Alopecia Areata: Summary of Presentation - Lab Report Example

Alopecia Areata: Summary of Presentation Introduction Alopecia areata is a medical condition by which the hair in the body is lost especially in thescalp. There are various cycles in which hair growth occurs. One of which is anagen, otherwise known as the growth phase of the hair follicle. The next phase is the telogen or the resting phase. It is during telogen phase where the hair stops to grow. It should be noted that alopecia areata attacks exclusively anagen follicles and oftentimes, lymphocytic infiltrates are observed around and within lower hair follicle. Alopecia areata affects male and female and all races equally. The lifetime risk for acquiring this condition is 1.7%. Alopecia areata is caused by several factors, including microbial, psychological distress, and autoimmune disease. It is associated with HLA class II gene. Pathogenesis of alopecia areata can be confirmed via T – cells (cell – mediated) or autoantibody (humoral) response to hair follicles. Alopecia areata comes in various forms, inclusive of: Diffuse or patchy alopecia areata, alopecia areata totalis, and alopecia areata universalis. An exclamation point hair is the clinical presentation of the hair that is found in alopecia areata. It should be noted that compared with the normal type of hair, the exclamation point hair in alopecia areata lacks the bulb that is found in the normal hair. Objective of the Study (1) To identify the autoantigen of alopecia areata by separating the antigen from whole hair follicle extract, (2) To identify the antigen with proteomics, and (3) to identify the functional relevance of antigen. Sample Used (a) Normal hair follicle extract was from normal plucked hair, extracted with 6M urea (b) Alopecia areata patient serum (n – 10) (c) Normal serum (N – 10) Methods used and the Results A. Method Optimisation (first scheme) (a) Separation of alopecia areata associated antigen from hair follicle extract by 2D SDS – Page. The IgG was purified from whole alopecia areata serum and the purified IgG was then used to immunoprecipitate antigen in the normal hair follicle extract. (b) The hair follicle extract that was separated was visualised using a special stain. Gel one is visualised using Coomassie stain or silver stain. Gel two is visualised using Western Blotting. (c) The isolation of protein spots from gel 2 corresponds to positive spots from western blot for proteomics analysis. Results: Nothing was detected in Western Blotting using commercial anti – human IgG antibody and Western Blotting using purified IgG. Thus, purification step was abandoned because it yields low IgG, and instead whole alopecia areata serum was used for immunoprecipitation. As a result, positive bands of protein were detected in Western Blotting using commercial AE1 and AE3 anti – human keratin antibody and whole alopecia areata serum. Through Peptide Mass Fingerprinting, the following were visualised using the Silver and Coomassie blue stains: Blank (clean), A1 and A2 (Keratin 14), C 1 (Keratin 10), and B2 (K16). B, Method Optimisation (second scheme, using gels and Western Blotting mirror) Identification of the antigen Results: In the second scheme, the following were identified in Western Blotting: (a) Blank – K1 and K10 identified as common contaminants (b) B – K 85 identified as hair – specific keratin (c) C – K 81, K 83, K 86 identified as hair - specific keratin C. Method Optimisation (third scheme, no gels) (a) Separation of alopecia areata associated antigen from hair follicle extract by whole alopecia areata serum with x – linked immunoprecipitation (b) Identification of all antigen in immunoprecipitate using proteomics approach Results: The following were visualised: (a) Trichohyalin (b) Actin and Tubulin (c) Complement C4a (d) HSP 27 (e) Cyclophilin A (f) Calmodulin – like protein 3 More patients and normal samples were recommended using this scheme. Functional Relevance Antigen for alopecia areata can be identified using whole alopecia areata serum. Types of Control Used Positive control: Heat inactivated whole alopecia areata serum Negative control: Heat inactivated fetal calf serum and heat inactivated whole normal serum Conclusion From this study, it was noted that the second scheme was successful and was positive for antigen identification. One can therefore conclude that antigen for alopecia areata can be identified using proteomics approach. Through autoantigen identification, the causative factors for alopecia areata was identified. Any Improvements Needed To be able to find any commercial antibody against the identified antigen of alopecia areata such as the commercial antibody against potential alopecia areata autoantigen trichohyalin incubated with normal hair follicle. Further recommendation to culture hair follicle is presented. In order to accurately identify the timeline of the onset of alopecia areata, it is recommended that an alopecia areata mouse serum must be collected every month since birth until such time that the mouse will first develop the first patch of alopecia areata (should include before, during and after the development of alopecia areata). Reflective Statement: Biomedical science is an applied science that deals with health and is considered a research - oriented field. It involves a variety of medical fields. Its great contribution is in the field of immunology and is successful in the treatment of cancer. For this reason, seminars among postgraduate students are inevitable and oftentimes beneficial. Seminars are also relevant among postgraduate students to inculcate in their minds the importance of biomedical science towards the evolution of disease treatment. To understand better its purpose amongst the students, one must be able to formulate a learning objective of the seminar. As a postgraduate student, I was able to formulate my own learning objective in attending the seminar of biomedical science. The following learning objectives are: (1) To increase my knowledge on biomedical science (2) To update myself on the evolution of medical treatment (3) To be able to witness the scientific way of presentation (4) To be able to learn from the topics being presented (5) To gain knowledge from the presentation itself and use it as my future reference As a biomedical science student, I learned to use the word discipline in every endeavour that I have. I learned how to discipline myself in gaining knowledge through reading books and journals. I also learned to discipline myself on how to manage my time properly especially in the area of research. I was able to realise that time alone is not enough nor discipline to learn and understand the complexity of biomedical science; however, time and disciple must come together in such a way that one cannot exist without the other. Through then, I discovered the importance of good preparation and rehearse. Proper time management and discipline to oneself enables me to prepare for my future research and be able to rehearse on how to present my topic with enormous knowledge and wisdom. As the saying by Benjamin Franklin goes, “By failing to prepare, you are preparing to fail.” A biomedical science student cannot submit a research study in an overnight preparation. Moreover, a biomedical science student cannot present an informative topic with only an overnight preparation for the presentation. Instead, a considerable amount of time is needed to gather the materials necessary for the research, to do the research paper, and to present an informative research paper. A good preparation involves a considerable amount of time and discipline. Preparation involves investment of time and effort. No one fails when he prepares. And of course, proper and constant communication with my research supervisor will complete the ingredients. My supervisor will serve as my mentor, my guide, my conscience, and my critique. No one shares his knowledge and idea to a neophyte like me except my supervisor. It is important that a biomedical science student should allocate his time to see his supervisor and be disciplined enough to listen to his teachings and willingly apply his suggestions. No one is willing enough to lend his ears and shares important information but him. After all, knowledge involves constant quest and that does not happen overnight when you are alone. Read More