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(NAME ) SADIQ ULLAH (CMS)=ROLLNO 2479 (ASSIGNMENT)=NO 1 (SUBMITTED TO SIR MUBEEN MUSTAFA KIANi)

Basal metabolic rate Metabolic rate Amount of energy used

given period of time


Kcal / h or

in the body in the

Kcal / day

Energy expenditure per unit time

Cal / h Calorie Amount of heat required to raise temperature of 1g of H2O to 1 C 1000 calories = 1 kilo calorie carbohydrates, 4 Kcal / G Ethanol 7 Kcal / G Efficiency of MR 50% ATP proteins, 4Kcal / G fats 9KCal / G

50% Heat

BMR Basal Metabolic Rate it is an estimate of the rate

of metabolism determined by measuring thevolume of respiratory gases generated during a period of time. Energy to support basic body function Energy just to stay alive Minimum cost of existence

The amount of energy expenditure at rest in normal temperature environment in the post absorptive state Post Absorptive State Fasted state Digestive sys is inactive 12h of fasting ? Digestion takes energy Release of energy in this state is sufficient for vital organs Catabolic process.

Absorptive state / Fed state / Anabolism Food fuel energy production & storage

The amount of energy expenditure at rest in normal temperature environment in the post absorptive state Post Absorptive State Fasted state Digestive sys is inactive 12h of fasting ? Digestion takes energy Release of energy in this state is sufficient for vital organs Catabolic process.

Absorptive state / Fed state / Anabolism Food fuel energy production & storage

Human Men

BMR = 1 cal / Kg h 70 kg male , 24 h / day = 1 cal / 70 kg 24 h = 1680 cal / day

: 1 cal / kg / h Women : 0.9 cal / kg /

Factors effecting BMR: Sex BMR then in all ages Age BMR age Height Tall BMR Body Mass Muscle mess BMR Growth Children & pregnant women BMR Body temperature Fever can BMR up to 15% / C Hormones Thyroid hormones BMR

Measuring Exact

BMR:

BMR measurements are difficult. Under restrictive circumstances Person is awake, at complete rest Under less strict condition RMR RMR = Resting metabolic rate i) Direct Calorimetery

Measurment of total heat produced by the body Measure heat = energy Calories / min Living subjects in sealed enclosure ii) Indirect Calorimetery Measurement of the respiratory gases produced by the body Measurement of O2 consumption ( ml O2 / min) Measurement of CO2 production ( ml CO2 / min) Respirometery Drawbacks i) Internal energy ? food fuels ? ii) O2 consumption = CO2 production ?

Respiratory Quotient : (RQ) (R.Q) Complete oxidation of food stuff It is the ratio of CO2 produced to O2 used

RQ = Moles (or ml) CO2 produced Moles (or ml) O2 consumed Carbohydrates: C6H12O6 + 6O2 6CO2 + 6H2O RQ of glucose = 6/6 = 1

/ O2 ) by the body.

(CO2

Fats: C16H32O2 + 23O2 16CO2 + 16H2O RQ of Palmitic acid = 16/23 = 0.696 Proteins: C72H112N2O22S + 77O2 63CO2 + RQ RQ

+ SO3 + 9CO(NH2)2

38H2O

of albumin = 63/77 = 0.818 of Palmitic acid = 16/23 = 0.696

Proteins: RQ ==

C72H112N2O22S + 77O2 63CO2 + 38H2O + SO3 + 9CO(NH2)2 of albumin = 63/77 = 0.818

RQ (arm) (leg) = 0.7 , brain = 1 entire body = 0.82

Brain & RBCs used CHO Little energy from protein, mostly by fats & CHO. Cell memb 1 Cell Membrane Cell Basic unit of St.& function Prokaryotes & eukaryotes Components of cell : Cell membrane Cytoplasm organelles Nucleus

Cell Membrane: The cellis enveloped by a thin membrane Plasma membrane 75 90 thick ECF & ICF Structure & Chemical Composition: Lipids , Protein

Carbohydrates Protein : Lipid 1 : 4 4 : 1

Chemical
Membrane Erythrocytes Out. orembane

Composition of Cell membrane


Proteins% 49 50 75 20 Lipids% 43 46 23 75 Carbohydrates% 8 4 2 5

Inn.Mito membrane Myelin

Fluid Mosaic Model: 1972 S.J.Singer Phospholipid bilayer

fully embedded Freez fracture techniques. Fluid individual phospholipids & protein movement Mosaic pattern produced by proteins Phospholipids
Hydrophilic Hydrophobic

, Nickolson proteins partially or

P FA

heads / Polar tails / non polar

Membrane

Lipids: All are amphipathic molecules


Glycolipids Cholesterol
Major

classes

Phospholipids

Classes

of Phospholipids

Phosphoglycerols

/ Phosphoglycerides Ghycerol backbone Serine, ethanolamine, choline, glycerol ii) Sphingomyelins Sphingosine backbone Myelin sheaths

Lateral movements of phospholipids ?

Glycolipids:
Contains sphingosine Surface, joined by phospholipids head Cholesterol:

Mammalian cells Stability CELL MEMBRANE PROTEINS i) Peripheral proteins ii) Integral Poroteins Peripheral proteins:

extrinsic proteins 30 % attached to the surface by H bonding function as Enzymes & receptors

Integral intrinsic

proteins:

proteins / Transmembrane

proteins Partially or totally immersed in lip bilayer. Passes from 1 side of lip bilayer to other. Some are partly embedded Channels or pores carrier proteins Active transport

CELL MEMBRANE CARBOHYDRATES: combination of CHO proteins & lipid protrude to the outside of the cell the entire outer surface of the cell which has a

loose CHO layer is glycocalaxy Serve as source of connection to other cell. Receptor Functions in antibody processing.

Functions Protective

of Cell Membranes:

function Shape & size of cell Semipermeability MDR : Multi drug resistance

Pumps

MDR proteins = p glycoproteins Cell shows resistance to toxic drugs Shows sensitivity to other drugs MDR

the drug out of the cell

CELL FRACTIONATION The technique used for the: Separation of diff types Subsequent isolation of diff cell organelles Isolate specific cell organelle by the process of centrifugation. i) Preparation of Homogenate Cell lysis or cell disruption Mortars & pestles, blenders, compressors ultrasonificators,homogenizer machines Isotonic solution of 0.25 M sucrose Not metabolized in most tissues Does not cause inter organelle swelling Removal of Debris ii) Differential Centrifugation. The separation of cell organelles by a series of centrifugation steps of time & gravitational force.

Tissue

Homogenete In 0.25 sucrose

Centrifugation

600 g / 10 min

Pellet (Sediment Nuclei)

Supermatant

Centrifugation

15,000 g / 15 min
Pellet Mito, lysosomes, peroxisomes 1, 00,000 g / 60 min

Supermatant

Centrifugation Supermatant Pellet Microsomes

SUBCELLULAR FRACTIONATION OF CELL BY CENTRIFUGATION

Pallet Suppernatant

ISOPYKNIC CENTRIFUGATION TECHNIQUES: density gradient is set up in centrifuge tube. i.e d of sol from top to bottom of tube sucrose percoll sedimentation according to d separation on the basis of d, size & shape MARKER ENZYME: Sucrose
Specific g

sol

PM, ER 1.16

Mito
1.19 Lyso 1.21 Peroxisomes 1.23 Soluble pro

1.30
NA 1.6

to check the purity confirms identify of isolated fraction degree of contamination


Plasma Membrane Nucleus Endoplasmic Teti Golgi Bodies Lysosomes Mitochondria 5 Nucleotidase Na, K ATPase DNA polymerase RNA polymerase G6P Cyto b5 reductase Galactosyl transferase Manosidase Acid phosphatase B Glucuroxidase Succ dehydrogenease Cyto C oxidase Oligomycin Catalase Lact dehydrogenase G 6 Phosphate dehydrogenase

Peroxisomes Cytosol

DIFFERENTIAL CENTRIFUGATION BMB 3.4.1 Based on the differences in the sedimentation rate of the biological particles of different size, shape and density

D glucose & M.p 146 C specific rotation + 112 M.p 150 C specific rotation + 18.7 - 18.7 + 52.7 C 112 Ring Formation:5 (i), 5 (ii), (5iii), (5iv) (5v) Chirality b aCc d

The entire tube is filled with sample and centrifuged Through centrifugation, one obtains a separation of two particles but any particle in the mixture may end up in the supernatant or in the pellet or it may be distributed in both fractions, depending upon its size, shape, density, and conditions of centrifugation Repeat sedimentation at different speed

Medium: same density The sedimentation speed is determined mainly on the size, shape of particle. Application: low resolution separation such as preparation of nucleus

Medium: same density The sedimentation speed is determined mainly on the size, shape of particle. Application: low resolution separation such as preparation of nucleus

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