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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
Q

beta globulins

A

IgA, IgM

LDL

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26
Q

gamma globulins

A

IgG

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27
Q

Monoclonal

A

One cell group of the same origin

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28
Q

Polyclonal gammopathy

ø diseases

A

increase of globulin conc
broad-based peak in beta/gamma region
bridging between beta and gamma globulins

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29
Q

polyclonal gammopathy
diseases
bridging

A 
Chronic inflammatory disease
Liver disease
FIP (a2) (virus)
Occult heartworm disease (parasite)
Ehrlichiosis (bacteria)

bridging:
increased IgA IgM:
lymphoma, heartworm, chronic active hepatitis

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30
Q

Monoclonal gammopathy

A

Sharp spike in beta or gamma region

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31
Q

Monoclonal gammopathy neoplasia

5

A 
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
Q

monoclonal gammopathy
non-neoplastic
6

A 
Rare
usually IgG (should be ruled out)
occult heartworm disease
FIPV
Ehrlichia vanja
Lymphoplasmacytic enteritis/dermatitis
Amyloidosis
33
Q

Hypoglobulinaemia

A

Decreased intake: neonates (colostrum, malabs)

Decreased synthesis: inherited immunodeficiency, liver function impairment

Increased loss: PLN, PLE, skin, bleeding

34
Q

Fibrinogen two methods

A

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
Q

Fibrinogen increase

A 
Acute inflammation (esp ru)
Dehydration
36
Q

Fibrinogen decrease

A 
liver function impairment, protein deficiency (advanced)
DIC
Sequestration after body cav bleeding 
Chronic bleeding
blood loss
inherited afibrinogenaemia (st bernard)
37
Q

Glucometer

A

Electric conductance (normocytaemia)

38
Q

GOD/POD spectrophotometry

A

Glu + H2O = gluconic acid + H2O2 = O’ + H2O

O’ oxidizes stain from colorless (reduced form) to a colour that depends on glucose conc

39
Q

In vitro catabolism of glucose

A 
RbC catabolyzes glu:
cooled sample
plasma separated quickly 
coagulate RBC (3cl acetic acid)
NaF in blood sample (inhibits enolase)
40
Q

Transient glucose increase 6

A 
Laboratory errors
stress
food intake
xylazin effect
cranial trauma or inflammation 
glucose containing fluid therapy
41
Q

constant hyperglycaemia

4

A

Diabetes mellitus
Hyperadrenocorticism (cushings), GCS therapy
Progesterone
Enterotoxaemia of sheep (clostridium)

42
Q

Decreased glu conc 11

A 
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
Q

IV glu tolerance test when is it done and what does it test

A

during suspicion of DM or insulinoma

2 starving measurements: >11 mmol/l

44
Q

IV glu tolerance

how is it done

A 
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
Q

oral glu tolerance test (glu abs)

A

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
Q

Macroamylase

A

glucose bound to alpha amylase

cannot be excreted by kidneys, circulated blood for a longer time

47
Q

fructosamine

A

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
Q

glycated hemoglobin

A

non-enzymatic glycation by hgb exposure to plasma glucose

average blood glu levels 2-3 months before measurement

49
Q

Ketone bodies

A

Acetone, acetoacetate, beta-OH-butyrate

decreased intake of carbs or decreased insulin prod

50
Q

Ross reaction

A

Acetone and acetoacetate conc
changes color from white to purple
depth depends on ketone conc

51
Q

Relative conc of ketones in different body fluids

A

Milk: 1 mmol
Plasma: 3-5 mmol
Urine: 10 mmol

52
Q

Urea of milk and plasma

A

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
Q

What are chylomicrons in the blood a sign off?

A

post prandial (after eating)

54
Q

How can you differentiate chylomicrons from other lipids in the blood plasma?

A

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
Q

causes of hyperlipidaemia 9

A 
ponies
diet
DM
hypothyroidism
hyperadrenocorticism/gcs therapy
nephrotic syndrome 
septicaemia (energy def)
pancreatitis 
idiopathic (schnauzers, beagles)
56
Q

causes of decreased lipid content 3

A 
long term starvation
liver failure (eg PSS)
malabsorption or maldigestion
57
Q

Lipid absorption test

when is it done

A

To determine whether there is existing malabsorption, maldigestion (pancreatic insufficiency) or chronic bowel disease

58
Q

What is normal plasma triglycerol (PT) and what is PT after fast lipid intake in normal conditions?

A

Normal (dogs) 1 mmol/l

Fast lipid intake 2 mmol/liter

59
Q

how is the lipid absorption test performed?

A

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
Q

what is cholesterol measurements used for in vet practice?

A

detection of increased fat mobilization -> total cholesterol value increases
(hypothyroidism, hyperadrenocorticism, nephrotic syndrome, DM)

61
Q

what is the fraction of cholesterol-ester of total cholesterol value?

A

40%

62
Q

what can happen to cholesterol as a result of impaired liver function?

A

decreased esterificiation

and decreased apolipoprotein production (ldl, hdl, etc)

63
Q

what is the normal value for cholesterol conc?

A

2-6 mmol/l

64
Q

5 causes of hypocholesterolaemia

A 
malnutrition
liver failure (decreased synthesis)
neoplastic disease
hyperthyreosis (inc usage)
decreased apolipoprotein synthesis
65
Q

7 causes of hypercholesterolaemia

A 
high dietary fat content 
hypothyroidism
hyperadrenocorticism 
DM
nephrotic syndrome (plus low TP)
cholestatic diseases (bile duct obstruction)
idiopathic
66
Q

what can cause increased blood FFA conc?

A

high energy need (lactation)

67
Q

how long can FFA compensate the energy deficiency?

A

until the liver can not produce any more OAC for beta oxidation

68
Q

starvation, glycogen deficiency of liver, lipid mobilization syndrome and hepatic lipidosis leads to:

A

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
Q

normal FFA value

A

0.1-0.3 mmol/l

70
Q

TL conc

A

5-7 mmol/l

71
Q

TG conc

sheep?

A

0.6-1.2 mmol/l
sheep has higher
1.5-4 mmol/l

72
Q

cholesterol conc

A

2-6 mmol/l

73
Q

VLDL

A

TAG

74
Q

HDL

A

phospholipids

75
Q

LDL/HDL

A

Cholesterol

76
Q

(HDL)

A

cholesterol-ester

77
Q

Albumin transports:

A

FFA

Lab D (8 decks)

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