Contents

Preface

Animals have been used in biological and medical research for thousands of years in order to advance the comprehension of living beings and to study the origin and basic principles of life. Furthermore, animal models have contributed significantly to a better understanding of general anatomy, physiology, pathology and pharmacology, as they can be utilized to create controlled situations and simulate specific

Without a doubt, animal models have played essential roles in the most important advances in numerous medical and biological fields. From Louis Pasteur's demonstration of the germ theory of disease using anthrax in sheep and Thomas Hunt Morgan's work with the fruit fly *Drosophila melanogaster* identifying chromosomes as the vector of inheritance for genes, to Jonas Salk's studies on rhesus monkeys to isolate the polio virus, which led to the creation of a polio vaccine and modern medical advances such as organ transplant techniques and the heart–lung machine, animal models have significantly contributed to improvement in the quality of human life. New drugs, vaccines and treatment protocols for human and animal diseases have also been developed thanks to diverse animal models. The progress achieved thanks to the creation and use of animal models is indubitable, as the majority of Nobel Prize awardees in Physiology and Medicine used animal experi-

This book is a collection of 13 state-of-the-art chapters written by renowned experts from all over the world. It represents a compilation of scientific and clinical data that emphasize findings in a wide variety of specialties, such as neurology, psychiatry, cardiology, and behavioral data including the eyes and ears, pediatrics, skin disorders, cardiovascular and musculoskeletal systems, reproduction, chronic

The book provides basic, up-to-date background on the topics as well as deals with clinical aspects, providing insightful information related to the use of animal models in the understanding of the pathophysiological mechanisms of action of

A critical objective of the book is to provide real-time experimental approaches and data to scientists, clinicians, researchers and students for the identification of new therapeutic targets and biomarkers using animal models as well as investigate the

We want to express our sincere gratitude to all of the authors who helped write the outstanding chapters in this book. Our special appreciation goes to Author Service Manager, Dajana Pemac, for her outstanding support from the beginning to the end. The editors are also thankful to the Slovak Research and Development Agency (15-0544) and the Department of Science and Technology-Science and Engineering

diseases, epidemiology, and pain and inflammation management.

pathogenesis and therapeutic strategies of human diseases.

physiological or pathological conditions.

ments in their breakthroughs.

various diseases.


*2.2. Pecan Nuts*

Kernel halves of dry pecan nuts (*Carya illinoinensis*) from Alesto brand were purchased in a local market in Lleida (Spain). In this study, dry kernels were separated in three experimental groups comparing PEF-treated kernels against reference and control kernels. No soaking nor PEF processing was performed on reference kernels, while control kernels were placed in tap water (1:3 *w*/*w*) for 3, 20 and 35 min to assess the effect of soaking at each PEF pretreatment. Kernels pretreated by PEF were immersed in tap water and processed. After a 10 min draining, control and PEF-treated kernels (1.0 g) were separated for moisture and microstructural analysis. Before freeze-drying, kernel samples (2.0 g) were taken for determination of lipoxygenase (LOX) activity. The remaining kernels were frozen ( −16 ◦C, 24 h), freeze-dried ( −50 ◦C, 1 mbar, 72 h), and stored at −40 ◦C until oil extraction.

### *2.3. Pulsed Electric Fields Application*

The application of PEF was performed in a batch-system using a 20 × 8 cm methacrylate container with stainless-steel parallel electrodes as the treatment chamber. The batchsystem was equipped with a 0.1 μF capacitator (Physics International, USA), a pulse generator (PT-55, Pacific Atlantic Electronics Inc., El Cerrito, CA, USA) and a TG-70 gas control unit. Based on a previous study [18], dry kernels were immersed in tap water (1:3 *w*/*w*) to apply 10, 99 and 192 monopolar exponential-wave pulses at an electric field strength of 5.0 kV/cm. These electrical conditions equaled to specific energy inputs of 0.8, 7.8 and 15.0 kJ per kg of pecan nut kernels in wet basis (*W*, kJ/kg wb), respectively.


**3. Cell detection**

engineering fields. In addition to professional requirements, biomedical wafer design and manufacturing, attaches great importance to cross-cutting technology and communication skills.

The design of the chip often starts from the system or encapsulation level and starts to reverse thinking. Whether the team has sufficient cross-communication between the fields except the experts in each field and jointly solve the problems derived from each other is often the relationship between chip design and The key to success or failure Therefore, different from the training requirements of traditional engineering technology, instead it is able to cross the field of micro-electromechanical systems, microfluidics systems, biomedical technology and optoelectronic technology and other fields, it is very important in the development of biomedical wafer. When the amount of sample used to reduce, then faced with a sharp decline in signal detection problems. Increasing the signal strength or improving the sensitivity of the sensing device are two ways to solve the above problems. In terms of increasing signal intensity, there are currently artificial methods of replicating biomedical molecules to increase the weight of specimens, such as polymerization and per-chain reaction techniques; for those molecules that cannot be artificially increased, the number of markers for their markers or sensitivity, or to focus molecules in the detection area for detection. In enhancing the sensitivity of the sensing device, more sensitive new sensing technologies are also the focus of development

In this project, the hot bubble liquid bead is generated by heating the liquid by using a microheating wire and generating bubbles of great thrust in a short time to push the liquid out to form micro-droplets, as shown in **Figures 3** and **4** below. In the quantification of biomedical microbeads, the integrity and cleanliness of the beaded pellets are often quantitatively

besides reducing background noise.

204 Microfluidics and Nanofluidics

**Figure 4.** Gene microarray technology.

In cancer cell detection, gene microarray technology also has an important place. It can be used to compare the difference between normal cells and cancer cells. It can further find out which genes are more in cancer cells for further analysis. The method is to mark normal cells and cancer cells in different colors, then break them up and spread on the wafer because the DNA corresponding to the nucleotide probes on the wafer will remain on the plate. It looks like a colored plaid below.

RFLPs technology can also be applied to genetic screening. Check if embryos, fetuses or newborns have a genetic disorder. It is for quick medication. The relatives have genetic disease, but their own asymptomatic people. It can also be used by gene scanning to determine if there is a mutant gene or is likely to develop the disease. In addition, DNA Test is the most accurate way to detect a gene. It directly analyzes DNA mutations. Due to PCR (polymerase chain reaction) technology matures. DNA testing requires only a few cells to complete. It applies to one or a few mutations caused by the disease. Gene scanning and DNA detection techniques

Precisely Addressed (DNA Gene) Spray Microfluidic Chip Technology

http://dx.doi.org/10.5772/intechopen.74611

207

Sickle-shaped erythrocyte disease is a general term for a series of hereditary diseases caused by hemoglobin S (HbS). Sickle-anemia is just one of them. It arises from the point mutation in the beta-Hemoglobin gene. It causes heme β to be changed from normal glutamic acid to valine during translation. It is negatively charged by glutamic acid. Valine is electrically neutral. It makes the hemoglobin structure change. It leads to the formation of elongated needles in red blood cells. It causes red blood cells to deform, usually in severe cases. It is more minor

Sickle-shaped anemia test in two ways: First, amplified by PCR of DNA sequences, MstII restriction enzyme cutting. It is run by gel electrophoresis. Its normal sequence is cleaved by the restriction enzyme. The molecular weight is smaller and runs farther. Abnormal sequence cannot be cut by restriction enzyme, its molecular weight is larger, runs nearer. It can then check for mutations in the gene. The other is the use of artificial synthesis of two DNA test strips (one for the normal sequence "GGACTCCTC" and the other for the abnormal sequence "GGACACCTC"). It binds to normal or mutated DNA sequences amplified by PCR. Change

The human genome has about 3 billion nitrogenous bases. It contains only 5% of the DNA sequence. The rest of the sequence is mostly called junk DNA. There are more than 3 million repeats in the head. The number of these repetitive sequences varies from person to person. We call this variable number of tandem repeats (VNTRs). These repetitive sequences are ubiquitous in the genome. Its function is not yet fully understood. It is currently known to have more than ten diseases because of the excess number of repetitions in this area. For example, it is fragile X syndrome, Huntington syndrome, and the like. This area is also the main source

The genetic variation between people is only one ten thousandth. The main source of variation among individuals is the number of VNTR repeats. The number of repetitions per person varies with VNTR. When the restriction enzyme cuts the region, different individuals will produce fragments of varying lengths and numbers. We call this restriction fragment length polymorphism (RFLP). These cut-off fragments, it is different molecular weights. It has different mobility. It uses electrophoresis to separate DNA of different lengths. It goes through blotting, probe hybridization and other processes. It will show the thickness of the stripes. These stripes are called DNA

It is passed on to offspring as VNTR follows Mendel's laws of inheritance. Every pair of homologous chromosomes is in each human body. It is one from the father, one from the

fingerprinting. This was developed in 1984 by Dr. Alec Jeffreys in the United Kingdom.

are most commonly used to test for sickle-cell disease.

in color to identify if there are mutations in the gene.

of information for identifying paternity.

growth and development.

Take the example on the right, red represents cancer cells and green represents normal cells. Next, quantify the color of the statistics and calculate the ratio between them. It will find that there is significantly more red in certain areas than in green, or significantly more in green than in red. So biologists can take this to know more or less cancer cells than the average number of genes. Suppose today we find that the X gene is much more numerous than normal cells tomorrow. Biologists speculate that the X gene may be a factor in cancer. To test this hypothesis, a biologist can make a cell that has an excess of the X gene and use it to produce the protein. It can see if he has any cancer cell features. With this method we can find out the genes associated with cancer cells. This treatment of cancer cells, whether there is much to be considered for help or prevention.

Gene microarray technology is a multi-field combination of technologies as shown in **Figure 4**. It helps us better understand or crack the genetic code. The combination of technologies in many areas may be more competitive than traditional single-area research. It can solve some difficult problems in a single area.

A biological genome refers to all genetic material on the biological chromosome. Its size is often expressed as the number of base pairs. A proteome refers to the entire protein product of a gene and its performance. Many genes produce more than one type of protein after transcription and translation. There are two main reasons for this: First, a single gene undergoes alternative splicing when it is transcribed into mRNA. It leads to different combinations of exons. It thus produces different proteins. The other is that the translated protein is modified so that the protein has more different structures and functions. Proteasomes are more complicated than the genomes. In addition, a mutation of a single nucleotide (A, T, C, G) may occur at about 100–300 bases per billion of the 3 billion bases in the human genome. It thus makes the DNA sequence change, known as single nucleotide polymorphism (single nucleotide polymorphism, SNP). Therefore, the structure of DNA between individuals is very similar. The difference is very large at the micro level. It leads to significant differences among individuals. After decoding the human genome, it was found that the sequences among different individuals were very similar. It is only 0.1% different. These slight differences determine each person's height, color, size and other aspects of the difference. It also determines the different characteristics of our body. It is easy to suffer certain diseases.

We use different restriction enzymes to process the DNA. It is unique because of each individual's DNA sequence. DNA is cut into fragments of varying lengths. It is different after electrophoresis analysis. This treatment with restriction enzymes gives rise to polymorphism in length of different DNA fragments. It is called restriction fragment length polymorphisms (RFLPs). It is different from everyone's RFLPs, just like our fingerprints. Therefore, RFLPs are also called DNA fingerprinting. RFLPs technology has been widely used in the identification of criminal cases and genetic diseases.

RFLPs technology can also be applied to genetic screening. Check if embryos, fetuses or newborns have a genetic disorder. It is for quick medication. The relatives have genetic disease, but their own asymptomatic people. It can also be used by gene scanning to determine if there is a mutant gene or is likely to develop the disease. In addition, DNA Test is the most accurate way to detect a gene. It directly analyzes DNA mutations. Due to PCR (polymerase chain reaction) technology matures. DNA testing requires only a few cells to complete. It applies to one or a few mutations caused by the disease. Gene scanning and DNA detection techniques are most commonly used to test for sickle-cell disease.

which genes are more in cancer cells for further analysis. The method is to mark normal cells and cancer cells in different colors, then break them up and spread on the wafer because the DNA corresponding to the nucleotide probes on the wafer will remain on the plate. It looks

Take the example on the right, red represents cancer cells and green represents normal cells. Next, quantify the color of the statistics and calculate the ratio between them. It will find that there is significantly more red in certain areas than in green, or significantly more in green than in red. So biologists can take this to know more or less cancer cells than the average number of genes. Suppose today we find that the X gene is much more numerous than normal cells tomorrow. Biologists speculate that the X gene may be a factor in cancer. To test this hypothesis, a biologist can make a cell that has an excess of the X gene and use it to produce the protein. It can see if he has any cancer cell features. With this method we can find out the genes associated with cancer cells. This treatment of cancer cells, whether there is much to be

Gene microarray technology is a multi-field combination of technologies as shown in **Figure 4**. It helps us better understand or crack the genetic code. The combination of technologies in many areas may be more competitive than traditional single-area research. It can solve some

A biological genome refers to all genetic material on the biological chromosome. Its size is often expressed as the number of base pairs. A proteome refers to the entire protein product of a gene and its performance. Many genes produce more than one type of protein after transcription and translation. There are two main reasons for this: First, a single gene undergoes alternative splicing when it is transcribed into mRNA. It leads to different combinations of exons. It thus produces different proteins. The other is that the translated protein is modified so that the protein has more different structures and functions. Proteasomes are more complicated than the genomes. In addition, a mutation of a single nucleotide (A, T, C, G) may occur at about 100–300 bases per billion of the 3 billion bases in the human genome. It thus makes the DNA sequence change, known as single nucleotide polymorphism (single nucleotide polymorphism, SNP). Therefore, the structure of DNA between individuals is very similar. The difference is very large at the micro level. It leads to significant differences among individuals. After decoding the human genome, it was found that the sequences among different individuals were very similar. It is only 0.1% different. These slight differences determine each person's height, color, size and other aspects of the difference. It also determines the different characteristics of our body. It is

We use different restriction enzymes to process the DNA. It is unique because of each individual's DNA sequence. DNA is cut into fragments of varying lengths. It is different after electrophoresis analysis. This treatment with restriction enzymes gives rise to polymorphism in length of different DNA fragments. It is called restriction fragment length polymorphisms (RFLPs). It is different from everyone's RFLPs, just like our fingerprints. Therefore, RFLPs are also called DNA fingerprinting. RFLPs technology has been widely used in the identification

like a colored plaid below.

206 Microfluidics and Nanofluidics

considered for help or prevention.

difficult problems in a single area.

easy to suffer certain diseases.

of criminal cases and genetic diseases.

Sickle-shaped erythrocyte disease is a general term for a series of hereditary diseases caused by hemoglobin S (HbS). Sickle-anemia is just one of them. It arises from the point mutation in the beta-Hemoglobin gene. It causes heme β to be changed from normal glutamic acid to valine during translation. It is negatively charged by glutamic acid. Valine is electrically neutral. It makes the hemoglobin structure change. It leads to the formation of elongated needles in red blood cells. It causes red blood cells to deform, usually in severe cases. It is more minor growth and development.

Sickle-shaped anemia test in two ways: First, amplified by PCR of DNA sequences, MstII restriction enzyme cutting. It is run by gel electrophoresis. Its normal sequence is cleaved by the restriction enzyme. The molecular weight is smaller and runs farther. Abnormal sequence cannot be cut by restriction enzyme, its molecular weight is larger, runs nearer. It can then check for mutations in the gene. The other is the use of artificial synthesis of two DNA test strips (one for the normal sequence "GGACTCCTC" and the other for the abnormal sequence "GGACACCTC"). It binds to normal or mutated DNA sequences amplified by PCR. Change in color to identify if there are mutations in the gene.

The human genome has about 3 billion nitrogenous bases. It contains only 5% of the DNA sequence. The rest of the sequence is mostly called junk DNA. There are more than 3 million repeats in the head. The number of these repetitive sequences varies from person to person. We call this variable number of tandem repeats (VNTRs). These repetitive sequences are ubiquitous in the genome. Its function is not yet fully understood. It is currently known to have more than ten diseases because of the excess number of repetitions in this area. For example, it is fragile X syndrome, Huntington syndrome, and the like. This area is also the main source of information for identifying paternity.

The genetic variation between people is only one ten thousandth. The main source of variation among individuals is the number of VNTR repeats. The number of repetitions per person varies with VNTR. When the restriction enzyme cuts the region, different individuals will produce fragments of varying lengths and numbers. We call this restriction fragment length polymorphism (RFLP). These cut-off fragments, it is different molecular weights. It has different mobility. It uses electrophoresis to separate DNA of different lengths. It goes through blotting, probe hybridization and other processes. It will show the thickness of the stripes. These stripes are called DNA fingerprinting. This was developed in 1984 by Dr. Alec Jeffreys in the United Kingdom.

It is passed on to offspring as VNTR follows Mendel's laws of inheritance. Every pair of homologous chromosomes is in each human body. It is one from the father, one from the mother. Each person cut the fragment should be at least one of both parents the same. As long as it compares to the three-way RFLP, it can confirm the parent-child relationship.

In recent years, due to the development of polymerase chain reaction (PCR) technology. DNA identification technology uses PCR to amplify the sample DNA collected. It selects mini-satellites DNA with shorter repeat fragments in the VNTR as the target of the assay. Single-parent paternity tests can also be confirmed using maternally inherited mitochondrial DNA or paternally inherited Y chromosomes. Sperm contained in the mitochondria body is very small, most of the mitochondria within the fertilized egg from the egg. It compares the mitochondrial DNA in the offspring cells to determine if the relationship is mother-daughter, mother-daughter, or sibling. The male Y chromosome did not participate in synapsis or gene recombination during meiosis. It therefore determines the relatives of father, son, brother or paternal relative to the Y chromosome. These can be used as a paternity test to determine the relevant evidence.


*3.3.1 Male reproductive disorders*

Over the past several years, male reproductive health has been on the decline with the increase incidence of congenital malformations and poor semen quality [23, 40]. Experimental and epidemiological studies support the hypothesis that prenatal exposure to EDCs with estrogenic and/or antiandrogenic activity (e.g., diethylstilbestrol, bisphenol A, and phthalates) may disrupt the secretion and/or action of two Leydig cell hormones (testosterone and insulin-like peptide 3) regulating testicular descent, leading to cryptorchidism in newborn (**Figure 6**) [40].

**373**

**Figure 7.**

*Human Health Consequences of Endocrine-Disrupting Chemicals*

*Maternal exposure to EDCs may predispose to cryptorchidism in newborn.*

The first clinical warning with EDCs came from diethylstilbestrol, a potent estrogen mimic, given to millions of women 50–80 years ago to prevent miscarriage.

*DOI: http://dx.doi.org/10.5772/intechopen.94955*

*3.3.2 Female reproductive disorders*

**3.4 Cancers**

**Figure 6.**

and increased risk of low birth weight [25, 41].

(**Figure 7**) [2–4, 10, 12, 13, 18, 21, 22, 38, 44, 45].

*EDCs can promote the occurrence of different types of cancer.*

**Figure 6.** *Maternal exposure to EDCs may predispose to cryptorchidism in newborn.*


**1. Introduction**

Biofuels are being given serious consideration as potential sources of energy in the future. Due to recent petroleum crisis and unavailability of petroleum diesel, the need for petroleum diesel is increasing day by day hence there is a need to find out an appropriate solution. The use of renewable resources for energy production is a strategic focus of governmen‐ tal institutions to restructure the national energy economies, and many efforts are undertak‐ en to increase the share of renewable energies within the nation; biodiesel is presently the most widely accepted alternative fuel for diesel engines due to its technical, environmen‐ tal and strategic advantages. Biodiesel is known as a carbon neutral fuel because the carbon present in the exhaust is originally fixed in the atmosphere. Moreover, different vegetable oils can be used for biodiesel production like soybean oil, jatropha oil, rapeseed oil, palm seed oil, sunflower oil, corn oil, peanut oil and cottonseed oil in addition to the possibili‐ ty for use of waste cooking oil, and algal species (micro and macroalgae). Compared to fossil-based diesel fuels, biodiesel possesses many advantages such as cleaner engine emissions, biodegradable, renewable and superior lubricating property. However, until now biodiesel is not yet commercialized all over the world. And this may be due to the high cost of the raw material. Biodiesel Free on Board (FOB) costs between 0.65 and 1 U.S. dollars (USD/L). The biodiesel production cost is nearly 1.5–2 times more than petroleum-based diesel depending upon feedstock oils. It is reported that nearly 70%–95% of the total cost of biodiesel production arises from the cost of raw material; i.e., vegetable oil or animal fats. The cost of biodiesel can be reduced if we use available, non-coasted, unused biomass such as non-edible oils as frying oils, acid oils and agriculture waste oils instead of edible oils.

© 2015 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.