**12.2 Fissurectomy**

Fissurectomy as a treatment for anal fissures in children was found successful only when combined with postoperative laxative therapy (Lambe et al., 2000a). An important part of their technique was the use of stay sutures to avoid the need for an anal retractor, thereby preventing stretching of the internal anal sphincter. A triangular part of the anoderm is excised along with the fissure itself. A good and reliable operation, but leaves behind a large and uncomfortable external wound, which takes a long time to heal.  Lateral internal sphincterotomy (LIS) has been found to be the preferred operation. LIS can be performed using either the open or closed technique , the method of Notaras, dividing the IAS via a small stab wound (Notaras, 1971). Equal success has been reported with open or closed lateral sphincterotomy for acute and chronic anal fissures in children. A systematic review on the treatment of anal fissure was published by Steele & Madoff in 2006. Current concepts in anal fissures were reported by Ayantude et al. (2006) in the same year, involving a literature search from 1970 to 2004.

Chronic anal fissures tend to be refractory and are usually reluctant to heal with conservative treatment.  A very large series of adult patients was published by Lysy et al. in 2006. Prolonged periods of treatment were necessary and 384 patients were healed (84.4%) by the end of four months. Older age and longer time interval between symptom appearance and treatment negatively affected fissure healing. The explanation for the latter was that longer time exposure of the fissure area to

Nifedipine Gel with Lidocaine in the Treatment

Fig. 4. Clinical presentation (percentage)

bases and overlapping skin edges.

**14. Special situations**

results (12 improved).

**15. Conclusions**

require hospitalization.

of Anal Fissure in Children: A Pilot Study and Review of the Literature 65

1.  Frequently, they are multiple or off the midline, and often coexist with other pathology (Sangwan et al., 1996). They can be locally aggressive, progressing to form deep ulcers with granulating

2. HIV/AIDS: Anal fissures maintain their typical appearance, but have poor wound healing (Lord, 1997). Barrett et al. (1998) reported their experience with perianal disease in 260 HIV-positive patients, 32% of them with anal fissures (Barrett et al., 1998). Eighteen patients underwent sphincterotomies. Viamonte et al. (1993) reviewed the treatment of 33 HIV-positive fissure patients. Thirteen underwent LIS, with excellent

Topical 0.2% nifedipine with lidocaine appears today as the most efficient mode of treatment for anal fissures in children, with a significant healing rate and no side effects. It is safe and effective, prevents the evolution of acute anal fissures in children to chronicity, avoids surgical procedures in the great majority of cases, avoids complications and does not

inflammation and ischemia and subsequent fibrosis may compromise the healing process. After recurrence, patient education for self and prompt retreatment was found to improve outcome. Chronic anal fissures are caused by internal sphincter hypertonia, which leads to reduced blood flow and tissue hypoxia, with consequent healing failure. A cautious surgical approach is required to treat those who do not respond to medical treatment, and should include excision of the fissure along with its sentinel tag and internal sphincterotomy at the base of the ulcer (Cohen & Dehn, 1995; Lambe et al., 2000b). The wound is left open and should heal in 7-14 days without scarring. Local reconstruction with advancement flaps is a relatively new and effective adjunct to chronic fissure excision. Practice parameters for the management of anal fissures from the American Society of Colon and Rectal Surgeons were reported by Orsay et al. (2004) and a very extensive review on the diagnosis and care of patients with anal fissure was reported by the American Gastroenterological Association (Madoff & Fleshman, 2003).


5. Solid lipid nanoparticles for liver targeting

Though targeted delivery of a drug to the liver is a challenging task, still, it is an interesting approach in the treatment of various liver disorders. In the treatment of liver disorders, drugs targeting to the liver, face irresistible obstacles from various physiological barriers and processes like uptake by the reticuloendothelial system, mechanical entrapment by the pulmonary vascular bed, and opsonization process [43].

Numerous approaches are being proposed to enhance bioaccumulation/ biodistribution of drugs to liver and hepatocytes. These approaches include both active targeting as well as passive accumulations of nanoparticulate formulation due to ligand (carbohydrate, peptide, antibodies conjugation) conjugated nanoparticles. Recently, liver targeted deliveries of drugs by the SLNs are gaining much attention in the treatment of various types of liver disorders. Thus, various liver targeting strategies using SLNs are enlightened below.


**Author details**

Cory M. Willis and Stephen J. Crocker\*

\*Address all correspondence to<EMAIL_ADDRESS>
Department of Neuroscience, University of Connecticut School of Medicine, Farmington, CT, USA


**4. Practical tips to minimize the arrhythmic complications**

which leads to more advancement of PICC line. This can be avoided by using rope method of measurements (can use a nasogastric tube or any flexible tubes) along the course of veins applicable to lower and upper extremity veins.

be due to the migration of catheter until proved otherwise. So check for migration, verify the external length and immediately withdraw the catheter. Confirm the position with an X-ray, check for kinking, curving and looping which is potentially dangerous as it can

Central Venous Catheter-Induced Cardiac Arrhythmias in Neonates

http://dx.doi.org/10.5772/intechopen.74559

173

**12.** Vigilantly look for catheter migration whenever the critical neonates are on high-frequency

**13.** Take precautions while handling for ultrasonography, daily weight check, the neonatal transfer for any cause, pre and post-operative periods and any invasive procedures which

**14.** Ultrasound-guided insertion of central lines (PICC or UVC) is more reliable, safe and

**15.** While inserting the PICC line turn the head of the neonate toward the ipsilateral side of

Central line insertion is a very common bedside procedure in NICU setup, it is not uncommon to encounter complications. Although there are various complications related to PICC line insertion, arrhythmic complications are preventable by exact measurements along the venous course, avoidance of willful over advancement of the catheter, prevention of migration and early detection of migration and adjusting the catheter tip properly. Ultrasoundguided insertion of catheter and placement of catheter tip using real-time ultrasonography is a welcoming advancement to minimize arrhythmic complications. Worldwide, the reported

insertion as it prevents the entry of PICC line into the jugular veins.

incidence of arrhythmic complications published in the literatures are few.

induce pericardial tamponade.

need more manipulation of neonates.

effective when compared with the blind procedure.

ventilation.

**5. Conclusion**

**Abbreviations**

CVC central venous catheter

UVC umbilical venous catheter

SVT supraventricular tachycardia NICU neonatal intensive care unit

AF atrial flutter

IVC inferior vena cava SVC superior vena cava

PICC peripherally inserted central catheter

The interaction between the neonatal posture, and radiological catheter position is well explained in advances in neonatal care.

**Table 5.** Ideal positions for X-rays pertaining to different sites of PICC line [10].

be due to the migration of catheter until proved otherwise. So check for migration, verify the external length and immediately withdraw the catheter. Confirm the position with an X-ray, check for kinking, curving and looping which is potentially dangerous as it can induce pericardial tamponade.


**8. Conclusion**

First hour following birth for mature newborn is without doubt the most critical hour in life of human beings. During this time, a lot of changes happen. When woman gives birth, all the hard work she does generate changes in the chemistry in the brains. It makes women want to nurture her child. These hormones also cause the uterus to contract, shrink and stop bleeding.

Based on decades of evidence, the World Health Organization and United Nations Children's Fund [6] recommended that all healthy mothers and babies, regardless of feeding preference and method of birth, have uninterrupted skin-to-skin care beginning immediately after birth, lasting for at least an hour.

There is still a lot of unnecessary interventions in the first hour after birth in many maternity hospitals. Routine procedures are being carried out starting from early cord clamping to vitamin K injection, eye prophylaxis antibiotic ointment, navel prophylaxis, foot and hand printing, weighing, measurements and bathing and others. All health-care providers should know that immediate skin-to-skin contact is the best way for a newborn and mother to bond. Healthy newborns should be placed in 'skin-to-skin' contact with the mother until the first round of breastfeeding is established. Skin-to-skin care means placing dried, unclothed newborns on their mother's bare chest, with warmed light blankets or towels covering the newborn's back. Women who have a planned or unplanned C-section would not be in the ideal position for intimate bonding right away. Baby could be taken to a warming table for a quick assessment first. Authors [18] claim that there is no reason why stable mothers should not have the experience of skin-to-skin contact after cesarean births, to collect the same short- and long-term benefits of it. Even from psychological point of view, it helps them mourn the loss of a normal vaginal birth.

The first hour should be focused on baby's first breastfeed and mother-baby and family bonding. The manner in which a new baby is welcomed into the world during the first hours after birth may have short- and long-term consequences.


**Identification and Functional Characterization of Novel Venom Components**

Special Issue Editor **Steven D. Aird**

MDPI • Basel • Beijing • Wuhan • Barcelona • Belgrade • Manchester • Tokyo • Cluj • Tianjin

*Special Issue Editor* Steven D. Aird Technical Editor Japan

*Editorial Office* MDPI St. Alban-Anlage 66 4052 Basel, Switzerland

This is a reprint of articles from the Special Issue published online in the open access journal *Toxins* (ISSN 2072-6651) (available at: https://www.mdpi.com/journal/toxins/special issues/novel venom components).

For citation purposes, cite each article independently as indicated on the article page online and as indicated below:

LastName, A.A.; LastName, B.B.; LastName, C.C. Article Title. *Journal Name* **Year**, *Article Number*, Page Range.

**ISBN 978-3-03936-467-1 (Pbk) ISBN 978-3-03936-468-8 (PDF)**

Cover image courtesy of Steven D. Aird.

c 2020 by the authors. Articles in this book are Open Access and distributed under the Creative Commons Attribution (CC BY) license, which allows users to download, copy and build upon published articles, as long as the author and publisher are properly credited, which ensures maximum dissemination and a wider impact of our publications.

The book as a whole is distributed by MDPI under the terms and conditions of the Creative Commons license CC BY-NC-ND.


**Preface to "Measurement Uncertainty"**

This book treats about measurement uncertainty, both from the theoretical point of view and from the practical point of view. From the theoretical point of view, contributions are present which propose more general mathematical methods to handle measurement uncertainty. On the other hand, from the practical point of view, contributions are present which show how measurement uncertainty is considered in the specific applications. The aim of this book is to provide a wide overview on this very important topic.

> **Simona Salicone** *Editor*


*4.1. Conformational Clustering*

The conformational ensemble of the studied construct encompassing N-WASP domain V and previously generated by MD simulations with the Amber-03ws force field [29] was clustered with the GROMACS tool *gmx cluster* using the *gromos* method [35] and a RMSD cutoff of 0.5 nm (computed over the mainchain atoms). We obtained 2467 clusters and decided, for subsequent protein–protein docking calculations, to keep only the 527 most populated ones, which represent 50% of the 40,000 conformations sampled by MD simulations. To verify that the 527 clusters are representative of the overall conformational ensemble, we compared the residue probabilities to be in *α*-helix and the distributions of gyration averaged over the 40,000 conformations or the 527 cluster structures. As shown in Figure S2, the probabilities to form *α*-helices of the 527 clusters and of the whole conformational ensemble are almost identical, and the protein radius of gyration has similar distributions when computed over the sub-ensemble of representative structures or over the 40,000 conformations. This indicates that the selected 527 conformers are locally and globally representative of the whole conformational ensemble of N-WASP domain V.


**2.3 Case study**

74 Recent Advances in Nanofabrication Techniques and Applications

account, the number is 1, otherwise 0. Likewise, the second, third and fourth bits denote the

Conf. 2 (1,0,1,0)

799.5 × 792 N/A N/A N/A N/A 810 × 808.5 7.09 6.97 N/A N/A 819 × 808.5 7.37 7.43 N/A N/A 829.5 × 825 7.52 - N/A N/A 840 × 825 7.16 - N/A N/A 849 × 841.5 7.24 - N/A N/A 859.5 × 858 7.11 - N/A N/A 870 × 858 - - 7.15 N/A 879 × 874.5 - - 7.23 7.15

A number in each column denotes the delay time with the least area "N/A" means that the given areas were infeasible to place and route the circuit with the place-and-route tool. For example, the least area and the delay time for the area were obtained as 810 × 808.5 and 7.09 ns respectively in Conf. 1. The least areas in Confs. 1, 2, 3, and 4 were 810 × 808.5, 810 × 808.5, 870 × 858, and 879 × 874.5 respectively. About 14% area increased when the mirror-Y and the mirror-XY were forbidden. Theoretically speaking, delay time decreases under a case in which one can use a larger place-and-route area. Delay time in a column of the table is expected to decrease downward but it did not. This is because the CAD tool is based on approximate algorithm. Comparing the two values of Conf. 2, 6.6% delay time

Conf. 1 is the configuration in which all the four cell directions are available and is supposed to be best with regard to area and delay time among the configurations because its design space includes design space of the other configurations. In other words, any layout based on Confs. 2, 3 or 4 can be realized by Conf. 1. The results which the CAD tool reported does not straightforwardly reflect this supposition because the layouts obtained by the CAD tool are approximate solutions, e.g. the delay time of Conf. 2 (6.97 ns) was shorter than that of Conf.

Conf. 2 is the configuration in which the horizontal flippings are removed from Conf. 1. In other words, the mirror-X and mirror-XY directions are not taken into account in Conf. 2. There was no great difference of delay times among Confs. 1 and 2. Horizontal flipping

Conf. 3 is the configuration in which the vertical flippings (mirror-Y and mirror-XY) are removed from Conf. 1. Experimental results show that the vertical flipping of cells had little influence on delay time of the chip and it had some influence on the area. Comparing Conf. 3 with Conf. 1, about 14% area increased. This is because the gaps between cell areas were added by eliminating vertical flipping of cells and each cell area got to own its own power

Conf. 4 is the configuration in which any flipping cells are forbidden and only a basic direction of cells is available. Comparing Conf. 4 with Conf. 1, the differences of the delay

Delay times for four cell directional variations [ns], (basic, mirror-X, mirror-Y, mirror-XY)

> Conf. 3 (1,1,0,0)

Conf. 4 (1,0,0,0)

existences of the mirror-X, mirror-Y and mirror-XY directions respectively.

Conf. 1 (1,1,1,1)

Table 1. Delay times for four cell directional variations

increased while place-and-route area increased.

1 (7.09 ns) as the place-and-route area was 810 × 808.5!

seems not to be so effective to reduce delay time and area.

and ground lines.

Chip area

In this section, a case study is shown for five cases to examine the relation between the number of EB shots and how to select cell objects to place on characters. The five cases are described in Table 2. We developed the cell selection software described in Section 2.1.1 with a commercial mathematical optimization engine, ILOG CPLEX 9.0 (ILOG, 2003). Every optimization process finished within a second.

Table 2. Cell directional variations for experiments

The specification of the CP equipment for which we assumed is shown in Table 3. Two benchmark circuits were used to examine their numbers of EB shots under the five cases. The description for the benchmark circuits is shown in Table 4. Note that the cell library is from academia and comprises fewer kinds of cells than that from industry.

Table 3. Specification of CP/VSB equipment

Character Projection Lithography for Application-Specific Integrated Circuits 77

approximately under Cases 2 and 4 by assigning the equal number of characters to each

Comparing the number of EB shots of Circuit 1 under Case 2 with that under Case 4, 44% reduction of EB shots was achieved. Likewise, comparing the number of EB shots of Circuit 2 under Case 2 with that under Case 4, about 26% reduction of EB shots was achieved. It was experimentally found that the elimination of cell directions is quite effective to reduce EB shots. It was experimentally found that the elimination of horizontal flipping reduced the much number of EB shots effectively while it has small impact on area and delay time of

In this section, we proposed an ILP-based cell library development methodology to reduce the number of EB shots. All optimization processes finished within a second. More than 3.85% reduction of EB shots was achieved only by distinguishing between the differently

We examined the influence of cell directions on both area and delay time of the circuit. It was experimentally confirmed that both of the horizontal and vertical flipping of cells had little influence on delay time of chips. The horizontal flipping had little influence on area while the vertical flipping had some influence on area. This examination helps which cell

The forbiddance of horizontal flipping caused little deterioration of area while 25.6% reduction of EB shots. It was found that the forbiddance of horizontal flipping was effective to reduce the number of EB shots while it deteriorated little area and delay time of chips. The forbiddance of vertical flipping caused 13.9% increase of area while it caused less than 1% increase of delay time. The forbiddance of vertical flipping should be determined with taking a tradeoff between area and EB shots into account. For many chips of the state of the art, cells are placed so "loosely" that the deterioration of area caused by forbiddance of multi directions of cells might have less impact on area. The relation between cell directions

Character size optimization techniques were studied for character projection (Sugihara, 2006c, 2007b, 2010). We first presented an idea to optimize a character size for higher projection throughput (Sugihara et al., 2006c). We presented a character size optimization by enumerating all possible character sizes and generating a cell library for every given character size (Sugihara et al., 2006c). Next we presented a cell library development methodology in which a character size and a set of cells were optimized at the same time (Sugihara et al., 2007b). We also presented a character size optimization technique for multicolumn-cell projection equipment. In this section, we focus on the first work (Sugihara et al.,

In Section 2, the size of characters on CP masks was given and treated as a constant because of the restriction which attributes to character projection equipment. Cells used in a design were, consequently, partitioned to fit the constant size of characters by intuition. In this

and EB shots in design of such chips should be further examined as future work.

**3. Character size optimization for higher projection throughput**

directions of cells.

**2.4 Conclusion**

mirrored cells whose functions are identical.

2006c) for a simple explanation.

**3.1 Character size optimization problem**

direction should be implemented on CP aperture masks.

chips.

Table 4. Benchmark circuits

The EB shots under the five cases were sought out by solving mathematical problem instances and are shown in Table 5. In the table, the parenthesized values show the numbers of cell objects. The areas and delay times of Circuit 2 are shown in Table 6. In our experiment, areas and delay times of Circuit 1 were not examined because the benchmark circuit was not logic-synthesizable.

Table 5. EB shots and cell objects

Table 6. Area and delay time of Circuit 2

According to Table 5, as the number of cell objects increases, in other words, the number of cell directions increases, the number of EB shots increases. This is because the reduction of cell directions enables more cell functions to be on a CP aperture mask and to be projected with the CP. The area of Circuit 2 under Case 1 was largest among the five cases as shown in Table 6 because only a single direction, that is a basic direction, was adopted for place-androute. This was because the gaps between cell areas came to arise and each cell area got to own its own power and ground lines. Theoretically speaking, the design with the four cell directions should be best among the five cases with regard to area and delay time. Similarly, the design with the two cell directions should be intermediate. The experimentally obtained values of areas do not reflect this supposition. The delay time of Circuit 2 under Cases 2 and 3 was found best. This is because the CAD tool returned approximate solutions of layout and happened to result against the supposition. Note that the values shown in Table 1 and Table 6 are nothing more than the ones the CAD tool reported. If a design obtained with the two cell directions is better than a design obtained with the four cell directions, the design of two-cell direction may be adopted as a design of four-cell direction.

Comparing the number of EB shots of Circuit 1 under Case 4 with that under Case 5, 3.85% reduction of the number of EB shots was achieved. The difference in the numbers of EB shots was caused by solving the problem instances exactly or approximately. The optimal sets of cells was selected exactly under Cases 3 and 5 while sets of cells was selected approximately under Cases 2 and 4 by assigning the equal number of characters to each directions of cells.

Comparing the number of EB shots of Circuit 1 under Case 2 with that under Case 4, 44% reduction of EB shots was achieved. Likewise, comparing the number of EB shots of Circuit 2 under Case 2 with that under Case 4, about 26% reduction of EB shots was achieved. It was experimentally found that the elimination of cell directions is quite effective to reduce EB shots. It was experimentally found that the elimination of horizontal flipping reduced the much number of EB shots effectively while it has small impact on area and delay time of chips.


**8. Natural antioxidants: role against aging**

Role of antioxidants in quenching ROS has been appreciated for a long time. Recently, a 15 yearlong study showed that ROS formation could be lessened by 10% in people aged 45 or above by simply having an antioxidant rich diet [71].

Antioxidants are compounds that quench the free radicals by providing an electron and generating stable compounds that are less harmful to a cell. In this manner they inhibit the train of events that occur when free radicals oxidize other biomolecules. Antioxidants can be classified into different categories based on enzymatic/ non-enzymatic activity, occurrence, solubility although other miscellaneous molecules displaying antioxidant properties have also been reported. Examples of certain regular antioxidants found in a cell involve Superoxide dismutase (SOD), Glutathione peroxidase (GPx) and Catalase (CAT). Non-enzymatic antioxidants include Vitamin E, C, thioredoxin, glutathione and melatonin. Further antioxidants such as minerals (Cu, Zn, Mn, Se), other vitamins, polyphenols and carotenoids are also derived from regular diet [72].

Usually, cells maintain a fine balance between ROS formation and their quenching by resident antioxidants. But, UVR shift the balance towards ROS formation resulting in a phenomenon called 'Oxidative stress', where resident antioxidants become incapable of quenching the high amount of ROS formed [73]. This state can however be circumvented by additional antioxidant supplementation. Drawbacks of synthetic antioxidants such as toxicity, low solubility, cost and a presumed negative outlook towards chemical products have increased the demand for natural antioxidants [74]. These antioxidants are naturally available in numerous sources like vegetables, fruits, spices, herbs, edible mushrooms among others [75]. Furthermore, beverages derived from these sources are also rich in antioxidants and have gained increased populace especially among younger generation.

Till date, several pure phytochemicals and plant-based compounds have been reported for their antioxidant and anti-photoaging activity. We have tried to summarize some important phytochemicals as potential antioxidant and anti-photoaging agents in **Tables 1** and **2**.

### **8.1 Beverages as source of antioxidants**

Various beverages, specifically mixed juices have abundant antioxidants as they are made using various fruits and vegetables [86]. Other beverages like coffee and

*Skin Aging: Implications of UV Radiation, Reactive Oxygen Species and Natural Antioxidants DOI: http://dx.doi.org/10.5772/intechopen.100102*

#### **Table 1.**

*Pure phytochemicals and their anti-photoaging properties.*

tea also include antioxidants and can help alleviate photoaging signs. Green tea is enriched in bioactive compounds like methyl-xanthins, flavon-3-ols and catechins such as (−)-epicatechin (EC), (−)-epicatechin-3-gallate (ECG), (−)-epigallocatechin (EGC) and (−)-epigallocatechin-3-gallate (EGCG). EGCG can inhibit erythema and neutrophil infiltration in the skin induced by UVR [87]. Drinking Green tea can ameliorate other photoaging markers like increased numbers of dermal cysts, sebaceous glands enlargement, vacuole formation and increased epidermal thickness. Green tea polyphenols are also elemental in decreasing MMP-2, 3, 7 and 9 levels along with reduced oxidation and carbonylation of proteins upregulated under UVR [88, 89]. Polyphenols and Chlorogenic acid found in coffee can help avoid free radical associated diseases. Reduction in pigmentation spots induced under UV was seen in Japanese women who consumed coffee at a higher rate [90]. *Coffee Arabica* leaf extracts are rich in caffeic acid, chlorogenic acid and phenols. These bioactive compounds help lower MMP-1, 3 and 9 expressions while elevating type I procollagen production. Further, a decrease p-ERK, p-JNK and p-p38 levels has also been shown thereby effectively blocking UV induced activation of key signaling pathways [91].

**Table 2.**

*Plant-derived phytocompounds having antioxidant and anti-photoaging activity.*

A diterpenoid, Atractyligenin found in Coffee silverskin has been shown to decrease ROS levels by 62% post UV treatment, modulate MMP-1, 2, 3 expressions, inhibit activation of MAPK pathways and increase Endo180 receptor expression [92].


*2.2. Intervention Design*

Based on the researchers' criteria, the company's regional director for health and safety recommended five skilled nursing facilities that did not yet have active HP programs and whose administrators were expected to be receptive to the participatory intervention process. We selected three of these facilities using a priori criteria to judge which were most organizationally ready for PIP [32]. Another three facilities with pre-existing, corporate-initiated HP programs were recommended by the regional director as control (NPHP) sites on the basis of their current activities and administrator commitment to the program [32].

In the PIP centers, team members were recruited from the entire workforce from volunteers responding to posters and management promotion of the program. Each team started with employees from various departments (clinical, dietary, housekeeping, laundry, maintenance, office/business) who met bi-weekly for one hour with two researchers. Initial team meetings involved identification of key issues in workplace health, psychosocial stress, and work organization.

The intervention began with intensive orientation of the PIP team (2–4 meetings per site over 1–2 months) to worker health and well-being, and Total Worker Health as a comprehensive approach. PIP teams identified issues of importance to members and discussed possible solutions or projects to address these concerns. Team members sought the opinions of their co-workers for program goals and specific activities. PIP team members communicated with individuals at various levels of their facility with updates and available activities (Figure 2).

Initially the researchers facilitated meetings, guided discussions, and assisted in framing presentations to the site administrator regarding a team's proposed project. Active facilitation involved the wellness champion and a research assistant attending all bi-weekly team meetings over 2 to 3 years, followed by monthly team meetings over 1 to 2 years, then quarterly telephone check about the program process with the wellness champion per site. The researchers provided technical assistance on a variety of topics, such as seminars on ergonomics in skilled nursing facilities and a food preferences survey to assist in developing programs for healthier food provision. Meeting minutes and activity logs were maintained by the researchers and utilized for ongoing process evaluation.

The goal was that participatory teams would move over time from co-governance to become independent, with support of the facility wellness champion. Thus, the study plan called for the researchers gradually to reduce our facilitation efforts over time. This was communicated to all participants at the beginning of the project. The phase-out period entailed 1–2 years of quarterly telephone check-ins with the wellness champion.

**Figure 1.** Timeline of the participatory intervention process and impact evaluation. HP, health promotion; PIP, participatory intervention program; CPH-NEW, CenterforthePromotionofHealthintheNewEnglandWorkplace.

**Figure 2.** Participatory Intervention Design within Centers.

#### *2.3. Data Collection and Analysis*

A mixed-method (convergent parallel strategy) approach utilized qualitative and quantitative data to examine the process and impact of the participatory OSH/HP or NPHP program in each facility. Results from the qualitative and quantitative analyses were triangulated with the researchers' direct experiences and knowledge of the organization to understand the process, impact, and sustainability of the participatory program.

Quantitative data included a brief baseline (pre-intervention) survey of the wellness champions about HP program activities in all six centers. We also conducted employee surveys in the six centers at baseline (2008–2009) and around the fourth year (2012–2013) of the participatory intervention. A self-administered questionnaire collected information on worker chronic disease history, health beliefs and behaviors, and perception of the work environment: physical and psychological job demands, decision latitude, and social support from supervisors and coworkers [8,33,34]. Work environment items included physical exertion, safety climate, psychological demands, decision latitude, and supervisor and coworker support. Psychological demands (two items), decision latitude (two items), physical exertion (five items), and supervisor (two items) and coworker support (two items) were selected from the Job Content Questionnaire (JCQ) [35]. The JCQ subscales have demonstrated good validity and acceptable internal consistency in large study populations from six countries [35]. Safety climate was measured with two items from Griffin and Neal [36] and two items developed by the investigators.

Most analyses compared employees in the pooled intervention (PIP) group to the pooled control (NPHP) group (three centers per group). Baseline (*n* = 645) and post-intervention (*n* = 649) prevalences were compared by cross-tabulation with chi-square statistics and mean values by t-test for independent samples. Cumulative incidence of self-reported chronic diseases was computed from baseline to post-intervention within each group and compared between PIP and NPHP with Fisher exact tests due to small numbers. Within-person changes from baseline to follow-up (limited to workers responding to both surveys) were examined using stratified cross-tabulation and paired sample t-test. All analyses were done with SPSS 22.0 (IBM SPSS, Chicago IL, USA).

Qualitative data included the meeting minutes and activity logs collected throughout the active facilitation period. Follow-up data were collected 3–4 years after the intervention began (2011–2012). Data types included: (1) focus groups with team members; (2) focus groups with other nursing home employees; (3) in-depth interviews with individual team members and wellness champions; (4) in-depth interviews with management (administrators and directors of nursing); and (5) in-depth interviews with supervisors (department heads and unit/office managers) [24].

Other evaluation materials included researchers' field notes on observed indoor spaces (employee lounge, break room, vending machines, and bulletin boards), outdoor spaces (employee picnic areas and gardens), and printed materials (employee newsletters, flyers, and informational literature) devoted to HP, OSH, or related activities or information. Researcher experiences and observations were logged after each field visit and consulted for purposes of this evaluation. Content analysis was performed using NVivo 9.0 software on transcripts from interviews and focus groups. We compared the themes that emerged across and within the six sites, focusing on (1) integration of HP and OSH, (2) comparative effects of the PIP and NPHP programs, and (3) sustainability of PIP as a model.

Metrics for comparison and evaluation included:


**8. Acknowledgement**

This work was supported by ITESI, CONACYT under thematic network RedNyN agreement I0110/229/09, and PROMEP agreement 92434. The first author thank to his wife and son for their time and patience to realize this study.

#### **9. References**

58 VLSI Design

three-terminals, they are modeled by a bipolar junction transistor (BJT) or a metal-oxidesemiconductor field effect transistor (MOSFET), the latter being the most common for VLSI

Fig. 11. Cross section of resistive gas sensors and biosensors: (a) sensors based on carbon

minute quantities of gas molecules (Jesse et al., 2006; Mahar et al., 2007).

The use of complex morphologies and structures based on composites containing carbon nanotubes and polymers in the design of gas sensors, has allowed the detection of polar and nonpolar gases making use of the change of dielectric constant to enhance sensitivity to

Graphene is exploited due to its inexhaustible structural defects and functional groups. These are advantageous in electroanalysis and electrocatalysis for electrochemical applications such as gas sensors and biosensors. Physisorbed ambient impurities by graphene such as water and oxygen can produce an effect similar to hole-doping and therefore a behavior similar to a *p*-type material (Traversi et al., 2009). Then, the graphene can be exploited as a sensing material for the design of chemical and/or biochemical sensors. When graphene is doped, well-identified localized states are added and band gap is introduced to the electrical properties generating an interesting alternative to design sensors

The main changes to be realized in the optimization of performance of gas sensors are the search of methods which allow us to synthesize identical and reproducible CNTs will give place to gas sensors with high quality and high performance, independently of the type of

In accordance with the review proposed here, CNTs are very attractive as base material to the design of components for VLSI Design. Chemical modifications of CNTs allow to the designer improve the selectivity of the electrical properties for the different applications. In the future, the use of hybrid materials where carbon nanotubes are involved will be a priority, given that the use of composite materials to design electronic devices, circuits and sensors requires multiple physical and chemical properties that a unique material cannot provide by itself. In the search for reducing electrical resistance presented by carbon nanotubes, different strategies have been developed to improve the efficiency of interconnection between devices based on carbon nanotubes and metallic electrodes used to lead the electrical bias to them. The implementation of digital and analog circuits with CNFETs or graphene nanoribbons will produce a great advance toward VLSI design using nanoelectronics. Still, hurdles remain as it was described in each section of the chapter.

systems (Zhao et al., 2008).

(Barrios-Vargas et al., 2011).

**7. Conclusions**

nanotubes, and (b) sensors based on graphene.

chemical functionalization required for the detection.

Carbon Nanotube- and Graphene Based Devices, Circuits and Sensors for VLSI Design 61

Ferry, D.K.; Goodnick, S.M. & Bird, J. (2009), *Transport in Nanostructures*, Second Edition,

Geim, A.K. & Novoselov, K.S. (2007), The Rise of Graphene, *Nature Materials*, Vol. 6, No. 3,

Geim, A.K. (2009), Graphene: Status and Prospects, *Science*, Vol. 324, No. 5934, pp. 1530-

Ginley, D. S. (2010), *Handbook of Transparent Conductors*, Springer, ISBN 978-1-4419-1637-2,

Giustiniani, A.; Tucci, V. & Zamboni, W. (2011), Carbon Nanotubes Bundled Interconnects:

Goel, A. K. (2007), *High-Speed VLSI Interconnections*, Second Edition, John Wiley & Sons,

Gruner, G. (2006), Carbon Nanotube Transistors for Biosensing Applications, *Analytical and*

Guildi, D.M. & Martín, N. (2010), *Carbon Nanotubes and Related Structures: Synthesis,*

Haruehanroengra, S. & Wang, W. (2007), Analyzing Conductance of Mixed Carbon

Hasan, S.; Salahuddin, S.; Vaidyanathan, M. & Alam, M.A. (2006), High-Frequency

Hatakeyama, R.; Li, Y.F.; Kato, T.Y. & Kaneko, T. (2010), Infrared Photovoltaic Solar Cells

Hayden, O. & Nielsch, K. (2011), *Molecular- and Nano-Tubes*, Springer, ISBN 978-1-4419-9442-

Hecht, D.S.; Hu, L. & Grüner, G. (2007), Electronic Properties of Carbon Nanotube/Fabric

Hierold, C. (2008), *Advanced Micro & Nanosystems Vol. 8 Carbon Nanotube Devices: Properties,*

Hong, S.W.; Banks, T. & Rogers, J.A. (2010), Improved Density in Aligned of Single-Walled

Hosseini, A. & Shabro V. (2010), Thermally-aware Modeling and Performance Evaluation

Hou, P.-X.; Liu, C. & Cheng, H.-M. (2008), Purification of Carbon Nanotubes, *Carbon*, Vol.

Hu, L.; Gruner, G.; Gong, J.; Kim, C.-J. "CJ" & Hornbostel, B. (2007), Electrowetting Devices

Design Hints based on Frequency- and Time-Domain Crosstalk Analyses, *IEEE*

*Characterization, Functionalization, and Applications*, Wiley-VCH, ISBN 978-3-527-

Nanotube Bundles for Interconnect Applications, *IEEE Electron Device Letters*, Vol.

Performance Projections for Ballistic Carbon-Nanotube Transistors, *IEEE*

based on C60 Fullerene Encapsulated Single-Walled Carbon Nanotubes, *Applied*

*Modeling, Integration and Applications*, Wiley-VCH, ISBN 978-3-527-31720-2, Federal

Carbon Nanotubes by Sequential Chemical Vapor Deposition on Quartz, *Advanced*

for Single-Walled Carbon Nanotube-based Interconnects for Future High Performance Integrated Circuits, *Microelectronic Engineering*, Vol. 87, No. 10, pp.

with Transparent Single-Walled Carbon Nanotube Electrodes, *Applied Physics*

Cambridge University Press, United States of America.

*Transactions on Electron Devices*, Vol. 58, No. 8, pp. 2702-2711.

ISBN 978-0-471-78046-5, United States of America.

*Bioanalytical Chemistry*, Vol. 384, No. 2, pp. 322-335.

*Transactions on Nanotechnology*, Vol. 5, No. 1, pp. 14-22.

Composites, *Current Applied Physics*, Vol. 7, No. 1, pp. 60-63.

*Physics Letters*, Vol. 97, No. 3, pp. 013104(3).

*Materials*, Vol. 22, No. 16, pp. 1826-1830.

32406-4, Federal Republic of Germany.

pp. 183-191.

United States of America.

28, No. 8, pp. 756-759.

4, United States of America.

Republic of Germany.

46, No. 15, pp. 2003-2025.

*Letters*, Vol. 90, No. 9, pp. 093124(3).

1955-1962.

1534.

Film-Si Schottky Contacts using Metal-Semiconductor-Metal Structures, *Applied Physics Letters*, Vol. 92, No. 24, pp. 243116(3).

Bradley, K.; Gabriel, J.-C.P.; Briman, M.; Star, A. and Grüner, G. (2003), Charge Transfer

Burghard, M.; Klauk, H. & Kern, K. (2009), Carbon-based Field-Effect Transistors for Nanoelectronics, *Advanced Materials*, Vol. 21, Nos. 25-26, pp. 2586-2600. Burke, P.J. (2004), AC Performance on Nanoelectronics: Towards a Ballistic THz Nanotube

Cao, Q.; Xia, M.-G.; Shim, M. & Rogers, J.A. (2006), Bilayer Organic-Inorganic Gate

Substrates, *Advanced Functional Materials*, Vol. 16, No. 18, pp. 2355-2362. Cao, Q.; Xia, M.; Kocabas, C.; Shim, M.; Rogers, J.A. & Rotkin, S.V. (2007), Gate Capacitance

Cao, Q. & Rogers, J.A. (2008), Random Networks and Aligned Arrays of Single-Walled

Cao, Q. & Rogers, J.A. (2009), Ultrathin Films of Single-Walled Carbon Nanotubes for

Chen, C. & Zhang, Y. (2009), *Nanowelded Carbon Nanotubes: From Field-Effect Transistors to*

Chen, P.-C.; Shen, G.; Sukcharoenchoke, S. & Zhou, C. (2009), Flexible and Transparent

Chen, P.-C.; Sukcharoenchoke, S.; Ryu, K.; de Arco, L.G.; Badmaev, A.; Wang, C. & Zhou, C.

Chen, X.; Wang, H.; Wan, H.; Song, K & Zhou, G. (2011), Semiconducting States and

Cho, H.; Koo, K.-H.; Kapur, P. & Saraswat, K. C. (2008), Performance Comparisons between

Danilchenko, B. A.; Shpinar, L.I.; Tripachko, N.A.; Voitsihovska, E.A.; Zelensky, S.E. &

Facchetti, A. & Marks, T.J. (2010), *Transparent Electronics: From Synthesis to Applications*, First

Edition, John Wiley & Sons, ISBN 978-0-470-99077-3, Great Britain.

Nanotube Bundles, *Applied Physics Letters*, Vol. 97, No. 7, pp. 072106(3). Dong, X.; Lau, C.M.; Lohani, A.; Mhaisalkar, S.G.; Kasim, J.; Shen, Z.; Ho, X.; Rogers, J.A. &

*Solar Microcells*, Springer, ISBN 978-3-642-01498-7, Germany.

*Applied Physics Letters*, Vol. 94, No. 4, pp. 043113(3).

*Condensed Matter*, Vol. 23, No. 31, pp. 315304(8).

*Electron Device Letters*, Vol. 29, No. 1, pp. 122-124.

*Materials*, Vol. 20, No. 12, pp. 2389-2393.

Transistor, *Solid-State Electronics*, Vol. 48, Nos. 10-11, pp. 1981-1986.

*Physics Letters*, Vol. 92, No. 24, pp. 243116(3).

*Letters*, Vol. 90, No. 2, pp. 023516(3).

*Materials*, Vol. 21, No. 1, pp. 29-53.

pp. 218301(4).

pp. 259-272.

pp. 1900-1904.

Film-Si Schottky Contacts using Metal-Semiconductor-Metal Structures, *Applied*

from Ammonia Physisorbed on Nanotubes, *Physical Review Letters*, Vol. 91, No. 21,

Dielectrics for High-Performance, Low-Voltage, Single-Walled Carbon Nanotube Thin-Film Transistors, Complementary Logic Gates, and p-n Diodes on Plastic

Coupling of Single-Walled Carbon Nanotube Thin-Film Transistors, *Applied Physics*

Carbon Nanotubes for Electronic Device Applications, *Nano Research*, Vol. 1, No. 4,

Electronics and Sensors: A Review of Fundamental and Applied Aspects, *Advanced*

Supercapacitor based on In2O3 Nanowire/Carbon Nanotube Heterogeneous Films,

(2010), 2,4,6-Trinitrotoluene (TNT) Chemical Sensing based on Aligned Single-Walled Carbon Nanotubes and ZnO Nanowires, *Advanced Materials*, Vol. 22, No. 17,

Transport in Metallic Armchair-Edged Graphene Nanoribbons, *Journal of Physics:*

Cu/Low-*κ*, Carbon-Nanotube, and Optics for Future on-Chip Interconnects, *IEEE*

Sundqvist, B. (2010), High Temperature Luttinger Liquid Conductivity in Carbon

Li, L.-J. (2008), Electrical Detection of Femtomolar DNA via Gold-Nanoparticle Enhancement in Carbon-Nanotube-Network Field-Effect Transistors, *Advanced*

Carbon Nanotube- and Graphene Based Devices, Circuits and Sensors for VLSI Design 63

Kocabas, C.; Kim, H.-S.; Banks, T.; Rogers, J.A.; Pesetski, A.A.; Baumgardner, J.E.;

Kreupl, F.; Graham, A.P.; Duesberg, G.S.; Steinhögl, W.; Liebau, M.; Unger, E. & Hönlein, W.

Krompiewski, S. (2005), Spin-Polarized Transport through Carbon Nanotubes, *Physica Status*

Law, M.; Goldberg, J. & Yang, P. (2004), Semiconductor Nanowires and Nanotubes, *Annual*

Lefenfeld, M.; Blanchet, G. & Rogers, J.A. (2003), High-Performance Contacts in Plastic

Lekakou, C.; Moudam, O.; Markoulidis, F.; Andrews, T.; Watts, J.F. & Reed, G. T. (2011),

Léonard, F. (2009), *The Physics of Carbon Nanotube Devices*, William Andrew, ISBN 978-0-

Li, H.; Yin, W.-Y.; Banerjee, K. & Mao, J.-F. (2008), Circuit Modeling and Performance

Li, H.; Xu, C.; Srivastava, N. & Banerjee, K. (2009), Carbon Nanomaterials for Next-

Li, J.; Ye, Q.; Cassell, A.; Ng, H.T.; Stevens, Ramsey; Han, Jie & Meyyappan, M. (2003),

Lin, A.; Patil, N.; Ryu, K.; Badmaev, A.; De Arco, L.G.; Zhou, C.; Mitra, S. & Wong, H.-S.P.

Nanotube FETs, *IEEE Transactions on Nanotechnology*, Vol. 8, No. 1, pp. 4-9. Lin, C.-T.; Hsu, C.-H.; Lee, C.-H. & Wu, W.-J. (2011), Inkjet-Printed Organic Field-Effect

Lin, Y.M.; Valdes-Garcia, A.; Han, S.-J.; Farmer, D.B.; Meric, I.; Sun, Y.; Wu, Y.;

Graphene Integrated Circuit, *Science*, Vol. 332, No. 6035, pp. 1294-1297. Liu, X.; Luo, Z.; Han, S.; Tang, T.; Zhang, D. & Zhou, C. (2005), Band Engineering of Carbon

*Transactions on Electron Devices*, Vol. 56, No. 9, pp. 1799-1821.

*Solidi B*: *Basic Solid State Physics*, Vol. 242, No. 2, pp. 226-233.

*Review of Materials Research*, Vol. 34, pp. 83-122.

*Nanotechnology*, Vol. 2011, Article ID 409382(8).

*Electron Devices*, Vol. 55, No. 6, pp. 1328-1337.

*Physics Letters*, Vol. 86, No. 24, pp. 243501(3).

8155-1573-9, United States of America.

Vol. 82, No. 15, pp. 1566791(3).

3206-3215.

1188-1191.

142890(7).

Vol. 64, Nos. 1-4, pp. 399-408.

Krishnaswamy, S.V. & Zhang, H. (2008), Radio Frequency Analog Electronics based on Carbon Nanotube Transistors, *PNAS*, Vol. 105, No. 5, pp. 1405-1409. Koo, K.-H.; Cho, H.; Kapur, P. & Saraswat, K.C. (2007), Performance Comparisons between

Carbon Nanotubes, Optical, and Cu for Future High-Performance On-Chip Interconnect Applications, *IEEE Transactions on Electron Devices*, Vol. 54, No. 12, pp.

(2002), Carbon Nanotubes in Interconnect Applications, *Microelectronic Engineering*,

Transistors and Logic Gates That Use Printed Electrodes of DNNSA-PANI Doped with Single-Walled Carbon Nanotubes, *Advanced Materials*, Vol. 15, No. 14, pp.

Carbon-based Fibrous EDLC Capacitors and Supercapacitors, *Journal of*

Analysis of Multi-Walled Carbon Nanotube Interconnects, *IEEE Transactions on*

Generation Interconnects and Passives: Physics, Status, and Prospects, *IEEE*

Bottom-Up Approach for Carbon Nanotube Interconnects, *Applied Physics Letters*,

(2009), Threshold Voltage and On-Off Ratio Tuning for Multiple-Tube Carbon

Transistor by Using Composite Semiconductor Material of Carbon Nanoparticles and Poly(3-Hexylthiophene), *Journal of Nanotechnology*, Vol. 2011, Article ID

Dimitrakopoulos, C.; Grill, A.; Avouris, P. & Jenkins, K.A. (2011), Wafer-Scale

Nanotube Field-Effect Transistors via Selected Area Chemical Gating, *Applied*

Hur, S.-H.; Khang, D.-Y.; Kocabas, C. & Rogers, J.A. (2004), Nanotransfer Printing by Use of

Ishikawa, F.N.; Stauffer, B.; Caron, D.A. & Zhou, C. (2009), Rapid and Label-Free Cell

Ishikawa, F.N.; Curreli, M.; Olson, C.A.; Liao, H.-I.; Sun, R.; Roberts, R.W.; Cote, R.J.;

Jain, D.; Rouhi, N.; Rutherglen, C.; Densmore, C. G.; Doorn, S. K. & Burke, P.J. (2011), Effect

Jamaa, M.H.B. (2011), *Regular Nanofabrics in Emerging Technologies: Design and Fabrication*

Javey, A. (2008), The 2008 Kavli Prize in Nanoscience: Carbon Nanotubes, *ACS Nano*, Vol. 2,

Javey, A. & Kong, J. (2009), *Carbon Nanotube Electronics*, Springer, ISBN 978-0-387-36833-7,

Jia, J.; Guan, W.; Sim, M.; Li, Y. & Li, H. (2008), Carbon Nanotubes based Glucose Needle-

Jia, Y.; Cao, A.; Bai, X.; Li, Z.; Zhang, L.; Guo, N.; Wei, J.; Wang, K.; Zhu, H.; Wu, D. &

Joachim, C.; Gimzewski, J.K. & Aviram, A. (2000), Electronics using Hybrid-Molecular and

Kang, S.J.; Kocabas, C.; Kim, H.-S.; Cao, Q.; Meitl, M.A.; Khang, D.-Y. & Rogers, J. A. (2007),

Kocabas, C.; Hur, S.-H.; Gaur, A.; Meitl, M.A.; Shim, M. & Rogers, J.A. (2005), Guided

Kocabas, C.; Shim, M. & Rogers, J.A. (2006), Spatially Selective Guided Growth of High-

for Electronic Applications, *Nano Letters*, Vol. 7, No. 11, pp. 3343-3348. Kanungo, M.; Malliaras, G.G. & Blanchet, G.B. (2010), High Performance Organic

Deficient Olefin, *Applied Physics Letters*, Vol. 97, No. 5, pp. 053304(3). Kim, G.; Bernholc, J. & Kwon, Y.-K. (2010), Band Gap Control of Small Bundles of Carbon

Mono-Molecular Devices, *Nature*, Vol. 408, No. 6812, pp. 541-548.

Ajayan, P.M. (2011), Achieving High Efficiency Silicon-Carbon Nanotube Heterojunction Solar Cells by Acid Doping, *Nano Letters*, Vol. 11, No. 5, pp. 1901-

Printed Multilayer Superstructures of Aligned Single-Walled Carbon Nanotubes

Transistors: Percolation Arrays of Nanotubes Functionalized with an Electron

Nanotubes using Applied Electric Fields: A Density Functional Theory Study,

Growth of Large-Scale, Horizontally Aligned Arrays of Single-Walled Carbon Nanotubes and Their Use in Thin-Film Transistors, *Small*, Vol. 1, No. 11, pp. 1110-

Coverage Arrays and Random Networks of Single-Walled Carbon Nanotubes and Their Integration into Electronic Devices, *Journal of the American Chemical Society*,

Arrays, *Journal of Nanomaterials*, Vol. 2011, Article ID 174268(7).

*Letters*, Vol. 85, No. 23, pp. 5730-5732.

States of America.

1905.

1116.

No. 7, pp. 1329-1335.

United States of America.

*Bioelectronics*, Vol. 24, No. 10, pp. 2967-2972.

Conditions, *ACS Nano*, Vol. 4, No. 11, pp. 6914-6922.

Type Biosensor, *Sensors*, Vol. 8, No. 3, pp. 1712-1718.

*Applied Physics Letters*, Vol. 97, No. 6, pp. 063113(3).

Vol. 128, No. 14, pp. 4540-4541.

Noncovalent Surface Forces: Applications to Thin-Film Transistors that Use Single-Walled Carbon Nanotube Networks and Semiconducting Polymers, *Applied Physics*

Detection by Metal-Cluster-decorated Carbon Nanotube Biosensors, *Biosensors and*

Thompson, M.E. & Zhou, C. (2010), Importance of Controlling Nanotube Density for Highly Sensitive and Reliable Biosensors Functional in Physiological

of Source, Surfactant, and Deposition Process on Electronic Properties of Nanotube

*Methods for Nanoscale Digital Circuits*, Springer, ISBN 978-94-007-0649-1, United

Carbon Nanotube- and Graphene Based Devices, Circuits and Sensors for VLSI Design 65

Rueckes, T.; Kim, K.; Joselevich, E.; Tseng, G.Y.; Cheung, C.-L. & Lieber, C.M. (2000) Carbon

Sangwan, V.K.; Behnam, A.; Ballarotto, V.W.; Fuhrer, M.S.; Ural, A. & Williams, E.D. (2010),

Schaman-Diamand, Y.; Osaka, T.; Datta, M. & Ohba, T. (2009), *Advanced Nanoscale ULSI*

Singh, D.V.; Jenkins, K.A.; Appenzeller, J.; Neumayer, D.; Grill, A. & Wong, H.-S. (2004),

Sinha, N.; Ma, J. & Yeow, J.T.W. (2006), Carbon Nanotube-based Sensors, *Journal of*

Srivastava, N. & Banerjee, K. (2004), Interconnect Challenges for Nanoscale Electronic

Srivastava, N.; Li, H.; Kreupl, F. & Banerjee, K. (2009), On the Applicability of Single-Walled

Star, A.; Han, T.-R.; Joshi, V.; & Stetter, J.R. (2004) Sensing with Nafion Coated Carbon Nanotube Field-Effect Transistors, *Electroanalysis*, Vol. 16, Nos. 1-2, pp. 108-112. Tan, C.S.; Gutmann, R.J. & Reif, L.R. (2008), *Wafer Level 3-D ICs Process Technology*, Springer,

Terrones, M. (2003), Science and Technology of the Twenty-First Century: Synthesis,

Terrones, M. (2004), Carbon Nanotubes: Synthesis and Properties, Electronic Devices and

Traversi, F.; Russo, V. & Sordan, R. (2009), Integrated Complementary Graphene Inverter,

Tseng, Y.-C.; Xuan, P.; Javey, A.; Malloy, R.; Wang, Q.; Bokor, J. & Dai, H. (2004), Monolithic

Tulevski, G.S.; Hannon, J.; Afzali, A.; Chen, Z.; Avouris, P. & Kagan, C.R. (2007), Chemically

Vaillancourt, J.; Zhang, H.; Vasinajindakaw, P.; Xia, H.; Lu, X.; Han, X.; Janzen, D.C.; Shin,

Vasileska, D. & Goodnick, S.M. (2011), *Nano-Electronic Devices: Semiclassical and Quantum Transport Modeling*, Springer, ISBN 978-1-4419-8839-3, United States of America.

Circuits, *TMS Journal of Materials (JOM)*, Vol. 56, No. 10, pp. 30-31.

*Applied Physics Letters*, Vol. 97, No. 4, pp. 043111(3).

*Transactions on Nanotechnology*, Vol. 3, No. 3, pp. 383-387.

*Nanoscience and Nanotechnology*, Vol. 6, No. 3, pp. 573-590.

ISBN 978-0-387-76532-7, United States of America.

*Applied Physics Letters*, Vol. 34, No. 22, pp. 223312(3).

*Applied Physics Letters*, Vol. 93, No. 24, pp. 243301(3).

*Science*, Vol. 289, pp. 94-97.

United States of America.

8, No. 4, pp. 542-559.

325-377.

11964-11968.

*Research*, Vol. 33, pp. 419-501.

*Letters*, Vol. 4, No. 1, pp. 123-127.

Nanotube-based Nonvolatile Random Access Memory for Molecular Computing,

Optimizing Transistor Performance of Percolating Carbon Nanotube Networks,

*Interconnects: Fundamentals and Applications*, Springer, ISBN 978-0-387-95867-5,

Frequency Response of Top-Gated Carbon Nanotube Field-Effect Transistors, *IEEE*

Carbon Nanotubes as VLSI Interconnects, *IEEE Transactions on Nanotechnology*, Vol.

Properties, and Applications of Carbon Nanotubes, *Annual Review of Materials*

Other Emerging Applications, *International Materials Reviews*, Vol. 49, No. 6, pp.

Integration of Carbon Nanotube Devices with Silicon MOS Technology, *Nano*

Assisted Directed Assembly of Carbon Nanotubes for the Fabrication of Large-Scale Device Arrays, *Journal of the American Chemical Society*, Vol. 129, No. 39, pp.

W.-S.; Jones, C.S.; Stroder, M.; Chen, M.Y.; Subbaraman, H.; Chen, R.T.; Berger, U. & Renn, M. (2008), All Ink-Jet-Printed Carbon Nanotube Thin-Film Transistor on a Polyimide Substrate with an Ultrahigh Operating Frequency of Over 5 GHz,

Lu, W. & Lieber, C.M. (2007), Nanoelectronics from the Bottom Up, *Nature Materials*, Vol. 6,

Mahar, B.; Laslau, C.; Yip, R. & Sun, Y. (2007), Development of Carbon Nanotube-based

Mallick, G.; Griep, M.H.; Ayajan, P.M. & Karna, S.P. (2010), Alternating Current-to-Direct

Mamalis, A.G.; Vogtländer, L.O.G. & Markopoulos, A. (2004), Nanotechnology and

Nismi, N.A.; Adikaari, A.A.D.T. & Silva, S.R.P. (2010), Functionalized Multiwall Carbon

Nogueira, A. F.; Lomba, B.S.; Soto-Oviedo, M.A.; Correia, C.R.D.; Corio, P.; Furtado, C.A. &

*Nanomaterials, Interfaces and Hard Matter*, Vol. 111, No. 49, pp. 18431-18438. Nojeh, A. & Ivanov, A. (2010), Wireless Interconnect and the Potential for Carbon Nanotubes, *IEEE Design & Test of Computers*, Vol. 27, No. 4, pp. 44-52. Nouchi, R.; Tomita, H.; Ogura, A.; Shiraishi, M. & Kataura, H. (2008), Logic Circuits using

Novoselov, K.S.; Geim, A.K.; Morozov, S.V.; Jiang, D.; Zhang, Y.; Dubonos, S.V.; Grigorieva,

Oliveira, A.C. & Mascaro, L.H. (2011), Evaluation of Acetylcholinesterase Biosensor based

Papanikolaou, A.; Soudris, D. & Radojcic, R. (2011), *Three Dimensional System Integration: IC*

Pesetski, A.A.; Baumgardner, J. E.; Krishnaswamy, S.V.; Zhang, H.; Adam, J. D.; Kocabas, C.;

Philip-Wong, H.-S. & Akinwande, D. (2011), *Carbon Nanotube and Graphene Device Physics*, Cambridge University Press, ISBN 978-0-521-51905-2, United Kingdom. Reddy, D.; Register, L.F.; Carpenter, G.D. & Banerjee, S.K. (2011), Graphene Field-Effect Transistors, *Journal of Physics D: Applied Physics*, Vol. 44, No. 31, pp. 313001(20). Rivas, G.A.; Rubianes, M.D.; Pedano, M.L.; Ferreyra, N.F.; Luque, G. & Miscoria, S.A. (2009),

Rowell, M.W.; Topinka, M.A.; McGehee, M.D.; Prall, H.-J.; Dennler, G.; Sariciftci, N.S.; Hu,

*Journal of Analytical Chemistry*, Vol. 2011, Article ID 974216 (6).

Publishers, ISBN 978-1-607-41314-1, United States of America.

Electrodes, *Applied Physics Letters*, Vol. 88, No. 23, pp. 233506(3).

*Applied Physics Letters*, Vol. 93, No. 12, pp. 123506(2).

Current Power Conversion by Single-Wall Carbon Nanotube Diodes, *Applied*

Nanostructured Materials: Trends in Carbon Nanotubes, *Precision Engineering*, Vol.

Nanotubes incorporated Polymer/Fullerene Hybrid Photovoltaics, *Applied Physics*

Hümmelgen, I.A. (2007), Polymer Solar Cells using Single-Wall Carbon Nanotubes Modified with Thiophene Pedant Groups, *Journal of Physical Chemistry C:*

Solution-Processed Single-Walled Carbon Nanotube Transistors", *Applied Physics*

I.V. & Firsov, A.A. (2004), Electric Field Effect in Atomically Thin Carbon Films,

on Carbon Nanotube Paste in the Determination of Chlorphenvinphos, *International*

*Stacking Process and Design*, Springer, ISBN 978-1-4419-0961-9, United States of

Banks, T. & Rogers, J.A. (2008), A 500 MHz Carbon Nanotube Transistor Oscillator,

*Carbon Nanotubes: A New Alternative for Electrochemical Sensors*, Nova Science

L. & Gruner, G. (2006), Organic Solar Cells with Carbon Nanotube Network

Sensors – A Review, *IEEE Sensors Journal*, Vol. 7, No. 2, pp. 266-284.

Marulanda, J.M. (2010), *Carbon Nanotubes*, In-Tech, ISBN 978-953-307-054-4, Croatia.

*Physics Letters*, Vol. 96, No. 23, pp. 233109(3).

*Letters*, Vol. 97, No. 3, pp. 033105(3).

*Letters*, Vol. 92, No. 25, pp. 253507(3).

*Science*, Vol. 306, No. 5696, pp. 666-669.

No. 11, pp. 841-850.

28, No. 1, pp. 16-30.

America.

**1. Introduction**

2004), (Brooks D., 2003).

mathematical model is presented.

**0**

**4**

*Mexico*

**Impedance Matching in VLSI Systems**

The continuous scaling process into submicrometric dimensions of silicon based devices has allowed the integration of a large number of systems in a single chip. Besides, the operating frequencies of such systems are higher and a large amount of information can be processed in a short period of time. On the other hand, while the core frequencies are increasing, higher data rates for off-chip interconnections become necessary, for example a processor that communicates with the memory in order to process information. Unfortunately, at high rates the signal wave length is comparable with the physical length of the interconnections, because of this, parasitic and transmission line effects have to be taken into account. As a consequence, the transmitted signal integrity is degraded resulting in communication errors (Thierauf S.,

It has been shown that, for modern off-chip communication systems, current mode signaling offers several advantages over voltage-mode at high data rates (Juan, 2007), but they need to be matched in impedance to the interconnection line. However, impedance matching requires termination resistors. Moreover, due to the large number of input/output circuits in a single chip, terminations have to be placed on-chip (Fan Y. & Smith J., 2003) so that the PCB area is not increased. One of the most important transmission line effects that degrades signal integrity in these signaling schemes is reflection loss. In this case, signal reflections traveling trough the line are present in either driver to receiver or receiver to driver directions. Unfortunately, it is difficult to achieve perfect matching of impedances due to the large process variations in the fabrication of interconnection lines and the different traces between them (Ramachandran N. et. al., 2003). Also, temperature variations and external effects are present inside and outside the chip. As a conclusion, impedance matching techniques must

be developed in order to automatically adapt the impedance variations of the line.

In this chapter systems for on-die automatic impedance matching for off-chip signaling are described. In order to perform the automatic matching operation an algorithm that integrates the sign of the impedance matching error and the sign of the coupling branch current is implemented. The advantage of this algorithm is that it works without interfering with the driver operation. Computer simulations of layout extractions are presented. Also, a system of knowlegde- based impedance matching which avoids the calculation of a complex

Díaz Méndez J. Alejandro1, López Delgadillo Edgar2

<sup>1</sup>*National Institute for Astrophysics, Optics and Electronics*

and Arroyo Huerta J. Erasmo1

<sup>2</sup>*Universidad Autónoma de Aguascalientes*


**5. Discussion**

Japanese encephalitis is a neuroinvasive virus, which is destructive of neural tissue. Loss of neurovirulence is an important consideration in live JE vaccine development. It is well known that the mice and Rhesus monkeys are highly sensitive animals for testing neurovir‐ ulence of JE virus. Both were used during development of live JE vaccine. The process of derivation of the SA14-14-2 strain has demonstrated fine balances between stable neuroat‐ tenuation and immunogenicity.

The principal issue surrounding the clinical application of the SA14-14-2 live virus vaccine candidate is safety, especially absence of neurovirulence and stable even passage in brain tissue. During development of the SA14-14-2 strain, it was demonstrated that unstable attenu‐ ated virus clones i.e. SA14-12-1-7 and SA14-12-1-1 could be detected by several passages in PHK cells or one i.c. mouse passage. Using this method, SA14-14-2 vaccine virus was select‐ ed for its neuroattenuation stability. To ensure vaccine safety, absence of neuroreversion after one ic passage in suckling mice is required in the quality control for vaccine production.

Compared with other live attenuated viral strains, SA14-14-2 has the following characteristics:

trols. Therefore, IL-6, IL-8, MCP-1,MCP-1α and MIP-1β may play important roles in the im‐

Besides Xu et al.[51] reported that mice immunized with SA14-14-2 virus non-structural NS1 protein, which expressed and purified from SA14-14-2 virus NS1 recombinant *E.coli* BL21 (DE3), were protected against a lethal JEV challenge (50-70% protection). Guinea-pigs vacci‐ nated with the NS1 protein presented with reduced viremia following challenge with viru‐

SA14-14-2 NS1 1 0a 0 0 0 0 0 0

Unimmunized controls 1 0 7. 5 > 579 208 0 0 0

**Table 18.** Viremia suppression in guinea pigs vaccinated with SA14-14-2 NS1 protein followed by challenge with virulent JEV. ND, Not determinedGuinea pigs were intraperitoneally (i.p.) vaccinated with NS1 protein, each 60 ug, at day 0 and 7 respectively. Fourteen days after the first vaccination, each vaccinated and unvaccinated animal was i.p.

Japanese encephalitis is a neuroinvasive virus, which is destructive of neural tissue. Loss of neurovirulence is an important consideration in live JE vaccine development. It is well known that the mice and Rhesus monkeys are highly sensitive animals for testing neurovir‐ ulence of JE virus. Both were used during development of live JE vaccine. The process of derivation of the SA14-14-2 strain has demonstrated fine balances between stable neuroat‐

The principal issue surrounding the clinical application of the SA14-14-2 live virus vaccine candidate is safety, especially absence of neurovirulence and stable even passage in brain tissue. During development of the SA14-14-2 strain, it was demonstrated that unstable attenu‐ ated virus clones i.e. SA14-12-1-7 and SA14-12-1-1 could be detected by several passages in PHK cells or one i.c. mouse passage. Using this method, SA14-14-2 vaccine virus was select‐ ed for its neuroattenuation stability. To ensure vaccine safety, absence of neuroreversion after one ic passage in suckling mice is required in the quality control for vaccine production.

challenged with virulent JEV P3 strain (each 2.5ml).a No virus detected in the undiluted serum

0 1 3 5 7 10 14

 0 0 0 0 0 0 0 0 0 2. 5 0 0 0 0 0 7. 5 20 0 0 0 0 0 17. 5 145 15 0 0 0

 0 47. 5 260 0 0 0 0 0 22. 5 318 230 2. 5 0 0 0 7. 5 225 0 0 0 0 0 0 > 750 ND ND ND ND

mune response to JE live attenuated vaccine in the humans.

**Tested groups No. animals Viremia (pfu/ml) by days postchallenge**

lent JE virus (Table 18)

198 Encephalitis

**5. Discussion**

tenuation and immunogenicity.

As for the immunogenicity of the SA14-14-2 vaccine, studies have demonstrated that protec‐ tion efficacy of the live attenuated vaccine was mediated by the presence of neutralizing an‐ tibodies and by potent cell-mediated immunity as well as the NS1 protein induced immunity, which associated with the high efficacy of the SA14-14-2 vaccine. Besides anam‐ nestic immune response will give quick rise of neutralizing antibody and provide long-term protection against JE infection among subjects who become seronegative after immunization with SA14-14-2 live vaccine [39]

Live JE vaccine (SA14-14-2) has been used in more than 300 million children since large scale production began in 1989. To date no vaccine associated JE cases has been reported in China and outside.The safety of live JE vaccine is due to a high degree of neuroattenuation and a number of stable phenotypic and genotypic characteristics. The combination of lack of vire‐ mia in the vaccinees and the absence of virus replication and dissemination in the mosquito vector make the likelihood of transmission of JE vaccine virus and the risk of environment highly unlikely. Therefore the exceptional safety, stability, immunogenicity and long-term protective efficacy present a strong case for the expended use of this live attenuated JE vac‐ cine in the world.

[8] Fan, X. L., Yu, Y. X., Li, D. F., & Yao, Z. H. (2002a). Comparison of nucleotide and deduced amino acid sequences of E protein gene of the wild-type Japanese encepha‐ litis virus strain SA14 and its different attenuated derivatives. *Virologica Sinica*, 17,

Development of Japanese Encephalitis Attenuated Live Vaccine Virus SA14-14-2 and its Charcteristics

http://dx.doi.org/10.5772/ 52980

201

[9] Fan, X. L., Yu, Y. X., Li, D. F., & Yao, Z. H. (2002b). The stability of E protein gene of the Japanese encephalitis live-attenuated vaccine virus SA14-14-2. *Chin J Virology*, 18,

[10] Feng, Y., Zhang, H. L., Yu, Y. X., Zhang, Z. Y., Yang, W. H., Dong, G. M., et al. (2007). Observations on the replication of virus and stability of virulence in Culex tritaenio‐ rhynchus after intrathoracically inoculation with attenuated Japanese encephalitis vi‐

[11] Gao, X. J., Zhao, Y. J., Liu, C. M., Zou, Y., Wei, Y., Zheng, X. G., et al. (2006). The sta‐ bility of E gene of the Japanese encephalitis attenuated virus strain SA14-14-2. *Prog in*

[12] Gatchalian, S., Yao, Y., Zhou, B., et al. Comparison of the immunogenicity and safety of measles vaccine administered alone or with live, attenuated Japanese encephalitis

[13] Guirakhoo, F., Zhang, Z. X., Chambers, T. J., Delagrave, S., Arroyo, J., Barrett, A. D. T., et al. (1999). Immunogenicity, genetic stability, and protective efficacy of a re‐ combinant, chimeric yellow fever-Japanese encephalitis virus (ChimeriVax-JE) as a live, attenuated vaccine candidate against Japanese encephalitis. *Virology*, 257,

[14] Hennessy, S., Liu, Z. L., Tsai, T. F., et al. (1996). Effectiveness of live-attenuated Japa‐ nese encephalitis vaccine (SA14-14-2): a case control study. *Lancet*, 347, 1583-87.

[15] Huang, Z. L., Li, X. G., & Wu, J. N. (1998). Observation on the side reaction after vac‐ cination with Japanese encephalitis live vaccine. *Chin J Zoonoses*, 14, 60-61.

[16] Jia, L. L., Zheng, Z., & Yu, Y. X. (1992). Study on the immune mechanism of JE atte‐ nuated live vaccine(SA14-14-2 strain). *Chin J Microbio Immuno*, 12, 364-366.

[17] Jia, L. L., Zheng, Z., Guo, Y. P., & Yu, Y. X. (1992). Stability of live-attenuated Japa‐

[18] Jia, L. L., Zheng, Z., & Yu, Y. X. (1993). Pathogenicity and Immunogenicity of attenu‐ ated Japanese encephalitis vaccine (SA14-14-2) in immune-inhibited mice. *Virologica*

[19] Jia, L. L., Zheng, Z., Wang, Z. W., et al. (1995a). Protective effect and antibody re‐ sponse in guinea-pigs immunized with Japanese encephalitis attenuated live vaccine

nese encephalitis vaccine strain SA14-14-2. *Chin J Biologicals*, 5, 174-6.

after challenge with virulent virus. *Prog Microbiol Immunol*, 23, 73-76.

SA14-14-2 vaccine in Philippine infants [J]. *Vaccine*, 26(18), 2234-2241.

rus SA14-14-2 vaccine strain. *Chin J Zoonoses*, 23, 278-281.

216-220.

39-47.

363-372.

*Sinica*, 8, 20-4.

*Microbiol Immunol*, 34, 1-11.


**2. Results**

*2.1. PE*/*HDP Patient Sera Up-Regulated Gene Expression of IL-6, CCL2, High Mobility Group Box (HMGB)1, S100 Ca2*+*-Binding Protein B (S100B), and Receptor for Advanced Glycation Endproducts (RAGE) in Primary Cultured Human Adipocytes*

Obesity increases PE/HDP risk. Maternal obesity, increased insulin resistance, and inflammation are involved in the pathogenesis of PE/HDP [24,25]. Furthermore, PE/HDP risk has been reported to increase 2–4-fold among women with diabetes [26]. We therefore hypothesized that the PE/HDP patient sera contain some of these factors that induce insulin resistance and/or inflammation. We incubated primary cultured human adipocytes with sera from disease-free pregnant women (control) or those from PE/HDP (patients) for 24 h, and the gene expression of *IL-6*, *CCL2*, *tumor necrosis factor* α (*TNF*α), *leptin* (*LEP*), *adiponectin* (*ADIP*), *resistin* (*RETN*), *HMGB1*, *S100B*, and *RAGE* in the adipocytes was measured via real-time reverse transcriptase-polymerase chain reaction (RT-PCR). As shown in Figure 1, mRNA levels of *IL-6*, *CCL2*, *HMGB1*, *S100B*, and *RAGE*, but not *TNF*α, *LEP*, *ADIP*, and *RETN* (*P* = 0.4496, *P* = 0.1157, *P* = 0.0875, and *P* = 0.2912, respectively) were significantly increased by the addition of PE/HDP patient sera compared to those cells incubated with control sera.

#### *2.2. Up-Regulation of IL-6 and CCL2 by HMGB1 and Advanced Glycation Endproducts (AGE) in Adipocytes*

It is well-known that HMGB1 and S100B are typical ligands for RAGE. RAGE expression was reported in adipocytes and SW872 cells [27,28], and furthermore immunofluorescent staining of RAGE in 3T3-L1 adipocytes was shown [27]. RAGE expression was up-regulated by ligands for RAGE [29], we tested whether ligands for RAGE up-regulate gene expression of inflammatory mediators, such as *IL-6* and *CCL2*, in human SW872 adipocytes. We added HMGB1, AGE, and S100B in SW872 culture medium, incubated for 24 h, and the expression of *IL-6* and *CCL2* was analyzed via real-time RT-PCR. As shown in Figure 2, mRNAs of *IL-6* and *CCL2* were significantly up-regulated by the addition of HMGB1 and AGE. In contrast, S100B, another noted ligand for RAGE, failed to up-regulate mRNA for *IL-6* or *CCL2*.

In order to see whether the up-regulation of mRNAs for *IL-6* and *Ccl2* occurred only in SW872 or other adipocytes, we cultured mouse 3T3-L1 preadipocytes, di fferentiated them into di fferentiated adipocytes, and tested whether ligands for RAGE up-regulate gene expression of *IL-6* and *Ccl2* in mouse 3T3-L1 undi fferentiated preadipocytes and di fferentiated adipocytes. As shown in Figure 3, the mRNA levels of *IL-6* were significantly up-regulated by AGE and HMGB1 but not by S100B (*P* = 0.6414) in di fferentiated 3T3-L1 adipocytes, but unchanged by any of the RAGE ligands (AGE, HMGB1, or S100B) in undi fferentiated preadipocytes ( *P* = 0.8037 [No addition vs. AGE], *P* = 0.4793 [No addition vs. HMGB1], and *P* = 0.3138 [No addition vs. S100B]). In contrast, the mRNA levels of *Ccl2* remained unchanged in response to AGE, HMGB1, or S100B in 3T3-L1 di fferentiated adipocytes

(*P* = 0.1892 [No addition vs. AGE], *P* = 0.2885 [No addition vs. HMGB1], and *P* = 0.4024 [No addition vs. S100B]), but significantly up-regulated in the undifferentiated preadipocytes by the addition of AGE but not by HMGB1 and S100B (*P* = 0.1241 [No addition vs. HMGB1] and *P* = 0.4305 [No addition vs. S100B]) (Figure 3). Previous studies reported that S100B up-regulated TNFα in adipocytes [30,31]. In contrast, S100B induced neither *IL-6* nor *CCL2* in adipocytes in this study, suggesting that SW872 and 3T3-L1 cells may insensitive to S100B.

**Figure 1.** The mRNA levels of *IL-6*, *CCL2*, *TNF*α, *LEP*, *ADIP*, *RETN*, *HMGB1*, *S100B*, and *RAGE* in primary cultured human adipocytes treated with sera from disease-free control (Control) or preeclampsia/hypertensive disorders of pregnancy (PE/HDP) patients (Patients) for 24 h. The levels of the mRNAs were measured via real-time reverse transcriptase-polymerase chain reaction (RT-PCR) using β*-actin* as an endogenous control. Data are expressed as mean ± SE for each group (*n* = 4). The statistical analyses were performed using Student's *t*-test.

**Figure 2.** The mRNA levels of *IL-6* and *CCL2* in SW872 human adipocytes treated with 1 μg/mL HMGB1, 150 μg/mL advanced glycation endproducts (AGE), or 100 ng/mL S100B for 24 h. The levels of the mRNAs were measured via real-time RT-PCR using β*-actin* as an endogenous control. Data are expressed as mean ± SE for each group (*n* = 4). The statistical analyses were performed using Student's *t*-test vs. No addition.

**Figure 3.** The mRNA levels of *IL-6* and *Ccl2* in 3T3-L1 mouse cells (undifferentiated preadipocytes and differentiated adipocytes) treated with 300 μg/mL AGE, 1 μg/mL HMGB1, or 100 ng/mL S100B for 24 h. The levels of the mRNAs were measured via real-time RT-PCR using *rat insulinoma gene* (*Rig*)/*ribosomal protein S15* (*RpS15*) as an endogenous control. Data are expressed as mean ± SE for each group (*n* = 4). The statistical analyses were performed using Student's *t*-test vs. No addition.

#### *2.3. Down-Regulation of RAGE Attenuated the Increases of IL-6 and CCL2 in Adipocytes Treated with Small Interfering RNA (siRNA) for RAGE*

In order to see the mechanism of HMGB1- and AGE-induced gene expression of *IL-6* and *CCL2*, *RAGE* gene was knocked down by RNA interference. The expression of *IL-6* and *CCL2* was significantly increased by the addition of HMGB1 and AGE even in the presence of scrambled RNA. In contrast, introduction of small interfering RNA (siRNA) for RAGE (*siRAGE*) clearly inhibited the HMGB1- and AGE-induced increases of mRNAs for *IL-6* and *CCL2* in SW872 human adipocytes (Figure 4; *P* = 0.2638 [No addition vs. HMGB1 in *IL-6*], *P* = 0.0744 [No addition vs. AGE in *IL-6*], *P* = 0.2559 [No addition vs. HMGB1 in *CCL2*], and *P* = 0.5541 [No addition vs. AGE in *CCL2*]).

We also measured the concentrations of IL-6 and CCL2 in the RAGE-knocked-down SW872 cell culture medium via enzyme-linked immunosorbent assay (ELISA). The concentrations of IL-6 and CCL2 were significantly increased in response to HMGB1 and AGE in scrambled RNA-introduced cell culture medium. In contrast, the introduction of siRAGE significantly attenuated the HMGB1- and AGE-induced increases of IL-6 and CCL2 in the medium (Figure 5).

#### *2.4. Up-Regulation of IL-6 and CCL2 by Lipopolysaccharide (LPS) in Adipocytes*

Recent reports indicated that PE/HDP is also induced by lipopolysaccharide (LPS) [32] and that RAGE mediates LPS signaling and acts as an LPS receptor [33–38]. Thus, we tested whether LPS up-regulate gene expression of *IL-6* and *CCL2* in human SW872 adipocytes. We added 10 ng/mL LPS in SW872 culture medium, incubated for 24 h, and the expression of *IL-6* and *CCL2* was analyzed via real-time RT-PCR. As shown in Figure 6, mRNAs of *IL-6* and *CCL2* were significantly up-regulated by the addition of LPS.

**Figure 4.** Effects of siRNA against *RAGE* on HMGB1- and AGE-induced gene expression of *IL-6* and *CCL2*. SiRNA for *RAGE* was transfected into SW872 cells and the cells were incubated with HMGB1 or AGE for 24 h. The levels of *IL-6* and *CCL2* mRNA were measured via real-time RT-PCR using β*-actin* as an endogenous control. Data are expressed as mean ± SE for each group (*n* = 4). The statistical analyses were performed using Student's *t*-test vs. No addition.

**Figure 5.** Effect of siRNA against RAGE on the HMGB1- and AGE-induced expression of IL-6 and CCL2. SiRNA for RAGE was transfected into SW872 cells and the cells were incubated with HMGB1 or AGE for 24 h. The levels of IL-6 and CCL2 in the cell culture medium were measured via ELISA. Data are expressed as mean ± SE for each group (*n* = 4). The statistical analyses were performed using Student's *t*-test vs. No addition.

**Figure 6.** The mRNA levels of IL-6 and CCL2 in SW872 human adipocytes treated with 10 ng/mL lipopolysaccharide (LPS) for 24 h. The levels of the mRNAs were measured via real-time RT-PCR using β-actin as an endogenous control. Data are expressed as mean ± SE for each group (*n* = 4). The statistical analyses were performed using Student's *t*-test.

We next measured IL-6 and CCL2 in the LPS-stimulated SW872 cell culture medium and found that the levels of IL-6 and CCL2 in the LPS-stimulated SW872 culture medium were also elevated significantly (Figure 7).

**Figure 7.** The levels of IL-6 and CCL2 in culture medium of SW872 human adipocytes treated with 10 ng/mL LPS for 24 h. The levels of IL-6 and CCL2 in the cell culture medium were measured via ELISA. Data are expressed as mean ± SE for each group (n = 4). The statistical analyses were performed using Student's *t*-test.

#### *2.5. Down-Regulation of RAGE Attenuated the LPS-Induced IL-6 and CCL2 Increases in Adipocytes*

In order to confirm whether the mechanism of LPS-induced *IL-6* and *CCL2* up-regulation is also mediated by RAGE, *RAGE* gene was knocked down by RNA interference. The expression of *IL-6* and *CCL2* was significantly increased by the addition of LPS even in the presence of scrambled RNA. In contrast, introduction of *siRAGE* clearly inhibited the LPS-induced increases of mRNAs for *IL-6* and *CCL2* in SW872 human adipocytes (Figure 8).

We also measured the concentrations of IL-6 and CCL2 in the RAGE-knocked-down SW872 cell culture medium via ELISA. The concentrations of IL-6 and CCL2 were significantly increased in response to the addition of LPS in scrambled RNA-introduced cell culture medium. In contrast, the introduction of siRAGE significantly attenuated the LPS-induced increases of IL-6 and CCL2 in the medium (Figure 9).

**Figure 8.** Effects of siRNA against *RAGE* on the LPS-induced gene expression of *IL-6* and *CCL2*. SiRNA for *RAGE* was transfected into SW872 cells and the cells were incubated with 10 ng/mL LPS for 24 h. The levels of *IL-6* and *CCL2* mRNA were measured via real-time RT-PCR using β*-actin* as an endogenous control. Data are expressed as mean ± SE for each group (*n* = 4). The statistical analyses were performed using Student's *t*-test.

**Figure 9.** Effect of siRNA against RAGE on the LPS-induced expression of IL-6 and CCL2. SiRNA for RAGE was transfected into SW872 cells and the cells were incubated with LPS for 24 h. The levels of IL-6 and CCL2 in the cell culture medium were measured via ELISA. Data are expressed as mean ± SE for each group (*n* = 4). The statistical analyses were performed using Student's *t*-test.


*3.2. Total Emissions of Gaseous Combustion Products*

The total emissions of the analyzed exhaust gas components were determined from the dependence (12) based on the above-mentioned measurement results and the adopted assumptions. The geometrical interpretation of the total emission of exhaust gas components is expressed by the fields under the graphs defining the emission stream of these components (see Figures 5–8). The obtained results are presented in Figure 9.

**Figure 9.** Curves describing changes in total emission of CO2, CO, NOx, and SO2 as a function of the R parameter. Dashed lines—extrapolated trend lines beyond the test range (value 0.1 of the R parameter).

The dashed lines show the trend lines extrapolated beyond the test range (by the value of 0.1 of the parameter R).


**4. Spatial estimation of soil erosion**

### **4.1 The R factor**

R factor determines erosivity by rainfall at a specific region, which is usually based on the intensity and amount of precipitation. It mainly represents the impact caused by rainfall intensity on soil erosion. In this study area, the monthly precipitation data for six weather stations situated in the study area were obtained from the World Bank website. It was validated with the satellite based data to integrate the results. Rainfall map was prepared which represents the spatial distribution of rain in the Eastern Hindukush. This map was utilized to estimate R factor map by calculating the Rainfall erosivity for a time period of about 1991 to 2019 in study area. Rainfall-runoff erosivity was calculated by the following equation used by many researchers on areas where similar topographic and atmospheric conditions prevail (**Figure 4**) [19, 22].

$$\mathbf{R} = \mathbf{0}.\mathbf{0}5 \times \mathbf{P}.\tag{14}$$

**Figure 4.** *Rainfall-Erosivity factor (R) of eastern Hindukush, Pakistan.*

Where, R = Rainfall Erosivity Factor, P = Mean Annual Rainfall in (mm).


**4. Conclusions**

In conclusion, cationic lignocellulose nanofibers (CLCNF) with nanoscale diameters and quaternary ammonium group contents in the range of 0.97–1.76 mmol/g were successfully prepared by cationic modification of bagasse in deep eutectic solvents (DES) followed by mechanical defibrillation. Such lignocellulosic nanomaterials showed excellent adsorption performance in removing dissolved and colloidal substances (DCS) model contaminant polygalacturonic acid (PGA). The cationic modification in DES changed the surface charge of bagasse from negative to positive and obtained a large number of active adsorption sites, so CLCNF can easily adsorb the negatively charged PGA in water. Electrostatic interactions were considered as the main mechanism for capturing PGA by CLCNF in water. The kinetic process of PGA adsorption by CLCNF can be predicted by a pseudo-second-order model and the Langmuir model fitted the data well, indicating monolayer adsorption. The CLCNF adsorbent in current research has the advantages of low cost, high efficiency and large adsorption capacity, which can meet the new demand of DCS adsorbents. It is expected to be used for the adsorption of other anionic contaminants in water.

**Supplementary Materials:** The following supporting information can be downloaded at: https: //www.mdpi.com/article/10.3390/polym14050910/s1, Figure S1: Optical microscope images of bagasse (A), the precursors of LCNF, CLCNF-1, CLCNF-2, and CLCNF-3 (B–E); Figure S2: Separation factor of PGA adsorption by CLCNF-3.

**Author Contributions:** Methodology, L.Y.; formal analysis, L.Y., X.L. and Y.J.; investigation, X.Z., L.Y., Y.J., H.Z. and G.C.; data creation, X.Z. and S.Z.; writing original draft preparation, L.Y.; writing review and editing, X.L. and Y.J.; supervision, X.L.; project administration, S.W. All authors have read and agreed to the published version of the manuscript.

**Funding:** The research was supported by the China Postdoctoral Science Foundation (2020M683209), the Natural Science Foundation of Guangxi (2020GXNSFBA159009, 2021GXNSFBA075043, 2018GXNS-FAA138126), the Opening Project of Guangxi Key Laboratory of Clean Pulp & Papermaking and Pollution Control (2019ZR04, 2019KF28), the Scientific Research Foundation of Guangxi University for Nationalities (2019KJQD10), Guangxi Ba-Gui Scholars Program, the Guangxi Major Projects of Science and Technology (GXMPSTAB21196064) and the Specific Research Project of Guangxi for Research Bases and Talents (AD18126002).

**Institutional Review Board Statement:** Not applicable.

**Informed Consent Statement:** Not applicable.

**Data Availability Statement:** Not applicable.

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