BIOCHEMISTRY DEMONSTRATION POWERPOINT PR

BIOCHEM

1. BIOCHEMISTRY

2. DEFINITION BIOCHEMISTRY , ALSO called as BIOLOGICAL CHEMISTRY OR PHYSIOLOGICAL CHEMISTRY is the study of chemical process in the living organism . Also called as biological chemistry or physiological chemistry is the study of chemical process in the living organisms. It can also be defined as the study of molecular basis of life .

3. Biochemistry teaches how the biological molecules like carbohydrates, proteins, lipids, nucleic acids gives rise to different chemical process in the living cell which in turn gives rise to the complexity of life. It was first proposed by a GERMAN scientist, CARL NEUBERG , in 1903. It emerged as a distinct discipline around the beginning of the 20 th century when scientists combined chemistry, physiology, and biology to investigate the chemistry of life.

4. SIGNIFICANCE IN NURSING As the definition says that biochemistry is the study of chemical process in the living cell, it is very essential that a nurse being a medical professional , study and understand biochemistry in order to care for their patients .

5. Knowing the normal process that occur within the body will help us to IDENTIFY THE ALTERATIONS that can occur during disease conditions so that an effective medical treatment can be facilitated. The various biochemical tests like blood test, kidney function test, liver function test, lipid profile, will help the nurse/healthcare provider to diagnose diseases and the test results will help in assessing the needs of the patient to provide an effective care.

6. EXAMPLE: A patient was admitted to the hospital and during the stay in the hospital he suddenly comes up with certain symptoms like confusion, dizziness, shaky feeling, pounding heart, racing pulse, sweating, trembling and weakness. The medical history show that he has diabetes mellitus and was taking insulin. A nurse will be able to understand immediately that the cause of his symptoms were due to hypoglycemia and she will be understand the changes that can occur in the body and will be able to give the timely treatment.

7. HYPOGLYCEMIA is a serious condition in day to day life if not taken care, as is may lead to life threatening complications affecting cardiac and central nervous system. There is also a chance that the patient may go into seizure and coma. So, a thorough understanding of the biochemical processes taking place in the body will help the nurse to have a patient-centered approach in the treatment.

8. Some importance of biochemistry in nursing: Knowing the normal processes that occur within the body will help us to identify the alterations that can occur during disease conditions so that an effective medical treatment can be facilitated.

9. BIOCHEMISTRY teaches about HOMEOSTASIS AND ELECTROLYTE IMBALANCE and a nurse should understand the of it in the body. - All the organs and tissues in the body function to maintain this homeostasis. - Certain disease conditions may create an imbalance of one or more electrolytes which is a serious condition. - The biochemistry teaches about the normal and abnormal metabolite levels and this knowledge helps the nurse to take special care of the patient by maintaining the fluid and electrolytes.

10. HOMEOSTASIS THE ability or tendency of the living organism, cell, or group to keep the condition inside it the same despite any changes in the conditions around it, or this state of internal balance.

11. The various BIOCHEMICAL TEST like blood test, kidney function test, liver function test, lipid profile, will help the nurse to diagnose diseases and the test results will help in assessing the needs of the patient to provide an effective care . Every drug has a biochemical actions and this knowledge will help in SELECTING THE APPROPRIATE DOSES and the duration of treatment a patient requires. NUTRITIONAL BIOCHEMISTRY teaches the relation between health diet and disease. > it focuses on how each nutrient functions metabolically > using biochemical tools nutritional deficiency diseases can be diagnosed. NOTE: BIOCHEMISTRY is important in nursing because it gives knowledge about normal chemical process within the living cells and any change in it helps in the diagnosis of disease, thereby helping the nurse to provide an effective care to the patient.

12. THE CELL ALL ORGANISMS ARE MADE UP OF CELLS “IT IS THE BASIC STRUCTURAL, FUNCTIONAL and BIOLOGICAL UNIT OF LIFE.” IT WAS DISCOVERED BY ROBERT HOOKE IN 1665

13. The word cell comes from a Latin word ‘CELLA’ meaning “ SMALL ROOM”. CELLS are the building BLOCKS of life where all chemical process occurs. They take up the BIOLOGICAL MOLECULES, convert them into energy or use them to carry out specialized FUNCTIONS. It also contains the hereditary material which determine the genetic characteristics

14. The cell theory developed by mathias Jacob and theodor schwann “ All organisms are composed of one or more cells, that all cells come from pre-existing cells, the vital functions of an organism occur within the cells and that all cells contain hereditary information necessary for cell functions and for transmitting from one generation to next.”

15. Every living things (animals, plants, bacteria, fungi protozoans) are made up of cell . Some organisms are made up of just one single cell . ( Unicellular organism e.g BACTERIA). While some organisms are made up of many cells . ( multi cellular organisms e.g. ANIMAL CELL).

16. The human body is built of about one trillion cells specializing in different functions. Many identical cells joins together and forms a TISSUE. Various Tissues , that perform a particular function organize together to form an organ. Various organs joins together to form an organ system .

17. Types of cells EUKARYOTIC CELL 2. PROKARYOTIC CELL

18. EUKARYOTIC CELL Are those cells which have a true NUCLEUS It has a nuclear membrane within which there is well defined chromosomes It has other membrane bound organelles like mitochondria, endoplasmic reticulum, Golgi bodies etc.

19. Organisms with  eukaryotic cells are called as Eukaryotes. They may be single-celled or multicellular organisms. ( e.g.cells of plant, animals, fungi.)

20. PROKARYOTIC CELL •  Prokaryotic cells are those cells whose nucleus is not distinct and their DNA is not organized in to chromosomes. • They lack most of the membrane bound organelles. • The organisms with prokaryotic cells are called as Prokaryotes. • They are usually unicellular organisms . e.g. bacteria.

21.   EUKARYOTIC CELL   A typical Eukaryotic cell, as seen under light microscope has two major parts: 1. PLASMA MEMBRANE 2. CYTOPLASM and its ORGANELLES • The different substances that make up the cell are collectively called as Protoplasm.

22. PLASMA MEMBRANE It is a thin, elastic, semi permeable membrane of 7.5 to 10 nanometers thickness. It is a living membrane made up of phospholipid bilayer embedded with proteins. It is a flexible membrane and so it can fold in or out. It is made up of 55 % proteins, 25 % phospholipids, 13 % cholesterol, 4% lipids 3% carbohydrates. Functions of plasma  membrane • It protects the cell contents from the surrounding environment. • It provides a shape to the cell. • It is semi permeable and allows transport of certain substances in to and out of the cell. • It helps in forming cell junctions.

23. CYTOPLASM AND  ITS ORGANELLES • In eukaryotes, the protoplasm surrounding the nucleus is called cytoplasm . • It is a clear gelatinous fluid that fills the cell and surrounds the organelle. • It contains 90 % water , dissolved substances, minerals, sugar, irons, vitamins, amino acids, proteins and enzymes.

24. Cytoplasm > Is the seat for many pathways like glycolysis and HMP (Hexose mono phosphate) pathway. • The cytoplasm is also called as cytosol. • It contains well organized structures called organelles which vary in size from a few nanometers to many micrometers and they are specialized to carry out one or more vital functions of the cell.

25. The organelles  – Mitochondria –Endoplasmic reticulum –Golgi apparatus –Ribosomes –Lysosomes –Peroxisomes –Centriole –Vacuole –Nucleus

26. mitochondrion

27. These are tiny, sausage shaped structures of diameter 0.5 to 1 micrometre . It is called as POWER HOUSE OF THE CELL as it generates energy in the form of Adenosine Triphosphate (ATP) which is required by all cells. It is found both in plant and animal. It has a double membrane envelope: an inner membrane and an outer membrane .

28. The inner membrane is folded and pleated (double fold) and is called CRISTAE It provides large surface area for different biochemical processes as it contains many oxidative enzymes. Within this membrane are the proteins involved in electron transport chain, ATP synthase and transport proteins It is impermeable to molecules and ions but allows the free passage of carbon dioxide, oxygen and water.

29. The metabolites are transported across the membrane with the help of transport proteins. The outer membrane is a smooth phospholipid bilayer enveloping the mitochondria. It had enzymes like monoamine oxidase and NADH reductase. (NADH- Necotinamide adenine dinucleotide) An intrinsic protein called protein called porin form the channels that makes the membrane permeable to solutes and metabolites.

30. It allows the free passage of substances with no molecular weight less than 10,000 The membrane create two compartment… - THE SPACE BETWEEN THE OUTER AND INNER MEMBRANE IS CALLED THE INTERMEMBRANE SPACE. - IT IS HERE THAT OXIDATIVE PHOSPHORYLATION OCCUS. (REALISING ENERGY BY OXIDISE NUTRIENTS FOR ATP SYNTHESIS) - THE INNER CAVITY OF MITOCHONDRIA IS CALLED AS MATRIX. - IT IS PACKED WITH MANY ENZYMES LIKE PYRUVATE DEHYDROGENEASE, PYRUVATE ARBOXYLASE, ENZYMES FOR OXIDATION OF FATTY ACIDS, AMINOACIDS AND ENZYMES OF CITRIC ACID CYCLE. - IT ALSO CONTAINS MITOCHONDRIAL GENOME, MITOCHONDRIAL RIBOSOMES, tRNAs, dissolved oxygen, carbon dioxide and water.

31. MITOCHONDRIA FUNCTIONS: IT IS THE SEAT FOR KREB’S CYCLE (CITRIC ACID CYCLE) CONTAINS ENZYMES FOR OXYDATIVE PHOSPHORYLATION. MANY FATTY ACIDS ARE SYNTHESISED IN THE MATRIX CALCIUM CAN BE STORED IN THE MITOCHONDRIA AND RELEASED WHENEVER REQUIRED.

32. It is  a network of tubular and vascular structures extending from outer membranes of nucleus to the plasma membrane. It is seen to be spread throughout the cytoplasm and it provides a large surface area for various physiological activities. The inside of the vesicles and tubules is filled with an endoplasmic matrix. When the  ribosomes are attached on the outer surface of mebrane of the endoplasmic reticulum, it is called as Rough endoplasmic reticulum (RER). • It lies adjacent to the cell nucleus and its membrane is continuous with the membrane of the nucleus. • When there are no ribosomes attached to the endoplasmic reticulum, it is called as Smooth endoplasmic reticulum (SER).

33. Functions : –RER helps  in transporting proteins from ribosomes to golgi bodies. –Proteins that enters RER undergoes processing, folding and sorting –SER is involved in the synthesis of lipids, including cholesterol and phospholipids –In some cells, SER helps in the synthesis of steroid hoemones from cholesterol –In the cells of liver, SER helps in detoxifying drugs and harmful chemicals.

34. • It is  a stack of membranous sac, like a pile of disc. • It is present between endoplasmic reticulum and plasma membrane. • Like endoplasmic reticulum it is a single mebrane bound structure. • In animal cells it is present around the nucleus while in plant cell it is scattered throughout the cell. • These cells organelles which pack and sort the proteins before they are sent to their destinations.

35. Functions: –The proteins  that enter it from RER is modified, processed, sorted and transported in the form of vesicles to the plasma membrane and other destinations. –Glycolipids, sphingomyelin are synthesised within it. –In plant cells it help in the synthesis of polysaccharides needed by the cell wall. –It has role in synthesis of carbohydrates like galactose. –Primary lysosomes develop from mature Golgi bodies. –It plays an important role in lipid trafficking.

36. RIBOSOMES

37.   • They are  spherical shaped organelles seen either free in the cytoplasm or attached to RER. • They are found in eukaryotes and prokaryotes. • They are synthesized by nucleolus. • The ribosomes link the amino acids together in the order that is specified by the messengers RNA. • They are made up of two subunits - a small sub unit and a large sub unit. • The small  sub unit reads the mRNA while the large sub unit assembles the amino acids to form large polypeptide. • The ribosomal sub units are made up of one or more eRNA and proteins. • Function:- –It is the site for protein synthesis.

38. LYSOSOME

39. • They are  tiny sac like organelle of size 0.5 to 1.5 µm, which are membrane bound and found in hudreds in a single cell. • They are formed from golgi bodies as small vesicles which bud off from them. • Within the sac there are several hydrolytic enzymes that breakdown macromolecules like nucleic acid, proteins and polysaccharides. • They are  also called as “suicidal begs” as enzymes contained in term can digest the cell's own material when damaged or dead. • The important enzymes in it are DNA- ase , RNA- ase , protease, lipase,glycosidase , phosphatase, sulphatase which are synthesised in the endoplasmic reticulum and then transported to the golgi bodies.

40. Functions: –It helps  in digestion of food releasing enzymes. –They digest worn out organelles –It helps in defence by digesting germs –It helps sperm cells in entering the egg by breaking through the egg membrane –It provides energy during cell starvation by digestion of cells own parts.

41. PEROXIXOMES

42. • It is  a type of microbody which is small, spherical shaped, with single membrane and of size 0.5-1.5 µm. • They are found in both animal and plant cells. • Just like lysosomes they contain many enzymes which help in biological reactions. • They are formed from endoplasmic reticulum unlike lysosomes which are formed from golgi bodies. Functions: –The enzymes  found in peroxisomes are usually used for different metabolic reactions and for digesting different materials in the cell. –They help in oxidation of many substances resulting in the formation of hydrogen peroxide as a by product. –But, it contains enzymes peroxidase or catalase which decomposes this harmful hydrogen peroxide into water and oxygen or uses it to oxidize other organic compounds like phenol, alcohol, formaldehyde, etc.

43. –They are also  involved in catabolism of fatty acids (beta oxidation), D- aminoacids and polyamines. –They are needed in the synthesis of plasmalogens (type of ether phospholipid needed for functioning of brain and lungs.) –They participate in synthesis of cholesterol, bile acids and myelin. –In plants it helps in photorespiration and symbolic nitrogen fixation.

44. • They are  long, hollow cylindrers of size 24 nm in diameter and can grow up to a length of 50 µm. • They are found in eukaryotic cell. • They are  made up of two globular proteins namely α-tubulin and β-tubulin. • Along the microtubule axis tubulins are joined end to end to form protofilaments.

45. CYTOSKELETON

46. • They are  important component of cytoskeleton and so they found throughout the cytoplasm. • They are organised by microtubule organising structures, primarily the centrioles. Functions: –They are  part of cytoskeleton and so provides mechanical support to the cell –It helps in organisation of cytoplasm –They help in segregation of chromosomes during mitosis –They are used for locomotion (movement from one place to another) when present in flagella and cilia)

47. • They are  small rod like structures of size 4-7nm in diameter found in the cytoplasm of all eukaryotic cells, forming a part of the cytoskeleton. • They are made up of protein Actin (contractile protein) • Functions:- –It provides support and shape to cell –Along with myosin it helps in contraction –It helps in cytokinesis ( a physical process of cell division)

48. • They are  part of the cytoskeleton in the cytoplasm found surrounding the nucleus and extending to the plasma membrane. • They are made up of different types of fibrous proteins unlike microtubules which are made up of actin. • They are of size 8-12 nm in diameter. • They are found in hair, nails, scales and skin since they have high tensile strength. • Due to  their rope like structure they provide mechanical strength to the cells and help cells with stand stress like stretching and changing shape. Eg. Keratin filaments in skin and epithelial cells.

49.   • It is  cylindrical in shape and of length 0.5 micrometer. • it is present in all animal cells just outside the nucleus. • It does not have a membrane. • All centrioles are made up of protein strands called tubulin. • Each centriole has 9 sets of inter connected peripheral tubules and each set has 3 micro tubules arranged at definite angles making the shape of a cylinder.

50. • It has  its own DNA and RNA and therefore it is self duplicating. • Two centrioles when oriented at a right angle, forms centrosome. Functions: It is involved in cell division. They are seen in the process of both meiosis and mitosis. It helps in the formation of cilia and flagella. It helps in organization and alignment of microtubules within the cell.

51. • It is  a membrane enclosed fluid filled sac present in animal and plant cells including fungi. • It contains organic and inorganic molecules within it. • They do not have particular size or shape but adjust themselves according to the need of the cell.

52. Functions: It helps in  removing waste products from the cell It isolates substances that are harmful to the cell It holds water and waste products within it It helps in maintaining the internal pH of cell It helps to maintain hydrostatic pressure within the cell It plays a major role in autophagy by maintaining a balance between biogenesis and degradation

53. NUCLEAR ENVELOPE or NUCLEAR MEMBRANE

54. • It is  a specialised double membrane- bound protoplasmic body present at the centre of the cell. • It is known as the cell's information center as it houses the chromosomes. • The double membrane around the nucleus is called nuclear membrane or nuclear envelope. • It is made of proteins and lipids, • It enclose the nucleus to keep it separate from surrounding materials off the cell. • The outer  membrane is continuous with endoplasmic reticulum and it has ribosomes attached on the outer surface. • It has several large nuclear pores through which nuclear transport of large molecules, small molecules and ions occur. • The space between the nuclear membranes is called perinuclear space and it is continuous with the lumen of rough endoplasmic reticulum.

55. • Within the  nuclear membrane is a jelly like substance called karyolymph or nucleoplasm. • Within it there is a network of chromatin fibrils which condense to form chromosomes during cell division • The nucleolus is present within the nucleus. • It does not have a membrane around it. • It synthesizes rRNA and assembles. • It regulates the synthetic activity of nucleus.

56. Functions: It controls the  hereditary charateristics of an organism Protein synthesis, cell division, growth and dfferentiation occurs in it Stores heredity materials in the form of DNA It is a site for transcription in protein synthesis Nucleolus helps in the synthesis of ribosomes It regulates the integrity of genes and gene expression

57. A prokaryote  is a single-celled organism that do not have a 'true nucleus’. ( Eg. Bacteria, Archaea.) • It does not contain any membrane bound organelles like mitochondria, nucleus, endoplasmic reticulum etc. • They have many ribosomes scattered throughout their cytoplasm and nucleoid which contains the DNA.

58. Parts of prokaryotic  cell : • Flagellum : It is long whip like structure that helps in locomotion • Pili : Small hair like structure present on the surface which helps in attaching to the surface of other bacterias .

59. Cell membrane: –It  surrounds the cytoplasm and regulates the flow of substance in and out of cell Capsule : It is a polysaccharide layer that is outside the cell envelop. It enhances the ability of bacteria to cause disease Cell wall : It  is the outer most covering of the cell and it gives shape to the cell Cytoplasm : It is gel like substance present within the cell. It contains enzymes, ions, organic molecules, ribosomes, nucleoid

60. Ribosome: It is  organelle which helps in protein synthesis. Nucleoid : It contains the genetic material Plasmid : - A small DNA  molecule within the cell that can replicate itself. –They are small double stranded and circular in shape. –They are usually found in bacteria.

61. MICROSCOPY > It is  the technical field of using microscopes to view samples and objects that cannot be seen with the unaided eye. It is the science of investing small objects using microscopes. MICROSCOPE Microscope is  an instrument used to see objects that are too small for the naked eye. • Identification of minute organisms are necessary for diagnosis and treatment.

62. MICROSCOPE

63. TYPES OF MICROSCOPE  Optical  light microscope  Electron microscope  Dark field or ultra-microscope  Phase contrast and differential interference contrast microscope  Fluorescent microscope  Ultraviolet microscope

64. OPTICAL/LIGHT MICROSCOPE  It a  type of microscope which uses visible light and a system of lenses to magnify images of small samples.  It was invented by Hans Janssess and Zacharias in 1550.

65. The eyepiece, objective  lenses, reflector, condenser and stage is first cleaned by a tissue. The specimen slide is then placed on the stage with help of clips at the center. Observe through the eyepiece and focus the object with the help of coarse or fine adjustment. The reflector can be adjusted to get proper light. The light shining  through the specimen is focused by the lense so that a magnified image can be seen through the eyepiece.

66. ELECTRON MICROSCOPE  It  was designed by Ernst Ruska and Max Knoll in 1932 in Germany. It is a type of microscope that uses a beam of electrons to illuminate a specimen and produce a magnified image. The wave length  of lens as when compared to light is smaller and so they helps to magnify even very small objects. It has high magnifying power when compared to light microscope as electrons have shorter wavelength than visible light. It uses electrostatic and electro magnetic lenses to control electron beam and focuses in to form an image. It is used to see micro organism, cell organelles, large molecules, biopsy samples, etc.

67. TYPES OF ELECTRON  MICROSCOPE Transmission Electron microscope Scanning Electron microscope Reflection Electron microscope Scanning Transmission electron microscope Low voltage electron microscope

68. Transmission Electron Microscope It  uses high voltage electron beam to create an image. The beam of electron is sent through the specimen. Some electrons are reflected while others pass through it creating an image of the specimen.

69. Scanning Electron Microscope It  does not produce a complete image of the specimen. It scans the surface of the specimen and forms an image by detecting electron that are reflected or absorbed. Reflection Electron Microscope: Similar TEM, the reflected electrons are detected to get the information about the surface of the specimen.

70. Scanning Transmission Electron Microscope It  combines high magnification of TEM with surface details of SEM. It helps to perform a complex analysis of the specimen. Low Voltage Electron Microscope It operates at accelerating voltage of a few kilo electrovolts or less.

71. DARK MICROSCOPE It  is a microscope which has special consider with stop disc to scatter the light as it reaches the specimen. Here reflected light is used in place of transmitted light. The oblique beams of refracted and detracted light coming from the sides passes into and over the specimen to illuminate it.

72. The object looks  bright in the dark background. It is useful in observing small living objects and small organelles like nucleus, mitochodria , vacuole, etc. It is usally used to see unstained objects.

73. 4. PHASE CONTROL  MICROSCOPE   It is a microscope which helps to see unstained micro organisms.  It is the technique that converts phase shifts in light passing through a transparent specimen to brightness changes in the image.  The light passing through two different materials with different refractive index will undergo a change in the phase of light.  These phase differences  are converted to difference in intensity of light, making image appear dark against a light background.  Phase contrast microscopy improves the contrast and make the structures visible.

74. 5. FLUORESCENT MICROSCOPE It  is an optical microscope that uses fluoresce and phosphorescence to study the properties of organic and inorganic substances. Special dyes like fluorescein, rhodamine and auramine are used.

75. 6. ULTRAVIOLET MICROSCOPE    It  is a microscope that has quartz lens and slides uses ultra violet light as illumination instead of common light.  The shorter wavelength (180-400 nm) of ultraviolet rays compared to common light, brings about higher resolution of objects.

76. CELL FRACTIONATION • It  is the technique of rupturing the cell to separate various cell components while preserving their individual functions in order to study their structure and chemistry. • Cells can be broken down by many ways like subjecting it to osmotic shock or ultrasonic vibration or ground in a small blender.

77. • These procedures  break up the cell in to fragments and nuclei, mitochondria, golgi body, lysosomes, peroxisomes, etc. can be separated. • The cells are first suspended in 0.25 molal sucrose solution at 0-4℃. • Cells are then ground well to form an isotonic slurry called homogenate. • The homogenate is then subjected to different values of centrifugal force. This helps  in separating the cell components by size and density. • At relatively a low speed the large components like nuclei sediment to form a pellet at the bottom of the centrifuge tube, at slightly higher speed, a pellet of mitochondria is formed, and at even higher speeds and with longer period of centrifugation, first the small closed vesicles and then the ribosomes can be collected.

78. GEL ELECTROFORESIS It  is a method of separating macromolecules like DNA,RNA and proteins based on their size and charge, by passing it through a gel medium namely agarose and by applying an electric field. • The smaller molecules move faster than the larger ones through the pores of the gel and the molecules in the gel can be stained to make them visible. • Ethidium bromide  is the most commonly used stain to make DNA or RNA strands visible.

79. CHROMATOGRAPHY • It is  the technique used for separation of a mixture by dissolving it in a solution or suspension and allowing it to pass through a medium in which the components move at different rates. • In this technique there is a stationary phase and mobile phase. • The various constituents of the mixture travel at different speeds, causing them to separate.

80.   TYPES OF CHROMATOGRAPHY Column  chromatography Paper chromatography Thin layer chromatography Gel filtration chromatography  Iong exchange chromatography Affinity chromatography