BIOMARKER DAN BIOINDIKATOR

1. BIOMARKER DAN BIOINDIKATOR BAHAN KULIAH ANALISIS PENCEMARAN

2. 1.Monitoring • Monitoring must take into consideration the type or types of contaminants present (biological and chemical), their availability and the possibility of biomagnification and bioaccumulation. • The environmental monitoring must be able to detect with accuracy and consistency pollutants present at very low levels.

3. 2.Biomarkers, Test organisms and Bioindicators, • The effects of pollutants on whole organisms representative of the environment, known as bioindicators. • The effects of pollutants on physiological, biochemical, and molecular characteristics of organisms in the environment, known as biomarkers. • The effect of the pollutant on test organisms in the laboratory.

4. Bioindicators and biomarkershave the advantage that they measure the action of the pollutants in the real and complex environment where there may be many and complex interactions at sublethal levels.

5. Definition: Biomarker is a substance used as an indicator of a biologic state Existence of living organisms or biological process. A particular disease state A fragment of DNA sequence BioMarker Proteins Nucleic acids Carbohydrate Lipids Small molecules

6. Location of the biomarker: Location of a particular molecule can also be a marker Biomarkers Cellular-subcellular locations Tissue or organ locations Thymidylate synthase (TS) nucleus vs the cytoplasm The higher the level in NE, lower level in the cyto. Lower survival in colorectal cancer patients Galectin-3 Galectin-3 is a member of lectin family. It is wildly distributed In tissue of epithelial, fibroblast, and dendritic cells. Blood Galectin-3 Predict the outcomes for patients with symptoms of heart failure

7. Detection of biomarker Biomarker Detection of biomarker – diagnosis Self properties, e.g enzymatic activities Antibodies, IHC, ELISA Detection of biomarker Quantitative a link between quantity of the marker and disease Qualitative a link between exist of a marker and disease

8. Ideal Marker for diagnosis Should have great sensitivity, specificity, and accuracy in reflecting total disease burden. A tumor marker should also be prognostic of outcome and predictive of tumor recurrence and effectiveness of anti-cancer treatments. Biomarker & Diagnosis Biomarker for Screening • The marker must be highly specific, minimize false positive and negative • The marker must be able to clearly reflect the early stage of disease • The marker must be easily detected without complicated medical • procedures. The disease markers released to serum and urine are good • targets for application of early screening. • The method for screening should be cost effective. Samples for biomarker detection Blood, urine, or other body fluids samples Tissue samples

12. A.BIOMARKERS • Biomarkers can be defined as quantitative measures of changes in the biological system in response to pollutant exposure. • Biomarkers can be placed into three groups; • biochemical, • immunochemical, and • genetic.

14. Use of Biomarkers in Epidemiology and Clinical Medicine Traditional Exposure Disease Biological or Molecular Epidemiology Markers of ExposureBiomarkers of Disease Exposure dose biological effect Altered clinical prognosis structure/ diagnosis function

15. Predictive biomarkers will allow the selection of lung cancer patients who may need more aggressive screening and treatment Predictive Biomarkers for Lung Cancer Current Status / Perspectives: Although curative resection of patients with early-stage lung CA are performed, the risk of relapse remains substantial Indicates that there may be micro-invasion/metastasis have not been detected by general imaging and/or pathological examinations

16. 2.1.Biochemical biomarkers • Based on the ability of the pollutant to generate a response at the gene level, inducing or increasing specific enzymes involved with detoxification of contaminants. • The detoxification of xenobiotics often involves one of three detoxification strategies: - cytochrome P450, - conjugation with glutathione and -chelation by proteins (Metallothioneines).

17. *EROD, ethoxyresorufin-o-dethylase. ^AHH, aryl hydrocarbon hydroxylase. *GSH, glutathione.polyaromatic hydrocarbon.

18. 2.2.Immunochemical biomarkers • The specific reaction between antigens and antibodies can be used to determine the presence of xenobiotics in environmental samples. • Antibodies against PCBs (polychlorinated biphenyls), PCDDs (polychlorinated dibenzo-p-dioxins), and PCDFs (polychlorinated dibenzofurans) have been developed and used in an ELISA system to determine PCBs in samples (Hahn, 2002).

19. 2.3.Genetic biomarkers Ames test • The Ames test was developed to test substances for their ability to produce mutations in bacteria. The test consists of the treatment of a Salmonella typhimurium histidine auxotroph (His-). The test compound is added to the Salmonella typhimurium His- in an extract of rat liver. • If the compound is a mutagen then mutations will cause revertants- colonies will form on a medium lacking histidine. The number of colonies will give a measure of the mutagenic potential.

21. 2.4.Molecular biology biomarkers • Another development using genetic manipulation of biological material for the estimation of toxicity has been the generation of a transgenic strain of the nematode Caenorhabditis elegans . • The lacZ gene from E. coli is fused to the hsp16 gene. • When the nematode is stressed the enzyme is induced in the worm. The enzyme can be detected by the addition of a substrate such as o-nitrophenyl-p-galactopyranoside (ONPG), which will produce a blue colour when cleaved by the enzyme.

23. 2.5.Fish and Endocrine Disruptors/ Modulators • Fish are good indicators for the aquatic environment • Fish health and fish populations • Commercially and recreationally-important species • Ecologically important species • Indicators of effects on aquatic ecosystem and ultimately human health

24. 2.5.1.Endocrine Modulators • Synthetic or naturally-occurring chemicals that modify or disrupt normal hormonal functions • Alterations in synthesis and metabolism • Improper balance or quantity of circulating hormones • Directly or indirectly interact with natural hormones • Change hormone message and alter cell activity • Induce creation of extra receptor sites • Amplify the hormone effect on cell activity • Block binding sites • Impair normal cell activity • Bind to receptors (hormone mimics) • Abnormal, unpredictable cell activity

25. 2.5.2.Endocrine Disruption • Reproductive • Thyroid • Adrenal (Interrenal, chromaffin) • Neuroendocrine • Immune system/disease resistance • Cortisol • Estrogens/androgens

26. Common Serum Markers for Cancer Diagnosis/prognosis

27. PSA is a protein normally made in the prostate gland in ductal cells that make some of the semen. PSA helps to keep the semen liquid. PSA, also known as kallikrein III, seminin, semenogelase, γ-seminoprotein and P-30 antigen, is a glycoprotein, a serine protease Prostate Cancer marker PSA PSA has several forms in serum Free Pre-enzyme Complex Inactive

28. B.BIOINDICATORS • Bioindicator organisms are those that can be used to identify and quantify the effects of pollutants on the environment. • Lichens are a symbiotic relationship between an alga and fungus and are particularly sensitive to air pollutants and have been used widely used as bioindicators of air quality (Conti and Cecchetti, 2001) • Honey bees have been used as a bioindicator for the contamination of the atmosphere by heavy metals (cadmium, chromium, and lead). • The filter-feeding mussels are known to accumulate heavy metals, making them ideal for the monitoring of coastal waters eg. Mediterranean mussel Myttlus galloprovincialis (Regioli and Principato, 1995). • Frogs, fish eggs as a measure of water quality

29. Bioindikator adalah kelompok atau komunitas organisme yang keberadaannya atau perilakunya di alam berhubungan dengan kondisi lingkungan • Apabila terjadi perubahan kualitas air maka akan berpengaruh terhadap keberadaaan dan perilaku organisme tersebut, sehingga dapat digunakan sebagai penunjuk kualitas lingkungan.

33. Lichens

34. Characteristics • Associated Organism • Thallus (Mass of Hyphae Grasping Algae) • Symbiotic Relationship • Lichenized • Pioneers in New Environments • Long Lifespan (9000 Years Old)

39. Crustose (Crusty) Flat Edges Unlobed Closely Attached to Substrate

42. Foliose (Leafy) Sandwich of Fungal Layer with Algal Mat in Middle Circular Growth Lobes Rootlets Called Rhizines

43. Foliose Lichen

46. Fructicose (Shrubby) Round Branches Vertical Growth Pattern Globets Threads Shrub-like or Mounded Beard-like

50. Leprose (Powdery) Powdery Masses with Little or No Organized Structure