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aubf lecture final term > CSF > Flashcards

CSF Flashcards

1
Q

It is the major fluid in the body that surrounds the brain and spinal cord.

A

CEREBROSPINAL FLUID

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2
Q

Main function of CSF

A

protect brain and spinal cord by
acting as a lubricant, cushion for transport of nutrients, and
metabolic waste management.

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3
Q

Three main reasons of why we perform CSF in lab:

A
  1. To evaluate the function of central nervous system
  2. It indicates presence of infections, autoimmune disorders,
    diseases of the brain and spinal cord
  3. It can also detect early signs of seizures and dementia
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4
Q

is the infection of the membrane
surrounding our brain and spinal cord. E.g.,
bacterial, fungal, and viral meningitis.

A

Meningitis

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5
Q

Two main sources of CSF:

A

Choroid plexus (70%)
Subarachnoid space (30%)

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6
Q

lines the choroid plexus which is responsible for the formation of blood-brain barrier.

A

Choroidal cells (epithelial cells)

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7
Q

regulates the movement of ions, molecules, and cells between the blood and brain.

A

Blood brain barrier

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8
Q

Three Layers of meninges that protect the brain and spinal cord

A

Dura – outermost “hard mother”
Arachnoid – middle “spiderweb-like”
Pia – innermost “gentle mother

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9
Q

Three Types of Epithelial Cells

A

Ependymal cells
Choroidal cells
Pia Arachnoid Mesothelial cells (PAM)

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10
Q

lines the cerebral ventricles and pleural canals of the spinal cord. Range in shape from squamous to columnar. May be ciliated which line the central cavities of the brain and spinal column. Separate the CNS interstitial fluid from the cerebrospinal fluid in the
cavities.

A

Ependymal cells

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11
Q

main source of CSF formation

A

Choroidal cells

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12
Q

lines the mesoderm of pia and arachnoid mater.

A

Pia Arachnoid Mesothelial cells (PAM)

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13
Q

CSF ionic components such as H, K, Ca, bicarbonates, and Mg are tightly regulated by a?

A

specific transport system.

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14
Q

T or F
Glucose, urea, and creatinine diffuse freely and require 2 or more hours to regulate.

A

T

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15
Q

Volume of CSF each day

A

500 mL of CSF

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16
Q

Range of 500 mL CSF/day

A

0.3 to 0.4 mL/min.

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17
Q

Range of Adult volume of CSF

A

90 –150 mL

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18
Q

? mL in the ventricles and subarachnoid space (adult volume range)

A

25 mL

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19
Q

Neonates volume range of CSF

A

10 – 60 mL

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20
Q

Total CSF volume is replaced every - hours because of proper metabolic waste management of our body.

A

every 5-7 hours

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21
Q

The process or the procedure for obtaining CSF fluid is known as

A

cisternal or lumbar puncture or spinal tap (through ventricular cannulas or shunts)

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22
Q

The most common site for lumbar puncture

A

intervertebral space

between your L3 and L4 for adults.
But for pediatric patients L4 and L5.

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23
Q

T or F
only doctors can perform lumbar puncture procedure.

A

T

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24
Q

Normal Opening pressure for Adults

A

90 – 180 mm of water

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25
Q

Normal Opening pressure for Obese Patients

A

250 mm of water

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26
Q

Normal Opening pressure for Infants and children

A

10 – 100 mm of water attaining
adult range by 6-8 years of age.

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27
Q

if there is >250 mm of water

A

increased intracranial pressure which imply presence of
intracranial hemorrhage, and tumors.

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28
Q

if there is >200 mm of water

A

in a relaxed patient, no more than 2.0 mL should
be withdrawn.

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29
Q

When collecting CSF, we are using?

A

manometer

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30
Q

indicates the pressure because if too much pressure, it can
indicate obesity, intracranial pressure will also imply presence of bacterial infection, meningitis, hemorrhage, and tumors.

A

Manometer

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31
Q

Define VDRL and Identify what type of test do they use?

A

Venereal Disease Research Laboratory, test for syphilis

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32
Q

India ink is used for?

A

Cryptococcus specimens

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33
Q

is the most sterile among the three csf tubes.

A

Tube 2

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34
Q

Contaminants in tube 2 should be avoided because if there is, it can indicate _____?

A

false positive results

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35
Q

GROSS EXAMINATION of CSF
Normal?

A

crystal clear or colorless

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36
Q

GROSS EXAMINATION
Viscosity?

A

watery

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37
Q

Present of Turbidity and Cloudiness, basaha lang

A

o White blood cells > 200 uL
o Red blood cells > 400 uL
o Microorganisms
o Radiographic contrast media
o Aspirated contrast media
o Aspirated epidural fat
o Proteins > 150 mg/dL (1.5 g/L)

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38
Q

Bloody/pink Turbidity may indicate?

A

RBC > 6000 uL
- Subarachnoid and intracerebral hemorrhage
- Cerebral infarct or traumatic spinal tap
- It can be traumatic or hemorrhagic

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39
Q

If hemorrhagic, 3 tubes will turn into what color?

A

all (3) tubes color is red.

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40
Q

If improperly collected traumatic tap, 3 tubes will turn into what color?

A

first tube red, second tube light red, and third tube clear/not equal.

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41
Q

Refers to presence of oxidized hemoglobin from lysed red blood cells.

A

Xanthochromia and Associated Disease

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42
Q

Pink CSF may indicate?

A

Red blood cells lysis

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43
Q

Orange CSF may indicate?

A

-Red blood cells lysis | hemoglobin breakdown
- 2-4 hours after subarachnoid hemorrhage and take as
long as 12 hours, peak 24-36 hours and disappears on 4-
8 days.
- Hypervitaminosis A (carotenoids)

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44
Q

Yellow CSF may indicate?

A

-RBC lysis | hemoglobin breakdown
- Hyperbilirubinemia: develops after 12 hours after
subarachnoid bleed, peaks 2-4 days and persists for 2-4
weeks.
- CSF protein > 150 mg/dL (1.5 g/L)

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45
Q

Yellow green CSF may indicate?

A

Hyperbilirubinemia (biliverdin)

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46
Q

Brown CSF may indicate?

A

Meningeal metastatic melanoma

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47
Q

(3) Clot Formation

A

o Traumatic taps
o Complete spinal block (Froin’s syndrome)
o Suppurative or tuberculous meningitis

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48
Q

(3) Presence of High Viscosity

A

o Metastatic mucin producing adenocarcinomas
o Cryptococcal meningitis
o Liquid nucleus pulposus

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49
Q

You need to test these 4 parameters to differentiate the type of meningitis present.

A

PROTEIN
GLUCOSE
WBC POPULATION
LACTATE

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50
Q

MICROSCOPIC EXAMINATION
Identify and give its CV%.
18 large squares (1 mm2 each) depth of 0.2 m
A total volume of 3.6 uL (18 x 0.2) is examined

A

Fusch-Rosenthal Chamber
CV = 48%

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51
Q

MICROSCOPIC EXAMINATION
Identify and give its CV%.
9 1mm2 square with a depth of 0.1 mm

A

Neubauer Hemocytometer
CV = 45%

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52
Q

An automated type of cytometry that Yield rapid and reliable WBC and RBC counts

A

UF – 100 Flow Cytometer

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53
Q

in UF – 100 Flow Cytometer, what tube is utilized for the cell count

A

tube 3

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54
Q

T or F
In doing cell counting of CSF, you need to do it immediately since your white blood cells may lyse.

A

T

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55
Q

T or F
Proper storage of CSF just at 37C.

A

F
just at room temperature. It should not be refrigerated because a lot of elements can be present during refrigeration.

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56
Q

What Department of the laboratory?

common fastidious organisms such as your Hemophilus and your Neisseria which are the main contributors during an infection in CSF specimens cannot live in cold temperatures.

A

Microbiology Department

57
Q

T or F
Remain undiluted if the CSF is clear.

A

T

58
Q

(2) If the CSF is bloody, dilute it with?

A

normal saline solution (nss)
glacial acetic acid

59
Q

for white blood cells to be clearer.

A

Crystal violets

60
Q

The dilution of CSF

A

1:10 or 1:20 dilution if the specimen is cloudy.

61
Q

remember (basaha lang)

A

o Charge for the cells to settle.
o Large squares constant measurement 0.1
o Small squares constant measurement 0.004

62
Q

Normal Leukocyte Count for Adults

A

0 -5 cells /uL

63
Q

Normal Leukocyte Count for Neonates

A

0 – 30 cells/uL

64
Q

No RBC should be present in CSF (take note) . Unless if present, it is a sign of???

A

hemorrhage

65
Q

Correcting leukocytes and protein introduced by?

A

traumatic tap

66
Q

refers to the leukocytes added to CSF by traumatic tap. Generally, this formula can be applicable during a traumatic tap.

A

WBC added

67
Q

T or F
Differential Counting of CSF specimens is advised

A

F

68
Q

(2) Direct smears of the centrifuged CSF sediment are also subject to significant error from?

A

cellular distortion and fragmentation

69
Q

The stain used for CSF is the

A

Wright’s Stain

70
Q

Recommended method for differential count of all body
fluids.

A

CYTOCENTRIFUGE

71
Q

-Rapid, requires minimal training and allows Wright’s staining of air-dried cytospins.
-Cell yield preservation are better than with simple
centrifugation.

A

CYTOCENTRIFUGE

72
Q

CYTOCENTRIFUGE
___-___ cells can be concentrated from ____ mL of normal
CSF.

A

30 – 50 cells can be concentrated from 0.5 mL of normal
CSF.

73
Q

Familiarize this

A

Variable artifactual distortions may be seen but can be
minimized when the specimen is fresh, albumin is added to
the specimen (2 drops of bovine serum albumin) and cell
concentration is adjust to about 300 WBC/L prior to
centrifugation.

74
Q

Too cumbersome

A

FILTRATION AND SEDIMENTATION

75
Q

Adults:
Lymphocytes and monocytes Ratio

A

70:30 ratio (abundant)

76
Q

Adults:
Neutrophils

A

2±5

77
Q

Adults:
Histiocytes, ependymal cells, and eosinophils

A

RARE

78
Q

Young children (basaha na largo uy)

A
  • Monocytes – 80% (abundant)
  • Lymphocytes – 20±18
  • Neutrophils - 3±5
  • Histiocytes - 5±4
  • Ependymal cells and eosinophils – rare
79
Q

(3) CSF Examination for tumor cells has the sensitivity to:

A

o Leukemic patients (70%) – all
o Metastatic carcinoma (20-60%)
o Primary CNS malignancies (30%)

80
Q

What is present?
It has 4-5 lobes, segmented, granules are present. So, this is a sign of bacterial infection.

A

Presence of neutrophils

81
Q

Causes of INCREASED CSF NEUTROPHILS

A
  1. Meningitis
  2. Other infections:
    o Cerebral abscess
    o Subdural empyema
    o AIDS – related CMV radiculopathy
  3. Following seizures
  4. Following CNS hemorrhage
  5. Following CNS infarct
  6. Reactions to repeated lumbar punctures
  7. Injection of foreign material (methotrexate; contrast media)
  8. Metastatic tumor in contact with CSF

Presence of lymphocyte and monocyte (please memorize this)

82
Q

Causes of CSF LYMPHOCYTOSIS

A

Meningitis
Degenerative (kamo nay basa)
Other inflammatory disorders (kamo nay basa)

83
Q

cytoplasm and clump chromatins are signs of these WBCs. This can be seen during multiple sclerosis or other degenerative disorders.

A

Reactive Lymphocytes

84
Q

Causes of CSF PLASMACYTOSIS

A
  1. Acute viral infections
  2. Guillain-Barre syndrome
  3. Multiple sclerosis
  4. Parasitic CNS infections
  5. Sarcoidosis
  6. SSPE
  7. Syphilitic meningoencephalitis
  8. Tuberculous meningitis

Presence of eosinophils (please memorize this)

85
Q

Causes of EOSINOPHILIC PLEOCYTOSIS

A
  1. Acute polyneuritis
  2. CNS reactions to foreign material (drugs, shunts)
  3. Fungal infections
  4. Idiopathic hypereosinophilic syndrome
  5. Parasitic infections

Presence of macrophage and Presence of macrophage with hemosiderin granules (please memorize this)
-Macrophages can indicate previous hemorrhage of your CSF.

86
Q

(2) Nonpathologically Significant Cells

A

Clustered ependymal cells
Choroid plexus cells in CSF

87
Q

Cells that line the cerebral ventricles and pleural canal of
your spinal cord.

A

Clustered ependymal cells

88
Q

These cells form your blood-brain barrier. This is the main
source of the CSF formation.

A

Choroid plexus cells in CSF

89
Q

(2) Malignant Cells

A

Blast cell in CSF
Medulloblastoma

90
Q

Commonly found in leukemic patients. Pathologic origin
(lymphoblast, myeloblast, monoblast). Nonpathological
origin include astrocytoma, retinoblastoma, medulloblastoma

A

Blast cell in CSF

91
Q

A malignant cell which can indicate metastatic cancers.

A

Medulloblastoma

92
Q

CHEMICAL ANALYSIS
Proteins (basaha lang)

A

-Increased presence of proteins can be seen in
pathologic conditions.
-Decreased proteins can be found in fluid-leakage in CNS
-1% of the plasma level

93
Q

Normal level of protein in adults:

A

15 – 45 mg/dL

94
Q

(#) CSF protein fell rapidly from birth to 6 months of age

A

(118 - 40 mg/dL), plateaued 3 -10 years (32 mg/dL) and
rose slightly from 10 -16 years of age (41 mg/dL)

95
Q

Classic Lowry method - __-__mg/dL

A

24.1 – 48 mg/dL

96
Q

Trichloroacetic acid-ponceau S method - __-__mg/dL

A

15 – 49 mg/dL

97
Q

Biuret method - __-__mg/dL

A

22.3 – 50.3 mg/dL

98
Q

3 Types of protein that can be seen in the chemical analysis of CSF

A

Albumin
Pre-albumin (transthyretin)
Transferrin (beta 2 transferrin or tau)

99
Q

what protein is high because of its dual synthesis by the liver and choroid plexus.

A

Pre-albumin (transthyretin)

100
Q

what protein is present in electrophoresis, it
migrates slowly than in serum owing to cerebral
neuraminidase digestion of sialic acid residues.

A

Transferrin (beta 2 transferrin or tau).

101
Q

2 Methodology of Proteins

A

Turbidimetric methods
Colorimetric methods (protein determination)

102
Q

(4)Turbidimetric methods

A

a. Trichloroacetic acid (TCA)
b. Sulfosalicylic acid (SSA)
c. Sodium sulfate for protein precipitation
d. Benzethonium chloride or Benzalkonium chloride

103
Q

Pros and cons of Turbidimetric methods

A

Advantages: popular, simple, rapid, and require no special
instrumentation.

Disadvantages: temperature sensitive and require large volumes (0.5 mL)

False protein elevation: using TCA methods in the presence of methotrexate

104
Q

(2) Colorimetric methods (protein determination)

A

a. Lowry method
b. Dye binding methods
c. Modified Biuret method

105
Q

the stain used in Dye binding methods of colorimetric determination of protein

A

Coomassie Brilliant Blue (CBB), a rapid, highly sensitive,
uses small samples (25 – 50 u/L).

106
Q

(4) Electrophoretic Techniques

A
  1. CSF electrophoresis
  2. Electroimmunodiffusion
  3. Immunofixation electrophoresis
  4. Ig Immunoblotting
107
Q

CSF electrophoresis characteristics, Basaha lang

A
  • MS: gammaglobulin fraction
  • high resolution agarose gel electrophoresis
  • MS: discrete populations of IgG
  • the oligoclonal bands (kappa and lambda)
  • Coomassie brilliant blue (CBB) or paragon violet stains resolve oligoclonal bands in only 5 ug of IgG
  • Silver staining – 20-50x sensitive than CBB and be used on unconcentrated CSF
  • Isoelectric focusing
  • Polyacrylamide gel
  • MS – CSF IgG | albumin ratio is > 0.25
108
Q

Identify what type of Electrophoretic Techniques
* More sensitive than agarose gel electrophoresis
* Does not require CSF concentration
* Provide fewer oligoclonal bands; more diffuse bands

A

Immunofixation electrophoresis

109
Q

(8) Other CSF Proteins

A

Myelin Basic Protein (MBP)
Alpha-2-macroglobulin (A2M)
Beta-2-microglobulin (B2M)
C-Reactive Protein (CRP)
Fibronectin
Beta-Amyloid Protein 42 and Beta 2 Transferrin (Tau)
Protein 14-3-3
Transferrin

110
Q

Identify what type in Other CSF Proteins
- Seen in MS
- Correlate with CSF leukocyte count, intrathecal IgG synthesis, and CSF | serum albumin concentration quotient.
- Surrogate marker during acute MS exacerbations

A

Myelin Basic Protein (MBP)

111
Q

Identify what type in Other CSF Proteins
Increased in pinocytic vesicles which reflects hemorrhage or breakdown of BBB, as occurs in bacterial meningitis.

A

Alpha-2-macroglobulin (A2M)

112
Q

Identify what type in Other CSF Proteins
- Part of HLA class I molecule
- >1.8 mg/L = leptomeningeal leukemia or lymphoma
- Marker of neuro-Bechet’s syndrome
- HIV-1 and various malignancies have been associated

A

Beta-2-microglobulin (B2M)

113
Q

Identify what type in Other CSF Proteins
- Acute phase reactant marker can indicate
bacterial meningitis esp. during large amounts.
- Differentiating viral (aseptic) meningitis from
bacterial meningitis (increased)

A

C-Reactive Protein (CRP)

114
Q

Identify what type in Other CSF Proteins
Cell adhesion allows leukocytes to adhere and pass
through the vascular endothelia and migrate to the
inflammatory site

A

Fibronectin

115
Q

Identify what type in Other CSF Proteins
Increased CSF levels of microtubule-associated tau protein
and decreased beta-amyloid protein 42 and presence of neurofibrillary tangles and amyloid plaques = diagnosis of Alzheimer’s disease (AD)

A

Beta-Amyloid Protein 42 and Beta 2 Transferrin (Tau)

116
Q

Identify what type in Other CSF Proteins
- Present in Creutzfeldt-Jacob Disease (CJ)
- 2 proteins (130 and 131) have the same amino acid
sequence as protein 14-3-3
- Determined by immunoassay
- False (+) = stroke and meningoencephalitis

A

Protein 14-3-3

117
Q

Identify what type in Other CSF Proteins
- CSF leakage following head trauma and recurrent
meningitis makes accurate diagnosis of the leaking
fluid.
- Immunofixation electrophoresis identifies both
- Protein electrophoresis is non-invasive, rapid,
inexpensive, and requires 0.1 mL

A

Transferrin

118
Q

(4) Increased Fibronectin indicates?

A

a. All = poor prognosis
b. Burkitt’s lymphoma
c. Bacterial meningitis
d. Astrocytomas

119
Q

(2) decreased Fibronectin indicates?

A

a. Viral meningitis
b. AIDS dementia complex

120
Q

two types of Transferrin?

A

a. Beta-1-transferrin – present in body fluids
b. Beta-2-transferin – present only in the CNS;

121
Q

What type of transferrin that allows the conversion from Beta-1-transferrin by neuraminidase?

A

Beta-2-transferin

122
Q

Glucose basaha lang

A
  • Fasting CSF glucose: 50 – 80 mg/dL (2.8 – 4.4 mmol/L)
  • CSF glucose comprises 60% of plasma values
  • Normal fasting hours is 8 hrs. and results should be compared with plasma following 4 hours fast
  • Normal CSF/plasma glucose ratio: 0.3 – 0.9 with
    fluctuations of blood levels because of lag in CSF glucose
    equilibration time.
  • decreased 40 mg/dL (2.2 mmol/L) or ratio < 0.3 hypoglycorrhachia
  • Increase in presence of glucose or large amounts can
    indicate Bacterial, tuberculous, fungal meningitis.
123
Q

Lactate basaha lang more on ranges

A
  • Children and adults: 9.0 – 26 mg/dL (1.0 – 2.9 mmol/L)
  • Newborns first 2 days of life: 10 – 60 mg/dL
  • Newborns 3 – 10 days old: 10 – 40 mg/dL (1.1 – 4.4 mmol/L)
  • Elevated levels (increased) reflect CNS anaerobic metabolism due to tissue hypoxia.
  • Viral meningitis: < 25 mg/dL
  • Bacterial meningitis: > 35 mg/dL = cut off value 30-36
124
Q

Lactate range for adults mg/dL

A

9.0 – 26 mg/dL (1.0 – 2.9 mmol/L)

125
Q

Lactate range for Newborns first 2 days of life mg/dL

A

10 – 60 mg/dL

126
Q

Lactate range for Newborns 3 – 10 days old mg/dL

A

10 – 40 mg/dL (1.1 – 4.4 mmol/L)

127
Q

Lactate value indication for Viral meningitis

A

< 25 mg/dL

128
Q

Lactate value indication for Bacterial meningitis

A

> 35 mg/dL = cut off value 30-36

129
Q

Conversion factor of lactate

A

Conversion factor: mg/dL x 0.1110

130
Q

Elevated levels in existing hepatic encephalopathy

A

Glutamine

131
Q

synthesized from ammonia and glutamic acid serves as the means for CNS ammonia removal.

A

Cerebral glutamine

132
Q

Normal CSF glutamine levels

A

8 – 18 mg/dL

133
Q

MICROBIOLOGICAL EXAMINATION
Basaha lang

A

Normal storage is at room temperature

Gram’s stain sensitivity ranges from 60 – 90% with greatest sensitivity corresponding to higher concentrations of bacteria (10 5 colony forming units/mL).

Patients with PMNs but no organism on gram’s stain, the more sensitive acridine orange stain may be helpful.

Immunochromatographic membrane assay with streptococcus latex agglutination bacterial antigen on H. influenzae, N. meningitis, and S. pneumoniae.

134
Q

T or F
Normal storage is not at room temperature (MICROBIOLOGICAL EXAMINATION)

A

F

135
Q

Stain used for MICROBIOLOGICAL EXAMINATION

A

Gram’s stain

136
Q

Gram’s stain sensitivity ranges from? (%)

A

60 – 90%

137
Q

-Not routinely performed in the laboratory
-It has high sensitivity but low specificity

A

Immunologic studies

138
Q

Rapid detection for meningitis and detects presence of
neurosyphilis but has a very high chance of producing false
positive and false negative results.

A

Immunologic studies

139
Q

(7) But when using immunologic studies in CSF we can use:

A

o Coagulation techniques
o Counterimmunoelectrophoresis
o ELISA
o FTA-Abs
o Latex agglutination
o Radioimmunoassay
o VDRL

aubf lecture final term (5 decks)

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