Zebrafish in Toxicology Research: Advantages and Limitations

Zebrafish in Toxicology search Advantages and LimitationsThe ab ingestion of psychedelic substances is a serious macrocosm health problem and understands the brain disorder bring on by mind-expanding substances is an all- meaning(a)(a) challenge in biomedical question that requires extensive clinical and preclinical investigating (Neelkantan et al. 2013 Stewart et al. 2011). Recreate the complex network of neuro chemical substance interactions in organisms in vitro is non possible, especially for psychotropic medicates (Riehl et al. 2010). In this way, experimental brute feignings is a burning(prenominal) tool around for investigating of the toxicology and chemistry of the drugs of ab utilize.Animal mystifys in toxicologic and pharmacologic research employ mammals present high cost, consumes large amounts of chemical compounds and ar laborious to perform the tests and primary(prenominal)tenance of animals. In this context, animal places such as the zebrafish, w hich outweigh these disadvantages, be an interesting and promising alter inhering in research.Since the 1970s, the zebrafish (Danio rerio) is used in research in neuroscience and ontogenyal biology (Serra et al. 1999). In the 1990s, zebrafish were used for the first vertebrate large scale mutagenesis screen. In stick out few age, the use of zebrafish (Danio rerio) in scientific research has affixd speedily in other science fields, including Pharmacology and Toxicology (Chakhaborty et al. 2009).The zebrafish represents an interasting model for integration of research of genetic, neural and behavioral aspects (Miklsi and Andrew, 2006). The coupling between behavioral assays and analytical and molecular techniques permits the elucidation of mechanisms of toxicity, the test of pertly drugs for therapeutic treatments and the study of refreshing drugs (Tierney et al. 2011). The aim of this paper is to outline recent developments and futures spots in drugs of abuse research with z ebrafish, in Experimental and Analytical Toxicology.Zebrafish as a impudent animal model in Toxicology advantages and limitationsThe zebrafish is a humbled tropical fish native of northern India and adjacent countries. Many factors and animal characteristics settle zebrafish an attractive and in effect(p) model to dismember the mechanisms of action and set up of drugs in general (Chakhaborty et al. 2009). Zebrafish and serviceman shargon about 75% of their genome and scram physiological identicality (Chakhaborty et al. 2009, Zhu et al. 2014). Genome, transcriptome and proteome of zebrafish use up been panopticly studied and described (Maximino et al. 2010). The similarity level between zebrafish and world is withal observed in the nervous system (Mathur and Guo, 2010).Zebrafish are small (at adulthood, 2.5 4.0 cm) and maintenance costs of this fish in laboratory are substantially low, permitting logistical and economical advantages oer rodent models (Key and Devine, 2 003). The fertility rate and the fleck of embryos generated are higher those in mammalian models (Chakhaborty et al. 2009). In general, an adult pistil tardily can produce 200-300 eggs and reproduce 2 or 3 times per week (Blaser and Gerlai, 2006 Gerlai 2003 Patton and Zon, 2001 Zon and Peterson, 2005).The zebrafish cycle of manner and development period is well-characterized and brusque (3 to 4 months) (Cadet, 2009 Hill et al. 2005). The body is formed in the first 24 hours post fertilization (hpf) and the internal variety meat are fully develop at 96 hpf (Chakhaborty et al. 2009 De Esch et al. 2012b Ninkovic and Bally-Cuif, 2006b Parng et al. 2002 Patton and Zon, 2001). The development outside of the uterus and the optical hydrofoil of eggs and tissues during embryogenesis allows the opticization of tissues and organs in vivo, do possible the visual analyses in real time of early developmental kneades, organ syllable structure and dysfunctions caused by drugs of abuse an d quantification of cell proliferation and cell death in special(prenominal) tissues (Chakhaborty et al. 2009 Friedrich et al. 2010 Hill et al. 2002 Ingham 2009 Miklsi and Andrew, 2006 Mathur and Guo, 2010 Ninkovic et al. 2006b Parng et al. 2002 Patton and Zon, 2001 Peterson and MacRae, 2012 Xu et al. 2011). In addition, blastomeres of zebrafish are large and stable for biophysical and electrophysiological assays (Zhu et al. 2014).The zebrafish has flex a widely utilized model organism in pharmacological and toxicologic research, particularly collectible to evidence that they may share with gentleman and other mammals some key receptors targeted by drugs of abuse (Miller et al. 2014). In addition, zebrafish are highly cordial animals which enables them to display robust behavioral responses, such as shoaling, aggression and social preference, and emerge as a sensitive selection model to investigation of drugs of abuse-evoked states (Cachat et al. 2013 Pham et al. 2012). Anothe r advantage is that the size of zebrafish provides a quick absorption of drugs via the gills and leads to bioaccumulation in Central Nervous System (CNS) and other tissues (Echevarria et al. 2008).Zebrafish is manageable to molecular and genetic analysis (Bailey et al. 2013 Chakhaborty et al. 2009 Miklsi and Andrew, 2006 Parng et al. 2002). Since genetic mutations can intercede in brain function, the use of molecular and biochemical techniques in zebrafish allows the recognition of molecular substrates for drugs in brain and the analysis of function and regulation of the genes, the business of transgenic strains and the induction of detail mutations, inducing all overexpressing of the genes or decreasing genes expression (Cadet 2009 gold-worker 2004 Key and Devine, 2003). Use of zebrafish mutant strains has allowed the discernment of mechanisms and pathways and neural expression of proper(postnominal) genes. ascribable to the counterchange magnitude progress in this area , studies of mutant zebrafish halt investigated specific behaviors, diseases, deformities and operable processes (Spitsbergen and Kent, 2003). The performance of point mutations in zebrafish, generating mutant lines, enables the molecular investigation of the mechanism of action of drugs of abuse, determining specific receptors and target molecules.Although the model advantages are numerous, some limitations should be considered in the use of zebrafish in toxicological, pharmacological and neuroscience research. The rule or defective functioning of the human brain is complex to be fully modeled in zebrafish brain. Some areas do not fuddle the same degree of development of the mammalian brain, ma office it fractious to chromosome mapping for mammalian counterparts (Kalueff et al. 2014 Rinkwitz et al. 2011 Stewart et al. 2014). Moreover, the social behavior is not clearly defined in fish larvae (Kalueff et al. 2014 Stewart et al. 2014).Zebrafish has a genome 30-40% tetraploid du e to an genome duplication with some of their genes having two copies kinda of one as in mammals and one of these two genes are not represented in the human genome (Alestrm et al. 2006 Klee et al. 2011 Stewart et al. 2014).One disadvantage associated with the zebrafish model is the fact that some drugs are not soluble in pissing, making it difficult to institution by immersion. This potential problem can be solved by using organic solvents or other routes of administration (Stewart et al. 2014). In other hand, exposition to drug by immersion reduces the nisus-induced by injection (Stewart et al. 2011b). Finally, although the barrier between brain and fund presents development similar to humans, the distribution of some drugs in organism may be contrastive in diverse species (Stewart et al. 2014).Although the pattern has some limitations, the advantages over conventional models mammals make zebrafish a very interesting alternative in research. In this sense, the use of zebra fish in research involving drugs of abuse, in order to characterize drugs of abuse set up and to quantify these compounds in a biological fluid from the animal is increasing and represents a great target for research.Zebrafish research to study the drugs of abuseThe endure of toxicology research in laboratory animals has make up a well-established and essential perpetrate and chemical and toxicological information on chemical compounds and drugs are obtained from the results of these studies (Gad 2007 Olson et al. 2000). The use of animals in research has several advantages low cost, easily use and functional homology with humans (Gerlai et al. 2010).The animal experiments to predict the action, metabolism and effects of drugs of abuse in humans are extremely important in Toxicology.Introduced as a model for neural development by George Streisinger in the 1960s, zebrafish has become a promising aquatic model for study of drugs. The zebrafish is an efficient alternative model of drug delivery via the gills, by direct application of drugs in water and posterior submersion of the animal. Between 12 and 14 days by and by fertilization (dpf), molecular oxygen is in general absorbed by the skin, counseling that this is the main route of uptake for small molecules, and after 72 h the embryos go about to swallow indicating the availability of the oral route for absorption of chemical compounds (Goldsmith 2004). exogenous compounds, such as ethanol, have been shown to rapidly enter in systemic circulation of the fish, demonstrating the high sensitivity of the zebrafish to various psychotropic agents (Dlugos and Rabin, 2003 Echevarria et al 2006 Kyzar et al. 2012). In the larval and embryonic stage, the performance of rapid and high-throughput analysis of multiple behaviors and screening of chemical compounds is possible and competent (De Esch et al. 2012b Guo 2009 Richendrfer et al. 2012).The first response to the action of a psychoactive substance is the beh avior. In this sense, the zebrafish has been widely used in the development and institution of behavioral assays to different drugs of abuse, due to its sensitivity to drugs and their robust responses. Locomotor activity, learning, sleep, aggression, social and antipredator behavior are some of the observed and well-characterized behavioral parameters in zebrafish (Bailey et al. 2013Cachat et al. 2013 Guo 2004 Pham et al. 2012 Spitsbergen and Kent, 2003). ductless gland responses to stress in zebrafish are also an important tool for toxicological effects induced by drugs of abuse. In zebrafish, the hypothalamus-pituitary-interrenal axis (HPI) is homologous to the hypothalamic-pituitary-adrenal axis (HPA), found in mammals and humans, and cortisol is involved in physiological responses to stress situations (Stewart et al. 2010).Another interesting pattern of physiological change for toxicological investigation of drug in zebrafish is the color of the skin. As mammals, fish have cel ls containing pigments in the collagen layer of the dermis and these cells have hormonal and nervous regulation (Nguyen et al. 2013). In this sense, effects on skin color may contribute in part of the elucidation of the mechanism of action of many drugs of abuse.The toxicity induced by abuse of drugs in the digestive and cardiovascular systems of zebrafish can also be determined. These systems are developed and become fully functional in the first days after fertilization (Patton and Zon, 2001). The characterization and monitoring of blood vessels in embryos is easily achievable (Rubinstein 2003). The heart rate is an indicator of cardiac toxicity widely used. Morphological examination of the heart and digestive organs can be performed to identify organ-specific toxicity of drugs of abuse (Rubinstein 2006). For example, the effects of environmental toxin tetrachlorodibenzo-para-dioxin (TCDD) on heart rate (Henry et al. 1997), the functional lipid metabolism (Farber et al. 2001), the effects of fungicide riphenyltin acetate (APTT) in hepatocytes (Strmac and Braunbeck, 1999) were investigated using histological and immunochemical analysis.The development of analytical techniques to determine compounds of interest in biological fluids zebrafish also plays an important role in the elucidation of mechanisms of action and toxicity of drugs of abuse. Moreover, it is extremely important to develop methods of extraction and concentration of analytes and biomarkers that make possible dosages required to understanding toxicological aspects of these drugs of abuse. The main analytical methods developed to study the drugs of abuse in zebrafish are listed in Table 1.The new psychoactive substances (NPS) that have emerged in recent years have action and effect mechanisms partially or essentially unfathomed. harmonise to UNODC, the number of NPS significantly increase between 2009 and 2013 (UNODC 2014). In many poisoning deaths caused by these unknown substances, the lack of information makes it difficult to identify the cause of death. Therefore, the use of zebrafish in research represents an important tool to scan and evaluate the chemical and toxicological properties of both known and unknown drugs of abuse.Marketing Design and Innovation WrigleyMarketing Design and Innovation WrigleyWrigley is a well-known global shuffle dealing in wide run for of confectionery intersections such as chewing gums, chewy and hard candies, mints, and lollipops. It is operate in nearly 50 countries and distributing its ingatherings in much than 180 countries. The gild is based in Chicago, Illinois and was founded in 1891. Some of its check offs i.e. Juicy Fruit, Wrigleys Spearmint and Altoids have heritage of more(prenominal) than hundred years. Wrigley is famous for its innovative crossings that are introduced into its return line from time to time. Wrigley uses beat out ingredients and packaging materials for its wares and the core principles on which it operates implicate quality, exemptdom, efficiency, responsibility and mutuality (Wrigley.com, 2017).Late in 2014, Wrigley merged its Orbit and superfluous brand but it launched special with new im prove taste in 2015. As per the new foodstuff trends for health and taste, additional is a new sugar-free gum forthcoming in single packs, multipacks and bottles introduced in more than fiver different tastes. The harvest-home is launched for Australian and New Zealand mart. One of the better(p) attributes of Wrigleys product is innovation and the concern for consumers health. Wrigley continuously update the flavors of its limited brand to satisfy the palate of its valued customers.UK confectionery market have been observed to have a high volume consumption and it stands as the sixth largest in terms of confectionery consumption. The confectionery market is driven by pleasure, convenience, drug replacement, snacking and innovation in products (Globaldata, 2016). Consumers a re becoming health conscious due to the increasing issues like diabetes and obesity. Dentists are also increasing the awareness among battalion about oral health and the tooth related diseases such as brass instrument and saliva generation etc. dentists are making the consumers aware of the benefits of chewing. Due to the increased health consciousness, the great unwashed are likely to take more products for treatment and prevention of such issues.Wrigley has launched surplus and Extra nimble in standstill with The Australian Dental link (ADA), New Zealand Dental Association (NZDA) and the FDI World Dental Federation. According to Country Report Wrigleys Extra attach the take onth in 2016 because of its sugar-free and oral care features (Country Report, 2016). Now, Extra is involved in cause-related market and is making associations with the dental consonant professional person to educate the families about the consequence of oral health. This brand contributed about one tierce of the helping of total value shares of the Wrigley Company and is expected to continue to gain highest share because of its uncountable benefits (Country Report, 2016).Maslow pecking order of Needs theory According to the Maslow hierarchy of inevitably theory, Wrigleys product Extra attempts to satiate the physiological of necessity of an singular. Although its not radical necessity for life but oral health and refreshment are necessary for an single and Extra is capable of meeting these needs. It also satisfies the social needs of an individual because it boosts the confidence and reduces pressure and anxiety agree to the research conducted by psychologists (Smith, 2012). lively breath and sweet smell is necessary if an individual is attending a social gathering or event which is satiated by Extra i.e. to meet the love and belonging needs. People with high level of income commonly buy Wrigleys Extra which also satisfy esteem needs because it represents the post ure quo of the individuals consuming this brand. The self-actualization needs of an individual can be satiated by Extra because it gives a new touch to the lifestyle of an individual i.e. a refreshing breath and increased confidence. scattering of Innovation ModelDiffusion is defined as the process by which an innovation product is passed on to the members of a social system with the help of different channels over a specified time period. There are five types of passel involved in the diffusion of innovation process i.e. innovators who are venturesome and are ready to the test the product as soon as possible early adopters adopt the liking and are aware of the benefits associated with change early majority adopts the idea before an average person does late majority adopts the idea once a greater majority has time-tested it, and laggards are cynical to change and do not adopt the idea usually (Smith and Song, 2004).Due to increased awareness among consumers about oral health, th e percentage of lot using Extra brand is increasing. Wrigleys Extra has been rated 8.9 out of 10 by the consumers which means that they are early adopters who have adopted the product within a time span of two to three years (infoscout.co, 2017). The other reason for product adaptability is the range of tastes on tap(predicate) i.e. the consumers consume different tastes of the products to please their palates. Consumers prefer products which are healthy and Extra is healthy in terms of oral care and its sugar-free as well. Extra is clinically proved by the professional dental associations and it also stimulates the saliva by 10 times.It is very important to identify the needs of consumers and become aware of the market trends before launching a new product. Because the consumers are becoming increasingly health conscious, Wrigley has identified recent trends in the market and launched Extra into the market.Kano modelCustomers needs change over the period of time due to the ever-c hanging market trends and in the recent business environment, customer is considered as the king of a business. According to the Kano model, a product must be developed considering the basic needs of customers, performance needs and excitement needs i.e. basic needs are hygiene needs turn excitement needs are delighting needs and these delight needs later one become basic needs and then customers need something new (Wang and Ji, 2010).Wrigleys Extra is a product that can satisfy the fun and pleasure needs along with the health needs of an individual. The hygiene needs of Wrigleys Extra are the onward motion of oral health, prevention of teeth caries, and organisation reduction etc. while the fun and pleasure attributes of Extra satisfy the delight needs. With the innovation of brand these delight needs will become the basic needs of an individual. The merchantman line is that the product should have the attractive and must-be attributes that can fulfill the needs of customers fo r which they are using the product. These attributes of Extra are health and bacteria prevention ingredients.Product numberDuring the product development, the anatomy of product is important to analyze in terms of the benefit it will offer (Blythe, 2017). The product anatomy is conducted to match the consumer trends with the product offered by a particular brand. The core product of Wrigleys Extra is the oral health and its sugar free i.e. cleans food debris, neutralizes plaque acids, strengthens teeth, reduces dental caries, prevents tooth decay, and stains. The generic product of Wrigleys Extra is that its available in wide range of flavors i.e. strawberry, peppermint, spearmint, white lemon lime, bubble mint, water melon, berry and chromatic cream etc. The expected product is that it stimulates saliva and refreshes individuals breath. And the augmented product of Wrigleys Extra is that it is available in different types of packaging i.e. 14 pieces and 10 pieces single pack, 28 and 42 pieces multipack, and 46 pieces bottle (Desfosse, 2017).This product has pleased the palate of numerous customers which is reflected in the increased gross sales radiation pattern of Wrigley. A report showed that one trey of Wrigleys sales of 2016 were contributed by Extra (Country Report, 2016). It shows that people are continuously using the Wrigleys Extra for their oral health.The development of Wrigleys Extra has shaped the overall gum market and has proved to be a market leader in 2016 by increasing the overall sales of Wrigley. The prime cause for increased value of Extra is its association with the professional dental associations and being a sugar-free chewing gum. The brand awareness is created by educating the families and general public through the use of creative and educative advertisements (Schultz, 2015).According to statistics of 2015, the sugarless and breath fresheners has seen considerable increase in dollar and social unit terms. The sales of Wrigleys Extra increased to 9.3% from 8.6% a year ago. This is because consumers are adopting the more socially acceptable mint and refreshment gums for refreshing themselves. The sales of Wrigley Extra are expected to grow in the near future because the dental care specialists are positioning the oral care and sugar free gums as more important for the consumers to chew after and between the meals (CSP mundane News, 2015). It will be easy for Wrigley to retain its customers for Extra because of the cause-related marketing and its association with the professional dental associations. Due to the increased consciousness and awareness among the people about obesity, they are reducing more and more on the snacks and this is the best alternative that can be replaced for snacks. This trend of marketing is likely to increase the sales figure for Extra chewing gum. Another important marketing milestone of Extra is that its associated with the Action for Children charity donations from sales of Ext ra packs. equilibrize scorecard credit line scorecard is the metric used for the purpose of reorient different strategies with the vision and mission of an organization. It undertakes the newly formed strategy of a particular business and transforms it into marching order on daily basis. A newly developed product can be seen from four perspectives according to the balanced scorecard. These four perspectives are learning and growth perspective, business process perspective, customer perspective and financial perspective (Violet and Josiah, 2011).Learning and growth perspective According to the recent statistics for Extra gum, it can be evaluated that the consumers percentage using this product is increased a certain extent during 2016 and is expected to grow further (Country Report, 2016). Although Extra gum was once merged with the Orbit brand of Wrigley but later on it was relaunched with improved taste and packaging that attracted a whole new lot of consumers and increased the ma rket share by 71%. Customer are satisfied with Extra and Extra Active brand of Wrigley due to its string oral care positioning and sugar-free attribute.Business process perspective Extra gum was launched with new taste and packaging and it was brought back to the market due to its increased demand from the customers. Management has launched the product after conducting complete market research and clinical importance of the product. It is make with best possible ingredients i.e. real sugar and no artificial modify are added into it. The unit sales of Extra gum are increased from 8.6% to 9.3% that shows the efficient procurement and distribution capabilities of Wrigleys Extra (CSP Daily News, 2015).Customer perspective Extra is available in different tastes, different packets size and is suitable for customers of any age. The product is best for chewing before and after the meals and is mainly targeting the oral care of children and aged people. It is the best replacement for the cu stomers who want to chuck up the sponge smoking and it can also be used after smoking. Customers are satisfied about this product which is evident from the increased sales figure and the reviews of customers for this product (Wrigley.com, 2017).Financial perspective Extra gum has contributed one third of the total sales of Wrigleys overall sales figure. The Country Report (2016) quotes that The friendship is expected to edge up by around a third of a percentage point overall to reach close to 89% value share. The companys share gain in 2016 will be solo due to its Extra brand, which alone will gain over a percentage point in the year and dominate sales with over 71%. It means that the brand is also performing well in terms of finance.The overall position and performance of Extra gum suggest that due to the oral care positioning and cause related toil and marketing of the product, Extra will be put as an important element for the oral care regimes. The competitive advantage of th e product is that it is available in different sizes, flavors, packets, is nutritional and perfect for all age users and is available in more than 180 countries.Extra gum is one of the best products designed by Wrigley for oral care and is sugar-free which was once kicked out of the market due to the other brands offering same product. Based on the heritage of more than hundred years and the best quality, Extra reclaimed its leading and innovative place in the market after its launch with new tastes and packaging. The innovation and leadership attributes of Wrigley are renowned in the market. The company should reach out its best and keep an eye on the market to continuously cost increase the brand in terms of packaging and tastes so that it may not lose the market share.ReferencesBlythe, J. (2017). Product Anatomy. Online SAGE Books. Available at http//dx.doi.org/10.4135/9781446221617.n17 Accessed 18 Mar. 2017.CSP Daily News. (2017). 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