salivary markers of systemic diseases ppt
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SALIVARY MARKERS OF SYSTEMIC DISEASES
BY
Y.PRAGZNA(10T21S0113)
UNDER THE GUIDANCE OFMrs. M. SANDHYA M.PHARM
ASSOCIATE PROFESSOR
CONTENTS:
Biomarkers
Saliva physiology
Advantages & disadvantages
Saliva collection, processing & storage
Diagnostic applications
Salivary proteome & transcriptome
New technologies of measurement
BIOMARKERS:
Biomarkers are defined as cellular, biochemical and molecular characteristics by which normal/abnormal processes can be recognized and/or monitored.
SALIVARY MARKERS
Non invasive diagnosis of disease and monitoring of general health
SALIVA PHYSIOLOGY:
Saliva is the mixed product of 3 major salivary glands.
Parotid
Sub mandibular
Sub lingual
It is slightly acidic(pH 6-7).
Several physiological & pathological conditions modify saliva production quantitatively.
FUNCTIONS OF SALIVA
PROTECTIVE Lubrication Anti-microbial Mucosal integrity Lavage/cleansing Buffering Remineralization
FOOD & SPEECH RELATED
Food preparation Digestion Taste Speech Prevention of
caries
SA
LIV
A A
S A
D
IAG
NO
STIC
FLU
ID
AD
VA
NTA
GES
DIS
AD
VA
NTA
GES
It is safe, non-invasive and simple.
No needles and No cringing.
Home testing.
It has an average protein concentration of 1.5-2 mg/ml.
Cost of shipping & Storage tend to be lower.
Informative analytes are present in lower amounts in saliva than in serum.
SA
LIV
A C
OLLEC
TIO
N
WHOLE SALIVA COLLECTION
DUCTAL SECRETIONS COLLECTION
PAROTID SECRETIONS COLLECTION
SUB MANDIBULAR & SUB LINGUAL SECRETIONS COLLECTION
PROCESSING & STORAGE:
All procedures must be performed on ice:
(1)Divide the saliva samples into multiple 330μL samples placed in cryotubes at -80ºc.
Further processed for Protein analysis: Add protease inhibitors like aprotinin(0.33µL), 3.3µL PMSF.
RNA analysis: Add 1.65µL of SUPERase inhibitor.
(2)Invert gently to mix.
(3)Store all fractions at -80ºc.
Salivary/Oral Fluid Biomarkers Possibilities of Use
DNA Standard genotyping Genetic information of the hosting human
body Bacterial infection (oral microbes in the mouth)
Diagnosing carcinomas of head & neck Forensics
mRNA Information on transcription rates
RNA Viral/bacterial identification
Proteins Genetic information Translational regulation Diagnosing periodontitis Detecting dental cavities
Immunoglobulin Diagnosing viruses
Metabolites Diagnosing periodontitis
DIAGNOSTIC APPLICATIONS OF SALIVA:Drugs, hormones:
☻Intra cellular pathways Active & Passive diffusion
☻Extra cellular pathways Ultra filtration
Serum components:
☻Capillary barrier☻Interstitial spaces☻Cervicular fluid☻Membrane of acinar & ductal cells
SJOGRENS SYNDROME: There are raised concentrations of NaCl, IgA, IgG,
lactoferrin, inflammatory mediators such as PGE2, TXB2,
Interleukin-6 etc.
CYSTIC FIBROSIS: Saliva of CF patients contain increased calcium &
phosphate levels, urea & uric acid and abnormally elevated
levels of PGE2.
DIABETES MELLITUS: It effects flow rate & composition of saliva. Salivary
alterations includes higher levels of IgA, Salivary Peroxidase, glucose content, K, Salivary total protein &
amylase.
ORAL MUCOSAL DISEASES: Nitric oxide levels were increased.
ONCOLOGY:
P53 antibody can be detected in oral squamous cell carcinoma, increased levels of salivary kallikrein levels in oral malignant tumors, CA 15-3 in breast cancer.
CVS:
Determination of salivary amylase activity before 6hrs after cv surgery.
KIDNEY DYSFUNCTION:
Hypo salivation , changes in taste, ammonium smelling breath, oral mucosal pain.
EPILEPSY:
Gingival hypertrophy, IgA deficiency
VIRAL DISEASES:
Acute hepatitis A & B were diagnosed based on the presence of IgM antibodies in saliva.
HIV:
Salivary IgA levels.
BACTERIAL INFECTIONS:
By monitoring the presence of antibodies to microorganisms found in saliva & oral cavity
AGAIN & AGAIN ABOUT TOBACCO:
Smoking causes decrease in salivary important antioxidant levels.Salivary nicotine levels are considered to be the most reliable marker
DR
UG
MO
NIT
OR
ING
ANTI-ASTHMATICS: Theophylline
ANTI EPILEPTICS: Carbamazapine, Diazepam, Ethosuximide, Lamotrigine, Topiramate, Phenytoin, Primidone, Valproic acid
ANTI ARRYTHMIC/ANTI HYPERTENSIVES: Digoxin, Metoprolol, and Procainamide.
ANTI MICROBIALS: Clarithromycin, Gentamycin, Isoniazid, Ofloxacin, Sulfanilamide
ANTIPYRETICS: Acetaminophen, Antipyrine, Paracetamol
ANTI NEOPLASTICS: Cisplatin, Methotrexate, Texol, Topotecan
ANTI VIRAL: Quinine
LITHIUM PSYCHOTROPIC: Lithium, Amitriptyline
MISCELLANEOUS: Nicotine, Ethanol, Caffeine, Tolbutamide, Quinine
ILLICIT DRUGS: Amphetamine, Barbiturates, Benzodiazepines, Opiods, Cocaine, Phencyclidine
HORMONE MONITORING:Steroids like cortisol, aldosterone, testosterone, insulin
SIALOCHEMISTRY ANALYSIS:Early monitoring of an exposure to lead, cadmium, mercury because of higher salivary levels of these elements
CLASSIFICATION OF SALIVARY BIO MARKERS:
◘PROTEOMIC
◘GENOMIC
◘MICROBIAL
◘OTHERS
DIA
GN
OS
TIC
MO
LEC
ULA
R
TAR
GETS
SALIVARY PROTEOME:
Use of ‘Divide & conquer’ strategy
Initial fractionation of proteins with separation techniques
Digestion with proteolytic enzyme, trypsin
Analysis by 1-D, 2-D LC-MS
Finally data is processed & submitted for database searching
SA
LIVA
RY P
RO
TEO
MIC
AN
ALY
SIS
SALIVARY TRANSCRIPTOME:
◘It includes mRNA molecules that cells use to convey the instructions carried by DNA for subsequent protein production.
◘At present, the main strategy for identification of salivary transcriptomic biomarkers is through microarray technology.
SA
LIVA
RY T
RA
NS
CR
IPTO
ME:
DNA MICROARRAY TECHNOLOGY
NEW TECHNOLOGIES FOR MEASURING SALIVARY BIOMARKERS:
MICROBEAD ARRAY
Micro pits within a
silicon wafer are populated with a variety of chemically sensitized bead micro reactors.
POC DEVICE
In this POC device, saliva(100-300μL) is placed into the salivary collection module, and then delivered into the Nano-biochip.
MIC
RO
BEA
D A
RR
AY T
EC
HN
IQU
E NANO-BIOCHIP
CONCLUSION:
The components of saliva act as a “Mirror of the body’s health”, widespread use and growing acceptability of saliva as a diagnostic tool is helping individuals, researchers, health care professionals and community health programs to better detect and monitor diseases and to improve the general health of the public
REFERENCES:◘Mandel ID. A contemporary view of salivary research. Crit Rev Oral Biol Med 1993; 4:599.
◘Raymond G, Schipper a, Erika Silletti a, Monique H, Vingerhoeds. Saliva as research material: biochemical, physiochemical, and practical aspects. Archives of Oral Biology 2007 ;( 52):1114-35.
◘Richard P, Jiri S, Jana V, Edgar F, Petr M, Jindrich P et al. Saliva as a diagnostic medium. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2009; 153(2):103-10.
◘Eliaz Kaufman .The diagnostic applications of saliva. A review. Crit Rev Oral Biol Med 2002; 13(2):197-212.
◘www.med.kufauniv.com/ar/journals2010/res%20(1).pdf
◘www.jofs.in/files/Artical-11.pdf
◘www.linkgroup.hu/docs/pcs2/218-08CurrGen.pdf
◘www.wikipedia.com
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