*Article* **Improving the Safety of DIEP Flap Transplantation: Detailed Perforator Anatomy Study Using Preoperative CTA †**

**Katharina Frank 1 , Armin Ströbel 2 , Ingo Ludolph 1 , Theresa Hauck 1 , Matthias S. May 3 , Justus P. Beier 1,4 , Raymund E. Horch 1 and Andreas Arkudas 1,\***

**Abstract:** Background: Deep inferior epigastric perforator and muscle sparing transverse rectus abdominis muscle flaps are commonly used flaps for autologous breast reconstruction. CT-angiography allows to analyse the perforator course preoperatively. Our aim was to compare the different aspects of perforator anatomy in the most detailed study. Methods: CT-angiographies of 300 female patients with autologous breast reconstruction of 10 years were analysed regarding the anatomy of the deep inferior epigastric artery and every perforator. Results: Overall, 2260 perforators were included. We identified correlations regarding the DIEA branching point and number of perforators and their intramuscular course. The largest perforator emerged more often from the medial branch of the DIEA than the smaller perforators (70% (416/595) vs. 54% (878/1634), *p* < 0.001) and more often had a direct connection to the SIEV (large 67% (401/595) vs. small 39% (634/1634), *p* < 0.01). Medial row perforators were larger than the laterals (lateral 1.44 mm ± 0.43 (*n* = 941) vs. medial 1.58 mm ± 0.52 (*n* = 1304) (*p* < 0.001)). The larger and more medial the perforator, the more likely it was connected to the SIEV: perforators with direct connection to the SIEV had a diameter of 1.65 mm ± 0.53 (*n* = 1050), perforators with indirect connection had a diameter of 1.43 ± 0.43 (*n* = 1028), perforators without connection had a diameter of 1.31 mm ± 0.37 (*n* = 169) (*p* < 0.001). Medial perforators were more often directly connected to the SIEV than lateral perforators (medial 56% (723/1302) vs. lateral 35% (327/941), *p* < 0.001). A lateral perforator more often had a short intramuscular course than medial perforators (69% (554/800) vs. 45% (474/1055), *p* < 0.001), which was also more often observed in the case of a small perforator and a caudal exit of the rectus sheath. Conclusion: The largest perforator emerges more often from the medial branch of the DIEA and frequently has a direct connection to the SIEV, making medial row perforators ideal for DIEP flap transplantation.

**Keywords:** CTA; autologous breast reconstruction; DIEP flap; MS-TRAM flap; perforator


**2.2 Ceramic battery technology**

They are all solid-state batteries made up of ceramic materials as their electrolytes. The demand for high-density batteries has necessitated research into more technologies that will enhance the thermal stability, mechanical strength and wettability of solid-state battery manufacture. Enhancements at the metal electrodes and/or separator boundaries are made possible by the introduction of ceramic batteries to boast some of the desired properties needed for solid-state batteries. Ceramic-based flexible sheet electrolytes have been formed to improve the energy density of solid-state batteries by synthesizing flexible composite Al-doped LLZO sheet electrolyte [17], coating of Al2O3, SiO2 and TiO2 onto polyethylene membrane seperators [18]. They

**Figure 2.**

*Classification of printed battery technology.*

are used in cogeneration and fuel cell, electric vehicles, mobile devices and stationary storage applications [19].

#### **2.3 Lithium polymer battery technology**

This works on the principle of intercalation and de-intercalation of lithium ions from the positive electrode (cathode) to the negative electrode (anode) and vice versa through a solid-state electrolyte that provides the conductive medium. Lithiumpolymer batteries are lightweight, long-lasting and powerful solid-state batteries that can guarantee a constant energy supply [20]. They have a high energy density, flat voltage curves, low self-discharge and no memory effect from being discharged and charged again. The technology is used in laptop computers, personal electronics, cellular phones [21], notebooks and digital cameras. The polymer electrolyte may be in form of dry solid polymer electrolyte, polymer in-salt system, single-lithium-ion conducting electrolyte and gel polymer electrolyte [22].

#### **2.4 Nickel-metal hydride (NiMH) button battery technology**

This technology is an energy storage system that depends on charge/discharge reactions occurring between the nickel oxide-hydroxide cathode as the active material and the hydrogen-absorbing alloy anode [23]. This battery technology exhibits good tolerance to overcharge/discharge, high power capacity, very safe and compatible [24]. They are mainly used as power sources for hybrid electric vehicles, digital cameras and cell phones [25].

### **3. Deposition techniques**

Thin-film deposition techniques are used to modify the surface properties of solidstate thin-film batteries. The modification affects the battery characteristics such as energy density, conductivity, storage capacity, charging and discharge time, etc.

Thin-film battery deposition techniques are classified into physical and chemical deposition processes as shown in **Figure 3** [26]. In the chemical deposition process, a chemical reaction takes place before the product is deposited on the substrate while the physical deposition method involves only physical mixing and deposition of the mixture on the substrate without any chemical reaction [27]. The coating material in a physical deposition is always a solid while the coating materials in chemical deposition are always in gaseous form. Physical deposition techniques include thermal evaporation, sputtering, ion plating and arc vapor deposition while chemical deposition techniques include chemical vapor deposition, plating, sol-gel deposition, chemical bath deposition and spray pyrolysis deposition techniques [28].

#### **3.1 Physical deposition method**

This method includes thermal evaporation, molecular beam epitaxy, pulsed laser deposition, ion plating evaporation, cathodic arc deposition and sputtering techniques. In thermal evaporation, molten metals or metal oxides form vapor and are deposited on the substrate as it cools [29, 30]. Molecular beam epitaxy (MBE) is a deposition technique in which a single crystal layer is deposited in a single crystal substrate, using molecular beams in an ultra-high vacuum chamber [31]. It is used in *Thin-Film Batteries: Fundamental and Applications DOI: http://dx.doi.org/10.5772/intechopen.109734*

**Figure 3.** *Deposition techniques.*

multi-junction solar cell applications [32], Ga-FACE GaN electron devices [33], etc. Pulsed laser deposition (PLD) process is a technique where a high-power pulsed laser beam strikes a target, vaporizes it and deposits it as a thin film on a substrate [34]. It is useful for optical applications, especially for UV laser emission [35] and superconductor devices for electronic and medical applications [36].

#### **3.2 Chemical deposition method**

Chemical deposition of thin films involves the generation of the atoms, molecules or ions through a chemical means; transportation of the atoms or ions through a medium and condensation and cooling of the atoms, molecules or ions on the substrate [27]. Chemical methods include chemical vapor deposition (CVD), electrodeposition, electroless deposition, spray pyrolysis, ionic layer deposition, sol-gel technique, chemical bath, spray and spin coating as shown in **Figure 2**.

Spray pyrolysis is the process of depositing a thin film on a heated surface by spraying a solution on the heated substrate [37]. The solution reacts with the heated substrate to form a thin film on the surface of the substrate. Mostly used for manufacturing semi-conductor alloys and conductive glasses. Electrodeposition is a technique of coating a thin layer of metal on top of a different metal through electrolysis by reducing the cations of the material from the electrolyte and depositing it as a thin film on the substrate [38]. It is a major method for the production of rechargeable lithium-ion batteries [39, 40]. Electroless deposition is a thin-film coating technique without the application of external electric power. It is used for non-conducting substrates and its major application is in the metalizing of printed wiring boards [41]. Atomic layer deposition (ALD) is a chemical deposition technique where chemical precursors introduced on the surface of the substrate react to form ultra-thin film monolayers on the substrate surface. They find application in fuel cells, capacitors,

microelectronics and areas where highly uniform ultra-fine mono-layered thin films are required [42]. Sol-gel is a technique where metal alkoxides, used as precursors, are dissolved in a solvent (especially water or alcohol), heated and stirred to form a gel, which is condensed, dried and deposited on the substrate as a thin film [43]. The process involves hydrolysis, polycondensation, gelation, drying and crystallization [44].

#### **3.3 Deposition technique for cathode materials in thin-film batteries**

A cathode is a very important component of thin-film batteries. This electrode material facilitates the stability of the electrochemical reactions in the electrode/electrolyte interface. The material composition of the cathode determines the thermal stability and rate capacity of the battery. For a good and efficient battery system, there is always a good ion transport mechanism between the anode and the cathode through the electrolyte and the acquisition and flow of electrons through the external circuit during an electrochemical reaction. Different deposition methods have been used in the design and fabrication of cathode materials for different solid-state battery applications. Production of LiCoO2 cathode materials using radio frequency (RF) magnetron sputtering deposition technique for micro battery applications was studied by Jullien et al. [45], MoO3 thin-film cathodes using DC sputter deposition technique for Li-ion rechargeable battery applications [39] and V2O5 thin-film cathodes using radio frequency (RF) reactive sputtering technique for high-performance thin-film batteries (TFB) [46].

#### **3.4 Deposition technique for anode materials in thin-film batteries**

A good anode material should be a good conductor and reducing agent and possess a high electrical energy density. They are made from carbon (graphite), lithium metals, lithium alloys, titanium and its alloys, silicon and other metals. Several methods have been developed on good anode materials for solid-state thinfilm batteries for micro-mechanical system applications. Reto Pfenninger et al. [47] studied the electrodeposition of Li-garments on solid state anode. The group worked on metal oxide anodes of Li4Ti5O12 with good ionic conductivity and less Li-dendrite formation for microbattery applications [47]. Pulsed electrodeposition of Sn-Cu anodes has been studied and developed for enhanced cycle performance in micro battery applications [48, 49]. Spray deposition of silicon anode materials with high energy density for solid-state battery application was developed by Shin Kimura et al. [50]. Many researchers have fabricated different anode materials and their reports have shown that anodes fabricated with good properties have shown to exhibit better battery performances. Li4Ti5O12 anode thin film was deposited on a magnesium oxide (MgO) substrate using the pulse laser deposition (PLD) technique [47], pulsed laser deposition method was used to fabricate a silicon thin-film anode with iron sulphide for solid-state battery applications [51]. DC magnetron sputtering technique was reported by L. Baggetto et al. for Cu2Sb thin-film deposition for sodium ion (Na-ion) batteries because of excellent storage capacity, high-rate capacity, good reaction potential and decent cycling capacity retention [52]. Silicon/carbon (SiC) thin-film anode was fabricated using a radio frequency (RF) magnetic sputtering technique with improved cycling performance and high current density for thin-film battery applications.


*Edited by Alexander Vakhrushev*

This book is devoted to the description of research and design of photonic crystals. Topics included in the book cover a wide range of research in the field of theoretical analysis and experimental investigation: the electromagnetic field in the photonic crystal, propagation of waves in the gyrotropic magnetophotonic crystals, low onephoton absorption, ultratransparent photonic crystals, colloidal assembly, photonic crystal application for development of all-optical computational system, design strategies for PC devices, self-organization of liquid crystalline nanostructures, and optical diodes. This book will be useful for engineers, technologists, researchers, and postgraduate students interested in the research, design, fabrication processes, and applications of photonic crystals.

ISBN 978-953-51-3961-4

Theoretical Foundations and Application of Photonic Crystals

Photo by rep0rter / iStock


**3. Results**

#### *3.1. Experimental Configuration and Evaluation Metrics*

All experiments were conducted on a server running the Ubuntu 16.04 operation system. The server was equipped with two Tesla p40 GPUs and a Xeon Gold 5118 CPU. The resolutions of images from the dataset were resized to 560 × 420. Due to the utilization of transfer learning, the model converged rapidly, and the number of training epochs was set to 10 while each branch was separately trained. When two branches were trained jointly, the number was increased to 20. For all experiments, the initial learning rate was set to 0.0001 and the Adam optimizer was used. Additionally, 5-fold cross validation was implemented. The training group with fold *K* set for testing was named training group *K*.

Intersection over union (IoU) was utilized as the metric form of segmentation tasks of this paper to evaluate the accuracy of the outputs. IoU is calculated as follows:

$$IoI = \frac{TP}{TP + FP + FN} \tag{5}$$

In experiments of the foreground extraction task, only the IoU of foreground that represented bodies of target vehicles were counted.

In the EV fire trace recognition task, the number of classes was set to 4, so the mean IoU (mIoU) of 4 classes was calculated to evaluate the performance. As discussed in 3.1, the 4 classes were IN, MB, SB, BG, and the mIoU could be calculated as follows:

$$mIoI = \frac{1}{4}(IoI\_{BG} + IoI\_{IN} + IoI\_{MB} + IoI\_{SB})\tag{6}$$

Additionally, to evaluate the accuracy of vehicle body segmentation, the union regions of IN, MB and SB were regarded as "Vehicle Body" (VB) regions; to evaluate the segmentation accuracy of burnt regions as a whole, the union of MB regions and SB regions were regarded as "Fire Trace" (FT) regions. Their IoU was thus calculated as follows:

$$IoL\_{VB} = \frac{I\_{IN \cup MB \cup SB}}{I\_{IN \cup MB \cup SB}} \tag{7}$$

$$IoU\_{FT} = \frac{I\_{MB\cup SB}}{I\_{MB\cup SB}}\tag{8}$$


**A Critical Risk Factor for a Major Side Effect of Interferon-Alpha Therapy: Activated Indoleamine 2,3- Dioxygenase 1 is Related to Depressive Symptoms Interferon-Alpha Therapy: Activated Indoleamine 2,3-Dioxygenase 1 is Related to Depressive Symptoms**

**A Critical Risk Factor for a Major Side Effect of**

DOI: 10.5772/intechopen.71013

Yuki Murakami and Yukio Imamura Additional information is available at the end of the chapter

Yuki Murakami and Yukio Imamura

Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/intechopen.71013

### **Abstract**

[79] Ishii T, Ichikawa T, Minoda K, Kusaka K, Ito S, Suzuki Y, Akagawa M, Mochizuki K, Goda T, Nakayama T. Human serum albumin as an antioxidant in the oxidation of ()-epigallocatechin gallate: Participation of reversible covalent binding for interaction and stabilization. Bioscience, Biotechnology, and Biochemistry. 2011;75:100-106. DOI: 10.1271/

[80] Ishii T, Minoda K, Bae MJ, Mori T, Uekusa Y, Ichikawa T, Aihara Y, Furuta T, Wakimoto T, Kan T, Nakayama T. Binding affinity of tea catechins for HSA: Characterization by highperformance affinity chromatography with immobilized albumin column. Molecular

[81] Zhu M, Chen Y, Li RC. Oral absorption and bioavailability of tea catechins. Planta Medica.

[82] Kohri T et al. Synthesis of ()-[4-3H]epigallocatechin gallate and its metabolic fate in rats after intravenous administration. Journal of Agricultural and Food Chemistry. 2001;49:

Nutrition & Food Research. 2010;54:816-822. DOI: 10.1002/mnfr.200900071

bbb.100600

2000;66:444. DOI: 10.1055/s-2000-8599

36 Pharmacokinetics and Adverse Effects of Drugs - Mechanisms and Risks Factors

1042. DOI: 10.1021/jf0011236

Hepatitis C virus (HCV) infection affects approximately 170 million people worldwide. Interferon-alpha (IFN-α) is a cytokine that is related to early viral infection and has both antiviral and antiproliferative properties. The current standard treatment for long-term chronic hepatitis C (CHC) consists of combination therapy with IFN-α and ribavirin, which has a broad spectrum antiviral effect. Despite the potential therapeutic benefits of IFN-α, its administration often causes many side effects, such as somatic and neuropsychiatric symptoms. Depression is a serious and frequently occurring side effect of IFN-α therapy, and this is one of the major reasons for cessation of the therapy. Therefore, in order to avoid the discontinuation of INF-α therapy owing to depressive symptoms, it is important to identify the risk factor(s) leading to the onset of associated depressive symptoms. In this chapter, we introduce our novel findings on the association between IFN treatment and the onset of depression in CHC patients as well as the potential neurobiological mechanisms by which depression may arise. We also highlight a potential approach for predicting the onset risk of depression as a side effect in these patients.

**Keywords:** hepatitis C, IFNs, depression, tryptophan catabolism, indoleamine 2,3-dioxygenase 1


*3.1.1 Visual inspection*

It is an intraoral examination under light with the help of a mirror. In addition to being a frequently used method, it is insufficient to diagnose caries formations at the initial stage [14].


**5. Conclusions**

The prenatal diagnosis of CHD in the last 30 years has reached a high degree of diagnostic accuracy allowing identification of almost all the main forms of CHD

during fetal life, in expert centers. In particular, prenatal diagnosis of conditions that constitute a neonatal emergency, such as duct dependent lesions, allows to manage the delivery in a tertiary care center. This strategy improves survival significantly, and reduces preoperative morbidity, and the risk of neurological compromise. While some CHD may have a successful surgical correction in postnatal life, a small selective subset of these defects can progress during intrauterine life and be susceptible to early interventions in the uterus. Fetal cardiac interventions might prevent in utero worsening from a simple, potentially correctable lesion to a complex cardiac condition in selective patients. FCIs are most commonly and successfully performed in the setting of 1) fetal aortic stenosis with risk for evolving to hypoplastic left heart syndrome, 2) HLHS with intact or restrictive atrial septum 3) pulmonary atresia/ stenosis with intact ventricular septum, with specific features associated with risk for worsening right ventricular hypoplasia. FCI also includes transplacental therapy, including maternal antiarrhythmic drugs in case of fetal arrhythmias and steroids. FCI is a procedure that has maternal and/or fetal risks, which must be well explained to parents at the time of counseling, to be sure that they fully understand the possible complications.

Moreover, fetal early diagnosis allows to program earlier clinical management and surgery after birth, improving outcomes. The improvement in the diagnosis and treatment of these conditions, due to extraordinary advances in imaging and cardiac surgical and interventional technologies in the last twenty years, has led to a significant increase in survival rate in patients with CHD, and accordingly, the number of patients with CHD who have reached adulthood has increased significantly and even surpass the number of affected kids.


**Treatment of Influenza**

[55] Ivashkiv LB, Donlin LT. Regulation of type I interferon responses. Nature Reviews.

[56] Groettrup M, Kirk CJ, Basler M. Proteasomes in immune cells: More than peptide pro-

[57] Stanley ER, Guilbert LJ, Tushinski RJ, Bartelmez SH. CSF-1 -a mononuclear phagocyte lineage-specific hemopoietic growth factor. Journal of Cellular Biochemistry. 1983;**21**:

[58] Tomljenovic L, Shaw CA. Mechanisms of aluminum adjuvant toxicity and autoimmu-

[59] Trinchieri G. Type I interferon: Friend or foe? The Journal of Experimental Medicine.

[60] Sasaki E, Momose H, Hiradate Y, Furuhata K, Takai M, Asanuma H, et al. Modeling for influenza vaccines and adjuvants profile for safety prediction system using gene expres-

[61] Szymański P, Markowicz M, Mikiciuk-Olasik E. Adaptation of high-throughput screening in drug discovery-toxicological screening tests. International Journal of Molecular

Immunology. 2014;**14**:36-49

130 Influenza - Therapeutics and Challenges

151-159

2010;**207**:2053-2063

Sciences. 2012;**13**:427-452

ducers? Nature Reviews. Immunology. 2010;**10**:73-78

nity in pediatric populations. Lupus. 2012;**21**:223-230

sion profiling and statistical tools. PLoS One. 2018;**13**:e0191896

**Chapter 7**

Provisional chapter

**Therapeutic Approach for Seasonal Influenza and**

DOI: 10.5772/intechopen.76473

Influenza infection is usually a self-limiting and suddenly life-threatening disease. Seasonal influenza causes severe clinical symptoms and almost subsides within 7 days in patients without severe illness, following no complications of pneumonia and encephalitis. Influenza A (H1N1) pdm09 brought the disaster including many deaths. We cannot make differential diagnosis between seasonal and pandemic influenza adequately in a pre-pandemic state. Seasonal influenza displaces suddenly pandemic, and we necessarily establish a standard treatment for influenza viral infection in routine work. If antiviral therapy would not be effective for patients with influenza viruses in an early period of illness, further investigations would be proceeded concerning three points: mutations of influenza viruses resistant to neuraminidase inhibitors (NAIs), concomitant diseases of patients, and a new pandemic virus. If the systemic procedure would be functioned, we are able to reduce individually burden of patients with severe clinical symptoms and leading complications and socially delay widespread of pandemic and plan for the

Keywords: antiviral therapy, seasonal epidemic, pandemic, streamline surveillance,

Influenza viruses spread seasonally and cause infection of the airway tract in humans following severe symptoms. Influenza viruses grow and multiply among in human, swine, and avian bodies. Influenza viruses escape human immunological protective system against influenza viruses by changing their epitope detected by human immune cells. Annual seasonal influenza epidemic often happened under no antiviral procedure by easy infection of influenza viruses.

> © 2016 The Author(s). Licensee InTech. 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 eproduction in any medium, provided the original work is properly cited.

© 2018 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.

Therapeutic Approach for Seasonal Influenza and

Additional information is available at the end of the chapter

streamline management of pandemic documents.

the systemic procedure

1. Introduction

Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/intechopen.76473

**Pandemic**

Pandemic

Yuji Takemoto

Abstract

Yuji Takemoto

#### **Therapeutic Approach for Seasonal Influenza and Pandemic** Therapeutic Approach for Seasonal Influenza and Pandemic

DOI: 10.5772/intechopen.76473

#### Yuji Takemoto Yuji Takemoto

Additional information is available at the end of the chapter Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/intechopen.76473

#### Abstract

Influenza infection is usually a self-limiting and suddenly life-threatening disease. Seasonal influenza causes severe clinical symptoms and almost subsides within 7 days in patients without severe illness, following no complications of pneumonia and encephalitis. Influenza A (H1N1) pdm09 brought the disaster including many deaths. We cannot make differential diagnosis between seasonal and pandemic influenza adequately in a pre-pandemic state. Seasonal influenza displaces suddenly pandemic, and we necessarily establish a standard treatment for influenza viral infection in routine work. If antiviral therapy would not be effective for patients with influenza viruses in an early period of illness, further investigations would be proceeded concerning three points: mutations of influenza viruses resistant to neuraminidase inhibitors (NAIs), concomitant diseases of patients, and a new pandemic virus. If the systemic procedure would be functioned, we are able to reduce individually burden of patients with severe clinical symptoms and leading complications and socially delay widespread of pandemic and plan for the streamline management of pandemic documents.

Keywords: antiviral therapy, seasonal epidemic, pandemic, streamline surveillance, the systemic procedure

#### 1. Introduction

Influenza viruses spread seasonally and cause infection of the airway tract in humans following severe symptoms. Influenza viruses grow and multiply among in human, swine, and avian bodies. Influenza viruses escape human immunological protective system against influenza viruses by changing their epitope detected by human immune cells. Annual seasonal influenza epidemic often happened under no antiviral procedure by easy infection of influenza viruses.

© 2016 The Author(s). Licensee InTech. 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 eproduction in any medium, provided the original work is properly cited. © 2018 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.

Seasonal influenza viruses affect 10–20% of human population in epidemics each year [1] and worldwide, cause an estimated 3.5 million cases of severe illness and 250,000–500,000 death each year [2]. Although almost infected patients with seasonal influenza viruses recover from the disease in less than 2 weeks. On the other hand, some group of people containing young children, adults being elder than 65 years old and compromised hosts with severe illness had complications of influenza infection leading hospital admission [3]. Influenza viruses mutate and change the disease severity in host when transferring from avian to swine or from avian to human or from swine to human. A new mutant of influenza viruses is defined at the site of a mutation and called as influenza A (H5N1), A (H7N9), and A (H1N1) pdm09 each, causing severe affected people following many deaths [4]. Three influenza pandemics happened in the twentieth century: in 1918–1919, 1957, and 1968 and were called as Spanish flu, Asian flu, and Hong-Kong flu each and caused the severe disaster [4]. Especially Spanish flu brought estimated 20–50 million mortality [4]. We have the inability to predict and testify the appearance of dangerous influenza viruses to human health by the lack of rapid, affordable, highly sensitive, and specific diagnostic tests. The appearance of a new mutation of influenza viruses was noticed as unsuccessful treatment cases leading to life-threatening complications of influenza infection in the treatment of seasonal influenza [4]. The expansion of disaster by both the delayed use and little sharing of pre-pandemic information makes difficult in minimizing a widespread of a new mutant infection of influenza virus [4]. So it is necessary to establish a systemic procedure of diagnosis and treatment for patients with seasonal influenza and pandemic viruses in early phase of pandemic. In a first step of diagnosis of influenza virus infection, we can use rapid diagnostic kits for influenza A/B virus and we diagnose easily seasonal influenza infection in the outpatients. But rapid immune-chromatographic kits cannot show the mutation of influenza virus subtypes, we cannot make a differential diagnosis between seasonal and pandemic influenza virus by it [5]. A mutation of influenza virus subtypes is evaluated by reverse-transcriptase polymerase chain reaction (RT-PCR) and direct sequence of a recognition site of influenza virus subtypes [5]. This method is expensive and time-consuming. We cannot apply this method to all outpatients with influenza virus. On the course of clinical treatment, we need to discriminate patients with suspicious pandemic influenza virus from the other patients with seasonal influenza effectively and economically. We would discuss the feasibility and execution of this trial in the following chapters.

influenza vaccine. Recently, WHO has been recommending QIV in 2013 and QIV has been adopted in several countries. QIV is estimated cost-effective and cost saving to reduce the burden of outpatient visit for influenza but 1.62 hospitalizations and 0.078 deaths per 100,000 individuals were estimated in Japan [9]. VE is often influenced by the timing between vaccination and influenza epidemic. Early epidemic occurs seasonally before administration of vaccine against seasonal influenza infection in people and VE is low. A mismatch of influenza viral strains between vaccination and epidemic makes VE low. It is very difficult to overcome influenza infection only by vaccination because of current VE and variability of influenza virus. We need the adequate diagnosis and treatment systematically after the procedure of

Therapeutic Approach for Seasonal Influenza and Pandemic

http://dx.doi.org/10.5772/intechopen.76473

135

Adoption of neuraminidase inhibitors (NAIs) for treating patients with influenza virus improves clinical incidences of outpatients leading hospitalization. Influenza infection is typical symptoms such as fever, headache, sore throat, cough, joints lasting, and sometimes diarrhea and nausea. On the first step of the treatment for influenza, concomitant use of antipyretic and mild analgesic drugs such as acetaminophen, are applied to the patients with influenza viruses as symptomatic treatment. It is very difficult to suppress widespread of influenza viruses without the isolation of patients within 1 or 2 weeks. On the next step of the treatment for influenza, amantadine and rimantadine were administered to patients with influenza A, and ribavirin was used to treat immunosuppressed patients with severe influenza conditions [10]. These are limited in operating only against influenza A viruses and adverse effect and resistance of virus to drugs lead to less use. On the use of these drugs, antiviral effectiveness for alleviation of severe symptoms of patients with seasonal influenza viruses was limited and not enough to suppress the widespread of those? On the third step of the treatment for influenza, NAIs are administered to the patients with influenza viruses. The NA protein is a homotetrameric glycoprotein with a stalk region and enzymatically active head. The NA active site cleaves sialic acid at the glycol-sialic bond on the host cell as well as in respiratory mucus, leading to spread of the virus [11–13]. NAIs act to inhibit the release of progeny viral particles from infected host cells and have more effectiveness and less adverse effects than amantadine and rimantadine when administered to patients with influenza viral infection [14]. Administration of NAIs to patients is recommended within 48 h from the onset of infection [14]. NAIs alleviate symptoms and shorten its duration bothered from typical symptoms of influenza, especially high fever and headache without using antipyretic agents. Four NAIs, namely oseltamivir, zanamivir, laninavir, and peramivir are available in various countries and three measures of administration, namely oral intake, inhalation, and infusion to vein are used [14]. Zanamivir was the first NAI to be developed and was licensed in 1999. Its feature is poor absorption and an inhaled agent and is available in an intravenous form for compassionate use [15]. Oseltamivir is a prodrug being developed on the basis of the structure

adequate vaccination.

3.1. Antiviral therapy

3. Therapy for patients with influenza viruses

#### 2. Prevention

It is not too enough to exaggerate that prevention is the most effective therapy for infectious disease. It is desirable to establish universal most effective vaccine against influenza virus. Vaccine effectiveness (VE) is influenced by viral subtype/lineage as well as the timing of vaccination (early or late epidemic in a season). VE of trivalent influenza vaccine (TIV) is assessed as from 20 to 50% in vaccine programs of several countries [6, 7]. The population rate of people was over 80% in the Korean national immunization program but VE remains low in the elderly adults [8]. They addressed the improvement of influenza vaccine including the adoption of quadrivalent influenza vaccine (QIV), adjuvanted influenza vaccine, and high-dose influenza vaccine. Recently, WHO has been recommending QIV in 2013 and QIV has been adopted in several countries. QIV is estimated cost-effective and cost saving to reduce the burden of outpatient visit for influenza but 1.62 hospitalizations and 0.078 deaths per 100,000 individuals were estimated in Japan [9]. VE is often influenced by the timing between vaccination and influenza epidemic. Early epidemic occurs seasonally before administration of vaccine against seasonal influenza infection in people and VE is low. A mismatch of influenza viral strains between vaccination and epidemic makes VE low. It is very difficult to overcome influenza infection only by vaccination because of current VE and variability of influenza virus. We need the adequate diagnosis and treatment systematically after the procedure of adequate vaccination.


**1. Introduction**

Chilean housing policy has been known as a successful program [1]. Throughout its history, it has positioned itself as a solution to urban informality, being the main problem identified in Chilean cities since the beginning of the twentieth century [2]. From that moment on, the state was characterized by acting reactively to reduce the housing deficit and, in the same way, resolved the precarious living conditions of the people [2, 3]. However, contrary to the housing scheme developed through the first decades of the twentieth century, with the beginning of the military dictatorship in 1973, there was a turn toward a state with neoliberal foundations that understands housing as a commodity and a financial asset [4, 5]. This allowed privatization of social housing production and a consecutive promotion of housing solutions on the private market, meanwhile the state subsidizes the whole process and is responsible for selecting possible inhabitants [6, 7].

Despite this, the current construction of social housing has been reduced, while private housing units have tended to become more expensive, leading to a housing deficit calculated at around 641,000 units in 2022 [8, 9]. Its calculation includes the existence of housing in unlivable structural conditions, overcrowded homes, homeless people, and the growth of informal settlements [9]. The Chilean north is where the construction of informal settlements has had an accelerated and significant growth, concentrating 27% of settlements nationwide [10]. In this region, a complex contradiction is observed, since it is an extractive mining territory that contributes considerably to the economic growth of Chile and its gross domestic product [11]. Mining in connection with unregulated urban planning has promoted unequal and segregated cities, especially reflected in the value of land and access to housing for its population [12, 13] This is explained by the usual mobility of mining workers, whose origin from different parts of the country has driven processes of urban expansion due to real estate speculation [14, 15]. An example of this is the consolidated construction of high-standard housing in the most expensive areas of cities versus the emergence of informal territories, which includes the informal rental and sublease market in central spaces, in addition to informal settlements [12, 16, 17].

In this context, nearly 22,800 families have taken the action of occupying vacant land for the purpose of self-building homes in northern Chile [10]. Among them, there is Chilean population and immigrants from the Latin American and Caribbean region, who settle in peripheral areas within the cities. Although the informal territories can settle in any sector of the cities, their location in disaster-risk areas is recurrent [18, 19]. However, its informality condition has resulted in authorities excluding them from socio-natural disaster prevention programs, leading to an endless accumulation of exposure on its inhabitants [20, 21]. Bringing housing to the center of the disaster debate as reference [19] suggests, we propose mobile housing as an initial proposal to reduce disaster risks that affect informal territories, contributing with suggestions that go beyond the idea of just evicting. More than a housing typology with certain construction characteristics, mobile housing contributes with a set of arguments that question housing policies, governmental reaction to urban informality, and its view on people's residential trajectories. This proposal is based on long-term development of mixed methodologies, started in 2014, that has allowed us a comprehensive understanding of access to housing. It includes the analysis of quantitative data, such as demography, housing, overcrowding, land prices, purchase and sale prices of homes, and rental and sublet prices. Also includes the application of qualitative methods, such as 200 in-depth interviews with residents of the northern cities of Arica, Iquique, Alto Hospicio, Antofagasta, Calama, and San Pedro de Atacama, the development of housing evaluation sheets designed to investigate the architectonic background of informal housing, and the development of meetings and workshops with communities living in informal settlements to learn about their trajectories, views, and opinions.


*2.3. Promoter Isolation and Analysis*

The promoter region of the *Cppsy* gene was cloned from *C. protothecoides* CS-41 genomic DNA. The cloned *Cppsy* promoter region was determined to be 1980 bp in length, and the sequence is shown in Figure 4. Furthermore, the cloned *Cppsy* promoter region was analyzed using the PLACE and PlantCARE databases. Several core fragments were identified, which are homologous to the *cis*-acting elements of higher plants and of great importance for the promoter functions (Figure 4). Three types of elements, which have been found to be regulated by hormones in the upstream region of some

plant genes, are present in the *Cppsy* promoter: the ABRE type (CCTGCGTGGC, CACGTG, and GCCTCGTGGC) involved in abscisic acid responsiveness; the TGACG-motif (TGACG), the *cis*-acting regulatory element involved in methyl jasmonate (MeJA) responsiveness; and the Sp1 (CCCCCGCCA and ACCCGCCATG), MNF (GTGCCCCATGCAGGTT) and Box I (TTTCAAA) types involved in light responsiveness.

The transcriptional start site (TSS) of the *Cppsy* promoter was determined by 5ȝ-RACE using total RNA extracted from *C. protothecoides*. The TSS is an adenine (A) at 34 bp upstream of the translation initiation codon. The distance between the putative  TATA-box and TSS is approximately ƺ24 to ƺ28 bp, which is consistent with the distance of 32 bp ± 7 bp from previous data [30].

A previous study showed that the *psy* expression level is affected by light [31] and other biotic and abiotic stresses [16] in higher plants. Gene expression response to environmental stress is related to the regulation mechanism. To understand more about the regulation mechanism, we need to know more information about the gene, including the gene structure and regulatory domains. Here, many elements were found in the *Cppsy* promoter that belong to light-responsive elements, such as Sp1, MNF1, G-box, and chs-CMA2a. There were also some *cis*-acting elements involved in abscisic acid (ABRE) and MeJA (TGACG-motif) responsiveness. Light is one of the most important environmental factors for algae. To determine whether these motif sequences from the *Cppsy* promoter are involved in light, abscisic acid, and MeJA signaling, loss-of-function analysis needs to be carried out in future studies.

**Figure 4.** Promoter sequence of *Cppsy* from *C. protothecoides* CS-41. Numbers indicate the positions relative to transcriptional start site (TSS). The TSS is indicated as +1 and in bold; important *cis*-elements are underlined.


**4.1. Decline of brook trout**

We sampled 78 streams that historically contained brook trout populations, but found the species in only 56 of them [30]. The range of brook trout is shrinking [39] for several reasons, including habitat alteration, overexploitation, competition with introduced rainbow trout (*O. mykiss*) and brown trout (*S. trutta*) and more recently, climate change.

#### **4.2. Duplicated isozyme loci in brook trout**

Certain allozyme markers posed complications to interpretation of underlying genotype. Brook trout show a high incidence of duplicated enzyme loci due to the tetraploid ancestry of salmonids [40]. Duplicated loci (termed isoloci) are genetically independent, but exhibit alleles of similar electrophoretic mobility that cannot be unambiguously assigned to either locus. Three of the five enzymes that we screened were encoded by isoloci (i.e., *MDH-B1,2*\*, *sAAT-1,2*\*, and *GPI-B1,2*\*). Ambiguous interpretation of the banding patterns of two of these isoloci, *sAAT-1,2*\* and *GPI-B1,2*\*, led us to eliminate them from statistical analysis [30]. Precise estimation of genetic diversity and differentiation metrics require data from many loci [41, 42]. Information from only four markers clearly limited the power of statistical analysis of genetic differentiation, especially with small sample sizes for some of the populations [43]. Genotypic data from more markers likely would reveal genetic differentiation not detected with only four loci. Ongoing screening of additional, more highly polymorphic markers, such as microsatellite DNA markers, will increase the ability to quantify population genetic differentiation.

#### **4.3. Geographic distribution of SABT in southwest Virginia**

Based on fixation for the diagnostic allele at the *CK-A2*\* locus and allele frequency differences at three other marker loci, 34% (*n* = 19) of the brook trout populations analyzed in this study were of southern Appalachian origin, 9% (*n* = 5) were of northern origin, and 57% (*n* = 32) were of mixed genetic origin (**Tables 1** and **2**). The level of certainty for precise characterization of a population is directly related to sample size. That is, any population observed to be fixed for the common allele actually may harbor the alternate allele at a low, undetected frequency. For example, with a sample size (*s*) of 20, our likelihood (*p*) of detecting an allele with a frequency (*p*a ) of 5% is 36% (i.e., *p* = (1−*p*a ) *s* = 0.9520, [44]). Therefore, there is a non-zero likelihood that some populations characterized as "pure" southern Appalachian are of mixed genetic origin. Similarly, sample size also affects estimation of within-population diversity statistics such as *P* and *H*0 . Sampling of a limited number of populations in a watershed also would affect estimates of between-population genetic variability.

Of the six populations from the Holston drainage, four were of mixed genetic origin, with the southern allele at frequencies ranging from 0.44 to 0.95. The Grassy Branch population was characterized as southern Appalachian, and the Henshew Branch population was characterized as pure northern. Results from earlier genetic studies [8, 11, 14] and its geographic location suggest that the Holston River historically contained the southern Appalachian lineage, so the presence of the northern allele is likely due to stocking.

The Yadkin (upper Pee Dee) River is an Atlantic-slope watershed. Despite the common presumption that Atlantic-slope drainages would contain native northern-form brook trout [8, 12, 15], two pure southern Appalachian populations (Pauls Creek, South Fork Stewarts Creek) were found in the Yadkin drainage. Although no early sampling efforts are known from the upper Pee Dee in Virginia [45], the section of the river that flows through North Carolina was excluded from the range of brook trout originally described by Smith [46]. However, several stream capture events have been inferred in this region, suggesting that these populations are descendants of brook trout captured from the New River [45]. Inspection of stocking records showed that both Pauls Creek and South Fork Stewarts Creek were stocked in the recent past, implying that the "native" southern strain persisted despite stocking.

Earlier genetic study [14] and geographic location suggest that the James River historically contained northern-form brook trout. Three populations from the James River screened in this study were characterized as northern form. This finding leaves little doubt that the New River is the boundary between northern and southern Appalachian brook trout populations.

In this study, 16 populations from the New River drainage (36%) were characterized as southern Appalachian brook trout. No geographic patterns of genetic variation were observed among the populations of putative pure southern origin. Interestingly, two of these "pure southern" populations (Crooked Creek and West Fork Dry Run) were stocked in the recent past with northern-derived hatchery fish. Crooked Creek is a "put-and-take" fishing area, and 5000 brook trout are stocked annually, yet it maintained an apparently pure southern population. Sixty-three percent of the populations from the New River drainage were of mixed origin, with the southern allele at frequencies ranging from 0.21 to 0.98. Although stocking records are limited, only two of these (Howell Creek and Little Indian Creek) are known to have been stocked with northern-derived hatchery fish. Only one population (Pearis Thompson Branch) in the New River was characterized as pure northern.

In addition to the 56 populations characterized in this study, we compiled data from all known genetic studies of brook trout populations in southwest Virginia [12, 14, 15]. Fortyseven percent (*n* = 39) of all 83 populations characterized in southwest Virginia were of mixed genetic origin (**Table 3**); however, many of these introgressed populations were largely southern. In addition, the "pure" southern populations (*n* = 26) that remain provide opportunities for restoration of southern Appalachian brook trout in Virginia.

### **4.4. Range-wide geographic distribution and genetic affinity of New River brook trout populations**

With the zone of contact between the northern and southern forms lying roughly at the New River watershed, it is unknown whether the New River historically contained the pure southern Appalachian form, or whether it was a zone of intergradation among southern and northern Appalachian lineages. Interpreting data across this study and the three studies noted above [12, 14, 15], the New River drainage contains 20 pure southern populations, suggesting that the presence of northern alleles could be due to either stocking or stream capture events. However, a large proportion (64%) of populations from the New

River are of mixed genetic origin, suggesting either that hatchery fish persisted in the New watershed or that the New River is a zone of natural intergradation. To gain a better understanding of the geographic distribution of southern Appalachian brook trout, we compiled

**Table 3.** Genetic characterization at the *CK-A2\** locus for southwest Virginia brook trout populations not sampled in this

*N* = number of individuals per sample.

study, compiled from both published and unpublished data sources.

**Stream River drainage County** *N* **% Southern allele Source** Green Cove Creek Holston Washington 19 95 [15] Grindstone Branch Holston Smyth 16 97 [14] Houndshell Branch Holston Smyth 12 100 [14] Jerry Creek Holston Smyth 11 100 [14] Little Laurel Creek Holston Smyth 16 100 [14] Johns Creek James Giles 23 0 [14] Shawvers Run James Giles 23 0 [14] Spy Run James Augusta 21 0 [14] Valley Branch James Craig 15 0 [14] Burks Fork New Floyd 15 67 [15] Cox Branch New Tazewell 15 53 [15] Dry Creek New Smyth 24 100 [14] Hanks/EF Chestnut Creek New Grayson 10 70 [14] Helton Creek New Grayson 21 79 [15] Jerry Creek New Grayson 15 67 [15] Killinger Creek New Smyth 12 88 [14] Laurel Branch New Floyd 15 97 [14] Laurel Fork New Floyd 7 79 [12] Lewis Fork New Grayson 21 79 [15] Middle Fork Helton New Grayson 20 100 [14] NF Elk Creek New Grayson 19 100 [14] NP Buckhorn Creek New Carroll 25 100 [14] Wilburn Branch New Grayson 21 75 [15] Big Stony Creek Roanoke Bedford 10 0 [12] Little Stony Creek Roanoke Bedford 6 0 [12] Rock Castle Creek Roanoke Patrick 25 36 [14] Turkey Creek Yadkin Carroll 15 47 [15]

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65

Genetic Characteristics of Southern and Northern Brook Trout (*Salvelinus fontinalis*) Populations... http://dx.doi.org/10.5772/intechopen.70719 65

The Yadkin (upper Pee Dee) River is an Atlantic-slope watershed. Despite the common presumption that Atlantic-slope drainages would contain native northern-form brook trout [8, 12, 15], two pure southern Appalachian populations (Pauls Creek, South Fork Stewarts Creek) were found in the Yadkin drainage. Although no early sampling efforts are known from the upper Pee Dee in Virginia [45], the section of the river that flows through North Carolina was excluded from the range of brook trout originally described by Smith [46]. However, several stream capture events have been inferred in this region, suggesting that these populations are descendants of brook trout captured from the New River [45]. Inspection of stocking records showed that both Pauls Creek and South Fork Stewarts Creek were stocked in the recent past, implying that the "native" southern strain persisted despite stocking.

Earlier genetic study [14] and geographic location suggest that the James River historically contained northern-form brook trout. Three populations from the James River screened in this study were characterized as northern form. This finding leaves little doubt that the New River is the boundary between northern and southern Appalachian brook trout populations.

In this study, 16 populations from the New River drainage (36%) were characterized as southern Appalachian brook trout. No geographic patterns of genetic variation were observed among the populations of putative pure southern origin. Interestingly, two of these "pure southern" populations (Crooked Creek and West Fork Dry Run) were stocked in the recent past with northern-derived hatchery fish. Crooked Creek is a "put-and-take" fishing area, and 5000 brook trout are stocked annually, yet it maintained an apparently pure southern population. Sixty-three percent of the populations from the New River drainage were of mixed origin, with the southern allele at frequencies ranging from 0.21 to 0.98. Although stocking records are limited, only two of these (Howell Creek and Little Indian Creek) are known to have been stocked with northern-derived hatchery fish. Only one population (Pearis Thompson Branch)

In addition to the 56 populations characterized in this study, we compiled data from all known genetic studies of brook trout populations in southwest Virginia [12, 14, 15]. Fortyseven percent (*n* = 39) of all 83 populations characterized in southwest Virginia were of mixed genetic origin (**Table 3**); however, many of these introgressed populations were largely southern. In addition, the "pure" southern populations (*n* = 26) that remain provide opportunities

**4.4. Range-wide geographic distribution and genetic affinity of New River brook trout**

With the zone of contact between the northern and southern forms lying roughly at the New River watershed, it is unknown whether the New River historically contained the pure southern Appalachian form, or whether it was a zone of intergradation among southern and northern Appalachian lineages. Interpreting data across this study and the three studies noted above [12, 14, 15], the New River drainage contains 20 pure southern populations, suggesting that the presence of northern alleles could be due to either stocking or stream capture events. However, a large proportion (64%) of populations from the New

in the New River was characterized as pure northern.

**populations**

64 Biological Resources of Water

for restoration of southern Appalachian brook trout in Virginia.

**Table 3.** Genetic characterization at the *CK-A2\** locus for southwest Virginia brook trout populations not sampled in this study, compiled from both published and unpublished data sources.

River are of mixed genetic origin, suggesting either that hatchery fish persisted in the New watershed or that the New River is a zone of natural intergradation. To gain a better understanding of the geographic distribution of southern Appalachian brook trout, we compiled allele frequency data from all known genetic studies of brook trout populations throughout the native range (**Table 4**). Frequencies of the *CK-A2*\*100 (i.e., southern) allele were weighted based on sample size and averaged across all populations in each river drainage. **Figure 2** shows the frequency of the southern allele in each of the major river drainages from which data were collected.

All river drainages north of the New River were characterized as pure northern, with the exception of the Roanoke River drainage that contained a single population with a low frequency of the southern allele, likely due to the transfer of individuals from another location or stream capture. The frequency of the southern allele in river drainages south of the New River ranges from 29% in the Broad River of North Carolina to 100% in the Coosa River of Georgia. Genetic characterization of individuals from 111 populations in the New River drainage showed an 85% frequency of the southern-form allele. **Figure 2** shows that the south/north break is sharp and that this break occurs at the New/Roanoke-James watershed divide. This weakens the hypothesis that the New River is a zone of natural intergradation between the southern and northern forms of brook trout, and supports the hypothesis that the presence of northern alleles is due to either stocking or stream capture. However, it is important to qualify this inference by noting that genetic characterization is based on variation at a single locus. Ongoing screening of New River populations using microsatellite DNA markers will provide further insights into patterns of population genetic differentiation, shedding light on the native character of New River brook trout populations. In particular, microsatellite variation may clarify whether northern alleles observed in populations examined are characteristic

**Figure 2.** Genetic characterization of brook trout populations in major river drainages, based on the *CK-A2*\* locus, using data compiled from all known genetic studies of brook trout populations throughout the native range. See **Table 4** for

Northern Southern Introgressed

Genetic Characterization

WV

Genetic Characteristics of Southern and Northern Brook Trout (*Salvelinus fontinalis*) Populations...

**9**

**14**

**10**

**25**

**13**

River Drainage 1 Susquehanna 15 Savannah 2 Ohio 16 Nolichucky 3 Gunpowder 17 Nantahala 4 Patapsco 18 Tuckasegee 5 Potomac 19 French Broad 6 James 20 Hiwassee 7 Rappahannock 21 Cheoah 8 Roanoke 22 Little 9 New 23 Pigeon 10 Broad 24 Little Tennessee 11 Chattahoochee 25 Watauga 12 Tellico 26 Holston 13 Catawba 27 Tennessee 14 Yadkin 28 Coosa

MD

**4**

**1**

67

PA

**2 3**

http://dx.doi.org/10.5772/intechopen.70719

**5**

VA

**7**

**6**

**8**

of particular hatchery stocks or of native regional variation.

0 50 0 50000<PHONE_NUMBER>00 150000200 25<PHONE_NUMBER>000 kilometers

OH

NC TN

**19**

**18**

**16**

**26**

**15**

GA SC

**17**

**22 23**

**11**

**20**

**12**

**21**

**24**

KY

**28**

details.

**27**

N

1 Relative to eastern continental divide.

2 Allele frequency based on number of individuals analyzed per stream and averaged across all populations in each drainage.

**Table 4.** Genetic characterization of brook trout populations in regional river drainages, based on frequency of the diagnostic *CK-A2\*100* allele using data gathered from all available published and unpublished studies.

Genetic Characteristics of Southern and Northern Brook Trout (*Salvelinus fontinalis*) Populations... http://dx.doi.org/10.5772/intechopen.70719 67

allele frequency data from all known genetic studies of brook trout populations throughout the native range (**Table 4**). Frequencies of the *CK-A2*\*100 (i.e., southern) allele were weighted based on sample size and averaged across all populations in each river drainage. **Figure 2** shows the frequency of the southern allele in each of the major river drainages

**River drainage State Position1 # of streams # of individuals % Southern2 Source(s)** Susquehanna PA/MD East 4 145 0 [7, 9] Ohio MD West 3 110 0 [9] Gunpowder MD East 1 40 0 [9] Patapsco MD East 1 40 0 [9] Potomac MD/VA East 6 190 0 [9, 14] James VA East 7 142 0 [14, current]

Rappahannock VA East 1 25 0 [14] Roanoke VA East 3 41 22 [12, 14] New VA/NC West 101 1999 85 [14,

Yadkin VA/NC East 37 691 58 [8, 12,

Nolichucky NC/TN West 51 1058 64 [7, 8, 11] French Broad NC/TN West 80 1281 73 [8, 11, 16] Little Tennessee NC/TN West 49 886 82 [8, 13] Watauga NC/TN West 44 691 88 [8, 11] Broad NC East 3 41 29 [8, 11] Hiwassee NC West 6 146 76 [8, 11] Cheoah NC West 10 210 80 [8, 11] Little TN West 8 90 80 [8, 11] Tellico TN West 5 64 42 [11] Savannah NC/GA East 27 533 63 [10, 16] Chattahoochee GA West 1 21 31 [10] Tennessee GA West 7 145 93 [10] Coosa GA West 1 12 100 [10]

Holston VA/TN West 24 320 91 [8, 11, 14, current]

Allele frequency based on number of individuals analyzed per stream and averaged across all populations in each

**Table 4.** Genetic characterization of brook trout populations in regional river drainages, based on frequency of the

diagnostic *CK-A2\*100* allele using data gathered from all available published and unpublished studies.

15, current]

15, current]

from which data were collected.

66 Biological Resources of Water

1

2

drainage.

Relative to eastern continental divide.

**Figure 2.** Genetic characterization of brook trout populations in major river drainages, based on the *CK-A2*\* locus, using data compiled from all known genetic studies of brook trout populations throughout the native range. See **Table 4** for details.

All river drainages north of the New River were characterized as pure northern, with the exception of the Roanoke River drainage that contained a single population with a low frequency of the southern allele, likely due to the transfer of individuals from another location or stream capture. The frequency of the southern allele in river drainages south of the New River ranges from 29% in the Broad River of North Carolina to 100% in the Coosa River of Georgia. Genetic characterization of individuals from 111 populations in the New River drainage showed an 85% frequency of the southern-form allele. **Figure 2** shows that the south/north break is sharp and that this break occurs at the New/Roanoke-James watershed divide. This weakens the hypothesis that the New River is a zone of natural intergradation between the southern and northern forms of brook trout, and supports the hypothesis that the presence of northern alleles is due to either stocking or stream capture. However, it is important to qualify this inference by noting that genetic characterization is based on variation at a single locus. Ongoing screening of New River populations using microsatellite DNA markers will provide further insights into patterns of population genetic differentiation, shedding light on the native character of New River brook trout populations. In particular, microsatellite variation may clarify whether northern alleles observed in populations examined are characteristic of particular hatchery stocks or of native regional variation.

#### **4.5. Management implications**

Brook trout is the only salmonid native to the southern Appalachian region. The American Fisheries Society Southern Division Trout Committee developed a position statement [22] expressing the importance of SABT and presenting recommendations for conservation-oriented management of this regional resource. Our results contribute to the recommended completion of genetic inventory of critical populations using non-lethal sampling methods. In this context, we frame the management implications for management of SABT populations.

knoxnews.com/news/aquarium-helping-to-restore-native-trout-ep-510367109-355447741. html). SABT can be stocked to re-establish populations in streams where they have been extirpated. Also, while we do not recommend eradicating non-native or introgressed populations in watersheds where brook trout are native, we recommend stocking southern-strain hatchery fish into these populations to shift allele frequencies toward those of native populations. Progress in re-establishing native brook trout populations should be monitored using genetic

Genetic Characteristics of Southern and Northern Brook Trout (*Salvelinus fontinalis*) Populations...

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69

This work was funded through the Federal Aid in Sport Fish Restoration Project F-128-R, administered by the Virginia Department of Game and Inland Fisheries, and is based on the Master's degree research of Joanne (Davis) Printz. We thank George Palmer and Cliff Kirk for assistance with fieldwork and collection of genetic samples. Ray Morgan of the Maryland Department of Natural Resources kindly provided unpublished data from Maryland populations, and Doug Besler of the NCWRC generously provided tissue samples. Finally, we thank Chris Printz of ATS International, Inc. of Christiansburg, VA for his assistance in the design and production of the maps. Funding for EH's participation in this work was provided in part by the Virginia Agricultural Experiment Station and the Hatch Program of the National

and Eric M. Hallerman1

1 Department of Fish and Wildlife Conservation, Virginia Polytechnic Institute and State

[1] MacCrimmon HR, Campbell JS. World distribution of brook trout, *Salvelinus fontinalis*. Journal of the Fisheries Research Board of Canada. 1969;**26**:1699-1723. DOI: 10.1139/

[2] Hudy M, Thieling TM, Gillespie N, Smith EP. Distribution, status, and land use characteristics of subwatersheds within the native range of brook trout in the eastern United States. North American Journal of Fisheries Management. 2008;28:1069-1085. DOI: http://

2 Virginia Department of Game and Inland Fisheries, Blacksburg, VA, USA

\*

Institute of Food and Agriculture, U.S. Department of Agriculture.

, Joseph Williams2

\*Address all correspondence to<EMAIL_ADDRESS>
University, Blacksburg, VA, USA

dx.doi.org/10.1577/M07-017.1.

markers every few generations.

**Acknowledgements**

**Author details**

Joanne E. Printz1

**References**

f69-159

Results from this and other studies demonstrate that stocking of non-native genotypes poses long-term genetic impacts and interferes with efforts to conserve southern Appalachian brook trout. Although negative effects of stocking have become well known, some fisheries management agencies maintain imprecise stocking records. Further, hatchery personnel often substitute one brook trout stock for another based on availability. We recommend that all stocking and transfers of brook trout be well planned with cognizance of genetic conservation objectives and thoroughly and accurately documented.

Management units—that is, populations that are demographically independent of one another—may be defined functionally as populations that have substantially divergent allele frequencies at many loci [47]. We had but limited ability to estimate levels of genetic diversity and differentiation among regional brook trout populations using allozyme markers. The results of ongoing screening of microsatellite DNA markers will be used to quantify differentiation among native populations, providing the basis for defining defensible management units. Results to date support the view that southern Appalachian brook trout populations should be managed on a stream-by-stream basis.

Those populations characterized as pure SABT should be given conservation priority. The stocking and transfer of non-native genotypes into these populations should be prohibited. Harvest should be allowed only in those populations that are demographically able to sustain themselves. We recommend that introgressed populations that contain less than 5% admixture from northern-strain brook trout be treated as 'pure' southern. However, we caution that the level of introgression in these populations may be higher than allozyme frequencies suggest; hence, individuals from these streams should not be transferred into streams that contain pure SABT populations. Hatchery brook trout should be stocked only into those streams that contain pure northern-strain populations and those with greater than 5% admixture.

We caution that any negative consequences of stocking also would apply to native northernstrain populations (i.e., in the James and Roanoke river drainages). Allozyme markers do not provide enough resolution to differentiate between native northern and hatchery populations, and so we recommend that all brook trout populations should be screened and characterized using microsatellite or single nucleotide polymorphism markers. Until we know more about the genetic composition of these populations, it may be wise to stock only infertile triploid brook trout [48].

Southern Appalachian brook trout hatchery stocks are being established in conservationoriented hatchery programs ([49], https://brooktrouthatchery.wordpress.com/, http://archive. knoxnews.com/news/aquarium-helping-to-restore-native-trout-ep-510367109-355447741. html). SABT can be stocked to re-establish populations in streams where they have been extirpated. Also, while we do not recommend eradicating non-native or introgressed populations in watersheds where brook trout are native, we recommend stocking southern-strain hatchery fish into these populations to shift allele frequencies toward those of native populations. Progress in re-establishing native brook trout populations should be monitored using genetic markers every few generations.


**Abstract**

Polypyridyl ruthenium(II) complexes have been widely researched as promising functional molecules. We have found unique photoisomerization reactions of polypyridyl ruthenium(II) aquo complexes. Recently we have attempted to provide insight into the mechanism of the photoisomerization of the complexes and distinguish between the *distal*�/*proximal*-isomers in their physicochemical properties and functions. Moreover, polypyridyl ruthenium(II) aquo complexes have been intensively studied as active water oxidation catalysts (WOCs) which are indispensable for artificial photosynthesis. The catalytic aspect and mechanism of water oxidation by the *distal-/proximal*-isomers of polypyridyl ruthenium(II) aquo complexes have been investigated to provide the guided thought to develop more efficient molecular catalysts for water oxidation. The recent progress on the photoisomerization and water oxidation of polypyridyl ruthenium(II) aquo complexes in our group are reviewed to understand the properties and functions of ruthenium complexes.

**Keywords:** Ruthenium aquo complexes, Photoisomerization, Water oxidation catalysis, Artificial photosynthesis

## **1. Introduction**

Polypyridyl ruthenium(II) complexes have been widely researched as promising functional molecules due to appealing photochemical [1–3] and photophysical [4–6] properties as well as redox properties [7, 8], which enable them to exhibit a number of functions such as electrochromism [9, 10], proton-coupled electron transfer [11–13] and photocatalysis [14, 15]. As a result, the polypyridyl ruthenium(II) complexes have been applied to a large variety of devices including sensors [16], photovoltaic cells [17], displays [18] and artificial photosynthesis [19, 20].

We presented irreversible and stoichiometric photoisomerization of *distal*-[Ru (tpy)(pynp)OH2] 2+ (*d*-**1**) (tpy = 2,20 ;60 ,2″-terpyridine, pynp = 2-(2-pyridyl)-1,8 naphthyridine) to *proximal*-[Ru(tpy)(pynp)OH2] 2+ (*p*-**1**) as shown in **Figure 1** [21–23], which had not been characterized previously for polypyridyl ruthenium

**Figure 1.**

*Photoisomerization of* distal-*[Ru(Rtpy)(pynp)OH2] 2+ to* proximal-*isomers.*

(II) aquo complexes although various photochemical reactions of the ruthenium(II) complexes have been reported [24–33]. We have attempted to provide insight into the mechanism of the photoisomerization of polypyridyl ruthenium(II) aquo complexes and distinguish between the *distal*�/*proximal*-isomers in their physicochemical properties and functions [21–23, 34–36]. We have also developed new synthetic strategy to form dinuclear ruthenium(II) complexes utilizing the photoisomerization [37, 38]. Moreover, polypyridyl ruthenium(II) aquo complexes have been intensively studied as active WOCs [21, 22, 34, 35, 37, 39, 40] which are indispensable for artificial photosynthesis. The catalytic aspect and mechanism of water oxidation by the *distal*-/*proximal*-isomers of polypyridyl ruthenium(II) aquo complexes have been investigated to provide the guided thought to develop more efficient molecular catalysts for water oxidation. In this chapter, we review the recent progress on the photoisomerization and water oxidation of polypyridyl ruthenium(II) aquo complexes in our group.


**3. Results and discussion**

#### **3.1 TAR model results**

LLCUL\_CAP is the threshold variable and is separated into two regimes with observations. The TAR model shows the threshold value for both regimes to be 2.09%, with the first regime consisting of 16 observations (*Zt*1 < *λ*) and the second regime having 22 observations (*Zt*1>*λ*). The threshold level is negative in the analysis because arable land is a fixed asset that cannot be increased, while the South African population growth is increasing. The relationship between the threshold variables and the explanatory variables (price per ton and consumption per ton) is differentiated into the different regimes. In the first regime with 16 observations, the price per ton shows a coefficient of 0.1365 and carries a negatively related coefficient against the cultivated land/area, which means that an increase in LPRCE\_TON has the effect of decreasing LLCUL\_CAP (per capita maize-cultivated land) with a margin of 13.7%; this is shown in **Table 1**.

Consumption per ton (LCONSUPTION\_TON) also carries a negatively related coefficient of 0.8236 against the threshold variable (cultivated land) which suggests that increases in consumption per ton put downwards production pressure on cultivated land area, with a margin of 82.4%. The denoted relationship therefore resulted in 16 observations that are less than the threshold value which suggests that during the pre-1994 apartheid era, the negative relationship was sustainable and did not go past the threshold level of 2.09%; in the first regime, the LPRCE\_TON is not statistically significant with t-statistics and p-value that are above the 5% significance level, while LCONSUPTION\_TON is reported to be statistically significant as p-values are less than 5%; this is shown in **Table 1**.

**Threshold**

**LLCUL\_CAP(**

**29**

**3) < 2.09 with 16**

Zt**1** <λ **(first regime)**

**Coefficient**

0.136533

0.823665

Non-threshold

5.621432

2.146702

 2.618636

0.0132

 variable

 0.280937

 0.085868

LPRCE\_TON

LCONSUPTION\_TON

C **Table 1.** *TAR model estimates.*

 **Std. error**

 **t-statistics** 1.590037

2.931847

 0.0061

 0.1214

 **p-values**

 **Coefficient**

0.265049

0.690729

 0.257767

 0.042998

 **Std. error**

 **t-statistics** 6.164189

2.679666

 0.0114

 0.0000

 **p-values**

*Regime Switch and Effect on Per Capita Food Security Issues in South Africa*

*DOI: http://dx.doi.org/10.5772/intechopen.86931*

**observations**

 **variables** **LLCUL\_CAP(**

**3) = > 2.09 with 22**

Zt**1**>λ

**(second regime)**

**observations**

In the case of the second regime, both explanatory variables are statistically significant, with p-values that are below 5%. The threshold variable (cultivated land) has 22 observations that are below the threshold value indicating unsustainable per capita food security. The coefficient for both price per ton and consumption per ton is still negatively related in the regime, with LPRCE\_TON shown as 0.265 and LCONSUPTION\_TON as 0.6907. These values suggest that an increase in LPRCE\_TON and LCONSUPTION\_TON results in decreasing cultivated area/land of maize. The results suggest the cultivated land area of maize was not sustainable at the threshold value of 2.09% for food security and imply that the cultivated land area has diminished beyond the negative threshold value with at least 22 observations.

A more descriptive observation of the result is shown by **Figure 1** for actual, fitted and residual model for the transformed data. More specifically, attention is given to the actual and fitted model in the figure. As from 1970 to 1988, the first regime is observed, while the second regime continues from 1989 to 2010, where the first regime with 16 observations shows the cultivated land remains sustainable even though it negatively slopes and decreases. However, in the second regime, as located from 1989, the behaviour changes, because the actual and fitted models show a further decline in cultivated land area beyond the threshold value at 22 observations; this makes the second regime unsustainable.

The results of per capita consumption and the change in consumption trend over the period of analysis from 1970 to 2010 are indicated in **Figure 1**. The food security indicators (per capita land cultivated and per capita consumption) presented in **Figure 1** indicate the validity of food sovereignty theory with the evidence that

*TAR model estimates.*


*3.4. Dropout*

Neural networks are prone to overfitting. Overfitting is essentially the exhaustive training of the model in a certain set of data, so much that the model fails to generalize. As a result, the model cannot effectively work with new unknown data. A solution to overfitting is to train multiple models and combine their output afterwards, which is highly inefficient.

Srivastava et al. [**?** ] proposed randomly dropping neural units from the network during the training phase. Their results suggested an improvement of the regularization on diverse datasets. Spatial dropout refers to the exact same process, performed over a single axis of elements, rather than random neural units over each layer. Furthermore, dropout has a significant potential to reduce overfitting and provide improvements over other regularization strategies such as *L*-regularization and soft-weight sharing [**?** ].

### *3.5. LSTM and Gated Recurrent Unit*

A feed-forward neural network has a unidirectional processing path, from input to hidden layer to output. A recurrent network can have information travelling both directions by using feedback loops. Computations derived from earlier input are fed back into the network, imitating human memory. In essence, a recurrent neural network is a chain of identical neural networks that transfer the derived knowledge from one to another. That chain creates learning dependencies that decay mainly due to the size of the chained network.

Hochreiter and Schmidhuber [**?** ] proposed Long Short-Term Memory networks to counter that decay. The novel unit of the LSTM architecture is the memory cell that forgets or remembers the information passed from the previous chain link. The Gated Recurrent Unit of Model 2 was introduced by Cho et al. [**?** ]. Its architecture is similar to LSTM units, but with the absence of an output gate. It is shown that GRU networks perform well on Natural Language Processing tasks [**?** ].