*Article* **Genomic Sequencing and Comparison of Sacbrood Viruses from** *Apis cerana* **and** *Apis mellifera* **in Taiwan**

**Ju-Chun Chang 1,2, Zih-Ting Chang 1, Chong-Yu Ko 1, Yue-Wen Chen 1,\* and Yu-Shin Nai 2,\***

1 Department of Biotechnology and Animal Science, National Ilan University, Yilan 260, Taiwan;

<EMAIL_ADDRESS>(J.-C.C<EMAIL_ADDRESS>(Z.-T.C<EMAIL_ADDRESS>(C.-Y.K.)

2 Department of Entomology, National Chung-Hsing University, Taichung 402, Taiwan **\*** Correspondence<EMAIL_ADDRESS>(Y.-W.C<EMAIL_ADDRESS>(Y.-S.N.)

**Abstract:** Sacbrood virus (SBV) was the first identified bee virus and shown to cause serious epizootic infections in the population of *Apis cerana* in Taiwan in 2015. Herein, the whole genome sequences of SBVs in *A. cerana* and *A. mellifera* were decoded and designated AcSBV-TW and AmSBV-TW, respectively. The whole genomes of AcSBV-TW and AmSBV-TW were 8776 and 8885 bp, respectively, and shared 90% identity. Each viral genome encoded a polyprotein, which consisted of 2841 aa in AcSBV-TW and 2859 aa in AmSBV-TW, and these sequences shared 95% identity. Compared to 54 other SBVs, the structural protein and protease regions showed high variation, while the helicase was the most highly conserved region among SBVs. Moreover, a 17-amino-acid deletion was found in viral protein 1 (VP1) region of AcSBV-TW compared to AmSBV-TW. The phylogenetic analysis based on the polyprotein sequences and partial VP1 region indicated that AcSBV-TW was grouped into the SBV clade with the AC-genotype (17-aa deletion) and was closely related to AmSBV-SDLY and CSBV-FZ, while AmSBV-TW was grouped into the AM-genotype clade but branched independently from other AmSBVs, indicating that the divergent genomic characteristics of AmSBV-TW might be a consequence of geographic distance driving evolution, and AcSBV-TW was closely related to CSBV-FZ, which originated from China. This 17-amino-acid deletion could be found in either AcSBV or AmSBV in Taiwan, indicating cross-infection between the two viruses. Our data revealed geographic and host specificities between SBVs. The amino acid difference in the VP1 region might serve as a molecular marker for describing SBV cross-infection.

**Keywords:** sacbrood virus; sacbrood disease; *Apis cerana*; *Apis mellifera*


**8. Operative treatment**

Operative treatment of Dupuytren's disease is offered in patient with contractures of >30° at the MCP joint and any functionally bothersome PIP contracture. The goal of surgical treatment is to return full extension of the involved digits via various surgical techniques involving either incising or excising the diseased fascia. Dupuytren originally described an open palmar fasciotomy technique in 1831 and this was later popularized in 1964 by McCash et al. as the open-palm technique [6, 66]. The open-palm technique involved a transverse incision across the distal palmar crease followed by incising any Dupuytren cords. Multiple surgical methods have since been described and include open fasciotomy, segmental fasciectomy, limited fasciectomy, and dermofasciectomy. These techniques range from being minimally invasive to radical excision of the diseased tissue. It is important to consider the severity of contractures, extent of correction, and risk factors for recurrence in addition to protecting soft tissues when choosing the optimal surgical treatment. Surgery is ultimately an elective form of treatment and should prompt a conversation with patients regarding the risk and benefits of surgery as well as their functional goals.

#### **8.1. Soft tissues**

Proper handling of soft tissue is a key principle of surgical treatment of Dupuytren's contractures. Adequate exposure of the cord must be balanced with protecting the neurovascular bundles, providing adequate wound coverage, limiting the risk of skin necrosis, and avoiding secondary contractures from longitudinal scarring. Multiple skin incisions have been described to address these issues and include: transverse incisions in the palm and digit, a Bruner incision, a Bruner incision with V-Y advancement flaps, curved incisions, and a longitudinal incision closed with z-plasties (**Figure 3**). Midline incisions provide the benefit of potentially avoiding the neurovascular bundles, however, Dupuytren's disease distorts regular anatomy and can tether the cord toward the midline. Curved or zig-zag incisions help avoid secondary contractures from longitudinal scars. Despite the many variations in incisions, the Bruner incision seems to be the most commonly used.

Skin closure is another important aspect of treating Dupuytren's disease. The open-palm technique described by McCash left the skin incisions open to decrease hematoma formation and allowed for secondary healing with good results [66]. Today, most incisions are closed primarily, however, increased skin tension after closure has been correlated with elevated recurrence rate (**Figure 4**). Citron and Hearnden randomized patients undergoing fasciotomies and reported a 50% recurrence rate in the group with transverse incisions closed primarily compared with a 15% rate in patients with longitudinal incisions with a z-plasty closure [67]. Another study compared Bruner's incision with direct closure to a longitudinal incision with a z-plasty closure for fasciectomy and reported no difference between the two methods [68]. Special attention should be given to skin tension during closure and a transpositional flap such as a z-plasty should be utilized if needed.

#### **8.2. Open fasciotomy**

Open fasciotomy includes a variety of surgical techniques for treating Dupuytren's contractures by incising the contracted cord without removing the diseased fascia. Many of the modern fasciotomy techniques are modifications of the methods originally described by Dupuytren and

> McCash et al. [6, 66]. Various types of incisions may be utilized to access the cord, but once the cord is visualized and incised, the digit is extended until straight. In some cases, additional incisions may be required at different levels along the cord in order to fully extend the digit. Unlike PNF, an open fasciotomy procedure provides the benefit of direct visualization to protect neurovascular structures and can often be accomplished through small incisions. In addition, it minimizes the potential morbidity sometimes seen in other techniques which excise the diseased fascia. Still, excision techniques are often preferred over an open fasciotomy for their ability to remove the diseased fascia which may aid in preventing regrowth of the cord and recurrence. This may be particularly true in patients with severe contractures. Stewart et al. retrospectively reviewed a series of patients who had open fasciotomies and reported a reoperation rate of 13.2% at 46 months with patients who initially required three level fasciotomies having worse recurrence [69]. Another study assessed 16 patients with Tubiana stage III and IV

> **Figure 4.** A post-operative image of a patient after a limited fasciectomy for a Dupuytren's contracture of the small and

Dupuytren's Disease

71

http://dx.doi.org/10.5772/intechopen.72759

ring finger. The Brunner's incision was closed primarily with nylon sutures.

contractures and reported a higher recurrence rate of 37.5% at 5–8 year follow-up [70].

Fasciectomy is the most common surgical treatment for Dupuytren's contractures. The technique is based on the concept that the remaining diseased fascia may proliferate and lead to recurrence. Multiple methods of fasciectomies have been described and range from a segmental fasciectomy, where a portion of the fascial cord is removed, to a radical fasciectomy where skin and diseased fascia are excised. A segmental fasciectomy is usually completed through small incisions where segments of the cord are excised until the finger straightens. No attempt

**8.3. Fasciectomy**

**Figure 3.** The figure outlines types of incisions and closures options for treating Dupuytren's disease. Thumb: longitudinal incision with z-plasty closure. Index finger: Brunner incision. Middle finger: curvilinear incision. Index finger: V-Y incision. Small finger: transverse incisions utilized in the McCash open palm technique.

**Figure 4.** A post-operative image of a patient after a limited fasciectomy for a Dupuytren's contracture of the small and ring finger. The Brunner's incision was closed primarily with nylon sutures.

McCash et al. [6, 66]. Various types of incisions may be utilized to access the cord, but once the cord is visualized and incised, the digit is extended until straight. In some cases, additional incisions may be required at different levels along the cord in order to fully extend the digit. Unlike PNF, an open fasciotomy procedure provides the benefit of direct visualization to protect neurovascular structures and can often be accomplished through small incisions. In addition, it minimizes the potential morbidity sometimes seen in other techniques which excise the diseased fascia. Still, excision techniques are often preferred over an open fasciotomy for their ability to remove the diseased fascia which may aid in preventing regrowth of the cord and recurrence. This may be particularly true in patients with severe contractures. Stewart et al. retrospectively reviewed a series of patients who had open fasciotomies and reported a reoperation rate of 13.2% at 46 months with patients who initially required three level fasciotomies having worse recurrence [69]. Another study assessed 16 patients with Tubiana stage III and IV contractures and reported a higher recurrence rate of 37.5% at 5–8 year follow-up [70].

#### **8.3. Fasciectomy**

Bruner incision, a Bruner incision with V-Y advancement flaps, curved incisions, and a longitudinal incision closed with z-plasties (**Figure 3**). Midline incisions provide the benefit of potentially avoiding the neurovascular bundles, however, Dupuytren's disease distorts regular anatomy and can tether the cord toward the midline. Curved or zig-zag incisions help avoid secondary contractures from longitudinal scars. Despite the many variations in incisions, the

Skin closure is another important aspect of treating Dupuytren's disease. The open-palm technique described by McCash left the skin incisions open to decrease hematoma formation and allowed for secondary healing with good results [66]. Today, most incisions are closed primarily, however, increased skin tension after closure has been correlated with elevated recurrence rate (**Figure 4**). Citron and Hearnden randomized patients undergoing fasciotomies and reported a 50% recurrence rate in the group with transverse incisions closed primarily compared with a 15% rate in patients with longitudinal incisions with a z-plasty closure [67]. Another study compared Bruner's incision with direct closure to a longitudinal incision with a z-plasty closure for fasciectomy and reported no difference between the two methods [68]. Special attention should be given to skin tension during closure and a transpositional

Open fasciotomy includes a variety of surgical techniques for treating Dupuytren's contractures by incising the contracted cord without removing the diseased fascia. Many of the modern fasciotomy techniques are modifications of the methods originally described by Dupuytren and

**Figure 3.** The figure outlines types of incisions and closures options for treating Dupuytren's disease. Thumb: longitudinal incision with z-plasty closure. Index finger: Brunner incision. Middle finger: curvilinear incision. Index finger: V-Y incision.

Small finger: transverse incisions utilized in the McCash open palm technique.

Bruner incision seems to be the most commonly used.

flap such as a z-plasty should be utilized if needed.

**8.2. Open fasciotomy**

70 Essentials of Hand Surgery

Fasciectomy is the most common surgical treatment for Dupuytren's contractures. The technique is based on the concept that the remaining diseased fascia may proliferate and lead to recurrence. Multiple methods of fasciectomies have been described and range from a segmental fasciectomy, where a portion of the fascial cord is removed, to a radical fasciectomy where skin and diseased fascia are excised. A segmental fasciectomy is usually completed through small incisions where segments of the cord are excised until the finger straightens. No attempt is made to removal the complete cord. The most widely used technique is a limited fasciectomy and is considered the gold standard in the operative treatment of Dupuytren's contractures (**Figure 5**). The technique involves carefully exposing the diseased fascia from its proximal to distal end and excising it from the surrounding soft tissue (**Figure 6**). It differs from a radical fasciectomy by removing only the diseased fascia and leaving normal fascia, subcutaneous tissue, and the dermis intact. Radical fasciectomy advocated by McIndie and Beare involved extensive removal of nearby tissue and skin and required a skin graft [71]. However, the technique largely fell out of favor due to a higher complication rate without a reduction in recurrence. A dermatofasciectomy is similar to a limited fasciectomy but involves excising the overlying skin and often requires a skin graft for coverage. Advocates of this technique report disease-forming cells may be left in the overlying soft tissue leading to recurrence and selectively use it in patients at higher risk for recurrence. Despite the variety of techniques, fasciectomies require meticulous dissection to avoid injuring neurovascular bundles which may be displaced by a contracted cord. In addition, maintaining hemostasis is important to prevent hematoma formation which can compromise healing. Overall, fasciectomies offer the benefit of removing more diseased fascia but are accompanied by increased morbidity related to a more extensive exposure.

#### **8.4. Post-operative protocol**

Most surgeons utilize some form of rehabilitation to prevent further contractures and maintain ROM. After surgery, patients are typically immobilized in an extension-based splint for 2–3 days. Wounds are closely monitored following surgery to ensure adequate healing and to identify any barriers to healing such as infection, hematoma, or skin necrosis. Patients are often referred to a hand therapist within the first week of surgery for wound care, scar management, range of motion exercises, and splinting techniques to prevent contracture formation. A variety of splinting protocols and types of orthoses have been described. In general, protocols may involve static or dynamic splinting and be utilized at different periods of the day. Despite the widespread use of post-operative splinting, studies have found no strong evidence they are effecting in preventing loss of extension or recurrence. Collis et al. randomized patients to night

time extension orthoses and hand therapy or hand therapy alone and reported no differences between the two groups in terms of active ROM or hand function [72]. A similar study found night time splinting offered no benefit in terms of ROM and function at 1 year after surgery and recommended splinting should only be utilized when extension deficits occur [73]. Overall, less invasive procedures allow for earlier rehabilitation and a shorter recovery period. In addition, wound healing often dictates how fast a patient can progress following surgery. A segmental aponeuroectomy may involve a 2–3 week recovery period whereas a fasciectomy may involve

**Figure 6.** An intraoperative image of a patient undergoing a limited fasciectomy for a Dupuytren's contracture of the ring finger. (A) A Brunner's incision was utilized to exposed the disease tissue. The longitudinal band of the palmar aponeurosis is being elevated. (B) The diseased fascia was meticulously elevated from proximal to distal. (C) The diseased

Dupuytren's Disease

73

http://dx.doi.org/10.5772/intechopen.72759

Overall, surgery is an effective method of treating Dupuytren's contractures and improving patients' hand function. In a survey of over 1100 patients who underwent surgical treatment 75% reported almost full or full correction of their contracture [74]. Zyluk et al. reported patients had significantly improved hand function as measured by the Disabilities of the Arm, Shoulder, and Hand (DASH) questionnaire after undergoing a subtotal fasciectomy [75]. In

a much longer recovery period.

fascia after removal from the digit.

**9. Patient outcomes**

**Figure 5.** An intraoperative image of a patient undergoing a limited fasciectomy for a Dupuytren's contracture of the small finger. A Brunner's incision was utilized to exposed the disease tissue. The Dupuytren's cord is being elevated by the forceps.

**Figure 6.** An intraoperative image of a patient undergoing a limited fasciectomy for a Dupuytren's contracture of the ring finger. (A) A Brunner's incision was utilized to exposed the disease tissue. The longitudinal band of the palmar aponeurosis is being elevated. (B) The diseased fascia was meticulously elevated from proximal to distal. (C) The diseased fascia after removal from the digit.

time extension orthoses and hand therapy or hand therapy alone and reported no differences between the two groups in terms of active ROM or hand function [72]. A similar study found night time splinting offered no benefit in terms of ROM and function at 1 year after surgery and recommended splinting should only be utilized when extension deficits occur [73]. Overall, less invasive procedures allow for earlier rehabilitation and a shorter recovery period. In addition, wound healing often dictates how fast a patient can progress following surgery. A segmental aponeuroectomy may involve a 2–3 week recovery period whereas a fasciectomy may involve a much longer recovery period.

### **9. Patient outcomes**

is made to removal the complete cord. The most widely used technique is a limited fasciectomy and is considered the gold standard in the operative treatment of Dupuytren's contractures (**Figure 5**). The technique involves carefully exposing the diseased fascia from its proximal to distal end and excising it from the surrounding soft tissue (**Figure 6**). It differs from a radical fasciectomy by removing only the diseased fascia and leaving normal fascia, subcutaneous tissue, and the dermis intact. Radical fasciectomy advocated by McIndie and Beare involved extensive removal of nearby tissue and skin and required a skin graft [71]. However, the technique largely fell out of favor due to a higher complication rate without a reduction in recurrence. A dermatofasciectomy is similar to a limited fasciectomy but involves excising the overlying skin and often requires a skin graft for coverage. Advocates of this technique report disease-forming cells may be left in the overlying soft tissue leading to recurrence and selectively use it in patients at higher risk for recurrence. Despite the variety of techniques, fasciectomies require meticulous dissection to avoid injuring neurovascular bundles which may be displaced by a contracted cord. In addition, maintaining hemostasis is important to prevent hematoma formation which can compromise healing. Overall, fasciectomies offer the benefit of removing more diseased fascia but are accompanied by increased morbidity related

Most surgeons utilize some form of rehabilitation to prevent further contractures and maintain ROM. After surgery, patients are typically immobilized in an extension-based splint for 2–3 days. Wounds are closely monitored following surgery to ensure adequate healing and to identify any barriers to healing such as infection, hematoma, or skin necrosis. Patients are often referred to a hand therapist within the first week of surgery for wound care, scar management, range of motion exercises, and splinting techniques to prevent contracture formation. A variety of splinting protocols and types of orthoses have been described. In general, protocols may involve static or dynamic splinting and be utilized at different periods of the day. Despite the widespread use of post-operative splinting, studies have found no strong evidence they are effecting in preventing loss of extension or recurrence. Collis et al. randomized patients to night

**Figure 5.** An intraoperative image of a patient undergoing a limited fasciectomy for a Dupuytren's contracture of the small finger. A Brunner's incision was utilized to exposed the disease tissue. The Dupuytren's cord is being elevated by the forceps.

to a more extensive exposure.

72 Essentials of Hand Surgery

**8.4. Post-operative protocol**

Overall, surgery is an effective method of treating Dupuytren's contractures and improving patients' hand function. In a survey of over 1100 patients who underwent surgical treatment 75% reported almost full or full correction of their contracture [74]. Zyluk et al. reported patients had significantly improved hand function as measured by the Disabilities of the Arm, Shoulder, and Hand (DASH) questionnaire after undergoing a subtotal fasciectomy [75]. In addition, patient's experienced a significant improvement in the total loss of extension from an average of 80 to 10°. Another study prospectively evaluated 90 patients undergoing a fasciectomy for a 60-degree or more deficit in total active extension and reported 81% patient satisfaction with function, 87% reaching functional ROM, and significantly improved DASH scores at 1 year follow up [76].

diathesis is a term coined by Hueston describing certain characteristics related to severe disease and increased recurrence [83]. Hindocha modified the criteria to include the following features within a Northern European population: male sex, <50 years old, bilateral disease, affected parent or sibling, and presence of Garrod's nodes and reported patients with all 5 features had a recurrence rate of 71% [84]. However, other studies have failed to demonstrate a significant correlation with recurrence among all 5 diathesis criteria. Van Rijssen reported only older age was found to delay recurrence after PNF and limited fasciectomy [62]. PIP joint contractures have an elevated recurrence rate after surgery compared to the MCP joint. Donaldson et al. reported 34.2% of fully corrected PIP joints experienced at least some loss of correction compared with 12.2% of MCP joints [80]. Patients with severe preoperative PIP contractures greater than 60°, incomplete correction, and poor post-operative compliance had significantly

Dupuytren's Disease

75

http://dx.doi.org/10.5772/intechopen.72759

Surgical management of recurrent disease is challenging as anatomic landmarks and tissue plans become difficult to distinguish. There is currently limited data regarding the preferred surgical treatment. Roush and Stern compared dermatofasciectomy with a skin graft, interphalangeal joint arthrodesis, and fasciectomy with local flaps among 19 patients with recurrence after a prior surgery [85]. The fasciectomy cohort was the only group to significantly maintain total active motion at final follow-up, but all three groups had similar patient reported outcomes. It is important patients are aware of the risk of recurrence prior to their initial surgery and personal factors which may increase their likelihood of

Despite good outcomes after surgical treatment of Dupuytren's contractures surgery is not without complications. A 20 year systematic review of complications by Denkler et al. reported an average major complication risk of 15% with complication rates ranging from 3.6 to 39.1% [86]. Specific complications after limited fasciectomy included the following: wound healing problems, 22.9%; flare reaction, 9.9%; complex region pain syndrome, 5.5%; nerve injury, 3.4%; infection, 2.4%; hematoma, 2.1%; and digit artery injury, 2%. Severe complications include tendon rupture or loss of the digit but are extremely rare. Patients with severe flexion contractures are more at risk of experiencing a complication [87]. Smoking and diabetes, however, has not been identified as an increased risk factor for wound healing problems after surgery [88]. Patients undergoing revision surgery for recurrence are most at risk for complications, especially neurovascular injuries due to scar tissue and loss of anatomic landmarks. Neurovascular injuries have been reported as high as 10 times more common in

Dupuytren's disease is a unique fibroproliferative disorder of palmar fascia likely resulting from a complex interplay of genetic and environmental factors. Despite extensive research,

worse recurrence [78].

recurrence.

**11. Complications**

revision surgeries for recurrence [86].

**12. Conclusion**

Further analysis of surgical treatment has lead authors to identify certain characteristic associated with better outcomes. Patients with MCP contractures are more likely to achieve full intra-operative correction (Donaldson). However, correction of PIP contractures has a stronger correlation to improved hand function when compared with correction of the MCP joint [77]. Surgical treatment has focused on PIP contractures to maximize intra-operative correction and improve functional outcomes. Surgeons have tried releasing the PIP capsule with a fasciectomy to improve PIP correction, but there is no strong data to support whether it is effective [78]. Zyluk et al. reported younger patients had a significantly greater functional improvement after surgery as measured by the DASH score [75]. Studies have also cited the extent of preoperative deformity, incomplete correction, and multiple involved digits as factors affecting post-operative functional outcomes [75, 78–80].

Despite multiple studies reporting good outcomes after surgery, there are limited randomized studies comparing outcomes after different operative techniques. Ullah et al. found no difference in ROM or recurrence in 79 patients randomized to either direct closure with z-plasty or firebreak skin grafting after a fasciectomy, however, patients with skin grafting had an increased incidence of hypoesthesia [81]. Van Rijssen et al. randomized patients to percutaneous needle fasciotomy and limited fasciectomy and reported the PNF group was significantly higher recurrence rate (76.8 vs. 20.9%, p < 0.001) and lower VAS satisfaction scores (6.2 vs. 8.3, p < 0.001) [62]. Further prospective, randomized studies reporting functional outcomes, complications, and recurrence rates are necessary to recommend any surgical procedure.

### **10. Recurrence**

Recurrence of a Dupuytren's contracture is a common event even after successful initial treatment. A systematic review analyzed 51 studies and reported recurrence rates ranged from 0 to 71% [82]. Furthermore, recurrence rates are difficult to assess as there is considerable variation in the criteria used to define recurrence. Some authors report the presence of any diseased tissue after treatment while others include only contractures necessitating re-operation. More recently, studies have tended to define recurrence as a 20–30° loss of extension in a successfully treated digit. A recent randomized study defined recurrence as a 20° reduction of total passive extension in a successfully treated digit and reported a 20.9% 5-year recurrence rate after limited fasciectomy [62].

Multiple studies have focused on identifying factors which may predispose patients to recurrence. The dramatic variability of recurrence rates may be due to the heterogeneity of the presentation of Dupuytren's itself, as many patients may have more aggressive biology associated with "Dupuytren's diathesis", whereas others may have more mild disease. Dupuytren diathesis is a term coined by Hueston describing certain characteristics related to severe disease and increased recurrence [83]. Hindocha modified the criteria to include the following features within a Northern European population: male sex, <50 years old, bilateral disease, affected parent or sibling, and presence of Garrod's nodes and reported patients with all 5 features had a recurrence rate of 71% [84]. However, other studies have failed to demonstrate a significant correlation with recurrence among all 5 diathesis criteria. Van Rijssen reported only older age was found to delay recurrence after PNF and limited fasciectomy [62]. PIP joint contractures have an elevated recurrence rate after surgery compared to the MCP joint. Donaldson et al. reported 34.2% of fully corrected PIP joints experienced at least some loss of correction compared with 12.2% of MCP joints [80]. Patients with severe preoperative PIP contractures greater than 60°, incomplete correction, and poor post-operative compliance had significantly worse recurrence [78].

Surgical management of recurrent disease is challenging as anatomic landmarks and tissue plans become difficult to distinguish. There is currently limited data regarding the preferred surgical treatment. Roush and Stern compared dermatofasciectomy with a skin graft, interphalangeal joint arthrodesis, and fasciectomy with local flaps among 19 patients with recurrence after a prior surgery [85]. The fasciectomy cohort was the only group to significantly maintain total active motion at final follow-up, but all three groups had similar patient reported outcomes. It is important patients are aware of the risk of recurrence prior to their initial surgery and personal factors which may increase their likelihood of recurrence.


**Acknowledgements**

**2.3. Future perspectives in defect engineering for superior electrocatalysis**

100 Graphene Oxide - Applications and Opportunities

low cost, environmental sustainability and high reproducibility.

reactions.

**3. Conclusions**

Immense research work has been done in the field of materials science toward the study of MOFs for highly efficient oxygen electrocatalysis [55]. Basically, MOFs are a class of composite materials with a unique and highly ordered nanoporous (pore sizes smaller than 100 nm) structure, which are designed from a self-assembly of inorganic metals and organic linkers through co-ordination bonds. From an electrochemical aspect, co-ordinated metals could contribute to an electronic effect as a consequence of the changes in platinum electronic structure, directly influencing catalyst activity. Thanks to abundant dual-metal active sites, superior electrocatalysts with ultra-low platinum content could be developed for alcohol electrocatalysis further improving specific activity beyond the conventional platinum alloy approach. Moreover, surface modification of rGO-supported platinum could lead to improved threedimensional nano- (or micro-) structures, resulting in synergistic effects for achieving faster mass and charge transport properties. In this context, proper synthetic procedures based on the controlled carbonization of the organic linkers (primarily containing heteroatoms) might serve as novel rational design strategies for the development of highly porous heteroatomdoped supported catalysts. Also, noteworthy is that such a synthetic method, especially for large-scale production, should meet the requirements for commercialization, which include

Last but not least, another contribution of the metal-organic framework process could be the *in situ* generation of metal oxide nanoparticles and their concomitant dispersion on rGO upon carbonization of the metal-organic complex. Thus, the deposited metal oxide nanoparticles could act as seeds for vacancy generation by etching the carbon─carbon network along the rGO-metal oxide interface. Size-controlled vacancies in the matrix of reduced graphene oxide are not only predicted to break the two-dimensional lattice symmetry, thereby tuning conduction mechanisms, but also act as trapping sites for heteroatom doping [56]. Therefore, in addition, heteroatom-doped porous and open structures on the basal planes together with the metal oxide nanoparticles might serve as novel active sites with high bifunctional activity. Precisely, a bifunctional effect has been associated to the presence of sites that aid in the dissociation of water to form surface hydroxides, which can readily oxidize strongly adsorbed reaction intermediates. Indeed, development of heteroatom-doped size-controlled vacancies could positively contribute toward the improvement of platinum activity in alcohol oxidation

As summarized in this mini-review, an ideal fuel-cell electrode should be porous, and possess high conductivity, accessible electrochemical surface sites, and improved charge and mass transfer pathways. Defect engineering, which involves manipulating the type, concentration, or spatial distribution of heteroatoms and size-controlled vacancies within a solid, along with materials processing approaches, such as three-dimensional structure assembly and surface metal complexation methodologies, has demonstrated its potential to tackle the challenges triggered by energy conversion concerns in direct alcohol fuel cells. With continuous progress on the knowledge The author acknowledges the Brazilian National Council for Scientific and Technological Development (CNPq) for research funding.


**3. Methods**

*Assistive and Rehabilitation Engineering*

loop. Current technology, however, is not able to address the complex issue of hand function, which involves overlapping neuro-physio strategies and multiple degrees of freedom. At most, simple movements may be possible [32] which has been shown to not adequately impact function for the highly heterogeneous stroke affected population. Gross movements can be expected to improve with very high number of repetitions, thus enabling the brain to rewire itself in a limited way. However, there is poor evidence that such gross movement practice translates significantly into function. Therefore, the modification to the above model is proposed, incorporating the feedforward and feedback elements modeled in **Figure 2** as a form of

The augmented feedback may be delivered visually via a muscle-brain-computer interface. The feedforward in the form of appropriate audio-visual inputs, which lead the human to attempt a series of desired actions through imitation, is known to facilitate recovery [33]. Moreover, there is evidence of perception transferring to action and more importantly, from action to perception [34]. The augmented feedback is expected to drive motor intention and exploration while the feedforward is expected to prime the brain for motor actuation and goal directed learning through imitation. From a functional improvement perspective, the augmented feedback

augmentation to help overcome the deficits through the learning route.

may be customized for a person using time-locked parameters as follows:

The brain and the body are inseparably linked, and both contribute significantly for neuroplasticity to occur and health parameters to improve [35]. Based on this understanding of how human learning may be applied practically in the context of post-stroke rehabilitation, this study was conceived with the following

1.When EEG and EMG signals during activity are brought together in a timesynchronized manner for real-time feedback along with an audio-visual feedforward for imitation, it provides an opportunity for the patient to work with sensory, exploratory and goal-directed learning toward functional

2.Displaying quantified, relative brain and muscle feedback in real-time while training activation and relaxation simultaneously during movement or while attempting to manipulate objects, will enhance the conditions for incremental associative learning of overlapping brain and muscle strategies to occur [36]. Under such conditions, subjects may potentially achieve systemic gains in functional performance, even though they may have tried existing rehabilita-

This paper describes a bio-mechatronics approach to understanding where re-learning is misled or failing and uses a "feedforward-feedback" modality to help chronic stroke subjects train gross movements (as measured by Fugyl Meyer Upper Extremity Motor Assessment scale) and functional, timed-task capabilities (as measured by Action Research Arm Test). The SynPhNe system employs learning and training principles similar to that which babies seem to use in the design of its user interface, to leverage the mechanism of "self-regulation" or "self-correction." The study explores to what extent such real-time "self-correction" alone, in the absence of any form of external stimulation or robotic assistance, impacts the

1.EMG agonist-antagonist balance (muscle strategy).

2.EEG relaxation and attention states (brain strategy).

tion methods and only partially succeeded.

**120**

assumptions:

rehabilitation goals.


**Neurocognitive Effects of Primary Brain Tumors**

Mohammad Abu-Hegazy and Hend Ahmed El-Hadaad

Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/62924

#### **Abstract**

[68] Chamberlain MC, Tredway TL. Adult primary intradural spinal cord tumors: a review.

[69] Deletis V, Sala F. Intraoperative neurophysiological monitoring of the spinal cord during spinal cord and spine surgery: a review focus on the corticospinal tracts. Clin

[70] Sala F, Bricolo A, Faccioli F, Lanteri P, Gerosa M. Surgery for intramedullary spinal cord tumors: the role of intraoperative (neurophysiological) monitoring. Eur Spine J.

[71] Kothbauer K, Deletis V, Epstein FJ. Intraoperative spinal cord monitoring for intrame‐

[72] de Haan P, Kalkman CJ, Ubags LH, Jacobs MJ, Drummond JC. A comparison of the sensitivity of epidural and myogenic transcranial motor-evoked responses in the detection of acute spinal cord ischemia in the rabbit. Anesth Analg. 1996; 83: 1022–1027.

[73] Hyun SJ, Rhim SC. Combined motor and somatosensory evoked potential monitor‐ ing for intramedullary spinal cord tumor surgery: correlation of clinical and neuro‐ physiological data in 17 consecutive procedures. Br J Neurosurg. 2009; 23(4): 393–400.

[74] Skinner SA, Nagib M, Bergman TA, Maxwell RE, Msangi G. The initial use of freerunning electromyography to detect early motor tract injury during resection of

intramedullary spinal cord lesions. Neurosurgery. 2005; 56 (2 Suppl): 299–314.

[75] Skinner SA, Transfeldt EE. Electromyography in the detection of mechanically induced spinal motor tract injury: observations in diverse porcine models. J Neurosurg Spine.

[76] Skinner SA, Hsu B, Transfeldt EE, Mehbod AA, Rippe DM, Wu C, et al. Spinal cord injury from electrocautery: observations in a porcine model using electromyography

and motor evoked potentials. J Clin Monit Comput. 2013; 27(2): 195–201.

dullary surgery: an essential adjunct. Pediatr Neurosurg. 1997: 247–254.

Curr Neurol Neurosci Rep. 2011; 11(3): 320–328.

Neurophysiol. 2008; 119(2): 248–264.

2007; 16 Suppl 2: S130–9.

240 Neurooncology - Newer Developments

2009; 11(3): 369–374.

Cognitive impairment, a common finding with the brain tumors, may result from the tumor itself or the treatment used: surgery, chemotherapy, or radiotherapy. Surgery for brain tumors improves the cognitive function due to reduction of compression as in case of removal of noninvasive tumors. Stability of cognitive function also was observed after tumor resection, such as tumors of third ventricle. Postoperative cognitive worsening was observed. Postoperative worsening of executive functions may correlate to volume of the operated area. Cognitive deficits may follow radiotherapy by several months to many years. These deficits may be due to vascular injury, local radionecrosis, and cerebral atrophy. This usually involves multiple domains, including memory, attention, executive function, and intelligence. The irradiated volume of brain tissue has great impact on cognition. Intensity-modulated radiotherapy (IMRT) and proton beam therapy result in greater sparing of healthy brain tissue and allow for a more-targeted delivery of radiation and smaller penetration of tissue beyond the tumor consequently reduce the risk of cognitive deficit after radiotherapy. Chemotherapy treatment in brain tumor seems to have a role in cognitive dysfunction deficits. The toxicity of chemotherapy increased when was given during or after radiotherapy. Chemotherapeutic agents, such as BCNU, CDDP, cytosine arabinoside, and intrathecal or intravenous methotrexate, have toxic effect to the CNS. Glioblastoma patients undergoing radiotherapy with concomitant and adjuvant temozolomide treatment do not develop cognitive deterioration. Patients with brain tumors face the challenge of cognitive impairment due to the tumor itself or treatments. Cognitive deficits in processing speed, memory, attention, and executive functions interfere with patients' daily life activities. Cognitive rehabilitation program has proven to be effective in patients with primary brain tumors. Cognitive impairments have a large impact on self-care, social and professional functioning, and consequently on quality of life. Preventing these late effects is a challenge for the medical team, psychologists, and rehabilitation specialists. Prevention depends in part on being able to predict those at greatest risk. Advances in neurosurgery, chemotherapy, and radiotherapy techniques are helping to a great extent, but may not be totally successful at preventing these late effects.

© 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


*2.2. Plant Location and Capacity*

In this study, Iran is selected as a place for the biorefinery, where fossil fuel resources and methanol are widely available. However, the area has water shortages and is dealing with severe environmental impacts. Therefore, castor, as a low water demanding plant that can be cultivated in wastewater, is used. Considering the total world production of castor [9,27], a plant with the capacity of handling 120,000 tons of castor plant per year (based on the dry weight) is used.


**9. The physical and chemical properties of biodiesel produced from algal cell**

Analysis of the produced biodiesel from the promising alga *Dictyochloropsis splendida* (Table 5). showed that the unsaturated fatty acids percentage was increased in alga cultivated in nitrogen free media (0.0g/l N) two times more than normal conditions (13.67, 4.81% respectively). However, the composition of fatty acids was different in these algae depending on its growth condition as showed in table 3. These results were in agreements with those reported by Wood (1974) relative to *Chlorophycean* species. Furthermore Ramos *et al*. (2009) reported that monounsaturated, polyunsaturated and saturated methyl esters were built in order to predict the critical parameters of European standard for any biodiesel, composition. The extent of unsaturation of microalgae oil and its content of fatty acids with more than four double bonds can be reduced easily by partial catalytic hydrogenation of the oil (Jang *et al*., 2005, Dijkstra, 2006). Concerning the fatty acids contents of the produced biodiesel from microalgae, Chisti (2007) reported in his review that, microalgal oils differ from vegetable oils in being quite rich in polyunsaturated fatty acids with four or more double bands (Belarbi *et al*., 2000) as eicosapentanoic acid (C20:5n-3) and docosahexaenoic acid (C22:6n-3) which occurred commonly in algal oils. The author added that, fatty acids and fatty acid methyl esters with four and more double bands are susceptible to oxidation during storage and this reduces their acceptability for use in biodiesel especially for vehicle use (European standard EN 14214 limits to 12%) while no such limitation exists for biodiesel intended for use as healing oil. In addition to the content of unsaturated fatty acids in the biodiesel also its iodine value (represented total unsaturation) must be taken in consideration (not exceeded 120 g iodine/100g biodiesel according to the European standard.

410 2.7 21c

49 0.3 25c

Table 4. Comparison of estimated biodiesel production efficiencies from vascular plants and

**9. The physical and chemical properties of biodiesel produced from algal cell**  Analysis of the produced biodiesel from the promising alga *Dictyochloropsis splendida* (Table 5). showed that the unsaturated fatty acids percentage was increased in alga cultivated in nitrogen free media (0.0g/l N) two times more than normal conditions (13.67, 4.81% respectively). However, the composition of fatty acids was different in these algae depending on its growth condition as showed in table 3. These results were in agreements with those reported by Wood (1974) relative to *Chlorophycean* species. Furthermore Ramos *et al*. (2009) reported that monounsaturated, polyunsaturated and saturated methyl esters were built in order to predict the critical parameters of European standard for any biodiesel, composition. The extent of unsaturation of microalgae oil and its content of fatty acids with more than four double bonds can be reduced easily by partial catalytic hydrogenation of the oil (Jang *et al*., 2005, Dijkstra, 2006). Concerning the fatty acids contents of the produced biodiesel from microalgae, Chisti (2007) reported in his review that, microalgal oils differ from vegetable oils in being quite rich in polyunsaturated fatty acids with four or more double bands (Belarbi *et al*., 2000) as eicosapentanoic acid (C20:5n-3) and docosahexaenoic acid (C22:6n-3) which occurred commonly in algal oils. The author added that, fatty acids and fatty acid methyl esters with four and more double bands are susceptible to oxidation during storage and this reduces their acceptability for use in biodiesel especially for vehicle use (European standard EN 14214 limits to 12%) while no such limitation exists for biodiesel intended for use as healing oil. In addition to the content of unsaturated fatty acids in the biodiesel also its iodine value (represented total unsaturation) must be taken in consideration (not exceeded 120 g iodine/100g biodiesel according to the European

**Area required as a percent of total global land**

**Area required as a percent of total arable global land**

**Area needed to meet global oil demand (106 hectans)**

cAssuring that microalgal ponds and bioreactors are located on non-arable land

**Cotton** 15000 101 757 **Soybean** 10900 73 552 **Mastard seed** 8500 57 430 **Sunflower** 5100 34 258 **Rapeseed/Canola** 4100 27 207 **Jatropha** 2600 17 130b **Oil palm** 820 5.5 41

**Biodiesel feedstock**

**Microalgae (10 g/m3/day, 30%TAG)**

**Microalgae (50 g/m3/day, 50%TAG)**

microalgae

standard.

b Jatropha is mainly grown on marginal land

Table 5. Analysis of fatty acids of the obtained biodiesel from the promising green microalgae *Dictyochloropsis* sp

#### **10. Enhancement the biodiesel production from algae**

Lipid productivity, the mass of lipid that can be produced per day, is dependent upon plant biomass production as well as the lipid content of this biomass. Algal biodiesel production will therefore be limited not only by the standing crop of microalgae, but also by its lipid content, which can vary from <1% to >50% dry weight [Shifrin, N.S. and Chisholm, 1980]. Given that a strong and predictable response of microalgal biomass to phosphorus enrichment has consistently been exhibited by freshwater ecosystems worldwide (Box 2), it can be expected that the volumetric lipid content (in mg L\_1) of water contained in algal bioreactors should also in general increase with an increase in the total phosphorus content of the system, as has been reported for lakes by Berglund *et al*. [Berglund, 2001]. However, both the quantity and the quality of lipids produced will vary with the identity of the algal species that are present in the water, as well as with site-specific growth conditions. This variability probably reflects modifications in the properties of cellular membranes, and alterations in the relative rates of production and utilization of storage lipids [Roessler, 1990]. In the presence of moderate temperatures and sufficient light, many dozens of studies during the past several decades have revealed that algal lipid content is particularly sensitive to conditions of nutrient limitation . For example, silicon-starved diatoms can contain almost 90% more lipids than silicon-sufficient cells [Shifrin, N.S. and Chisholm, 1980]. However, silicon will be a growth-limiting nutrient only for the limited subset of microalgal species that have an absolute requirement of this element for their cellular growth. A stronger stimulation of lipid production occurs in response to conditions of nitrogen limitation, which potentially can occur in all known microalgae. Nitrogen-starved cells can contain as much as four times the lipid content of Nsufficient cells [Shifrin, N.S. and Chisholm, 1980], and maximizing the lipid production of pond bioreactors should

Algal Biomass and Biodiesel Production 125

biodiesel and sediment (glycerin and pigments) percentages. Hexane/ ether (1:1, v/v) extraction solvent system resulted in low lipid recoveries (2.3-3.5% dry weight) while; chloroform/methanol (2: 1, v/v) extraction solvent system was proved to be more efficient for lipid and biodiesel extraction (2.5 – 12.5% dry weight) depending on algae species (Table 7). The green microalga *Dictyochloropsis splendida* extract produced the highest lipid and biodiesel yield (12.5 and 8.75% respectively) followed by the cyanobacterium *Spirulina maxima* (9.2 and 7.5 % respectively). On the other hand, the macroalga (red, brown and green) produced the lowest biodieselyield. The fatty acids of *Dictyochloropsis splendida*  Geitler biodiesel were determined using gas liquid chromatography. Lipids, biodiesel and glycerol production of *Dictyochloropsis splendida* Geitler (the promising alga) were markedly enhanced by either increasing salt concentration or by nitrogen deficiency (Table 8) with maximum production of (26.8, 18.9 and 7.9 % respectively) at nitrogen starvation condition.

Table 7. Total lipid, biodiesel, sediment percentage and biodiesel color of eight algal species Natural biotic communities in outdoor bioreactors require the external provision of potentially growth-limiting resources (e.g. light, carbon dioxide and the essential mineral nutrients N and P). These resources act as ''bottom-up'' regulators of the potential microalgal biomass that can be produced. Once harvested, the cellular lipids in this microalgal biomass can be extracted and processed to create biodiesel fuels. The lipid content of microalgal biomass is not constant, however, and can be influenced by many factors, including nitrogen:phosphorus supply ratios, light, CO2 and the hydraulic residence time of the bioreactor. Moreover, natural assemblages of microalgae are taxonomically diverse: some species are small and can easily be consumed by herbivorous zooplankton. Undesirable grazing losses of edible microalgae (and their cellular lipids) to large-bodied zooplankton can be reduced by adding zooplanktivorous fish, which can greatly restrict large-bodied zooplankton growth via sizeselective predation (''top-down'' regulation).

(Afify *et al*., 2010)

therefore depend on their operators' ability to reliably and consistently induce N-limitation in the resident algal cells. Resource-ratio theory and the principles of ecological stoichiometry, provide additional new insights into the control of algal biomass and lipid production in pond bioreactors. the nutrient limitation status of microalgae can be directly controlled by regulating the ratio of nitrogen and phosphorus (N:P) supplied in the incoming nutrient feed: nitrogen limitation occurs at N:P supply ratios that lie below the optimal N:P ratio for microalgal growth, whereas phosphorus limitation occurs at ratios that exceed this ratio. A transition between N- and P-limitation of phytoplankton growth typically occurs in the range of N:P supply ratios between ca. 20:1 to ca. 50:1 by moles . Such shifts between N- and P-limitation have extremely important implications for algal biofuel production because diverse species of microalgae grown under nitrogen-limited conditions (i.e. low N:P supply ratios) can exhibit as much as three times the lipid content of cells grown under conditions of phosphorus limitation (high N:P supply ratios) . Both the total phosphorus concentration as well as the total nitrogen concentration in the nutrient feeds to pond bioreactors should therefore impact algal biodiesel production, because the N:P ratio of incoming nutrients will strongly influence algal biomass production as well as the cellular lipid content. Given the inverse relationship observed between N:P and cellular lipids , and the positive, hyperbolic relationship observed between N:P and microalgal biomass , we conclude that optimal lipid yields (in terms of mass of lipid produced per unit bioreactor volume per day) should occur at intermediate values of the N:P supply ratio. From the strong apparent interactions between the effects of nitrogen and carbon dioxide availability on microalgal lipids, we also conclude that the effects of N:P supply ratios on volumetric lipid production might be even greater if the bioreactors are simultaneously provided with supplemental CO2 (cf. Figure 2).

Table 6. Comparison between lipid percentage (%) produced by eight algal species using two different extraction system.

Eight algal species (4 *Rhodo*, 1 *chloro* and 1 *phaeophycean* macroalgae, 1 *cyanobacterium* and 1 green microalga) were used for the production of biodiesel using two extraction solvent systems (Hexane/ether (1:1, v/v)) and (Chloroform/ methanol (2:1, v/v)) Table 6. Biochemical evaluations of algal species were carried out by estimating biomass, lipid,

therefore depend on their operators' ability to reliably and consistently induce N-limitation in the resident algal cells. Resource-ratio theory and the principles of ecological stoichiometry, provide additional new insights into the control of algal biomass and lipid production in pond bioreactors. the nutrient limitation status of microalgae can be directly controlled by regulating the ratio of nitrogen and phosphorus (N:P) supplied in the incoming nutrient feed: nitrogen limitation occurs at N:P supply ratios that lie below the optimal N:P ratio for microalgal growth, whereas phosphorus limitation occurs at ratios that exceed this ratio. A transition between N- and P-limitation of phytoplankton growth typically occurs in the range of N:P supply ratios between ca. 20:1 to ca. 50:1 by moles . Such shifts between N- and P-limitation have extremely important implications for algal biofuel production because diverse species of microalgae grown under nitrogen-limited conditions (i.e. low N:P supply ratios) can exhibit as much as three times the lipid content of cells grown under conditions of phosphorus limitation (high N:P supply ratios) . Both the total phosphorus concentration as well as the total nitrogen concentration in the nutrient feeds to pond bioreactors should therefore impact algal biodiesel production, because the N:P ratio of incoming nutrients will strongly influence algal biomass production as well as the cellular lipid content. Given the inverse relationship observed between N:P and cellular lipids , and the positive, hyperbolic relationship observed between N:P and microalgal biomass , we conclude that optimal lipid yields (in terms of mass of lipid produced per unit bioreactor volume per day) should occur at intermediate values of the N:P supply ratio. From the strong apparent interactions between the effects of nitrogen and carbon dioxide availability on microalgal lipids, we also conclude that the effects of N:P supply ratios on volumetric lipid production might be even greater if the bioreactors are simultaneously

Table 6. Comparison between lipid percentage (%) produced by eight algal species using

Eight algal species (4 *Rhodo*, 1 *chloro* and 1 *phaeophycean* macroalgae, 1 *cyanobacterium* and 1 green microalga) were used for the production of biodiesel using two extraction solvent systems (Hexane/ether (1:1, v/v)) and (Chloroform/ methanol (2:1, v/v)) Table 6. Biochemical evaluations of algal species were carried out by estimating biomass, lipid,

provided with supplemental CO2 (cf. Figure 2).

two different extraction system.

biodiesel and sediment (glycerin and pigments) percentages. Hexane/ ether (1:1, v/v) extraction solvent system resulted in low lipid recoveries (2.3-3.5% dry weight) while; chloroform/methanol (2: 1, v/v) extraction solvent system was proved to be more efficient for lipid and biodiesel extraction (2.5 – 12.5% dry weight) depending on algae species (Table 7). The green microalga *Dictyochloropsis splendida* extract produced the highest lipid and biodiesel yield (12.5 and 8.75% respectively) followed by the cyanobacterium *Spirulina maxima* (9.2 and 7.5 % respectively). On the other hand, the macroalga (red, brown and green) produced the lowest biodieselyield. The fatty acids of *Dictyochloropsis splendida*  Geitler biodiesel were determined using gas liquid chromatography. Lipids, biodiesel and glycerol production of *Dictyochloropsis splendida* Geitler (the promising alga) were markedly enhanced by either increasing salt concentration or by nitrogen deficiency (Table 8) with maximum production of (26.8, 18.9 and 7.9 % respectively) at nitrogen starvation condition. (Afify *et al*., 2010)

Table 7. Total lipid, biodiesel, sediment percentage and biodiesel color of eight algal species

Natural biotic communities in outdoor bioreactors require the external provision of potentially growth-limiting resources (e.g. light, carbon dioxide and the essential mineral nutrients N and P). These resources act as ''bottom-up'' regulators of the potential microalgal biomass that can be produced. Once harvested, the cellular lipids in this microalgal biomass can be extracted and processed to create biodiesel fuels. The lipid content of microalgal biomass is not constant, however, and can be influenced by many factors, including nitrogen:phosphorus supply ratios, light, CO2 and the hydraulic residence time of the bioreactor. Moreover, natural assemblages of microalgae are taxonomically diverse: some species are small and can easily be consumed by herbivorous zooplankton. Undesirable grazing losses of edible microalgae (and their cellular lipids) to large-bodied zooplankton can be reduced by adding zooplanktivorous fish, which can greatly restrict large-bodied zooplankton growth via sizeselective predation (''top-down'' regulation).

Algal Biomass and Biodiesel Production 127

Governorate, Egypt was used after filtered using glass microfiber filter to remove large particles and indigenous bacteria for the experiment and the chemical and physical parameters were analysis as reported by APHA (1998) Table (2). The supplementation of NaNO3, K2HPO4 and FeSO4.7H2O in amounts equal to those of the standard BG11, Bold and Zarrouk were used as basal media. The algal strains were grown in 500 ml Erlenmeyer flasks containing 200 ml of 100% effluent supplemented with basal nutrients and 100% effluent without basal nutrients with/without sterilization and the synthetic media (BG11, Bold and Zarrouk) were used as control. Two per cent algal inoculums were added to each flask. The experiment was conducted in triplicates and cultures were incubated at 25 ºC ±1ºC, under continuous shaking (150 rpm) and illumination (2000 lux) for 15 days. This work aimed to evaluate the laboratory cultivation of nine algal strains belonging to Nostocales and Chlorellales in secondary treated municipal domestic wastewater for

**Color pH Glycerin +**

7.5±0.50 Yellowish

**pigments**

biomass and biodiesel production as shown in Table (9 and 10).

**content Algal species Total lipids**

*muscorum*

*aquae*

*vulgaris*

*platensis*

*oryzae*

*humifusum*

according to Duncan's multiple range tests.

sterilization T4: waste water+ nutrients without sterilization

from different microalgae species cultivated in different waste water

**Biodiesel**

16.80±3.62 12.52±1.74 4.28±1.74 Brown 7.4±0.33 *Nostoc*

5.50±0.58 4.00±0.41 1.50±0.41 Red 6.9±0.95 *Anabaena flous*

12.50±1.20 8.8±0.16 3.70±0.16 Green 8.1±1.0 *Chlorella*

*Oscillatoria sp* 8.00±0.58 4.30±0.32 3.70±0.32 Yellow 7.5±0.85

10.0±0.11 7.80±0.17 2.20±0.17 Light green 8.0±0.32 *Spirulina*

7.40±0.90 4.50±0.10 2.90±0.10 Orange 7.3±0.96 *Anabaena*

*Wollea sp* 6.30±1.31 3.90±0.60 2.40±0.60 Yellow 7.8±0.35

*Phormedium sp* 12.20±1.66 10.10±1.50 2.10±1.50 Dark brown 7.1±0.0

Each value is presented as mean of triplet treatments, LSD: Least different significantly at P ≤ 0.05

T1: waste water without treatment; T2: waste water after sterilization; T3: waste water+ nutrients with

Table 9. Total lipids, biodiesel, glycerine+pigments percentage and color, pH of biodiesel

LSD 0.159 0.151 0.151 1.659

brown 14.80±2.40 10.20±1.30 4.6±1.30 *Nostoc*

Table 8. Total lipid, biodiesel, sediment percentage and biodiesel color of Dictyochloropsis sp cultivated under stress


**2.2 Characterization and stack effective** *R* **genes in hybrid rice**

Following the clone of resistant genes and the development of related functional molecular makers, marker screening has been widely applied on resistance identification and innovation of parents of hybrid rice. Up to now, molecular makers of blast resistance genes, *Pi2*, *Pi9*, *Pi1*, *Pib*, *Pita*, *Pid2*, *Pikh*, *Pigm*, and *Pish*, have been developed and used on detecting the related *R* genes [17–25]. In Jiangsu, a total of 544 rice materials were assessed for blast resistance and resistance genes distribution by inoculation and marker screening; results showed that 968, Xiushui 134, Jia 58, Jindao 263, Huaidao 20, Yandao 10, and Gumei 4 contained the majority of resistance genes; and *Pi5*, *Pita*, *Pi9*, and *Pib* exhibited high resistance to six major blast races [14]. In South China, with functional marker of *Pi1*, *Pik-p*, *Pikh*, *Pi2*, *Pi9*,

**29**

**Table 2.**

**Figure 1.**

*The Utilization of Rice Blast Resistance Genes in Hybrid Rice Breeding in China*

make breeders directly utilize related resistance resources on purpose.

*Piz-t*, *Pita*, and *Pii*, 328 hybrid rice combinations were screened, in which *Pita* and *Pii* were found in high frequency, but *Pi2* and *Pi1* displayed highly effective resistance contribution to local rice [26]. In Sichuan Province, with molecular markers closely linked to *Pi-9*, *Pi-2*, *Pi-kh*, and *Pi-km*, general rice parents of hybrid rice were analyzed and selected for the resistance resources [27]. The *R* gene screening

The hybridization, backcross, and marker-assisted selection (MAS) were the general method for the introduction of *R* genes into the restorer line, maintainer line, and sterile line of hybrid rice. MAS conduced to selectively breeding based on the genotype and accelerate the breeding course [28]. The procedure for MAS was shown in **Figure 1**. As we know, *R* genes, such as *Pi9*, *Pi2*, *Pi1*, and *Pigm*, have been reported to show relatively high resistance in different districts in China [29–31]. Hence, these *R* genes were often used for improvement of rice blast

**R genes used Variety improved Variety type Reference** *Pi9* Yandao 6 Hao General cultivar [32] *Pi25* Xiangwanxian 13 General cultivar [33] *Pi1, Pi2* and *Pi33* Jin 23B Maintainer line [34] *Pi1* and *Pi2* Rongfeng B Maintainer line [35] *Pid*(t)*, Pib* and *Pita* G46B Maintainer line [36] *Pi9* R599 Maintainer line [37] *Pi9* R288 Maintainer line [38] *Pi9* Shuhui527, Minghui 86, and Minhui 3301 Restorer line [39] *Pigm(*t) Chunhui 350 Restorer line [40] *Pi9* and *Pi49* Chuang 5S Sterile line [41] *Pi25* Zhenda A Sterile line [42] *Pi47* and *Pi48* C815S Sterile line [43] *Pi9* 03S Sterile line [44] *Pi1* GD-8S Sterile line [45] *Pi1* and *Pi2* Peiai64S Sterile line [46]

*DOI: http://dx.doi.org/10.5772/intechopen.83617*

*The breeding course with marker-assisted selection and backcross.*

*The improved rice varieties of different types with MAS technique.*

## **Table 1.**

*Selected rice varieties with different resistant resources to blast in China.*

*The Utilization of Rice Blast Resistance Genes in Hybrid Rice Breeding in China DOI: http://dx.doi.org/10.5772/intechopen.83617*

*Piz-t*, *Pita*, and *Pii*, 328 hybrid rice combinations were screened, in which *Pita* and *Pii* were found in high frequency, but *Pi2* and *Pi1* displayed highly effective resistance contribution to local rice [26]. In Sichuan Province, with molecular markers closely linked to *Pi-9*, *Pi-2*, *Pi-kh*, and *Pi-km*, general rice parents of hybrid rice were analyzed and selected for the resistance resources [27]. The *R* gene screening make breeders directly utilize related resistance resources on purpose.

The hybridization, backcross, and marker-assisted selection (MAS) were the general method for the introduction of *R* genes into the restorer line, maintainer line, and sterile line of hybrid rice. MAS conduced to selectively breeding based on the genotype and accelerate the breeding course [28]. The procedure for MAS was shown in **Figure 1**. As we know, *R* genes, such as *Pi9*, *Pi2*, *Pi1*, and *Pigm*, have been reported to show relatively high resistance in different districts in China [29–31]. Hence, these *R* genes were often used for improvement of rice blast

#### **Figure 1.**

*Protecting Rice Grains in the Post-Genomic Era*

resistance breeding.

**2. The utilization of rice blast resistance genes**

The resistance level of parents is directly related with the resistance performance

Following the clone of resistant genes and the development of related functional molecular makers, marker screening has been widely applied on resistance identification and innovation of parents of hybrid rice. Up to now, molecular makers of blast resistance genes, *Pi2*, *Pi9*, *Pi1*, *Pib*, *Pita*, *Pid2*, *Pikh*, *Pigm*, and *Pish*, have been developed and used on detecting the related *R* genes [17–25]. In Jiangsu, a total of 544 rice materials were assessed for blast resistance and resistance genes distribution by inoculation and marker screening; results showed that 968, Xiushui 134, Jia 58, Jindao 263, Huaidao 20, Yandao 10, and Gumei 4 contained the majority of resistance genes; and *Pi5*, *Pita*, *Pi9*, and *Pib* exhibited high resistance to six major blast races [14]. In South China, with functional marker of *Pi1*, *Pik-p*, *Pikh*, *Pi2*, *Pi9*,

**Province Resistance resource Reference**

Dongnong 415, Songjing 9, Longdao 12, Hejiang 23, and Wuyoudao 3

610, Hanyou 983, Lvyinzhan, Bingyou C278, Yuenongsimiao, and

Guangdong Sanhuangzhan 2 Hao, Qingliuai 1 Hao, Jingxian 89, IR36, and 28 Zhan [9] Hunan Xiangzao 143, Fengyuanyou 299, Jinyou 207, Liangyou 222, Quanfengyou

[6, 7]

[8]

[10, 11]

[12, 13]

[14, 15]

[16]

Sichuan IR99–35, Miyang 46, IR 1544, Tetep, Gumei 2, 6326, Suhui162, and Suhui

Hubei Zhenke, Jinlong 1, Fanyu 1, Ningwan 1, Sanqizao, Nanjing 15, Aiyinnuo,

Fujian Yixiangyou 673, Dyou 15, Gangyou 148, Guyou 527, Jiafuzan, and Teyou

Jiangsu Longjing 968, Xiushui 134, Jia 58, Jindao 263, Huaidao 20, Yandao 10, and

Jinzao 47, Yunjin 23, and Quanzhen 10

Heilongjiang Suijing 12, Mudanjiang 26, Longdun 105, Longjing 20, Longjing 31,

527

627

Gumei 4

*Selected rice varieties with different resistant resources to blast in China.*

Zhuoliangyou 249

of hybrid rice. The resistance evaluation for breeding lines is a very important prerequisite work for resistance breeding of hybrid rice. For traditional breeders, field nursery or artificial inoculation with blast isolates in greenhouse was normally used for resistance identification. Amounts of rice lines with middle or high resistance have been identified in different provinces with diverse ecology. The detailed information was listed in **Table 1**. These identified rice materials provided rich selection as parents or resistant resource for hybrid rice breeding. As we know, genetic mechanism of rice blast resistance followed the gene for gene interaction. It was unclear about background and resistant genes in these materials, and the presence of one *R* gene masked another *R* gene; and also, the stationary field nursery only can stand for limited ecological districts. Hence, blast evaluation cannot identify any particular resistance gene, and it will lead to huge uncertainty in resistance of later generations in breeding. Phenotype identification cooperated with precise analysis of resistant genes will more effectively serve for hybrid rice

**2.2 Characterization and stack effective** *R* **genes in hybrid rice**

**2.1 Identification of the resistant rice parents**

**28**

**Table 1.**

*The breeding course with marker-assisted selection and backcross.*

### **Table 2.**

*The improved rice varieties of different types with MAS technique.*

resistance (**Table 2**). Recently, a new class resistance gene *Ptr* just cloned encoded an atypical protein and conferred broad-spectrum disease resistance and will provide diverse selection for resistance improvement [47]. To breeding rice cultivars with durable blast resistance, stacking several resistance genes still was the most effective method. To stack resistance gene purposefully, spectrum of each resistance gene must be determined, and also, the identification of differential blast races/isolates that distinguished each resistance gene in different districts was critical for ensuring the effectiveness of resistance gene stacking [48].

## **3. Conclusions**

With identification in the rice blast field nursery or functional marker detecting of major *R* genes, the amount of blast resistance resources was identified and provides diverse selections for hybrid rice resistance breeding. However, the recent finding showed that *Pita* required *Ptr* to function revealed that part of single *R* gene may be not functional as we thought originally [47], and further function analysis of more *R* genes may be necessary.

For conventional rice breeding, all blast *R* genes must be stacked into breeding lines to be effective, whereas hybrid rice can stack blast *R* genes into two parents. For hybrid rice breeding, blast resistance was only a part of index, and other agronomic traits also need to be considered. Lines of Chuang 5 S stacked *Pi9* and *Pi49* have been found obvious differences on plant height, spike length, spike number and stigma exertion rate with the receptor, even though the blast resistance has improved [41]. Hence, blast resistance breeding for hybrid rice was a synthetic work that contained resistance innovation and excellent agronomic trait selection.

In this chapter, it introduced the progress on identification of resistance resources and the utilization of blast resistance genes. Traditional cross technique, combined with MAS, has been used to transfer different major *R* genes into parent's lines to improve the resistance of hybrid rice and achieved remarkable results. Following the improvement of blast resistance of the authorized varieties, it gratefully contributed to decrease the damage of rice blast disease and played important function on protection of rice production safety in China.

### **Acknowledgements**

This work was supported by Hunan Provincial Science and Technology Department (CN) (2018NK1020), Hunan Academy of Agricultural Science and Technology Innovation Project (2018ZD01-7), Hunan Academy of Agricultural Science and Technology Innovation Project (2017JC03), and Hunan Provincial Natural Science Fund Youth Fund (2018JJ3379).

**31**

**Author details**

provided the original work is properly cited.

Junjie Xing\*, Huafeng Deng and Longping Yuan

Hunan Hybrid Rice Research Center, Changsha, Hunan, China

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

*The Utilization of Rice Blast Resistance Genes in Hybrid Rice Breeding in China*

*DOI: http://dx.doi.org/10.5772/intechopen.83617*

### **Conflict of interest**

The authors declared that there was no conflict of interest.

*The Utilization of Rice Blast Resistance Genes in Hybrid Rice Breeding in China DOI: http://dx.doi.org/10.5772/intechopen.83617*


**Chapter 21**

*Sustainability in Urban Planning and Design*

neighborhood walkability with street data based on geographic information systems. Transportation Research Record: Journal of the transportation research board. 1982;**2006**:48-56

design. Journal of Urban Design.

[49] Moudon A, Hess P, Snyder M, Stanilov K. Effects of site design on pedestrian travel in mixed-use, medium-density environments. Transportation Research Record: Journal of the Transportation Research

[50] Remali AM, Porta S, Romice O, Abudib H, Vaughan L. Street quality, street life, street centrality. Suburban Urbanities: Suburbs and the life of the high street: UCL Press; 2015.

[51] Gehl J. Public Spaces and Public Life in Perth: Report for the Government of Western Australia and the City of Perth. Curtin University Research Publications; 1994. Available from: http://hdl.handle.

net/20.500.11937/15502

[52] Hershberger RG. Architecture and meaning. Journal of Aesthetic Education. 1 Oct 1970;**4**(4):37-55

[53] Ewing R, Handy S, Brownson RC, Clemente O, Winston E. Identifying and measuring urban design qualities

related to walkability. Journal of Physical Activity and Health.

2006;**3**(s1):S223-S240

1998;**3**(1):93-116

Board. 1997;**1578**:48-55

pp. 104-129

[39] Cerin E, Leslie E, du Toit L, Owen N, Frank LD. Destinations that matter: Associations with walking for transport. Health and Place. 2007;**13**(3):713-724

[40] Lee C, Moudon AV. Correlates of walking for transportation or recreation purposes. Journal of Physical Activity and Health. 2006;**3**(s1):S77-S98

[41] Lee C, Moudon AV. The 3Ds+ R: Quantifying land use and urban form correlates of walking. Transportation Research Part D: Transport and Environment. 2006;**11**(3):204-215

[42] Randall TA, Baetz BW. Evaluating pedestrian connectivity for suburban sustainability. Journal of Urban Planning and Development. 2001;**127**(1):1-15

[43] Moudon AV, Hess PM. Suburban

American Planning Association.

[44] Canter D, Tagg SK. Distance estimation in cities. Environment and

[45] Watts DJ, Strogatz SH. Collective dynamics of 'small-world'networks. Nature. 1998;**393**(6684):440-452

[46] Van der Westhuizen DL. Concepts of space and place neighborhood access, pedestrian movement, and physical activity in Detroit: Implications for urban design and research. University of

[47] Carmona M. Public Places, Urban Spaces: The Dimensions of Urban Design.

Burlington: Elsevier; 2010

[48] Montgomery J. Making a city: Urbanity, vitality and urban

clusters. Journal of the

Behavior. 1975;**7**(1):59-80

2000;**66**(3):243-264

Michigan; 2010

**386**


*Article* **Effect of Depth-Induced Breaking on Wind Wave Simulations in Shallow Nearshore Waters off Northern Taiwan during the Passage of Two Super Typhoons**

**Shih-Chun Hsiao 1, Han-Lun Wu 1, Wei-Bo Chen 2,\*, Wen-Dar Guo 2, Chih-Hsin Chang 2 and Wen-Ray Su 2**

1 Department of Hydraulic and Ocean Engineering, National Cheng Kung University, TainanCity70101,Taiwan;schsiao<EMAIL_ADDRESS>
 (H.-L.W.) 2 National Science and Technology Center for Disaster Reduction, New Taipei City 23143, Taiwan; (W.-D.G.);(C.-H.C.);wrsu@ncdr.nat.gov.tw(W.-R.S.)

<EMAIL_ADDRESS><EMAIL_ADDRESS>**\***Correspondence:wbchen@ncdr.nat.gov.tw;Tel.:+886-2-8195-8611

**Abstract:** Super Typhoons Maria (2018) and Lekima (2019) were adopted for this case study, although they only passed the northern offshore waters of Taiwan without making landfall. A direct modification technique was employed to create the atmospheric conditions for a wave-circulation model to hindcast large typhoon-driven waves. The radius of the modified scale (*Rtrs*) for a hybrid typhoon wind plays an important role in the significant wave height (SWH) simulations during the passage of typhoons. The maximum increment in peak SWH reached 3.0 m and 5.0 m in the deep ocean for Super Typhoons Maria (2018) and Lekima (2019), respectively if the *Rtrs* was increased from 4 × *Rmax* (radius of the maximum wind) to 7 × *Rmax*. The SWHs induced by the typhoon winds in the surf zone were more sensitive to different wave-breaking formulations used in the wave-circulation model. The maximum difference in peak SWH reached 2.5 m and 1.2 m for Super Typhoons Maria (2018) and Lekima (2019), respectively, when the wave-breaking formulations of BJ78 (proposed by Battjes and Janssen in 1978) and CT93 (proposed by Church and Thornton in 1993) were introduced to the wave-circulation model. The SWH simulations in the surf zone were insensitive to the wave-breaking criterion (*γ*) during the passage of typhoons. In shallow nearshore waters, the utilization of a constant *γ* for the wave-circulation model always produces peak SWHs that are smaller than those using *γ* based on local steepness or peak steepness.

**Keywords:** depth-induced wave breaking; wave-breaking formulation; wave-breaking criterion; shallow nearshore waters


**María Ángeles Castro 1, Miguel Antonio García 1, José Antonio Martín 1 and Francisco Rodríguez 1,2,\***

Received: 5 September 2019; Accepted: 27 October 2019; Published: 3 November 2019

**Abstract:** In recent works, exact and nonstandard finite difference schemes for scalar first order linear delay differential equations have been proposed. The aim of the present work is to extend these previous results to systems of coupled delay differential equations *X* (*t*) = *AX*(*t*) + *BX*(*t* − *τ*), where *X* is a vector, and *A* and *B* are commuting real matrices, in general not simultaneously diagonalizable. Based on a constructive expression for the exact solution of the vector equation, an exact scheme is obtained, and different nonstandard numerical schemes of increasing order are proposed. Dynamic consistency properties of the new nonstandard schemes are illustrated with numerical examples, and proved for a class of methods.

**Keywords:** delay systems; nonstandard numerical methods; dynamic consistency


3.3.3. Total Peroxidase Activity

The total peroxidase activity (TPA) was assessed at 72 h according to the method of Dana Iordăchescu and Ioan Florea Dumitru published in 1988 [71]. The size of the leaf fragments was 0.5 cm × 0.5 cm, while the petiole fragments were 1 cm in length. As a general control, unwounded shoots were used. In this case, our experimental hypothesis was based on the knowledge that wound stress is already expressed during in vitro cultivation of plants, according to various authors [72,73]. Moreover, it has long been observed that a burst of phenolic compounds is generally released at the wound sites of plants, and that this process is immediately followed by the activation of membrane-bound peroxidases that are required to initiate healing processes [65,74].

• Effects of 2,4-D:

After 72 h, the petiole fragments treated with 2,4-D exhibited a 1.84-fold increase in TPA compared to control shoots (i.e., intact shoots). In this case, the auxin and the supplemented cytokinin clearly contributed to an increase in TPA to initiate cell proliferation, as the first and most obvious event was recorded four weeks later. In the case of control petioles, we found a 1.6-fold decrease in the TPA activity compared to explants in the presence of auxin, suggesting that auxin activated peroxidase activity in the petiole fragments. In fact, upon the increase in TPA activity at the wound site, cell proliferation began as an essential process for the survival of the plant inoculum. The calli or fragments of plants can further act to continue the plants' morphogenesis [10]. In the case of leaf fragments, 2,4-D induced a 2.45-fold increase in TPA activity compared to intact leaves and a 1.25-fold increase compared to the control leaf fragments. Due to the larger leaf surface for wounding, the release of phenolic compounds [27–29] might have been responsible for the activation of enzymes with peroxidase activity, which should greatly enhance the wound stress response of leaf fragments in the presence of 2,4-D.

• Effects of salicylic acid:

For petiole fragments, SA induced a 4.26-fold increase in TPA activity compared to petioles taken from intact shoots and a 1.46-fold increase compared to the control petiole fragments. This value was more than double the effect of 2,4-D at the wound site and is consistent with previous scientific results [60]. In the case of leaf explants, SA induced a ~1.2-fold increase in TPA activity compared to the intact leaves but, conversely, a 1.6-fold decrease compared to control leaf fragments. Here, SA induced TPA activity at half the rate of 2,4-D. Moreover, the TPA value in fragmented leaves was almost equal to that in intact leaves from intact shoots. It can be concluded that the exogenously applied SA may minimize the expression of TPA in syngonium at the leaf level, but it can induce overexpression in the fragments of petioles.

• Effects of 2,4-D and salicylic acid:

When both compounds (2,4-D and SA) were exogenously applied, it appeared that the TPA activity was similar to that obtained in the fragmented petioles or leaves subjected only to 2,4-D. Thus, the effects of SA appeared to be hidden by the presence of 2,4-D.

#### 3.3.4. Callus Initiation from Fragmented Petioles and Leaves

The entire experiment was observed continuously for 8 weeks. In the first three weeks, leaf and petiole fragments subjected to 2,4-D became etiolated compared to controls and those treated with SA, which remained green. All petiole fragments subjected to SA developed hypertrophy at the cut part of the petiole following the circular cambium, which also spread along the entire length of the fragment. However, some of these samples only developed callus at the wound site—white and friable, in small quantities—and later in the cambial tissue as well. Among the two types of explants, only petiole fragments were able to produce a proliferative callus for syngonium at the basipetal part of the petiole, in a similar manner to that described for other species [62]. In contrast, based on the conditions of this experiment, it was not possible to induce callus formation at the leaf fragment level. Moreover, other authors were successful in initiating and studying callus formation based on a specific hormone balance [46,49,51–53,56,63]. However, it should be noted that a burst of phenolic compounds may also affect callus formation [75].

In another experiment conducted by Jin Cui and collaborators on the same cultivar but under different culture conditions (i.e., different plant growth regulators and different balances), the proliferation of callus was also possible at the leaf level [56].

Re-evaluating these results, it can be concluded that wound stress is also responsible for a clear sequence of events, i.e., increased peroxidase activity, followed by the expression of hypertrophy at the wound sites and, subsequently, by callus development. Hormone balance and mineral composition are the major inductors of callus formation, and different organ fragments may respond differently. The lack of callus formation in fragmented leaves under our experimental conditions may have been due to the culture medium composition, which was not supportive of the expression of hypertrophy, although it was proven to express the highest peroxidase activity. This subject should be of interest when investigating callus initiation for different purposes [46,49,51–53,56,63].


*3.3.2. Sn0.97Cd0.03Se*

As an alloy with contrasting behavior, we show the transport properties of Sn0.97Cd0.03Se (**Figure 8b**, **d**, **f**). The cadmium-doped SnSe is the system with the highest Seebeck coefficients in this study (**Table 1**). Conversely, the electrical conductivity is very poor at RT, but there is an extraordinary increase with temperature, as illustrated in **Figure 8** on the right (middle panel). Both the large Seebeck coefficient and the low electrical conductivity indicate very low charge concentration. The steady decrease of the Seebeck coefficient (highest at 500 K with 570 μV/K) accompanied by the rapid increase of the electrical conductivity is another manifestation of the Pisarenko relationship [6]. The exponential temperature dependence of σ indicates a gap of Eg = 0.6 eV. This is very similar to the intrinsic gap of SnSe. We can conclude from this study that Cd is unlikely to be a useful dopant for SnSe since its power factor is rather small, probably because of a lack of effective charge transfer (i.e. doping).


**6. Conclusions**

Long life is a topic of great interest in medicine and among the general public. The increase in life expectancy has led the very old to become the fastest growing segment of the population in several countries.

#### *Update in Geriatrics*

Centenarians are subjects living 100 years or older. The majority of centenarians are females and a very small fraction of centenarians are supercentenarians.

Most centenarians have managed to avoid, postpone, or overcome the important age-related and life-threatening diseases (e.g., ischemic heart disease, stroke, chronic obstructive pulmonary disease, cancer, respiratory infection, type 2 diabetes, osteoporosis, and dementia).

The continuous increase of the number of centenarians has worldwide practical implications including profound impact on intergenerational interactions and significant financial challenges for any society, especially in relation to medical expenses.


**6. Roof-top solar photovoltaic with battery energy storage system**

Considering the same RTPV installed capacity of 200 W per residential home. In addition to this, it is assumed that each home is equipped with a battery which has 600 W of dispatchable power; an overview of this connection is seen in **Figure 5**. This inclusion of BESS is limited only by its charge and discharge rate. Based on the available power generated by a 200 W RTPV source, it was then assumed that the battery would reasonably charge and discharge at less than 7 amps.

The network normal (P) is the load profile for the study. **Figure 6** shows the comparison of RTPV with and without BESS. During daylight hours 80 W of power is allowed to charge the battery while the remaining power supplies the customer or overflows onto the electrical grid. During the evening peak period, the same 80 W is dispatched from the stored energy over a period of 7.5 hours.

As demonstrated in **Figure 6**, RTPV including BESS can be seen to benefit both the Utility and Customer. The Utility benefits by the reduced voltage regulation and thermal violations which are experienced during the evening peak, and the

**Figure 5.** *RTPV with battery storage [15].*

*Battery Energy Storage Systems and Rooftop Solar-Photovoltaics in Electric Power Distribution… DOI: http://dx.doi.org/10.5772/intechopen.99248*

**Figure 6.**

*Active power of NB36 with 200 W RTPV penetration and BESS, considered for a winter weekday.*

Customer benefits by an overall reduction in power consumed. From this analysis, it is seen that the Utility loses 5.76% in revenue. While the customer's power consumption, referring to data in **Figure 4**, is reduced by 5.8%. Further investigation of **Figure 4** shows a negligible difference of power consumed if the customer has RTPV alone or including BESS.

#### **7. Varied levels of BESS**

While the results from the above analysis speak of how the Utility and Customers are affected, there still raises the fundamental concern related to the constraints experienced on NB36. At normal, when the load is peaking, it's found that the minimum Voltage is 92.7% and the network is thermally loaded to 102%. The analysis shows that if customers provide back-feeding, from their stored energy, during peak times, it will reduce the constraints on the network. Analysis has considered the back-feeding of 80 W, following the discussion above, and 250 W for each residential customer.

**Figure 7** reflects the effect of the voltage along the backbone of NB36, also showing the ability of BESS to dispatch power at 80 W and 250 W. It can be seen that with an increasing ability of the BESS to dispatch power, it alleviates the voltage constraint by <1% at 80 W, and 2.1% at 250 W.

**Figure 8** shows that the normal network condition exceeds the rating of the conductor at peak loading.

With BESS dispatching power we see that the thermal constraint is reduced from 102–94% when 80 W of power is dispatched, and improved further when 250 W is dispatched to 79.8%.

#### **Figure 7.**

*NB36 minimum voltage at peak (winter weekday – 18:30).*

**Figure 8.** *NB36 thermal rating vs. load current.*

#### **8. Power dispatched, consumed and its effect on technical losses**

**Figure 9** shows the network sending power, consumed power (including with RTPV, and RTPV+BESS). The difference between the sending power and consumed power is attributed to the technical losses resulting from transmitting power down the network.

Technical losses are calculated as a percentage of the total load of the feeder including both no-load and load losses. The results from the analysis help to obtain a view of the network losses; this can be used as a basis for historic trending and benchmarking and can be used as one of the triggers for network strengthening. Therefore, this statistic also aids the conditioning of the priority ranking criteria. DER's may have a significant effect on network losses.

*Battery Energy Storage Systems and Rooftop Solar-Photovoltaics in Electric Power Distribution… DOI: http://dx.doi.org/10.5772/intechopen.99248*

#### **Figure 9.**

*Sending active power, power consumed by customers and P losses, considered for a winter weekday.*

A generator can lower or increase losses, depending on its location and the network configuration [16].

The technical losses vary slightly between the different options analyzed:

#### **9. Conclusion**

This chapter has demonstrated the benefit of roof-top solar photovoltaic and/or including battery energy storage systems. It offers relief for constrained networks in dense and radial distribution systems. While optimization techniques can be used to reduce violations, these are still limited do not provide effective short-term solutions when dealing with constrained networks in dense and radial distribution systems. Battery energy storage systems (BESS) and solar rooftop photovoltaics (RTPV) are a viable distributed energy resource to alleviate violations which are constraining medium voltage (MV) networks. The results show the following:

#### **9.1 BESS only**

This option does not benefit the customer, as the batteries require grid connection to charge and discharge, not to mention efficiency losses. If discharging the stored energy occurs during peak periods, this can benefit the utility by reducing the peak violations. Therefore, it is recommended that a tariff/time-of-use incentive is introduced to motivate customers for BESS only installations.

#### **9.2 RTPV only**

This is an excellent way for a customer to reduce his overall electrical utility bill. Unfortunately, the utility is negatively affected by the reduced sale of electricity.

#### **9.3 RTPV+BESS**

The results show that with the addition of RTPV including BESS the utility still loses revenue. However, with the addition of BESS, Utilities have the ability to reduce technical violations during peak periods. Installing BESS, in this manner, shows no benefit to the customer. Therefore, it is recommended that a tariff/timeof-use incentive is introduced to motivate customers for RTPV+BESS installations.

#### **9.4 Varied levels of BESS**

Though RTPV inclusive of BESS reduces the violations on the network; it can be seen from **Figures 7** and **8** that voltage and thermal violations persist for this specific network. Therefore, it is necessary to consider an increased installation of BESS. 250 W of dispatchable power offered the most appropriate quantity of power to alleviate the violations. The effect on technical power losses: seen in **Figure 9**, shows a constant 5.41%–5.49% of technical power losses for each of the above considerations. This demonstrates that technical losses are similarly proportioned to its source sending power when conducting analysis on the various installation types relating to RTPV/ BESS. While these amounts are typical and expected for a reticulation network – losses will defer due to topology, loading and design of networks.

For Utilities and Municipalities, the extent of challenges encountered when considering large scale installations of RTPV would be related to availability and visibility of data for adequate analysis. Current standards do not address the practical design solutions needed for all variations within customer installations and expectancy from RTPV. This can result in non-standard customer installations which will lead to undesirable impacts on the source and shared utility power systems.

Visibility and compliance of electricity supply regulations are required at the point of supply which is conditioned to be met at the point of supply and not the point of generator connection which is embedded in the customer's installation. Therefore, smart metering systems with predefined charge and discharge times/durations need to accompany RTPV installations. This requires an evaluation of currently employed metering technology, to ensure they accommodate these operational scenarios.

The cost for an electrical system, in which a customer is partially or completely off-grid, will still be attributed to the Utility to ensure the security of supply. Hence Utilities should take the lead in this segment of small-scale embedded generation to remain viable and relevant. Utilities may consider engaging suppliers to carry out maintenance and/or repairs during initial warranty periods.

Data visibility, both topological and metered data, is crucial for power system analysis, and need to be available and validated from time to time to cater for load growth. Utilities must ensure competent staffing for data acquisition and analysis. *Battery Energy Storage Systems and Rooftop Solar-Photovoltaics in Electric Power Distribution… DOI: http://dx.doi.org/10.5772/intechopen.99248*

There may be a lack of data validity for non-telemetered devices, such as unknown tap positioning of reticulation transformers, conductors or cables which has been replaced with different ratings and types. This can be challenging when configuring simulation models.

For this study analysis was given to the network classification that was predominately found with residential type loads, which has been identified as C2 TZ2 type. This study, therefore, adopted a common sending voltage regulation set point of 1.03pu applied at reticulation busses. The voltage limit allowable for a normal condition is 95.5% and for abnormal conditions are 93.5. From an operations view, the abnormal limit is the benchmark to be adhered to, while network planners work with the normal limit of 95.5%.

Continuous network changes due to rising electrification and illegal connections discredits network analysis and requires more frequent update of simulation models. Pre-existing RTPV installations may pose a problem if they do not meet compliance requirements. Utilities need to conduct surveys of pre-existing RTPV installations and update its electrical connection to the power grid.

It is recommended that during site visits, meter re-programming including 'time-of-use' tariffs and amendments to supply agreements, can be implemented. Presently, there is no information on households which has become "self-suppliers" due to lack of a registry for small-scale embedded PV plants. This will assist in ensuring that customer contributions to unbalance be limited to 1% voltage unbalance at the point-of-common coupling.


*2.4. Electrochemical Measurements*

Cyclic voltammetry (CV), galvanostatic charge-discharge (GCD), and electrochemical impedance spectroscopy (EIS) of single electrodes were performed using a potentiostat (VSP, Bio-Logic). The electrolyte is a 1 M H2SO4 aqueous solution that purged N2 gas for 20 min. Measurements are conducted through a three-electrode system. A reference electrode was a saturated calomel electrode (SCE), and a counter electrode was a platinum wire. For voltammetry measurements, the cell potential was scanned at 5 mV/s and 20 mV/s. When the scan rate was 5 mV/s, it was measured for 5 cycles, and when it was 20 mV/s, it was measured for 4990 cycles. Galvanostatic charge-discharge behavior was measured in current density, from 0.5 to 20 A/g. EIS was measured from 100 kHz to 100 mHz.


**3.2 Anti- epileptic effect**

Epilepsy is one of the most common, serious neurological conditions, affecting more than 50 million people worldwide. The hydroalcoholic extract of *P. vera* demonstrated a significant anti-epileptic effect in pentylenetetrazole (PTZ) chronic induced kidding in male rats [57]. The epileptic condition was induced by PTZ (40 mg/kg, IP), and the induced condition was significantly inhibited by the extract of *P. vera* at the tested doses of 50 and 100 mg/kg. The inhibition of chronic induced seizure indicate that *P. vera* is a significant antiepileptic. The petroleum ether extract of *P. integerrima* attenuated the PTZ-induced jerks in zebrafish and mice models at the dose of 50 and 100 mg/kg. The antiepileptic effect was further confirmed through maximum electroshock (MES) in a rat model. The tested extract significantly attenuated various aspects of induced jurks [58].