SEMEIOTICA di LABORATORIO

ALGORITMO DIAGNOSTICO IPOCALCEMIA

ALGORITMO DIAGNOSTICO IPERCALCEMIA
 BONE ALKALINE  Enzyme present in osteoblast
PHOSPHATASE plasma membranes
 Enzymatic degradation of the
mineralisation inhibitor
pyrophosphate at alkaline pH
 ADVANTAGES. Low intra-
individual variability <10% Not
affected by renal function, Food
has little effect Long circulatory
half-life 1–2 days ; Sample
stability;Cheap
 DISADVANTAGES. Up to 20%
cross reactivity with liver
Isoforms; Changes with therapy
minimal ; 2 peaks at 14:00 and
23:30 hrs Nadir 30% ↓at 06:30;
Multiple methodologies, can
measure mass or activity.

BONE ALKALINE PHOSPHATASE

 OSTEOCALCIN  Major non-collagen bone protein.
Produced by osteoblasts during bone
formation and bound to hydroxyapatite

 Influences osteoid mineralisation

provides negative feedback during
remodelling process. Late marker of
osteoblast activity

 Intact molecule unstable. Large inter-lab

variation . Short half-life of a few
minutes . Influenced by Vit K status,
renal function and circadian variability.

 OC gene regulated at transcriptional

level by 1,25-OH2 Vit D. Vit K essential
cofactor for γ-carboxylation of OC
resulting in ↑ affinity for Ca and
hydroxyapatite.

OSTEOCALCIN
 HYDROXYPROLINE is produced by
hydroxylation of the amino acid proline
by the enzyme prolyl hydroxylase
following protein synthesis (as a post-
translational modification
 Hydroxyproline is a major component of
the protein collagen.Hydroxyproline and
proline play key roles for collagen
stability. They permit the sharp twisting
of the collagen helix
 Proline hydroxylation requires ascorbic
acid. The first effect of absence of
ascorbic acid in humans comes from the
resulting defect in hydroxylation of
proline residues of collagen, with
reduced stability of the collagen
molecule, causing scurvy.
 Increased serum and urine levels of
hydroxyproline have also been
demonstrated in Paget's disease and
during bone remodelling/resorption (high
values of hydroxyproline/creatinine ratio)

HYDROXYPROLINE
 CTX (telomero amino-terminale),
NTX (telomero carbossi-terminale)
 Modalità di prelievo e Tecnica
analitica Analisi immunoenzimatica o
ELISA sulle urine delle 24 ore
(Serum CrossLaps) per la
determinazione del NTX. Analisi in
HPLC o RIA per la determinazione
del CTX.
 Unità di misura nmol BCE (Bone
Collagen Equivalent) mmol/mmol
creatinina
 Valori riferimento
 NTX: Maschi 0-85 mmol/mmol
creatinemia; Femmine 14-76
mmol/mmol creatinemia
 CTX: Adolescenti: 150 e 400
nmol/mmol creatinina. Adulti: Maschi
20-40 nmol/mmol creatinina;
Femmine 30-90 nmol/mmol
creatinina.
TELOPEPTIDI AMINO- E CARBOSSI-TERMINALE DEL COLLAGENE
 I Telopeptidi amino- e carbossi-terminale del collagene
markers di turnover del metabolismo osseo e
rappresenta un indice tra i più affidabili del
riassorbimento osseo. I cross-link che coinvolgono
telopeptidi e gli acidi coniugati della piridina si formano
durante la maturazione del collagene di tipo I che
costituisce circa il 90% della matrice dell’osso. Il
collagene-I è una tripla elica di due catene
amminoacidiche identiche. Nel corso della formazione
dell’elica si vengono a costituire cross-link N- e C-
telopeptidici tra i residui idrossilisinici 1 e 2 presenti
sulle neurofibrille che incorporano desossipiridinolina
(DPD) e piridinio (PYD). La sequenza nota come
telopeptide amino-terminale (NTX) vie a liberarsi a
seguito dell’attività idrolitica della Catepsina K
prodotta dagli osteoclasti. Il telopeptide carbossi-
terminale (CTX) – ricco in residui desossipiridinolinici -
Nel corso del riassorbimento osseo frammenti delle
catene contenenti i telopeptidi e i residui
desossipiridinolinici vengono immessi in circolazione e
quindi filtrati ed escreti nelle urine