epithelial sodium channel (ENaC) in the human kidney.

Analysis of the data was based on two sample groups of individuals. These samples included healthy people and hydroxylase deficiency, people. RT-PCR method was used for the project, with the utilization of six primers for the epithelial sodium channel (ENaC) in the human kidney. The data analysis was done to depict the expression of each gene on human. The results are typically normalized to the gene of beta subunit with the exception of the housekeeping gene. The need to use urine samples from the two groups of individuals was to determine the expression of each gene on humans, from the perspective of such samples. To get perfect results, the urine donors had to be free of acute illness and were supposed to be free of any medication during the sampling.

Typically the ENaC has been found to be a heteromeric channel that has three similar subunits that are distinct. These include alpha (α), beta (β), and delta (γ).[i] These three subunits are regulated developmentally. They are also controlled differently by aldosterone. The alpha subunit, in particular, and its variants are expressed within the epithelial cells in the lungs. They are typically suppressed by oxidative stress.

To study the DNA synthesis, for all the subunits, each respective primer had to be studied. This is critical because primers are the basic points for DNA synthesis.  The 669-alpha primer consists of both forward and reverses primers on its 12th exon. All alpha primers have 12 exons. The 728-alpha primer has the reverse and forwards primers on is 1st exon. The two primers are shown in figure one and figure 2.

 Figure 1: a669- ENaC Primer

Figure 2: a728- ENaC Primer

The mutations of the beta subunit, specifically its last exon, have been found to have the potential of leading to Liddle’s syndrome.  Nevertheless, the forward primer process has been noticed between the 4th and 5th exon while the reverse primer process has been noticed in the 5th exon as shown in figure 3 below:

Figure 3: Figure 2:  ENaC Beta Primer

In essence, the gain of function mutations in the beta and delta subunits of the epithelial sodium channel has been found to contribute to pseudoaldosteronism, which is also referred to as the Liddle’s syndrome.[ii] Liddle’s syndrome is typically a form of severe salt-sensitive hypertension. It is considered to be a rare and monogenic type of hypertension.

Relatively, the delta primer has more exons than both alpha and beta primers. For the delta subunit, some difference was noticed as well. The forward primer process was noticed between the 5th and 6th exons while the reverse primer process was found to occur in exon 6 as seen in figure 4 below.

Figure 4: Figure 2:  ENaC Delta Primer


Other than the various primers, variations are also seen in the respective DNA gels. The alpha-669 ENaC had its DNA gel record a blood pressure of 94bp. This blood pressure is relatively normal. The DNA gel is as shown in figure 5.

Figure 5: Alpha-669 DNA Gel

The alpha-728 DNA gel recorded blood pressure of 150bp. This blood pressure depicts stage 1 of hypertension, which means that the patients’ blood pressure is higher than normal.

Figure 6: Alpha-728 DNA Gel

The cases of delta and gamma DNA Gels saw the subjects’ blood pressure level at 112bp for delta-638/74bp for delta-802 and 136bp respectively. This blood pressure is relatively normal, although it can be associated with medical problems like dizziness especially when subjects are standing. The results are seen in figure 7, figure 8 and figure 9 below.

Figure 7: Delta-638 DNA Gel

Figure 8: Delta-802 DNA Gel

Figure 9: Gamma DNA Gel


The study mainly focused on the beta subunit of the ENaC and the implication of its changes on hypertension.[iii] In this case, the beta-DNA gel was observed to be portraying a normal blood pressure since 77bp was recorded.

Figure 10: DNA gel for Beta ENaC

Generally, the use of DNA gel was to show the potentiality of hypertention with respect to the recorded blood pressure.

To establish the relationship between the beta ENaC subunit in the human kidney and its relation to hypertention, a copy number analysis was used for the DNA copy number variation for the samples. The copy number of a DNA sequence refers to the number of copies of such DNA a genome region. The main aim was to detect the variations, which could cause hypertention. Generally, genetic diseases that have Mendelian transmissions are characterized by some mutations that have rare allelic frequency. A small human population of about 0.1% is usually affected by the associated severe phenotype.[iv] More frequent mutations that are often of single nucleotide polymorphisms (SNPs) are found to have allelic frequencies of 1% or even 2% within the human populations. Other polymorphisms like copy number variations have been considered in this study. From previous studies, it has been found that genetic aspects account for 30% of blood pressure variation in human population. It can thus be asserted that hypertention development depends on the interaction between risk factors and genetic factors. Many genes are found to be associated with the control of blood pressure or essential hypertention.[v]

Regarding the beta gene to which the results were normalized, there are two common beta ENaC variants. These include the G589S and the novel intronic i12-17CT substitution. From the data analysis, it was found that the beta gene was slightly higher among the 21 hydroxylase deficiency group as compared to people within the healthy group based on the copy number values for each case. The β hydroxylase group of people was found to experience a sharp drop in their blood pressure.[vi] The higher value is attributed to the fact that these individuals are already suffering to hypertention. Regarding the same issue, mutation within the DBH gene results to the hydroxylase deficiency. Healthy people do not face the same mutation and thus they are likely to face a smaller effect of the mutation. A general copy number analysis of the β-ENaC showed that most of the subjects were below 50. Only nine subjects have recorded a copy number value above 50. Nevertheless, the two groups seem to depict a close trend in the variations of the beta gene. The results were presented graphically as follows:

Figure 11: Copy number analysis for healthy people

Figure 12: Copy number analysis for 21 hydroxylase deficiency people


Figure 13: Copy number analysis for Beta-ENaC

Further, the amplification of cDNA that is generated from 1ug of human RNA was done for each primer. This was done to measure the expression profile of the beta gene in comparison with other primers. The amplification results for the primers were as shown in figure 4 to figure 11 below.

In the amplification of cDNA generated from 1ug of human RNA, the aim was to show the relationship between the number of PCR cycle and yield in relative fluorescence units (RFU). This test depicted varying patterns of the relationship for every primer between the healthy group and the hydroxylase group of people.


Figure 14: Amplification of cDNA generated from 1 ug of human RNA


In the case of d802-ENaC, it seems that the PCR cycle number does not affect the yield in terms of RFU in the two groups. The effect makes a turn after the 30th cycle. In cycles 1 to 30, zero PCR volume is obtained. The two groups, however, show a different rate of change. The groups, however, appear to show an equal relationship between the PCR cycle number and RFU yield. Nevertheless, only 900 μl were used, which does not provide the chance to monitor beyond the 48th cycle. The two groups have their melt peak at 86 Degrees Celsius. Their RFU yield is however different at their melt peak as shown below

Figure 15: Melt curve analysis of above amplification

For a728- ENaC the trend is different for the two groups. It also turns out that the RFU yield starts at cycle 30. The rate is different for the two groups, but become the same at cycle 36. After cycle 38, the difference between the two groups is infinitely increasing as shown below.

Figure 16: Amplification of cDNA generated from 1 ug of human RNA

a728- ENaC

Figure 17: Melt curve analysis of the above amplification


Figure 18: Amplification of cDNA generated from 1 mg of human RNA for the beta gene

From figure 8 above, the melt curves (for healthy and hydroxylase deficiency groups) are leveled at zero for the RFU value until the 27th cycle. The two groups then depict a change in RFU at different rates. At RFU of 650 and at the 38th cycle, the two groups are showing a similar rate. The two groups did not seem to attain a maximum RFU even at the 50th cycle. Nevertheless, a melt curve analysis of the above amplification depicted different results as shown in figure 5 below.

Figure 19: Melt Curve analysis of the above amplification.

Figure 5 shows a melt curve for the two groups, but of the above amplification. The maximum RFU is reached at a temperature of 80o Celsius. The RFU then drops at a high rate until it levels at zero RFU after a temperature of 83 o Celsius. The amplification of cDNA was mainly done to obtain the length of RNA transcripts within the cells.[vii] The production of a cDNA copy of the required RNA sequence was successfully achieved.

The amplification of cDNA is a bit different. The RFU yield starts at cycle 24. This is far much lower than in both beta and alpha subunits. The difference in the curves keeps on widening as the yield volume and cycle number increase. This happens as depicted in figure 10 below.

Figure 20: Amplification of cDNA generated from 1 ug of human RNA for g- ENaC

Figure 21: Melt Curve analysis of above amplification.

Figure 11 shows a melt curve for the two groups, but of the above amplification. The maximum RFU is reached at a temperature of 82.5o Celsius. The RFU then drops at a high rate until it levels at zero RFU after a temperature of about 85.5 o Celsius. The amplification of cDNA was mainly done to obtain the length of RNA transcripts within the cells.[viii] The production of a cDNA copy of the required RNA sequence was successfully achieved.

Further, based on the data analysis, it can be established that the beta channel is a non-voltage and sensitive ion channel that is permeable and inhibited by the diuretic amiloride. The beta channel mediates the electrodiffusion of luminal sodium, including water that follows osmotically. This happens through the epithelial cells’ apical membrane. The channel is found to play a critical role in the pressure homeostasis of both blood and electrolyte. The beta channel also provides immense help in the liquid homeostasis of the airway surface. This aspect is important because it offers a proper mucus clearance. The channel also helps in controlling the re-absorption of Na in the human kidney, the colon, the lungs, as well as in the sweat glands. Finally, it plays a major role in the perception of taste.[ix]

Generally, the channel is end-effector within the rennin-angiotensin-aldosterone system. It resides within the apical plasma membrane. The apical plasma membrane is found in the renal cortical collecting duct. Re-absorption of sodium ions through the ENaC happens in collecting ducts of the renal cortical. The re-absorption is typically the final step for the sodium ion balance regarding renal adjustment. The ENaC is found to be playing an important role regarding the modulation of Na+ homeostasis thereby attributing the function to chronic blood pressure.[x]

From the figures depicting the results of data analysis, it is clear that the two groups did not show sharp differences in their expression of the ENaC beta subunit. While the group of people with the disease was used as a control, their results from their samples made it clear that hypertension was not solely related to the ENaC beta subunit. The rate at which the variations of the beta gene occurred among the two groups was almost similar. It can thus be asserted that hypertension is caused by a number of factors, besides the effects of the beta gene of the ENaC.[xi] Besides, it was previously mentioned that hypertension comes as a result of many factors. The factors are both hereditary and risk-related. The aspect of heredity reflects the effects occurring from the mutation of the respective genes. Since the beta gene seems to be equally occurring in people within the two groups, it is difficult to relate it to hypertension without considering other genes and risk factors as well.

[i] Basson, Jacob, Jeannette Simon, and D. C. Rao. “Between Candidate Genes and Whole Genomes: Time for Alternative Approaches in Blood Pressure Genetics.” Curr Hypertens Rep, vol.14, 2012: 46–61.

[ii] Tamura, H, et al. “Liddle disease caused by a missense mutation of the beta subunit of the epithelial sodium channel gene.” J Clin Invest, Vol. 97, 1996: 1780-1784.

[iii] Kawahara, Katsumasa, and Kouju Kamata. “The kidney and hypertension.” Clin Exp Nephrol, vol.16, 2012: 3–7.

[iv] Kawahara, Katsumasa, and Kouju Kamata. “The kidney and hypertension.” Clin Exp Nephrol, vol.16, 2012: 3–7.

[v] Kawahara, Katsumasa, and Kouju Kamata. “The kidney and hypertension.” Clin Exp Nephrol, vol.16, 2012: 3–7.

[vi] Persu, Alexandre, et al. “Genetic Analysis of the beta Subunit of the Epithelial Na+ Channel in Essential Hypertension.” Hypertension, vol.32 No.1, 1998: 129-137.

[vii] Schild, L, E Schneeberger, and I. Gautschi. “Identification of amino acid residues in the alpha, beta and gamma subunits of the epithelial sodium channel (ENaC) involved in amiloride block and ion permeation.” J Gen Physiol, Vol.109, 1997: 15-26.

[viii] Schild, L, E Schneeberger, and I. Gautschi. “Identification of amino acid residues in the alpha, beta and gamma subunits of the epithelial sodium channel (ENaC) involved in amiloride block and ion permeation.” J Gen Physiol, Vol.109, 1997: 15-26.

[ix] Hansson, JH, et al. “Hypertension caused by a truncated epithelial sodium channel gamma subunit; genetic heterogeneity of Liddle syndrome.” Nat Genet, Vol.11, 1995: 76.

[x] Hummler, Edith. “Epithelial Sodium Channel, Salt Intake, and Hypertension.” Current Hypertension Reports, Vol. 5, 2003: 11–18.

[xi] Dong, YB, HD Zhu, and EH Baker. “T594M and G442V polymorphisms of the sodium channel beta subunit and hypertension in a black population.” J Hum Hypertens, Vol.15, 2001: 425–430.

Get Professional Assignment Help Cheaply

Buy Custom Essay

Are you busy and do not have time to handle your assignment? Are you scared that your paper will not make the grade? Do you have responsibilities that may hinder you from turning in your assignment on time? Are you tired and can barely handle your assignment? Are your grades inconsistent?

Whichever your reason is, it is valid! You can get professional academic help from our service at affordable rates. We have a team of professional academic writers who can handle all your assignments.

Why Choose Our Academic Writing Service?

  • Plagiarism free papers
  • Timely delivery
  • Any deadline
  • Skilled, Experienced Native English Writers
  • Subject-relevant academic writer
  • Adherence to paper instructions
  • Ability to tackle bulk assignments
  • Reasonable prices
  • 24/7 Customer Support
  • Get superb grades consistently

Online Academic Help With Different Subjects


Students barely have time to read. We got you! Have your literature essay or book review written without having the hassle of reading the book. You can get your literature paper custom-written for you by our literature specialists.


Do you struggle with finance? No need to torture yourself if finance is not your cup of tea. You can order your finance paper from our academic writing service and get 100% original work from competent finance experts.

Computer science

Computer science is a tough subject. Fortunately, our computer science experts are up to the match. No need to stress and have sleepless nights. Our academic writers will tackle all your computer science assignments and deliver them on time. Let us handle all your python, java, ruby, JavaScript, php , C+ assignments!


While psychology may be an interesting subject, you may lack sufficient time to handle your assignments. Don’t despair; by using our academic writing service, you can be assured of perfect grades. Moreover, your grades will be consistent.


Engineering is quite a demanding subject. Students face a lot of pressure and barely have enough time to do what they love to do. Our academic writing service got you covered! Our engineering specialists follow the paper instructions and ensure timely delivery of the paper.


In the nursing course, you may have difficulties with literature reviews, annotated bibliographies, critical essays, and other assignments. Our nursing assignment writers will offer you professional nursing paper help at low prices.


Truth be told, sociology papers can be quite exhausting. Our academic writing service relieves you of fatigue, pressure, and stress. You can relax and have peace of mind as our academic writers handle your sociology assignment.


We take pride in having some of the best business writers in the industry. Our business writers have a lot of experience in the field. They are reliable, and you can be assured of a high-grade paper. They are able to handle business papers of any subject, length, deadline, and difficulty!


We boast of having some of the most experienced statistics experts in the industry. Our statistics experts have diverse skills, expertise, and knowledge to handle any kind of assignment. They have access to all kinds of software to get your assignment done.


Writing a law essay may prove to be an insurmountable obstacle, especially when you need to know the peculiarities of the legislative framework. Take advantage of our top-notch law specialists and get superb grades and 100% satisfaction.

What discipline/subjects do you deal in?

We have highlighted some of the most popular subjects we handle above. Those are just a tip of the iceberg. We deal in all academic disciplines since our writers are as diverse. They have been drawn from across all disciplines, and orders are assigned to those writers believed to be the best in the field. In a nutshell, there is no task we cannot handle; all you need to do is place your order with us. As long as your instructions are clear, just trust we shall deliver irrespective of the discipline.

Are your writers competent enough to handle my paper?

Our essay writers are graduates with bachelor's, masters, Ph.D., and doctorate degrees in various subjects. The minimum requirement to be an essay writer with our essay writing service is to have a college degree. All our academic writers have a minimum of two years of academic writing. We have a stringent recruitment process to ensure that we get only the most competent essay writers in the industry. We also ensure that the writers are handsomely compensated for their value. The majority of our writers are native English speakers. As such, the fluency of language and grammar is impeccable.

What if I don’t like the paper?

There is a very low likelihood that you won’t like the paper.

Reasons being:

  • When assigning your order, we match the paper’s discipline with the writer’s field/specialization. Since all our writers are graduates, we match the paper’s subject with the field the writer studied. For instance, if it’s a nursing paper, only a nursing graduate and writer will handle it. Furthermore, all our writers have academic writing experience and top-notch research skills.
  • We have a quality assurance that reviews the paper before it gets to you. As such, we ensure that you get a paper that meets the required standard and will most definitely make the grade.

In the event that you don’t like your paper:

  • The writer will revise the paper up to your pleasing. You have unlimited revisions. You simply need to highlight what specifically you don’t like about the paper, and the writer will make the amendments. The paper will be revised until you are satisfied. Revisions are free of charge
  • We will have a different writer write the paper from scratch.
  • Last resort, if the above does not work, we will refund your money.

Will the professor find out I didn’t write the paper myself?

Not at all. All papers are written from scratch. There is no way your tutor or instructor will realize that you did not write the paper yourself. In fact, we recommend using our assignment help services for consistent results.

What if the paper is plagiarized?

We check all papers for plagiarism before we submit them. We use powerful plagiarism checking software such as SafeAssign, LopesWrite, and Turnitin. We also upload the plagiarism report so that you can review it. We understand that plagiarism is academic suicide. We would not take the risk of submitting plagiarized work and jeopardize your academic journey. Furthermore, we do not sell or use prewritten papers, and each paper is written from scratch.

When will I get my paper?

You determine when you get the paper by setting the deadline when placing the order. All papers are delivered within the deadline. We are well aware that we operate in a time-sensitive industry. As such, we have laid out strategies to ensure that the client receives the paper on time and they never miss the deadline. We understand that papers that are submitted late have some points deducted. We do not want you to miss any points due to late submission. We work on beating deadlines by huge margins in order to ensure that you have ample time to review the paper before you submit it.

Will anyone find out that I used your services?

We have a privacy and confidentiality policy that guides our work. We NEVER share any customer information with third parties. Noone will ever know that you used our assignment help services. It’s only between you and us. We are bound by our policies to protect the customer’s identity and information. All your information, such as your names, phone number, email, order information, and so on, are protected. We have robust security systems that ensure that your data is protected. Hacking our systems is close to impossible, and it has never happened.

How our Assignment  Help Service Works

1.      Place an order

You fill all the paper instructions in the order form. Make sure you include all the helpful materials so that our academic writers can deliver the perfect paper. It will also help to eliminate unnecessary revisions.

2.      Pay for the order

Proceed to pay for the paper so that it can be assigned to one of our expert academic writers. The paper subject is matched with the writer’s area of specialization.

3.      Track the progress

You communicate with the writer and know about the progress of the paper. The client can ask the writer for drafts of the paper. The client can upload extra material and include additional instructions from the lecturer. Receive a paper.

4.      Download the paper

The paper is sent to your email and uploaded to your personal account. You also get a plagiarism report attached to your paper.

smile and order essaysmile and order essay PLACE THIS ORDER OR A SIMILAR ORDER WITH US TODAY AND GET A PERFECT SCORE!!!

order custom essay paper