Marija Definis-Gojanović

Identification of War Victims from Mass Graves in Croatia, Bosnia, and Herzegovina by the Use of Standard Forensic Methods and DNA Typing

The postmortem remains of sixty-one war victims were excavated from 6 mass graves in Bosnia and Herzegovina one and a half years after interment Using standard identification methods, including the matching of medical and dental records, the recognition of distinguishing characteristics such as the use of clothing and belongings, and video superimposition, 35 persons were identified. For the remaining 26 persons identification efforts continue. DNA typing was performed at the HLA DQA1 locus and five PM system loci. Results from DNA typing were confirmed by other methods. DNA profiles of family members of 150 missing persons are now being developed using the 6 loci. These DNA profiles will then be compared with those generated from the bone and teeth remains of the unidentified victims.

Interaction of humic acids with human DNA: Proposed mechanisms and kinetics

Human DNA quantification by quantitative real‐time PCR (QRT‐PCR) has gained great importance in forensic DNA and ancient DNA studies. However, in such samples, DNA quantification is impaired by the frequently present humic acid (HA). We have previously shown that the addition of synthetic HA inhibits QRT‐PCR. In this study we investigated the possible mechanisms of HA interaction with human DNA, and kinetics of QRT‐PCR inhibition. In QRT‐PCR with pure human DNA and no HA added, VMAX was 40. With DNA sample containing 4 μg/mL of HA, VMAX was 30.30 while the addition of extra Taq polymerase to the same sample changed VMAX into 38.91, amplifying between 80 and 90% of input DNA. The KM/VMAX ratio in all the samples remained constant, indicating that the mechanism of HA inhibition of QRT‐PCR is uncompetitive by nature. Moreover, HA shifts the human DNA melting temperature point (Tm) from 75 to 87°C and inhibits DNase I‐mediated DNA cleavage, most probably affecting the enzymes activity.

Northern and southern Croatian population data on seven PCR-based loci

Northern and southern Croatian sample populations were typed at seven PCR-based loci -LDLR, GYPA, HBGG, D7S8, Gc, HLA-DQA1 and D1S80. The results show that all loci meet Hardy-Weinberg expectations and that there is little evidence for association of alleles between loci. Allelic frequency distributions at all loci, except HLA-DQA1, show no differences between the northern and southern Croatian sample populations. Moreover, the population data for Croatians are similar to U.S. Caucasians; only HLA-DQA1 for southern Croatians was statistically different compared with U.S. Caucasians. A Croatian population database(s) has been created and can be used for forensic analyses to estimate the frequency of a multiple locus DNA profile.

Suicide by fentanyl.

Suicide by Fentanyl Fentanyl is a potent, short-acting narcotic analgesic widely used as surgical anaesthetic. This article presents a case in which fentanyl was self-injected by a 41-year old nurse, an employee at the hospital emergency department, who was found dead at home. She had no known history of drug and alcohol abuse. Two syringes, one empty and one filled with a clear liquid, were found near the body, while a needle was stuck into her hand. Toxicological analysis showed fentanyl poisoning. Fentanyl overdose was declared the cause of death and the manner of death was classified as suicide. To our knowledge, death due to the intravenous injection of fentanyl has not previously been reported in Croatia. Samoubojstvo Fentanilom Fentanil je snažan narkotik u velikoj mjeri upotrebljavan kao anestetik pri kirurškim zahvatima. U ovom radu prikazan je slučaj samoubojstva upotrebom fentanila. 41-godišnja medicinska sestra, zaposlena u hitnoj službi pronađena je mrtva u svom stanu. Nije bilo podataka o njezinu zaživotnom uživanju alkohola niti drugih sredstava ovisnosti. Dvije šprice, jedna prazna i druga ispunjena prozirnom tekućinom, pronađene su kraj njezina tijela, dok je igla bila ubodena u njezinu ruku. Toksikološka analiza dala je pozitivan rezultat na fentanil. Uzrok smrti bio je predoziranje fentanilom u svrhu samoubojstva. Prema našim saznanjima, ovo je prvi dokazani slučaj samoubojstva u Hrvatskoj koji je počinjen fentanilom.

Drug analysis in necrophagous flies and human tissues.

Drug Analysis in Necrophagous Flies and Human Tissues Necrophagous insects may provide useful information about the time, place and cause of death. In addition, they can serve as reliable alternative specimens for toxicological analysis in cases where human tissue and fluids, normally taken during autopsies, are not available, due to decomposition of the corpse. This paper reports the results of drug analysis of the larvae of two fly families, Calliphoridae and Sarcophagidae, collected from the body of a middle-aged man who had committed suicide approximately three weeks before his corpse was found. Multiple samples of decomposed human tissue, of the blowfly, and of the larval flesh were analysed using gas chromatography/mass spectrometry (GC/MS), and amphetamine was detected in all samples. While the screening results were beyond doubt, the quantitative analysis was less clear, and further research is needed in this area. Analiza Droga U Nekrofilnim Muhama I Ljudskim Tkivima Insekti strvinari mogu pružiti korisne informacije o vremenu, mjestu i uzroku smrti. Dodatno, oni mogu poslužiti kao pouzdani zamjenski uzorci za toksikološke analize u slučajevima kada ljudska tkiva i tekućine, koji se uobičajeno uzimaju tijekom obdukcije, ne postoje zbog raspadanja trupla. Ovaj rad prikazuje rezultate analize droga u uzorcima ličinki dviju porodica muha (Diptera: Calliphoridae i Sarcophagidae), prikupljenima s tijela muškarca srednje Životne dobi koji je počinio samoubojstvo otprilike 3 tjedna prije pronalaska njegova mrtvog tijela. Višestruki uzorci raspadajućeg ljudskog tkiva i ličinki muha analizirani su uporabom plinske kromatografije uz detekciju spektrometrijom masa (GC/MS) i u svima su otkriveni amfetamini. Dok su rezultati preliminarne kvalitativne analize bili izvan svake sumnje, kvantitativni su rezultati bili manje jasni te se preporučuje daljnje istraživanje u ovom području.

The Role of Alcohol in Road Traffic Accidents With Fatal Outcome: 10-Year Period in Croatia Split–Dalmatia County

Objective: The aim of the study was to investigate the relationship between alcohol consumption and fatal road traffic accidents that took place in Split–Dalmatia County (Croatia) during a 10-year period (from August 2001 to August 2011). Methods: A retrospective analysis of 474 autopsy reports was performed, with an emphasis on 337 toxicologal findings of blood alcohol concentration (BAC) and the mode of participation in traffic (driver, passenger, pedestrian, or cyclist). Relations between the BAC, the weekday on which death occurred, victims age and gender, and differences according to 3 successive legislation periods within the observed time frame were analyzed. The BAC was measured by gas chromatography with headspace and flame ionization detection. Results: Alcohol was regarded as a contributive death factor if BAC was positive; that is, higher than 0.5 g/kg. BAC was positive in 177 cases. Most often it was the car driver who had a positive BAC. Victims of traffic accidents were mostly male drivers, and those accidents were more often associated with alcohol consumption. Consumption of alcohol produced a significant increase in culpability. Conclusion: The results of our study show that alcohol remains one of the main contributing factors of traffic accidents in Split–Dalmatia County. Unfortunately, legislation changes regarding the allowed BAC did not cause any reduction in casualties among drivers driving under the influence of alcohol (DUIA). Chi-square test of the number of total driver victims and driver victims who were DUIA revealed no significant difference (P = .173).

Blood alcohol stability in postmortem blood samples.

AbstractThe effect of long-term storage on alcohol stability in postmortem blood samples was investigated. Seventy-nine blood samples were used in this study. For the first time, blood alcohol concentrations were measured within 1 to 4 days after being taken at the Laboratory of Forensic Toxicology. After storage (ranging from 191 to 468 days), each sample was reanalyzed for blood alcohol concentration, and results were compared with the original analysis.Results showed good agreement between the experimental measurements. About 90% of the results lie within 95% limits, and 10% were outside. Despite these positive correlations, the results of some blood samples showed a high variation in ethanol concentration. Deviation in alcohol concentration between replicates of up to 10% (observed in 39% samples) is not acceptable as precise forensic evidence.

Fatal Poisoning By Alcohol and Heroin

Fatal Poisoning By Alcohol and Heroin Drug abuse with alcohol consumption have been on the rise in Split-Dalmatian County for a while now. This article reports two separate cases with three deaths due to fatal combinations of heroin and alcohol. The first case of poisoning is related to a young couple, a 30-year-old man and a 28-year-old woman, who were found dead in a car, surrounded by cans of a variety alcoholic drinks. Two needles were found beside the bodies as well. The victims were registered drug abusers who had been in withdrawal programs. The second case was a 29-year-old man who was found dead in a house. Three fresh injection marks were visible on his right arm, and two needles were near his body. He was not known as a drug addict, but he had tried to commit suicide recently. Carboxyhaemoglobin was found in blood samples of both victims from the first case. The concentration was 25 % and that could contribute to their death. In both described cases blood alcohol concentration was higher then 1.60 g kg-1. Toxicology tests were positive for heroin, meconin, acetaminophen, 6-acetylmorphine, codeine, noscapine and papaverine. Ethanol, being a respiratory depressant, combined with morphine drastically increases the risk of rapid death due to respiration failure. Kobna Intoksikacija Alkoholom I Heroinom U posljednje vrijeme u Splitsko-dalmatinskoj županiji konzumiranje opojnih droga uz alkohol je u porastu. Ovim radom prikazali smo dva slučaja s tri smrtno stradale osobe koje su bile kobne žrtve kombinacije alkohola i heroina. U prvom slučaju, mladi par, u dobi od 30 i 28 godina, pronađen je u automobilu okružen limenkama raznih alkoholnih pića. Dvije šprice nađene su kraj tijela. Žrtve su bili registrirani ovisnici i na programima odvikavanja u organiziranim tretmanima. Drugi slučaj bio je 29-godišnji muškarac pronađen mrtav u napuštenoj kući. Na njegovoj desnoj ruci primijećene su tri svježe ubodne rane, a dvije šprice pronađene su kraj tijela. Nije bio poznat kao ovisnik o drogama, ali je nedavno pokušao samoubojstvo. Karboksihemoglobin nađen je u uzorcima krvi obiju žrtava iz prvog slučaja. Koncentracija je bila 25 %, što je moglo pridonijeti njihovoj smrti. U oba opisana slučaja maseni udjel alkohola u krvi bilo je veći od 1.60 g kg-1. Toksikološke analize bile su pozitivne na heroin, mekonin, acetaminofen, monoacetil morfin, kodein, noskapin i papaverin. Alkohol, i sam kao depresor respiracije, zajedno s heroinskom smjesom značajno povisuje rizik od nagle smrti kao posljedice zatajenja disanja.

Croatia is a safe tourist destination - study of foreign citizen mortality in Splitsko-dalmatinska and Primorsko-goranska County during the period 2001-2010.

Aim To investigate the mortality rate of foreign citizens in Croatia. Methods Data were collected from the Departments for Forensic Pathology in Split and Rijeka, which are the autopsy centers of the counties with approximately 35% of total foreign visitors, as well as from the Croatian Central Bureau for Statistics for the period 2001-2010. The mortality rate (number of deaths of members of each nationality per 100 000 entrances ratio) and standardized mortality ratio (ratio between the observed and expected number of deaths) were calculated, and χ2 goodness of fit test was used for statistical analyses. Results There were 447 deaths (325 men, 72.7%) of foreign citizens (mortality rate of 0.0015%). A total of 207 deaths (46.3%) were by natural causes, more often among older people, and 240 deaths (53.7%) were injury deaths, more often among younger people, mostly by drowning or traffic-related (22.2% and 18.6% of all deaths, respectively). Most represented were citizens of German, Austrian, Czech, and Italian nationality, with 115 (25.7%), 59 (13.2%), 58 (13.0%), and 52 deaths (11.6%), respectively. Mortality rate by nationality showed no significant difference (P < 0.05). The standardized mortality ratio was lowest in Hungarian and Czech citizens (0.17) and highest in US citizens (0.35). Conclusions Croatia has low foreign citizens’ mortality rate and could be considered a safe tourist destination.

A case of synthetic cannabinoid poisoning in Croatia

Abstract The number of new psychoactive substances (NPS), synthetic cannabinoids (SCs) in particular, is growing constantly. Because of the insufficiently explored effects on consumer health, they have become a major problem in the emergency departments. They are difficult to identify, and there are no antidotes that could reverse their detrimental effects. We report a case of poisoning of a young man who used SCs. The patient was admitted to the emergency department of the Clinical Hospital Merkur, Zagreb (Croatia) after sniffing and smoking a herbal product bought on the street. He presented with severe cognitive difficulties and visible eye redness. Other symptoms included somnolence, disorientation, loss of coordination, unsteady gait, hyporeflexia, stiffness, cramps and cold limbs, blurred vision, teeth grinding, dry mouth, tinnitus, fear, suicidal thoughts, impaired focus, memory, and speech, sedation, fatigue, depression, thought blocking, and autistic behaviour. His skin was dry, and his mucosa dry and irritated. Herbal products “Rainbow Special” and “Luminated Aroma” used by the patient were qualitatively analysed with gas chromatography / mass spectrometry (GC/MS) after direct extraction with an organic solvent. Solid-phase extraction method was used to analyse serum and urine samples. Despite the negative findings of biological samples, mostly due to the limitations of GC/MS, the clinical picture infallibly pointed to the poisoning with SCs. This was confirmed by the findings of 5-fluoro AMB (methyl 2-(1-(5-fluoropentyl)-1H-indazole-3-carboxamido)-3-methylbutanoate) in the herbal products.