HUMAN _DATE: ult joa ANATOMY AND Puysy (Beioite) » seG¥ Semester 1 EXPERIMENT NO.:-8 ;- To determine bleeding ti e eding time of own Ai blood sample. she time elapsed between the moment blood escapes from cessation ofits flow is defined as the “bleeding time Bact acd tie jg 1-3 minutes. ‘The usual bleeding time Requirements: piece of filter paper, pricking needle, 70% alcohol. procedure: 1. The tip of ring finger is sterilized with 70% alcohol and a bold prick is given ‘0 that blood flows freely. The stopwatch is immediately started he blood is soaked on a filter paper (While soaking, the filter should not touch the skin of finger) this is repeated every 10 seconds till no blot ‘appears on the papers. ‘The time from the first appearance of the blood to the cessation of bleeding js the bleeding time Discussion: ‘he time clapsed between the moment blood escapes from vessel and the cessation of its flow is defined as the “bleeding time”. The usual bleeding time is 1-3 minutes. The bleeding time is affected by various factors such as the size and shape ofthe injury, endothelial smoothness, the muscularity and elasticity of blood vessels, sealing by platelets, pressure difference between inside and ouside of blood vessel. Blecding time is prolonged during thrombocytopenia, (@eerease in platelet count) and scurvy (Vit, C deficiency) Betas contractility anc platelets play an important role in cesehon bleeding, These are relatively simpler mechanisms clotting, Hence bleeding time is shorter than clotting Gms Clotting of blood and the cessation of bleeding after injury are some of the important haemostatic mechanisms {0 check hemorrhage. These latifbahees may lead to various dieeascs such as heroph than the mechanism of lia, purpura ete. 2022-93 | sNMPPC/ B. Pharmmye HUMAN ANATOMY AND pury; uhijes th} (BPLOITE) LOGY Semester 1 EXPERIMENT No.:.9 gum -To determine clotting or coagulation tinie of own blood between the moment of cua ase ent of escape of blood outside the vessel and th emiopment of fibrin is defined as the “clotting time”, The norm ; ne is 4-30 miNUtCs. requirements: capillaries oF small test tubes or slide, 70% alcohol, cotton, pricking needle se syringe with needle, stop watch principle: During coagulation sol form of the blood is changed to gét from. The time clotting, lapsed between the moment of escape of blood outside the vessel and the gpservation of physical change is taken as clotting time. Procedure: Lee and White’s Method: About 4ce of blood is obtained through a venepuncture and 1 ce. cach of four small test tubes. As soon as blood enters the barrel of syringe, placed in stopwatch is started. The test tubes are placed in water bath at 37 ° C. The test tubes are tilted at every 30 sec. The time when blood does not move on tilting is being noted down. ‘ Wright's Method: ‘The ring finger is sterilized and pricked boldly with the needle. As soon as the blood comes out, the stopwatch is started and a capillary tube is filled upto three-fourth of its total length. After every 30 sec. a portion of capillary tube laboit 1/10 of the tube) is broken until a thin line of unbroken coagultum is Beisteiched between the two broken ends. This time is noted and the diference between the time when blood comes out from t Unbroken coagulum seen, is the coagulation time Duke's Method: The ring finger is sterilized and a drop of dlameter is placed on a special glass slide. The sh 806 The time is noted when no change in the This gives the clotting time. the finger and the blood approximately 5 mm ide is held vertical at every shape of the drop is seen. /snmppcy B, Pharm.: HUMAN ANAToMy ‘ANY . SIOLO gesult and Conclusion; (8P101TP) sem oe Somme OWN blood samp, — mple was fond to be jscussion: zea Ghotting Time: The time between the moment of escape of bloo weseel and the development of fibrin is defined as the “elenng amen ne normal clotting time is 4-10 minutes, Salah The clotting time is shortened by various, factors such as warmth, intense surface agitation and tissue factors. Clotting time is prolonged under various conditions as absence of clotting factor, decrease in temperature, smoothness of surface, dilution of blood ete, Haemophilia is the condition caused by the absence of any one of the factor required for clotting of blood. It causes prolongation of the clotting time. Mechanism of Coagulation (Clotting) ‘Mechanism of coagulation (clotting) The clotting of blood involves following three steps: 1) Formation of thromboplastin: When the blood comes in contact with rough surface or tissue extracts, platelets disintegrate and thromboplastin is generated. 2| Formation of thrombin: Prothrombin (a globulin) and thrombin is formed. Bes of clot: Thrombin thus formed converts fibrinogen to Sbrin ‘The thromboplastin interacts with plasma (oy, but there are no Although the mechanism of clotting appears to be simple, pee : 7 \rombin. BI cg for above reactions except that of the Tormarian © ‘independently by , te Vatious factors involved in the clotting have been ieolg “atious workers.HUMAN ANATOMY AND we ; PHY! Lana ery SIOLOGY 1 niversally. Name of Factor james of the factors accepted Fibrinogen — Prothrombin Thromboplastin | Calcium Accelerator globulin Activated accelerator globulin Stable Factor Anti- hemophilic globulin Christmas factor Stuart's factor Plasma thromboplastin Contact factor i Fibrin stabilizing factor ors except first two are concemed in the which itself is called as the factor-IN. Factor TV (Calcium) is formation of thromboplastin. Calcium is not only essential jon of thrombin and fibrin but it accelerates their formation. .d in various schemes but mong factor V to XIII has been propose rsally acceptable. formation ofHUM: eee i ps ‘AN ANATOMY AND PHYSIOLOGY 1 = (Bpi01n ae }_Semester 1 ‘im: To find out Haemoglobin content and Oxygen carry ‘own blood sample. ata da Requirements: gahli’s haemoglobinometer or hemomet mometer which consists of two sealed comparison tubes fixed in rack, a specially i mparison a k, a specially graduated diluting tube, a thin glass rod (stirer) and micropipette of 20 cubie millimeter (emm) capac pricking needle, absorbent cotton ; a #5 oben OE, asp doe eon ae ee reagents: Reagen “Principal: (Sahli’s method) When blood is mixed with N/10 HCI RBCs are haemolyzed and Hb is liberated. his converted into acid haematin, which is reddish brown in colour. The solution is diluted with distilled water till it matches with the standard glass (comparison) tubes. The Hb% and Hb gm can directly be read from the graduated tube. Procedure: Pears jated diluting tube and the micropipette are cleaned thoroughly and dried the graduated diluting ube is filed with N/10 HC! upto the mark? gm or tithe mieropipette touches the level of acid in the tube. ‘tho ner eleaned with 70% aleaholand itis picked to obtain a drop of hoo, Firat drop ie wiped out. Second drop is sucked inthe micropipette up to the mark 20 emm. oe blood lainey deposited atthe Beto of se grated “ube ‘the pipette is rinsed two to three times in HCL The blood is mixed with the help of stirrer and then solution is allowed to Bre cr vO es nophasl yi crore Wo ee IN? ee pure inn dite it led ne, DAS! WO added ree roy arop ane every ime tie ative ithe xt Bath with standard glass tubes is obtained. , is complete, stirrer is taken out form the dil ibe , When the matchint sand the scale is read on the side of tube: 2022-23 | SXMPPC/ B. Pharm.HUMAN ANATOMY AND PHYSIOLOGY 1 IPLOLTP) Semester 1 DATE: opservations and Calculations: ppserved Hb gr Sea Observed Hb% - -10 International value of Hb is 14.5 gm% = 100% tycatouated wove 225 100 As Yive Tus = 103-44 2) Oxygen Carrying Capacity ‘Amount of O2 in cc. carried by 100 ml of blood during one pulmonary th—> L- circulation (Hb gm*% *1.34 ec) ts 1 gm of Hb contains 1.34 cc oxygen, Therefore, 1C- ~ gm of Hb contains = _LC_* 1.34 = Qo.) ce ones 2022-23 / SNMPPC/ B. PharmHUMAN : Geena YARD PHYSIOLOGy 1 eee ae 101 TP) Semester 1 ut ‘im: - To determine Blood Group of own blood sample, Microscopic slides, glass-marking pencil or pen, Anti-sera A, Ant anti-sera D (Rh), pricking needle, 70% alcohol, microscope. ‘Sera B and Principle: According to ‘ABO! system of blood grouping there can be four blood groups 4,B, AB, and O. This classification is based on the presence of agglutinogens A, B, both (A and B) or none of them respectively. As Landsteiner stated, if particular type of agglutinogen is present in the blood, then the corresponding agglutinin is always absent and if particular type of agglutinogen is absent in the blood, then the corresponding agglutinin is always present. The anti-sera A consists of agglutinin alpha and it causes clumping of RBCs of blood containing agglutinogens A. The anti-sera B consists of agglutinin beta and it causes clumping of RBCs of blood containing agglutinogens B. Hence blood group can be determined as follows. Clumping in anti-sera A Nr Blood group A ‘Clumping in anti-sera B Blood group B Clumping in both anti-sera A&B Blood group AB i No clumping in any of the sera Blood group 0 The blood groups are assigned as +ve or ~ve, according to the 'Rh’ system. If ‘Rh’ factor (a type of agglutinogens) is present ‘the blood group is assigned as +ve and ifitis absent, the blood group is assigned as ~ve. Anti-sera D contains antibodies against ‘Rh factor, hence if clumping is seen in this sera, blood is not seen, it will be -ve group will be +ve an Procedure; 1, Take three slides and mark them A, B and C. 2. Place a drop of ‘Type A ‘serum’ (antiserum A) on slide A and antiserum B on slide B, On the third slide take a drop of anti-D serum. 3. Sterilize the finger and prick it boldly. Place one drop of blood on each of the sera, 4, Mix the cells and the serum with the help of another slide. (Use separate slides or separates edges for mixing each sera) 2022-23 / SNMPPC/ B. Pharm, Page.39 a ll tl lilI wd 8 @ (Be101TP) $ Allow it to react for 5 minutes 6 Examine after S minutes but not later than 10 minutes, The Rot be complete before 5 minutes and drying may occur after 10 Both these factors thay yield false results. observation: Slideno —*Y| Anntti-serum iy aa owe aoe Clumping x Antiserum A RG aT S B Antiserum B US ae c Anti-Rh factor RRS Result: Ce te My blood group is__Q Discussion: When more then 40% of blood is lost over a short period by hemorrhage or so, it has to be restored by intravenous infusion. This is known as blood transfusion. It is an ideal transfusion, as it not only, restores blood volume but also erythrocytes, which may survive for 120 days. However great care must be exercised while selecting person who donates blood (Donor). When an incompatible blood is transfused intravenously from donor to a person who gets blood (Recipient), antibodies (agglutinins) present in recipient's blood bring about agglutination (clumping erythrocytes) due to the antigens (agglutinates) present in donor's blood. This results in severe effects such as haemolysis of erythrocytes, damage of urinary passages, blockage of blood capillaries and death ayHUMAN ANATOMY AND TE PHYSIo at SYSTEM: (BPLOITP) - Semone! Can be pared Pisnaness Agglutinin | Can be ] i ie eee sabe to ee eer |e BE EMME Alpha ——|— al 7 =a one Alphaée Beta | —~ te 7 AZ STE ay Blood group ‘0”is known as Universal donor bese agglutinogen and hence can be donated to any group, ee? AB 12 known as Universal recipient because is doce not contain any agglutinin and hence can receive blood of any group. Now days the terms Universal recipient and universal donor are misnomer. Before transfusion of blood to any patient, cross matching must be done. ABO system follows multiple allele system and hence transferred to eonsecutive generations, according to laws of Mendel. This is used for paternity test in medico legal cases. Rh factor: Apart from agglutinogens A and B, another agglutinogen was discovered in human blood called ‘Rh factor’ or ‘Rhesus factor’, The blood having this factor is described as Rh+ve while the blood without this factor is described as Rh- ve. Almost 90% of world population is Rh+ve. The characteristic of Rh is that agglutinin against Rh antigen is never present in the blood but, is developed after the first exposure to this factor. Thus if Rh+ve blood is given to Rh-ve patient for the first time, anti-Rh antibodies will developing the patient but incompatibility will not take place. However, if second time transfusion is done to the same patient, agglutination may take place. In case of Rh-ve mother bearing Rh+ve foetus, the Rh antigens will travel to mother’s blood from the foetus, As a result Rh+ve antibodies will develop in mother’ blood. Same antibodies may travel back to foetus and antigen- foetus resulting in antibody reaction (Haemolysis) will take place miscarriage (abortion) or if born alive, child suffers from severe anemia (erythroblastosis foetalis). Under these conditions the mother may become sensitized to Rh+ve blood, and agglutination may occur. @ ae 20123 / sumer 2, ram ‘ age 41 |