personal lab report

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INDEX
Serial No. Name of Experiment Date Page
No.
01. Neuromuscular Strength Induced in
mice (Swiss Albino)
06/02/2019 02-07

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Experiment no: 01
Date: 06/02/2019
Name of experiment: Neuromuscular Strength Induced in mice (Swiss Albino)
Background of Drug:
Description:
o Diazepam is a benzodiazepine tranquilizer with anticonvulsant, sedative,
muscle relaxant and amnesic properties.
o It is used in the treatment of anxiety and tension states and also as premedicant
in the control of muscle spasm.
o Diazepam induces calming effect by acting on parts of the limbic system, the
thalamus and hypothalamus.
o Diazepam is one of the most rapidly and completely absorbed benzodiazepines,
reaching peak concentrations in about an hour in adults, but as quickly as 15 to
30 minutes in children.
o Secondary peaks in the plasma concentration have been described for diazepam
at 6 to 12 hours after an oral dose. This is most likely due to enterohepatic
recirculation.
o Diazepam is bound to plasma protein to a great extent (98.5%).
o After intravenous administration of diazepam, it is redistributed in a manner
typical of that for highly lipid soluble agents.
o Central effects develop promptly but wane rapidly as the drugs move to other
tissue. Diazepam is extensively metabolized in the liver.
o It is excreted in the urine, mainly in the form of its metabolites, either free or in
conjugated form. There is no biliary excretion.

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Dose:
Indication:
Diazepam has a number of uses including:
 Treatment of anxiety, panic attacks, and states of agitation
 Treatment of neurovegetative symptoms associated with vertigo
 Treatment of the symptoms of alcohol, opiate, and benzodiazepine withdrawal
 Short-term treatment of insomnia
 Treatment of muscle spasms
 Treatment of tetanus, together with other measures of intensive treatment
 Adjunctive treatment of spastic muscular paresis (paraplegia/tetraplegia) caused by
cerebral or spinal cord conditions such as stroke, multiple sclerosis, or spinal cord injury
(long-term treatment is coupled with other rehabilitative measures)
 Palliative treatment of stiff person syndrome
 Pre- or postoperative sedation, anxiolysis or amnesia (e.g., before endoscopic or surgical
procedures)
 Treatment of complications with a hallucinogen crisis and stimulant overdoses and
psychosis, such as LSD, cocaine, or methamphetamine
 Preventative treatment of oxygen toxicity during hyperbaric oxygen therapy

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Contraindication and precaution:
Use of diazepam should be avoided, when possible, in individuals with:
 Ataxia
 Severe hypoventilation
 Acute narrow-angle glaucoma
 Severe hepatic deficiencies (hepatitis and liver cirrhosis decrease elimination by a factor
of two)
 Severe renal deficiencies (for example, patients on dialysis)
 Liver disorders
 Severe sleep apnea
 Severe depression, particularly when accompanied by suicidal tendencies
 Psychosis
 Pregnancy or breast feeding
 Caution required in elderly or debilitated patients
 Coma or shock
 Abrupt discontinuation of therapy
 Acute intoxication with alcohol, narcotics, or other psychoactive substances (with the
exception of some hallucinogens or stimulants, where it is occasionally used as a
treatment for overdose)
 History of alcohol or drug dependence
 Myasthenia gravis, an autoimmune disorder causing marked fatigability
 Hypersensitivity or allergy to any drug in the benzodiazepine class
Side effect:
The side effects of Diazepam are infrequent and mild.
 Drowsiness,
 Dry Mouth,
 Inattentiveness May Occur And Are Dose Dependent.
 Light Headedness,
 Slight Ataxia,
 Vertigo,
Other effects that may be observed rarely include
 Gastro-Intestinal And
 Hypotension,
 Visual Disturbances.
In case of long term medication change in libido may occur.

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Drug interaction:
If diazepam is given concomitantly with centrally acting drugs such as neuroleptics,
tranquillisers, antidepressants, hypnotics, analgesics and anaesthetics, the sedative effects are
likely to be intensified.
Known inhibitors of hepatic enzymes
E.g. cimetidine and omeprazole have been shown to reduce the clearance of diazepam
and may potentiate their action and known inducers of hepatic enzyme,
E.g. rifampicin may increase the clearance of diazepam.
Use in pregnancy and lactation:
The use of Diazepam during the first trimester of pregnancy should almost always be
avoided as it bears a risk of congenital malformation.
Diazepam has been detected in breast milk.
If possible the use of diazepam should be avoided during lactation.
Neuromuscular Strength:
Strength Training and Neuromuscular Adaptations. Systematic strength training
produces structural and functional changes, or adaptations, in the body. The level of adaptation
is evidenced by the size and strength of the muscles.
These drugs fall into two groups:
 Non-depolarizing blocking agents:
These agents constitute the majority of the clinically relevant neuromuscular blockers.
They act by competitively blocking the binding of ACh to its receptors, and in some
cases, they also directly block the ionotropic activity of the ACh receptors.
 Depolarizing blocking agents: These agents act by depolarizing the sarcolemma of the
skeletal muscle fiber. This persistent depolarization makes the muscle fiber resistant to
further stimulation by ACh.
Study design:
 Collection of experimental animals: Swiss albino mice of either sex, 6-7 weeks, weighing
25–30 gm.
 Storage: The mice were stored in well ventilated cages in animal house of pharmacy
department, Jashore University of science and technology.
Non-depolarizing blocking agents Depolarizing blocking agents
Tubocurarine Succinylcholine
Mivacurarium Diazepam
Cisatracurium

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 Temperature and Humidity: Mice were kept under standard environmental conditions,
having relative humidity 55%–65%, and (27.0 ± 1.0) ∘C temperature.
 Light cycle: 12 h light/12 h dark cycle.
 Food and water: Proper formulated rodent pellet foods and water ad libitum were ensured.
Before the experiment.
 Adaptation: Animals were adapted to the laboratory conditions for 1 week.
1.1 Apparatus :
Kondziela's inverted screen test
The inverted screen is a 43 cm square of
wire mesh consisting of 12 mm squares of
1 mm diameter wire (Figure 1). It is
surrounded by a 4 cm deep wooden
beading (which prevents the occasional
mouse which attempts to from climbing on
to the other side)
Figure 2: inverted screen test
1.2 Weights test
Five weights constitute the apparatus. Each
consists of a ball of tangled fine gauge
stainless steel wire, a "scale collector" as used
to prevent limestone scale formation in
domestic kettles. The weight of this is small (7
g) but each scale collector is attached to a
series of steel chain links, each weighing
approximately 13 g. The number of links
ranges from one to seven. Their weights are
therefore: 20, 33, 46, 59, 72, 85 and 98 g.
Figure: Close view of a mice on the inverted screen
Procedure
For all tests, bring the mice to the experimental room 5-20 min before testing to ensure they
are properly awake. As a general rule, to allow recovery of muscular strength and a return to
normal levels of arousal, rest the mice by a return to the home cage after each motor test.

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Kondziela's inverted screen test
Procedure: Place the mouse in the centre of the wire mesh screen, start a stop clock, rotate
the screen to an inverted position over 2 sec, with the mouse's head declining first. Hold
the screen steadily 40-50 cm above a padded surface. Note the time when the mouse falls
off, or remove it when the criterion time of 60 sec is reached. Longer criterion times may
be useful for some experiments.
Control group:
Mice no. 1st Test Time(Sec) 2nd Test Time(Sec) 3rd Test Time(Sec) Average Time(Sec)
1. 10 8 11 9.6
2. 15 8 12 11.6
3. 9 12 7 9.3
4. 10 6 9 8.3
5. 8 10 15 11
Standard group:
Comments:
Above these average result between Standard group vs Control group, the control group is
more significant than standard group for neuromuscular strength determining.
Precaution:
 Room temperature ,sound air must be comfortable for mice
 Weight of mice must be 20 - 98 gm
 Dose will be small amount because of low wt. of mice.
Mice no. 1st Test Time(Sec) 2nd Test Time(Sec) 3rd Test Time(Sec) Average Time(Sec)
1. 8 5 6 6.33
2. 12 4 10 8.66
3. 6 10 4 6.66
4. 9 4 4 5.66
5. 6 14 8 9.33

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Md. Jahidul Islam
Session: 2015-16
Department pharmacy
Jashore University of science and technology