**4. Conclusions**

A distinct salinity plume was observed around the outfall of Carlsbad Desalination Plant with a salinity increase of up to 2.7 units above ambient and extending 600 m offshore. This exceeds the maximum allowed salinity perturbation in the coastal zone as stated by the California Ocean Plan (2015 Amendment to Water Quality Control Plan). However, no significant impact was found on the benthic epifauna or infaunal composition and abundance in the same area, possibly due to an already disturbed conditions in this setting due to previous and ongoing discharge of post-cooling water from a local power plant. The coastal wave energy and mixing potential at Carlsbad Beach is high compared to other locations along the Southern and Central California coastline, and the lack of sufficient mixing of the brine at Carlsbad Beach raises concern regarding the efficiency of brine mixing at other proposed sites for desalination plants along the coast of California.

We recommend all future desalination facilities to be designed with optimal discharge options of either diffusor systems or a higher dilution of the brine prior to discharge. Furthermore, they should, where possible, be co-located with other water discharging facilities such as power plant or waste water treatment plants to restrict potential impacts to areas that are already disturbed from anthropogenic activities. Finally, we stress the need for long-term monitoring of coastal ecosystems in the area surrounding proposed and current desalination facilities to fully understand the ecological response to long-term exposure to salinity increase.

**Supplementary Materials:** The following are available online at http://www.mdpi.com/2073-4441/11/2/208/s1, Figure S1: Benthic macrofauna. Figure S2: Sediment grain size distribution, Table S1: Water chemical properties. Table S2: Brittle star growth. Table S3: Wave energy and benthic habitat estimation.

**Author Contributions:** Conceptualization, K.L.P., D.P. and A.P.; Data Curation, K.L.P. and A.P.; Formal Analysis, K.L.P., N.H., B.G.R. and A.H.; Funding Acquisition, D.P. and A.P. and K.L.P.; Investigation, K.L.P., D.P. and A.P.; Methodology, N.H. and B.G.R.; Project Administration, A.P. and K.L.P.; Resources, A.P.; Software, N.H. and B.G.R.; Supervision, D.P. and A.P.; Validation, D.P.; Visualization, K.L.P., N.H. and B.G.R.; Writing—Original Draft, K.L.P.; Writing—Review and Editing, K.L.P., N.H., B.G.R., D.P., A.H. and A.P.

**Funding:** This work was supported by National Science Foundation COASTAL SEES #1325649 awarded A.P and D.P. K.L.P. was partially supported by student fellowships from the Geological Society of America (#: 11215-16), the Weigel Scholarship for Coastal Studies, Myers Oceanographic and Marine Trust and the Explorers Club Youth Foundation (#: 83402).

**Acknowledgments:** We would further like to acknowledge the students and researchers of the Paytan Lab at UCSC, in particular Joseph Murray, Ana Martinez Fernandez, Kim Bitterwolf, Kyle Broach and Katie Roberts and the scientific divers and boat crew, especially Jacque Lord and Rich Walsh, for their extensive help and support in completing this work.

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


**5. Conclusions and Discussion**

As the main actors of environmental protection and green governance at the micro level, corporates play a crucial role in achieving sustainability goals. Based on the upper echelons theory, it is believed that executives from environmental protection backgrounds tend to integrate previous "green" cognitive experience into decision-making behavior, ensuring the improvement of their environmental and economic performance. This study focuses on whether listed companies employing executives with environmental protection backgrounds can promote green innovation and its boundary conditions. The study finds that executives with environmental protection backgrounds enhance green innovation. The positive relationship remains robust when instrumental variables and a series of robustness tests address the endogeneity issue. Further, the positive relationship between executives' environmental protection background and green innovation is strengthened when media attention and board independence are greater. The findings provide insights into the view that executive characteristics impact green innovation.

This study extends the literature on the governance effects of management characteristics and provides new empirical evidence for the study of green innovation. First, this study adds value to the literature on upper echelons theory and green innovation research. Prior research on the effects of executive characteristics on green innovation has focused on the effects of characteristics such as executive compensation, educational background, overseas experience, and executive tenure [5]. However, few studies have put the perspective of the executive's environmental protection background affecting green innovation. Therefore, this study extends the executive's environmental protection background to the field of green innovation research based on upper echelons theory. Second, it adds to the work of Khanra et al. (2022) [27] who examined the strategic management actions taken by executives in introducing green innovation initiatives, further adding to this literature by focusing

on how the executive's environmental protection background influences the boundary mechanisms of green innovation. While previous research has focused on the direct effects of executive characteristics on various strategic choices of firms, scholars have recently called for further exploration of factors that mitigate or enhance these effects [6]. Based on insights from agency theory, this study examines the moderating role of media attention and board independence. This study also supplements the previous interpretation of agency theory and introduces the role of internal and external supervision mechanism [68]. This study extends the view that integrating agency theory and upper echelons theory enriches understanding of the governance functions of these two supervisory mechanisms in the executive's environmental protection background.

#### **6. Recommendation**

To enhance corporate green governance and sustainable development, it is important to leverage the governance strengths of executives and add an "environmental barrier" to corporate business decisions. The government should guide companies to set scientific standards for executive staffing and hire executives who value corporate environmental governance, so that business development can balance financial performance with environmental performance. In addition, executives with environmental protection backgrounds should be given the same rights as their positions to ensure they have a sufficient voice in corporate decision-making to promote sustainable development. Based on the research findings, several practical recommendations are made for governments and corporates.

For the government, it must increase its policy support to encourage corporates to implement green innovation. As a way for corporates to take up social and environmental responsibility, green innovation goals are not only based on the realization of their economic benefits, but also take into account the embodiment of social responsibility, urging enterprises to improve environmental performance and enhance green innovation. The government should build a perfect green innovation system, actively guide the flow of green funds to corporates, and prompt them to eliminate their backward production methods to promote environmental protection and sustainable development of the economy. For example, in the context of low-carbon development, the government should introduce policies and measures to promote green innovation to ensure resource efficiency. It could establish a special fund for corporate green finance or credit. It should follow the principle of differentiation and formulate targeted preferential policies, according to the actual situation of corporates, to support the deep integration of green products, organizational structures, and management processes.

For corporates, it is important to emphasize the role of the environmental protection background of executives in enhancing green innovation. First, boards must consider this characteristic when selecting executives and ensuring that their decisions are consistent with corporate goals. For example, suppose corporates are under pressure for environmental legitimacy (e.g., corporate pollution). In that case, executives with environmental protection backgrounds may be an effective way to make the necessary strategic changes [48]. Executives with an environmental protection background are beneficial from an environmental protection perspective, especially in the context of green innovation, an area considered important for corporate strategy. Hiring executives with environmental protection backgrounds in listed companies helps to encourage green investors to invest in such companies. As the number of executives with environmental protection backgrounds increases, their contribution to green innovation becomes more evident. Therefore, corporates should improve their governance mechanisms. Including executives with environmental protection backgrounds in corporate management can improve management diversity, thus reducing the myopia of management and facilitating the management to make green innovation decisions.

Second, in the era of big data, corporates should promote information dissemination and corporate governance through the media, so that stakeholders can better understand the corporate and improve its information transparency. Corporates should raise the environmental protection awareness of their executives and be more active in disclosing green innovation information. This can alleviate the information asymmetry between enterprises and stakeholders and compensate for the lack of contrast resulting from high stakeholder expectations. Corporates can gain the trust and support of stakeholders, thus promoting green innovation. Corporates should promote information dissemination and corporate governance through the media so that stakeholders can improve the transparency of information and social responsibility of corporates.

Third, since the higher the proportion of independent directors, the easier it is for executives with environmental protection backgrounds to play a role and thus promote green innovation, it is necessary to improve the corporate governance mechanism and improve the supervision mechanism on executives' decision-making process, which can be achieved through measures such as increasing the proportion of independent directors and designing an investment risk-sharing mechanism.


*4.5. GC Analysis*

The GC analysis was carried out using an Agilent 6890N GC system Agilent 5975 (SEM Ltd., Istanbul, Turkey). The FID detector temperature was 300 ◦C. To obtain the same elution order with GC-MS, simultaneous auto-injection was done on a duplicate of the same column applying the same operational conditions. Relative percentage amounts of the separated compounds were calculated from FID chromatograms. The analysis results are given in Table 1.

Identification of the essential oil components was carried out by comparison of their relative retention times with those of authentic samples or by comparison of their relative retention index (RRI) to series of *n*-alkanes [25,26]. Computer matching against commercial (Wiley GC/MS Library, MassFinder Software 4.0) and in-house "Ba¸ser Library of Essential Oil Constituents" which includes over 3200 genuine compounds with MS and retention data from pure standard compounds and components of known oils as.

#### *4.6. Insects*

*Aedes aegypti* used in these studies were from a laboratory colony maintained at the Mosquito and Fly Research Unit, Center for Medical, Agricultural and Veterinary Entomology, USDA-ARS, Gainesville, Florida since 1952. We received the eggs and stored them in our laboratory until needed. Mosquitoes were reared to the adult stage by feeding the larvae on a larval diet of 2% slurry of 3:2 beef liver powder (now Foods, Bloomingdale, Illinois) and Brewer's yeast (Lewis Laboratories Ltd., Westport, CT, USA). The eggs were hatched and reared to the pupal stage in an environment-controlled room at a temperature of 27 ◦C ± 2 ◦C and 60 ± 10% RH in a photoperiod regimen of 12:12 (L: D) h. The adults were fed on cotton pads moistened with a 10% sucrose solution placed on the top of screens of 4-L cages.

#### *4.7. Mosquito Biting Bioassay*

Bioassays were conducted using a six-celled in vitro Klun and Debboun (K & D) module bioassay system developed by Klun et al. [27] for quantitative evaluation of biting deterrent properties of compounds. The K & D system consists of a six-well reservoir with each of the 4 × 3 cm wells containing 6 mL of feeding solution. We used the CPDA-1± ATP solution instead of human blood [22]. CPDA-1 and ATP preparations were freshly made on the day of the test and contained a green fluorescent tracer dye (fluorescent water-based tracer "Green"; www.blacklightworld.com) that allowed for the identification of mosquitoes that were fed on the solution. The squashed mosquitoes were observed under black light (FEIT, BPESL15T/BLB 13W 120VAC 60Hz 200mA, Ul#E170906) for feeding. DEET (97% purity *N*,*N*-diethyl-3-methylbenzamide) was used as a positive control (Sigma-Aldrich Co., St. Louis, MO, USA) and ethanol (Fisher Scientific Chemical Co. Fairlawn, NJ, USA) was used as solvent control. Stock and dilutions of all extracts and DEET were prepared in ethanol. All essential oils were evaluated at dosages of 10 μg/cm2 and DEET along with the pure compounds was tested at a concentration of 25 nmol/cm2.

The temperature of the solution was maintained at 37 ◦C by using a circulatory bath. The test compounds and controls were randomly applied to six 4 × 3 cm marked portions of nylon organdy strip, which was positioned over the six, membrane-covered wells. A six-celled K & D module

containing five 6–15-day-old females per cell was positioned over the six wells, trap doors were opened and mosquitoes allowed access for 3 minutes, after which they were collected back into the module. Mosquitoes were squashed and the presence of green dye (or not) in the gut was used as an indicator of feeding. A replicate consisted of six treatments: four samples, DEET, and ethanol as solvent control. Five replicates were conducted per day using new batches of mosquitoes in each replication.


**3.3 Effects of AV-6-93 and diflurone on Ca2+-induced MPT**

The effect of AV-6-93 and diflurone on Ca2+-induced MPT was studied in order to evaluate their capacity to protect mitochondria against MPT opening by measuring the decrease in ∆, the increase in oxygen consumption, and the Ca2+-induced release of mitochondrial Ca2+, which are typical phenomena that follow the induction of MPT. The amount of Ca2+ used to induce MPT was 100 nmol/mg protein.

spectrophotometrically at 530 nm. As blanks, we used control reactions performed in the absence of mitochondria and ADP/Fe2+. The amount of TBARs formed was calculated using a molar extinction coefficient of 1.56 x 105 mol-1 cm-1 and expressed as nmol TBARs/mg

The cytotoxicity data were calculated as a mean ± S.E. Statistical analysis was performed using Student's t-test or one-way analysis of variance (ANOVA), followed by a Bonferroni

The mitochondrial experiments were performed using three independent experiments with different mitochondrial preparations. The values are expressed as means ± S.E. Means were compared using one-way ANOVA for multiple comparisons, followed by Tukey's test.

In primary rat cortical neurones, AV-6-93 at concentrations of 1 and 10 µM decreased MPP+ induced cell death by 75% and 56%, respectively (Fig. 2A). Diflurone exerted the protective ability only at the highest tested concentration, 10 μM, and decreased the MPP+-induced cell death by 35% (Fig. 2B). Neither AV-6-93 nor diflurone, added without MPP+, changed cell

AV-6-93 and diflurone (up to 100 μM) were studied for their effects on mitochondrial bioenergetics by evaluating several mitochondrial respiratory chain parameters (state 2, state 3, state 4, FCCP-stimulated respiration, RCR, ADP/O ratio, ∆, and phosphorylation

The effects of AV-6-93 on glutamate/malate-supported respiratory rates (state 2, state 3, state 4 and FCCP-stimulated respiration), respiratory indices RCR and ADP/O of rat liver mitochondria were almost non-existent and insignificant at concentrations of up to 100 µM (Table 1), indicating that the compounds did not significantly affect mitochondrial

These results are demonstrated in Table 2, where AV-6-93 and diflurone, at concentrations of up to 100 µM, did not significantly affect either the ∆ induced by glutamate/malate-

As for glutamate/malate-supported respiration, the effects of AV-6-93 and diflurone on succinate-supported respiratory rates (state 2, state 3, state 4 and FCCP**)** and respiratory indices RCR and ADP/O of rat liver mitochondria were not significantly affected (results not shown), further supporting the finding that these compounds did not affect

The effect of AV-6-93 and diflurone on Ca2+-induced MPT was studied in order to evaluate their capacity to protect mitochondria against MPT opening by measuring the decrease in ∆, the increase in oxygen consumption, and the Ca2+-induced release of mitochondrial Ca2+, which are typical phenomena that follow the induction of MPT. The amount of Ca2+

**3.2 Effects of AV-6-93 and diflurone on rat liver mitochondrial bioenergetics**

protein (Buege & Aust, 1978).

**2.10 Statistical analysis**

multiple comparisons test.

**3. Results**

bioenergetics.

mitochondrial bioenergetics.

Statistical significance was set at *p*< 0.05.

**3.1 Protection against the cell death induced by MPP+**

viability at the highest tested concentrations (Fig. 2).

rate) using glutamate/malate as the respiratory substrate.

dependent respiration or the phosphorylation time.

used to induce MPT was 100 nmol/mg protein.

**3.3 Effects of AV-6-93 and diflurone on Ca2+-induced MPT**

Fig. 2. Influence of AV-6-93 (AV) and diflurone (D) on MPP+-induced cell death in primary rat cortical neurons (A and B, respectively). Cell death measured by Trypan blue method. Data are presented as a mean S.E. p < 0.001 vs control, t-test, \*\*\* p < 0.001 vs MPP+, one-way ANOVA followed by Bonferroni multiple comparison's test.

Targeting the Mitochondria

by Novel Adamantane-Containing 1,4-Dihydropyridine Compounds 265

Fig. 3*.* Effect of AV-6-93 on rat liver MPT induced by Ca2+.

with three different mitochondrial preparations.

Evaluation was performed by measuring succinate-supported transmembrane potential (∆) (A), oxygen consumption (B), and mitochondrial Ca2+ fluxes (C). Additions of 100 nmol calcium/mg protein (Ca2+) and 10 mM succinate (Suc); additions of AV-6-93 at the concentrations of 1, 10, and 100 M (1, 10, 100) are indicated. Assays in the absence of Ca2+ (-Ca2+); assays in the presence of Ca2+ plus CsA (0.75 nmol/mg protein (CsA); assays in the presence of Ca2+ plus CsA + 1M AV-6-93 (CsA+ 1). The traces are representative of assays

These effects were due to the entry of Ca2+ into the electronegative mitochondrial matrix (Fig. 3C), followed by the efflux of H+ for restoring the ∆. Incubation of mitochondria with

Table 1*.* Effects of AV-6-93 and diflurone on the respiratory parameters (state 2, state 3, state 4, FCCP-stimulated respiration) and respiratory indices (RCR and ADP/O ratio) of rat liver mitochondria using glutamate/malate as respiratory substrate.

The values, which are given in percentage of control (% of control), correspond to the mean ± S.E. of the respiratory parameters, evaluated in three different mitochondrial preparations, at the different indicated situations. Control values are expressed in nmol O2**.** mg-1 protein min-1: state 2 = 7.1 ± 0.7; state 3 = 39.4 ± 4.0; state 4 = 5.62 ± 0.8; FCCP-stimulated respiration = 57.14 ± 10.4.

Table 2. Effects of AV-6-93 and diflurone on glutamate/malate-dependent transmembrane potential (∆) and phosphorylation time of rat liver mitochondria.

The values correspond to the mean ± S.E. of the Δ and the phosphorylation time, evaluated in three different mitochondrial preparations, at the different indicated situations.

The results of the effect of AV-6-93 on MTP protection are depicted in Fig. 3. Under control conditions, the addition of 10 mM succinate to mitochondrial suspensions produced a ∆ of about -216 mV (negative inside mitochondria) (Fig. 3A), corresponding to respiratory state 4 (Fig. 3B). The addition of Ca2+ led to a rapid depolarisation (decrease of ∆), followed by a partial repolarisation (recover of ∆), the subsequent total depolarisation of mitochondria (Fig. 3A), and an increase in respiratory state 4 (Fig. 3B).

0.0 100.0 ± 0.0 100 ± 0.0 100.0 ± 0.0 100.0 ± 0.0 6.8 ± 1.3 3.01 ± 0.1 1.0 100.0 ± 0.0 113.4 ± 6.7 94.4 ± 5.6 100.2 ± 8.7 6.5 ± 1.5 2.9 ± 0.1 10.0 105.0 ± 18.9 104.0 ±7.3 119.4 ± 10.0 101.9 ± 7.3 6.4 ± 1.0 2.8 ± 0.4 100.0 127.2 ± 31.9 109.9 ± 6.6 145.8 ± 44.5 91.2 ± 8.5 6.3 ± 2.7 2.8 ± 0.1

0 100.0± 0.0 100.0± 0.0 100.0± 0.0 100.0± 0.0 6.8 ± 1.3 3.01 ± 0.1 100 107.3 ± 4.8 95.9 ± 2.9 104.4 ± 5.27 94.4 ± 4.03 6.4 ± 1.2 2.9 ± 0.09

Table 1*.* Effects of AV-6-93 and diflurone on the respiratory parameters (state 2, state 3, state 4, FCCP-stimulated respiration) and respiratory indices (RCR and ADP/O ratio) of rat liver

The values, which are given in percentage of control (% of control), correspond to the mean ± S.E. of the respiratory parameters, evaluated in three different mitochondrial preparations, at the different indicated situations. Control values are expressed in nmol O2**.** mg-1 protein min-1: state 2 = 7.1 ± 0.7; state 3 = 39.4 ± 4.0; state 4 = 5.62 ± 0.8; FCCP-stimulated respiration

**ADP**

0 -220.5 ± 5.1 21.7 ± 2.2 -216.8 ± 2.6 33.0 ± 3.0 1 -219.0 ± 4.0 21.3 ± 1.9 -217.0 ± 4.3 32.7 ± 2.9 10 -220.6 ± 4.1 23.5 ± 2.1 -218.7 ± 4.2 32.5 ± 4.5 100 -216.0 ± 5.6 23.4 ± 1.1 -212.5 ± 4.7 39.0 ± 1.7

0 -220.5 ± 5.1 21.7 ± 2.2 -216.8 ± 2.6 33.0 ± 3.0 100 -218.0 ± 1.7 21.0± 0.3 -216.0 ± 1.4 32.8 ± 2.3

Table 2. Effects of AV-6-93 and diflurone on glutamate/malate-dependent transmembrane

The values correspond to the mean ± S.E. of the Δ and the phosphorylation time, evaluated

The results of the effect of AV-6-93 on MTP protection are depicted in Fig. 3. Under control conditions, the addition of 10 mM succinate to mitochondrial suspensions produced a ∆ of about -216 mV (negative inside mitochondria) (Fig. 3A), corresponding to respiratory state 4 (Fig. 3B). The addition of Ca2+ led to a rapid depolarisation (decrease of ∆), followed by a partial repolarisation (recover of ∆), the subsequent total depolarisation of mitochondria

in three different mitochondrial preparations, at the different indicated situations.

mitochondria using glutamate/malate as respiratory substrate.

**energisation**

potential (∆) and phosphorylation time of rat liver mitochondria.

(Fig. 3A), and an increase in respiratory state 4 (Fig. 3B).

**RCR ADP/O State 2 State 3 State 4 State**

**FCCP**

**Δ (mV) Phosphorylation**

**time (s) Glu/Mal**

**depolarisation Repolarisation**

**Oxygen consumption (% of control)**

**Compounds (µM)**

**AV-6-93**

**Diflurone**

= 57.14 ± 10.4.

**Compounds (µM)**

**Diflurone**

**AV-6-93**

Fig. 3*.* Effect of AV-6-93 on rat liver MPT induced by Ca2+.

Evaluation was performed by measuring succinate-supported transmembrane potential (∆) (A), oxygen consumption (B), and mitochondrial Ca2+ fluxes (C). Additions of 100 nmol calcium/mg protein (Ca2+) and 10 mM succinate (Suc); additions of AV-6-93 at the concentrations of 1, 10, and 100 M (1, 10, 100) are indicated. Assays in the absence of Ca2+ (-Ca2+); assays in the presence of Ca2+ plus CsA (0.75 nmol/mg protein (CsA); assays in the presence of Ca2+ plus CsA + 1M AV-6-93 (CsA+ 1). The traces are representative of assays with three different mitochondrial preparations.

These effects were due to the entry of Ca2+ into the electronegative mitochondrial matrix (Fig. 3C), followed by the efflux of H+ for restoring the ∆. Incubation of mitochondria with

Targeting the Mitochondria

by Novel Adamantane-Containing 1,4-Dihydropyridine Compounds 267

Fig. 4. Effect of AV-6-93 on membrane lipid peroxidation of rat liver mitochondria induced

**0 10.2 ± 1.0 1 9.6 ± 1.0 10 8.5 ± 0.5 20 7.6 ± 1.0 50 5.2 ± 2.1 100 3.1± 1.9\***

Table 3. Effect of AV-6-93 on membrane lipid peroxidation of rat liver mitochondria induced by the pro-oxidant pair ADP/Fe2+ evaluated by TBARs assay. The data correspond to the

Studies examining the importance of mitochondrial pathophysiology in neurodegeneration provide a target for additional treatments with agents that improve mitochondrial function, protect MPT, and/or exert antioxidant activity (Petrozzi et al., 2007). These studies lead to novel approaches in the treatment of neurodegenerative diseases, such as Parkinson's

The aim of the present study was to examine the abilities of two novel adamantanecontaining DHP analogues, AV-6-93 and diflurone, to protect against cell death induced by mitochondrial toxin MPP+ and beneficially influence mitochondrial processes in an attempt

First, we examined how both compounds acted in primary cortical cultures in response to MPP+. AV-6-93, at concentrations of 1 and 10 µM, significantly protected against MPP+ induced cell death by 75% and 56%, respectively, whereas diflurone protected against cell death by 35% at a concentration of 10 µM. Neither AV-6-93 nor diflurone, added without

**TBARs (nmol/mg protein/10 min)**

by the pro-oxidant pair ADP/Fe2+ evaluated by oxygen consumption.

**AV-6-93 (M)**

mean ± S.E. of three independent experiments. \**p*< 0.05 vs control (in the absence of AV-6-93).

disease, with disease-modifying drugs.

MPP+, changed cell viability.

to identify putative antiparkinsonian drugs.

**4. Discussion**

AV-6-93 concentrations of up to 100 µM for 3 min before energisation with succinate prevented total depolarisation of mitochondria (Fig. 3A), the increase in respiratory state 4 (Fig. 3B), and the release of mitochondrial Ca2+ (Fig. 3C), suggesting that this compound has a high ability to protect mitochondria against MPT induction. Incubation of mitochondria with 0.75 nmol/mg protein, CsA, a specific inhibitor of MPT (Broekemeier et al., 1989), for 2 min before energising with succinate, either in the absence or presence of 1 μM AV-6-93, completely blocked mitochondrial depolarisation (Fig. 3A), the increase in respiratory state 4 (Fig. 3B), and the Ca2+-induced release of mitochondrial Ca2+ (Fig. 3C). These data show that these effects had been induced by MPT. In contrast to AV-6-93, diflurone, in the same concentration range, did not prevent either the depolarisation of mitochondria or the release of mitochondrial Ca2+ (results not shown), indicating that this compound did not protect mitochondria against MPT.

#### **3.4 Effects of AV-6-93 and diflurone on mitochondrial oxidative stress**

The effects of AV-6-93 and diflurone on mitochondrial oxidative damage were assessed by detecting the mitochondrial membrane lipid peroxidation induced by the pro-oxidant pair ADP/Fe2+. Lipid peroxidation was evaluated by measuring oxygen consumption (Fig. 4) and TBARs formation (Table 3). In the absence of AV-6-93 and after the addition of the prooxidant pair, it is possible to distinguish two-phase kinetics in oxygen consumption: an initial lag phase, characterized by slow oxygen consumption lasting about 2 min, is followed by a rapid oxygen consumption phase. The lag phase is probably related with the time required for the generation of a sufficient amount of the perferryl ion complex (ADP-Fe2+- O2 ADP-Fe3+-O2-), which has been suggested to be responsible for the initiation of lipid peroxidation. The rapid oxygen consumption phase is probably due to the oxidation of the polyunsaturated fatty acid acyl chain of membrane phospholipids by reactive oxygen species (ROS) and, consequently, due to the propagation phase of lipid peroxidation (Sassa et al., 1990). AV-6-93 concentrations up to 100 M enlarged the lag phase of slow oxygen consumption before the oxygen uptake burst induced by the ADP/Fe2+ complex and increased the rate of the rapid oxygen consumption phase (Fig. 4), suggesting that the compounds affected both the initiation and the propagation of lipid peroxidation of mitochondrial membranes.

These results agree with the quantitative evaluation of TBARs formation performed to confirm the protective effects of AV-6-93. The data in Table 3 show that the kinetics of TBARs formation induced by ADP/Fe2+ are similar to that observed for oxygen consumption. The same range of AV-6-93 concentrations used in the oxygen consumption assays also affected TBARs formation. TBARs formation in the absence of ADP/Fe2+ was negligible (0.44 ± 0.25 nmol/mg of protein). In contrast to AV-6-93, diflurone, in the same concentration range, did not affect oxygen consumption induced by the ADP/Fe2+ complex or TBARs formation (results not shown), indicating that this compound has no capacity to protect mitochondria against the lipid peroxidation induced by the pro-oxidant pair ADP/Fe2+.

Lipid peroxidation was evaluated by oxygen consumption and initiated by adding 1 mM ADP/0.1 mM Fe2+ to mitochondrial suspensions (Fig. 4). The traces represent typical direct oxygen consumption recordings of three experiments obtained from different mitochondrial preparations; controls in the absence of ADP/Fe2+ (–ADP/Fe2+); assays in the presence of AV-6-93 at the concentrations 1, 10, 20, 50, 100 μM (1, 10, 20, 50, 100).

Fig. 4. Effect of AV-6-93 on membrane lipid peroxidation of rat liver mitochondria induced by the pro-oxidant pair ADP/Fe2+ evaluated by oxygen consumption.

Table 3. Effect of AV-6-93 on membrane lipid peroxidation of rat liver mitochondria induced by the pro-oxidant pair ADP/Fe2+ evaluated by TBARs assay. The data correspond to the mean ± S.E. of three independent experiments.

\**p*< 0.05 vs control (in the absence of AV-6-93).


**Abstract**

The problem of migration into the Republic of South Africa took shape at the dawn of democracy and it has since then become overly complex to manage. South Africa has witnessed an unprecedented, undocumented population over the past three decades. The illegal migrants are economic migrants who enter the republic illegally for greener pastures. There are possible repeat deportees commonly known as the "revolving door" syndrome. The number of repeat deportees and the reasons for their continuous return is not known. The deportation process is currently faced with numerous challenges of re-entry after deportation and serious budget cuts to fulfil the mandate. This has significant impacts on the management of deportation in the country. This chapter provides various aspects of immigration and deportation from a South African context and also identifies challenges faced to manage the deportation of illegal immigrants.

**Keywords:** asylum seekers, corruption, fraudulent entry, social networks

### **1. Introduction**

There is only a small percentage of the world's population that live outside their countries of birth or origin, but migration is still an important phenomenon on the political agenda of some countries. The movement of people across borders is a potential problem for almost all countries as it is nearly impossible to completely control immigration flows. The management of deportation is a mandatory function of the Department of Home Affairs (DHA) in the Republic of South Africa in terms of Section 34 of the Immigration Act 13 of 2002 as amended [1]. The DHA has two core businesses which are civic services and Immigration Services (IMS). Deportation management falls under the branch of IMS. IMS has further three business units namely, Port Control, Permitting (Visas), Asylum Seekers Management and Inspectorate. Inspectorate is established in terms of Section 33 of the Immigration Act 13 of 2002 as amended. The management of deportation is one of the functions of the Inspectorate, which is the law enforcement arm of the department [2]. The responsibility of the inspectorate is to comply with the Immigration Act. It is the responsibility of the deportation directorate to deport Illegal Foreigners (IFs) who are detected and arrested for contravening the Immigration Act.


**Abstract**

In monitoring and supervision schemes, fault detection and diagnosis characterize high efficiency and quality production systems. To achieve such properties, these structures are based on techniques that allow detection and diagnosis of failures in real time. Detection signals faults and diagnostics provide the root cause and location. Fault detection is based on signal and process mathematical models, while fault diagnosis is focused on systems theory and process modeling. Monitoring and supervision complement each other in fault management, thus enabling normal and continuous operation. Its application avoids stopping productive processes by early detection of failures and by applying real-time actions to eliminate them, such as predictive and proactive maintenance based on process conditions. The integration of all these methodologies enables intelligent monitoring and supervision systems, enabling real-time fault detection and diagnosis. Their high performance is associated with statistical decision-making techniques, expert systems, artificial neural networks, fuzzy logic and computational procedures, making them efficient and fully autonomous in making decisions in the real-time operation of a production system.

**Keywords:** automatic control, availability, intelligent systems, monitoring, predictive maintenance, supervision


*4.3. Molecular Dynamics Simulations*

The CHARMM 36 force field [34] was used for the simulations summarized in Table 1 using the GROMACS software package version 5.1.3 [35]. The YqhD dimer bound to Zn2+ and NADP/H cofactors was centered in a cubic periodic box (~11 nm3 ) and set to have a distance larger than 1 nm from any side of the box. Solvent molecules having any atom within 0.15 nm from the protein were removed. TIP3P model [36,37] was used for explicit water solvent. Sodium counter ions were added by replacing the solvent molecules at the sites of most negative electrostatic potential to provide the box with a total charge of zero. The protonation state of residues was assumed to be the same as that of the isolated amino acids in solution at pH 7. The LINCS [38] algorithm was used to constrain all bond lengths and the SETTLE [39] algorithm was used for the water molecules. Electrostatic interactions were calculated using the Particle Mesh Ewald method [40]. For the calculation

of long-range interactions, a grid spacing of 0.12 nm combined with a fourth-order B-spline interpolation was used to compute the potential and forces between grid points. A nonbonded pair-list cutoff of 1.4 nm was used and updated at every five time-steps. V-rescale thermostat [41] was used to keep the temperature at 300 K through a weak coupling of the system to an external thermal bath with a relaxation time constant τ = 0.1 ps. The pressure of the system was kept at 1 bar using Berendsen's barostat [42] with a time constant of 1 ps. A time step of 2 fs was used to integrate the equations of motion.

**Table 1.** Simulation summary for the YqhD homodimer in aqueous solution.

First, the simulated systems were energy minimized, using the steepest descent algorithm, for at least 5000 steps to remove clashes between atoms that were too close. After energy minimization, all atoms were given an initial velocity obtained from a Maxwell-Boltzmann velocity distribution at 300 K to start the MD simulations. The system was initially equilibrated by 30 ps with position restraints on the heavy atoms of the dimer to allow relaxation of the solvent molecules. After the equilibration procedure, position restraints were removed, and the system was gradually heated from 50 K to 300 K during 200 ps of simulation. The equilibrated structure was used to perform MD simulations in the NPT ensemble for 20 ns at 300 K. The final conformation of the 20 ns isothermalisobaric NPT ensemble was used to set up five sets of 200 ns independent simulations in the canonical NVT ensemble initiating with five different velocities.

The starting coordinates were adopted from the crystal structure which has modified cofactor (NADPH(OH)2); hence, the equilibrated conformations of YqhD bound to NADP and NADPH cofactors were obtained after a 20 ns NPT simulation, which was used as the starting point for the final production run of 200 ns long NVT simulations. The starting crystallographic coordinates of the YqhD homodimer were used as the reference structure for the analysis of the trajectories to characterize the conformational changes induced by a change in the oxidation state of NADP/H cofactor. The analysis was focused on three sets/sub-structures within the simulation data, considering dimer, monomer, and domain in a 1 µs aggregated trajectory for each cofactor oxidation state (NADP and NADPH). Structural analysis includes the root mean square deviation (RMSD), root mean square fluctuation (RMSF), radius of gyration (Rg), and cofactor binding interactions (Zn+2 and NADP/H), comparing these with respect to the crystal structure.


**1. Introduction**

According to the 2020 biotechnology industry white paper, the global pharmaceutical market in 2019 reached approximately US \$1.25 trillion, an increase of 4% compared to 2018. It takes an average of 10–15 years and the US \$300–800 million to develop a new drug. Under increasingly strict global supervision, drug development has become more complicated, requiring numerous resources and equipment support. Therefore, reducing development and time costs, and ensuring compliance with strict regulations and standards at the same time, has become a common problem faced by global biotechnology and pharmaceutical companies.

In the entire drug development process, the clinical trial phase requires one-half to two-thirds of the development cost and time. Due to this, clinical trials have been outsourced to the Contract Research Organization (CRO) in recent years. According to Frost & Sullivan estimates, the global CRO turnover in 2019 reached

approximately US \$49.74 billion, an increase of 8.5% compared to 2018. It can be seen that the clinical trial service industry is very important for biotechnology and medicine. Furthermore, the CRO industry is a knowledge-intensive industry. Professional knowledge and skills are its main requirements, which are also key to the competitiveness of enterprises. Additionally, the accumulation of experience and knowledge, and the maintenance of external relations are both important assets for an enterprise in the clinical trial industry. Therefore, improving the competitiveness of an organization through effective knowledge management has become one of the most important issues in business management.

Previous research on CRO mainly focused on its strategy, business, and operations model, and seldom discussed the knowledge management of CRO. Therefore, this research will focus on the CRO industry and select companies that provide clinical trial services in Taiwan to study their knowledge management practices and the impact of influencing factors on knowledge management activities.

The issues to be discussed in this research are as follows:

In the following section, an overview of the CRO industry is introduced, as well as related literature on influential factors of knowledge management and knowledge management activities. The research framework is then introduced in the third section based on the research purpose and the literature review results. Besides, in this section, the research method, research objects, and data collection methods of this study are further explained. The fourth section describes the research cases. In the fifth section, seven research propositions are proposed and are followed by discussions. Finally, conclusions are drawn and the theoretical implications are depicted in the last section.


**5. The place for the analyst**

*"In choosing theater as a metaphor, psychic reality was perhaps following in the footsteps of Anna O., who, at the turn of the century, evoked her "free associations", on the occasion of her treatment with Breuer, as her "private theater". For the analyst, it is a theater that his patients want to share with him and which he is invited to play in various roles. But, unlike Breuer, who was frightened by the role Anna O. made him play, the analyst tries to observe his own inner theater well and interpret it as best he can before playing his patient's." [57]*

The term created by those analysts, who want to validate the social speech instead of analytical work, is mistaken by definition when the gender affirmative is the confirmation of gender; however, it is not possible to confirm what is neutral, out of binary, or yet, out of innate human bissexuality defined by Freud in the perverse polymorph sexuality [2]. And put those searching for a work inside the psychoanalytic approach in a delicate position of gender critical, as if thinking about gender was something critical, and maybe it is, critical in a sense, referring to a crisis. It is critical to think about the denying of the body in favor of a psychotic society, that is, the one that has seen the difference of others as an enemy, a persecutor that must be eliminated.

Therefore, the present authors disagree with that term created for those psychoanalysts who value the analytical work, "where there was id, that have an Ego," that is, the reality principle prevails over the pleasure principle [31].

The "trans" children in this sense do not transit between natural genders, as they are supposed to do in childhood [31]. Given that according to gender affirmative they


*Article* **A Hybrid Asynchronous Brain–Computer Interface Based on SSVEP and Eye-Tracking for Threatening Pedestrian Identification in Driving**

**Jianxiang Sun and Yadong Liu \***

College of Intelligence Science and Technology, National University of Defense Technology, Changsha 410073, China

**\*** Correspondence<EMAIL_ADDRESS>
**Abstract:** A brain–computer interface (BCI) based on steady-state visual evoked potential (SSVEP) has achieved remarkable performance in the field of automatic driving. Prolonged SSVEP stimuli can cause driver fatigue and reduce the efficiency of interaction. In this paper, a multi-modal hybrid asynchronous BCI system combining eye-tracking and EEG signals is proposed for dynamic threatening pedestrian identification in driving. Stimuli arrows of different frequencies and directions are randomly superimposed on pedestrian targets. Subjects scan the stimuli according to the direction of arrows until the threatening pedestrian is selected. The thresholds determined by offline experiments are used to distinguish between working and idle states of the asynchronous online experiments. Subjects need to judge and select potentially threatening pedestrians in online experiments according to their own subjective experience. The three proposed decisions filter out the results with low confidence and effectively improve the selection accuracy of hybrid BCI. The experimental results of six subjects show that the proposed hybrid asynchronous BCI system achieves better performance compared with a single SSVEP-BCI, with an average selection time of 1.33 s, an average selection accuracy of 95.83%, and an average information transfer rate (ITR) of 67.50 bits/min. These results indicate that our hybrid asynchronous BCI has great application potential in dynamic threatening pedestrian identification in driving.

**Keywords:** brain–computer interface (BCI); steady-state visual evoked potential (SSVEP); electroencephalography (EEG); threatening pedestrians; eye-tracking


Cumin (*Cuminium cyminium* L.): A Seed Spice Crop with Adopted Production Technology in Cumin Cultivated Regions

*Navjot Singh Brar, Prakash Mahala, Kartik Sharma, Parmdeep Singh Dhanda, Alpa Yadav, Meenakshi Sharma and Prashant Kaushik*


**2. Theory**

32 Photonic Crystals – Innovative Systems, Lasers and Waveguides

During recent years different approaches have been done by scientists for generating of slow light. Electromagnetically induced transparency (EIT) is the first technique for achieving group delay and slow light in vapor mediums [5-9]. The group delay of incident pulses in an EIT system was studied first by Kasapi *etal* in lead vapor. Also, in 1999 Hau

As a reason of complicated situation for slow light generation in EIT systems, it is important for real applications that these phenomena could be occurred in room temperature and in

Therefore, Spectral holes due to coherence population oscillation (CPO) in room temperature and in solids, can be considered as one of the interesting methods for slow light generation [11]. Slow light generation based on CPO has been studied in various material systems. So, ultraslow light, 57.5m/s, in ruby crystal has been studied by Boyd's group at

On the other hand, during recent years a new kind of optical waveguides have been considerable attracted both theoretical and experimental attention due to their intense applications not only in data transferring but also in optical data processing [13]. Coupled cavity optical waveguides (CCWs) can be considered as the latest proposal mechanism for optical waveguiding which have been introduced by Yariv *etal* at Caltech [14]. This new kind of waveguides is based on the periodic dielectric structures as photonic crystals, which is separated by the high quality factor cavities that are coupled to each of the nearest neighbor in multiple spatial dimensions [15]. Due to existence of the cavities along the structure and as a result of the overlapping of the evanescent fields, light can be propagated through the CCWs [16]. Actually, it was investigated theoretically and experimentally that CCWs exhibit more advantages over conventional optical waveguides. One of the most important features of CCW is that due to strong optical confinement in defect medium, and high slope of transmission at resonance wavelengths, group velocity at the edge of each resonance modes can be reduced considerably [17]. Slow light generation in CCWs offers several practical advantageous like, design freedom, direct integration with other optoelectronics devices and ability to slow down light in a desired region of wavelengths at

On the other hand, it should be noticed that the CCWs structures for stopping light suffer a fundamental trade off between the transmission and the optical delay bandwidth [19,20]. Therefore, in the field of slow light technology, delay-bandwidth product is an important

Furthermore, from system points of view, in future photonic circuits, adjustability of the optical properties of components is a great and important bottleneck which many scientists have been proposed special methods. In CCWs, the optical properties such as group velocity, dispersion and its higher order can be modulated through different mechanism such as electro optic effect, free-carrier injection and thermo optic effect [21]. As a consequence of necessity for all optical networks in future, dynamic all optical processing, controlling light by light, can be considered as one of the crucial bottlenecks for future all

s delay in an EIT system which consists of condensed cloud of sodium

*etal* showed 7.05

Rochester [12].

room temperature [18].

optical systems [22].

parameter which should be considered greatly.

atoms [10].

compact and solid materials.

1D-NCCW is formed by placing optical resonator in a linear array, to guide light through whole of the structure by photon hopping between adjacent resonators. As a result of overlapping the evanescent field in the defect medium, cavity zone, electric field enhancement in this region can be obtained. Tight binding (TB) approximation (like using it in solid state physics) is used to describe the mechanism of waveguiding of the structure.

The basic structure for 1D-NCCW is as:


**1. Introduction**

Concrete is the most widely used building material due to its strength, adaptability, low maintenance requirements during the lifetime of structures, and the economic and extended accessibility of its components [1], which makes it very difficult to replace in many infrastructure applications [2]. In fact, globally, concrete production is estimated at approximately 25 billion tonnes per year [3].

However, a relevant challenge is the amount of harmful emissions produced by concrete [4]. Actually, cement manufacturing and concrete production account for 6–9% of global man-made CO2 [5, 6]. Furthermore, by 2050, this value is projected to increase by 4% due to an increase of 12–23% in cement consumption [7]. Cement production generates, as well, high NOx and SOx emissions, which contribute to the development of acid rain, deterioration of public health and global climate change [8, 9].

Additionally, the high demand for natural aggregates used in the concrete mix generates a loss of vegetation and fauna, a decrease in air quality due to emissions of particulate matter, loss of fertile soil, risks of contamination of groundwater and deterioration of the life quality of people living near the extraction sites [10]. Furthermore, the extraction of aggregates can cause morphological alterations in the shape of the channel, bottom and banks, which has repercussions upstream through rebound erosion, generating a considerable increase in the velocity and shear stress of the flow and favouring the erosion processes in the riverbed [11].

The negative impacts related with the concrete material can be minimised by adopting a sustainable approach [12], where sustainability is understood as meeting the needs of the present without compromising the ability of future generations to meet their own needs [13]. In this context, sustainable engineering implies not only the use of sustainable materials, but a sustainable engineering system, where different engineering stages and processes can include factors such as reduction of environmental impacts, economic accessibility and access to the engineering solution regardless of the geographical location [14]. Therefore, from this holistic approach, the result is an alternative with ae technical, environmental, economic and social balance [15].

As important as it is, sustainability related with the concrete material is more than reducing the amount of cement in concrete mixes. Actually, sustainable characteristics have been reported by optimising the design of structures [16–18], replacing natural aggregate with different types of waste such as recycled crushed concrete, marble waste [19] or waste foundry sand [20] and incorporating nontraditional materials such as fibres [21, 22], biological material [23], hazardous waste material [24] or nanomaterials into concrete [25].

The objective of this book chapter is to analyse different types of contributions to a sustainable development using the concrete material. Although some examples are mentioned for the case of reducing the amount of cement directly in the concrete mixes, the analysis is focussed on the other approaches presented in Section 3 and analysed in Section 4 using different examples, most of them proposed from the concrete laboratory of the University of Concepción, Chile. Related with this, Section 2 presents the fundamental concept of geo-dependency, which is directly related with the development of practical and useful sustainable concrete alternatives.


**Author details**

Kimberly A. Heinemann1 \*, Heather Bolanos2 and Jennifer S. Griffin3

\*Address all correspondence to<EMAIL_ADDRESS>
1 Hunter College of the City University of New York, CUNY, New York, NY, United States of America

2 Reach for the Stars Learning Center, Brooklyn, NY, United States of America

3 Patoka Community Unit School District, Patoka, IL, United States of America


Preface

Recent advances in the field of plant biotechnology succeeded in altering various metabolic pathways via gene and metabolic engineering approaches to introduce novel compound production in plants. Molecular biotechnology strategies can be used to improve plant biomass by enhancing cellulose and starch for biofuel production. Biotechnology and genetic engineering approaches are useful for the improvement of crops for biotic and abiotic stress tolerance and increased food production. As such, this book provides updates on recent developments in plant science highlighting major trends in botany, particularly in plant biotechnology, alternative energy, food industry, environment, and medicine. *Botany - Recent Advances and Applications* presents comprehensive information about current advances in plant science. It contains five chapters by eminent research scholars that cover a vast range of research on soil organic carbon sequestration of tropical grasses and their potential use as biomass energy crops to mitigate global environmental changes. The book also highlights nonedible seeds as a potential renewable source of biodiesel production to control the global energy crisis. Finally, the book discusses the importance of microRNA as a potential tool for improving agronomic traits of several economically important crop plants as well as examines the uses, methods, and advantages of recombinant DNA technology and plant biotechnology for the development of plant-based vaccines.

We are enormously grateful to our publisher IntechOpen and the authors who contributed their work to this book. We also extend our sincerest appreciation to Editor and Author Service Manager Sara Gojevic-Zrnic, who was instrumental in the preparation and publication of this volume.

> **Dr. Bimal Kumar Ghimire** Department of Crop Science, College of Sanghuh Life Science, Konkuk University, Seoul, Korea