8.

Immediately cut the entire small intestines and

place it on a plastic board with a plastic ruler
underneath.
9. Measure the distance travelled by the charcoal
Materials suspension.
• Dissecting sets 10. Record, note and compare the results obtained.
• Plastic board
• Gloves 3 Animal Groups
• Tuberculin syringe • Positive Control Groups
• Intubation needles o Atropine sulfate - Mouse #1
• Plastic rulers o Neostigmine - Mouse #2
• Negative Control Group
Test Animals o Normal saline solution - Mouse #3
• Albino mice
Test Animal No. Group Weight (g)
Drugs 1 (+) control – 32
• Atropine Sulfate – 5 ug/mL Atropine SO4
• Neostigmine methyl sulfate – 0.02 mg/kg BW 2 (+) control – 38
• Normal Saline Solution Neostigmine
• 2% charcoal suspension – 0.3 to 0.5 mL/20 g 3 (-) control – NSS 35

Procedures Computation for Atropine SO4

1. Prepare all the materials needed. Dose Computation
2. Weigh all the test animals and divide them into • Positive Control Group
three (3) groups. (One group will be treated with o Atropine sulfate (5 ug/mL)
Atropine sulfate, one for Neostigmine and the ▪ Ideal volume limit for
remaining group will be given Normal saline Intraperitoneal = 5-10 mL/kg
solution) BW
3. Compute for the dose of the drugs to be ▪ Weight of Mouse #1 = 32 g =
administered to the test animals. 0.032 kg
4. To the first group, administer Atropine Sulfate Volume Limit (Using the standard 10 mL/kg BW)
intraperitoneally (IP); to the second group, 10 𝑚𝐿 𝑥
=
Neostigmine; and to the third, Normal saline 1 𝑘𝑔 0.032 𝑘𝑔
solution. x = 0.32 mL volume of drug to be administered to the
5. After 30 minutes administer an appropriate 32 g mouse
amount of 2% charcoal suspension in sterile This is confirmed using
distilled water. 1 𝑚𝐿 𝑥
=
6. Fifteen minutes after administration, sacrifice the 100 𝑔 32 𝑔
x = 0.32 mL
animals by cervical dislocation.
7. Open the peritoneal cavity and observed the
movements of the intestines.
Dose (mg) ▪ Using 0.5 mL/20 g, the volume
• Convert: 5 ug = 0.005 mg of 2% charcoal to be
0.005 𝑚𝑔 𝑥 administered orally is:
=
1 𝑚𝐿 0.32 𝑚𝐿 0.5 𝑚𝐿 𝑥
x = 0.0016 mg of drug to be administered to the =
20 𝑔 38 𝑔
32 g mouse x = 0.95 mL of 2% activated charcoal to be administered
to the 38 g mouse
o 2% Charcoal Suspension NOTE: Although the standard volume limit for oral
(0.3 to 0.5 mL/20 g) gavage is 10 mL/kg BW for mouse, use the specific
▪ Using 0.5 mL/20 g, the volume volume limit for 2% activated charcoal that is provided
of 2% charcoal to be protocol in the experiment.
administered orally is:
0.5 𝑚𝐿 𝑥 Computation for NSS (Normal Saline Solution)
=
20 𝑔 32 𝑔 • Negative Control group:
x = 0.8 mL of 2% activated charcoal to be administered
o Normal Saline Solution (NSS)
to the 32 g mouse ▪ Ideal Volume Limit for
Intraperitoneal = 5-10 mL/kg
NOTE: Although the standard volume limit for oral BW
gavage is 10 mL/kg BW for mouse, use the specific
▪ Weight of Mouse #1 = 35 g
volume limit for 2% activated charcoal that is provided
= 0.035 kg
protocol in the experiment. Volume Limit (Using the standard 10 mL/kg BW)
10 𝑚𝐿 𝑥
Computation for Neostigmine =
1 𝑘𝑔 0.035 𝑘𝑔
Dose Computation x = 0.35 mL volume of NSS to be administered to the
• Positive Control group: 35 g mouse
o Neostigmine (0.02 mg/kg BW)
▪ Ideal Volume Limit for Dose (mg)
Intraperitoneal = 5-10 mL/kg • NOT APPLICABLE
BW o NSS is only a vehicle!
▪ Weight of Mouse #1 = 38 g o No relevant therapeutic effect to the
= 0.038 kg study!
Volume Limit (Using the standard 10 mL/kg BW) o You don't need to compute using 0.9%
10 𝑚𝐿 𝑥 concentration because you will use NSS
=
1 𝑘𝑔 0.038 𝑘𝑔 as is! (No mixing or compounding
x = 0.38 mL volume of drug to be administered to the
involved!)
38 g mouse
Dose (mg) o 2% Charcoal Suspension
0.02 𝑚𝑔 𝑥 0.5 𝑚𝐿 𝑥
= =
𝑘𝑔 0.038 𝑘𝑔 20 𝑔 35 𝑔
x = 0.00076 mg of drug to be administered to the x = 0.875 mL of 2% activated charcoal to be
38 g mouse administered to the 35 g mouse
o 2% Charcoal Suspension
(0.3 to 0.5 mL/20 g)

Test Vol. of Vol.

Weight Distance travelled by
Animal Group Dose (mg) Charcoal admin
(g) charcoal
No. (mL) (mL)
1 (+) control - Atropine SO4 32 0.0016 0.8 0.32 Shortest (eg 220 mm)
2 (+) control - Neostigmine 38 0.00076 0.95 0.38 Longest (eg 450 mm)
3 (-) control - NSS 35 N/A 0.875 0.35 Intermediate (eg 370 mm)
Conclusion
• Atropine is a muscarinic antagonist = increased
peristaltic relaxation = shorter distance traveled
by charcoal suspension = Spasmolytic
Agent/Antispasmodic Agent
• Neostigmine is a muscarinic agonist = increased
peristaltic contraction = longer distance traveled
by charcoal suspension
• NSS = no known muscarinic activity = normal
peristaltic activity = normal distance traveled by
charcoal suspension
Use of Activated Charcoal in the Experiment
• Transit/intestinal motility indicator
• To determine the transit or intestinal motility of
mice after being exposed to Atropine SO4,
Neostigmine, and NSS, respectively
- layers in the adrenal glands
• outer layer: adrenal cortex
• inner layer: adrenal medulla
- kidneys are connected to the ureter, bladder, and urethra
- renal pelvis: point that interconnects the ureter with the
kidneys
Types of Nephrons
• Cortical nephrons
o Located entirely in the cortex
o Includes most nephrons
• Juxtamedullary nephrons
o Found at the boundary of the cortex and
medulla

Nephron
- site of action of diuretics
- The functional filtration unit in the kidney
- above the kidneys, we have adrenal glands and
suprarenal glands
- In both kidneys: approximately 2.5 million nephrons
- Are microscopic: measure about 5 centimeters in
length
NOTE: Right kidney ~2cm lower than left
• Due to the larger size of liver on the right side of
the body

- nephron’s segments/portions
• Glomerulus (covered by the Bowman’s capsule)
• Proximal convoluted tubule
• Loop of Henle
• Distal convoluted tubule
• Collecting tubule (papunta sa ureter)
Collecting duct – farther from the glomerulus, unlike the
- layers in kidney collecting tubule
• outer layer: renal cortex
• inner layer: renal medulla
 Segment
Glomerulus
Proximal Convoluted Tubule (PCT)
Proximal tubule, straight segments
Thin descending limb of Henle’s loop
Thich ascending limb of Henle’s loop (TAL)
Distal Convoluted Tubule (DCT)
Cortical Collecting Tubule (CCT)
Medullary collecting duct
Functions
- Formation of glomerular filtrate
- Reabsorption of 65% of filtered Na+/K+/Ca2+, and Mg2+; 85% of NaHCO3,
and nearly 100% of glucose and amino acids.
- Isosmotic reabsorption of water.
- Secretion and reabsorption of organic acids and bases, including uric
acid and most diuretics
Passive reabsorption of water
Active reabsorption of 15-25% of filtered Na+/K+/Cl-; secondary
reabsorption of Ca2+ and Mg2+
Active reabsorption of 4-8% of filtered Na+ and Cl-; Ca2+ reabsorption
under parathyroid hormone control
Na+ reabsorption (2-5%) coupled to K+ and H+ secretion
Water reabsorption under vasopressin control

Diuretics 4. Osmotic diuretics

• lower blood pressure by reducing the blood 5. Carbonic anhydrase inhibitors
volume and reduction in body sodium 6. Sodium/glucose cotransporter (SGLT2) Inhibitors
o can also be used for intracranial pressure, 7. Vasopressin (ADH) Antagonists
glaucoma (intraocular pressure) • ADH – anti-diuretic hormone (secreted by
o kung saan may sodium, may water din the posterior pituitary gland along with
• the first agents tried in mild hypertension oxytocin)
• cause electrolyte and acid-base disturbances • Also called Arginine Vasopressin
• can also be used for edema, CHF, and others - first 5 types of diuretics are the most popular

Types: Others:
1. Thiazides • Na-H exchanger antagonists
2. Loop diuretics • Arginine vasopressin receptor 2 antagonists
3. Potassium-sparing diuretics • Xanthines
 Type Definition/MOA Uses and Adverse Effects
THIAZIDE DIURETICS
Thiazide - Chemically related to sulfonamides Uses:
• Bendroflumethiazide - Prototype: • Mild or moderate HTN
• Benzthiazide • Hydrochlorothiazide • Congestive heart failure (CHF)
• Chlorothiazide - Major site of action: o CHF = fluid builds up around the
- only parenteral thiazide (IV) • Distal convoluted tubule heart and causes it to pump
- also commonly used inefficiently
• Polythiazide MOA: • Liver and renal disease
• Trichloromethiazide - inhibit NaCl reabsorption from the o Ascites (abnormal buildup of
• Hydrochlorothiazide (HCTZ) luminal side of epithelial cells in the distal fluid in the abdomen; can be
- most popular and commonly used convoluted tubule by blocking the caused by liver failure/liver
• Hydroflumethiazide Na+/Cl- transporter = Modest increased cirrhosis)
NaCl excretion o Nephrolithiasis (kidney stone
Thiazide-like formation)
- chemically known as sulfonamides Other effects: • Hypercalciuria (excess calcium in the
- structurally similar with sulfonamide - Potassium wasting urine)
antibiotics - Decreased urine Ca o Thiazides stimulate calcium
• Chlorthalidone • calcium is retained reabsorption in the distal renal
- long half-life (50-60 hrs) due to RBC • calcium is affected by the tubule.
binding parathyroid hormone → • Diabetes insipidus (↓ ADH)
• Indapamide increasing calcium when there is o Nephrogenic diabetes
- with vasodilating activity a drop in calcium levels insipidus (NDI)
- with advantage for HTN - inability to concentrate urine
• Metolazone due to impaired renal tubule
- synergistic with Loop Diuretics response to vasopressin (ADH) =
• Quinethazone Main symptoms are excessive
thirst (polydipsia) and excessive
urine production (polyuria)
o Thiazide diuretics create mild
hypovolemia = ↑salt and water
uptake in proximal tubule =
improved nephrogenic diabetes
insipidus
o ibang mechanism yung involved
sa diuresis na ginagawa ng
Summary: thiazide vs Diabetes Insipidus
- ↑ excretion of: water, sodium, chloride,
potassium and bicarbonate Adverse Effects:
- ↓ excretion of calcium and uric acid - Hypokalemia
• Contraindicated to patients
under digitalis therapy
NOTE: Digoxin toxicity is facilitated by
Hypokalemia.
- Digoxin binds to the K+ site of the
Na+/K+-ATPase pump, low serum
potassium levels increase the risk of
digoxin toxicity.
- Conversely, hyperkalemia diminishes
digoxin's effectiveness. - Low levels of
potassium caused by concurrent digoxin
and diuretics (thiazide & loop diuretics)
may cause weakness, cramps, and
irregular heartbeats.
NOTE: Digoxin toxicity may cause
hyperkalemia.
- Hypercalcemia
- Hyperuricemia
- Hyperglycemia
 - Hyperlipidemia

Drug-Drug Interactions:
• Cholestyramine and Colestipol
(Bile acid sequestrants)
- decrease the absorption of thiazide
diuretics
- Mx: two-hour period allowance
should be observed before the
administration of either drug

NOTE: Hypokalemia = less K+ outside cell (which is blood) = More K+ inside cell
NOTE:
• Digoxin toxicity is facilitated by hypokalemia. Therefore, potassium-wasting diuretics (eg, loop agents, thiazides),
which are often needed in heart failure, can increase the risk of a fatal digitalis arrhythmia.
• Carbonic anhydrase inhibitors, though also potassium-wasting agents, are rarely used for their systemic and
diuretic effects and are therefore less likely to be involved in digitalis toxicity.
• The potassium-sparing diuretics, in contrast to the other groups, can be useful in preventing such interactions
with digitalis but may cause hyperkalemia, which can be arrhythmogenic.

Type Definition/MOA Uses and Adverse Effects

LOOP DIURETICS
- Sulfonamides - aka “high ceiling diuretics” → because more Uses:
• Furosemide (prototype) effective than other diuretics • Drug of choice when rapid and
• Bumetanide - Major site of action: extensive diuresis is needed
• Torsemide • Thick ascending Loop of Henle • Reserve drugs for hypertensive
- Ethacrynic acid - Not a sulfonamide emergency or crisis
but has typical loop activity and some MOA: • Also used in CHF
uricosuric action - inhibit the luminal Na+/K+/2Cl- transporter = • Acute pulmonary edema
inhibit the cotransport of sodium, potassium, and • Acute hypercalcemia (excess
chloride = Marked increase in NaCl excretion calcium in the blood)
• Hyperkalemia
Other effects: • Acute Renal Failure
• Potassium wasting • Anion Overdose (halide
• increased urine Ca and Mg poisoning)
o Bromide and
Fluoride

Adverse Effects:
• Hypokalemic metabolic
alkalosis
o pH of tissues is
elevated

• Spironolactone
> structural analog of
Aldosterone
o Aldosterone
- main mineralocorticoid
hormone (steroid hormone)
- produced by the zona
glomerulosa of the adrenal
cortex of adrenal gland
- essential for sodium
conservation, esp. in the
kidneys
> Active metabolite:
Canrenone
> Absorbed in the GIT and
metabolized in the liver
• Eplerenone
> Spironolactone analog with
greater selectivity for the
aldosterone receptor
• Amiloride and Triamterene
> Onset of action: from 2-4
hours
• Ototoxicity (with amino-
glycosides)
• Hyperuricemia
• Hypomagnesemia
Drug-Drug Interactions:
• Increased risk of ototoxicity
when used with
aminoglycosides and cisplatin
POTASSIUM-SPARING DIURETICS
- They are not potent when used alone, often Spironolactone Uses:
combined with other drugs • Adjunct drug in the mx of
- Major site of action: CHF
• Collecting tubule (after the distal • Conn’s syndrome (adrenal
convoluted tubule, and before the glands produce too much
collecting duct) aldosterone)
Other uses:
MOA: • CHF and edema (combined
- prevent K+ secretion by antagonizing the with thiazide or loop diuretic)
effects of aldosterone at the late distal and • Induces diuresis in patients
cortical collecting tubules with:
- prevent the excretion of potassium o Hyperaldosteronism
- kapag naantagonize ang aldosterone, there will (such as in adrenal
be no sodium retention hyperplasia)
• Spironolactone, Eplerenone o Aldosterone-producing
- direct pharmacologic antagonism of adenomas (benign
mineralocorticoid receptors of tumors in a gland or
aldosterone inhibiting the excretion of gland-like structure)
potassium TRIVIA:
• Amiloride, Triamterene • Hypertrophy = increase in the
- inhibition of Na+ influx through ion size of the cell
channels in the luminal membrane • Hyperplasia = increase in the
number of cells or tissues
Spironolactone Adverse Effects:
• Hyperkalemia
• Menstrual abnormalities in
women
• Gynecomastia, impotence,
and benign prostatic
hyperplasia (BPH)
Spironolactone Contraindications:
• patients with renal
insufficiency (Diabetes
mellitus) and liver disease
Amiloride and Triamterene Uses:
• CHF
• Cirrhosis
• Edema (secondary to
Hyperaldosteronism)
Amiloride and Triamterene Adverse
Effects:
• Hyperkalemia

- osmosis: movement of water from low
concentration of solute to high
concentration of solute
• Mannitol (IV)
• Glycerin (Glycerol)
• Isosorbide
o Mononitrate and
Dinitrate – anti-
anginal drugs
• Urea
- target nila ay enzyme → carbonic
anhydrase
• Acetazolamide
• Methazolamide
• Brinzolamide, Dorzolamide
o topical for glaucoma
o ACE inhibitors can cause
hyperkalemia.
o A drug-drug interaction
o Concomitant use of
Potassium-sparing
diuretics and ACE
inhibitors may lead to
severe hyperkalemia
particularly in patients
with old age, type 2 DM,
and diminished renal
function.
• Nausea and vomiting
• Triamterene: kidney stones
Amiloride and Triamterene
Contraindications:
• Contraindicated in diminished
renal function
OSMOTIC DIURETICS
- They are easily filtered Uses:
- Poorly reabsorbable solutes • To increase urine volume
• To reduce intracranial and
- Site of action: intraocular pressure
• Proximal tubule and descending limb of
Henle's loop Mannitol
• Given intravenously for
MOA: prophylaxis of acute renal
- They pull water into the renal tubule without failure resulting from trauma
sodium loss. or surgery.
- water lang maeexcrete, hindi yung sodium • It increases blood volume and
causes nausea and vomiting.
Mannitol Adverse Effects:
• Dehydration
• Hypernatremia
Isosorbide and Glycerin Uses:
- used for ophthalmic procedures
CARBONIC ANHYDRASE INHIBITORS
- Major site of action: Uses:
• Proximal tubule • Relatively mild diuretic
• Used in glaucoma
MOA: • Used in urinary alkalinization
- Inhibition of the enzyme = prevents • Used in metabolic alkalosis
dehydration of Carbonic acid (H2CO3) and • Prophylaxis in acute mountain
hydration of CO2 in the proximal convoluted sickness
tubule = reduce reabsorption of HCO3 → o Aka altitude sickness
resulting to self-limited diuresis or high-altitude
- when carbonic anhydrase acts on carbonic acid, pulmonary edema
it will be dehydrated into water and CO2 → enter • Adjuvants for the treatment
PCT → hydrated of epilepsy
- carbonic anhydrase inhibitors stop the cycle o Anti-seizure
property
Adverse Effects:
• GI upset
• Urinary infrequency
• Metabolic acidosis
 • Renal calculi formation
• Drowsiness and paresthesias
• Potassium wasting
(hypokalemia)

Drug-drug Interactions:
• Increases the excretion of
salicylates (aspirin) and
lithium (for bipolar disorder)

SODIUM/GLUCOSE COTRANSPORTER (SGLT2) INHIBITORS

• “-gliflozin” – suffix of SGLT2-I - Site of Action: Uses:
• Canagliflozin • Proximal Convoluted Tubule • Diabetes mellitus
• Dapagliflozin o approved for the
• Empagliflozin MOA: treatment of
Inhibition of sodium/glucose cotransporter hyperglycemia, not as
(SGLT2) in the PCT = decreased Na+ and glucose a diuretic
reabsorption = mild diuresis
VASOPRESSIN (ADH) ANTAGONISTS
• “-vaptan” - suffix - Aka Selective Vasopressin V2 Antagonist Tolvaptan Use:
• Tolvaptan - Sometimes called “aquaretics” → water over • For SIADH
• Conivaptan sodium o SIADH = Syndrome of
Inappropriate
- Site of Action: Antidiuretic Hormone
• Collecting Duct secretion → body
makes too much ADH
MOA:
Competitive vasopressin antagonism = Reduces Conivaptan Use:
water reabsorption = increases plasma Na • Has vasodilating effect
concentration (beneficial for hyponatremia) =
increased in urine output

NOTES:
• Demeclocycline (a tetracycline)
o Also arginine vasopressin inhibitor
o Streptococcus aureofaciens
o For therapy of hyponatremia and the syndrome of inappropriate secretion of antidiuretic hormone (SIADH)
• Desmopressin
o A man-made form of vasopressin
o For low level of vasopressin
o Helps to control increased thirst and too much urination due to these conditions, and helps prevent
dehydration.
o Desmopressin is also used to control nighttime bedwetting in children.

OTHER EMERGING GROUPS OF DIURETICS

Na-H EXCHANGER ARGININE VASOPRESSIN XANTHINES
ANTAGONIST RECEPTOR 2 ANTAGONISTS
Dopamine Amphotericin B, Lithium Caffeine, Theophylline (for
DRUG
asthma), Theobromine
Promotes Na+ secretion Inhibits vasopressin’s action Inhibits reabsorption of Na+,
MOA
increase glomerular filtration rate
 SITE OF Proximal Convoluted Tubule Collecting Duct Tubules (esp. proximal tubules)
ACTION

TRIVIA: Caffeine - adenosine antagonist; adenosine is • Adenosine antagonists = induce diuresis and
inhibitory neurotransmitter natriuresis

Adenosine Receptor Antagonist Urearetics

• Rolofylline (KW-3902) is an experimental - These agents are “aquaretics” that increase urea and
diuretic water excretion but not sodium excretion.
o an adenosine receptor antagonist
• Adenosine (also a vasoconstrictor) = lowers
glomerular filtration rate = limits water excretion
 Aspirin, and the remaining group will be the
Negative Control group.
4. Measure the baseline (control) body
temperature by means of a telethermometer
every hour for three hours.
- Cholinergic agonists are substances that “mimic” the 5. Inject subcutaneously (SC) 2 mL of 15%
effect of stimulation of the parasympathetic nervous suspension of Brewer’s Yeast to the test animals
system. with constant temperature.
- Cholinergic antagonists are drugs that oppose or 6. Measure the temperature of each animal after
nullify the effect of stimulation of the parasympathetic eighteen to twenty-four hours.
nervous system. 7. Give orally to the animals the computed dose of
Positive controls per kg body weight (BW).
Test Animal (Note: Control is normal saline solution.)
• Rabbits 8. Measure every hour for three hours the body
temperature of each animal.
Drugs 9. Note and record the results.
• Atropine SO4 - 1 gtt of a 0.5 to 1% solution
o Antimuscarinic cholinolytic agent 3 Animal Groups
o Causes mydriasis (pupil dilation) - Negative Control Group
• Pilocarpine HCl – 1 gtt of a 0.5 to 1% solution • Normal saline solution/Distilled Water - Rabbit
o Alkaloid cholinomimetic agent #1
o Causes Miosis (pupil constriction) • Normal saline solution/Distilled Water - Rabbit
#2
o NOTE: Ideal Volume Limit for Oral
Gavage (Rabbits) = 10 mL/kg BW
Materials: o NOTE: Ideal Volume Limit for
• Alcohol Normal Subcutaneous (Rabbits) = 1-5 mL/kg BW
• Saline Solution
- Positive Control Groups
• Brewer’s Yeast
• Paracetamol - Rabbit #3 (100 mg/kg PO)
• Plastic container
• Aspirin - Rabbit #4 (10 mg/kg PO)
• Cage
• Syringe
Test Animal Group Weight (g)
• Gloves
1 2,128 g
• Telethermometer (-) control
2 1,492 g
(+) Control
Test Animal 3 1,250 g
Paracetamol
• Rabbits (+) Control
4 1,488 g
Aspirin
Drugs
• Paracetamol = 100-500 mg/kg (varies Computation for NSS/Distilled Water
depending on references Dose Computation
• Aspirin = 10-100 mg/kg (varies depending on • Negative Control groups:
references o Rabbit #1
▪ Ideal Volume Limit for Oral
Procedures Gavage = 10 mL/kg BW
1. Prepare all the materials needed. ▪ Weight = 2,128 g = 2.128 kg
2. Weigh the animals to be treated and divide o Rabbit #2
them into three groups. ▪ Ideal Volume Limit for Oral
3. Mark to identify them. One group will be treated Gavage = 10 mL/kg BW
with Paracetamol, one group will be given ▪ Weight = 1,492 g = 1.492 kg
Volume Limit (Using the standard 10 mL/kg BW) Volume Limit (Using the standard 10 mL/kg BW)
• Rabbit #1 = 2,128 g = 2.128 kg • Rabbit #3 = 1,250 g = 1.25 kg
10 𝑚𝐿 𝑥 10 𝑚𝐿 𝑥
= =
1 𝑘𝑔 2.128 𝑘𝑔 1 𝑘𝑔 1.25 𝑘𝑔
x = 21.28 mL volume of NSS/DW to be orally given to x = 12.5 mL volume of drug to be given orally to the 1.25
the 2.128 kg rabbit kg rabbit

• Rabbit #2 = 1,492 g = 1.492 kg Dose (mg) for Rabbit #3 Paracetamol (100mg/kg PO)
10 𝑚𝐿 𝑥 100 𝑚𝑔 𝑥
= =
1 𝑘𝑔 1.492 𝑘𝑔 𝑘𝑔 1.25 𝑘𝑔
x = 14.92 mL volume of NSS/DW to be orally given to x = 125 mg of drug to be given orally to the 1.25 kg
the 1.492 kg rabbit rabbit

Dose (mg) for NSS or DW Volume of 15% Brewer’s Yeast Suspension

• NOT APPLICABLE - Using 2 mL/kg, the volume of 15% Brewer's yeast
o NSS/DW is only a vehicle! suspension to be administered subcutaneously is:
o No relevant therapeutic effect to the • Rabbit #3 = 1,250 g = 1.25 kg
study! 2 𝑚𝐿
=
𝑥
o You don't need to compute using 0.9% 𝑘𝑔 1.25 𝑘𝑔
concentration because you will use NSS x = 2.5 mL of 15% Brewer's yeast suspension SC for 1.25
as is! (No mixing or compounding kg rabbit
involved!)
Computation for Aspirin
Computation for the Volume of 15% Brewer’s Yeast • Positive Control Group
Suspension o Aspirin (10mg/kg PO)
- Using 2 mL/kg, the volume of 15% Brewer's yeast ▪ Ideal Volume Limit for Oral
suspension to be administered subcutaneously is: Gavage = 10 mL/kg BW
• Rabbit #1 = 2,128 g = 2.128 kg ▪ Weight of Rabbit #4 = 1,488 g =
2 𝑚𝐿 𝑥 1.488 kg
=
𝑘𝑔 2.128 𝑘𝑔
x = 4.256 mL of 15% Brewer's yeast suspension SC for Volume Limit (Using the standard 10 mL/kg BW)
2.128 kg rabbit • Weight of Rabbit #4 = 1,488 g = 1.488 kg
10 𝑚𝐿 𝑥
=
NOTE: Although the standard volume limit for 1 𝑘𝑔 1.488 𝑘𝑔
subcutaneous (SC) is 1-5 mL/kg BW for rabbit, use the x = 14.88 mL volume of drug to be given orally to the
specific volume limit for 15% Brewer's yeast suspension 1.488 kg rabbit
that is provided protocol in the experiment.
Dose (mg) for Rabbit #4 Aspirin (10 mg/kg PO)
10 𝑚𝑔 𝑥
• Rabbit #2 = 1,492 g = 1.492 kg =
2 𝑚𝐿 𝑥 𝑘𝑔 1.488 𝑘𝑔
= x = 14.88 mg of drug to be given orally to the 1.488 kg
𝑘𝑔 1.492 𝑘𝑔
x = 2.984 mL of 15% Brewer's yeast suspension SC for rabbit
1.492 kg rabbit
Volume of 15% Brewer’s Yeast Suspension
Computation for Paracetamol - Using 2 mL/kg, the volume of 15% Brewer's yeast
• Positive Control Group suspension to be administered subcutaneously is:
o Paracetamol (100mg/kg PO) • Rabbit #4 = 1,488 g = 1.488 kg
2 𝑚𝐿 𝑥
▪ Ideal Volume Limit for Oral =
𝑘𝑔 1.488 𝑘𝑔
Gavage = 10 mL/kg BW
x = 2.976 mL of 15% Brewer's yeast suspension SC for
▪ Weight of Rabbit #3 = 1,250 g =
1.488 kg rabbit
1.25 kg
% Protection/Reduction Rabbit #2: NSS/DW
𝑩 − 𝑪𝒏 38.7 − 39.47
( ) × 𝟏𝟎𝟎% ( ) × 100 = −48.125%
𝑩−𝑨 38.7 − 37.1
• B = temperature after pyrexia induction (after NO REDUCTION IN TEMPERATURE, RATHER INCREASE.
Brewer's yeast)
• Cn = post-treatment temperature (average) Rabbit #3: Paracetamol
39 − 37.97
o Get the average (1-3 hr) ( ) × 100 = 60.59%
39 − 37.3
• A = normal body temperature (baseline) REDUCTION IN TEMPERATURE

% Reduction Computation for NSS / DW Rabbit #4: Aspirin

Rabbit #1: NSS/DW 39.5 − 38.33
38.1 − 39.1 ( ) × 100 = 68.82%
( ) × 100 = −83.33% 39.5 − 37.8
38.1 − 36.9 REDUCTION IN TEMPERATURE
NO REDUCTION IN TEMPERATURE, RATHER INCREASE.

Test Vol.
Weight Dose %
Animal Group admin. Temperature (Deg. Celsius)
(g) (mg) Protection
No. (mL)
After Ave.
Baseline Brewer’s 1st hr 2nd hr 3rd hr Temp.
Yeast (1-3 hrs)
1 2,128 X 21.28 36.9 38.1 38.7 39.1 39.5 39.1 -83.33
(-) control
2 1,492 X 14.92 37.1 38.7 38.9 39.5 40 39.47 -48.125
(+) control
3 1,250 125 12.5 37.3 39 38.7 38.1 37.1 37.97 60.59
Paracetamol
(+) control
4 1,488 14.88 14.88 37.8 39.5 39.2 38.5 37.3 38.33 68.82
Aspirin

Conclusion targeting cyclooxygenase-1 (COX-1) and

- Aspirin and Paracetamol, as antipyretics, reduced body cyclooxygenase-2 (COX-2).
temperature induced by Brewer’s yeast in rabbits. • Paracetamol
o weak inhibitor of the synthesis of
Use of Brewer’s yeast in the Experiment prostaglandins (PGs) by reducing the
• To induce pyrexia/fever active form of COX-1 and COX-2
• In theory... enzymes.
o After 18 hr of Brewer’s yeast injection,
there is rise in temperature of at least
0.6°C (or 1°F).

Telethermometer
• To determine temperatures at a distance by an
electric current
o Inserted in the rabbit’s rectum
o Commonly used in pyrogen testing
▪ Pyrogen – substances that
induce fever

Aspirin and Paracetamol MOA

• Aspirin
o NSAID
o disrupts the production of
prostaglandins throughout the body by