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FORENSIC MEDICINE COURSE

FORENSIC MEDICINE COURSE. FORENSIC TOXICOLOGY & FORENSIC ALCOHOLOGY. KRZYSZTOF S. BOROWIAK. HISTORICAL ROOTS OF TOXICOLOGY. FORENSIC TOXICOLOGY

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FORENSIC MEDICINE COURSE

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  1. FORENSIC MEDICINE COURSE FORENSIC TOXICOLOGY & FORENSIC ALCOHOLOGY KRZYSZTOF S. BOROWIAK

  2. HISTORICAL ROOTS OF TOXICOLOGY FORENSIC TOXICOLOGY As medicine science was develope in the end of XVIII C. (London, Vien, Berlin). The first Forensic Department in Poland was founded in Krakow in the 1805 CLINICAL TOXICOLOGY First Poisining Centre in Washington D.C. (1957) • EXPERIMENTAL • TOXICOLOGY • Geno • Immuno • new products OCCUPATIONAL AND INDUSTRIAL TOXICOLOGY 60. of XX C.

  3. FORENSIC CLASSIFICATION OF TOXINS POISONS ORGANIC NOT VOLAITABLE SUBSTANCES ISOLATED WITH SOLID 0PHASE OR LIQUID EXTRACTION, detectioned with GC, HPLC, Immuno-methods, spectrometry VOLAITABLE WITH WATER STEAM Detectioned with: GC, GC-MS or Spectrometry NON-ORGANIC SUBSTANCES (METALS) WHEN MINERALISATION IS NECCESSARY, DETECTION WITH AAS

  4. FORENSIC CLASSIFICATION OF TOXINS (2) POISONS EFFECTS CAN BE OBSERVED IN AUTHOPSY ANATHOMOPATOLOGICAL CHANGES ARE NOT OBSERVED Heavy metals Blood toxins: - blocking oxygen transport or metabolism • Drug of abuse • chemicals • Drugs and pharmaceutics • others With characteristic odour Corrosive and irritants

  5. MOST FREQUENT POISONS DETECTED IN THE POST MORTEM SAMPLES 5% 15% 45% 55% ABBREVATION – IN POLISH FORENSIC STATISTIC ON THE 3RD PLACE THE CARBON MONOXIDE IS LOCATED

  6. THE COLLECTION ROUTES AND TYPE OF POST MORTEM SAMPLES When the results of authopsy are unknown and we suspect neccessary of multi-direct toxicological examination • Blood ( 2 x 10 mL) – alcohols & drug abuse, drugs & other organics, • Urine (25-50 mL) – alcohols, drug metabolites, drug of abuse • Brain tissue (50g) – psychotropic substances • Kidney (1) – nophrotoxicity estimation and elimination • Liver (50-100 g) – hepatotoxicity and high conc. of metabolites • Intenstine with contents (50g) • Stomach with separated colected contents • Lungs (50 g) – excelent matherial for solvents and gasses int.

  7. THE COLLECTION ROUTES AND TYPE OF POST MORTEM SAMPLES IMPORTANT TO REMEMBER • BLOOD COLLECTED FOR ETHANOL AND OTHER ALCOHOLS DETERMINATION SHOULD BE TAKEN FROM SURFACE VESSELS (VEINS) • BLOOD COLLECTED FOR CARBOXYHEMOGLOBIN • SHOULD BE TAKEN FROM DEEP LOCATED VESSELS • VIALS WITH SAMPLES COLLECTED FOR VOLAITABLE • SUBSTANCES DETERMINATION SHOULD BE PERFECT • CLOSED • ALL BIOLOGICAL SAMPLES SHOULD BE PRESERVED IN TO ABSOLUTLY CHEMICALY CLEAN VIALS, • BOTLES AND JARS

  8. DIFFERENTIATE OF DEATH CASE Observation during authopsy: pink-redish colour of body surface, internal organ surface and blood • That can be resulted from: • death in the course of carbon monooxide fatal intoxication (COHb > 40% detected in the blood) • death in the course of cianides intoxication • (bitter almond smell) • death in the course of low temperatures action (freezing)

  9. Differentiate course of death Observation durring authopsy: gray-blue colour of corps and organs surfaces, chocolade colour of blood • Result from: • Death in course of methemoglobinemic substances intoxication (nitrates, nitrites and anilin e.g. – MtHb > 30%) • deep anoxemia, metabolic acidosis • putrification of body

  10. OTHER CHARACTERISTIC CHANGES ILLUSTRATED EFFECTS OF POISON INFLUENCE • TYPE OF INJURY EXAMPLES • CAUSTIC POISONS (CORROSIVE – ACUTE LYE BURNS, CORROSION OF GASTRIC MUCOSA, HYPEREMIA, HAEMORAGIAE, NECROSIS) - (STRONG ACIDS, ALKALIAS, HOME SANITARY-CLEANING AGENTS) • SKIN BLISTERS (BARBITURATES) • RENAL DAMAGES & NECROSIS – METALS & METALOIDS • TETRACYCLINES, BABRBITURATES, METHANOL, GLYCOLS, • PULMONARY & GI CHANGES (OEDEMA,HYPEREMIA, HEMORRAGIAE, IRRITANTS,) – GASES, ARSEN OXIDE, INSECTICIDES, PETROLEUM DISTILATE PRODUCTS (GASOLINE, KEROSENE....) • HYPEREMIA – PINK-RED BLOOD AND ORGAN SURFACES – C=O • SKIN AND BLOOD CYANOSIS – METHEMOGLOBINEMIC AGENTS

  11. MECHANISMS OF CELL DEATH FOLLOWING DRUG INDUCED LIVER INJURY (By J.G.O’Grady) DRUG Haptenic epitopes Auto-antigen epitopes Reactive metabolites Covalently modified DNA COVALENT BINDING TO CELLULAR STRUCTURES Mutations, cancerogenesis Covalently modified P-450 Covalently modified proteins DEATH OF CELL Neoantigens Immune system

  12. MORPHOLOGIC CLASSIFICATION OF DRUG-RELATED CHRONIC LIVER INJURY • TYPE OF INJURY EXAMPLES • CHRONIC HEPATITIS HALOTANE, ISONIAZIDE, DANTROLENE, SULPHONAMIDES, • CHRONIC CHOLESTASIS TCA, FENOTHIAZINES, BARBITURATES, PHENYTOIN, TOLBUTAMIDE, • CHRONIC STEATOSIS ETHANOL, METHOTREXATE, ANTINEOPLASTIC AGENTS, • PHOSPHOLIPIDOSIS AMIODARONE, THIORIDAZINE, • VASCULAR (occlusion, ESTROGENS, ANABOLICS, • hypertension, peliosis) • NEOPLASM ESTROGENS, ANABOLICS, VINYL

  13. MORPHOLOGIC CLASSIFICATION OF DRUG-RELATED ACUTE LIVER INJURY (HEPATITIS) • TYPE OF INJURY EXAMPLES • ___________________________________________ • HEPATOCELLULAR(cytolytic) • zonal necrosis - CCL-4, halothane, Acetaminophen • steatosis (makro and mikrovesicular) – ethanol, methotrexate, tetracyclines, valproic acid, • CHOLESTATIC • hepatocanalicular -Amoxiciline, chlorpromazine, • Canalicular – estrogens, steroids, anabolics • VASCULAR(vein occlusion) estrogens, anabolics.

  14. MEDICO-LEGAL ASPECTS OF ETHANOL • Ethyl alcohol is most important constituent of different „alcoholic” beverages (Long-drinks), and basic solvent used in several medicinal preparations. • Ethanol is most frequent abused intoxicating substance • companioned of human from several hundred years. • It is most commonly encountered toxic substance in the • forensic toxicology also. • TYPE OF CRIMES AND ACCIDENTS STRONGLY ASSOCIATED WITH USE OF ETHANOL : • MAJOR TRAUMA • MOTOR VEHICLE COLLISIONS • FIRES, BURNS AND FALLS • DOMESTIC VIOLENCE, CHILD ABUSE • SUICIDES ATTEMPT • TRAUMA AND INJURES OUTCOM COMPLICATIONS

  15. CLINICAL AND MEDICO-LEGAL CLASSIFICATIONOF ETHANOL ACTION > 4,0 - 4,5 ‰ PARALYTIC PHASE (V) 3,0 - 4,0 ‰ NARCOTIC PHASE (IV) 2,0-3,0 ‰ EXCITATION PHASE (III) 1,0-2,0 ‰ EUPHORIC PHASE (II) 0,2-1,0 ‰ DYSPHORIC ACTION (I) < 0,2 - 0,5 > ‰ – STATUS AFTER USAGE OF ALCOHOL - OFFENSE § 87 KW > 0,5 ‰ DRUNKNESS STATUS - CRIME ACCORDING TO § 178 CRIME CODE

  16. STATISTIC DATA OF PAM FOR. DEPARTM. • 2000-2004 YEARS • 240 from 760 clinical examination (alcohols) • 975 drivers examined • 343 under alcohol influence • near 110 under drug abuse (THC, AMPH.) • 84 - both substances • W 2005 - 2007 • Ca. 2800 drivers and victims of accidents were examined (alcohol + drug abuse): 800 -1200 per year • 2007 - under influence of drug abuse 221 examined (from 595) • THC, AMPH, MDMA, Cocaine

  17. AMOUNT OF ETHANOL IN THE DIFFRERENT ALCOHOLIC BEVERAGES • GLASS OF VODKA 100g = 32g PURE SPIRIT • 1-2 GLAS OF 12-14% VINE 500 mL = 32g PURE SPIRIT • 4 GLAS OF PORTER BEER (ALE, GUINESS – 8-9%) = 32g PURE SPIRIT INTAKE ANY OF ABOVE LONG DRINKS CAUSED OVERCROSS THE LEGAL LIMIT OF ETHANOL IN BLOOD OVER 0.5 PROMILLE (0.5 mg/mL)

  18. ETHANOL CONCENTRATION IN SOME COMMERCIAL PRODUCTS

  19. Drinking and driving-the limits in Europe

  20. INFLUENCE OF DIFFERENT FACTORS ON THE BODY-KINETIC OF ETHANOL THEREFORE EFFECTS OF ETHANOL INFLUENCE ARE PERSONAL, INDIVIDUAL AND CHANGEBLE IN TIME

  21. HEPATIC CELLULAR OF ETHANOL BIOTRANSFORMATION

  22. CURVES ILLUSTRATED DIFFERENT BODY-KINETIC OF ETHANOL

  23. EFFECTS OF THE EXCESSIVE ALCOHOLS INTAKE ON THE LIVER TISSUE 1 UNIT = 10 g PURE(spirit) ALCOHOL (100%) 100 ml of vodka (40%)included 32 g of pure alcohol ETHANOL INTAKE,MORE THAN 40 UNITS A WEEK METABOLISM ABNORMALITIES TRIGLICERIDES, SUGARS, GLIKANES ALCOHOLIC HEPATITIS FATTY LIVER PROCESS 70% 30% ALCOHOLIC CIRRORIS FIBROTIC INJURY FATTY LIVER

  24. THE RETROSPECTIVE AND PROSPECTIVE ALCOHOLOMETRIC CALCULATION • WHEN SHOULD BE PERFORMED ? • WHEN DRIVER-PERPETRATOR OF ROAD FATAL ACCIDENT WAS ESCAPED • WHEN ETHANOL ESTIMATION WAS PERFORMED SOME TIME AFTER ACCIDENT • WHEN WHITNESES HAVE DIFFERENT OPPINIONS ON THE SOBRIETY OF DRIVER • WHEN THERE IS POSSIBILITY OF OVERDRINKING OF ETHANOL POST FACTUM • WHEN POSSIBLE ADVERSE OR PATHOLOGIC AFTER ALCOHOL REACTIONS MUST BE RESPECT EXPERT’S OPPINION FOR COURTS, PROSECUTORS AND INSURANCE COMPANIES

  25. RETROSPECTIVE ALCOHOLOMETRIC CALCULATION THEORETIC, CALCULATED CONCENTRATION OF BLOOD ETHANOL, BASED ON THE RESULTS OF ALCOHOLOMETRIC EXAMINATION C(t) = C(a) + B (60) x T When: C(t) – ethanol concentration in blood in the road accident moment (time) C(a) – ethanol concentration estimated in blood in the alcoholometric examination B (60)- elimination valuue factor (0,07-0,28 mg/h) for expert apply limited (0,1-0,2 mg/h) T - time difference from accident to examination (in hours) for example 3 hours

  26. RETROSPECTIVE ALCOHOLOMETRIC CALCULATION continued THE OBLIGATORY CONDITIONS LEAD TO ATTEMPT OF RETROSPECTIVE CALCULATION • THE ETHANOL ELIMINATION PHASE SHOULD BE NOTED (DECREASING RESULT IN THE FOLLOWING EXAMINATIONS e.g. 1.0 ; 0.8 ; 0.7 mg/mL) • TIME PERIOD FROM ACCIDENT TO EXAMINATION NOT OVER 5-7 HOURS • THE ESTIMATED IN BLOOD CONCENTRATION - NOT LESS THAN 0.5 mg/mL

  27. PROSPECTIVE ALCOHOLOMETRIC CALCULATION APPLIED FOR VERYFICATION OF PENETRATOR WITNESS OR OTHER PARTICIPANTS OF ROAD ACCIDENT BASED ON THE DECLARED AMOUNT OF INTAKE ETHANOL • C(o) = A x P x R • C(o) – [mg/ml] theoretical, calculated concentration of blood ethanol based on the known amount and kind of drinken ethanol (declared by driver or whitnes) • A - amount of intake ethanol expressed in the grams of pure (100%) ethanol (e.g. 100 ml 40% vodka = 32 g pure ethanol) • P - body weight (kg) standard range 65-85 kg, at present more frequently BMC (body mas index) is used • R - body distribution coefficient (male - 0.7 ; female – 0.6)

  28. ANALYTICAL METHODS OF ETHANOL DETERMINATION THE PHYSICO-CHEMICAL METHODS DETERMINATION OF ETHANOL IN THE BREATH AIR (used mainly by Police, industry security...) Vessel-pulmonary membrane Ethanol, carbon dioxide Oxygen HENRY LAW (1803) - gases diffusion balance; CUSHING R. (1910) non-active diffussion INFRARED SPECYFIC DETECTORS FOR ETHANOL

  29. ANALYTICAL METHODS OF ETHANOL DETERMINATION cont. Vessel-pulmonary membrane Ethanol, carbon dioxide Oxygen • PERSONAL (ENSURE) – No specyfic, only for drivers autocontrol, may be used also as screening test • PROFFESSIONAL (Police) – different type of detectors • ALCOMAT (Drager), ALCOTEST 7110, ALCOMETR A.2.0 • (estimation of infra-red abssorbance) • ALCOTEST 7310,7410,ALERT, AVAT IV (electrochemical)

  30. ANALYTICAL METHODS OF ETHANOL DETERMINATION cont. Laboratory tube with blood, urine, stomach contents... ETHANOL + Oxidation agent or enzymes Aldehyde and acid acetic, carbon dioxide • GAS CHROMATOGRAPHY (GC) the best procedure: specyfic, fast, multi-detectable, sensitive • GC-MS, GC headspace – mostly use at present • SPECTROMETRIC/ENZYMATIC ( applied ADH for estimation of ethanol) measurement with UV/VIS spectrometers • IMMUNOENZYMATIC (FPIA, EMIT) ABBOT, VIVA

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