**Table 6.** Overview of anticonvulsant effects of ghrelin-R ligands in experimental epilepsy models. D1R: Dopamine 1 receptor; i.v.: intravenous; i.h.: intrahippocampal; IHKA: intrahippocampal kainic acid; i.p.: intraperitoneal; KA: kainic acid; min: minute; pilo: pilocarpine; Ref: reference; SP: [D-Arg1,D-Phe5,D-Trp7,9,Leu11]substance P.

**Table 6.** *Cont.*

#### **5. Molecular Mechanisms-of-Action**

*5.1. Mechanisms of Ghrelin's Anticonvulsant Action*

Ghrelin-R expression is dynamic and may be influenced by the presence of a disease state, or may depend on exposure to ghrelin [54,55,134], which are both relevant in the context of ghrelin administration in seizure and epilepsy models. Neither ghrelin nor pilocarpine altered hippocampal ghrelin-R mRNA expression in the pilocarpine rat model [28], while another group showed a decrease in the hippocampal ghrelin-R mRNA expression in pilocarpine-treated rats, which was restored upon ghrelin administration [30].

Hippocampal Akt signaling was decreased in a pilocarpine rat model, which could be restored by ghrelin administration [28,30]. Akt is a downstream target of the ghrelin-R, which can be activated both by Gαq signaling and β-arrestin recruitment. The ghrelin-R antagonist, EP-80317, restored the increased hippocampal phosphorylation levels of the other canonical downstream target ERK in the 6-Hz mouse model [131].

Up until now, the exact signaling pathways responsible for anticonvulsant effects downstream of ghrelin-R have remained elusive, but a few possibilities exist, on the basis of previous findings. Not only ghrelin-R agonists, but also ghrelin-R inverse agonists exerted anticonvulsant effects, and ghrelin-R KO mice were protected from seizures [116,124]. The truncated ghrelin variant, ghrelin (1–5) amide, shows similar EC50 values compared to ghrelin with regard to the ghrelin-R signaling pathways, but is unable to internalize the ghrelin receptor, and was not able to exert anticonvulsant effects [116]. Because of these intuitively irreconcilable observations, a novel concept emerged, hypothesizing that the absence of the ghrelin-R on the cell surface was responsible for exerting ghrelin's anticonvulsant effect [116]. We showed that a Gαq and Gα12 selective biased ligand of the ghrelin-R, YIL781, increased seizure severity in a kindling model [125]. Given these observations, β-arrestin recruitment remains the most probable pathway involved in ghrelin-R-mediated anticonvulsive effects and it requires further investigation.

However, we cannot completely exclude that G-protein-dependent signaling may be required for ghrelin-R-mediated anticonvulsant effects, as Akt and ERK activation have been described [30,32,135]. However, this would not fit with the notions of ghrelin-R KO mice being protected from seizures, and inverse agonists exerting anticonvulsant effects; in these cases, there is no G-protein-dependent signaling downstream of the ghrelin-R. Finally, one could hypothesize that the possibility exists that the signaling pathways downstream of β-arrestin may be responsible for ghrelin's anticonvulsant effects. Indeed, Akt and ERK are mediators that can also be activated via β-arrestin-dependent signaling. Nonetheless, and also here, the data obtained from the experiments with ghrelin-R KO mice and inverse agonists [116] suggest otherwise, and point towards an absence of signaling, which is imperative for ghrelin's anticonvulsant effects.

#### *5.2. Mechanisms of Neuroprotection*

Ghrelin increased the number of surviving neurons in the CA1 and CA3 hippocampal regions in the pilocarpine rat model [28]. Pilocarpine reduced the apoptotic repressor, B-cell lymphoma 2 (Bcl-2), increased the proapoptotic member Bcl-2-associated X protein (Bax), and increased cleaved caspase-3, crucial in apoptosis. Ghrelin was able to restore these markers, and may thus exert neuroprotection through antiapoptotic effects [28].

This latter finding was confirmed in the i.p. KA mouse model, in which ghrelin decreased cleaved caspase-3 immunoreactivity in pyramidal CA1 and CA3 neurons, and restored neuronal loss in CA1 and CA3 [29]. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive cells were abundantly present in vehicle-treated KA mice, but no TUNEL-positive cells could be observed in ghrelin-treated KA mice. All of the above described effects were dependent on ghrelin-R, as they were reversed by the concurrent administration of a ghrelin-R antagonist [29]. A study conducted by Zhang and colleagues confirmed the necessity of ghrelin-R availability in order for ghrelin to exert its neuroprotective effects [30]. Ghrelin significantly rescued neuronal cell loss in CA3, and inhibited cleaved caspase-3 activation, mediated via the phosphorylation of Akt [30].

A two-week-long administration of the ghrelin-R agonist, macimorelin, in the IHKA mouse model, exerted anticonvulsant effects on spontaneous recurrent seizures, but did not increase neuronal survival in the CA1, CA3, and DG of the hippocampus. This could be due to the omission of pretreatment, as the onset of the treatment commenced 24 h after SE induction, or due to the additional two-week wash-out in this study [126]. A lower dose of macimorelin, administered prior to SE, was found to increase neuronal survival and decrease apoptosis in the DG in the pilocarpine rat model, but not in CA1 [31]. Additionally, this study was able to demonstrate neuroprotective effects exerted by macimorelin, but not anticonvulsant effects against the pilocarpine-induced SE.

The possibility should be considered that ghrelin exerts neuroprotective effects via the activation of a variety of signaling pathways mediated through the employment of Gproteins, whereas the rapid and subsequent internalization via β-arrestin signaling may be responsible for anticonvulsant effects. Unraveling which ghrelin-R downstream signaling pathway is responsible for a particular effect may be obtained by doing further experiments in genetic models, or by using biased ligands that selectively activate a subset of pathways while leaving others untouched.

#### *5.3. Inflammation*

Inflammation is a major hallmark of epileptogenesis, seizures, and chronic epilepsy. This ranges from infiltration of the inflammatory cells and the release of proinflammatory mediators to widespread gliosis [134–137]. Given the fact that inflammation is known to progress the development of epilepsy [138], one of the presumed mechanisms-of-action of ghrelin may rely on its ability to attenuate inflammation, stemming from both direct central actions, and through peripheral anti-inflammatory effects [139].

Ghrelin significantly reduced the elevated plasma calcitonin gene-related peptide (CGRP), substance P, interleukin (IL)-6, tumor necrosis factor (TNF)-α, and IL-1β in the PTZ rat model [117,118]. Additionally, ghrelin inhibited KA-induced increases in TNF-α, IL-1β and cyclooxygenase-2 (COX2) mRNA levels in CA1 and CA3 in the i.p. KA mouse model, mediated via the ghrelin-R [29]. Ghrelin restored KA-induced increased matrix metalloproteinase 3 levels, which is an important mediator of inflammation and neuronal cell death [29]. Additionally, KA-induced increases in microglia and glial fibrillary acidic protein (GFAP) immunoreactivity in CA1 and CA3 three days after SE were inhibited by ghrelin [29]. This was not detectable after a two-week wash-out following macimorelin administration in the IHKA mouse model, in which macimorelin was administered after KA, and not as a pretreatment [126]. Another study showed that ghrelin administration decreased cortical TNF- α and NF-κB expression in the pilocarpine rat model [140]. However, ghrelin did not alter serum levels of galanin, fibroblast growth factor (FGF-2), IL-6, TNF-α, and IL-1β in the Wag/Rij rat model with nonconvulsive absence seizures [114].

#### *5.4. Oxidative Stress*

Seizures induce oxidative stress, which, in turn, exacerbates seizures (reviewed by [141]). Ghrelin prevented the PTZ-induced decrease in the catalase activity in both the CNS and erythrocytes, and prevented the augmentation in thiobarbituric acid reactive substances levels, a measure for lipid peroxidation [142]. Additionally, ghrelin normalized superoxide dismutase levels, an enzyme responsible for clearing superoxide anion in the erythrocytes, brain, and liver [142]. These data suggest that ghrelin protects against oxidative stress caused by PTZ. It remains unclear whether the effects of ghrelin on decreasing oxidative stress are caused directly, or because ghrelin is anticonvulsant, and the lower number of seizures leads to decreased oxidative stress. However, in the WAG/Rij rat model with nonconvulsive absence seizures, ghrelin was not anticonvulsant, but it still reduced the malondialdehyde [114].

#### **6. Functional Implications of Diminished Ghrelin-R Signaling in the Context of Excitability**

While several molecular mechanisms-of-action have been described, it remains unknown how these contribute to the anticonvulsant effects of ghrelin, and how they lead to an overall decrease in the brain excitability. The heteromerization of the ghrelin-R with other receptors can lead to the preferential recruitment of other noncanonical signaling pathways [143]. Ghrelin, as well as other signaling molecules, may exploit this phenomenon for inducing ghrelin-R-mediated anticonvulsant effects.

Ghrelin-R activation results in intracellular Ca2+ increases through the canonical Gαq protein [59,60]. One possible mechanism-of-action would be a decrease in intracellular Ca2+ in ghrelin-R-expressing neurons. Elevated levels of intracellular Ca2+ are associated with epileptiform activity and epileptogenesis [144,145]. Therefore, a reduction in intracellular Ca2+ may be an interesting putative mechanism for seizure suppression in the absence of ghrelin-R signaling. Various studies have shown the differential effects of ghrelin on neuronal excitability and synaptic transmission, which all support that ghrelin acts in a brain-region-specific manner [146–148].

The ghrelin-R is expressed in both excitatory neurons, as well as in inhibitory interneurons in the dorsal CA1. It was recently shown that a selective increased expression of the ghrelin-R in excitatory neurons was detrimental for learning and memory in mice, while an increased expression of the ghrelin-R in interneurons had a beneficial effect [149]. It remains to be uncovered if a dual effect on excitability also exists by, for instance, decreasing Gαq signaling in excitatory neurons via β-arrestin, while increasing Gαq signaling in inhibitory interneurons. Indeed, while GPCRs may be associated with several signaling pathways, these signaling pathways are not always all operative in the same cell. Thus far, the knowledge concerning the cell-specific expression of signaling pathways downstream of the ghrelin-R is lacking and requires further studies.

#### **7. Conclusions and Future Perspectives**

Ghrelin is increasingly recognized as a potential important player in seizures and epilepsy. Most studies show lower ghrelin levels in patients suffering from epilepsy, or lower ghrelin levels after a seizure. The exact implications of plasma ghrelin level alterations in epilepsy have remained, up until now, unknown, and should be further investigated in light of its treatments as well, including the KD. It is increasingly evident that there may be important differences between AG and DAG. This may advocate for future investigations of both isoforms of ghrelin in epilepsy, or for studying further whether the contributions of GOAT expression and the local reacylation of DAG are relevant for seizure control.

With only a few exceptions, ghrelin and synthetic agonists of the ghrelin-R are anticonvulsant in seizure, epilepsy, and SE models. The notion that both agonists and inverse agonists were anticonvulsant stirred up the discussion concerning the signaling pathways responsible for ghrelin-R-mediated anticonvulsant effects. The hypothesis that β-arrestin recruitment is involved should be more thoroughly investigated to confirm the relevance of this pathway. Overall, the complexity of ghrelin-R signaling, and the extensive list of other factors possibly influencing it, highlight the need for further investigations into the mechanism behind ghrelin-induced anticonvulsant effects.

**Author Contributions:** Conceptualization, A.B., D.D.B., R.K. and I.S.; Investigation, A.B.; Resources, D.D.B., R.K. and I.S.; Writing—Original Draft Preparation, A.B.; Writing—Review & Editing, A.B., D.D.B., R.K. and I.S.; Visualization, A.B.; Supervision, D.D.B., R.K. and I.S.; Project Administration, A.B.; Funding Acquisition, A.B., D.D.B., R.K. and I.S. All authors have read and agreed to the published version of the manuscript.

**Funding:** This research was supported by the scientific Willy Gepts fund of the UZ Brussel, the Queen Elizabeth Medical Foundation (ING prize), and the strategic research program of the Vrije Universiteit Brussel (SRP49). A Buckinx is a research fellow of the Fund for Scientific Research, Flanders (SB-FWO grant no. 1S84218N).

**Conflicts of Interest:** The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

#### **References**

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Section 2 Surgery in Clefts

**35**

**Chapter 3**

**Abstract**

**1. Introduction**

Surgery

Optimizing Outcomes in Cleft

*Eugene Park, Gaurav Deshpande, Bjorn Schonmeyr,*

Children born with cleft lip and palate in low- and middle-income countries (LMICs) have limited access to surgical care and suffer substantial morbidity as a result of delayed treatment. Charitable organizations have sought to tackle this problem through high-volume surgical missions, but these efforts have been fraught with high rates of complications and variable esthetic results. Over the past decade, Operation Smile (OS) has placed considerable emphasis on improving the quality of care delivered to patients in LMICs. By establishing an outcome evaluation program among other interventions, OS has achieved measurable improvements in complication rates and post-operative follow-up. The founding of the Guwahati Comprehensive Cleft Care Center in India has been pivotal to the success of OS's work in this field and is the evidence of the impact that an optimized model of care delivery can make. Here we describe OS's efforts to improve the quality of care delivered to patients in LMICs with a focus on the organization's work in India.

**Keywords:** cleft lip and palate, low and middle-income countries, global surgery,

Clefts of the lip and palate are among the most common congenital deformities, occurring in about 1 in 500 to 1 in 1000 births worldwide [1]. In many areas of the world, the number of patients with clefts far exceeds the capacity of the healthcare system to treat them. Consequently, only a fraction of the affected population receives treatment, resulting in a global backlog of over 600,000 untreated patients, with 72,000 in India alone [2]. Clefts of the lip and palate can have devastating consequences. Children with cleft palate have difficulty with speech and feeding, which can lead to malnourishment and developmental delay [3, 4]. Children with cleft lip face social stigma and often have lower levels of education and greater unemployment compared to their peers, despite having normal intellectual capacity [5–7]. The global response to this problem over the past several decades has been immense. Charitable organizations have provided more than 1 million free cleft repair surgeries and trained local surgeons to perform these procedures [8].

Historically, the success of these outreach efforts has been measured by the number of patients served, and little emphasis has been placed on the quality of care delivered. Research related to surgical missions is needed but seldom performed, often due to limited funding, manpower, and time [9, 10]. The logistical challenges of

*Carolina Restrepo and Alex Campbell*

models of health care delivery, quality improvement

**Chapter 3**


**1. Introduction**

Feynman was the first to present the idea of quantum computing, that is, to directly use the state of microscopic particles to represent quantum information, which is considered to be the early prototype of the concept of quantum computing [1]. Subsequently, Deutsch formalized the concept of quantum computing, proposed the idea of a quantum Turing machine, and designed the first quantum parallel algorithm, which exhibited excellent performance beyond classical computing [2]. The proposal of Shor's algorithm caused researchers to realize that quantum computing had a natural parallel processing capability, which could introduce many disruptive technological innovations. Shor's algorithm states that a large number can be decomposed into the product of two prime factors in polynomial time. This greatly challenged the RSA (Rivest–Shamir–Adleman) encryption system, thus indicating that the RSA encryption system had been cracked in theory [3,4]. Grover's search algorithm convinced researchers of the power of quantum computing. Compared with the traditional search method, this algorithm can achieve the acceleration effect of square level [5]. Therefore, many improved Grover search algorithms have been proposed [6–10]. Meanwhile, quantum-inspired algorithms have also been proposed that can be simulated by classical computing [11–16]. Moreover, the quantum algorithm has been applied to solve linear systems of equations, which introduced new ideas for solving linear equations. This algorithm is also called the *HHL* algorithm [17]. The *HHL* algorithm has been widely used, and its improved algorithms have been continuously proposed [18–20]. Recently, quantum algorithms have been applied to solve differential equations [21–24]. A series of quantum computing technologies, such as quantum Fourier transform [25], quantum phase estimation [26], and the *HHL* algorithm, are called quantum basic linear algebra assembly [27]. At present, quantum computing has been widely used in cryptography, quantum simulation, machine learning, and other fields and shows a strong ability and great potential.

**Citation:** Zhao, T.; Yang, T.; Chi, Y. Quantum Weighted Fractional Fourier Transform. *Mathematics* **2022**, *10*, 1896. https://doi.org/10.3390/ math10111896

Academic Editors: Fernando L. Pelayo and Mauro Mezzini

Received: 3 May 2022 Accepted: 31 May 2022 Published: 1 June 2022

**Publisher's Note:** MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

**Copyright:** © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/).

73

The Fourier transform plays an important role in the design of quantum algorithms, but little is known about the quantum algorithms of the fractional Fourier transform (*FRFT*). The initial definition of the *FRFT* was proposed in [28]. Its application provides a convenient technique for solving certain classes of ordinary and partial differential equations, which arise in quantum mechanics from classical quadratic Hamiltonians. The theoretical research of the *FRFT* has developed rapidly, and various definitions have been proposed, such as eigenvalue *FRFT* [29], weighted *FRFT* [30], and sampling *FRFT* [31]. These definitions are widely used in various fields of signal processing. So far, little is known about the reports and studies on the quantum fractional Fourier transform (*QFRFT*). The main reason is that the design of quantum algorithms should satisfy unitarity, and some FRFTs do not include unitarity. Thus, a quantum pseudo-fractional Fourier transform (*QPFRFT*) was proposed [32], and the authors showed that there was no *QFRFT*. However, we present a reformulation of the weighted fractional Fourier transform (*WFRFT*) and prove its unitarity, whereupon a quantum weighted fractional Fourier transform (*QWFRFT*) is proposed.

The remainder of this paper is organized as follows. The preliminary knowledge is described in Section 2. The unitarity of the WFRFT is proved in Section 3. Section 4 presents the *QWFRFT*. Finally, the conclusions are presented in Section 5.


*Article* **Optimization of Oleuropein and Luteolin-7-O-Glucoside Extraction from Olive Leaves by Ultrasound-Assisted Technology**

### **Antonio Lama-Muñoz \*, María del Mar Contreras, Francisco Espínola, Manuel Moya, Inmaculada Romero and Eulogio Castro**

Department of Chemical, Environmental and Materials Engineering, University of Jaén, Campus Las Lagunillas, s/n, Building B3, 23071 Jaén, Spain

**\*** Correspondence<EMAIL_ADDRESS>Tel.: +34-953-211-860

Received: 29 May 2019; Accepted: 27 June 2019; Published: 28 June 2019

**Abstract:** The olive orchard cultivation in Mediterranean countries results in huge amounts of lignocellulosic biomass residues. One of the main residues are olive leaves. Olive leaves contain high concentrations of bioactive antioxidant compounds like oleuropein and luteolin-7-glucoside. The production of biactive compounds from olive leaves requires treatments capable of breaking the lignocellulosic structure. Current research focuses on use of inexpensive, quick, and not harmful to the environment treatments, searching a more simplified large-scale operation approach. Recently, advances in applied chemistry have led to possible new emerging industrial techniques like ultrasound-assisted extraction (UAE). This technology is a promising candidate as a green treatment solution for olive leaves utilization in a biorefinery. However, this application goes through prior optimization of technique and operating conditions. The goal of this study was to optimize the extraction of oleuropein and luteolin-7-glucoside from olive leaves through an investigation of the influence of key factors of ultrasound-assisted extraction using an experimental central composite design, in comparison with conventional Soxhlet extraction. The highest extraction efficiency and antioxidant capacity were obtained under optimal increment of temperature and amplitude conditions (40 ◦C and 30%, respectively). Values for oleuropein, luteolin-7-glucoside were 69.91 g/kg and 1.82 g/kg, respectively.

**Keywords:** antioxidants; bioactive compounds; biorefinery; experimental design; olive leaves; olive biomass; response surface methodology; ultrasound-assisted extraction; olive byproducts valorization


**5. Asymptomatic malaria infections and mass drug administration (MDA)**

One of the major problems to achieve malaria elimination is represented by the hidden parasite reservoir in the human host. Microscopy (and rapid diagnostic tests (RDTs)) underestimates by about half the prevalence of *Plasmodium* infection, and this difference is greatest in lowtransmission settings—many asymptomatic infections can persist for significant periods of time. The presence of *P. falciparum* gametocytes is positively associated with the absence of clinical symptoms and low asexual parasite densities; mosquitoes can become infected with gametocyte densities as low as 5 gametocytes/μl and theoretically as low as one gametocyte/μl—children with undetectable gametocytaemia by molecular methods were still observed to be infectious to mosquitoes [17]. To accelerate achieving malaria elimination, the human reservoir of infection needs to be tackled with new approaches. There is a growing interest in MDA of at-risk populations or in malaria hot-spot areas with an effective antimalarial to reduce the parasite reservoir in human host [18]. MDA aims to provide full post-treatment courses to the whole population to clear asymptomatic infections and provide posttreatment prophylaxis to prevent reinfection. The use of MDA is recommended in areas approaching interruption of transmission, with good access to treatment, effective vector control, and surveillance systems, ensuring a minimal risk of reintroduction of infection [19]. MDAs have been conducted using a variety of drug regimens at different dosages, timings, and frequency. There is evidence of substantial but short-lived reduction in *P. falciparum* parasite carriage [20]. In Zambia, a cluster-randomized control trial implemented in a population of 330,000 individuals, distributed in 56,000 households, compared MDA with DP (2 rounds), at the household level (DP to all members of household with at least a RDT-positive individual) and standard control measures (case management, LLIN, indoor residual spraying (IRS), and intermittent preventive treatment during pregnancy). MDA decreased significantly malaria prevalence and incidence in low (malaria prevalence <10%) but not in high (malaria prevalence ≥10%) transmission areas [21]. With the growing awareness of heterogeneity and clustering in transmission, MDA approaches have been modified by systematic (mass screening and treatment) or focused (focal screening and treatment) screening and treatment of populations in defined geographical areas. Reactive case detection, i.e., screening and treating positive contacts in response to a clinical event, has been tested and implemented in some countries [22–25]. However, its impact has been variable as it is affected by the sensitivity of the diagnostic tool and the radius of intervention around a clinical case [26–29].

trial in a country with high coverage of standard control interventions and substantial residual

Malaria Elimination: Challenges and Opportunities http://dx.doi.org/10.5772/intechopen.77092 7

In conclusion, achieving universal access to standard control interventions, namely, case management, LLIN, IRS, seasonal malaria chemoprevention, and intermittent preventive treatment for pregnant women, remains a priority. It is essential to contain emerging drug resistance in malarial parasite and insecticide resistance in mosquito vector species. There is a dire need of additional new interventions to accelerate interruption of transmission. These should be evaluated and rapidly integrated within the standard control activities. Most of these should be implemented at the community level, and it will be important to actively involve the local populations to reach high coverage. Finally, political and financial supports should be maintained and even increased; current financial support is less than half of that estimated to reach the 2030 targets of the WHO global technical strategy for malaria [1].

Medical Research Council Unit, The Gambia and London School of Hygiene and Tropical

[1] WHO. World malaria report. Geneva: World health organization; 2017. Licence: CC BY-NC-SA 3.0 IGO. http://apps.who.int/iris/bitstream/handle/10665/259492/9789241565523 eng.pdf;jsessionid=DBAAA4A8F412049581DEC3652C2EE8E5?sequence=1; 12 December

[2] WHO. Global Malaria Programme. A framework for malaria elimination. Geneva: World Health Organization; 2017. http://apps.who.int/iris/bitstream/handle/10665/

[3] Alonso PL, Brown G, Arevalo-Herrera M, Binka F, Chitnis C, et al. A research agenda to underpin malaria eradication. PLoS Medicine. 2011;**8**:e1000406. DOI: 10.1371/journal.

[4] The malERA Refresh Consultative Panel on Insecticide and Drug Resistance. An updated research agenda for insecticide and drug resistance in malaria elimination and eradica-

tion. PLoS Medicine. 2017;**14**:e1002450. DOI: 10.1371/journal.pmed.1002450

254761/9789241511988-eng.pdf?sequence=1; 12 December 2017

malaria transmission.

**6. Conclusions**

**Author details**

**References**

2017

pmed.1000406

Umberto D'Alessandro

Medicine, Fajara, The Gambia

Address all correspondence to<EMAIL_ADDRESS>
The antimalarial treatment administered during MDA campaigns could be complemented by single low-dose of primaquine, an 8-aminoquinoline that is able to clear mature *P. falciparum* gametocytes [30], and/or ivermectin, a systemic endectocidal drug that can be administered safely to both humans and animals but proven toxic to *Anopheles* mosquitoes when they take a blood meal from a host that has recently received the drug [31, 32]. Primaquine may cause a dose-dependent hemolysis, mainly in individuals with deficiency of the enzyme glucose 6-phosphate dehydrogenase (G6PD) in red blood cells [33], and this has slowed down its implementation. Nevertheless, a single low-dose of primaquine can significantly reduce gametocyte carriage in both symptomatic [33] and asymptomatic [34] individuals and reduces onward transmission from man to vector [35]. Ivermectin can be safely administered with an ACT [36, 37] and has been used widely against parasitic diseases in humans, with record of more than 2 billion doses in MDA campaigns against onchocerciasis and lymphatic filariasis. In Burkina Faso, Liberia, and Senegal, one round of MDA with ivermectin at the standard dose of 150 μg/kg decreased substantially *An. gambiae* survival for 6 days and reduced the proportion of sporozoite-positive (infectious) mosquitoes for 2 weeks [38]. However, evidence of ivermectin as an additional tool to decrease malaria transmission is limited and needs to be further quantified, possibly by a cluster randomized trial in a country with high coverage of standard control interventions and substantial residual malaria transmission.


**5. Diagnosis**

According to the American College of Gastroenterology (ACG), *H. pylori* testing is indicated in the following cases: [6].

All patients with active PUD

A complete history and physical examination are essential for identifying risk factors, symptoms, and signs associated with *H. pylori* infection [7]. These can include epigastric pain lasting for more than 4 weeks, or alarm symptoms like unexplained weight loss, progressive dysphagia, odynophagia, recurrent vomiting, abdominal pain, iron deficiency anemia (IDA), jaundice, gastrointestinal (GI) bleeding, and a family history of GI cancer. Following a thorough history and physical exam, appropriate diagnostic investigations should be recommended based on the assessment. There are a variety of invasive and non-invasive techniques to appropriately make the diagnosis of an *H. pylori* infection. Patients under age 55 and without alarm symptoms can be advised to go for noninvasive tests. Following are the different diagnostic investigations available in different clinical settings:


**6.5 Strategies in people with diabetes to reduce the risk of HF**

The 2019 ESC-EASD guidelines recommend regular microalbuminuria and eGFR screening to identify patients at high risk of renal dysfunction or future CVD. On the other hand, the Standards of Care 2021 from the ADA [126], and the 2019 ESC-EASD guidelines [114] recommend a BP target of <130/80 mmHg (but not <120 mmHg).

Moreover, even though the 2021 ESC guidelines [91] do not recommend a target, the 2022 AHA/ACC/HFSA guidelines [120] do recommend a more stringent target of systolic BP of <120 mmHg in individuals with diabetes at CV risk since hypertension control is associated with a lower HF risk. It is worth noting that masked hypertension (meaning only home, but not office BP levels are hypertensive) [127] is common in T2D patients [128], making out-of-office BP monitoring a viable screening method for this clinical condition [129, 130]. Diabetic and hypertensive patients should have their ECGs checked at rest to identify silent MI, which happens in 4% of diabetic patients and adds an insult to HF [114]. Additionally, for pre-diabetics and hypertensive patients with diabetes, lifestyle adjustments and the use of RAAS blockers as first-line therapy for BP management are advised [114]. RAAS blockers also diminish the incidence of new-onset diabetes and the risk of sudden cardiac death in HFrEF patients [114]. Aside from hypertension, a higher body mass index is thought to be a risk factor for HF, which is why the ESC recommendations for 2021 [91] propose that obesity should be controlled to avoid or delay the onset of HF.


**3. Ways of biofortification**

Because traditional therapies are ineffective, biofortification has been advocated as a long-term alternative for increasing mineral nutrition [17]. Biofortification is a method that improves both mineral content and bioavailability in the edible parts of staple crops. The former can be accomplished through agronomic intervention, plant breeding, or genetic engineering, whereas mineral bioavailability can only be influenced through plant breeding and genetic engineering (**Figure 2**).


Meet the editors

Wei Wu is assistant to the Dean at the School of Public Health, and Professor and Associate Department Chair of the Department of Toxicology at Nanjing Medical University, China, where he received his Ph.D. in toxicology in 2012. He was a guest researcher at the National Institute of Environmental Health Sciences (NIEHS) between 2017 and 2018. He is the PI of the State Key Laboratory of Reproductive Medicine and

the PI of the Key Laboratory of Modern Toxicology at the Ministry of Education. Dr. Wu is a member of several national and international societies in the fields of human reproduction and toxicology and has received awards from many national societies for the originality and quality of his projects. Dr. Wu has authored 78 peer-reviewed papers in international journals. He has edited four books and collaborated on 10 books, 11 patents and in the organization of three international conferences. He is a reviewer for 104 international journals.

Panagiotis Tsikouras is a specialist in obstetrics and gynecology, perinatal medicine, and contraception at the School of Medicine, Democritus University of Thrace, Greece. He is also head of the Family Planning Centre and Gynecological Cytology Laboratory at the same university. Dr. Tsikouras is a fellow of the International Academy of Clinical and Applied Thrombosis/Hemostasis. His scientific activities focus on pediatric

and adolescent medicine, gynecological oncology, and high-risk pregnancies. He is a reviewer for several international journals and has numerous scientific publications to his credit, including papers and book chapters. He has also contributed to international and national guidelines on coagulation and thrombosis in obstetrics and gynecology.

Qiuqin Tang is an associate chief physician of the Women's Hospital of Nanjing Medical University (Nanjing Maternity and Child Health Care Hospital). Her research interests include genetic and epigenetic risk factors in reproductive and developmental health. She has authored 29 papers in international journals such as *EBioMedicine*, *Environmental Pollution*, *Clinical Epigenetics*, *Molecular Human Reproduction*, and *European*

*Journal of Endocrinology*. She has collaborated on four books and three patents. She is the editor-in-chief of the *Journal of Woman's Reproductive Health*.

Werner Rath is a specialist in obstetrics and gynecology, gynecological oncology, perinatal medicine and hemostaseology. Currently a professor of gynecology and obstetrics at Kiel University, he previously led the Department of Gynecology and Obstetrics at University Hospital, Aachen. He holds an honorary doctorate from the University of Alexandroupolis. He has received the Staude-Pfannenstiel Award of the North German

Society of Gynaecology & Obstetrics, the Maternity Award of the German Society

of Perinatal Medicine, Berlin, and the Harald-Bräutigam Award, Hamburg. His citable research publications date from 1977. He is a reviewer for numerous journals and, in addition to his publications and 13 book chapters, was chief editor of the German *Journal of Obstetrics & Gynaecology* (*Geburtshilfe und Frauenheilkunde*).

Georg-Friedrich von Tempelhoff, FCATH, is a specialist in obstetrics and gynecology, gynecological oncology, perinatal medicine and hemostaseology. He is head of the Department of Gynecology and Obstetrics at St. Vinzenz Hospital, Hanau and head of the Institute for Thrombosis and Coagulation Disorders in Gynecology and Obstetrics, Hausen. He was educated at the Justus-Liebig University in Giessen, receiving his doctorate in medicine in 1993.

He became a consultant in obstetrics and gynecology in 1996. He completed his habilitation at the Johannes-Gutenberg University Mainz in 2007 and was appointed extracurricular professor at the Julius-Maximilians-Universität Würzburg in 2015. He is a fellow of the International Academy of Clinical and Applied Thrombosis / Hemostasis. His research focus is on hemostaseological and hemorheological disorders in women with miscarriages, pre-eclampsia and thrombosis. He is a reviewer for numerous journals and, in addition to his publications and book chapters, has contributed to international and national guidelines on coagulation and thrombosis in gynecology.

Nikolettos Nikos has been a professor of obstetrics, gynecology and assisted reproduction at Democritus University of Thrace, since 2018, and has managed the Obstetrics and Gynecology Clinic since 2019. From 2007 to 2011 he was a director of the In-Vitro Fertilization Unit of the University Regional General Hospital, Alexandroupolis. From 2012 to 2018 he was director of the Laboratory of Reproductive Physiology and Artificial Fertilization.

He has more than 100 publications in international journals and has delivered many lectures by invitation to international and Greek conferences.

## Contents


*3.11. Determination of Molecular Weight*

The molecular weight of CamPhoD was determined using gel filtration on a calibrated Superdex G200 PG (GE Healthcare Life Sciences, Amersham Place, Little Chalfont, Buckinghamshire, HP7, 9NA, UK) column (105 × 2 cm) and polyacrylamide gel electrophoresis under denaturing conditions (sodium dodecyl sulfate polyacrylamide gel electrophoresis) using the Lammley method [61].


**Abstract**

Stroke remains a major public health issue and the second leading cause of death worldwide. The Hippocratic Corpus used the word apoplexy to describe a person collapsing while retaining pulse and respiration. This is believed to be the first written description of stroke. The theories of what caused stroke evolved over the years. When autopsies were performed stroke was attributed to emboli and thrombi formation. Carotid endarterectomies (CEA) were then performed for the treatment of stroke. Originally CEA were seen with skepticism but the North American Symptomatic Carotid Endarterectomy trial (NASCET) and the European Carotid Surgery trial (ECS) helped restore their efficacy in the management of ischemic stroke. A milestone in the management of ischemic stroke was the use of intravenous tissue plasminogen activator (tPA). Secondary to the limitations of the use of tPA other avenues were sought which included intraarterial recombinant prourokinase and mechanical thrombectomy. The field of mechanical thrombectomy continues to be rapidly changing and evolving. Various randomized controlled trials and meta-analysis have been conducted in order to evaluate who will benefit from mechanical thrombectomies, the timing, the best device to use and the role of combining this intervention with the administration of intravenous tPA.

**Keywords:** Ischemic stroke, history of stroke, carotid endarterectomy, intravenous tissue plasminogen activator, endovascular mechanical thrombectomy


**5. Conclusion**

In this chapter, we introduced a new approach to determine the best solution out of *m* candidates. To do this, first, we defined the belief of selecting the best solution and explained how to model the problem by the Bayesian analysis approach. Second, we clarified the approach by which we improved the beliefs, and proved that it converges to detect the best solution. Next, we proposed a decision-making strategy using dynamic programming approach in which there were a limited number of decision-making stages.


*3.7. Electron Microscopy Analysis of Precipitate Structure Evolution*

TEM analyses were done in order to understand how the mechanical properties of Mg samples are related to microstructural features, in particular, to different shapes and densities of precipitates that form during heat treatments. The samples were chosen based on the significance of microhardness results shown in Figure 10, for the alloy Mg5Zn0.3Ca, in the following three states:

The three precipitate states reveal differences in morphology, size and density of precipitates in the range of 3–100 nm. Primary precipitates, as scattered residuals after annealing, did not show any substantial evolution during the processing history; see Figure 16—first row, SEM 200×.

**Figure 16.** SEM (upper two rows) and STEM (bottom row) images of Mg5Zn0.3Ca samples (**a**) after HPT-processing; (**b**) after HPT and additional heat treatment at 100 ◦C for 24 h; and (**c**) after HPT and additional heat treatment at 125 ◦C for 24 h. HPT-processing was carried out by 0.5 rotations under 4 GPa pressure at RT.

Precipitates' structures have been analyzed in all three states (Figure 16—second row, SEM about 10,000×), namely: polyhedral precipitates (P1), elongated precipitates (P2) and thinner phases. The thinner phases are at the boundaries of grains, or follow their contours. P1 and P2 do not change at all during processing, and can be observed in all states (Figure 17). P1 and P2 contain Zn and Ca, as seen in exemplary EDS maps of P1 precipitates (Figure 18).

**Figure 17.** STEM image of Mg5Zn0.3Ca, HPT-processed at 0.5 rotations, showing P1, P2 and thinner precipitates (blue arrows). The TEM image in the top left corner is a close-up of the P1 precipitate. Thinner precipitates are located at the grain boundaries, following their contours.

**Figure 18.** EDS maps of a P1 precipitate in a Mg5Zn0.3Ca sample. These precipitates contain Zn (green) and Ca (blue); Mg (red) was mainly observed in the surrounding matrix. O (pink) appeared in all sample areas; it most likely came from the atmospheric oxidation of prepared TEM foils.

Besides P1, P2 and thinner phases, further precipitates were observed in the heat-treated samples (Figure 16—third row, STEM, about 200,000×). P3-precipitates could be observed in both the heat -treated samples (Figure 19). In the sample heat treated at 100 ◦C, the P3-precipitates were located at the boundaries between grains and had a size of 20-50 nm. In the sample heat treated at 125 ◦C, those phases grew in size up to 100 nm. Additionally, ultrafine P4 precipitates (Figure 19) with a size less than 10 nm and up to 50 nm appeared. They were oriented normal to <0001> direction, but further verifications are required. These oriented platelets were not observed in the other two conditions.

**Figure 19.** TEM images of the HPT-processed (0.5 rot) and heat-treated Mg5Zn0.3Ca sample (**a**) at 100 ◦C for 24 h and (**b**) HPT processed and heat treated at 125 ◦C for 24 h showing P3 and P4 precipitates.