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Lab D > Lab 6 Metabolism > Flashcards

Lab 6 Metabolism Flashcards

1
Q

Biuret method

what is it used for and what is the sensitivity

A

To check TP
CuSO4 containing
20-100g/l sensitivity

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2
Q

Average TP concentration of plasma

A

60-80 g/l

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3
Q

How can we measure TP lower than 20 g/l (urine, CSF, etc.)

A 
Lowry method (Folin-phenol)
Ultra sensitive TP method
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4
Q

Biuret test
Chemical reaction
colour of complex
wavelength

A

Photometrical measurement
purple colored complex
546 nm

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5
Q

Ultra sensitive TP analysis
Reagents
wavelength
sensitivity

A

Na-molibdate, pirogallol red
Binding of protein directly to stain
600 nm
0.2-4 g/l

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6
Q

Refractometry mechanism
errors that can give false results
calibration
sensitivity

A

light refraction characteristics of a solution depend on its specific gravity, the TP in blood has the biggest influence on the light refraction

specific gravity depends on temp (20)
hemolysis, lipaemia

distilled water

25-95 g/l

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7
Q

what happens to TP during hyper hydration or dehydration

A

Hyper: decreased TP
De: increased TP

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8
Q 
Protein fraction of dog
Plasma TP
Serum albumin
Serum globulin
Albumin/globulin
Fibrinogen

unit:

A

67-70
48-64% (25-34)
11-21% (30-48)

1.083

1-4

UNIT: g/l

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9
Q

Two methods to measure albumin

A

Spectrophotometry

Electrophoresis

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10
Q

Albumin spectrophotometry

A

Bromocresol green
binds to albumin pH 4.2
Blue-green complex 578 nm

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11
Q

Albumin serum electrophoresis

A

in combination with TP measurement
Protein fraction analysis

Albumin as a % of TP

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12
Q

Decrease of albumin:

intake

A

Decreased digestion or absorption

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13
Q

Decrease of albumin:

synthesis

A

liver failure, acute inflammation

albumin is a negative acute phase protein

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14
Q

Decrease of albumin:

utilization

A

physiological: pregnancy, work, exercise, production (egg,milk)

chronic diseases (infl, neoplasms)

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15
Q

Decrease of albumin:

loss

A 
via kidneys: PLN
via Gi tract: PLE
via skin: burn
whole blood loss
hyperhydration
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16
Q

increase of albumin

A

dehydration

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17
Q

Globulin methods to measure

A

1: calculation: Tp - albumin ≈ globulin

2. serum electrophoresis: % (if TP is known)

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18
Q

Alb/Glob ratio

A

Decrease of ratio: inflammatory/neoplasia (inc globulin)

Inflammation can be analyzed by RBC sedimentation or glutaric-aldehyde test

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19
Q

Electrophoresis basic principles

A

Proteins have amphoteric character (can react both as acid and base)

serum is placed on gel and electric current is exposed to separate proteins into five categories based on size and electric charge

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20
Q

What influences rate of motion during electrophoresis

A

High charge and small size increases rate of motion

Low viscosity also increases speed

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21
Q

What is the efficiency of electrophoretic separation of ions determined by?

A

Relative charge densities (charge per unit volume)

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22
Q

Gel electrophoresis mediums

A

Polyacrylamide or agarose (minimizes diffusion)

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23
Q

SDS-PAGE

A

sodium docecyl sulfate polyacrylamide gel electrophoresis

Proteins denatured by heat
and coated by neg charged SDS
then the protein is applied to the cathode (-) part of the gel and the voltage in the gel forces the neg charged proteins to move toward the anode (+)

densitometer detects results after staining

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24
Q

alpha globulins

A

APP

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25
beta globulins
IgA, IgM | LDL
26
gamma globulins
IgG
27
Monoclonal
One cell group of the same origin
28
Polyclonal gammopathy | ø diseases
increase of globulin conc broad-based peak in beta/gamma region bridging between beta and gamma globulins
29
polyclonal gammopathy diseases bridging
``` Chronic inflammatory disease Liver disease FIP (a2) (virus) Occult heartworm disease (parasite) Ehrlichiosis (bacteria) ``` bridging: increased IgA IgM: lymphoma, heartworm, chronic active hepatitis
30
Monoclonal gammopathy
Sharp spike in beta or gamma region
31
Monoclonal gammopathy neoplasia | 5
``` Multiple myeloma (Plasma cell myeloma) G/A osteolytic bonelesions ``` Lymphoma M/G Chronic lymphocytic leukemia G Extramedullary plasmacytoma in skin of dogs IgM increase: macroglobulinemia B cell lymphoma, spleno-/hepatomegaly
32
monoclonal gammopathy non-neoplastic 6
``` Rare usually IgG (should be ruled out) occult heartworm disease FIPV Ehrlichia vanja Lymphoplasmacytic enteritis/dermatitis Amyloidosis ```
33
Hypoglobulinaemia
Decreased intake: neonates (colostrum, malabs) Decreased synthesis: inherited immunodeficiency, liver function impairment Increased loss: PLN, PLE, skin, bleeding
34
Fibrinogen two methods
1: measurement of both plasma and serum TP (plasma - serum=fbg) 2: plasma TP is measured before and after heat 58 C, refractometry (fbg heat labile) 3: thrombin time
35
Fibrinogen increase
``` Acute inflammation (esp ru) Dehydration ```
36
Fibrinogen decrease
``` liver function impairment, protein deficiency (advanced) DIC Sequestration after body cav bleeding Chronic bleeding blood loss inherited afibrinogenaemia (st bernard) ```
37
Glucometer
Electric conductance (normocytaemia)
38
GOD/POD spectrophotometry
Glu + H2O = gluconic acid + H2O2 = O’ + H2O O’ oxidizes stain from colorless (reduced form) to a colour that depends on glucose conc
39
In vitro catabolism of glucose
``` RbC catabolyzes glu: cooled sample plasma separated quickly coagulate RBC (3cl acetic acid) NaF in blood sample (inhibits enolase) ```
40
Transient glucose increase 6
``` Laboratory errors stress food intake xylazin effect cranial trauma or inflammation glucose containing fluid therapy ```
41
constant hyperglycaemia | 4
Diabetes mellitus Hyperadrenocorticism (cushings), GCS therapy Progesterone Enterotoxaemia of sheep (clostridium)
42
Decreased glu conc 11
``` Lab error (storage/transport) Decreased energy status (ketosis, baby pig, exercise) insulin overdose insulinoma anabolic steroids terminal liver failure acute liver failure hypoadrenocorticism (addisons) septicaemia hyperthyroidism paraneoplastic syndrome ```
43
IV glu tolerance test when is it done and what does it test
during suspicion of DM or insulinoma | 2 starving measurements: >11 mmol/l
44
IV glu tolerance | how is it done
``` animal starved for 24hr blood sampling 0 min 1g/bw 40% glucose conc solution infusion 40 sec) blood sampling at 5, 15, 30, 45, 60 min normalization: 30-60 min ```
45
oral glu tolerance test (glu abs)
suspicion of chronic vowel disease, exocrine pancreatic insufficiency blood sample 0 min 2g/bw 12.5% glucose solution given orally blood sampling 15, 30, 60, 90, 120 twice normal: 30 min normalized: 120 min
46
Macroamylase
glucose bound to alpha amylase | cannot be excreted by kidneys, circulated blood for a longer time
47
fructosamine
glucose bound to more than one protein (ketoamine) conc not influenced by short term hyperglycemia average blood glucose level 2-3 weeks before sampling
48
glycated hemoglobin
non-enzymatic glycation by hgb exposure to plasma glucose | average blood glu levels 2-3 months before measurement
49
Ketone bodies
Acetone, acetoacetate, beta-OH-butyrate decreased intake of carbs or decreased insulin prod
50
Ross reaction
Acetone and acetoacetate conc changes color from white to purple depth depends on ketone conc
51
Relative conc of ketones in different body fluids
Milk: 1 mmol Plasma: 3-5 mmol Urine: 10 mmol
52
Urea of milk and plasma
Can determine energy status of dairy cattle energy deficiency in rumen -> NH3 level increases -> urea production in liver increases milk urea 3 -> 9 plasma urea 9 -> 16
53
What are chylomicrons in the blood a sign off?
post prandial (after eating)
54
How can you differentiate chylomicrons from other lipids in the blood plasma?
Freeze plasma -18, wait 12+hr and slowly warm and centrifuge. the protein part of the CM will coagulate, and leave a transparent layer under the fat. not transparent = lipid mobilization from fat stores
55
causes of hyperlipidaemia 9
``` ponies diet DM hypothyroidism hyperadrenocorticism/gcs therapy nephrotic syndrome septicaemia (energy def) pancreatitis idiopathic (schnauzers, beagles) ```
56
causes of decreased lipid content 3
``` long term starvation liver failure (eg PSS) malabsorption or maldigestion ```
57
Lipid absorption test | when is it done
To determine whether there is existing malabsorption, maldigestion (pancreatic insufficiency) or chronic bowel disease
58
What is normal plasma triglycerol (PT) and what is PT after fast lipid intake in normal conditions?
Normal (dogs) 1 mmol/l | Fast lipid intake 2 mmol/liter
59
how is the lipid absorption test performed?
24hr starvation blood sample 0 min 3ml/bw corn oil po blood sampling each hour if no change: add pancreatic enzyme extract to corn oil if change is seen: epi if still no change: intestinal absorption defect
60
what is cholesterol measurements used for in vet practice?
detection of increased fat mobilization -> total cholesterol value increases (hypothyroidism, hyperadrenocorticism, nephrotic syndrome, DM)
61
what is the fraction of cholesterol-ester of total cholesterol value?
40%
62
what can happen to cholesterol as a result of impaired liver function?
decreased esterificiation | and decreased apolipoprotein production (ldl, hdl, etc)
63
what is the normal value for cholesterol conc?
2-6 mmol/l
64
5 causes of hypocholesterolaemia
``` malnutrition liver failure (decreased synthesis) neoplastic disease hyperthyreosis (inc usage) decreased apolipoprotein synthesis ```
65
7 causes of hypercholesterolaemia
``` high dietary fat content hypothyroidism hyperadrenocorticism DM nephrotic syndrome (plus low TP) cholestatic diseases (bile duct obstruction) idiopathic ```
66
what can cause increased blood FFA conc?
high energy need (lactation)
67
how long can FFA compensate the energy deficiency?
until the liver can not produce any more OAC for beta oxidation
68
starvation, glycogen deficiency of liver, lipid mobilization syndrome and hepatic lipidosis leads to:
decreased TL (total lipid conc) because the liver can not produce enough apolipoproteins for transporting lipids FFA conc is increased because they are transported by albumin
69
normal FFA value
0.1-0.3 mmol/l
70
TL conc
5-7 mmol/l
71
TG conc | sheep?
0.6-1.2 mmol/l sheep has higher 1.5-4 mmol/l
72
cholesterol conc
2-6 mmol/l
73
VLDL
TAG
74
HDL
phospholipids
75
LDL/HDL
Cholesterol
76
(HDL)
cholesterol-ester
77
Albumin transports:
FFA

Lab D (8 decks)

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