Week 9 Myeloma & Lymphoma (3).docx PDF

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Lindsay McCrea...

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1. Streaking for isolation—SFIC a. **When streaking, always label the bottom plate or where the microorganisms are.** i. Labeling the lid can be useless as the lid can be lost b. Streaking for isolation allows for use to dilute bacteria so we get individual bacteria colonies c. Use aseptic technique (do not take off the lid and set it down, don’t put the plate up near your face. d. Use your lid to shield airborne microbes. e. This is a dilution technique and it is critical to flame (and cool) yourloop in between each zone. Want individual colonies to make smears or inoculate culture tubes/plates. 2. Gram stain or Gram reaction a. Gram positive i. Fixation - white ii. Crystal violet - Blue iii. (Mordant) Iodine Treatment - violet iv. Decolorization - violet v. Counterstain safranin - violet b. Gram Negative i. Fixation - white ii. Crystal violet - Blue iii. (Mordant) Iodine Treatment - violet 1. Iodine helps with stain retention iv. Decolorization - white v. Counterstain safranin - pink

c. 1. Make smear. Make multiple smear slide (Fixation) d. 2. Add for 1 min and rinse gently with water (Crystal Violet) e. 3. Add for 1 min and rinse gently with water (Iodine Treatment)

f. 4. Add 3-6 drops of 95% almorphcohol and rinse gently with water (Decolorization) g. 4. Add for 30 seconds and rinse gently with water (Counterstain safranin) 3. **Negative staining** using the acidic stain/dye nigrosin or India Ink a. 1. Add small drop of nigrosine onto 1 end of slide b. 2. Aseptically transfer 1 loopful of Negative stain broth onto slide c. 3. Use a 2nd slide at 45 degree angle to smear mixture across the slide. d. 4. Allow to air dry. e. 5. Look under a microscope. 4. Shape and morphology a. Coccus b. Bacillus c. Vibrio i. Spirals/ comma shaped 5. Acid fast stain - (Basic Stain)

a. Acid fast positive - pink b. Acid fast negative - dark blue/purple i. Use of Acid Fast Stain is **because of poor results from Gram stain** ii. Identify acid-fast organisms-genus Mycobacterium (myolic acid) 1. Example: Mycobacterium Tuberculosis, Mycobacterium Leprosy, Mycobacterium Kansasii a. Resistant to antibiotics b. Treated with isoniazid (INH) iii. Characterized by wax-like, nearly impermeable cell walls iv. Made up of mycolic acid and large amounts of fatty acids, waxes, and complex lipids v. Slow growing (Fast-growers 2 weeks Slow growers 6 weeks, non-mycobacterium 12-18 hours) vi. Resistant to most compounds and require a special staining technique. vii. Steam helps to loosen up the waxy layer and promotes entry of the primary stain inside the cell c. **This stain require the use of steam heat**

(Steam heat)

6. Spore stain a. **requires steam heat** i. Resistant to conventional staining procedures ii. **Steam heat facilitates stain entry of malachite green (primary stain) & allows for the staining of spore itself** iii. Steam heat to loosen the bonds between calcium, dipicolinic acid and glycerol b. It is important to determine if a bacterium is a spore former or not (non-spore former) for identification and for treatment and control. c. **Spores are resistant to heat, desiccation, chemicals, and radiation** i. When radiation goes away, they go back to a vegetative state d. Normal growing cells are called vegetative cells or non-spore formers while spore producing species are called sporeformers. e. After staining, vegetative cells red in color and green. f. **If spore in cell, it’s an endospore and if spore inside cell, it’s called a free spore** g. Again, staining cells for the presence or absence of structures will allow them to identify these bacteria. h. Endospore Position characteristic of some species Bacillus spp. central spore Clostridium spp. Subterminal and terminal

i. Terminal endospore

j. Sub central/ Sub terminal k. Central (middle)

● Spore Stain Protocol ○ The students on one side of a bench will perform a spore stain first. ○ Aseptically prepare smears of Bacillus sp. and E. coli on one slide. Be sure to air dry and heat fix. ○ Place the slide over a steam bath and place a small piece of pre-cut paper towel over their smear. ○ Put a few drops of malachite green (a basic stain) on the paper towel and as it dries, add more. Do not allow the stain to get into the water! If it happens, it does…STOP. REMOVE BEAKER FROM HEAT. CONTACT INSTRUCTOR. ○ Keep the paper towel moist for 5 minutes. You will need to be diligent and watch your slide. ○ After 5 minutes, use forceps to remove the paper towel and place in the empty Petri dish. ○ Take slide to the staining trays, and gently rinse with water. ○ Add a few drops of safranin to your smear. Let that stain (another basic stain) sit for one minute. ○ Gently rinse with water, dry with bibulous paper, and view at 100X with immersion oil.

● Arrows point to free spores, vegetative cells and endospores (top to bottom). Green spores. Red vegetative cells.

● Terminal or subterminal spores 7. Capsule Stain a. Concerned about capsule-formers for several reasons, such as their resistance to antibiotics. i. Can evade immune system b. Capsule stain-**requires no heat fixation** i. **Negative stain also require no heat fixation** c. Level of delicacy needed as conventional staining procedures. d. Things unique to Capsule stain i. Heat fixation would cause a capsule to shrink and “dissolve”, and thus, it might not be visible. ii. The primary stain is crystal violet and because capsules are nonionic (No charge), they do not pick up the stain, but the bacterium will. 1. **Typical bacteria carries a negative charge so we need a positive stain** iii. Both the decolorizer and the secondary stain are 20% copper sulfate iv. The capsules do not look copper blue, rather are light, light, light, light

blue which really means white. 8. Capsule Stain Protocol a. The other side of the bench will perform capsule staining. b. Aseptically prepare smears of Klebsiella pneumoniae (capsule former) and E. coli on one slide. Be sure to air dry and DO NOT heat fix. c. Heat fixation would cause a capsule to shrink and “dissolve”, and thus, it might not be visible. d. Add a few drops of crystal violet (basic stain) to the smear (again air dry no heat fixation) and let sit for 1-2 minutes. e. Gently rinse with the copper sulfate (drip, drip, drip) and then add drops of copper sulfate and allow to sit for 3-5 minutes. f. The best thing to do is dry with bibulous paper and view at 100X with immersion oil. Then, groups change staining stations. “Spore former folks” will now do a capsule stain and vice versa.

9. 10. Chapter 4: pg 86 a. Pg 100: 1, 3, 5, 6, 9 i.

(1) 1.5 mg/ 0.75 mg x 1 tablet = 2 tab

ii.

(3) 500 mg/ 250 mg x 1 cap = 2 capsules

iii.

(5) 650 mg/ 325 mg x 1 tab = 2 tab

iv.

(6) 20 mg/ 10 mg x 1 cap = 2 capsules

v.

(9) 0.5 mg/ 0.25 mg x 1 scored tab = 2 scored tab

b. Pg 110-111: 2,7,8,12,16 i.

(2) 0.25 mg/ 0.5 mg x 1 scored tab = 0.5 scored tab

ii.

(7) 10 mg/ 2.5 mg x 1 tab = 4 tablets

iii.

(8) 200,000 units/ 400,000 units x 1 scored tab = 0.5 scored tab

iv.

(12) 0.3 mg/ 0.1 mg x 1 tab = 3 tab

v.

(16) 0.625 mg/ 1.25 mg x 1 scored tab = 0.5 scored tab .

c. Pg 117: 1, 2, 4, 5 i.

(1) 750 mg/ 250 mg z x 1 mL = 3 mL

ii.

(2) 500 mg/ 250 mg x 5 mL = 10 mL

iii.

(4) 150 mg/ 100 mg x 1 mL = 1.5 mL

iv.

(5) 5 mg/ 10 mg x 1 mL = 0.5 mL

d. Pg 119: 6, 7, 8, 9, 10 i.

(6) 0.02 mg/ 0.05 mg x 1 mL = 0.4 mL

ii.

(7) 30 mEq/ 20 mEq x 15 mL = 22.5 mL .

iii.

(8) 0.25 mg/ 0.25 mg x 1 mL = 1 mL

iv.

(9) 3 mg/ 1 mg x 1 mL = 3 mL

v.

(10) 12.5 mg/ 6.25 mg x 5 mL = 10 mL

11. Ch 5: pg 142, 164-168 a. Pg 157: self-test 2

i. Ratio

? g per? mL

? g = ? mL

? g/? mL

1:20

1 gram per

1 g = 20 mL

1g/20 mL

2 g = 15 mL

2 g/15 mL

1 g = 500 mL

1g/500mL

20mL

2:15

2 gram per 15 mL

1:500

1 gram per 500 mL

2:2000

2 gram per 2000 2 g = 2000 mL

2g/2000mL

mL

1:4

1 gram per 4

1 g = 4 mL

1g/4mL

2 g = 25 mL

2g/25mL

4 g = 50 mL

4g/50mL

mL

2:25

2 gram per 25 mL

4:50

4 gram per 50 mL

1:100

1 gram per 100

1 g = 100 mL

1g/100mL

3 g = 75 mL

3g/75mL

mL

3:75

3 gram per 75 mL

5:1000

5 gram per 1000 5 g = 1000 mL mL

ii. Ways to treat anemia: -

-

Blood transfusions -

Whole blood

-

Packed red blood

-

Fresh frozen plasma

Other blood products that can be transfused -

Platelets

-

Clotting factors

-

Albumin

Blood transfusions:

5g/1000mL

-

Whole blood - for injuries and critical stages

-

Packed red blood cell - the RBC from the whole blood (for patient’s w/ anemia) -

Even use these in surgeries

Blood cancers: (Myeloid (bone marrow) vs. lymphoid (lymph tissue)) -

Chronic vs acute

-

Blood cancers (excess production of blood cells (RBC, WBC, and platelets) that exceed normal production -

-

Doesn’t serve any purpose to fight an infection, form clot carry more oxygen

Blood cells grow on their own at various stages of hematopoiesis -

Can have excess immature cells stuck dividing in bone marrow or in bloodstream

Myeloid cells: -

Are rbc, platelets, and granulocytes

-

AML - acture immature myeloid blast cells in bone marrow -

-

Found in adult patients

CML - chronic excess production of platelets, RBCs, and granulocytes -

Precursor for aplastic anemia (affects all types of blood production)

-

Found in elderly patients

Lymphoid cells: (leukemia) -

B cells and T cells

-

ALL- acute immature B cells and T cells in bone marrow (bruising everywhere)

-

-

Most common leukemia in children

-

High cure rate and survival ~90%

CLL - chronic mature b cells and t cells -

Not fighting infection

-

Found in elderly patients

myeloma - cancer of plasma cells/ b-cells, antibodies (multiple myeloma) -

Exculsively in adults, usually men >40

-

Feel like “CRAB” -

C - calcium elevation in the blood

-

R - renal failure

-

Plasma cells produce light chain antibody fragments (bensjones) that are produced in excess and accumulate in kidney leading to renal failure

-

A - anemia -

-

Overcrowding of blood cells that make rbc not able to be produced usually

B - bone lesions and bone pain -

Plasma cells form tumors w/in bone marrow that spread into damaged bone tissue and cause loss of calcium in bones. When calcium leaves, it leaves behind osteolytic lesions.

Nursing considerations for blood cancer patients: -

Immunocompromised precautions (neutropenic precautions or reverse isolations) -

No fresh flowers

-

Neutropenic diet

-

Good hand hygiene

-

Wear PPE: gloves, gowns, masks

-

Limit exposure to visitors w/ respiratory infections

Lymphatic System

● 2 main functions ○ 1.) Return excess fluid to our venous blood ■ Maintain fluid balance, blood volume, blood pressure ○ 2.) Transport and house cells from our immune system ■ Lymphocytes (Leukocytes) and other immune cells

● Anatomy ○ Lymph vessels ■ Function ● Return fluid to our venous blood (Veins) ○ Interstitial fluid that made it into the lymph capillary (Lymph) ○ Lymphatic tissue and organs ■ Organs ● Support our immune system ● Provide the support by transporting and housing immune cells

● Lymph and Lymph Capillaries ○ Lymph capillaries -

■ Take up extra interstitial fluid ■ Made up of single cells endothelial layers that make up the wall ■ Have “NO” basement membrane that surrounds itself like regular capillaries ■ Lumen is larger than in regular capillaries ■ Endothelial cells are overlapping ■ Hydrostatic pressure drive fluid into the lymph capillaries ● Pressure increases and flap close so fluid can’t escape ■ Blind ended (closed end on one side) ○ Lymph ■ Made up of water, dissolved solutes, ions, hormones, small amount of protein from capillaries, cellular debris, pathogens (Cancer) ● Not the same as our blood plasma, but contain about the same components

● **Lymphatic capillary -> Lymphatic Vessels -> Lymphatic Trunk -> Lymphatic Duct**

● Lymphatic vessels, trunks, and ducts ○ Lymph capillaries are going to merge into the lymphatic vessels ■ Facilitating of return of fluid that is not reabsorbed ■ Have 3 tunics (layers) ● Smooth muscle in the tunica media ■ Have valves

● Insure one way flow ■ Skeletal and respiratory pump ■ Arterial movement ○ Lymphatic vessels merge into the Lymphatic trunk ○ Lymphatic trunk merge into the lymphatic duct (Lymph Duct) ■ Largest of the lymph vessels ■ Responsible for dumping that lymph back into our lymph system

● Lymph Duct (Right and Thoracic) ○ Right lymphatic duct - (GREEN)

■ Drain the green part the body ■ Dump the lymph into the vein ○ Thoracic duct - (RED) ■ Drain the whole rest of our blood ■ Bring blood and dump blood in the superior vena cava ○ Cisterna chyli ■ Collect lymph from abdominal and intestinal area ○ Lacteal ■ Collecting lymph that contains dietary lipids

● Lymphatic Tissues and Organs ○ Plays a role in supporting our immune system ○ Primary Lymphatic Structure ■ Thymus ■ Red bone marrow ■ **Formation and maturation of lymphocytes** ○ Secondary Lymphatic structure ■ Lymph nodes ■ Tonsils ■ Spleen ■ MALT ■ **House lymphocytes and other immune cells** ■ Where immune response is initiated

● Red bone marrow ○ Primary lymphatic structure ○ Site of hematopoiesis ○ A place where our B-cells mature ○ T-cells mature in the thymus ○ B-cells and T-cells play a role in our immune system

● Thymus ○ Site of T-cell maturation ■ Formed in the bone marrow but mature in the thymus ○ Thymus largest during adolescence but start to atrophy (grow smaller) with age

● Lymph Nodes - (Secondary lymphatic structures) ○ Often found in clusters ○ Functions: ■ Cleansing lymph: ● Act as filters ● House macrophages that remove and destroy microorganisms and debris that enter lymph

■ Immune system activation: ● Offer a place for lymphocytes to become activated and mount an attack against antigens

● Lymph flow through afferent vessels into the lymph nodes (multiple afferent vessels) ● One efferent vessels where lymph drains ○ When drained, lymph should be cleansed of debris

● Tonsils ○ Simplest lymphoid organs ○ Tonsils’ function is to gather and remove pathogens in food or air ○ Tonsillar crypt ■ Trap pathogens

● Spleen ○ Largest of lymphatic organs ○ Does not filter lymph, only filters blood ○ Functions: ■ Site of lymphocyte proliferation and immune surveillance and response ■ Cleanses blood of aged blood cells and platelets; macrophages remove debris ○ Has Sinusoid capillaries

● MALT - (Mucosa Associated lymphoid tissue) ○ Found in mucosa (mucous membrane) of respiratory tract, genitourinary organs, and digestive tract

○ Peyer’s patches ■ Collection of lyphoid tissues in our intestines...