Genomic analysis of SARS-CoV-2 in forensic autopsy cases of COVID-19.

Numerous genomic analyses of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have been conducted, highlighting its variations and lineage transitions. Despite the importance of forensic autopsy in investigating deaths due to coronavirus disease 2019 (COVID-19), including out-of-hospital deaths, viral genomic analysis has rarely been reported due in part to postmortem changes. In this study, various specimens were collected from 18 forensic autopsy cases with SARS-CoV-2 infection. Reverse-transcription quantitative polymerase chain reaction revealed the distribution of the virus in the body, primarily in the respiratory organs. Next-generation sequencing determined the complete genome sequences in 15 of the 18 cases, although some cases showed severe postmortem changes or degradation of tissue RNA. Intrahost genomic diversity of the virus was identified in one case of death due to COVID-19. The accumulation of single-nucleotide variations in the lung of the case suggested the intrahost evolution of SARS-CoV-2. Lung of the case showed diffuse alveolar damage histologically and positivity for SARS-CoV-2 by immunohistochemical analysis and in situ hybridization, indicating virus-associated pneumonia. This study provides insights into the feasibility of genomic analysis of SARS-CoV-2 in forensic autopsy cases and the potential for uncovering important information in COVID-19 deaths, including out-of-hospital deaths.

Age estimation by palatal suture using modified Kamijo's method.

The present study aimed to investigate the correlation between palatal suture obliteration and age in modern Japanese and to develop an age estimation equation by modifying Kamijo's (1949) method. The subjects were 195 Japanese skeletal remains (155 males and 40 females) whose age and sex were known. First, obliteration score (OS) was obtained by measuring palatal suture obliteration from photographic images taken at the time of forensic autopsy, and the correlation with age was examined; no significant correlation was found in females. Second, the palatal sutures were divided into 14 sections, and each section was scored from 0 to 4 points according to the degree of the suture obliteration. Suture scores (SS) were then calculated for each of the four sutures, and the sum of the 14 scores (TSS: total suture score) was used to perform regression analysis for age. For male and all subjects (male and female), age significantly increased (p < 0.001) according to increment of SSs for all sutures. TSS has the highest regression coefficient (r = 0.540), and the lowest standard error of estimation (13.54 years) for all of the patients. The intra- and inter-observer agreement scoring showed high reliability. Validation study using the formulae showed a high percentage of correct responses (80 %). In conclusion, age estimation regression formula by palatal suture using modified Kamijo's method was established for Japanese population, and the study showed the formula might be valid for age estimation.

Age estimation by evaluating median palatine suture closure using postmortem CT.

This study sought to develop an age-estimation formula to evaluate the extent of median palatine suture (MP) closure using postmortem computed tomographic (PMCT) images. The PMCT images of 634 Japanese subjects (mean age, 54.5 years; standard deviation [SD], 23.2 years) with known age and sex were examined. The degree of suture closure of the MP, anterior median palatine suture (AMP), and posterior median palatine suture (PMP) was measured and scored (suture closure score, SCS), and a single linear regression analysis was conducted with age at death. On the analysis, SCS of MP, AMP, and PMP showed a significant correlation with age (p < 0.001). The correlation coefficient of MP was higher (0.760, male; 0.803, female; and 0.779, total) than that of AMP (0.726, male; 0.745, female; and 0.735, total) or PMP (0.457, male; 0.630, female; and 0.549, total). The regression formula and standard error of estimation (SEE) of MP were calculated as Age = 100.95 × SCS + 20.51 (SEE 14.87 years) for male subjects, Age = 91.93 × SCS + 26.65 (SEE 14.12 years) for female subjects, and Age = 95.17 × SCS + 24.09 (SEE 14.59 years) for the total, respectively. In addition, another 50 Japanese subjects were randomly selected to validate the age-estimation formula. In this validation, the actual age of 36 subjects (72%) was within the estimated age ± SEE. This study showed that the age estimation formula using PMCT images of MPs was potentially useful for estimating the age of unidentified corpses.

Statistical analysis of biomechanical properties and size of the sternum and its fracture risk in a Japanese sample.

OBJECTIVE: Sternal fractures can have life-threatening complications. To understand chest injury mechanisms, sufficient data regarding the mechanical properties and structure of the sternum are required. The aim of this study was to examine the mechanical properties and size of the sternum in a Japanese forensic sample. MATERIALS AND METHODS: Sterna were obtained from 120 Japanese dead bodies of known age and sex. The sample thickness (ST) and the sample width (SW) were measured using a computed tomographic image. Three-point-bending tests were conducted using a three-point-bending apparatus to assess the fracture load (FL) of the sample. Then, the flexural strength (FS) was calculated and the natural logarithm of FL (ln FL) and FS (ln FS) were also calculated. RESULTS: The values of ST, ln FL, and ln FS for male samples were significantly greater than those for female samples. Both ln FL and ln FS had significant negative correlations with age regardless of sex; the Pearson product-moment correlation coefficients were larger for female samples than for male samples. Although age was significantly negatively correlated with SW in female samples, there was no significant correlation between age and SW in male samples. No significant correlations were found between age and ST regardless of sex. CONCLUSION: This is the first study to present quantitative data on the biomechanical properties of the sternum. Because of the smaller sternal strength of elderly women, it is especially important for them to avoid the risk of sternal fractures.

Statistical analysis of the thickness and biomechanical properties of Japanese children's skulls.

OBJECTIVE: The structure and strength of a child's skull are important in accurately determining what and how external forces were applied when examining head injuries. The aims of this study were to measure skull thickness and strength in children, evaluate sex differences, and investigate the correlation between skull thickness and strength and age. MATERIALS AND METHODS: Skulls were obtained from 42 Japanese dead bodies under 20 years of age. During the autopsies, bone samples were taken from each skull. The length, width, and central thickness of the skulls were measured using calipers. Three-point bending tests were conducted, and bending load and displacement were recorded. Bending stress and bending strain were calculated, and Young's modulus, 0.2% proof stress, and maximum stress were obtained. RESULTS: In cases under 1.5 years old, 14 out of 46 male samples and 20 out of 40 female samples did not fracture during the three-point bending test, though no significant sex differences were detected. No significant differences in age, sample thickness, Young's modulus, 0.2% proof stress, or maximum stress were detected between the sexes. The sample thickness, Young's modulus, 0.2% proof stress, and maximum stress increased significantly and logarithmically with age (R2 = 0.761-0.899). Although age correlated with thickness, Young's modulus, and maximum stress more in females than in males, 0.2% proof stress correlated slightly better in males than in females. CONCLUSION: The skulls of preschool children, in particular, are thin, have low strength, and are at high risk of fracturing even with relatively small external forces. Unlike adults, no significant sex differences in skull thickness or strength were observed in children.

High titers of infectious SARS-CoV-2 in corpses of patients with COVID-19.

OBJECTIVES: The prolonged presence of infectious SARS-CoV-2 in deceased patients with COVID-19 has been reported. However, infectious virus titers have not been determined. Such information is important for public health, death investigation, and handling corpses. The aim of this study was to assess the level of SARS-CoV-2 infectivity in the corpses of patients with COVID-19. METHODS: We collected 11 nasopharyngeal swabs and 19 lung tissue specimens from 11 autopsy cases with COVID-19 in 2021. We then investigated the viral genomic copy number by real-time reverse transcription-polymerase chain reaction and infectious titers by cell culture and virus isolation. RESULTS: Infectious virus was present in six of 11 (55%) cases, four of 11 (36%) nasopharyngeal swabs, and nine of 19 (47%) lung specimens. The virus titers ranged from 6.00E + 01 plaque-forming units/ml to 2.09E + 06 plaque-forming units/g. In all cases in which an infectious virus was found, the time from death to discovery was within 1 day and the longest postmortem interval was 13 days. CONCLUSION: The corpses of patients with COVID-19 may have high titers of infectious virus after a long postmortem interval (up to 13 days). Therefore, appropriate infection control measures must be taken when handling corpses.

Identification, measurement, and evaluation of blood concentrations of insulin glargine and insulin lispro by UPLC-MS-MS in a dead body suspected of insulin overdose.

Insulin preparations, which are drug treatments for diabetes, cause fatal hypoglycemia when an overdose is administered. Cases of homicide and suicide using these preparations have been reported and are of great forensic interest. However, there are few reports assessing the postmortem concentration of insulin preparations, and it is often difficult to determine the cause of death. In the present study, we report a case of a suspected insulin glargine and insulin lispro overdose for suicide. A woman in her 30s had a history of mental illness and diabetes. The day before her death, she reported to her boyfriend that she had taken large doses of insulin preparations and prescription drugs. An autopsy revealed no fatal injuries or lesions. Drug screening tests revealed several prescription drugs, none of which showed toxic concentrations. Analysis using LC-MS/MS detected insulin glargine in the peripheral and cardiac blood at 429 µU/mL and 1362 µU/mL, respectively, whereas insulin lispro was detected in both the peripheral and cardiac blood at levels below the lower limit of quantification (LLOQ; <50 µU/mL). The cause of death was considered likely to be hypoglycemia caused by an overdose of insulin glargine. Insulin glargine is rapidly metabolized after subcutaneous administration and is rarely detected in the blood when used at therapeutic doses. There are no other reports on the quantification of insulin glargine parent compounds in postmortem samples, and this case provides important data on postmortem blood concentrations of insulin glargine intoxication.

Effect of ethanol on the diatom test using nitric acid or sodium hypochlorite.

OBJECTIVE: To study the quantitative effect of ethanol on the diatom test for water and lung samples. MATERIALS AND METHODS: In experiment 1, we tested 20 water samples taken from natural water areas. In experiments 1-1 and 1-2, each sample was digested with sodium hypochlorite (NaClO) solution (Purelox) and fuming nitric acid (HNO3), respectively. In Experiments 1-3 and 1-4, each sample was added to a lung sample containing few diatoms and digested with NaClO and HNO3, respectively. In Experiment 2, eight lung samples containing diatoms were digested with NaClO. Then, each digested sample was divided into two portions; one portion was washed with ethanol before being washed with water, and the other was washed only with water. After base-2 logarithmic transformation, the counts from the methods with and without ethanol wash were compared with a paired t-test. RESULTS: In experiments 1-1, 1-2, 1-3, 1-4, and 2, the geometric means of the ratios derived from the two methods (with/without ethanol) were 0.70 (95 % confidence interval [CI]: 0.65-0.77, P < 0.001), 0.83 (95 % CI: 0.73-0.93, P = 0.005), 3.00 (95 % CI: 2.31-3.91, P < 0.001), 0.91 (95 % CI: 0.79-1.04, P = 0.164), and 3.06 (95 % CI: 2.28-4.41, P < 0.001), respectively. CONCLUSION: Our experiments suggest that ethanol would be useless in diatom tests of water samples or in the conventional (HNO3) digestion of lung samples. However, ethanol is essential for the NaClO digestion of lung samples and could also be useful for other alternative methods of lung samples.

Detection of lisdexamfetamine and its metabolite d-amphetamine in urine and gastric contents collected from a cadaver at forensic autopsy.

PURPOSE: Lisdexamfetamine (LDX), which is used for the treatment of attention-deficit/hyperactivity disorder and narcolepsy, is composed of L-lysine attached to dextroamphetamine (d-amphetamine). In this article, we report a forensic autopsy case in which prescription drugs were unknown at autopsy. While amphetamine was detected, methamphetamine could not be detected by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in any of samples collected. Thus, we aimed to quantify LDX concentrations in autopsy samples and to prove that the amphetamine detected in this case was due to metabolized LDX. METHODS: Femoral vein blood, cardiac whole blood, urine, and gastric content samples were taken at autopsy for toxicological analysis. Qualitative and quantitative analyses were performed using LC-MS/MS. In addition, optical isomer separation for the amphetamine detected was conducted. The stability of LDX in whole blood and urine was also examined at three different temperatures. RESULTS: The concentrations of LDX were < 4.00, 30.9, and 4.42 ng/mL in whole blood, urine, and gastric content samples, respectively. The concentrations of amphetamine were 329, 510, 2970, and 915 ng/mL in femoral vein blood, heart whole blood, urine, and gastric contents, respectively. The amphetamine detected in this case was identified to be only d-amphetamine by optical isomer separation. The d-amphetamine detected was considered to be derived from LDX. Stability experiments revealed that LDX in whole blood decreased at ambient temperature. CONCLUSIONS: The results in the present case report may be useful in interpreting whether or not the amphetamine detected in a cadaver is a metabolite of LDX.

Methamphetamine-related forensic autopsy cases in a Japanese prefecture over a 7-year period: Characteristics of deaths and blood concentrations.

Understanding the actual conditions of methamphetamine (MA)-related death is important from the perspectives of criminal justice and public health. In this report, we review 104 cases of MA-related death handled by our departments between January 2014 and December 2020. Based on information from police and autopsy examinations, we classified the cases into the following categories: "accidental intoxication" ("MA only" and "multiple drugs or alcohol"), "fatal disease" ("definitively MA-related," "possibly MA-related," and "unlikely MA-related"), "accident," "suicide," "homicide," and "undetermined." The total number and annual trends for each category and their respective femoral blood concentrations were investigated. "Fatal disease" was the most common category (48 cases), followed by "suicide" (25 cases), "accidental intoxication" (14 cases), and "accident" (11 cases). "Definitively MA-related" in which MA may have played a role in their onset or exacerbation accounted for the majority of "fatal disease": 12 cases of heart disease, 4 cases of aortic dissection, 12 cases of cerebral hemorrhage, and 4 cases of subarachnoid hemorrhage. Cases classified as "definitively MA-related" died with lower femoral blood concentrations of MA compared with "MA only." Cases with "fatal disease" might have been misdiagnosed as "death by natural causes" if a proper autopsy and toxicology examinations were not performed. In death investigations, it is necessary to keep in mind that there are some MA-related deaths, and efforts should be made to increase awareness about the risk of death in using this drug.

Stature estimation by semi-automatic measurements of 3D CT images of the femur.

Stature estimation is one of the most basic and important methods of personal identification. The long bones of the limbs provide the most accurate stature estimation, with the femur being one of the most useful. In all the previously reported methods of stature estimation using computed tomography (CT) images of the femur, laborious manual measurement was necessary. A semi-automatic bone measuring method can simplify this process, so we firstly reported a stature estimation process using semi-automatic bone measurement software equipped with artificial intelligence. Multiple measurements of femurs of adult Japanese cadavers were performed using automatic three-dimensional reconstructed CT images of femurs. After manually setting four points on the femur, an automatic measurement was acquired. The relationships between stature and five femoral measurements, with acceptable intraobserver and interobserver errors, were analyzed with single regression analysis using the standard error of the estimate (SEE) and the coefficient of determination (R2). The maximum length of the femur (MLF) provided the lowest SEE and the highest R2; the SEE and R2 in all cadavers, males and females, respectively, were 3.913 cm (R2 = 0.842), 3.664 cm (R2 = 0.705), and 3.456 cm (R2 = 0.686) for MLF on the right femur, and 3.837 cm (R2 = 0.848), 3.667 cm (R2 = 0.705), and 3.384 cm (R2 = 0.699) for MLF on the left femur. These results were non-inferior to those of previous reports regarding stature estimation using the MLF. Stature estimation with this simple and time-saving method would be useful in forensic medical practice.

Occupational accidental injury deaths in Tokyo and Chiba prefectures, Japan: A 10-year study (2011-2020) of forensic institute evaluations.

Occupational accidental injury deaths (OAIDs) are a major social problem, and the analysis of individual cases is important for developing injury prevention measures. In this study, OAIDs with autopsies performed at forensic facilities in the metropolitan area of Japan (Tokyo and Chiba prefectures) from 2011 to 2020 were reviewed. The epidemiological characteristics of these OAIDs (n = 136), which accounted for 13.5% of OAIDs reported in the region during the study period, were compared with those of non-occupational accidental injury deaths (non-OAID) cases (n = 3926). Among OAID cases, 134 (98.5%) were men and 13 (9.6%) were foreign-born workers, which was significantly more than in non-OAID cases (p < 0.001, respectively). OAIDs were most frequent in construction (39.0%) followed by the manufacturing category (21.3%). The percentage of OAIDs in workers aged 65 and over showed an increasing trend. Most accidents occurred just after the start of work or just before the workday ended, as well as during the peak months of the year. The most common type of accident was fall/crash from a height (25.0%), and the most common injury site was the chest; none of these cases were confirmed to have been wearing a safety belt properly. Among foreign-born workers, the most common type of accident was caught in/between. As the working population is expected to change in the future, and an increase in the number of older adults and foreign workers is expected, it is necessary to take preventive measures such as improving the work environment based on ergonomics and providing safety education.

Ropinirole involved in a fatal case: blood and urinary concentrations.

PURPOSE: Ropinirole is an antiparkinsonian  drug and has recently been suggested to be effective in amyotrophic lateral sclerosis. It is expected that ropinirole prescriptions will increase in the near future. However, the fatal concentration in blood is unclear at this time. Therefore, we report a fatal case involving ropinirole intoxication and discuss the fatal concentrations with reference to several autopsy cases involving ropinirole. METHODS: Ropinirole was quantified in femoral vein blood, cardiac blood, and urine from five autopsy cases in which ropinirole was detected by drug screening in our laboratory. One is a ropinirole intoxication case (this report) and the others  were non-intoxication cases. Their ropinirole concentrations were compared and discussed. RESULTS: The ropinirole concentration in this case was 100 ng/mL in femoral blood, 160 ng/mL in cardiac blood, and 1840 ng/mL in urine. The ropinirole concentrations in the four non-ropinirole poisoning cases were 7-35 ng/mL (mean: 24 ng/mL) in femoral blood, 13-100 ng/mL (mean: 60 ng/mL) in cardiac blood, and 140-1090 ng/mL (mean: 640 ng/mL) in urine. Cardiac/peripheral ratios were in the range of 1.6-2.1 (mean 1.8). CONCLUSIONS: There were no obvious signs of overdose, and the high cardiac/peripheral blood ratio suggested that postmortem redistribution may have occurred, but the  peripheral blood ropinirole concentration (100 ng/mL) was obviously higher than that reported in the previous fatal case of ropinirole poisoning (64 ng/mL). Based on these results, the cause of death in this case was considered to be shock and fatal arrhythmia due to ropinirole poisoning. This case provides important data on postmortem blood and urinary levels of ropinirole poisoning.

Validity of dental findings for identification by postmortem computed tomography.

Postmortem computed tomography (PMCT) images help identify individuals and extract information from corpses. PMCT may substitute for a standard examination when bodies are severely damaged or when resources are limited in a mass fatality incident. In such situations, the dental information revealed by PMCT has the potential to narrow down candidates for identification further. However, the validity of the dental findings obtained from PMCT images remains unclear. In this study, we investigated the validity of dental findings on PMCT images compared to regular dental examinations as the reference standard. We routinely collected PMCT images of 148 unidentified corpses and compared the dental findings of each tooth obtained from PMCT with those of a regular dental examination. The validity of dental findings of PMCT was measured by sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). We also evaluated the accuracy of dental findings per corpse using 32 teeth as the denominator. The dental findings with high sensitivity and high specificity were a present tooth (0.96 and 0.97), a missing tooth (0.97 and 0.97), pontic (0.92 and 1.00), a dental implant (1.00 and 1.00), and a root filling (0.94 and 0.99). The mean accuracy of the dental findings per corpse in PMCT was 95.6% (standard deviation: 6.9, minimum ≤ median ≤ maximum: 65.6 ≤ 100 ≤ 100). The number of corpses with 100% accuracy in the dental findings was 81 (54.7%). The information obtained in this study highlights the potential use of PMCT during human identification in several settings with limited resources, such as the number of specialists present and the condition of the corpses.

Carrion flies (Insecta: Diptera) found on human cadavers in Chiba prefecture, Honshu, Japan, with the first record of Fannia prisca from a human corpse.

Forensic entomology (FE) involves the collection and analysis of necrophagous insects and arthropods for postmortem interval estimation based on their growth and succession. In Japan, research on FE is limited, and the predominant species of necrophagous insects and their distribution have not been clarified. The present study aimed to clarify the actual situation of insects, targeting Diptera collected from human cadavers, in Chiba Prefecture. We targeted the cases for which specimens could be collected from forensic autopsies conducted at the Legal Medicine Department of Chiba University Graduate School of Medicine or from human bodies handled by the Chiba Police in 2019. We specifically chose cases from which adult insects could be bred. Fifty cases were applicable, and 47 cases were successful in rearing adult flies. Sixteen species of Diptera were identified: six species of Calliphoridae, four species of Sarcophagidae, and a few species of Fanniidae, Muscidae, Phoridae, Piophilidae, and Stratiomyidae. The most frequently observed species were Lucilia sericata (20 cases) and Chrysomya pinguis (18 cases). For the first time, globally, we recorded the presence of Fannia prisca in a human cadaver. In conclusion, this is the first report to reveal the presence of carrion flies in human death cases in Chiba Prefecture, Japan. Research in FE is important in the context of the Japanese natural and social environment since this can aid forensic investigations conducted by authorities.

Prevalence of blood-borne infections in forensic samples: Epidemiology in areas of Chiba, Japan.

OBJECTIVES: To statistically clarify the prevalence and risk factors of infections in forensic autopsy cases in Chiba Prefecture, Japan. The aim was to improve preventive measures against infection in forensic autopsies. METHODS: We retrospectively investigated the positive detection rates of five infections (hepatitis B, HBV; hepatitis C, HCV; human immunodeficiency virus, HIV; human T-lymphotropic virus, HTLV; Treponema pallidum, TP) using 1491 samples obtained in forensic autopsy at our facility from 2014 to 2018. In addition, risk factors related to infection such as methamphetamine and tattoos were analyzed. Pearson's chi-square test was used for statistical analysis, and the difference was judged to be significant at p < 0.05. RESULTS: Among our samples, 9.0% of cadavers tested positive for infection, and the prevalence rates for HBV, HCV, HIV, HTLV, and TP were 1.0%, 6.7%, 0.3%, 0.7%, and 1.1% respectively. Statistically, cadavers linked to information about methamphetamine use had a 7.2 times higher rate of infection, and those with tattoos had a 5.6 times higher rate of infection, with HCV being the predominant cause. CONCLUSIONS: To limit the risk of infection among autopsy workers, cadavers and samples should be handled on the presupposition that the bodies are at risk of infections. It is also important to obtain as much information as possible about the medical history and potential illegal drug use to help assess the risk of infection in a patient during forensic autopsy. We propose that all autopsy cases should be screened for infections whenever possible.

Trends and Characteristics in Health Care-related Deaths Investigated through Medico-legal Autopsies after System Changes in Japan.

In Japan, a new cause-of-death investigation system and related new laws were enacted in the mid-2010s. These laws provided for an autopsy system for non-criminal unnatural deaths and a medical accident investigation system outside the criminal justice process for health care-related deaths. We retrospectively explored changes in the number and characteristics of medico-legal autopsy cases of health care-related deaths in Chiba Prefecture, Japan, and examined trends over time during these reforms. We found that the percentage of forensic autopsies based on the Code of Criminal Procedure for health care-related deaths had decreased significantly. The number of autopsies of accidental and unintentional deaths in nursing homes, which are not covered by the newly established medical accident investigation system, has been increasing, reflecting the ageing of society. The trend toward decriminalisation of health care-related deaths was expected to contribute more to medical safety if the scope was expanded and a system for disclosure of autopsy information was established.

Electrolyte analysis of pleural effusion for discrimination between seawater and freshwater drowning in decomposed bodies.

OBJECTIVE: The diagnosis of drowning is an important issue in forensic investigations. Moreover, discriminating between seawater and freshwater drowning is crucial to identify where the drowning occurred. The present study aimed to investigate electrolyte concentrations in pleural fluid in decomposed bodies in late postmortem intervals and derive cut-off values for the diagnosis of seawater and freshwater drowning. STUDY DESIGN: Data were collected from 44 seawater drowning cases, 60 freshwater drowning cases, and 30 non-drowning cases with pleural effusion which served as controls. The levels of sodium ion (Na+), potassium ion (K+), and chloride ion (Cl-) of pleural fluid were measured, and two indices were calculated: summation of Na+ and K+ levels (SUM Na + K), and summation of Na+, K+, and Cl- levels (SUM Na + K + Cl). The means of the three ion concentrations and two indices significantly differed between the three groups (p < 0.0001). RESULTS: The receiver operating characteristic analysis revealed that the sensitivity and specificity were both 1.000 for SUM Na + K + Cl of 288.3 mEq/L between the seawater and control groups. The Na+ value of 109.0 mEq/L also had a high sensitivity of 0.977 and a specificity of 0.933 in the seawater and control groups. The sensitivity and specificity were 0.967 and 1.000, respectively, for SUM Na + K of 123.2 mEq/L between the freshwater and control groups. CONCLUSION: The electrolyte concentrations in pleural effusion may be useful for the diagnosis of drowning in decomposed bodies with a longer postmortem interval.

The effect of ethanol or long-time reaction on the diatom test in water samples using sodium hypochlorite.

BACKGROUND: The diatom test method using sodium hypochlorite (NaClO) was equivalent to the conventional method in water samples. However, the method using NaClO was inferior to the conventional method in lung samples, in which ethanol was used and the reaction with NaClO was longer compared with the method in water samples. Using water samples, we aimed to clarify whether these differences affect the diatom test result. MATERIALS AND METHODS: Thirteen water samples from natural water sources were each divided into four parts corresponding to four (2 × 2) digestion methods: 3 "digestion" vs. 1 "digestion" and with ethanol vs. without ethanol. After the base-2 logarithmic transformation, the diatom counts were analyzed using three-way analysis of variance (ANOVA); factor 1 was "digestion times," factor 2 was "ethanol," and factor 3 was "sample number," and the interaction between factors 1 and 2 was also analyzed. RESULTS: The geometric means of the diatoms from the 3 "digestion" with ethanol method, the 3 "digestion" without ethanol method, the 1 "digestion" with ethanol method, and the 1 "digestion" without ethanol method were 373.5, 551.8, 436.6, and 522.0, respectively. ANOVA showed a significant difference in factor 2 (P = 1.7 × 10-4). There was no significant difference in factor 1 (P = 0.46), and no significant interaction between factors 1 and 2 (P = 0.13). CONCLUSION: Ethanol may decrease the diatom count in the diatom test using NaClO. In contrast, the diatom frustules do not dissolve through three-times digestion using NaClO.

SmartAmp method can rapidly detect SARS-CoV-2 in dead bodies.

Rapid and accurate detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in dead bodies is essential to prevent infection among those working with dead bodies. This study focused on the Smart Amplification (SmartAmp) method, which has a short examination time (approximately an hour), is simple to perform, and demonstrates high specificity and sensitivity. This method has already been used for clinical specimens; however, its effectiveness in dead bodies has not been reported. This study examined the SmartAmp method using 11 autopsies or postmortem needle biopsies performed from January to May, 2021 (of these, five cases tested positive for SARS-CoV-2 by quantitative real-time polymerase chain reaction (qRT-PCR) and six cases tested negative). Swab samples were collected from the nasopharynx, oropharynx, or anus and the SmartAmp and qRT-PCR results were compared. For the nasopharynx and oropharynx samples, the same results were obtained for both methods in all cases; however, for the anal swabs, there was one case that was positive according to qRT-PCR but negative according to the SmartAmp method. The SmartAmp method may therefore be less sensitive than qRT-PCR and results may differ in specimens with a low viral load, such as anal swabs. However, in the nasopharynx and oropharynx specimens, which are normally used for testing, the results were the same using each method, suggesting that the SmartAmp method is useful in dead bodies. In the future, the SmartAmp method may be applied not only during autopsies, but also in various situations where dead bodies are handled.