RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

BANGALORE, KARNATAKA

ANNEXURE –II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR

DISSERTATION

1 / Name of the Candidate
And address
(In block letters)
Permanent Address: / : / DR. SAURABH GUJRATHI
DEPARTMENT OF PATHOLOGY,
NAVODAYA MEDICAL COLLEGE,
RAICHUR – 584103.
DR.SAURABH GUJRATHI
FLAT NO 19, 5 FLOOR, MAHAVIR RESIDENCY, SALISBURY PARK, PUNE-37
2 / Name of the Institution / : / NAVODAYA EDUCATION TRUST’S,
NAVODAYA MEDICAL COLLEGE AND RESEARCH CENTRE,
RAICHUR-584103.
3 / Course of study and subject / : / M.D. PATHOLOGY
(3 ACADEMIC YEARS)
4 / Date of admission to the course / : / 30th MAY 2013
5 / Title of the Topic / : / EFFECTS OF STORAGE ON QUALITY OF PLATELET CONCENTRATES: AN IN VITRO STUDY.
6 / BRIEF RESUME OF THE INTENDED WORK
6.1 / Need of the study :
Human platelets are anucleated cells derived from megakaryocytes and typically circulate for 10 days.1
Platelets are small and discoid in shape, with dimensions of approximately 2.0–4.0 by 0.5µm1, and a mean volume of 7–11 fl. They are the second most numerous corpuscles in the blood, numbering between 150–450 x 109/L.1
Platelets are surprisingly multifunctional and are involved in many pathophysiological processes including haemostasis, thrombosis, clot retraction, vessel constriction and repair, inflammation including promotion of atherosclerosis, host defence and even tumour growth/ metastasis.1
There is increased demand for stored platelet concentrates (PCs) for therapeutic transfusions such as treatment of patients with disorders resulting in thrombocytopenia and for patients who become thrombocytopenic after chemotherapy or major invasive procedures, such as cardiac surgery. High platelet quality would be expected to result in improved clinical efficacy, determined by count increment, improved hemostasis, and lower risk for adverse reactions in recipients.2
Loss of the platelet functionality due to platelet activation occurs during preparation and storage of platelet concentrates referred as “platelet storage lesion” (PSL).2 Such changes are associated with decreased hemostatic function of platelets after transfusion and with poor post-transfusion recovery. The platelet activation has been demonstrated by changes in platelet morphology, biochemistry, granule release, and surface glycoprotein expression. Hence, the quality of platelet concentrates (PC) is an important issue in transfusion therapy.2
Platelets are stored at 22-24° C for a maximum of 5-7 days.3 There are changes in platelet parameters like pH, Mean platelet volume, Platelet count, Lactate Dehydrogenase levels, Glucose levels and bacterial contamination of platelet concentrates during storage.4,5
Hence, this study is aimed at exploring and documenting the change in quality of platelet concentrates due to storage at the Blood Bank Navodaya Medical College and Research Centre, Raichur.
6.2 / Review of Literature:
Platelets were first described by the remarkably early observations of Bizzozero.1 Dr. Scott Murphy and Frank Gardner provided evidence that platelets could be stored at 22 ± 2°C, for up to 3 days.6 Wright was the first to identify the megakaryocyte as the precursor cell to the platelets.7
Moroff et al. suggested that a specific platelet subpopulation comprising a small portion of all the platelets in platelet concentrates prepared from platelet-rich plasma was responsible for most of the drop in pH.4
Skripchenko et al. concluded that prolonged periods of elevated carbon dioxide levels are associated with poor pH control and platelet mitochondrial dysfunction during storage.4
Platelets are separated from whole blood from donors with standard donation criteria with written informed consent according to institutional guidelines. A total of 350 or 450 ml of whole blood is drawn into triple or quadruple blood bag. Within 2-6 hours of collection, the whole blood units are centrifuged in refrigerator centrifuge at 1500 rpm for 7 minutes. After centrifugation, platelet rich plasma transferred to satellite bag. The platelet rich plasma is centrifuged again at 3000 rpm for 10 minutes to get platelet concentrate.8
The platelet counts were normal throughout storage period. No difference in platelet counts detected during storage. Differences in mean platelet volume were detected on days 3, 5 and 7 of storage. The extracellular Lactate Dehydrogenase activity remained stable throughout storage period.4 From day 3 of storage of platelet concentrates, levels of pH and glucose concentration were lower.4
Swirling movements remained in platelet concentrates at the highest level for 5 days in all reference units throughout the storage period. After the fifth day of storage, all the investigated groups of platelet concentrates lost swirling movements.4
Bacterial contamination was seen in few platelet concentrate samples. There is risk for bacterial contamination, because the standard storage conditions for platelets consist of incubation at 22oC, most commonly suspended in plasma, ideal conditions for the growth of most bacterial species. Thus, the inoculation of only a few organisms by inadequate skin preparation during donation can lead over the time of storage to the production of large numbers of bacteria.9
Pathogenic bacteria likeStaphylococcus aureus, coagulase negative Staphylococci,viridans group Streptococci,Bacillus spp.,Corynebacteriaas well as anaerobic diphteroid gram positive bacilli such asPropionibacterium acnes were detected in platelet concentrates during storage. 10
6.3 / Objectives of the study:
To study in vitro changes, associated with quality of platelet concentrates on storage before transfusion.
7 / MATERIALS AND METHODS:
7.1 / Source of Data:
All the platelet concentrates prepared in Blood bank of Navodaya Medical College, Hospital and Research Centre, Raichur.
7.2 / Methods of collection of Data (including sampling procedure, if any)
Study design: Prospective In vitro study.
Study period: October 2013 to September 2015.
Sample size: 100 samples.
Method. After preparation of platelet concentrates by centrifugation, a sample of well mixed platelet concentrates will be examined every alternate days from day 0 to day 9. The parameters like: pH, Platelet count, Mean platelet volume, Lactate Dehydrogenase levels, Glucose levels, Swirling movements, Bacterial culture will be recorded as per the standard procedures. The collected data will be analysed by statistical analysis by calculating mean, standard deviation. Statistical significance will be calculated by student “t” test.
Inclusion criteria:
·  Randomly selected platelet concentrates prepared in blood bank of Navodaya Medical College, Hospital and Research Centre, Raichur.
Exclusion criteria:
·  Platelet concentrates contaminated with red blood cells and low volume concentrates.
7.3 / Does the study require any investigation or intervention to be conducted on patients or other humans or animals? If so, please describe briefly.
No.
7.4 / Has ethical clearance been obtained from your institution in case of 7.3?
Yes.
8 / LIST OF REFERENCES :
1)  Harrison P. Platelet function analysis. Blood Reviews 2005; 19:111- 123.
2)  Hegazi M, Qari M. The influence of exposure to light and dark environment on platelet concentrates during storage. Alexandria bulletin 2007; 43:715-16.
3)  Gitz E et al. Improved platelet survival after cold storage by prevention of glycoprotein clustering in lipid rafts. Hematologica 2012; 97 (12): 1873- 81.
4)  Gulliksson H, Sandgren P, Sjodin A, Hultenby K. Storage of platelets: effects associated with high platelet content in platelet storage containers. Blood Transfus 2012; 10: 205 – 12.
5)  Neiva T J C et al. Evaluation of platelet aggregation in platelet concentrates: storage implications. Rev. bras.Hematol 2003; 25 (4): 207-12.
6)  Blajchman M A. Platelet transfusions: An historical perspective. Hematology 2008:197.
7)  Coller B S. Historical perspective and future directions in platelet research. J Thromb Haemost 2011; 1- 38.
8)  Slichter S J, Bolgiano D, Corson J, Jones M K, Christoffel T. Extended storage of platelet rich plasma prepared platelet concentrates in plasma. Transfusion 2010; 50 (10): 1- 22.
9)  Serrano K, Devine D V. The platelet storage lesion. Clin.lab med 2010 ;( 30): 477-87.
10) Vedy D, Robert D, Gasparini D, Canellini G, Waldvogel S, Tissot JD. Bacterial contamination of platelet concentrates: pathogen detection and inactivation methods. Hematol Rev.2009; 1(1): 5.
9. / Signature of the Candidate
10. / Remarks of the Guide / This study emphasis on quality of platelets concentrates during storage period in vitro, and may help modify in vitro environment for better quality of platelet concentrates.
11. / 11.1 / Name and Designation of
Guide (In block letters) / DR.BASAVARAJ.P.B. M.B.B.S, M. D
ASSOCIATE PROFESSOR.
DEPARTMENT OF PATHOLOGY,
NAVODAYA MEDICAL COLLEGE,
RAICHUR-584103
11.2 / Signature
11.3 / Co-guide (if any)
11.4 / Signature
11.5 / Head of the Department / DR.NARASIMHA MURTHY M.B.B.S, M. D
PROFESSOR AND H.O.D.
DEPARTMENT OF PATHOLOGY,
NAVODAYA MEDICAL COLLEGE,
RAICHUR-584103.
11.6 / Signature
12 / 12.1 / Remarks of Chairman and Principal
12.2 / Signature