Disputed case of homicide by smothering due to severe amitriptyline intoxication of the victim

We report a fatal case of a female for whom the forensic autopsy revealed injuries to the external respiratory orifices indicating smothering. Subsequent postmortem toxicological analysis confirmed heavy amitriptyline acute intoxication. The victim had serious psychological problems, was under long-term treatment with antidepressants and was a systematic alcohol abuser. Forensic autopsy determined damage to the external airways, along with multiple formal petechial hemorrhages (Tardieu) in various parts of the body. The presence of amitriptyline, nortriptyline and 10-hydroxynortriptyline was confirmed by GC–MS and quantified by HPLC in blood (7.0 μg/ml amitriptyline and 7.4 μg/ml nortriptyline). The cause of death was disputed between severe intoxication (poisoning or suicide attempt) and smothering due to controversial evidence.     

An autopsy case of spontaneous esophageal perforation (Boerhaave syndrome)

A 45-year-old male, an alcohol addict with asthma, was found dead in his home, after several days of continued drinking. A forensic autopsy was performed 3 days after the discovery of his death in order to specify the cause of death.A longitudinal perforation penetrating all layers of the esophagus measuring 1.8 cm was present on the left wall approximately 2.0 cm from the gastroesophageal junction. There were 1900 mL of greenish to brownish turbid liquid in the left pleural cavity and 150 mL of greenish viscous liquid in the stomach. Histopathologically, an infiltration of numerous neutrophils was evident in the submucosa layer, proper muscular layer, and serous membrane of the esophagus, corresponding to the esophageal laceration. The serum C-reactive protein (CRP) concentration was determined to be 3.1 mg/dL. The alcohol concentrations were determined to be 1.49 mg/g in the right cardiac blood, 1.31 mg/g in the left cardiac blood, and 2.48 mg/g in urine.Based upon the autopsy and histopathological findings, as well as the biochemical and toxicological analyses, we concluded that the cause of death was respiratory failure by pleural effusion, resulting from spontaneous esophageal perforation. This was the first report of a spontaneous esophageal perforation eventually causing respiratory failure.     

Postmortem virtual volumetry of the heart and lung in situ using CT data for investigating terminal cardiopulmonary pathophysiology in forensic autopsy

Postmortem CT (PM–CT) is useful to investigate the viscera in situ before opening the body cavity at autopsy. The present study investigated heart and lung volumes in situ with regard to the cause of death as possible indexes of terminal cardiopulmonary dysfunction by means of PM–CT data analysis of forensic autopsy cases within 3 days postmortem (n = 70). Estimated heart volume was larger in sudden cardiac death (SCD; n = 10) and fatal methamphetamine abuse (n = 5) than in other groups, including mechanical asphyxiation (n = 12), drowning (n = 11), acute alcohol/sedative–hypnotic intoxication (n = 8), fire fatality (n = 12), hyperthermia (heatstroke; n = 6) and fatal hypothermia (cold exposure; n = 6). Estimated combined lung volume was larger in drowning, smaller in fire fatality due to carbon monoxide intoxication and SCD, and intermediate in other groups. Volume ratio of the lung to heart was higher in drowning, lower in SCD, and intermediate or varied in other groups; high and low ratios can indicate predominant/antecedent pulmonary and cardiac dysfunctions, respectively. These findings provide quantitative data that are not available at conventional autopsy or by routine two-dimensional CT morphology to assess three-dimensional gross heart and lung morphologies for interpreting terminal cardiopulmonary pathophysiology, detecting significant difference between SCD and other causes of death, especially mechanical asphyxiation and drowning.     

An unexpected death due to massive ascites and a giant mucinous ovarian cystadenoma

A female in her thirties fell face down in her room. She was motionless when her sister found her. She was transported to the hospital by ambulance and was in a state of cardiopulmonary arrest on admission. She did not respond to resuscitation. Her abdomen had started to swell 3 years before her death. An autopsy was performed to clarify the decedent’s cause of death. She was 172 cm tall and weighed 146 kg. Her maximum abdominal girth was 172.1 cm. A subcutaneous hemorrhage measuring 4.5 cm in diameter was observed in the epigastric region. The abdominal cavity contained brownish ascites (54.1 L). The left ovary was markedly swollen, and the combined weight of the uterus and right ovary was more than 13.0 kg. A left ovarian tumor consisting of serous and mucinous cysts was detected. There were no metastatic lesions in the peritoneum or other organs. She might have suffered circulatory disturbance caused by the ascites and ovarian tumor. Moreover, being in a prone position would have resulted in an increase in intra-abdominal pressure, further exacerbating her circulatory problems. Therefore, her cause of death was considered to be circulatory failure caused by significant ascites and a large ovarian tumor.     

Sudden unexpected neonatal death due to late onset group B streptococcal sepsis—A case report

We report a case of sudden unexpected death due to late onset neonatal group B streptococcal sepsis. A male neonate weighing 2731 g was born at 35 week gestational age, and discharged at the age of 4 days after the birth. At 6 days after the discharge (10 days after the birth), because of consciousness loss and hypothermia, the neonate was conveyed to an emergency hospital, eventually followed by his death. Forensic autopsy revealed neither severe trauma nor cardiac anomaly. Both lungs were edematous. Histopathologically, a lot of bacterial clusters were found in the lungs and intracerebral vessels. Cerebrospinal fluid contained a lot of leukocytes. Streptococcus agalactiae was detected in the specimens from the feces and the blood. Collectively, we diagnosed that the cause of the neonate’s death was late onset group B streptococcal sepsis. In autopsy cases of neonates, careful macroscopic and microscopic observations and bacteriological/virological examination should be performed.     

Postmortem urinary catecholamine levels with regard to the cause of death

Previous studies suggested that serum catecholamines are useful for investigating stress responses in the death process. The present study analyzed postmortem urinary adrenaline (Ad), noradrenaline (Nad) and dopamine (DA) in serial forensic autopsy cases (n = 199: 154 males and 45 females; age >9 years; survival time <0.5–168 h; within 10 days postmortem) to investigate the differences among the causes of death with special regard to hyperthermia (heatstroke; n = 11) and hypothermia (cold exposure; n = 10); other cases included fatalities from injury (n = 47), mechanical asphyxiation (n = 18), drowning (n = 14), intoxication (n = 31), fire fatality (n = 33) and natural death (n = 35). Each catecholamine level in urine was independent of the age or gender of the subjects, postmortem interval over 10 days or survival time, and did not correlate with the blood level. Urinary Adr and Nad levels were similar to those of clinical serum reference ranges, while DA was higher in all cases. Adr and Nad were higher in blunt head injury, methamphetamine abuse, hypothermia (cold exposure) and hyperthermia (heat stroke), but were low in mechanical asphyxia, drowning, fire fatality, sedative-hypnotic intoxication and acute cardiac death. DA was higher in injury, drowning, fire fatality, methamphetamine abuse and acute cardiac death, but was lower in mechanical asphyxiation and sedative-hypnotic intoxication. These profiles were quite different from those of serum levels, involving a predominant increase of DA, and may be useful for differentiating hyperthermia (heatstroke) and hypothermia (cold exposure) from drowning, sedative-hypnotic intoxication and sudden cardiac death.     

Blood concentrations of clobazam and norclobazam in a lethal case involving clobazam,meprobamate and clorazepate

Clobazam is a benzodiazepine with anti-anxiety and anticonvulsant properties marketed in several countries. Norclobazam, a metabolite of clobazam, has similar pharmacological activity but weaker sedative and tranquilizing effect. The two drugs were detected by GC–MS and determined by HPLC-DAD in the samples from a postmortem case. The femoral blood concentrations of clobazam and norclobazam were 0.72 and 36 μg/mL, respectively. The concentration of the active norclobazam was very high. The sum of both clobazam and norclobazam blood concentration (36.72 μg/mL) was clearly toxic, but was not necessarily fatal. Other associated drugs concentrations were within their therapeutic ranges. Interactions due to drug association were discussed.     

Hair analysis of an unusual case of Chloroquine intoxication

A dead body of middle aged man was exhumed from 6.5 month earth-grave. Autopsy findings were non-specific as the body was completely putrefied. Deceased’s scalp hair and kidney was sent for toxicological analysis. Hair sample (50 mg) was incubated with 1 M NaOH (2 ml). Chloroquine was detected in hair and kidney during basic drug screen performed on GC/MS. For confirmation and quantitation, chloroquine was extracted using Hypersep verify CX SPE cartridges while mass detector was operated in SIM mode using the ions of m/z 245.0, 290.1, 319.0 for chloroquine while ions of m/z 260 and 455 were monitored for nalorphine (internal standard). Chloroquine was present in high concentration in hair (211 ng/mg) as well as in kidney (37.3 mg/kg). Moreover, chloroquine was not detected in the wash solvents, suggesting ingestion of the drug rather than an external contamination of hair. These findings strongly suggested the acute exposure of higher doses of chloroquine to the deceased before death.     

Quantitative analysis of 3,4-dimethylmethcathinone in blood and urine by liquid chromatography–tandem mass spectrometry in a fatal case

We report here the quantitative analysis of cathinone-type designer drug 3,4-dimethylmethcathinone (3,4-DMMC) in blood and urine using liquid chromatography–tandem mass spectrometry (LC–MS/MS) in a fatal case. Abuse of 3,4-DMMC is widespread and a global issue. However, to date, there have been no reports of 3,4-DMMC-related deaths. We encountered a death in which 3,4-DMMC was thought to play a causative role, and successfully identified this designer drug from biological samples by using LC–MS/MS and QuEChERS (quick, easy, cheap, effective, rugged and safe) extraction method. For standard samples, detection of 3,4-DMMC in human blood and urine samples in the calibration range (5–400 ng/mL) was successful with recoveries of 85.9−89.4% (blood) and 95.8−101% (urine), limits of detection of 1.03 (blood) and 1.37 ng/mL (urine) and limits of quantification of 5.00 (blood) and 5.38 ng/mL (urine). The concentrations of 3,4-DMMC in blood (external iliac vein) and urine in the case were 27 mg/L and 7.6 mg/L, respectively. Some metabolites, including 3,4-dimethylcathione (DMC) and β-ketone reduced metabolites (β-OH-DMMC and β-OH-DMC), were detected in both blood and urine.     

Ruthenium porphyrin-induced photodamage in bladder cancer cells

Photodynamic therapy (PDT) is a noninvasive treatment for solid malignant and flat tumors. Light activated sensitizers catalyze photochemical reactions that produce reactive oxygen species which can cause cancer cell death. In this work we investigated the photophysical properties of the photosensitizer ruthenium(II) porphyrin (RuP), along with its PDT efficiency onto rat bladder cancer cells (AY27). Optical spectroscopy verified that RuP is capable to activate singlet oxygen via blue and red absorption bands and inter system crossing (ISC) to the triplet state. In vitro experiments on AY27 indicated increased photo-toxicity of RuP (20 μM, 18 h incubation) after cell illumination (at 435 nm), as a function of blue light exposure. Cell survival fraction was significantly reduced to 14% after illumination of 20 μM RuP with 15.6 J/cm², whereas the “dark toxicity” of 20 μM RuP was 17%. Structural and morphological changes of cells were observed, due to RuP accumulation, as well as light-dependent cell death was recorded by confocal microscopy. Flow cytometry verified that PDT-RuP (50 μM) triggered significant photo-induced cellular destruction with a photoxicity of (93% ± 0.9%). Interestingly, the present investigation of RuP-PDT showed that the dominating mode of cell death is necrosis.RuP “dark toxicity” compared to the conventional chemotherapeutic drug cisplatin was higher, both evaluated by the MTT assay (24 h).In conclusion, the present investigation shows that RuP with or without photoactivation induces cell death of bladder cancer cells.     

Accidental death of elderly persons under the influence of chlorpheniramine

Older individuals are susceptible to accident, such as falls, some of which are fatal. In such cases, autopsies and toxicological analysis may be deemed unnecessary, especially if the critical injuries and manner of death can be determined conclusively based on information at the scene and an external investigation. Here, we report the results of two autopsies performed on elderly individuals who died accidentally under the influence of chlorpheniramine. These autopsies revealed valuable additional information.Case 1: A woman in her 70s, who was living alone, was found dead under the stairs in her house. She had no history of a condition that could have led to sudden death. The autopsy revealed a neck fracture, multiple rib fractures, and a coccyx fracture. The histopathological findings showed fat embolisms in numerous small vessels of the interalveolar septum. Toxicological analysis of blood samples revealed the presence of chlorpheniramine (0.41 μg/ml).Case 2: A woman in her 70s, who was living alone, was found dead in the bathtub in her house. There was no past medical history other than diabetes mellitus and vertigo. The autopsy revealed hyper-inflated lungs and brown–red fluids in the trachea, but there was no evidence of a pathology or injury that could have induced a loss of consciousness. Toxicological analysis of the fluids in the right thoracic cavity revealed the presence of chlorpheniramine (0.57 μg/ml).In both cases, re-examination of the scene after the autopsy revealed the presence of common cold medicine containing chlorpheniramine. The victim may have accidentally overdosed on common cold medicine. This overdose would have been compounded by anti-histamine-induced drowsiness. The present cases suggest that forensic pathologists should always notify physicians/pharmacists of findings pertaining to unexpected drug side effects. Such intervention would prevent many accidental deaths. In addition, each autopsy must be performed in conjunction with a detailed postmortem investigation. Such efforts would also increase the accuracy of the public health record’s mortality statistics.     

A rare autopsy case of traumatic rhabdomyolysis associated with intermittent assault

Traumatic rhabdomyolysis generally occurs after severe blunt trauma and is acute in onset, associated with severe disease, and potentially lethal. Accordingly, diagnosis of traumatic rhabdomyolysis in patients without massive subcutaneous or intramuscular hemorrhage is difficult, especially in the postmortem period, which is limited in terms of the availability of biochemical examination tools and accurate history of illness. To the best of our knowledge, there are no previous reports of death from traumatic rhabdomyolysis among individuals who did not pursue medical consultation. A previously healthy man in his early sixties had been punched and kicked several times in the previous 2 months, but he had not gone to a hospital. He suddenly lost consciousness at his workplace approximately 5 days after the most recent assault, and cardiopulmonary arrest occurred when the emergency service arrived. He died the same day, and a medicolegal autopsy was performed. Although several sites of minor subcutaneous and muscle hemorrhage were observed, the cause of death was unclear upon macroscopic assessment. Immunohistochemical staining revealed acute renal failure caused by rhabdomyolysis. We herein report a rare case of fatal traumatic rhabdomyolysis, seemingly associated with minor and apparently nonlethal muscle injury.     

Development of a dual test procedure for DNA typing and methamphetamine detection using a trace amount of stimulant-containing blood

Investigation of drug-related crimes, such as violation of the Stimulant Drug Control Law, requires identifying the used drug (mainly stimulant drugs, methamphetamine hydrochloride) from a drug solution and the DNA type of the drug user from a trace of blood left in the syringe used to inject the drug. In current standard test procedures, DNA typing and methamphetamine detection are performed as independent tests that use two separate portions of a precious sample. The sample can be entirely used up by either analysis. Therefore, we developed a new procedure involving partial lysis of a stimulant-containing blood sample followed by separation of the lysate into a precipitate for DNA typing and a liquid-phase fraction for methamphetamine detection. The method enables these two tests to be run in parallel using a single portion of sample. Samples were prepared by adding methamphetamine hydrochloride water solution to blood. Samples were lysed with Proteinase K in PBS at 56 °C for 20 min, cooled at −20 °C after adding methanol, and then centrifuged at 15,000 rpm. Based on the biopolymer-precipitating ability of alcohol, the precipitate was used for DNA typing and the liquid-phase fraction for methamphetamine detection. For DNA typing, the precipitate was dissolved and DNA was extracted, quantified, and subjected to STR analysis using the AmpFℓSTR® Identifiler® Plus PCR Amplification Kit. For methamphetamine detection, the liquid-phase fraction was evaporated with N₂ gas after adding 20 μL acetic acid and passed through an extraction column; the substances captured in the column were eluted with a solvent, derivatized, and quantitatively detected using gas chromatograph/mass spectrometry. This method was simple and could be completed in approximately 2 h. Both DNA typing and methamphetamine detection were possible, which suggests that this method may be valuable for use in criminal investigations.     

An autopsy case of acetyl fentanyl intoxication caused by insufflation of ‘designer drugs’

We present a fatal case of intoxication due to insufflation of acetyl fentanyl. His blood concentration of acetyl fentanyl was 270 ng/mL, and the manner of death was classified as an accident. This is the first report of an autopsy case of acetyl fentanyl delivered by insufflation, rather than intravenous administration. He had been snoring loudly for at least 12 h prior to death, and transport to a hospital during this time and treatment with naloxone may have saved his life. In this sense, it can be said that his death was preventable.This case reemphasizes the risk of death associated with drug overdose and the narrow range of acetyl fentanyl between the effective dose (ED50) and lethal dose (LD50). The case should also raise awareness among medical professionals of the effectiveness of naloxone and the need to establish a comprehensive system for toxicological analysis while keeping the possibility of use of ‘designer drugs’ in mind.     

Preliminary screening method for the determination of inorganic arsenic in urine

A simple and rapid method was developed for the routine determination and classification of inorganic arsenic based on its clinical and forensic properties. Inorganic arsenic was isolated from urine by using copper granules, which was then made to react with ammonium molybdate in order to detect its presence with the naked eye. Based on studies of extraction and reaction conditions, e.g., reaction temperature and time, a colorimetric screening method was established. The reaction mixture was measured by a spectrophotometer, and there was linearity from 0.05 to 2.0 μg/ml and the correlation coefficients of the calibration curves were greater than 0.99. The coefficients of intra-day variation at 0.2 and 2.0 μg/ml of inorganic arsenic in urine were 9.6 and 4.2%, respectively (n = 5). The minimum detectable level in urine is 0.03 μg/ml, and it is possible to detect the lowest level of poisoning according to the published reports. The proposed method was applied to a poisoning case wherein the patient ingested NEOARSEN BLACK® with alcohol, which contained 45% of arsenic trioxide. This method produced positive results in all the urine samples tested, and this method is useful for the screening of inorganic arsenic based on its clinical properties because it enables the detection of inorganic arsenic in urine without expensive equipment.     

Cardiothoracic ratio in postmortem chest radiography with regard to the cause of death

It is difficult to examine the intact in situ status of thoracic organs, including the heart and lungs, after opening the chest at autopsy. The present study investigated the pathological diagnostic significance of the cardiothoracic ratio (CTR) with regard to heart and lung weight in postmortem plain chest radiography. The pathological diagnostic significance of the CTR in postmortem plain chest radiography using serial forensic autopsy cases of adults (>19 years of age, n = 367, within 72 h postmortem) was retrospectively investigated. In natural deaths, CTR was larger for heart diseases, and was smaller for pulmonary infection and gastrointestinal bleeding, showing correlations to the heart weight except in cases of hemopericardium. In traumatic deaths, CTR was larger in cases of fire fatality and acute methamphetamine intoxication, and varied in cases of blunt injury, showing correlations to the heart weight. However, CTR was smaller for sharp instrument injury and drowning, independently of the heart weight. These findings suggest that postmortem CTR (median, 55.6%, measured using a mobile X-ray apparatus) primarily depends on the heart weight, but is substantially modified during the process of death: the CTR may be enlarged by cardiac dilatation due to terminal congestive heart failure, but may be reduced by inflated lungs in drowning or hypovolemia due to fatal hemorrhage. CTR showed a mild correlation to the right diaphragm level, which was also related to the cause of death, but was independent of the left diaphragm level. Plain chest radiographic findings may also be helpful in investigating the pathophysiology of death, and are to some extent comparable with clinical findings. This also suggests the potential usefulness of postmortem CT and MRI for analysis of terminal cardiac function.     

Photodynamic therapy in VEGF inhibition non-responders—Pharmacogenetic study in age-related macular degeneration assessed with swept-source optical coherence tomography

BackgroundTreatment of neovascular age-related macular degeneration (nAMD) remains a major challenge in ophthalmology. It is essential to determine which of VEGF inhibition non-responders can benefit from photodynamic therapy (PDT). As AMD is strongly related to gene polymorphisms, genetic factors can modify efficacy of treatment. Swept-source optical coherence tomography (SS-OCT) gives exceptional insight into the retina and choroid. SS-OCT usefulness needs to be evaluated in nAMD patients.MethodsProspective 6-month study included consecutive 110 patients (110 eyes) with predominantly classic neovascular AMD treated with photodynamic therapy. Only non-responders to anti-VEGF were included in the study. Greatest linear dimension (GLD) of the lesion, best corrected visual acuity (BCVA), central subfield macular thickness (CSMT) and central choroidal thickness were assessed and compared between CFH and ARMS2 genotype groups. Success rate was the main endpoint. It was defined as not active CNV in the center of the fovea and no worsening in BCVA. Multiple regression was used to assess gene polymorphisms influence on PDT results. Wilcoxon tests were performed to determine significance of changes from baseline values.ResultsFollowing genotype frequencies were obtained—CFH CC 35 patients (31.8%), CT 52 (47.3%), TT 23 (20.9%); ARMS2 TT 28 patients (25.4%), GT 43 (39.1%), GG 39 (35.4%) success rate in CC/CT/TT CFH and TT/GT/GG ARMS2 groups were as follows respectively: 22.9%, 28.8%, 30.4% and 28.6%, 25.6%, 28.2%. The differences were not significant with highest odds ratio TT vs. CC CFH 1.57 (95% CI 0.48–5.2, p = 0.4). Significant increase in GLD was observed only in CC CFH group. Overall mean following measured parameters were obtained at baseline/day 7/month 3/month 6 (significant changes from baseline are marked with asterisk): GLD—3825 ± 1301 μm/3901 ± 1579 μm/3861 ± 1463 μm/3925 ± 1523 μm; CSMT—405 ± 203 μm/434 ± 257 μm*/321 ± 163 μm*/295 ± 157* μm; CCT—235 ± 103 μm/278 ± 157* μm/211 ± 113 μm*/201 ± 107* μm; BCVA—49.3 ± 12.5/43.2 ± 14.2*/49.6 ± 11.6/48.7 ± 12.2 letters on ETDRS charts. In all patients classic component of the lesion was assessed with SS-OCT with no need to be reaffirmed in FA. Thus FA was used mainly for lesion size calculation.ConclusionsCommon genetic factors seem not to influence PDT effectiveness in VEGF inhibitors non-responders. SS-OCT is a valuable tool of nAMD monitoring, especially for choroid assessment. Deterioration of retinal structure and function is observed one week after PDT. It is related to increase in both retinal and choroidal thickness and is accompanied by mild temporary BCVA decrease.     

Efficacy of erythrosine and cyanidin-3-glucoside mediated photodynamic therapy on Porphyromonas gingivalis biofilms using green light laser

BackgroundThe purpose of this in vitro study was to evaluate the efficacy of erythrosine and cyanidin-3-glucoside as photosensitizers in PDT for the elimination of Porphyromonas gingivalis (P. gingivalis) biofilms.MethodsP. gingivalis biofilms were prepared from a chronic periodontitis subject. Erythrosine and cyanidin-3-glucoside were prepared and randomly allocated as follows: 110, 220, 330, and 440 μM erythrosine; 101, 202, 303, and 404 μM anthocyanin; and 440 μM erythrosine + 404 μM cyanidin-3-glucoside. There were 18 PDT experimental groups (non-irradiated/irradiated with a 532-nm green light diode laser at 1.29 J/cm² for 60 s). The 3 controls were grouped as follows: biofilms exposed to the photosensitizers alone, biofilms exposed to the laser alone, and biofilms exposed to 0.12% chlorhexidine. All sample groups were cultured at 1, 3 and 6 h after PDT and incubated in an anaerobic chamber at 37 °C for 4 days. The surviving fraction was calculated from the log₁₀ CFU/ml. The 330 and 440 μM erythrosine and the 440 μM erythrosine + 404 μM cyanidin-3-glucoside were mixed with spin traps (TEMPO, DMPO), and the electron spin resonance spectra were evaluated.ResultsThe log₁₀ CFU/ml measurements showed that the PDT groups with 330 μM or 440 μM erythrosine and 440 μM erythrosine + 404 μM cyanidin-3-glucoside had statistically significant differences from the other groups (one-way ANOVA and Bonferroni’s multiple comparison test, p- value ≤ 0.05).ConclusionsPDT using 330 μM erythrosine, 440 μM erythrosine or 440 μM erythrosine + 404 μM cyanidin-3-glucoside irradiated with the laser more effectively inhibited P. gingivalis in biofilms.     

The in vitro effect of antimicrobial photodynamic therapy with indocyanine green on Enterococcus faecalis: Influence of a washing vs non-washing procedure

IntroductionThe purpose of this study was to evaluate the in vitro effect of washing and non-washing of indocyanine green (ICG) as photosensitizer (PS) on bacterial count, biofilm formation, development and degradation of Enterococcus faecalis.MethodsThe anti-bacterial, anti-biofilm formation, anti-biofilm development and biofilm degradation of anti-microbial photodynamic therapy (aPDT) against E. faecalis was determined at concentrations of 3 to 2000 μg/mL of ICG, subject to 18 J/cm² dose of diode laser (808 nm) in washing and non-washing producers. Bacterial viability measurements and biofilm assays were evaluated by broth microdilution method and crystal violet assays, respectively.ResultsICG-mediated aPDT, using 25 to 2000 μg/mL and 50 to 2000 μg/mL showed significant reduction in E. faecalis growth when compared to the control in non-washing and washing producers, respectively (P < 0.05). Also, ICG-mediated aPDT showed a significantly inhibitory effect on biofilm formation of E. faecalis in concentration of 6 to 2000 μg/mL and 100 to 2000 μg/mL in non-washing and washing groups (P < 0.05). The biofilm development was inhibited by concentrations of 12 to 2000 μg/mL and 100 to 2000 μg/mL in non-washing and washing groups. The biofilm degradation increased from concentrations of 12 to 2000 μg/mL and 250 to 2000 μg/mL in non-washing and washing groups, respectively.ConclusionThis study shows that the application of ICG should be accompanied by laser irradiation without being washed out to achieve better result for bacterial count reduction and anti-biofilm effects.     

Potential use of pericardial cTnI,Mg2+ and Ca2+ in the forensic investigation of seawater drowning in Greece: An initial assessment

The investigation of drowning constitutes one of the biggest problems in forensic practice. Elevated cardiac troponin I (cTnI) levels in biological fluids have been associated with myocardial damage, whereas increased Mg²⁺ and Ca²⁺ levels were found in cases of seawater drowning. The aim of this study was to examine the diagnostic utility of postmortem determination of cTnI, Mg²⁺ and Ca²⁺ in the pericardial fluid, in differentiating between cases of seawater drowning related to myocardial injury and those brought about by other causes. This study included 76 cases selected during a 2-year period from medicolegal autopsies. The cases were divided into three groups, according to the cause of death established based on macroscopic and microscopic evidence. The groups were: 1) seawater drowning (n = 23), 2) seawater drowning with histological evidence of myocardial infarction (n = 28), and 3) myocardial infarction unrelated to drowning (n = 25). cTnI was determined with an enzyme immunoassay; Mg²⁺ and Ca²⁺ with standard colorimetric assays. Pericardial cTnI levels were significantly lower in group 1 compared to groups 2 and 3. In contrast, pericardial Mg²⁺ and Ca²⁺ levels were both significantly higher in groups 1 and 2 compared to group 3. Our results suggest that the postmortem determination of pericardial cTnI levels may be useful in detecting previous myocardial damage as a contributory factor in death from seawater drowning and provide independent confirmation of the usefulness of pericardial Mg²⁺ and Ca²⁺ levels for differentiating between seawater drowning and fatal acute myocardial injury unrelated to the former.