body fluids
TRANSCRIPT
BODY FLUIDS
Mr. Rajesh Kumar Gupta PG Cl. Biochemistry
class ppt
Body Fluids
Extracellular fluid (ECF) Intracellular fluid (ICF)
• plasma, 1/4th • Interstitial fluid, 3/4th
liquid found between the cells or tissue fluidEg. lymph
• Transcellular fluidA body fluid that is not inside cells but is separated from plasma and interstitial fluid by cellular barriers.• CSF• pleural fluid• synovial fluid• Peritoneal fluid, etc
Cerebrospinal fluid (CSF)
Cerebrospinal Fluid (CSF)
• Produced at the Choroid plexus of the 4 ventricles by modified Ependymal cells • At rate @20 ml / hr (adults)
• CSF flows through the Subarachnoid space• Where a volume of 90 – 150 ml is
maintained (adults)• Neonate volume 10-60 mL
• Reabsorbed at the Arachnoid villus / granulation • to be eventually reabsorbed into the
blood
FUNCTIONS OF CSF
▪As shock absorber▪ As mechanical buffer▪ Act as cushion between the brain and cranium▪ Act as a reservoir and regulates the contents of the cranium▪ Serves as a medium for nutritional exchange in CNS▪ Transport hormones and hormone releasing factors▪ Removes the metabolic waste products through absorption
CSF Evaluation
• Tube 1- for cell count and differential• Tube 2- for glucose, protein & enzymes• Tube 3- for culture, Gram stain, AFB stain,
India ink etc• Tube 4- for cytology
Composition of CSF
Appearance Cells DLC Protein glucose
Clear/colorless
0-5/ul(lymphocytes predominant)
∙Adults: 70% lymps, 30% monos.∙Children / newborns: monocyte
15-45 mg/dl 45-75 mg/dl
Typical Viral Meningitis
• CSF WBC elevated, but < 250 (PMNs in early disease, then lymphocytes)
• CSF protein elevated, but < 150 mg/dl• Glucose > 25 mg/dlof serum concentration
Typical Bacterial Meningitis
• CSF WBC > 1000, PMN predominance• CSF protein > 500mg/dl• CSF glucose < 25 mg/dl
Cerebrospinal fluid (CSF)
Biochemical constituents:– Sp. Gravity – 1.003-1.008– CSF pressure – 60-100 mm H2O– pH – 7.28-7.32
Proteins:– 15-45mg/dl– Newborn (0-1mth) – 60-120mg/dl– Albumin/globulin – 3.1
Cerebrospinal fluid (CSF)
Mechanism of increased CSF protein:– increased permeability of the blood brain barrier d/t damage– Decreased reabsorption at the arachnoid villi– Mechanical obstruction of CSF flow due to spinal block above
the puncture site – Increased in inthrathecal immunoglobulin synthesis
• inflammatory meningitis - ↑ to about 125mg-1gm/dl• Neurosyphilis, encephalitis, abscess, tumor - ↑ to 20-
300mg/dl• Spinal cord tumor – 100 – 2000mg/dl
Cerebrospinal fluid (CSF)
Low CSF protein:• May normally occur in young children between 6 months to
12 years• Patient with increased CSF turnover– removal of large volume of CSF– CSF leak induced by trauma or lumbar puncture– increased intracranial pressure, probably due to an
increased rate of protein reabsorption by the arachnoid vili
Cerebrospinal fluid (CSF)
CSF protein estimation:1) Turbidimetric method:– Uses trichloroacetic acid (TCA) or sulfosalicylic acid (SSA) and
sodium sulphate for protein precipitation– Benzethonium chloride or benzalkonium chloride
2) Colorimetric method:– Uses Lowry method (Folin phenol reagent) or– Dye binding method using Coomassie brilliant blue (CBB) or
Ponceau S and– Biuret method
Cerebrospinal fluid (CSF)
Pyrogallol red technique:• Protein present in CSF will quantitatively bind with pyrogallol
red molybdate reagent dye at pH 2.5 violet colored complex
• intensity of this colored complex is measured at 600 nm in a spectrophotometer
• higher the concentration of protein, more intense or darker will be the color of the CSF solution
Cerebrospinal fluid (CSF)
Serum and CSF Albumin &IgG ratio:• assess permeability of blood brain barrier
CSF/Se. albumin index = CSF albumin (mg/dl) Se. albumin (g/dl)• Normal ratio: 1: 230
Cerebrospinal fluid (CSF)
CSF IgG:• 3-5%• MS - ↑ to about 15-18%• Increased intrathecal IgG assessed by
CSF/Se. IgG ratio = CSF IgG (mg/dl) Se. IgG (g/dl)
• Normal ratio: 1:390
Cerebrospinal fluid (CSF)
Cerebrospinal fluid (CSF)Electrophoresis of CSF:• Using cellulose acetate or agarose• Pre-albumin, albumin, alpha1, alpha2, beta1, beta2 and
gammaglobulins• CSF always contain pre-albumin and plasma does not• Pre-albumin or Transthyretin is thyroxine (T4) and retinol-
binding protein
Cerebrospinal fluid (CSF)
• Pattern is abnormal when IgG synthesis increases
• MS – gammaglobulin fraction↑(ologoclonal bands seen)• Multiple sclerosis(MS): is an inflammatory disease in
which the insulating covers of nerve cells in the brain and spinal cord are damaged
• CSF total protein/Gammaglobulin fraction exceeds 0.12 in about 65% of cases of MS
Cerebrospinal fluid (CSF)
High resolution agarose gel electrophoresis:
• Shows discrete patterns of IgG, oligoclonal bands
• Two or more bands necessary for diagnosis
• A highly sensitive stain like silver stain or Coomassie Brilliant Blue is required to identify the proteins in the gel
Cerebrospinal fluid (CSF)
• detection of oligoclonal bands is performed if there is suspicion of an inflammatory or demyelinating condition
• Concomitant serum sample for elecrophoresis and protein estimation is mandatory
• presence of oligoclonal bands in CSF combined with their absence in blood serum often indicates that immunoglobulins are produced in CNS
• Oligoclonal bands are detected in upto 90% of MS
Cerebrospinal fluid (CSF)
• oligoclonal bands are also seen in:
– Panencephilitis - Various viral CNS infection– Neurosyphilis - Neurobrucellosis– Cryptococcal meningitis - Guillian-Barre syndrome– Transverse myelitis - Meningial carcinomatosis– Burkitt’s lymphoma - Chronic relapsing
polyneuropathy– Cysticercosis - Trypanosomasis
Cerebrospinal fluid (CSF)
Glucose:– 50-85mg/dl (2.8-4.4mmol/L), about 2/3rd of plasma value
– normal CSF/plasma glucose ratio = 0.3-0.9
– Hypoglycorrhachia = < 35mg/dl (characteristic of bacterial, tuberculous and fungal meningitis)
– Some viral meningoencephalitis have low CSF glucose but not to that extent as in bacterial meningoencephalitis
Cerebrospinal fluid (CSF)
Decreased level of CSF glucose:– Meningeal involvement in malignant tumor– Sarcoidosis– Cysticercosis– Trichinosis– Amoeba– Acute syphilic meningitis– Intrathecial administration of radoiodinated serum albumin– Subarachonoid haemorrhage– Symptomatic hypoglycemia– Rheumatoid meningitis
Cerebrospinal fluid (CSF)
• Decreased CSF glucose results from increased anaerobic glycolysis in brain tissue and leucocytes and impaired transport to CSF
• CSF glucose normalize before protein levels and cells count during recovery of meningitis, making it a useful parameter in assessing response to treatment
Cerebrospinal fluid (CSF)
Lactate:• 9.0-26mg/dl (1.0-2.9mmol/L)• Elevated CSF lactate reflects CNS anaerobic metabolism due
to tissue hypoxia• Persistently increased – poor prognosis in patient with head
injury• Usually done to differentiate viral from bacterial,
mycoplasma, fungal and tuberculous meningitis where routine parameters yield equivocal results
• Viral meningitis - always <35mg/dl• Bacterial meningitis - >35mg/dl
Cerebrospinal fluid (CSF)
F2 isoprostanes:• Increased in Alzheimer’s disease
Urea:• Level is slightly lower than in blood• In uremia, urea conc. in CSF rises in parallel with that in
blood
Cerebrospinal fluid (CSF)
Enzymes:
Reference range
ADA ( pleural fluid)
Normal: <40 mg/dlSuspect: 40—50 Strong suspect: >50-60Positive: > 60
ADA(CSF) <10 mg/dlADA(Serum)
<15 mg/dl
ADA∙ADA involved in purine metabolism∙It converts Adenosine to inosine∙found mostly in lymphocytes and macrophages
1. Tuberculosis (cut off value > 60 U/L for pleural fluid)
2. ADA is also increased in various infectious disease like
• infectious mononucleosis• Typhoid• Viral hepatitis• Initial stage of HIV• Incase of malignant tumors • SLE
Cerebrospinal fluid (CSF)
Creatine kinase (CK):• Increased CSF CK activity are seen in numerous CSF disorders:– Hydrocephalus, cerebral infraction, primary brain tumors and
subarachnoid hemorrhage
• In patient with head trauma, CSF CK levels correlate directly with the severity of the Concussion
• CK-BB isoenzyme – better than CK-total
Cerebrospinal fluid (CSF)
• CK-BB isoenzyme increases about 6 hour following an ischemic or anoxic insult
CK-BB: < 5U/L - minimum neurological damage 5-20U/L - mild to moderate neurological
damage 21-50U/L - commonly correlated with death
Cerebrospinal fluid (CSF)
LDH:• < 40 U/L• used as the marker in estimating the potential outcome during the
early stages of ischemic brain injury• Also elevated in bacterial meningitis but not in aseptic or viral
meningitis
Ammonia:• Increased levels are generally proportional to the degree of
existing hepatic encephalopathy• Generally correlates with blood value• Also increases in Reye’s syndrome, inherited hyperammonemias
Cerebrospinal fluid (CSF)
Catecholamines:• Homovanillic acid (HVA), the major catabolite of dopamine
and 5-hydroxyindoleacetic acid (5-HIAA), the major catabolite of serotonin, are normally present in CSF
• The levels of both catabolites are reduced in patients with idiopathic or drug induced parkinsonism
Cerebrospinal fluid (CSF)
Tumor marker• Various tumor markers have been seen increased in CSF
of patients with both primary and metastatic tumors• eg. CEA, HCG, ALP
PLEURAL FLUID
Pleural Fluid
• pleural cavity normally contains small amount of fluid that facilitates movement of two membranes against each other
• Plasma filtrate derived from capillaries of the parietal pleura
• Produced continuously at the rate dependent on capillary hydrostatic pressure, plasma oncotic pressure and capillary permeability
• Reabsorbed – lymphatics and venules of visceral pleura
• Volume – about 10ml each
Pleural Fluid
• Accumulation of fluid – an effusion, results from imbalance between the fluid production and reabsorption
• Fluid accumulation in pleural, pericardial and peritoneal cavities serous effusion
Transudate
• Clear, pale yellow, watery substance• Influenced by systemic factors that alter the
formation or absorption of fluid• Increase in hydrostatic pressure • Decrease in plasma oncotic pressure • Contains few protein cells• Common causes: CHF and liver or kidney disease
Exudate• Pale yellow and cloudy substance• Influenced by local factors where fluid absorption is altered
(inflammation, infection, cancer) • Rich in protein (serum protein greater than 0.5)• Ratio of pleural fluid LDH and serum LDH is >0.6• Pleural fluid LDH is more the two-thirds normal upper limit
for serum• Rich in white blood cells and immune cells• Always has a low pH• Common causes: pneumonia, cancer, and trauma
Pleural Fluid
According to Light’s criteria exudate meets one or more of following criteria:
1. Pleural fluid protein/serum protein > 0.52. Pleural fluid LDH/serum LDH > 0.63. Pleural fluid LDH more than two-thirds normal upper
limit for serum
Pleural Fluid
Biochemical constituents:Protein:– contain < 50% of serum protein level– Estimation helps in differentiating transudate or exudate – Protein electrophoresis shows pattern similar to serum
except for higher proportion of albumin
Glucose:– Similar to serum glucose level– Low pleural fluid glucose – malignancy, TB, rheumatoid
pleuritis, non purulent bacterial infections, lupus pleuritis, etc
Pleural Fluid
Lactate:– Useful adjunct - rapid diagnosis of infectious pleuritis
(>90mg/dl)– Levels are significantly high in bacterial and tuberculous
pleural infections
Enzymes:Adenosine Deaminase:– Normal – about 36U/L– Significantly increased in tuberculous pleuritis
Pleural Fluid
Amylase:– Elevations above the serum level (usually 1.5-2 or more
times greater) – pancreatitis, esophageal rupture or malignant effusion
LDH:– Levels rise in proportion to degree of inflammation– Declining LDH level in course of an effusion – resolving of
inflammatory process
Pleural Fluid
Interferon – gamma (INF-gamma):– Useful diagnostic modality for TB pleural effusion– Levels increases significantly in pleural fluid of patient
with tuberculous pleuritis (>136pg/ml)
Lipids:– Pleural fluid Tg level > 110mg/dl – a chylous effusion– Pleural fluid/Se. cholesterol ≥ 0.32 – exudate
Pleural FluidTuberculostearic acid (TSA):– Structural component of Mycobacteriun tuberculosis, not
present normally in human tissue– Using gas chromatography or mass spectroscopy TSA is
measured in sputum, bronchial aspirates, washings or pleural fluid
Tumor markers:– Not recommended routinely– May be useful in diagnosing cases with negative cytology or
unexplained effusions
PERITONEAL FLUID
Peritoneal fluid
• Ascites – pathologic accumulation of excess fluid in the peritoneal cavity
• Normal volume – 50ml• Produced as an ultrafiltrate of plasma dependent on
vascular permeability, hydrostatic and oncotic pressure
Peritoneal fluid
Biochemical constituents:Protein:• serum-ascites albumin gradient or gap (SAAG) SAAG = albumin conc. of serum - albumin conc. of ascitic fluid
• high gradient: – > 1.1 g/dL – due to portal hypertension
• Important causes of high SAAG (> 1.1 g/dL) include:– high protein : heart failure, Budd Chiari syndrome– low protein : cirrhosis of the liver
Peritoneal fluid
• Low gradient:– < 1.1 g/dL - causes of ascites not associated with
increased portal pressure such as tuberculosis, pancreatitis, nephrotic syndrome and various types of peritoneal cancer
Glucose:– Decreased level in tuberculous ascites (< 50mg/dl)– Ascites glucose estimation are of little value
Peritoneal fluid
Enzymes:Amylase:– Amylase activity in normal peritoneal fluid is similar to
plasma level– Level greater than three times the plasma value is
good evidence of pancreas related ascites– Also increases in gastroduodenal perforation, acute
mesenteric vein thrombosis, intestinal strangulation or necrosis
Peritoneal fluid
ALP:– levels >10U/L – predicts hollow visceral injury
LDH:– increased in malignant effusions
Telomerase:– Increased in malignant ascites
Peritoneal fluid
ADA:– Increases in tuberculous peritonitis
Lactate:– Increased in malignant and tuberculous ascites
Creatinine and urea:– Essential in differentiating peritoneal fluid from urine– Increased peritoneal fluid urea and creatinine along with
increased serum urea but normal serum creatinine urinary bladder rupture
Peritoneal fluid
Bilirubin:– Ascitic fluid bilirubin > 6mg/dl and ascitic fluid/serum
bilirubin > 1.0 choleperitoneum
Tuberculostearic acid: helpful Tumor markers: – little value, however, CEA, PSA, α – fetoprotein found to
be very specific for serous fluid malignancies
PERICARDIAL FLUID
Pericardial fluid
• Normal volume: 10-50ml
• Pericardial effusion – excess accumulation
• Often caused by viral infection, most common by enterovirus
Pericardial fluid
• It may be:– transudative (congestive heart failure, myxoedema, nephrotic
syndrome),– exudative (tuberculosis, spread from empyema)– haemorrhagic (trauma, rupture of aneurysms, malignant
effusion).– malignant (due to fluid accumulation caused by metastasis)
• Light’s criteria - reliable diagnostic tool for identifying pericardial exudates and transudates
• Other indicators suggestive of exudate - Specific gravity >1.015, total protein >3.0 mg/dL, LDH >300 U/dL, glucose fluid-to-serum ratio < 1
Pericardial fluid
Biochemical constituents:• Biochemical parameters for the diagnosis of pericardial
effusions have not been studied to the same extent as in other body fluids
Glucose:– Value < 40mg/dl (2.22mmol/L) – bacterial, tuberculous,
rheumatic or malignant effusion
Pericardial fluid
Enzymes:LDH:– Level > 200U/L suggests pericardial exudate
• Significantly increased pericardial fluid levels of CK-MB, myoglobin and Troponin I in postmortem pericardial fluid – myocardial injury
ADA :– Useful adjunctive test for tuberculous pericardits
Pericardial fluid
Interferon-gamma:– Increased in tuberculous pericarditis– Cutoff value – 200pg/L
PCR:– More specific than ADA in diagnosing tuberculous
pericarditis but– Negative test does not rule out tuberculous pericarditis
since some pericardial fluids from patients with large tuberculous effusions may not contain M. tuberculosis
SYNOVIAL FLUID
SYNOVIAL FLUID
• Ultrafiltrate of blood plasma combined with hyaluronic acid produced in the joints space by the synovial cells lining synovial tendon sheaths, joints, etc
• Composition similar to plasma as small ions and molecules readily pass into the joint space
• Reabsorption – lymphatics
• Acts as a lubricant and adhesive, and provides nutrients for the avascular articular cartilage
SYNOVIAL FLUID
• Examination of synovial fluid is essential to differentiate infectious from non-infectious arthritis
Biochemical parameters:• Adds only supportive information to the routine test
Mucin clot test:• Add acetic acid to SF precipitates hyaluronate into a mucin clot
which may be graded as good, fair or poor• Fair to poor mucin clot reflects dilution and depolymerization of
hyaluronic acid, a non-specific finding of several inflammatory arthrites
SYNOVIAL FLUID
Glucose:• Proper interpretation of SF glucose values requires comparison
with serum levels, ideally preceded by eight hours fast to allow glucose to equilibrate across the synovial membrane
• Normally, Serum – synovial = < 10mg/dl, also in many non-inflammatory conditions
• In septic arthritis, this difference increases from 20-60mg/dl
SYNOVIAL FLUID
Protein:• Mean normal – 1.0-3.0 g/dl• Total protein estimation is not generally useful• With increasing inflammation, larger proteins enter the synovial
space
Enzymes:LDH:• Increased in RA, gout, failed arthroplasties and infectious arthritis
reflecting neutrophilic infiltration
SYNOVIAL FLUID
Acid phosphatase:• Elevated acid phosphatase may have negative prognostic
value in RA but is non-specific
Organic acids:Lactic acid:• increased in septic arthritis• Increased > 30mg/dl septic arthritis due to gram +ve cocci
and gram –ve bacilli
SYNOVIAL FLUID
• Using gas-liquid chromatography, presence of other organic acids not normally present in SF (eg. n-valeric, n-hexanoic and succinic acids) may be helpful in differentiating septic from non-septic arthritis
Uric acid:• Increased SF uric acid level supports a diagnosis of gout
Lipids:• Contains extremely low conc. of lipids than in plasma• Helps when cholesterol crystals of SF resemble MSU or CPPD
Reference intervals for synovial fluid constituents
Constituents Synovial fluid Plasma
Total protein 1 - 3 g/dl 6 – 8 g/dl
• albumin 55 – 70 % 50 – 65 %
• α1- globulin 6 - 8 % 3 – 5 %
• α2- globulin 5 – 7 % 7 – 13 %
• β – globulin 8 - 10 % 8 – 14 %
• γ – globulin 10 - 14 % 12 – 22 %
Hyaluronic acid 0.3 - 0.4 g/dl -
Glucose 70 - 110 mg/dl 70 – 110 mg/dl
Uric acid 2 – 8 mg/dl 2 – 8 mg/dl
Lactate 9 - 29 mg/dl 9 – 29 mg/dl
AMNIOTIC FLUID
AMNIOTIC FLUID
• fluid surrounds, protects, and nourishes a growing fetus during pregnancy
• allows the baby to move relatively freely and helps maintain a stable temperature
• increases in volume as fetus grows
• Highest – 34wks
• At first, it is mainly water with electrolytes, by about 12-14th week - proteins, carbohydrates, lipids and phospholipids and urea(all of which aid in the growth of the fetus)
AMNIOTIC FLUID
• detect and diagnose some birth defects, genetic diseases, and chromosome abnormalities in a fetus, especially if pregnancy screening tests are abnormal
• to evaluate fetal lung maturity
• obtained through a procedure - amniocentesis
AMNIOTIC FLUID
• 15 and 20 weeks - for genetic diseases, chromosome abnormalities and open neural tube defects
• after 32 weeks - to evaluate fetal lung maturity, when there is an increased risk of premature delivery
AMNIOTIC FLUID
• For genetic testing and chromosome analysis, fetal cells in the amniotic fluid are cultured and grown for several days in the laboratory, then are analyzed
• Biochemical tests, such as bilirubin and alpha-fetoprotein, and sometimes genetic tests can be performed directly on the amniotic fluid
AMNIOTIC FLUID
• Chromosome analysis, a cytogenetics test that may also be called karyotyping - detect chromosome abnormalities associated with a variety of disorders. (It evaluates the 22 paired chromosomes and the sex chromosomes (XY) in the nucleus of cells cultured from those collected in the sample of amniotic fluid and can be used to diagnose a variety of chromosomal disorders)(down’s, klinefelter, edward, patau’s, turner)
• Genetic testing, also called molecular testing.( It looks at fetal DNA to identify specific gene mutations and diagnose a variety of inherited diseases)(cystic fibrosis, tay sach’s disease, sickle cell anemia, thalessemia)
AMNIOTIC FLUID
• AFP (alpha-fetoprotein)—increased with neural tube defects
• Acetylcholinesterase—increased with neural tube defects and also other anatomic abnormalities
• Testing to evaluate fetal lung maturity- (tests are based upon the presence of adequate protective liquid substances called surfactants in the lungs, which are necessary for proper lung function)
• Tests for bilirubin may be performed on a regular basis, starting at about 25 weeks of pregnancy, to detect, evaluate and monitor the severity of the hemolytic anemia in the fetus
SEMINAL FLUID
SEMINAL FLUID
• Semi-gelatinous or liquid suspension containing spermatozoa and secretions from male accessory organ
• Net fluid formed by mixing of the testicular fluid, prostatic fluid and secretion from the seminal vesicles
• Composition is most suitable for the maintenance and survival of
spermatozoa
SEMINAL FLUID
• Has same pH as blood plasma
• Conc. of lactate, phosphate and citrate is higher than in blood
• Chloride and cholesterol are lower
• Sugar content is high –fructose
• If fructose is low – infertility (spermatozoa can’t survive)
SEMINAL FLUID
• semen analysis is used to determine whether a man might be infertile
Components Reference rangeVolume (ml) 2.3 – 2.99pH 7.19 – 8.47Osmolality (mosm) 254 – 423Fructose (mg/dl) 136 – 628Glucose (mg/dl) 5 – 295Total protein (mg/dl) 3700 – 7460Albumin (mg/dl) 1100 – 2000Urea (mg/dl) 13 – 98Lactic acid (mg/dl) 22 – 136Citrate (mg/dl) 304 – 751Ca (mg/dl) 16 – 53Cl (mg/dl) 130 – 158K (mg/dl) 50 – 248Mg (mg/dl) 7.89 – 31.8Na (mg/dl) 236 – 512Zn (mg/dl) 6.78 – 69.29
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