anxious and nervous

Have you ever felt so anxious and nervous that you would go to any extreme to relieve that feeling? Well, downers are drugs that many people use to relax and bring themselves down from feeling anxious or nervous. These drugs not only make you feel less anxious or nervous but they depress the central nervous system and produce a relaxing to sleep-inducing effect depending on the drug dose that is taken. Downers also slow down and depress the level of arousal in the brain by slowing down the nerve cells in the central nervous system, dull thinking abilities and reduce inhibitions. Downer drugs are not all that bad. Some of these drugs are used for medical reasons, actually prescribed by doctors. Some of the sicknesses that downers are used to treat include insomnia, to help manage anxiety, to reduce convulsiveness, and for neuromuscular disorders. They are most often prescribed today as tranquilizers and sleeping pills. Some examples of tranquilizers are Valium, Librium, and Xanax. Some examples of sleeping pills include Dalmane, Halcion, and Resoril. It is said that over the last decade an estimated six hundred million prescriptions were processed for minor tranquilizers in the United States pharmacies alone. Downers, which were once thought to be the perfect sedative for the terminally nervous, insomniacs, epileptics, mental patients and also used for anesthesia before surgery is now found to be very addictive. Downers as a whole have many risks and side effects to their users. Being downers are a depressant, they reduce cognitive and motor skills and decrease judgment and increase reaction time (meaning the time it takes to react increases.) Subtle effects can continue for hours or days after the use of downers because of the length of time it takes to break down some depressants in the body. Because of a very high abuse to downers, manufactures now warn users against using sleeping pills for more than a week ...

Tunku Abdul Rahman Essay Example

Tunku Abdul Rahman Essay Example Tunku Abdul Rahman Essay Tunku Abdul Rahman Essay Essay Topic: Maus Sir Tunku Abdul Rahman Putra Al-Haj ibni Almarhum Sultan Abdul Hamid Halim Shah was the Chief Minister of the Federation of Malaya from 1955 and became the countrys first Prime Minister after its Independence in 1957. Born in a royal family in Kedah, Abdul Rahman was the son of 24th Sultan of Kedah and went to become the founding father of the independent Malaysia. Before attaining independence of the country, Abdul Rahman was also a member of the United Malays National Organization, an organization struggling against Britains Malayan Union. Also known as Bapa Kemerdekaan, he became the first Prime Minister of the country after Sarawak, Sabah and Singapore merged to form Malaysia in 1963. He played a key role when the Organization of Islamic Conference was established in 1969 and served as its first Secretary-General. Childhood Early Life Abdul Rahman was born on 8 February 1903 in Istana Pelamin, Alor Star in Kedah to the 24th Sultan of Kedah Sultan Abdul Hamid Halim Shah and his sixth wife Cik Menjalara. He was the fourteenth son and twentieth child of his father. As a child Abdul Rahman was raised in Isatan, the Palace, which was built by a Chinese contractor and had a royal childhood with a number of servants around him. Prince Abdul was sent to a Malay Primary School Jalan Baharu in 1909 and was later moved to the Government English School in Alor Star, which eventually became known as the Sultan Abdul Hamid College. In 1911, little prince Abdul was sent to Debsirin School in Bangkok to study with his three brothers. He returned to Malaya in 1915 and resumed his studies at Penang Free School. After three years in 1918, he enrolled at St. Catharine’s College in Cambridge University with the Kedah State Scholarship and graduated with a bachelor degree in Arts in 1925. At that time, he drew national attention and was praised as the first student to receive a scholarship from Kedah State for studying in the United Kingdom. Early Career After finishing his graduation, Abdul Rahman served in the Kedah public service and was made the District Officer of Kulim and Sungai Petani. At that time, the colonial Malaya was totally dominated by the British officers, with the only exception of Abdul Rahman who was a Malaya and had concern for his fellow people of Malaya. The two could never come to terms and he often had to pay for it by loosing promotion to higher positions. However, the British Administration did not dare do anything against him, as he was a son of the Sultan and a price of Malaya. Few years later, Abdul went to England and stayed there briefly before finally returning to Malaya following the eruption of the World War II. He continued his law studies at Inner Temple in 1947 and after admitted to bar in 1949, he returned to Malaya where he was appointed at the Legal Officer’s office in Alor Star in the same year. Further, he was made a Deputy Public Prosecutor in Kaula Lampur and president of the session’s court. Abdul Rahman was also a member of the United Malays National Organization, an organization struggling against Britain’s Malayan Union. In 1951, a conflict surfaced within the UMNO forcing its president Datuk onn Jaafar to resign and the person replacing him was Abdul Rahman, who eventually seized the position for the next twenty years. Independence of Malaya Abdul Rahman began his campaign for Malaya’s independence in 1954. His initial efforts failed to achieve anything as the British Administration was averse to grant independence unless it was assured of the racial harmony and equality in a new independent Malaya. As President of the UMNO, Abdul Rahman consolidated a political alliance with the Malaya Chinese Association to form the Alliance Party and later the Indian community Malayan Indian Association joined them in 1955. The coalition gained huge popularity among people though the UMNO members were initially reluctant to open the party to Chinese and Indian community. Abdul Rahman became the first Chief Minister of Malaya with the Alliance Party winning the first federal general election held in the same year. In 1955, he traveled to Japan, where he negotiated the Malaya Independence and 31 August 1957 was finally decided for its independence. Premiership of Malaysia Malaya became Malaysia with the emergence of Singapore, Sabah, Sarawak and Brunei in 1963. In 1961, Abdul Rahman called upon these states to form an amalgamation and was subsequently elected the first Prime Minister of Malaysia on 16 September 1963. However Singapore’s addition in the federal proved disastrous which increased the Chinese influx in the country. Abdul Rahman fearing, that Lee Kuan Yew’s Party could influence the voters in Malaya, started demanding Singapore’s exclusion from Malaysia. After endless clashes between Abdul Rahman and Lee Kuan, Singapore seceded and declared its Independence on 9 August 1965. Abdul Rahman’s regime came to downfall in 1969, when the Alliance Party lost most of its support in the general election that year. Abdul Rahman lost its support from the people within UMNO who were highly critical of his headship and eventually, an emergency committee captured the country from Abdul Rahman and declared a state of emergency leaving him with no power. On 22 November 1970 Abdul Rahman resigned from the position of Prime Minister and subsequently from UMNO in 1971. Other Activities In 1960, when he was still the Chief Minister, Abdul Rahman declared Islam the official religion of Malaysia and established the Islamic Welfare Organization (PERKIM) as a guiding body for Muslim converts. He became the president of PERKIM and served until a year before his death. As President of PERKIM, he organized the first International Quran Recital Competition in 1961. Abdul Rahman played a key role when the Organization of Islamic Conference was established in 1969 and served as its first Secretary-General. He was a co founder of the Islamic Development Bank and President of the Regional Islamic Da’wah Council of South East Asia and the Pacific (RISEAP), serving from 1982 till 1988. However he declared Islam the official religion of Malaysia, he stuck to his idea of running Malaysia as a secular country where people of different beliefs and religion lived and worked together. An enthusiastic sportsman himself, Abdul Rahman promoted many sports events in Malaysia, viewing is a medium to bring people of different race and religion together. He started an international football tournament in 1957 and was elected as the first president of Asian Football Confederation in the next year in 1958. He had a passion for horse racing and was a member of the Selangor Turf Club. Later Life Death In 1977, Abdul Rahman became the chairman of The Star, a newspaper which was banned in 1987 by Prime Minister Mahathir Mohamad because of its provoking columns that were harshly critical of the Malaysia’s government. Following a rift with in UMNO, he unsuccessfully tried to establish a new party UMNO Malaysia. In the general election of 1900, Abdul Rahman actively participated and widely campaigned against Mahathir Mohamad despite his worsening health. He died on 6 December 1990, aged eighty seven and his body was buried at the Langgar Royal Mausoleum in Alor Star. Family Children Abdul Rahman is believed to have at least four marriages in which only three are officially confirmed. His first wife was Meriam Ching, a Chinese woman who gave birth to two children Tunku Khadijah and Tunku Ahmad Nerang. After her death, Abdul married Violet Coulson, his former landlady in England. He divorced her and married Sharifah Rodziah Syed Alwi Barakbah and the couple adopted four children Sulaiman, Mariam, Sharifah Hanizah and Faridah. His fourth marriage to a Chinese woman Bibi Chong remained secret with whom; he had two daughters Tunku Noor Hayati and Tunku Mastura.

An Investigation of Safety Practices in the Saudi Arabia Construction Research Paper - 1

An Investigation of Safety Practices in the Saudi Arabia Construction Industry - Research Paper Example accidents 17 2.6.2 Continuous safety practices by top management 19 2.6.3 Continuous safety practices by safety professionals 20 2.7 Measurement of safety performance 20 3. Project Plan, Methods and Techniques 24 3.1 Project Plan 24 3.2. Methods and Techniques 24 3.2.1. Sampling 24 3.2.2. Data Collection 25 3.2.3. Data Analysis 25 3.3. Skills and Experience 26 3.4. Time Scale 26 List of References 27 1. Introduction 1.1 Background Saudi Arabia has experienced rapid growth over the last few decades. In 2011, the GDP growth rate was 3.8% (TradingEconomics, 2011). This is roughly an average for the 21st century with lows having occurred during 2002-2003 and 2010 and highs during the mid 2000s. Exports at present are over 200,000 million SAR and over 90% of these are oil exports, which account for 75% of government revenue. The rapid growth is therefore mainly due to increases in the kingdom’s oil revenues. ... The construction industry engages in designing, constructing and maintaining unique buildings. In Saudi Arabia, its growth in recent years has been remarkable, especially due to increased foreign investment from international companies seeking to expand their businesses. According to Saudi Arabia’s Infrastructure Report for the second quarter of 2011, Saudi’s construction industry is set to grow by 4% in 2011, and it will continue to grow at this rate for the next four years (Roberts, 2011). In particular, during the period 2011 to 2015, Saudi Arabia’s residential building construction is set to grow at a rate of 7.2% (Saudi Entrepreneurship, 2011). The Saudi construction sector is the largest and fastest growing in the Gulf region. The government is completing the establishment of six economic cities; it has invested almost SR513.8 billion during the period October 2008 to April 2009, and it is expected to spend around SR1.5 trillion over the next half decade (U S-SA Business Council, 2010). However, the construction industry is also one of the most highly hazardous industries. Its unique nature, difficult work-site conditions, human behaviour, and poor safety management are the main causes of accidents (Koehn and Chih-Shing 1995, 261). Unfortunately, despite the construction evolution in Saudi Arabia, the work methods, equipment and procedures are still unsafe. As an example, in a recent incident in Riyadh in January 2011, it was reported that three workers were killed and eleven were left injured when scaffolding collapsed (Abbas, 2011). Several other highly publicised accidents at construction sites have also taken place in Saudi Arabia in recent years. Regulations do exist but many

Health Beliefs of Haitians Essay Example | Topics and Well Written Essays - 1000 words

Health Beliefs of Haitians - Essay Example Always when a Haitian becomes ill, the first line of care is home herbal remedies. These are often utilized for the prevention and treatment of cold, fever, and stomach aches. Traditional treatments are passed on from generation to generation. It is difficult for them to accept Western approaches such as vaccination and cancer screening as they feel the treatment may make them ill and they have no prevention against it. Haitians believe that pain affects the whole body system and because of that they are frequently not able to tell you where the pain comes from. They also may come to the physicians office and be very vague about what is wrong with them because they see everything as the same (Salisbury.edu), either natural or supernatural. All deformities are considered brought on by an evil spirit. Haitians who have a chronic illness are cared for by family and friends, they seldom go to a nursing home. If they get to return home they will pay the spirits back by having a Thanksgiving ritual. Haitians also believe that a wheelchair means they are very sick and they will misunderstand if it is offered. Haitians do not talk about organ donation, nor do they believe in organ transplant. Pre-natal care is not an illness so there are most likely no prenatal care visits and it may be very difficult. Practitioners Most of the lower class in Haiti believes in Voodoo and that comprises about 85% of the population. They do practice Christian beliefs at the same time. There, according to the Haitians is not only a visible world but a spiritual world. The spirits of the deceased make up both good and bad spirits. Usually when there is an illness the Haitian goes to see the Hougan who is able to be a conduit to the Loas and provide a cure. If the patient has visited the Hougan several times and is not better, they may be referred on to the physician (Miller, 2000). If a patient is in the hospital here in the United States, they may want to go back to Haiti to see a Hougan, especially is they are not getting better. Physicians within the community of practitioners to treat Haitians must do what they can to understand the cultural significance of many of these beliefs in order to get screening and preventive care done. Chronicity and Psychiatry The role of the supernatural is very much a part of Haitian society. They feel that chronic illness as well as psychiatric illness is caused by the supernatural. Depression, psychosis, inability to perform activities of daily living and academic underachievement may often be seen as a curse or a spell placed on them. They often feel that this happens because they did not work hard enough of did not do something they should have done (Astrid & Shiela, 2002) and didn't. They may feel they were lazy or that someone had a grudge and put a spell on them through an evil spirit. Patients who are chronically ill are usually cared for at home by family and friends: nursing homes are almost never used. It should be remembered also that offering a Haitian a wheelchair is the same as telling they are very ill and may not get better (Salisbury.edu) In actuality, the Haitians deal with many chronic illnesses due

Business Ethics Essay Example | Topics and Well Written Essays - 500 words - 1

Business Ethics - Essay Example He decides about his behavior in the light of the ethical discussions because he has learned it through a sales training program which was conducted in the past. He came to the understanding that if he commits himself to ethical behaviors, his subordinates would also be ethical within their undertakings. He received instructions in writing within the sales training program and this formed the basis for his comprehension. Mr. James’ company employs a proper code of ethics because he has set his eyes on achieving the very best by remaining clean and transparent within his dealings with the people under him as well as the customers who visit his chain of stores. He maintains a proper relationship with them at all times. He leads his team through difficult situations with consummate ease because he has got a wait and see approach which is based on sound ethical values and principles. Companies in this day and age do have written codes of ethics because it dictates the way the empl oyees should conduct themselves. The punishments or penalties imposed on them for violating the ethical domains include the likes of suspension and other legal procedures, which are managed by the human resources management department within these companies.

Anopheles Stephensi Tissue-restricted Expression

Anopheles Stephensi Tissue-restricted Expression Tissue-restricted expression and alternative splicing revealed by transcriptome profiling of Anopheles stephensi Sreelakshmi K. Sreenivasamurthy1,2, Anil Madugundu1,3, Arun Kumar Patil1,4,5, Gourav Dey1,2, Ajeet Kumar Mohanty6, Manish Kumar1,2, Krishna Patel1, Charles Wang7, Ashwani Kumar6, Akhilesh Pandey1,8,9,10,11, T. S. Keshava Prasad1,2,4,* Abstract The sequencing of Anopheles stephensi, a major malaria vector in Asia has led to increased research activity to understand the vectorial ability of this mosquito species. However, tissue-based gene expression profiles of the annotated genes remain to be understood. In this study, we summarize the transcriptomic profile of four important organs of a female imago Midgut, Malpighian tubules, Fat body and Ovary. We identified over 21,000 transcripts in total, from all the four tissues corresponding to about 12,000 gene loci. This study provides an account of the tissue-based expression profiles of majority of annotated transcripts in An. stephensi genome and alternative splicing in these tissues. Understanding of the transcript expression and gene function at the tissue level would immensely help in enhancing our knowledge of this important vector and decipher the putative role of these mosquito tissues, providing the basis of selection of candidates for future studies on vectorial abil ity. Keywords: Mosquito, RNA-seq, differential expression, lncRNAs Introduction Malaria remains as one of the most debilitating mosquito-borne diseases till date. According to WHO World Malaria Report in 2016, there were ~212 million malaria cases in the year 2015, resulting in an estimated death of about 429,000 individuals globally. Most of these cases (90%) is in the African region with about 7% incidence in South East Asia. About 50% of the Asian malaria incidence and deaths has been in India1. The number of deaths attributed to malaria in India has been reported to be higher than the WHO estimates2. However, the latest updates on the cases and deaths reported in India has been limited to the National Vector Borne Disease Control Programme (NVBDCP), according to which there has been about a million cases of malaria reported in the year 2014 [http://www.nvbdcp.gov.in/malaria3.html]. Out of the 41 different Anopheline species reported as significant vectors for transmission of human malaria, An. stephensi is an important vector in India and South Asia3,4. Bein g the major urban vector, it is second most prevalent in India. It has been reported all over the country except the north-eastern states of Sikkim, Arunachal Pradesh, Mizoram, Nagaland, Manipur and Tripura5. Sequencing of the Anopheles mosquito genomes have resulted in a spurt of activity in the study of Anopheles mosquitoes. PubMed search with the keyword Anopheles resulted in 14,576 publications, majority of which have been after the year 2000 as shown in the Figure 1A. Majority of the studies post-genome sequencing has been focussed towards understanding the role of various genes and development of numerous methods to regulate their expression.   The overall aim of the community is to embark on a feasible means to control the spread of infectious organisms either by controlling the vector/mosquito population or by curbing or reducing their vectorial ability. In this regards, numerous studies have already been performed on the recently sequenced malarial vectors6-9. However, most of the studies are focussed on previously studied molecules with very few studies focussing on new target molecules. This could probably be due to the lack of reliable data owing to incomplete genome as semblies and annotations in the identification of such targets. We have tried to bridge this gap with a huge effort of supplementing the current efforts using an integrated approach of utilizing proteomic and transcriptomic data in the genome annotation and assembly in an array of organisms through our previous studies10-12. Although transcriptomic data played a major role in refining the annotations and assembly of the genomes in the previous study, the tissue-based expression profiles were not focussed on. The tissue-based expression profiles of the identified transcripts are the focus of this study. Tissue-based expression profiling in An. stephensi has been limited to salivary glands13, ovaries14,15, testes16 and hemocytes17 with most the studies being done on whole mosquitoes14,18,19. However, even with the availability of transcriptome data from ovaries, there are several issues. The study was mainly focused on identification of transcripts expressed in developmental stages and is of low throughput14. The other tissue-based expression studies published including one cDNA-based study of the salivary glands and another being cDNA sequencing of the transcripts from hemocytes, both tissues that were not included in our study. The focus of our study is on the Midgut, ovary, Malpighian tubule and fat body of a female An. stephensi imago. These tissues, along with the salivary glands play a very important role in the blood meal digestion and thus important for the life cycle of the mosquito vector and plasmodium species. However, we restrict ourselves to understand the molecular dif ference between these mosquito tissues in the uninfected sugar-fed state of the mosquito which we believe will provide the much-needed basic understanding of the role played by these tissues. To this extent, we performed comparative and deep transcriptomic analysis of these four tissues. Materials and Methods RNA isolation and sequencing Adult female An. stephensi mosquitoes grown at the NIMR field station, Goa, were dissected to obtain midgut, Malpighian tubules, ovaries and fat body. These dissected tissues were stored in RNAlater to preserve the RNA quality till RNA extraction. The RNA isolation and sequencing was performed as described earlier10,11. Briefly, the RNA isolated using Qiagen miRNeasy kit was used for the preparation of indexed RNA-seq libraries using TruSeq RNA Sample Preparation Kit v3. The indexed and pooled libraries were sequenced on two lanes (as technical replicates) of Illumina HiScan SQ platform. Read alignment and transcript assembly The raw reads were processed for quality filtration to remove ambiguous bases present due to the sequencing errors at the 3 end of the reads. Base quality filter of >20 was considered as good. FastQC (Version 0.10.1) tool was used to determine the quality of the raw data and poor quality calls with Phred score An. stephensi genome build (ASTEI2) downloaded from VectorBase (https://www.vectorbase.org/) using HISAT (Version 2.1.0)20 aligner with the default parameters. HiSAT2 was supplied with known annotations and Gene Transfer File (GTF), AsteI2.2 from VectorBase. The alignment of reads from each lane for each tissue was carried out individually against the reference genome resulting in eight different Binary Alignment Map (BAM) files. The .bam files for each tissue were then merged to obtain merged .bam files, one for each tissue. The aligned reads were assembled against the AsteI2.2 gene annotations, as reference, using the StringTie (version 1.2.1) assembler21. Assembled transcrip ts were further quantified and annotated into known and novel categories using the gffcompare in StringTie package as described earlier22. To determine novel transcripts as a transcript GTF file and all the StringTie assemblies were merged using StringTie-merge option. Novel isoforms and intergenic transcripts were obtained by comparing the merged StringTie assemblies of all the four tissues to the annotated transcripts from VectorBase using gffcompare. Coding potential of the identified transcripts was predicted by the use of the Coding Potential Assessment Tool (CPAT)23. Transcripts which were >200 bp in length with a CPAT score threshold of Identification of differentially expressed genes across four tissues Merged GTF file from StringTie was annotated in to different classes of transcripts using gffcompare with respect to the VectorBase annotations. Expression levels of transcripts as determined by the StringTie assembler were compared across tissues. The expression information from individual lanes were used as technical replicates for each tissue. Differential expression was computed using Cuffdiff after normalizing the data across samples by calculating Fragments per Kilobase of exon per Million Fragments Mapped (FPKM)24. The R-package version 2.16.0 of cummeRbund was used for visualization, analysis of RNA-seq data and cluster generation25. An overview of the analysis pipeline is provided in Figure 1B. To identify tissue specific transcripts, we initially filtered transcripts with FPKM value à ¢Ã¢â‚¬ °Ã‚ ¥ 1.0 in at least one among the four tissue types. We then applied the right-tailed t-test to identify the transcripts which are relatively high in abundance in one tissue as again st other tissues. Results and Discussion Transcriptome sequencing of four An. stephensi tissues Midgut, Malpighian tubules, Fat body and Ovary was performed to create a tissue-based expression profile. In total, about 500 million paired-end reads of 100bp were generated from all the four tissues, with about 55 million read pairs per tissue sample from two lanes. The expression levels of transcripts between the replicates and among the tissues were comparable. Figure 2A represents the inter-tissue and intra tissue transcript expression variations in the form of a distance-based heatmap. The variations are minimal between the replicates as expected and increases between the tissues with Ovary and Malpighian tubules being the most different. By following the standard alignment and assembly pipeline using the HiSAT2 and StringTie assembler, we identified a total of about 25,000 transcripts. However, after the initial filtering for the FPKM values (à ¢Ã¢â‚¬ °Ã‚ ¥ 0.1) only 21,500 transcripts were retained. The expression of th ese transcripts was comparable across tissues with the median FPKM value ranging about 2 to 3 in all the tissues as represented by the box plot in Figure 2B. Figure 2C and 2D provides the general distribution of the length and the FPKM values of the transcript assemblies across the four tissues. About 60% of the transcript assemblies were found to have FPKM value of 1 and above, while the average length of majority of the transcripts tend to be in the range of 1000 to 3000 bp. This shows an expected trend of a reliable depth and absence of any skewing. The Transcript assemblies were classified into different classes using gffcompare. However, in order to avoid over interpretation of the data we have only focused our findings on the known =, alternate j and intergenic unknown u class of the transcript assemblies for our analysis. In our analysis, we noticed that almost equivalent number of transcript assemblies were classified under the known (=) and the alternate (j) categories. In fact, the transcript assemblies in the j category exceeded the number of known transcript assemblies. A deeper look in to this matter showed us that due to the poorly annotated gene models (which is mostly based on the prediction program) for this strain, the untranslated regions (UTRs) of the predicted transcript models in the current annotation is missed. As a result, the transcript assemblies with the extension of the exonic regions supported by the reads, probably into the UTRs were classified as alternate transcripts. We are working closely with the VectorBase to improve the annotations of these predicted gene and transcript models for the An. stephensi Indian strain. Tissue restricted transcripts Majority of the transcripts identified (about 87%) were expressed largely at similar levels in all the four tissues, the remaining 15% of the transcripts identified seemed to have more of a tissue restricted expression. Figure 3 details the distribution of the transcript expression (expressed with FPKM values à ¢Ã¢â‚¬ °Ã‚ ¥ 0.1) among the previously annotated transcripts (Figure 3A), alternative isoforms (Figure 3B) and novel previously unannotated intergenic transcripts (Figure 3C). The majority of the transcripts in each of these groups are expressed in all the four tissues with only about 3 4% of the transcripts showing tissue restricted expression. Among the known/annotated transcripts identified, 241 were found to be exclusive to Midgut, 221 exclusive to Malpighian tubules, 479 transcripts in Ovary and 436 in Fat body. The distribution of tissue specific transcripts was similar in the alternative isoforms and novel intergenic transcripts of these four tissues with 61, 67, 146 and 77 isoforms exclusively identified in Midgut, Malpighian tubules, Ovary and Fat body. In general, there was a clear bias in the number of transcripts and transcript isoforms that were common between midgut and Malpighian tubules and similarly between fat body and ovary than amongst the others. The diversity of the transcripts identified was found to be maximal in Ovary with most the transcripts being identified in this tissue, followed by fat body. Midgut had the minimal number of transcripts identified, however, the expression levels of these transcripts, in terms of FPKM, were higher than that of other tissues. Novel splice variants and their expression Apart from the known/annotated transcripts, we identified a plethora of spliced (exon-exon) reads that were not previously annotated. Assembly of such reads along with the intra exonic reads led to the identification of >8500 transcripts that were spliced differently. These alternatively spliced isoforms represent the complexity of the transcript forms and their expression in the four tissues. A summary of the differential expression of these alternate isoforms is provided in Figure 3B. As in the case of annotated transcripts, most of the alternatively spliced forms were also expressed in all the four tissues. Only about 1-2% of the total alternate transcripts isoforms were found to have tissue restricted expression. Transcript isoforms were enriched maximally in Ovaries compared to any other tissue. With 146 isoforms restricted to ovaries, it showed the highest variation in the spliced forms among the four tissues although the FPKM values for these were comparatively lower than that of other tissues. Fat body had the least representation of the alternate isoforms. The splice variants identified included examples of intron retention, alternative 3 or 5 donor and acceptor sites, exon skipping and others. Different spliced forms were expressed in different tissues. An example of transcript expressed in different tissues is provided in Figure 4. The annotated gene ASTEI04270 belongs to the Gelsolin/Vilin/fragmin superfamily, coding for a single transcript isoform according to the VectorBase annotation. However, we identified six different isoforms for the gene. The original protein coded by the annotated transcript with a signal peptide and nine gelsolin-like domains that was highly expressed in Fat body followed by Malpighian tubules. The alternative isoforms included a shorter transcript encoded by the first 3 exons (ANSTF.3986.4), which retained only three of the nine gelsolin-like domains along with the signal peptide sequence that was highly expressed in fat body and least expression in ovaries. The other 4 isoforms encoding the exons from fo urth exon consists of 4 gelsolin-like domains. Isoforms ANSTF.3986.1 and ANSTF.3986.2 were highly expressed in midgut followed by Malpighian tubules but not identified in fat body and ovaries. Whereas, isoforms ANSTF.3986.5 and ANSTF.3986.6 were significantly expressed only in midgut. Proteins encoded by this superfamily typically consists of three to six gelsolin-like domains (GEL), with each domain playing a critical role in actin filament remodeling26,27. Novel intergenic transcripts In addition to annotated and alternate spliced forms of the transcripts in the known/annotated gene loci, we found additional loci in the genome of An. stephensi Indian strain. The reads mapping to these unannotated regions were processed to assemble putative transcripts that were categorized as novel/unannotated transcripts. We identified about 2700 transcripts with FPKM values above 0.1 in the intergenic regions of the genome that were previously considered to be non-transcribed. The expression of most of these intergenic transcripts were found to be similar in all the four tissues. However, Expression-based clustering and functional correlation Since An. stephensi genome was recently sequenced and is relatively less worked upon, there is limited information on the function of these genes and transcripts. However, Gene Ontology analysis based on their translated protein and the domain structures (Interpro domains) showed that most of the differentially expressed transcripts were found to have expected domains as per the perceived function of these respective tissues. The identified transcripts were segregated into clusters based on their expression levels in the four mosquito tissues. Among the various clusters generated using the cummerbund package, few of the clusters showed clear trends of expression. One of the clusters with about 950 transcripts showed similar expression in all the four tissues. Gene level ontology mapping of these transcripts showed that majority of the transcripts possessed generic domains such as protein, nucleotide and ion binding domains, transmembrane transport, proteolysis, oxidoreductase activity and signal transduction (Figure 5A). Transcripts found to be enriched in the Midgut (170) compared to other tissues were found to have proteolytic, protein binding, hydrolase and peptidase activity. Some of the midgut enriched transcripts were found to be involved in chitin and carbohydrate metabolism (Figure 5B). Transcripts enriched in Malpighian tubules (116) were found to be associated largely with transmembrane transpor tation, oxidation-reduction process, protein and ion binding events. Few of the transcripts were associated with transferase, ligase and lyase activities among other catalytic activities (Figure 5C). Ovary enriched transcripts (241) were associated more with the protein binding, nucleic acid and ATP binding, in addition to those having signaling domains and transport domains associated with intracellular signal transduction processes such as GPCR activity, protein phosphorylation and dimerization. As expected, these transcripts seem to be involved highly in cell cycle processes including DNA replication, microtubule organization, DNA repair and growth factor activities, which are crucial mechanisms for vitellogenesis (Figure 5D). Fat body enriched transcripts (170) were consistent with the role of fat body akin to the vertebrate liver. The transcripts enriched in fat body are associated majorly with transmembrane transportation, oxidation-reduction process, chitin binding and metabo lism, heme-binding and transport, in addition to oxidoreductase activities (Figure 5E). Identification and expression of long non-coding RNAs We compared the list of transcripts identified in our study to the list of transcripts that are annotated as non-coding RNAs in VectorBase. However, we failed to identify any of the annotated non-coding RNAs in our study since the annotated ones are largely rRNAs and other small ncRNAs. Due to the ribosomal RNA depletion employed in our study, we expected no rRNAs to be identified. However, in order to investigate the presence and expression of the long non-coding RNAs in An. stephensi, we assessed the coding potential of all the identified transcripts using the CPAT tool. From this, we identified 4,071 transcripts that satisfied the criteria for the long non-coding RNAs (lncRNAs) (Supplementary Table 2). That is, they were longer than 200 bases in length and were predicted to have a coding potential of Tissues considered in this study play an important role in the life cycle of the female mosquito. They are critical in blood meal digestion, metabolism, vitellogenesis, excretion, immunogenesis, Plasmodium sporogony and reproduction, which are associated with vector physiology, progression and malaria transmission. Mosquito midgut is involved in the initial storage and digestion of the ingested blood. The gut epithelium also provides site for development of oocysts and sporozoites (Sporogony). Blood meal induces pathways such as TOR, which ultimately leads to synthesis of proteins required for egg development. Fat body and ovary are known to be involved in the utilization of the nutrients from blood to enable vitellogenesis. Malpighian tubules are known to play an important role in the mosquito xenobiotics. Fat body cells (trophoblasts) and recently, Malpighian tubules have also been shown to be involved in the immune responses28-31 and is now being considered as targets for mosquito control28,31. Towards this end, we further evaluated the expression of genes previously reported to be involved in the vector-pathogen interactions32 across the four tissues (Table 2). The affordability and accessibility of sequencing-based techniques have resulted in numerous transcriptome-based studies even in An. stephensi14,15,17,19. However, due to the low depth of the other existing studies, no significant comparison could be performed between the transcript expression from our study to that of the other studies. We deciphered the genes reported to be involved in immunity14 and evaluated the expression information for the annotated transcripts and the novel alternate isoforms across the tissues (Supplementary Table 4). Although, there has been a recent study of the cDNAs from hemocytes, we could not compare the genes expressed in their study since hemocytes were not part of our study. Another reason for non-comparison was normalization issues caused by 36bp single end reads in their study, with only 49% of it mapping to the VectorBase assembly. We provide the deepest tissue-based transcriptome profiling for these four organs of An. stephensi (Indian strain), so far. Studies such as ours depicting the transcript variations amongst tissues in its physiological states provide important baseline information. In light of such information, analysis of gene expression data in the context of changes due to blood meal, infection of insecticide resistance might lead to new perspectives and insights. This, in turn, will facilitate the choice of novel targets for vector control and transmission blocking studies and other experiments as evidenced in An. gambiae33. Data Availability The RNA-sequencing data has been submitted to the Sequence Read Archive (SRA) from NCBI and can be accessed using the project accession number SRP043489. Supplementary data Supplementary data are available at www.dnaresearch.oxfordjournal.org. Funding This paper is funded by the joint research project to NIMR and IOB entitled Characterization of Malaria Vector Anopheles stephensi Proteome and Transcriptome (EMR/2014/000444) from the Science and Engineering Research Board (SERB), Government of India. SKS and GD has been supported by the Senior Research Fellowship by University Grants Commission (UGC) and MK was supported by the Council of Scientific and Industrial Research, Government of India during the study. Table 1. Transcript distribution number of transcripts in total, class code-based classification of transcripts in all four tissues and in individual tissues      Ã‚   All 4 tissues Midgut Malpighian tubule Ovary Fat body Total number of transcripts identified 21,500 17,461 18,812 18,616 18,685 Corresponding gene location identified 12,256 10,357 11,107 10,973 11,371 Total number of known/annotated transcripts = 9,722 7,508 7,883 8,001 8,015 Number of alternate isoforms/transcripts j 8,820 7,603 8,232 7,992 8,037 Number of novel transcripts (intergenic) u 2,694 2,136 2,458 2,396 2,398 Figure Legends: Figure 1. A. Graphical representation of the remarkable increase in the number of studies on Anopheles mosquitoes post genomic era. B. Workflow representation of the study pipeline followed. Figure 2. Overall representation of transcript expression. A. HeatMap representation of the Jensen-Shannon (JS) divergence between the different tissues and their technical replicates. B. Bar-chart representation of the tissue-based transcripts and their median expression in the log10(FPKM), showing normalized distribution. C. FPKM distribution curve of the transcripts identified in the four tissues. D. Distribution of transcript length across the four tissues. Figure 3. Venn diagram representation depicting the overlap and the tissue specific expression of the transcripts across the four tissues A. For VectorBase annotated transcripts. B. Distribution of alternate isoforms of transcripts. C. Distribution of novel intergenic transcripts. Figure 4. An example representing the novel spliced forms of the VectorBase annotated gene ASTEI04270. Isoforms identified due various splicing events and their expression across the four tissues. Figure 5. Expression-based transcript clusters and the functional enrichment of the classes of transcripts based on domain and Gene Ontology-based functional annotation. A. Transcripts having similar expression in all four tissues B. Midgut-enriched transcripts C. Transcripts overexpressed in Malpighian tubules D. Transcripts highly expressed in Ovary E. Fat body-enriched transcripts.

Marketing Strategy for Marks and Spencer Food Essay -- AVCE Business S

This assignment is about marketing, where I will produce a marketing strategy for a new or existing product. This unit introduces the major principle and functions of marketing; I will look on customer needs, and creating a suitable strategy or marketing mix, which will satisfy customer needs. In this unit I will experience the marketing process from carrying out initial research about a market, investigating the principles of functions of marketing and the way in which it contributes generating income/profit in a business. Marketing is the process of planning and executing the conception, pricing, promotion, and distribution of ideas, goods, and services to create exchanges that satisfy individual and organizational goals. Marketing also involves analyzing customer needs, securing information needed to design and produce goods or services that match buyer expectations, and creating and maintaining relationships with customers and suppliers. Marketing is essential to the success of any business. Its primary aim is to enable businesses to meet the needs of their customers and potential customers, whether then it’s for profit or not. To make my strategy successful for business it must: Ø Understand customer needs Ø Understand and keep ahead of competition Ø Communicate effectively with its customers to satisfy customer expectations. Ø Coordinate its functions to achieve marketing aims Ø Be aware of constraints on marketing activities I will also be considering the importance of developing and maintaining a relationship with its customers, potential customers and other stakeholders. I will look upon on establishing customer needs, methods of analysing marketing opportunities, and the m... ...ons can exert a major influence upon the stores’ accessibility and attractiveness to consumers. Planning the selection process as carefully as possible and using methods described should maximise the success. Marks and Spencer pays particular attention to social and geodemographics, along with consumer spend, before deciding on a location. Own brands are desirable products to offer, as they offer profitable margins and a high level of control over the marketing aspects of management. Efficient use of these products can increase store loyalty. As I did a oral presentation I used few pictures and such to show the packaging design, as I talked about the other product and the company, and mainly on one of the marketing mix ‘the product’. My presentation could have been bit longer with more details and also posters to show big images then small ones.