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Red Blood Cells and their Function

Blood found in humans as well as other species transports waste products of metabolism aside from the cells while also delivering requisite nutrients around the body as well as oxygen. Blood cells dissolved in plasma or serum make up that very same substance. Plasma is the major medium for excretion product and transportation mostly has been major amount of liquid or water, which is 92% volume as well as encompasses proteins, glucose, essential minerals, hormones, also carbon dioxide, as well as blood cells (Bain, 2022). Plasma in blood encompasses around 55% of the fluid in body. Albumin is the major abundant protein inside the plasma, and the main function is to regulate blood's colloidal osmotic pressure. Red blood cells (also known as RBCs or even erythrocytes), white blood cells (also known as WBCs or  rather leukocytes), as well as platelets (also referred to as thrombocytes) are the common blood cells (Garraud and Tissot, 2018).

Red blood cells are by far the most numerous cells throughout vertebrate blood. Those same encompass haemoglobin, an iron-containing protein that helps carries oxygen whilst also reversibly binding as well as massively increasing the solubilization throughout blood (Pillai et al., 2020). Haemoglobin, an iron-containing oxygen transporting metalloprotein found among almost every vertebrates' red blood cells or erythrocytes and some invertebrates' tissues. RBCs contain haemoglobin, which transports oxygen around the body towards the remainder of the body which is the tissues (NCBI, 2014). It then discharges the oxygen, allowing aerobic energy system or respiration to focus on providing energy towards that same organism's activities and functions during the procedure termed as metabolic activities or metabolism. In each and every 100 millilitres of blood in a normal adult would have 12 - 20 grammes of haemoglobin. In adults of haemoglobin up to four oxygen molecules could be coupled or carried by the mammalian haemoglobin protein. Haemoglobin is also engaged inside the travel of many other particles: it transports concerning 20 to 25 % of bodies natural respiration, carbon dioxide as carbamino haemoglobin, wherein the CO2 is bonded towards the heme protein (Costanzo, 2007). There are multiple haemoglobin genes in humans: HBA1, HBA2, as well as HBB coded for haemoglobin A, the much more common protein in red blood cells found in adults. The genes HBA1 as well as HBA2, which are also on chromosome 16 as well as nearer to one another, codes for the haemoglobin subunits alpha 1 as well as alpha 2 (Uniprot, 2022). The HBB gene found on chromosome 11 codes for the haemoglobin subunit beta. In haemoglobins, the amino acid sequences of globin proteins differ substantially among species. To increasing human evolution path length among species, the above distinctions become much more noticeable. Many multiple subunit of the globular proteins, including haemoglobin, have the quaternary structure. In haemoglobin, the significant proportion of amino acid residues shape alpha helices, which are linked up by shorter form of non-helical sections. The helical segments of all this protein are stabilised by hydrogen bonds, that also induce attractions within in the particle, starting to cause every polypeptide chain towards and fold as well as deform into a specific pattern and shape (Marengo-Rowe, 2006). Haemoglobin’s quaternary structure is made up of four subunits arranged in an approximately tetrahedral pattern. The haemoglobin molecule is organized into four globular polypeptide subunits throughout most vertebrate species. A protein sequence is strongly connected to a non-protein prostheses heme group within every subunit. Inside a globin fold configuration, for every protein sequence is organised into such a series of alpha-helix functional sections that are associated around each other.

The Composition of Plasma and its Role in the Body

A blood type is categorization of blood determined by the presence as well as lack of antibodies but also hereditary antigenic materials on the cell surface of RBCs. Based  upon that blood type system, those same antigens can be proteins, carbohydrates, glycoproteins, or glycolipids (NHS, 2020). Most of the antigens could also be placed on the cell surface of many other cell types in different body tissue. Almost all of the antigens present on the surface of RBCs could come from the same allele or even different version of the very same genotype as well as establish a blood type group system. Two antigens as well as two antibodies reported in blood plasma make up the ABO blood group framework, an Antigen A as well as antigen B are the respective two forms of the antigens. Antibody A as well as the antibody B are the antibodies are the two forms of antigens (FDA, 2021). Antigens are observed on RBCs, as well as antibodies have been found inside the serum. All humans could be divided into four categories based on their blood antigen properties: those who have antigen A are known to be group A), those who have antigen B are classified as group B), those who are both the known antigen A as well as B are categorised in group AB, as well as those with neither the antigen A nor B are grouped in AB , or group O (Xu et al., 2019). Antibodies identified in conjunction within these antigens were: Antigen A present with the antibody B, Antigen B present with the antibody A, Antigen AB with the lack of no antibodies, as well as Antigen nil or group O with the antibody A are all examples of antigens with antibodies. To 50 antigens, the Rh system, with Rh implying Rhesus is the second greatest blood-group structure in living person. The D antigen is by far the most essential Rh antigen which is the more likely to elicit an immune reaction of the present five major Rh antigens (Redcrossblood, 2021). Anti-D IgG as well as IgM antibodies have all been rarely generated by the sensitization towards the environmental stimulants, as such anti-D antibodies have been rarely found in D-negative community members. Even so, a sensitising occasion, such as the fetomaternal transfusion of the blood from that of a foetus or baby present during pregnancy or even during a blood transfusion to D positive RBCs, could indeed cause D-negative individual people towards the production of IgG anti-D antibodies.

Thrombin would be a serine protease molecule which is encrypted by that of the F2 gene found inside the humans. Throughout the clotting of blood process, prothrombin the coagulation factor II, is mostly proteolytically cleaved to produce thrombin (Cleveland Clinic, 2022). Thrombin, throughout turn, functions as just a serine protease, converting solubilized fibrinogen into the insoluble fibrin layers as well as catalysing a variety of many other coagulation-related responses. The ABO blood group system is found on the concept of the agglutination. It would be the most widely used technique for evaluating the appearance as well as utter lack of the cellular antigens as well as their corresponding antibodies inside the blood. Forward typing throughout blood typing refers to the identification of antigen inside the donor's RBCs. Reverse typing, and from the other hand, has been the identification of antibodies inside the donor's serum or plasma (Dean, 2005). The Abo blood system's antigens are glycolipid-based, as well as the antibodies have become mostly of the IgM type.

Understanding Blood Types and Antigens

Type A, B, AB, as well as O phenotypes represent approximately the ABO blood system, that would be used to recognize the nature of exterior antigen - antibody existing inside a participant's blood. Whether the RBCs agglutinate, the commensurate antibody would have to be lacking from the plasma proteins (Ewald, and Sumner, 2016). Only when antigens are not found on the membranes of RBCs, antibodies would have to be available inside the plasma proteins. Agglutination reactions take place seen between the exterior proteins that are involved upon that RBCs exterior inside the ABO blood classi?cation system. The above antigen or agglutinogen would be a surface protein. Antigens also seem to be primarily glycolipids, which have been classified as either self-antigens and otherwise foreign-antigens. Whereas if antigen seems to be distant, the immune response inside the body, as well as the WBCs are activated to recognise these antigens. The WBCs whereupon begin to produce antibodies against such an antigen. The prothrombin time (PT) would be a blood calculation that evaluates how quickly blood clots. To verify for haemorrhagic issues, a PT test could be used. PT can also be used to see if blood clot-prevention medication is effective. An INR test is another name for a PT test., the INR (international normalised ratio) is a technique of standardising PT testing results regardless of measurement technique. For the blood coagulation, clotting components are essential (UCSF, 2019). Prothrombin, also known as factor II, would be a clotting component secreted by the bodies liver. Prothrombin as well as other coagulation factors require vitamin K to be produced. The PT test is very important since it verifies for the presence of five distinct blood coagulation, factors being I, II, V, VII, as well as X). The following factors prolong the PT test: warfarin, for example, is a blood thinner, blood coagulation factors are at an all-time negligible,  the action of some of the coagulation factors keeps changing, some of the coagulation factors are missing, inhibitors are other compounds which actually effect the coagulation factors, as well as any use of coagulation factors has increased. An anomalous PT test is generally triggered by the liver illness or disorder, as well as blood thinner therapeutic interventions.

The following reaction to occur begins with the collection of specimens in the correct tube, the main reason being collection in any wrong tube could alter the pathway and the experimental procedure. As the test requires blood which are not coagulated the tube (light blue) containing EDTA (Ethylenediaminetetraacetic acid) whose main function in these procedures is to bind with the metallic ions for instance iron, magnesium, calcium as well as lead. The EDTA acts as a chelating agent which hinders the formation of blood clot as well as removes the presence of lead from the body. The EDTA tube contains the blood sample, in a slide 5 circles were drawn  and labelled as A,B, D1 and D2 and – (blank). ~50ul of antiserum was added then one drop of blood was added and mixed with yellow tip then observed for the formation of agglutination. The PT test was obtained in a sodium citrate tube and centrifuged at 2000 rpm for 15 minutes. The samples are stored at -20 ºC for longer time, as storage solution for the tests. The thromboplastin D was warmed at temperature of 37 ºC, the tubes were labelled with ID number and 25ul of the plasma was added and warmed for 2 minutes. The reagent measuring 50ul was added to the plasma and the clotting formation was observed. Carefully the reaction was carried to avoid contamination of any other foreign sources to avoid formation of fibrinogen or substances.

The Rh System and Anti-D Antibodies

AQ1. Antibodies for antigens A and B accumulate in plasma and alluded as anti-A and B sera. Some of the antigens can be mixed together producing blood group AB (Palomar, 2014)

AQ2. The appearance of agglutination after the addition of the reactants within the given normal time frame (Mycts, 2022).

AQ3. Assay done at room temperature or lower maintains the reactants, higher than that weakens the reactions.

AQ4. The technique to identify the ABO blood group based on the presence of anti A or anti B sera.

AQ5. Each person either has anti D negative or positive as they vary due Rhd protein; one is developed during in womb, other later.

AQ6. The ABO blood grouping are A, B, AB, and O group, and with Rh divides it into positive and negative types.

BQ1. The PT measures the time and rate at which blood coagulates in sample, and PTT monitors an characteristics of coagulation of blood.

BQ2. Haemoglobin, lipids, bilirubin, and paraproteins, or any condition interferes with PT test.

BQ3. Liver disease, factor VII and vitamin K deficiency, also coagulation factor deficiency could delay coagulation in samples.

BQ4. Heparin present in sample or contamination in samples along with thromboplastin could alter results. Blood when not stored at correct temperature losses their function mainly, coagulation.

BQ5. Due to sample storage at different temperature the prothrombin time gets delayed or increases than usual time compared to other samples.

The ABO blood grouping test for the three samples indicates; S1 is anti A negative thus the presence of antigen A and anti D negative, S2 indicates anti B sera presence and anti D negative sera thus the presence of B- blood group, and for S3 from the test indicated the presence of the anti A and B sera along with anti D sera, which shows the presence of blood group O positive (Table 1). The PT test for the different plasma and blood kept at different temperature indicates the formation of blood and plasma coagulation ; -80 has clotting time of 30 seconds whereas blood and plasma kept at 4º C coagulates at time of 25 sec and 19 sec respectively.

Sample ID

Anti A

Anti B

Anti Rhd D1/D2

-

Blood group

S1

+

-

-

A-

S2

-

+

-

B-

S3

+

+

+

O+

Table 1: ABO blood grouping of sample 1, 2, and 3

Sample ID

PT

INR

Comment

Plasma -80

30 sec

6.07

Visible loss clotting factor

Plasma 4 C

25 sec

4.53

Almost loss of clotting factor

Blood 4 C

19 sec

2.49

Little loss of clotting factor

Table 2: PT test, indicating the presence of blood coagulation time after the addition of thromboplastin D, also delay in PT as normal range is 11 to 13.5 seconds

References

Bain, B.J., 2022. Blood cells: a practical guide. John Wiley & Sons.

Cleveland Clinic, 2022. Prothrombin Time and INR. [online] Available at: <https://my.clevelandclinic.org/health/diagnostics/17691-prothrombin-time-pt-test> [Accessed 3 January 2022].

Costanzo, L., 2007. Physiology : Linda S. Costanzo : Free Download, Borrow, and Streaming : Internet Archive. [online] Internet Archive. Available at: <https://archive.org/details/physiology00cost_0> [Accessed 3 January 2022].

Dean, L., 2005. The ABO blood group. [online] Ncbi.nlm.nih.gov. Available at: <https://www.ncbi.nlm.nih.gov/books/NBK2267/> [Accessed 3 January 2022].

Ewald, D.R. and Sumner, S.C., 2016. Blood type biochemistry and human disease. Wiley Interdisciplinary Reviews: Systems Biology and Medicine, 8(6), pp.517-535.

Garraud, O. and Tissot, J.D., 2018. Blood and blood components: from similarities to differences. Frontiers in medicine, 5, p.84.

Marengo-Rowe, A.J., 2006, July. Structure-function relations of human hemoglobins. In Baylor University Medical Center Proceedings (Vol. 19, No. 3, pp. 239-245). Taylor & Francis.

Mycts, 2022. [online] Mycts.org. Available at: <https://www.mycts.org/Portals/0/Assay_PI/WholeBlood/ABO%20Reagent%20RBC.pdf> [Accessed 3 January 2022].

NCBI, 2014. Haemoglobin and iron: information for blood donors. [online] Ncbi.nlm.nih.gov. Available at: <https://www.ncbi.nlm.nih.gov/books/NBK310577/> [Accessed 3 January 2022].

NHS, 2020. Blood groups. [online] nhs.uk. Available at: <https://www.nhs.uk/conditions/blood-groups/#:~:text=There%20are%204%20main%20blood,there%20are%208%20blood%20groups.> [Accessed 3 January 2022].

Palomar, 2014. Human Blood: ABO Blood Types. [online] Www2.palomar.edu

Pillai, A.S., Chandler, S.A., Liu, Y., Signore, A.V., Cortez-Romero, C.R., Benesch, J.L., Laganowsky, A., Storz, J.F., Hochberg, G.K. and Thornton, J.W., 2020. Origin of complexity in haemoglobin evolution. Nature, 581(7809), pp.480-485.

Redcrossblood, 2021. What Is The Rh Factor? Why Is It Important?. [online] Redcrossblood.org. Available at: <https://www.redcrossblood.org/local-homepage/news/article/what-is-the-rh-factor--why-is-it-important-.html> [Accessed 3 January 2022].

Redcrossblood, 2021. What Is The Rh Factor? Why Is It Important?. [online] Redcrossblood.org. Available at: <https://www.redcrossblood.org/local-homepage/news/article/what-is-the-rh-factor--why-is-it-important-.html> [Accessed 3 January 2022].

UCSF, 2019. Prothrombin Time (PT). [online] ucsfhealth.org. Available at: <https://www.ucsfhealth.org/medical-tests/prothrombin-time-(pt)> [Accessed 3 January 2022].

Uniprot, 2022. HBB - Hemoglobin subunit beta - Pan paniscus (Pygmy chimpanzee) - HBB gene & protein. [online] Uniprot.org. Available at: <https://www.uniprot.org/uniprot/P68872> [Accessed 3 January 2022].

Xu, X., Xu, F., Ying, Y., Hong, X., Liu, Y., Chen, S., He, J., Zhu, F. and Hu, W., 2019. ABO antigen levels on platelets of normal and variant ABO blood group individuals. Platelets, 30(7), pp.854-860.

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