1.Introduction and anatomical level are similar to humans

1.Introduction
1.1 Why we select mouse model:
A mice and human both have similar tumor biology with regard to genome and functional characters, this is the reason why mouse are mostly selected for study of diverse diseases like cancers. Mouse genome is very similar to human, that’s why the basic mechanism of cancer progress and response to its cure are similar.
Different characters at cellular, molecular and anatomical level are similar to humans and have similar functioning in cancer.
1.2 RNA interference- definition:
It is a process which can prevent manifestation of gene at the phase of translation or other way is to obstruct the transcription of exact genes.
RNA intervention is basically a gene-silencing method that subdues the function of objective gene.
1.3. Advantage of this technology:
This experimental proposal offers suppleness by adjusting time and period of gene suppression.
It permits necessary functions or fatal genes to be operated in vivo by having inducible and flexible nature of gene suppression.
Quashing the gene activity is better than to knock out it.
1.4.Use of RNA interference for cure of cancer:
It is widely useful for cancer treatment. Small interfering RNA drugs for cancer has overwhelming advantages over other drugs that are used for cancer cure as well as chemotherapy. Because it acts upon the post translational stage of genic expression that’s why it does not have any harmful effect on DNA. It works by silencing many cancer sponsoring genes.
Fig 1.1 siRNA pathway

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2.Mouse model for cancer study using Small interference RNA:
Sequence specific gene silencing is carried out successfully by using small interfering RNAs. It has become an influential instrument for finding of goal and medicines. These are useful especially for cancer treatment. It can inhibit any gene of interest so we can study gene functions and can produce drugs of our demand. This technique has taken the place of knockout mouse models and the production of small molecule because many small molecule inhibitors cause toxic effects in cells.
But it also must be carefully carried out for its successful use in medicines. Enzymes like ribonucleases are used for the breakdown of siRNA which are then passed out by excretion it cannot cross cell membranes because of its high molecular weight, negative charge and water-loving capacity.
For effective delivery systems in cancer research different types of formulations are applied like liposome polymers, or micelles.
For the transfer of small particles to tumors after complete management the physical characters like size, shape, surface charge are important for concern. Long circulating nanoparticles are easily taken up by the tumor vessels because they possess irregular shapes and leaky nature. Once they are taken up the target cells via endocytosis, then they have to release in cytosol. For cancer treatment these steps are very important and should be carried out with great care from the administration site to cytosol in target cells.
Let’s take an example of orthotopic ovarian cancer for this model study, it involves in vivo transfer of siRNA
2.1 Experimentation:
2.2. Material required for experiment:
a. Commercial reagents:
0.5 M EDTA
Trizol
Glacial acetic acid
siRNA (sigma-Aldrich)
Human ovarian cancer cell lines, Hey A8, SKOVE3, A2780.
RPMI1640
Chitosan, low molecular weight
Sodium tripolyphosphate.
Hank’s balanced salt solution
Taqman miRNA assay.

2×fast syber green master mix
Verso cDNA synthesis kit
Direct-Zol RNA kit
2.3. Equipments required for experiment :
A spectrophotometer for RNA quantification
A centrifuge
A homogenizer
A thermal cycler
A real time thermal cycler
In vivo imaging system
Methodology:
How to prepare nanoparticle of siRNA/Chitosan:
Chitosan is used for many clinical applications because of its important properties like it is low toxic, biodegradable, biocompatible linear polysaccharide. SiRNA/CH nanoparticles are produced because they have ability to interact with membranes of cells. To transport siRNA into tumors the development of nanoparticle is necessary otherwise siRNA cannot cross cell membranes individually because they have high molecular weights. A process of ionic gelation is carried out for its formation using cationic chitosan and anionic siRNA and TPP.
Following steps are followed for its formation.
In 99.75 ml of water 0.25 of glacial acetic acid is dissolved to produce 0.25 % acetic acid.
Then add CH in 0.25% acetic acid to form a CH solution.
In 100ml of water 0.25 g of TPP is added to produce TPP.
Then to CH solution TPP and siRNA are added with continual mixing to form nanoparticles.
These nanoparticles are then incubated for 40 minutes at four degree Celsius temperature after incubation these particles are centrifuged for 40 min at 4 degree temperature at 14,000 rpm.
Then obtained pellet is washed to eliminate unbound chemicals then these nanoparticles are stored at 4 degree Celsius before using.
How to produce in vivo model of ovarian cancer:
First requirement is of a nude mice its age should be atleast 8-12 weeks mostly available at cancer institutes.
Second requirement is of human ovarian cancer cells like Hey A8 are cultured RPMI1640 fetal bovine serum is provided as the supplementation before they are injected into mice body.
Trypsin enzyme is used for detachment of cells then these are centrifuged at 1200 rpm for five minutes.
Now washed the cells with PBS.
Cells are then resuspended in HBBS.
30g needle is used for injecting these cells into the peritoneal cavity of mice 1×106 per 200 of µl per mouse.
Once the tumor has developed then nanoparticles are inoculated itraperitoneally in the mouse that carries tumor and then 1.25-5 microgram of dose is given per mouse.
How to isolate RNA from tissue, plasma and blood:

1.Introduction
1.1 Why we select mouse model:
A mice and human both have similar tumor biology with regard to genome and functional characters, this is the reason why mouse are mostly selected for study of diverse diseases like cancers. Mouse genome is very similar to human, that’s why the basic mechanism of cancer progress and response to its cure are similar.
Different characters at cellular, molecular and anatomical level are similar to humans and have similar functioning in cancer.
1.2 RNA interference- definition:
It is a process which can prevent manifestation of gene at the phase of translation or other way is to obstruct the transcription of exact genes.
RNA intervention is basically a gene-silencing method that subdues the function of objective gene.
1.3. Advantage of this technology:
This experimental proposal offers suppleness by adjusting time and period of gene suppression.
It permits necessary functions or fatal genes to be operated in vivo by having inducible and flexible nature of gene suppression.
Quashing the gene activity is better than to knock out it.
1.4.Use of RNA interference for cure of cancer:
It is widely useful for cancer treatment. Small interfering RNA drugs for cancer has overwhelming advantages over other drugs that are used for cancer cure as well as chemotherapy. Because it acts upon the post translational stage of genic expression that’s why it does not have any harmful effect on DNA. It works by silencing many cancer sponsoring genes.
Fig 1.1 siRNA pathway

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2.Mouse model for cancer study using Small interference RNA:
Sequence specific gene silencing is carried out successfully by using small interfering RNAs. It has become an influential instrument for finding of goal and medicines. These are useful especially for cancer treatment. It can inhibit any gene of interest so we can study gene functions and can produce drugs of our demand. This technique has taken the place of knockout mouse models and the production of small molecule because many small molecule inhibitors cause toxic effects in cells.
But it also must be carefully carried out for its successful use in medicines. Enzymes like ribonucleases are used for the breakdown of siRNA which are then passed out by excretion it cannot cross cell membranes because of its high molecular weight, negative charge and water-loving capacity.
For effective delivery systems in cancer research different types of formulations are applied like liposome polymers, or micelles.
For the transfer of small particles to tumors after complete management the physical characters like size, shape, surface charge are important for concern. Long circulating nanoparticles are easily taken up by the tumor vessels because they possess irregular shapes and leaky nature. Once they are taken up the target cells via endocytosis, then they have to release in cytosol. For cancer treatment these steps are very important and should be carried out with great care from the administration site to cytosol in target cells.
Let’s take an example of orthotopic ovarian cancer for this model study, it involves in vivo transfer of siRNA
2.1 Experimentation:
2.2. Material required for experiment:
a. Commercial reagents:
0.5 M EDTA
Trizol
Glacial acetic acid
siRNA (sigma-Aldrich)
Human ovarian cancer cell lines, Hey A8, SKOVE3, A2780.
RPMI1640
Chitosan, low molecular weight
Sodium tripolyphosphate.
Hank’s balanced salt solution
Taqman miRNA assay.

2×fast syber green master mix
Verso cDNA synthesis kit
Direct-Zol RNA kit
2.3. Equipments required for experiment :
A spectrophotometer for RNA quantification
A centrifuge
A homogenizer
A thermal cycler
A real time thermal cycler
In vivo imaging system
Methodology:
How to prepare nanoparticle of siRNA/Chitosan:
Chitosan is used for many clinical applications because of its important properties like it is low toxic, biodegradable, biocompatible linear polysaccharide. SiRNA/CH nanoparticles are produced because they have ability to interact with membranes of cells. To transport siRNA into tumors the development of nanoparticle is necessary otherwise siRNA cannot cross cell membranes individually because they have high molecular weights. A process of ionic gelation is carried out for its formation using cationic chitosan and anionic siRNA and TPP.
Following steps are followed for its formation.
In 99.75 ml of water 0.25 of glacial acetic acid is dissolved to produce 0.25 % acetic acid.
Then add CH in 0.25% acetic acid to form a CH solution.
In 100ml of water 0.25 g of TPP is added to produce TPP.
Then to CH solution TPP and siRNA are added with continual mixing to form nanoparticles.
These nanoparticles are then incubated for 40 minutes at four degree Celsius temperature after incubation these particles are centrifuged for 40 min at 4 degree temperature at 14,000 rpm.
Then obtained pellet is washed to eliminate unbound chemicals then these nanoparticles are stored at 4 degree Celsius before using.
How to produce in vivo model of ovarian cancer:
First requirement is of a nude mice its age should be atleast 8-12 weeks mostly available at cancer institutes.
Second requirement is of human ovarian cancer cells like Hey A8 are cultured RPMI1640 fetal bovine serum is provided as the supplementation before they are injected into mice body.
Trypsin enzyme is used for detachment of cells then these are centrifuged at 1200 rpm for five minutes.
Now washed the cells with PBS.
Cells are then resuspended in HBBS.
30g needle is used for injecting these cells into the peritoneal cavity of mice 1×106 per 200 of µl per mouse.
Once the tumor has developed then nanoparticles are inoculated itraperitoneally in the mouse that carries tumor and then 1.25-5 microgram of dose is given per mouse.
How to isolate RNA from tissue, plasma and blood:

1.1.1) Piaget proposed the theory of learning through four stages of cognitive development which reflected on the increasing sophistication of childrens thought ( McLeod , S.A ,2015) .He also suggested that the theory involved the 3 processes such as assimilation, accommodation and equilibration ( Luneta, 2013: 30) . Bruner on the other hand suggests that the theory of learning does not only include the concepts and problem- solving procedures invented previously by the culture, but also the ability to “invent” these things for oneself. This theory involves three modes of representations namely enactive, iconic and symbolic ( McLeod , S.A , 2008) .
1.1.2) Bruner outcome of cognitive development is thinking. The intelligent mind creates from experience “generic coding systems that permit one to go beyond the data to new and possibly fruitful predictions ” ( Bruner , 1957: 234). Whereas Jean Piaget’s theory of cognitive development focusses on how learners interact with their environment to develop complex reasoning and knowledge.
QUESTION 1.2
Nature is ones genes such as the different types of characteristics a child develops / inherits from their parents. Nature is therefore responsible for the biological influences in a child’s development such as appearance, talents, abilities and also certain illness. It is also responsible for the growth of a person from the fetus level until development into a normal adult ( Groark , McCarthy & Kirk , 2014) .Nurture is what a child has been taught or what they learnt from people around them such as manners like please and thank you either by parents or teachers. Nurture influences children’s development through many ways: physically through nutrition, activity, and stress; intellectually through informal experiences and formal instruction; and socially through adult role models and peer relationships. With good environmental support, children thrive to be the best they can be. Unfortunately, the conditions of nurture are not always the best. For example, children who grow up in a cruel and violent family must look outside the family for love and care ( McDevitt, Ormrud,2007: 6-8). Guidance from parents and teachers plays a very important role in childrens lives. Teachers promote self determination which allows students to have confidence in themselves and strive towards their goals and succeed in their lives. Teachers who are able to inspire, support and provide appropriate care for their students certainly cause a great impact in their lives. A teacher not only has the ability to provide knowledge, but also guide children how to ask the right questions and develop skills needed to succeed in life (2016). Whereas a parent’s role is not only to be kept to the academic and educational achievements of a child, but also in the development of inner personality and character of a child. Their influence will be very high with regards to the entire development of children. Starting from relations to education, everything starts in the home itself and parents play an important role in developing many qualities of a child. A parent’s abilities, aspirations, habits, behaviour etc. are some of the factors which contribute to success or failure of child (2013). Through their culture children learn and realize which behaviours and temperaments are allowed, discouraged and forbidden. Traditions, religion, food and even the manner in which a particular culture dresses influences child development. According to various researches it suggests that cultural socialization within families helps to develop high self-esteem, racial and ethnic pride (Groark, McCarthy, ; Kirk, 2014, ch.2.3). It is therefore important for educators and parents to educate their children to understand how culture shapes child development in order to improve the lives of children in a way that respects their cultural backgrounds (2012).

1.1.1) Piaget proposed the theory of learning through four stages of cognitive development which reflected on the increasing sophistication of childrens thought ( McLeod , S.A ,2015) .He also suggested that the theory involved the 3 processes such as assimilation, accommodation and equilibration ( Luneta, 2013: 30) . Bruner on the other hand suggests that the theory of learning does not only include the concepts and problem- solving procedures invented previously by the culture, but also the ability to “invent” these things for oneself. This theory involves three modes of representations namely enactive, iconic and symbolic ( McLeod , S.A , 2008) .
1.1.2) Bruner outcome of cognitive development is thinking. The intelligent mind creates from experience “generic coding systems that permit one to go beyond the data to new and possibly fruitful predictions ” ( Bruner , 1957: 234). Whereas Jean Piaget’s theory of cognitive development focusses on how learners interact with their environment to develop complex reasoning and knowledge.
QUESTION 1.2
Nature is ones genes such as the different types of characteristics a child develops / inherits from their parents. Nature is therefore responsible for the biological influences in a child’s development such as appearance, talents, abilities and also certain illness. It is also responsible for the growth of a person from the fetus level until development into a normal adult ( Groark , McCarthy & Kirk , 2014) .Nurture is what a child has been taught or what they learnt from people around them such as manners like please and thank you either by parents or teachers. Nurture influences children’s development through many ways: physically through nutrition, activity, and stress; intellectually through informal experiences and formal instruction; and socially through adult role models and peer relationships. With good environmental support, children thrive to be the best they can be. Unfortunately, the conditions of nurture are not always the best. For example, children who grow up in a cruel and violent family must look outside the family for love and care ( McDevitt, Ormrud,2007: 6-8). Guidance from parents and teachers plays a very important role in childrens lives. Teachers promote self determination which allows students to have confidence in themselves and strive towards their goals and succeed in their lives. Teachers who are able to inspire, support and provide appropriate care for their students certainly cause a great impact in their lives. A teacher not only has the ability to provide knowledge, but also guide children how to ask the right questions and develop skills needed to succeed in life (2016). Whereas a parent’s role is not only to be kept to the academic and educational achievements of a child, but also in the development of inner personality and character of a child. Their influence will be very high with regards to the entire development of children. Starting from relations to education, everything starts in the home itself and parents play an important role in developing many qualities of a child. A parent’s abilities, aspirations, habits, behaviour etc. are some of the factors which contribute to success or failure of child (2013). Through their culture children learn and realize which behaviours and temperaments are allowed, discouraged and forbidden. Traditions, religion, food and even the manner in which a particular culture dresses influences child development. According to various researches it suggests that cultural socialization within families helps to develop high self-esteem, racial and ethnic pride (Groark, McCarthy, ; Kirk, 2014, ch.2.3). It is therefore important for educators and parents to educate their children to understand how culture shapes child development in order to improve the lives of children in a way that respects their cultural backgrounds (2012).

1.0 Introduction.
According to Eurostat (2017) explained that water is essential for life and can be referred as surface water and ground water and also there is renewal water resource which calculate the internal flow which comprises precipitation minus actual evaporation. Water can obtained through the combination of hydrogen and oxygen which results to water. Water has been good service both human being and other living things, water is on demand for life on earth because every living organism require water in order to survive. The source of fresh water is associated with the presence of forest and mountainous ecosystem which contribute largely on the presence of fresh water (Reid et al., 2005). The world population has increased and there is high demand for water resource use though there is increase of water scarcity compared to the ratio of the population consumption on water resource. Therefore the aim of this study will be to speculate factors for water scarcity and demand with the ratio of population, challenges facing water utility with the current population increase in the city areas also the study will find out the measures to be taken sustainable and accessible of water resource for economic and social development.
1.2 Background of the research problem
The urban population of the world is forecasted to reach 6.3 billion people from 3.4 billion in 2009, in which it present population growth and net migration. This increased population in the urban areas stress the demand of water with high consumption, also the urban population lack access to improved water supply and sanitation up to 20% since the millennium Development Goals has been established. The water demand in the world has been dramatically increased due to the increase number of people with high demand to water resource also due the development of economic activities such as agriculture , production of energy , industrial uses and human consumption is very high (UN Water 2008). The world population for supply of water is approximately to reach four billion people and sometimes up to two thirds of the global population. The current trend incur the global climate change has led to the threat on sustainable water suppl. the global fresh water has been expanding up to a mean rate of about 20% between 1960 and 2000 , in which the water use has doubled in one decade. The use of water by human has been unattainable. From 5% to 25% of global freshwater use exceeds long term accessibility and about one billion people live in areas with no good access to freshwater to sustain their life. Therefore also it has been explained that water use is predicted to grow by approximately 10% between 2000 and 2010 compare with rate of 20% over the past forty years (Reid, et al2005).

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