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Article ; Online: From suicide to accident - Case report - The significance of the medico-legal autopsies.

Slović, Živana / Todorović, Miloš / Andrić, Ivana / Čanović, Vanja / Mihajlović, Filip / Vitošević, Katarina / Todorović, Danijela

Legal medicine (Tokyo, Japan)

2024 Volume 67, Page(s) 102397

Abstract: Introduction: Suicide is the intentional and voluntary act of destroying one's own life, while an accident is an unintended event that involuntarily causes injury to one's health or destruction of life.: Case presentation: We report a case of a 54- ... ...

Abstract	Introduction: Suicide is the intentional and voluntary act of destroying one's own life, while an accident is an unintended event that involuntarily causes injury to one's health or destruction of life. Case presentation: We report a case of a 54-year-old male who was found in a forest in late November, approximately 500 m away from his car. He was positioned bent over a trunk of a tree with his head beneath the rest of his body. His pants were down to his knees, and there were soiled blades of grass and leaves on his body. Investigation of the case circumstances revealed that he had attempted suicide by carbon monoxide poisoning, using a hose connected to the exhaust pipe and running it through the window into the cabin. Window on the driver's side was broken with glass particles on the driver's seat. Wrappers from "Rivotril" tablets, a generic benzodiazepine, were also found in the car. Autopsy revealed the following: postmortem hypostasis was of a cherry red color and well pronounced on the upper part of the front of the body and face. Numerous bruises, contusions, and erosions were present all over the body. Frostbites were especially pronounced in the knees and elbows area. The synovial membranes were partially bloodstained and reddish in color. Opening the stomach revealed erosions of the gastric mucosa (Wischnewsky sign). Chemical toxicological analysis detected presence of benzodiazepines and carboxyhemoglobin (25%). Conclusion: Based on the autopsy findings, chemical toxicological analyses, and investigation of the case circumstances, it has been concluded that the death occurred due to the combined effects of hypothermia, postural asphyxia and carboxyhemoglobin and benzodiazepine intoxication. The manner of death in this case is a combination of accidental and suicidal, as the victim attempted suicide but ultimately died due to exposure to low external temperature.
MeSH term(s)	Male ; Humans ; Middle Aged ; Carboxyhemoglobin/analysis ; Autopsy ; Accidents ; Contusions ; Benzodiazepines
Chemical Substances	Carboxyhemoglobin (9061-29-4) ; Benzodiazepines (12794-10-4)
Language	English
Publishing date	2024-01-11
Publishing country	Ireland
Document type	Case Reports ; Journal Article
ZDB-ID	2019555-2
ISSN	1873-4162 ; 1344-6223
ISSN (online)	1873-4162
ISSN	1344-6223
DOI	10.1016/j.legalmed.2024.102397
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Article ; Online: Beyond the virus: Examining the impact of COVID-19 on violent fatalities.

Andrić, Ivana / Slović, Živana / Todorović, Miloš / Čanović, Vanja / Mihajlović, Filip / Todorović, Danijela / Vitošević, Katarina

Legal medicine (Tokyo, Japan)

2024 Volume 67, Page(s) 102392

Abstract: Introduction: The COVID-19 pandemic has had a significant impact on various aspects of society, including crime rates. In Serbia, it is important to examine the changes in violent deaths before and during the pandemic to inform public health and safety ... ...

Abstract	Introduction: The COVID-19 pandemic has had a significant impact on various aspects of society, including crime rates. In Serbia, it is important to examine the changes in violent deaths before and during the pandemic to inform public health and safety policies. Materials and methods: We conducted a retrospective, epidemiological, cross-sectional analytical study of medico-legal autopsies in the Department of Forensic Medicine and Toxicology at the University Clinical Center of Kragujevac. Our study sample comprises all forensic autopsy cases examined from January 2017 to December 2019 (151 cases), labeled as "Before," and from January 2020 to December 2022 (192 cases), labeled as "During" the pandemic period. Natural deaths, skeletal remains, and undetermined cases were excluded from our sample. Results: The data show an increase in the total number of incidents reported from 152 in the "Before" period to 191 in the "During" period. The proportion of incidents involving males remained relatively stable at around 70%, while the proportion of incidents involving females increased. There was no statistically significant change in the proportion of incidents classified as accidental, while the proportion of incidents classified as homicide and suicide increased. The results show a statistically significant association between gender and incident type for both the "Before" and "During" periods. Deaths due to domestic violence have increased by 22.2% during the pandemic, which is cause for concern. In terms of demographic characteristics, males and younger individuals were more likely to be victims of violent deaths both before and during the pandemic. Conclusions: The COVID-19 pandemic had a significant impact on violent deaths in the Šumadija region (Central Serbia), with an overall increase in the number of violent deaths and a major impact on deaths due to domestic violence. Policies to address domestic violence should be prioritized during the pandemic and beyond, and strategies should be developed to mitigate the effects of future pandemics or lockdowns.
MeSH term(s)	Male ; Female ; Humans ; Cross-Sectional Studies ; Pandemics ; Retrospective Studies ; Violence ; Cause of Death ; Population Surveillance ; COVID-19 ; Communicable Disease Control ; Homicide ; Suicide
Language	English
Publishing date	2024-01-07
Publishing country	Ireland
Document type	Journal Article
ZDB-ID	2019555-2
ISSN	1873-4162 ; 1344-6223
ISSN (online)	1873-4162
ISSN	1344-6223
DOI	10.1016/j.legalmed.2024.102392
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Article ; Online: The use of vitreous humour as a potential source of DNA for postmortem identification in forensic science.

Čanović, Vanja / Slović, Živana / Todorović, Danijela / Todorović, Miloš / Andrić, Ivana / Radojević, Dušan / Ilić, Milena / Vitošević, Katarina

Forensic science, medicine, and pathology

2024

Abstract: Vitreous humour (VH) is routinely collected for toxicological analyses. However, only a few studies have examined its possible use in genotyping. Because of its isolation from the rest of the body, later onset of postmortem changes, and availability, VH ... ...

Abstract	Vitreous humour (VH) is routinely collected for toxicological analyses. However, only a few studies have examined its possible use in genotyping. Because of its isolation from the rest of the body, later onset of postmortem changes, and availability, VH could be a potential source of DNA for postmortem identification and other molecular analyses in forensic genetics. During forensic autopsies, samples of different VH volumes (5 ml, 4 ml, 3 ml, 2 ml, 1 ml, and 0.5 ml) were taken from 66 cadavers 24-48 h after death. DNA was also isolated from the samples immediately after collection and after a specific time of storage (1, 2, and 6 months). DNA was isolated using phenol‒chloroform‒isoamyl alcohol (PCI), and the yield and purity of the obtained DNA were determined spectrophotometrically using a FastGene NanoView Photometer. The integrity of the isolated DNA molecule was determined by PCR amplification of the hTERT (113 bp) gene. The results showed that VH could be a reliable source of genetic material for forensic analysis, and the method used for DNA extraction was effective. The yield of the isolated DNA ranged from 6.20 to 609.5 ng/µl, and the purity of the samples was 1.24-2.34. The isolated DNA concentration and integrity depend on the sample volume, but the DNA purity does not. It is also shown that DNA can be extracted from VH samples that have been stored for up to 6 months at - 20 °C. Therefore, using VH can be a valuable material for DNA identification.
Language	English
Publishing date	2024-01-03
Publishing country	United States
Document type	Journal Article
ZDB-ID	2195904-3
ISSN	1556-2891 ; 1547-769X
ISSN (online)	1556-2891
ISSN	1547-769X
DOI	10.1007/s12024-023-00760-w
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Article ; Online: Formalin Fixation of Human Healthy Autopsied Tissues

Todorovic Danijela / Vitosevic Katarina / Todorovic Milos / Slovic Zivana

Serbian Journal of Experimental and Clinical Research, Vol 21, Iss 4, Pp 307-

The Influence of Type of Tissue, Temperature and Incubation Time on the Quality of Isolated DNA

2020 Volume 315

Abstract: Formalin fixation is a widely used method in histopathology that has certain limits. Formalin often leads to the degradation of DNA molecules in cancer tissues, which makes tissues unusable for molecular analysis. The other factors may also affect the ... ...

Abstract	Formalin fixation is a widely used method in histopathology that has certain limits. Formalin often leads to the degradation of DNA molecules in cancer tissues, which makes tissues unusable for molecular analysis. The other factors may also affect the quality of DNA isolated from fixed tissues. The aim of this study is to determine the impact of the incubation time and temperature on the quality of DNA molecules isolated from various healthy human tissues. The brain, lung and kidney tissues, excluded during the forensic autopsies of people who died of violent death, were fixed in phosphate-buffered formalin from 24h to two months. After the completion of the incubation period, the DNA was isolated using phenol-chloroform-isoamyl alcohol extraction method and the concentration and purity of the samples were determined spectrophotometrically. The degree of degradation of DNA was assessed by PCR reaction, by amplification of gene fragments which lengths were 150bp (GPD1) and 262bp (β-actin). The highest concentration, purity and preserved integrity of DNA were obtained from the brain samples. With prolonged tissue incubation times in formalin, the concentration and integrity of DNA decreased in all tissue samples, especially in the brain tissue, while the purity of DNA remained unchanged. Also, tissue fixation at +4°C contributed to a better quality of isolated DNA compared to DNA isolated from tissue fixed at room temperature. We can conclude that the type of human healthy tissue, temperature and the incubation time of formalin fixation have important influence on the concentration, purity and integrity of DNA during fixation of tissues excluded in the course of forensic autopsy.
Keywords	forensic autopsy ; formalin ; dna isolation ; pcr ; spectrophotometry ; Medicine ; R
Subject code	612
Language	English
Publishing date	2020-12-01T00:00:00Z
Publisher	Sciendo
Document type	Article ; Online
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Article ; Online: Forensic characteristics of chest injuries among subjects who died in road traffic accidents

Slović Živana S. / Vitošević Katarina / Todorović Danijela / Todorović Miloš

Vojnosanitetski Pregled, Vol 78, Iss 2, Pp 215-

2021 Volume 222

Abstract: Background/Aim. In road traffic accidents, chest injuries are a critical factor since death usually occurs as a result of injuries to the heart and lungs, flail chest, pneumothorax, exsanguination, etc. The aim of this study was to analyze the most ... ...

Abstract	Background/Aim. In road traffic accidents, chest injuries are a critical factor since death usually occurs as a result of injuries to the heart and lungs, flail chest, pneumothorax, exsanguination, etc. The aim of this study was to analyze the most vulnerable subjects in road accidents, as well as the most frequent and most significant types of chest injuries sustained by different categories of subjects, and to examine the relevance of certain injuries or factors to outliving the injuries. Methods. The autopsy, the retrospective, and the cross-sectional study were performed at the Clinical Centre of Kragujevac, Department of Forensic Medicine and Toxicology. The study included all participants in road traffic accidents on the territory of Kragujevac and the surrounding area who died from the injuries sustained from the accidents or due to complications of the injuries during the period from 2001 to 2016. The subjects were divided into the following groups: pedestrians, motor vehicle drivers, front-seat passengers, back-seat passengers, bicyclists, motorcyclists, and tractor drivers. The occurrence of injuries in three regions of the body (chest, head, and abdomen) was analyzed in all the subjects. Results. The study included 525 subjects who died due to the injuries sustained in traffic accidents, which makes up to 38.4% of the total number of 1,366 autopsy cases covered by the study period. The average age of the subjects was 52 ± 19 years. The study sample consisted of 391 (74.5%) men and 134 (25.5%) women. The most vulnerable subjects were pedestrians (220, i.e. 41.9%), followed by motor vehicle drivers ( 98, i .e. 1 8.7%), front-seat passengers (79, i.e. 15%), motorcyclists (39, i.e. 7.4%), bicyclists (38, i.e. 7.2%), back-seat passengers (29, i.e. 5.5%) and tractor drivers (22, i.e. 4.2%). Chest injuries were identified in 408 subjects (77.7%), while the most frequent type of in-jury was rib fracture, observed in two-thirds of the sample. Out of the total number, 291 (55.4%) subjects died at the scene of the accident or on their way to the hospital, while 234 (44.6%) of them outlived injuries for a certain period of time. Drivers exhibited the highest risk of dying at the scene, while bicyclists outlived their injuries more frequently. Conclusion. Chest injuries are very common in subjects who died at the scene of the road traffic accident.
Keywords	accidents ; traffic ; thoracic injuries ; autopsy ; cause of death ; Medicine (General) ; R5-920
Subject code	380
Language	English
Publishing date	2021-01-01T00:00:00Z
Publisher	Military Health Department, Ministry of Defance, Serbia
Document type	Article ; Online
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Article ; Online: The quality of DNA isolated from autopsy formalin-fixed and formalin-fixed paraffin-embedded tissues: study of 1662 samples.

Vitošević, Katarina / Todorović, Danijela / Slović, Živana / Varljen, Tatjana / Radaković, Ivana / Radojević, Dušan / Čanović, Vanja / Todorović, Miloš

Molecular biology reports

2023 Volume 50, Issue 8, Page(s) 6323–6336

Abstract: Background: There are enormous formalin-fixed paraffin-embedded tissue archives and a constantly growing number of methods for molecular analyses but, the isolation of DNA from this tissue is still challenging due to the damaging effect of formalin on ... ...

Abstract	Background: There are enormous formalin-fixed paraffin-embedded tissue archives and a constantly growing number of methods for molecular analyses but, the isolation of DNA from this tissue is still challenging due to the damaging effect of formalin on DNA. To determine the extent to which DNA purity, yield and integrity depend on the process of fixation in formalin, and to what extent on the process of tissue paraffin embedding, we compared the quality of DNA isolated from fixed tissues and DNA isolated from tissues embedded in paraffin blocks after fixation. Methods and results: Heart, liver and brain tissues obtained from healthy people who suddenly died a violent death were fixed in 10% buffered formalin as well as in 4% unbuffered formalin for 6 h, 1-7 days (every 24 h), 10, 14, 28 days and 2 months. Additionally, the same tissues were fixed in 4% unbuffered formalin embedded in a paraffin block and stored from a few months to 30 years. The yield and purity of the DNA samples isolated from these tissues were measured using spectrophotometry. PCR amplification of the hTERT gene was performed to evaluate the degree of DNA fragmentation. Although the purity of the DNA isolated from almost all tissue samples was satisfactory, the DNA yields changed significantly. There was a decrease in successful PCR amplification of the hTERT gene in DNA samples isolated from tissue fixed in buffered and unbuffered formalin for up to 2 months from 100% to 8.3%. Archiving the tissue in paraffin blocks for up to 30 years also impacts the integrity of DNA, so there was a decrease in PCR amplification of the hTERT gene from 91% success to 3%. Conclusion: The largest decrease in DNA yield was observed after tissue formalin fixation after 14 days of fixation in buffered and unbuffered formalin. DNA integrity depends on the time of tissue formalin fixation, especially after 6 days for tissue fixed in unbuffered formalin, while for tissue fixed in buffered formalin the time is prolonged up to 28 days. The age of paraffin blocks also impacted DNA integrity, after 1 year and 16 years of archiving the paraffin blocks of tissues, there was a decrease in the success of PCR amplification.
MeSH term(s)	Humans ; Infant ; Formaldehyde ; Autopsy ; Paraffin Embedding ; Paraffin ; DNA/genetics
Chemical Substances	Formaldehyde (1HG84L3525) ; Paraffin (8002-74-2) ; DNA (9007-49-2)
Language	English
Publishing date	2023-06-13
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	186544-4
ISSN	1573-4978 ; 0301-4851
ISSN (online)	1573-4978
ISSN	0301-4851
DOI	10.1007/s11033-023-08491-5
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Article ; Online: New ruthenium(II) complexes with quinone diimine and substituted bipyridine as inert ligands: synthesis, characterization, mechanism of action, DNA/HSA binding affinity and cytotoxic activity.

Međedović, Milica / Rilak Simović, Ana / Ćoćić, Dušan / Senft, Laura / Matić, Sanja / Todorović, Danijela / Popović, Suzana / Baskić, Dejan / Petrović, Biljana

Dalton transactions (Cambridge, England : 2003)

2023 Volume 52, Issue 5, Page(s) 1323–1344

Abstract: This paper presents the synthesis and structural characterization of a series of new ruthenium(II) complexes 1-7, with the general ... ...

Abstract	This paper presents the synthesis and structural characterization of a series of new ruthenium(II) complexes 1-7, with the general formula
MeSH term(s)	Humans ; Ruthenium/pharmacology ; Ruthenium/chemistry ; Ligands ; Molecular Docking Simulation ; Antineoplastic Agents/chemistry ; DNA/chemistry ; Quinones ; Adenocarcinoma ; Coordination Complexes/chemistry ; Cell Line, Tumor
Chemical Substances	Ruthenium (7UI0TKC3U5) ; Ligands ; Antineoplastic Agents ; DNA (9007-49-2) ; Quinones ; quinone (3T006GV98U) ; Coordination Complexes
Language	English
Publishing date	2023-01-31
Publishing country	England
Document type	Journal Article
ZDB-ID	1472887-4
ISSN	1477-9234 ; 1364-5447 ; 0300-9246 ; 1477-9226
ISSN (online)	1477-9234 ; 1364-5447
ISSN	0300-9246 ; 1477-9226
DOI	10.1039/d2dt02993f
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Article ; Online: The changes in tissue histomorphology and quality of DNA in healthy human autopsied tissues stored at -20 °C and -150 °C.

Radojevic, Dusan N / Todorovic, Milos S / Vitosevic, Katarina M / Canovic, Vanja / Pavicevic, Milena / Slovic, Zivana S / Andric, Ivana M / Matic, Stevan / Todorovic, Danijela V

Legal medicine (Tokyo, Japan)

2024 Volume 67, Page(s) 102396

Abstract: The aim of this study was to compare the changes in tissue histomorphology and DNA quality in six different healthy tissues (brain, heart, lung, liver, spleen and kidney) exempted during autopsy of healthy individuals and storage at -20 °C and -150 °C ... ...

Abstract	The aim of this study was to compare the changes in tissue histomorphology and DNA quality in six different healthy tissues (brain, heart, lung, liver, spleen and kidney) exempted during autopsy of healthy individuals and storage at -20 °C and -150 °C three month. Tissue samples were obtained, divided by tissue and temperature group, and for each sample, tissue histomorphology and DNA (isolated from all tissues in duplicated - 72 samples of DNA) quality were analysed. Morphology of tissue samples was studied using H&E staining. DNA was isolated using the phenol-chloroform-isoamyl alcohol method. To assess the concentration and purity of the DNA samples, we used a spectrophotometer to measure absorbance at wavelengths of 280 nm and 260 nm. The fragments of human telomerase reverse transcriptase (hTERT) gene were amplified from the DNA using PCR reaction and then visualised using the 2 % agarose gel. Samples stored at -150 °C sustained the highest degree of histomorphological damage, while samples stored at -20 °C were less degraded, compared to control. The liver samples stored at -20 °C had a mean DNA concentration (1030.4 ± 51.5 ng/μl) higher than the samples of liver tissue stored at -150 °C (497.4 ± 167.1 ng/μl) (p < 0.001). Other tissues did not have statistically significantly different DNA concentration at both temperatures. Liver samples at -20 °C had degraded DNA, showed as the absence of hTERT gene in most of samples. Other tissue samples in both temperature groups had unfragmented DNA. Storing tissue samples at -20 °C is not inferior in terms of DNA yield and integrity, and possibly superior for tissue histomorphology, comparing with samples stored at -150⁰C.
MeSH term(s)	Humans ; DNA/genetics ; Autopsy ; Temperature ; Spectrophotometry
Chemical Substances	DNA (9007-49-2)
Language	English
Publishing date	2024-01-13
Publishing country	Ireland
Document type	Journal Article
ZDB-ID	2019555-2
ISSN	1873-4162 ; 1344-6223
ISSN (online)	1873-4162
ISSN	1344-6223
DOI	10.1016/j.legalmed.2024.102396
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Article ; Online: Formalin fixation at low temperature provides better yield and integrity of DNA

Makulović Stefan / Vitošević Katarina / Slović Živana / Todorović Miloš / Todorović Danijela

Medicinski Podmladak, Vol 70, Iss 2, Pp 37-

2019 Volume 42

Abstract: Introduction: Fixation with formalin, a widely used procedure to preserve tissue samples, leads to damage of DNA through crosslinking activity. The factors that could influence the fixation and integrity of DNA may be numerous: incubation time, tissue ... ...

Abstract	Introduction: Fixation with formalin, a widely used procedure to preserve tissue samples, leads to damage of DNA through crosslinking activity. The factors that could influence the fixation and integrity of DNA may be numerous: incubation time, tissue type, concentration of formalin, temperature, pH and viscosity. Aim: The aim of this investigation was to examine the influence of incubation time and temperature of formalin fixation on the yield, purity and integrity of DNA isolated from healthy human heart myocardial tissue taken during medico-legal autopsy. Material and methods: Heart tissue samples were fixed in phosphate-buffered formalin at +4°C in the dark, as well as at room temperature in the presence of light. The DNA was isolated after one day, then successively every day during the first week, and then on the tenth day, and after two and four-week periods using phenol-chloroform-isoamyl alcohol extraction method. The absorbances were measured at 260 nm and 280 nm, which allows calculation of yield and purity of nucleic acid in the samples. The PCR amplification of two genes, GPDH (150 bp) and ß-actin (262 bp), were performed to evaluate the degree of DNA molecule fragmentation. Results: The highest yield, purity and preserved integrity of DNA were obtained from the samples fixed in formalin at +4°C in the darkness. In these samples, GPDH and ß-actin genes were amplified up to 14 days, unlike the samples that were fixed at the room temperature in which the ß-actin gene was amplified up to 5 days, while the GPDH gene fragment was successfully amplified up to 10 days of fixation. Conclusion: The temperature, presence of light and the incubation time of formalin fixation all have important influences on yield, purity and integrity of DNA during the fixation process.
Keywords	formalin ; pcr amplification ; autopsy ; dna degradation ; spectrophotometry ; Medicine ; R
Subject code	630
Language	English
Publishing date	2019-01-01T00:00:00Z
Publisher	University of Belgrade, Medical Faculty
Document type	Article ; Online
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Article ; Online: Apoptosis and cell cycle

Petrović Marija / Todorović Danijela

Racionalna Terapija, Vol 6, Iss 1, Pp 21-

2014 Volume 32

Abstract: Apoptosis, a form of programmed cell death, is used to eliminate individual cells surrounded by normal cell population. It is a controlled way of cell death in which the cell actively participates by conducting precise, gene-regulated program of self- ... ...

Abstract	Apoptosis, a form of programmed cell death, is used to eliminate individual cells surrounded by normal cell population. It is a controlled way of cell death in which the cell actively participates by conducting precise, gene-regulated program of self-destruction, that is, cell 'suicide.' Active synthesis of macromolecules is necessary during this process. Death of individual cells is necessary to maintain a balance in living systems, so the process of apoptosis is continuously present in the body, which allows normal development, tissue homeostasis, and many other physiological processes. The molecular mechanisms that regulate apoptosis are functionally linked to other cellular mechanisms, such as control of the cell cycle, cell proliferation and differentiation, genomic stability and cellular metabolism. Damage to the DNA molecule, caused both spontaneously and under the influence of various chemical and physical agents, leads to the cell cycle arrest and activation of mechanisms that repair the damage. Depending on the type and extent of the damage, the cell either continues progression through the cell cycle, or activates the mechanisms that lead to apoptosis. Disturbances in the regulation of apoptosis and cell cycle present the molecular and biological basis of many diseases. Because of the importance of these processes during the development and progression of tumors, their use as biological markers is one of the main strategies in the formation of therapeutic approaches for the treatment of cancer.
Keywords	apoptosis ; cell cycle ; cancer ; Medicine (General) ; R5-920
Subject code	610
Language	English
Publishing date	2014-01-01T00:00:00Z
Publisher	Medicinsko društvo za racionalnu terapiju Republike Srbije
Document type	Article ; Online
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