Treatment Plan Exercise

Client Name: _________Yang_________________________________  Date of Birth: ______N/A________

Gender: _Female___ Age: ___41 years__________           Race: _____Brown________ Client Ethnicity: ______Chinese______________

Reason for referral for counseling:  Yang presented symptoms of posttraumatic stress disorder (PTSD) following the death of her son, Daiyang in the earthquake, as well as, the subsequent pregnancy, months after her son’s death. Other reasons may include;

·         She reported frequent flashbacks and disturbing memories of the scene of her son’s death that made her feel anxious and miserable

·         She was unwilling to talk about the earthquake

·         Yang reported significant sleep difficulties, memory difficulties, and anhedonia

·         After getting pregnant again, Yang reported nausea and loss of appetite

·         She reported problems in interacting along with her husband and neighbors

Concern 1: Disturbing memories as shown by the frequent flashbacks and nightmares related to the scene of her son’s death outside the building

Goal for improvement: Disturbing memories and symptoms of PTSD will be significantly reduced and they will no longer interfere with Yang’s functioning.

Objective 1: Yang will identify the issues that trigger the bad memories about her son’s death

Objective 2: She will be involved in an additional physical activity or sport

Objective 3: Yang will learn coping skills necessary for emotional regulations, for example, training herself not to think much about her son’s death

Concern 2: Avoidance to think about the earthquake as shown by Yang’s tendency to avoid news related to the Chinese earthquake (Leahy, Holland & McGinn, 2012)

Goal for improvement: Increasing contact with people, places and things that bring back the memories of her son’s death

Objective 1: Yang will list down the places, people or things that remind her about the earthquake

Objective 2: She will avoid staying isolated in her house and try to maintain contact with these places or people

Objective 3: Yang will be taught that turning to drugs or alcohol is a wrong way of reducing the pain and she should avoid drugs entirely

Concern 3: Physical stress as indicated by difficulties in sleeping, anger, anxiety, and conflicts with her husband (Leahy, Holland & McGinn, 2012)

Goal for improvement: Reestablishing a normal mood by becoming happy and confident, as well as, having enough sleep

                        Objective 1: Yang will be sleeping for 7 hours in a day

Objective 2: Yang and her husband will establish a timetable of family activities so that she can build a positive relationship with her spouse

Objective 3: She will start gaining mental strength by not dwelling much on the past, but focusing on the future

Primary Diagnosis: Stressor: The person was exposed to a death scene (APA, 2013).

Secondary Diagnosis:  Intrusion symptoms were evident as the client persistently re-experienced the traumatic event (APA, 2013).

Additional Diagnosis: Avoidance: The patient showed persistent effort to avoid thinking about the traumatic event (APA, 2013).

Reasons for choosing the goals and objectives

Concern 1

Goal: Yang needs to resolve the troublesome feelings that come with the memories of her son’s death in order to ease the depression

Objectives: I chose the objectives because Yang needs thought stopping and switching skills, as well as, progressive muscle relaxation to gradually stop the disturbing memories

Concern 2

Goal: When Yang increases contact with the things that trigger the memories, she will learn how to switch thoughts about the earthquake whenever they come.

Objectives: A step-by-step plan is necessary to ensure Yang knows the triggers of bad memories, avoids lonely places and keeps drugs away from herself

Concern 3

Goal: When Yang becomes happy, she will relieve the general anxiety that is caused by frequent flashbacks. In this way, she can reduce the sleep disturbances at night.

Objectives: Setting a target of sleeping hours, as well as, developing a timetable of family activities will keep Yang always committed to actions that will eventually make her happy and less anxious.

Reasons for choosing the diagnoses

Primary Diagnosis: Yang saw the scene where her son’s body was laid outside the building.

Secondary Diagnosis: Yang had nightmares, flashbacks and disturbing memories about the earthquake

Additional Diagnosis: Yang avoided news related to the earthquake and refused to talk about it entirely

References

Leahy, R. L., Holland, S. J., & McGinn, L. K. (2012). Treatment plans and interventions for depression and anxiety disorders. New York: Guilford Press. Obtained from: http://www.cognitivetherapynyc.com/leahch06.pdf on June 17, 2017.

American Psychiatric Association (APA). (2013) Diagnostic and statistical manual of mental disorders, Washington DC: APA. Retrieved from: http://creativityandmadness.com/wp-content/uploads/2014/07/Wilson-Handout.pdf on June 17, 2017.

How Can the Society Address Climate Change?

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How Can the Society Address Climate Change?

            In the current, evolving world, the issue of sustainability now lends itself as one of the critical issues more than ever. It is arguably one of the outstanding subjects that policymakers are discussing. This issue is hinged on the view that the growing global populations, the high rates of depletion of natural resources and the various emerging social, health and economic challenges call for a rethink on ways that the current generation will be able to continue sustaining itself, considering the effectiveness of existing approaches have been questioned. Many of the discussions have acknowledged the role of the environment as particularly imperative to sustainable development. The primary premise for this position is that if the global community does not protect or conserve the environment, the adverse environmental challenges such as famine, natural calamities, and diseases will be experienced, and these results will subvert the efforts aimed at achieving the social and economic development goals (Gille, 5). However, the path to environmental sustainability has not seemed to be a straightforward one. Indeed, several views have been offered as strategies for sustainable development, some of which have elicited the questions concerning their appropriateness. Some suggestions have always been characterized by heated debate contests. In fall 2008, the head of the United Nations Intergovernmental Panel on Climate Change, Rajendra Pachauri, called upon the global society to eat less meat in a bid to conserve the environment. His view has been perhaps the most interesting of insights that have attracted sharp reactions. This paper explores the question of eating meat as a way of supporting environmental protection, focusing on the implications of Hamilton's argument on Dr. Pachauris viewpoint.

A Look at Dr. Pachauris Opposing Viewpoint

            Dr. Pachauris acknowledges that the state of the rising global temperatures cannot be ignored — it calls for drastic interventions. The most appropriate approach for this intervention is by narrowing on some of the anthropogenic activities responsible for the emission of most of the greenhouse gasses. Animal farming happens to be one of the most notable causes of greenhouse gasses and, therefore, one way of addressing the issue is by avoiding eating meat. Dr. Pachauris viewpoint rests on the startling statistics that the meat production processes account for about 25 percent of the greenhouse gas volume emitted from the globe. These amounts of pollutant gasses are produced during processing of animals feeds, while others, especially methane, are emitted from ruminant digestion, and this happens to be about 23 times more effective in contributing to global warming than carbon dioxide.

            The essence of reducing consumption of meat is to lessen the meat demand, which would translate to reduced animal farming activities. Dr. Pachauris concerns are expressed at the backdrop of the growing demand for meat, which is projected to double in the next 5 decades. Therefore, reducing consumption of meat would be the only rapid, feasible approach that would enable the global community to tackle the worsening global warming issue. Dr. Pachauris advises the regular meat consumers to give up taking meat for one day a week, and then continue cutting down the consumption more and more. Apart from reducing meat consumption, Dr. Pachauris has advised that the strategies to tackle climate change will need to be accompanied by other forms of lifestyle change, which will help reduce the amount of gas emissions, and this should be reflected in different sectors.

A Look at Criticism

            The approach has attracted criticism from various individuals, for instance, Lisa Hamilton. Lisa Hamilton provides a relatively different approach to environmental sustainability — rather than eat less meat as Dr. Pachauris urges, she considers that people should, in fact, eat more meat. Her views are essentially borne on two premises; livestock is a critical component of the ecosystem protection equation and are a source of revenue and food.

            First, while Hamilton does not refute livestock accounts for a significant percentage of greenhouse gasses, she suggests that deciding not to have them will still not address the issue of methane produced by other organisms such as deer and terminates. If only, livestock is important because it produces manure that can be used to enhance soil fertility. In essence, farmers will not be able to generate high yields in the absence of livestock. According to her, the best approach to dealing with carbon problem is to get it back to the soil, and that happens only when the livestock is in the conservation equation. Moreover, cattle play a much more crucial role other than keeping the soil fertile —when managed properly; they can enrich the capacity of in sequestering carbon. Therefore, cattle do not only add manure to the ground, but their symbiotic grazing also encourages the growth of plants, while their hooves crush plant residue to support the ecosystem needs. Secondly, if cattle are well managed, intensive grazing processes may play a crucial role in shifting the emitted carbon to so significant levels that livestock farming can be perceived as a way of controlling the menace of greenhouse gasses. To her, the potential of animal benefits is yet to be realized because the meat consumption is still low.

            Therefore, for Lisa M. Hamilton, it is plausible to encourage people to eat more meat and promote livestock farming activities because they are not only an economically viable project but also an environmentally friendly event.

Reflection

            A look at Dr. Pachauris and Lisa Hamilton insights reveals opposing viewpoints. While both do not refute the need for environmental conservation and the presence of high percentage of greenhouse gasses emitted by livestock, they hold a different perspective of what must be done about cattle. In particular, based on Lisa Hamilton’s perspective, Dr. Pachauris approach can be conceived as a radical one — it overlooks the economic and ecological benefits that livestock offer. She sees that if the livestock is not reared, farmers will not be able to earn a good living, and at the same time, other derivative benefits such as manure will be lost.

However, it is noteworthy that, other than defending the economic and a few ecological benefits of livestock, even Lisa Hamilton’s perspective does not provide a succinct explanation on how the society could deal with associated greenhouse emissions. Rather, her response is a thesis that it is not just the livestock that is involved in the production of greenhouse gasses, but also other organisms such as deer and termites. Therefore, her perspective is critical that reducing livestock may not have much to do in reducing greenhouse gas emissions. While she posits the possibility of how livestock could be well managed to cut down greenhouse emissions, her insights do not adequately specify how this could be done, as well as the rate the livestock management approaches can subdue the actual greenhouse gasses the farms emit.

Conclusion

In conclusion, the aim of this paper has been to reflect on the issue of eating meat for the environmental protection, focusing on the implications of Hamilton's argument on Dr. Pachauris viewpoint. On one hand, Dr. Pachauris acknowledges that the state of the rising global temperatures cannot be ignored — it calls for drastic interventions, and that since livestock farming happens to be one of the most common causes of greenhouse gasses, one way of addressing the issue is by avoiding eating meat. On the other hand, Lisa Hamilton provides a relatively different approach to environmental sustainability — rather than eat less meat, people should, in fact, eat more to conserve the environment, reasoning that livestock is a critical component of the ecosystem equation, and are a source of revenue and food, too. Therefore, Hamilton's and Dr. Pachauris viewpoints can be seen as opposites. Based on Lisa Hamilton’s perspective, Dr. Pachauris approach to the issue is only a radical one that overlooks the economic and ecological benefits that livestock offer. She sees that if the livestock is not reared, farmers will not be able to derive a living, and at the same time, other derivative benefits such as manure will be lost. While Lisa Hamilton’s perspective does not provide a succinct explanation on how the society could deal with the associated greenhouse emissions, she nevertheless provides alternative thinking to the issue, which certainly weakens Dr. Pachauris viewpoint. Such points of view, however, reflects the inherent challenges regarding the path to protecting the environment.

Works Cited

 Gille, Sarah "Warming of the Southern Ocean Since the 1950s". Science. 295 (2012): 1275–7. Print

Laboratory Report on Gene Cloning into Expression Vector

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Laboratory Report on Gene Cloning into Expression Vector

Abstract

Gene cloning involves manipulation of a given cellular component to isolate the desired gene properties. Isolation of particular cellular component forms the core of the cell chemistry and biology. For a gene to be efficiently cloned, it must be inculcated in a larger medium that will allow sufficient expression of its markers. The vectors form the structures viable enough to encourage the manipulation of the genes to get the desired efficacy. Mostly used vectors are the plasmid vectors. They are ideal because of their neutrality nature and the growth favoring characteristics. The cloning vector obtained from the plasmid is useful and provides a good binding for the foreign DNA fragments which allows for the elucidation of cloned gene. This laboratory report explores on how gene cloning is altered into an expression vector. It also highlights on the Polymerase Chain Reaction and how the process is used to amplify the cloned genes into the desired particles. The last section explains the findings of the experiment.

Keywords: cloning vectors

                    Polymerase Chain Reaction

Introduction

For any desired effect of gene cloning to have an impact, there must be a proper layout on the ways to elucidate the genes properly. The use of expression vector is effective method since the vectors provide the necessary requirements and conducive media for an insertion of foreign DNA genes. In the experiment, tomato cells were used as the expression vector to plant with the gene of interest. Ideally, for a successful experiment to be conducted on the gene cloning, different sets of parameters must be taken into consideration. First, the gene has to be isolated. Secondly, the gene has to be cloned into a vector for a greater expression, and lastly, the gene must be expressed and translated to give the desired aims of the experiment (Brown 10). The laboratory experiment conducted followed the same fashion. Adjustments were made in the vector to meet the typical qualities of eukaryotic cells. The first step taken into consideration was the use of promoter sections, Kozak sequence and inserting appropriate start and stop codons. In addition to the requirements, kanamycin resistance gene was used as selected marker to identify the cells that have taken up the DNA.

Materials and Reagents Used

Isolated cDNA gene

Plasmid, pXCN

Escherichia coli strain

5microlitres buffer

3microlitres MgCl2

1microlitre of dATP

40microlitres purified PCR Product

1microliutre Taq DNA polymerase

Incubator

Ice cubes

Test tubes

Method

The above reagents were added in a sequential manner while observing the time intervals to realize the efficiently purified product.

One purification was done, transformation protocol followed.

The competent cells were left for some time to thaw on the ice.

5microlitres of litigation mix was then added with a lot of caution to the competent cells and the mixture left for further 10 minutes on the ice.

The tubes were then put into a floating tube holder and placed in a 42 degrees Celsius water bath for 1 minute to achieve acclimatization temperatures.

The cells were immediately put back on ice and kept for further 2 minutes

After that, 1milliltre of LB was introduced to the mixture and subjected to incubation for 60 minutes at 37 degrees Celsius.

On completion of the incubation, the cells were spun to concentrate them on LB plus kanamycin.

Week 2

Screening the Plasmids

This part of the experiment was mainly aimed at screening the vector to identify if the DNA fragments are correctly inserted as desired. The technique used was Polymerase Chain Reaction.  The bacteria colonies formed were screened to make sure that the plasmids were in line with the size of wanted genes and the standard ranges.

Method

Each bench set up a 4times PCR master mix. The required master mix estimated for the experiments was 100 microliters.

The master mix used was kept on the ice to maintain the ambient temperature.

The ingredients were added in the following proportions; 2.5ul and 25ul of 10x buffer were added to 1x volume and 4xMaster mix of 25ul and 100ul respectively. Magnesium Chloride was added to the same mixtures starting from a volume of 0.75ul and 3ul. Primer mix was then added to the same mixture starting with 0.5ul and 2ul to the different master mix volumes. dNTPs were added in the same pattern but in different volumes of 0.5 and 2ul. Water ice then followed the addition sequence in the order of 20.15ul and 80.6ul. Lastly, Taq Polymerase was added starting with a volume of 0.125ul and 0.5ul to the respective master mix volumes.

After the additions, the solutions were gently mixed using 80ul set pipette avoiding bubbles.

25ul of the solution was then transferred into the PCR tube and the DNA template from part of the bacterial colony taken. The samples; were then set to run in PCR machine using standard conditions. The denaturation, annealing and elongation temperatures were set as 94, 58 and 72 degrees Celsius respectively with following timings per cycles made.

After the PCR run, the product was loaded onto the agarose gel and results analyzed.

Results

PCR Screen 1

Clear bands formed

Proper orientation of the base pairs

Clear visibility of PCR wells

Diagram

PCR Screen 1

PCR Screen 2

No clear bands on the formation of the required primers. 1000kb primer is not expressed in the sample loaded in the gel agarose. Additionally, there is an irregular arrangement of the base pairs.

Diagram

PCR Screen 2

Restriction Digest Screen

There is obstruction of the primers hence no clear bands are observed. The base pairs are irregular placed. The bands formed do not coincide with the required base pair primer expected in the screening results.

Diagram

Restriction Digest Screen

Blast Analysis

Blast X was used to find the nucleotide query sequence against protein database sequence. The nucleotide sequence used covered the entire mRNA with proper alignment of the base pairs to form stop codon, TGA.

Electrophoretogram

It is a tool used in analyzing the presence of nucleotide sequences by passing it through electric current. The number of wavelengths depicts the presence of nucleotide sequence. In the experiment, cytosine expressed greater wavelength followed by Adenine and subsequent base pairs. The pattern displayed movement of the charged particles in the agarose gel.

Discussion

The PCR Screen 1 was most efficient among the other experimental methods used. It presented a clear outline of the band coinciding with the required primer one. The insertion of the plasmid evidenced by the presence of clear band indicates that there is the proper introduction of the forward primer and reverse primer having points of origin. In restriction digest screen, the presence of BamHI restriction enzyme although tend to be valuable in promoting the insertion of the primers into the cell, the exact area of cutting cannot be estimated therefore raises a discrepancy on the formation of the actual band (Davis 12). When BamHI cuts the cell, it creates a surface for appropriate insertion of both the reverse and forward primers. The method is not effective because the precise cutting point is not known.

PCR Screen 2 defines how the recombinant plasmid orientation is formed. The band formed is not clear, a likely explanation on the random combination of annealed forward and reverse primers. A clear outline is thus not shown in the method. The orientation of the gene is not inserted correctly. The sequence results of the restriction digestive show a messy situation with irregular peaks. This is also in line with screen two results.

The Recombinant plasmid screen displays the mixed result based on the various performances of the three screens. In the display, the variation is tied to inconsistencies realized on the second screen and the restriction digestive screens. Therefore, no clear markings are shown in the recombinant blot. The sizes vary and present smears instead of bright bands.

Precise and efficient gene cloning into vector expression requires a combination of different determinants in which each plays a significant role in the insertion of the primers. A good backup for that is the point of origin of the annealed forward and reverse primers (Unger 34). The two must face each other to form a synergy at the cutting points and create an active formation of the plasmid.

The process of gene cloning requires the manipulation of cells to meet up the behavior characteristics of a Eukaryotic cell. The experiment took into account the use of promoters to enhance the binding of DNA and its expression. The cells were also manipulated to adopt a Kozak sequence where the start and stop codons shift in the operations (Davis 20). The first phase of the experiment involved ligation reactions induced in Escherichia coli because it does not have the antidote. Appropriate follow-up of the gene coding should be followed to minimize chances of the discrepancy. In most of this case scenarios, Sanger Sequencing reactions should be used in this cases to check where abnormal ddNTPs have been added along the sequence.

The experiment ends with the formation of cDNA library. It begins with placing mRNA in a test tube. PolyT oligonucleotide is then mixed with the sample (mRNA) in the tube. The oligonucleotide will bind to the 3^’ end of mRNA which has polyA tail. The synthesis of cDNA library from mRNA needs an action of reverse transcriptase which alters the replication dogma enhancing the formation of the cDNA (Unger 35). RNAse is added to eliminate the RNA strands which will form short fragments that can later be used as primers in the formation of another DNA strand.

Diagram of the principle of cDNA library Formation (Unger 36)

In conclusion, gene cloning is an ongoing biomedical innovation that serves as a vital link in the manipulation of cells to give the desired effect. The cloning of the cells to provide the vector you want to require an excellent analysis of the orientation of forward and reverse primers, antibody resistant gene e.g.  Kanamycin and Tetracycline among others. The experiment explicitly covered on gene cloning into an expression vector.

Works Cited

Bio4442017 Laboratory Practical Manual

Brown, Terence A. Gene cloning and DNA analysis: an introduction. John Wiley & Sons, 2016: 10-63

Davis, Leonard. Basic methods in molecular biology. Elsevier, 2012: 7-58

Unger, Tamar"Applications of the Restriction Free (RF) cloning procedure for molecular manipulations and protein expression." Journal of structural biology 172.1 (2010): 34-44.