generated 5′tiRNA have been shown to halt the initiation of protein translation and facilitate the packaging of stalled translational complexes into stress granules [54]. Stress granules are cytoplasmic RNA-protein complexes that rapidly assemble and disassemble in response to cellular stress. This sequestration allows for the utilization of energy stores elsewhere and the recommencement of protein translation under optimum conditions [62]. Specific 5′tiRNA that contain a terminal oligoguanine (TOG) motif form stable G-quadruplex (G4) structures that directly bind the HEAT domain of eIF4G displacing eIF4A and inhibiting scanning of the mRNA target. Furthermore, 5′tiRNA with a 5′ monophosphate modification have been shown to bind the RNA binding protein YB-1 via the cold shock domain to precipitate the formation of stress granules [63].

Current knowledge on the effect of tRFs on protein translation is less advanced. Research in prokaryotic cells has demonstrated that tRF-5c of Val-GAC can bind the small ribosomal subunit and interfere with peptidyl transferase activity thereby inhibiting protein translation [64, 65]. In eukaryotic cells the tRF-3b of Gly-GCC reduced the level of specific protein with no concomitant reduction on mRNA levels indicating regulation at the translational level [66].

#### **3.3 tsRNA functions**

#### *3.3.1 tsRNA and apoptosis*

Disruption to the tsRNA system has been associated with increased cell death. Hypo-methylation of tRNA that arises from the inhibition of NSun2 increases cleavage by Ang and the accumulation of 5′tiRNA. The subsequent sustained depression in protein translation results in neuronal shrinkage, impaired synapse formation, cell death and is associated with neurodevelopmental deficiencies [67]. Loss-of-function mutations in the RNA kinase CLP1 has been shown to increase the level of tyrosine pre-tRNA fragments resulting in exaggerated p53 activation and vulnerability to cell death in cells exposed to oxidative stress [68]. Conversely, Ang has been shown to reduce cell death in neurons exposed to hyperosmotic stress in a tiRNA-mediated fashion. Specific Ang-generated tiRNA interact with cytochrome c and form a ribonucleoprotein complex that limits the formation of apoptosomes and reduces caspase-3 activation [53].


**3. The SFE2D architecture**

To separate the spectral features, we created a 2D feature extraction scheme that combines 2D CWT with variance, kurtosis, and negentropy maximization algorithms (PCA, kICA, and nICA) to extract spectral features from wavelet spectra. The algorithm consists of three main stages (**Figure 5**):

An effective spectral feature extraction depends on computer hardware specifications, specifically for larger numbers of spectral features produced by increasing the number of scales and mother wavelet's directions. In practice, users can create a larger number of features by changing the CWT parameters in the SFE2D program.

**Figure 5.** *Schematic view of the SFE2D procedure.*

For large numbers of CWT features (100–1000 CWT features), PCA/ICA methods provide a way to decorrelate and separate spectral features from CWT images.

If needed, users can also reduce the dimensionality to summarize the features. By running the proposed workflow in the SFE2D program, the redundant frequency volumes could be reduced to a more manageable number of components. Taking advantage of the ICA statistical properties, we can keep the most geologically pertinent information within the spectral decomposed data.


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Pharmacokinetics and Pharmacodynamics for Treatment Optimization in Clinical

### *Edited by Ayman Noreddin*

This book, "Readings in Advanced Pharmacokinetics - Theory, Methods and Applications", covers up to date information and practical topics related to the study of drug pharmacokinetics in humans and in animals. The book is designed to offer scientists, clinicians and researchers a choice to logically build their knowledge in pharmacokinetics from basic concepts to advanced applications. This book is organized into two sections. The first section discusses advanced theories that include a wide range of topics; from bioequivalence studies, pharmacogenomics in relation to pharmacokinetics, computer based simulation concepts to drug interactions of herbal medicines and veterinary pharmacokinetics. The second section advances theory to practice offering several examples of methods and applications in advanced pharmacokinetics.

Readings in Advanced Pharmacokinetics - Theory, Methods and Applications

Readings in Advanced

Pharmacokinetics

Theory, Methods and Applications

*Edited by Ayman Noreddin*

Photo by Totojang / iStock


**4. Conclusions**

Within the ESA business applications demonstration project SnowSense (2015–2018), we successfully demonstrated a large scale snow hydrological monitoring service, by combining newly developed in situ stations based on signals of the Global Navigation Satellite System (GNSS), Earth Observation (EO) and hydrological modelling. With this combined approach, we present a reliable, and cost-efficient tool for the determination of snow cover properties like snow water equivalent (SWE), snow liquid water content (LWC), snow extent as well as run-off assessment, for real-time and forecast applications.

The GNSS in situ component was successfully applied and validated at the well-equipped study site NEIGE at Forêt Montmorency, Quebec, Canada. Furthermore, the entire SnowSense service providing modelled, in situ-, and EO-assimilated run-off was applied and validated at four run-off gauges within the Humber River and the Exploit River catchments on the island of Newfoundland, Canada.

The entire SnowSense service solution driven with an integrated numerical weather prediction (NWP) model for its application in this study in Newfoundland is capable of providing detailed knowledge on water stored as snow over large spatial scales. It is able to provide real-time and forecasted snow and run-off information and, if desired, also on reservoir status, which might be of great interest for hydropower plant operators. This information, which can be provided in various time steps, e.g., hourly up to daily, is especially needed in regions or catchments where in situ stations are only sparsely or non equipped catchments. The service is applicable at almost any location and was especially designed for remote locations, where access is limited and snow and run-off measurements were difficult up to now.

Within the project, the SnowSense service already reached a market dedicated design, based on the identification of potential customers (i.e., hydropower plants) and use cases (i.e., weather and climate observations, e.g., by national weather services).

**Author Contributions:** F.A., H.B. and W.M. conceived and developed the project idea; F.K., M.L., P.H. and F.A. were deeply involved in the development of the SnowSense algorithm, hardware development, and model integration; P.K. and H.B. were responsible for the PROMET model setup and the EO data assimilation. F.A. was the SnowSense project manager; F.A., P.H., F.K. and W.M. are involved in the patenting of the algorithm and the hardware; A.R., F.A. and F.K. wrote the paper.

**Funding:** The project was co-funded by the European Space Agency (ESA, 000113149/14/NL/AD) within the Business Application Demonstration Project SnowSense (2015–2018) (https://artesapps.esa.int/projects/ snowsense-dp).

**Acknowledgments:** We thank the editors for their work and the two anonymous reviewers for their constructive comments. Florian Appel, Franziska Koch, Patrick Henkel, Markus Lamm, Philipp Klug and Anja Rösel were co-funded by the European Space Agency (ESA, 000113149/14/NL/AD) within the Business Application Demonstration Project SnowSense (2015–2018) (https://artesapps.esa.int/projects/snowsense-dp), which is gratefully acknowledged. The authors would like to thank all involved staff members of the demo users Nalcor/NL Hydro and the Department of Municipal Affairs and Environment, Water Resources Management Division (WRMD) of the Government of Newfoundland and Labrador, for their active support to the project, and for providing their run-off data. We thank the staff from University Laval for the opportunity to set up our sensor at the NEIGE site in Forêt Montmorency and for the opportunity to participate in the inter-comparison study. For the developed algorithm for the GNSS-based SWE and LWC determination as well as the invention of the in situ hardware, two patents are pending. SnowSense -R is a registered trademark.

**Conflicts of Interest:** The authors declare no conflict of interest.


**Chapter 5**

*Natural Hazards - Impacts, Adjustments and Resilience*

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**68**