Antimicrobial resistance patterns of Streptococcus uberis isolates from bovine milk in Chiba prefecture, Japan: association between multidrug resistance and clonal complex 996.

Streptococcus uberis is one of major pathogens causing bovine mastitis. However, there is poor information on antimicrobial resistance (AMR) among the Japanese isolates. To provide treatment information for the mastitis caused by S. uberis in Japan, we aimed to clarify AMR patterns of the isolates from bovine milk mainly in Chiba. AMR phenotyping/genotyping [blaZ-erm(A)-erm(B)-mef(A)-linB-lnuD-tet(M)-tet(O)-tet(K)-tet(L)-tet(S)] and multilocus sequence typing were performed to analyze relationships between AMR patterns and clonal complexes (CCs). Resistance to tetracycline-, macrolide-, and lincosamide-classes was mainly associated with possession of tet(O), tet(S), erm(B), linB, and lnuD genes. CC996 was significantly associated with multidrug resistance (P<0.0001). These findings will aid Chiba farm animal clinics in treating bovine mastitis.

Potential for personal identification using the volume of the mastoid air cells extracted from postmortem computed tomographic images.

This study revealed the usefulness of volumetric analysis of mastoid air cells (MACs) extracted from postmortem computed tomography (PMCT) images in characterizing individuals. To characterize deceased persons, the MACs volumes of 61 Japanese PMCT images were measured after thresholding in Hounsfield units and based on the number of voxels on the right and left sides and the voxel size for each person. The volume differences between the right and left MACs and sex were examined. Although there were no obvious volume differences between males and females, the order of sizes on the two sides varied for each person. Moreover, deceased persons could be roughly classified using the total volume of MACs. Deceased persons with similar total volumes could be distinguished further by comparing the ratio of volumes in bilateral MACs. Although the identification process is dependent on samples and different sizes of bilateral MACs, our pilot study indicated that 81.9% (50/61) of deceased persons could be distinguished. In conclusion, volumetric analysis of MACs measured using PMCT imaging has the potential to identify individuals and reduce the number of candidates.

Development of an artificial intelligence-based algorithm to classify images acquired with an intraoral scanner of individual molar teeth into three categories.

BACKGROUND: Forensic dentistry identifies deceased individuals by comparing postmortem dental charts, oral-cavity pictures and dental X-ray images with antemortem records. However, conventional forensic dentistry methods are time-consuming and thus unable to rapidly identify large numbers of victims following a large-scale disaster. OBJECTIVE: Our goal is to automate the dental filing process by using intraoral scanner images. In this study, we generated and evaluated an artificial intelligence-based algorithm that classified images of individual molar teeth into three categories: (1) full metallic crown (FMC); (2) partial metallic restoration (In); or (3) sound tooth, carious tooth or non-metallic restoration (CNMR). METHODS: A pre-trained model was created using oral-cavity pictures from patients. Then, the algorithm was generated through transfer learning and training with images acquired from cadavers by intraoral scanning. Cross-validation was performed to reduce bias. The ability of the model to classify molar teeth into the three categories (FMC, In or CNMR) was evaluated using four criteria: precision, recall, F-measure and overall accuracy. RESULTS: The average value (variance) was 0.952 (0.000140) for recall, 0.957 (0.0000614) for precision, 0.952 (0.000145) for F-measure, and 0.952 (0.000142) for overall accuracy when the algorithm was used to classify images of molar teeth acquired from cadavers by intraoral scanning. CONCLUSION: We have created an artificial intelligence-based algorithm that analyzes images acquired with an intraoral scanner and classifies molar teeth into one of three types (FMC, In or CNMR) based on the presence/absence of metallic restorations. Furthermore, the accuracy of the algorithm reached about 95%. This algorithm was constructed as a first step toward the development of an automated system that generates dental charts from images acquired by an intraoral scanner. The availability of such a system would greatly increase the efficiency of personal identification in the event of a major disaster.

Ectopic cycle length estimation from the quantified distribution patterns of ventricular bigeminy and trigeminy.

BACKGROUND: Ectopic cycle length (ECL) and the distribution patterns of ventricular bigeminy and trigeminy, expressed as their postextrasystolic intervals (PEIs) and interectopic intervals (IEIs), have been poorly pursued. OBJECTIVE: Based on modulation theory, we hypothesized that the PEIs of bigeminy and trigeminy determine their IEIs and ECL. METHODS: Ambulatory electrocardiograms of 1290 patients with ventricular premature complexes (≥3000/day) were studied. To quantify their distribution pattern on the PEI vs IEI curve (PIC), we introduced the following 2 ratios: PEI of trigeminy to PEI of bigeminy ratio (T/B-PEI) and IEI of trigeminy to IEI of bigeminy ratio (T/B-IEI). Distribution patterns were divided into 3 types by T/B-PEI: standard type (<0.90), intermediate type (between 0.90 and 1.20), and reverse type (>1.20). ECL was defined as the average of the bigeminy and trigeminy intervals in the standard type, and bigeminy intervals in the other 2 types. RESULTS: T/B-IEI disclosed significant linear relationship with T/B-PEI (P < .0001). ECLs were longest in the standard type (1905 ± 347 ms; n = 426), followed by the intermediate type (1520 ± 239 ms; n = 607) and reverse type (1317 ± 227 ms; n = 227) (P < .0001). Trigeminy PEI/ECL in the standard type (0.450 ± 0.074) was significantly shorter than that of the other 2 types (P < .0001). CONCLUSION: We confirmed that T/B-PEI determines T/B-IEI and ECL by discriminating the 3 distribution patterns. Among them, trigeminy PEI/ECL decided the 2 types of modulation by the first sinus QRS, starting at the early delay phase or the later acceleration phase.

Examination of the usefulness of next-generation sequencing in mixed DNA samples.

The identification of individuals from mixed DNA samples is an important application of DNA typing. Although the discriminatory power of DNA profiling has improved dramatically, a limiting factor is that individuals cannot be identified via short tandem repeat (STR) analysis. We used next-generation sequencing (NGS) to examine the mixed DNA samples. Our results showed that STR nucleotide sequences and single nucleotide polymorphisms (SNPs) analysis via NGS may enable the identification of each distinct subject from a DNA mixture containing DNA of the victim and suspect.

Correction to: A simple method for the automatic classification of body parts and detection of implanted metal using postmortem computed tomography scout view.

In the original publication of the article, the authors affiliations were incorrectly published. The corrected affiliations are given in this correction.

Circumstances and factors of sleep-related sudden infancy deaths in Japan.

BACKGROUND: Sudden unexpected death in infancy (SUDI) comprises both natural and unnatural causes of death. However, few epidemiological surveys have investigated SUDI in Japan. OBJECTIVE: This retrospective study was conducted to investigate the latest trends of circumstances and risk factors of SUDI cases in which collapse occurred during sleep. METHODS: Forensic pathology sections from eight universities participated in the selection of subjects from 2013 to 2018. Data obtained from the checklist form were analyzed based on information at postmortem. RESULTS: There were 259 SUDI cases consisting of 145 male infants and 114 female infants with a mean birth weight of 2888 ± 553 and 2750 ± 370 g, respectively. Deaths most frequently occurred among infants at 1 month of age (18%). According to population data as the control, the odds ratio (95% confidence interval) of mother's age ≤19 years was 11.1 (6.9-17.7) compared with ages 30-39. The odds ratio for the fourth- and later born infants was 5.2 (3.4-7.9) compared with the frequency of first-born infants. The most frequent time of day for discovery was between 7 and 8 o'clock, and the time difference from the last seen alive was a mean of 4.1 h. Co-sleeping was recorded for 61%, and the prone position was found for 40% of cases at discovery. Mother's smoking habit exhibited an odds ratio of 4.5 (2.9-5.8). CONCLUSION: This study confirmed the trends that have been observed for sudden infant death syndrome; particularly, very high odds ratios were evident for teenage mothers and later birth order in comparison with those in other developed countries. Neglect was suspected in some cases of the prolonged time to discovery of unreactive infants. To our knowledge, this is the first report of an extensive survey of SUDI during sleep in Japan.

A simple method for the automatic classification of body parts and detection of implanted metal using postmortem computed tomography scout view.

Information on medical devices embedded in the body is important in the identification of an unidentified body. Computed tomography (CT) is a powerful imaging modality; however, metallic artifacts deteriorate the image quality because of the reconstruction method. On the contrary, CT scout view is less affected by metallic artifacts compared to CT. It is a simple method to classify the body into three rough parts for postmortem CT (PMCT) scout view, and an algorithm used to detect the location of the implanted metal has been developed for personal identification in forensic pathology. Of the test images, 97% were correctly classified into the three body parts. The true-positive rate for detection of the implanted metal in the scout view was 96.5%. Therefore, our simple methods are applicable in PMCT scout views and would be particularly useful for forensic pathology.

The derivation of predictive equations to speculate the post-mortem interval using cases with over 20-mL pleural effusion: A preliminary study.

Often, pleural effusion is noted at autopsy when the cause of death is drowning or diseases such as heart, renal and liver failure. Several studies have established a correlation between the concentrations of electrolytes in pleural effusion and the post-mortem interval (PMI) or those concentrations and drowning site. The present study aims to investigate the relationship between the amount of pleural effusion, concentrations of electrolytes and total protein in pleural effusion, by integrated interpretation using various factors such as the deceased's gender, age, cause of death, drowning site, PMI and body temperature. We included 40 cadavers (26 male, 14 female) with >20-mL pleural effusion, which were categorised into four groups as follows: freshwater drowning; brackish water drowning; seawater drowning (drowning group); and not drowning. An equation derived to assess the lung weight revealed that the drowning site affected the lung weight. An equation for the amount of pleural effusion in the drowning group for the first time revealed that the amount of pleural effusion was directly proportional to the PMI. Using an equation to assess the PMI, we could estimate the PMI within 13.0-13.2 h in cases with >20-mL pleural effusion. Despite a small number of cases in the present study, we attained exciting results from the integrated statistical analysis.

Predictive equation for post-mortem interval using spectrophotometric values of post-mortem lividity: A pilot study.

The colour of post-mortem lividity and control skin of 86 cadavers was measured spectrophotometrically to obtain L* (value), a* (chroma) and b* (hue) values. In addition, left heart blood (n = 58), right heart blood (n = 57) and blood from the femoral vein (n = 21) were measured. Using these data, we analysed the relationship between post-mortem lividity, control skin and blood colours. L* of post-mortem lividity (L*p) and control skin (L*c) were strongly correlated ( r = 0.64). a* and b* of post-mortem lividity (a*p and b*p) significantly increased with an increasing post-mortem interval (PMI) but r2 values were low (0.11 and 0.070, respectively). Predictive equations for post-mortem lividity colour (L*p, a*p and b*p) were developed using control skin colour (L*c, a*c and b*c) and autopsy findings for the first time. The predictive equation for L*p explained almost 65% of the observed L*p. We created predictive equations for PMI with and without blood colour values, and the most accurate equation, which did not use blood colour values, made it possible to estimate PMI within ± 10.29 h. Further study of these equations will help us to understand the factors that affect post-mortem lividity colour and increase the accuracy of equations for predicting post-mortem lividity colour and PMI.

Predictive equation to estimate post-mortem interval using spectrophotometric blood-colour values.

Forensic pathologists use post-mortem phenomena to estimate the post-mortem interval (PMI). We have reported on the usefulness of post-mortem lividity spectrophotometric values to estimate PMIs. Here, we focused on blood colour, looking for associations between blood colour, age and PMI. We generated predictive equations for blood-colour values and the PMI. We included data from a total of 129 cadavers (84 males and 45 females). We measured the colour of 124 left ventricular blood ( L*l, a*l, b*l), 123 right ventricular blood ( L*r, a*r, b*r) and 57 femoral blood ( L*f, a*f, b*f) samples. We found no significant associations between blood colour and age or between blood colour and the PMI, but the values of a*l, b*l, a*r and b*r were significantly increased with increased age, and those of L*f, a*f and b*f were significantly decreased with increased PMI. We created equations to estimate blood colour. The equations for femoral blood colour had higher adjusted R2 values and lower root mean square error values than those for left and right ventricular blood colours. We generated equations to estimate PMIs using blood-colour values and autopsy findings. Our estimated PMIs up to 67 hours had accuracies within 8.84 hours, without measuring post-mortem lividity colour or considering the age of the deceased. This is the first study to estimate PMIs based on blood-colour spectrophotometric values.

Postmortem interval estimation using the animal model of postmortem gas volume changes.

It is important to estimate the postmortem interval in forensic autopsy. Many methods to estimate the postmortem interval have been reported, and are typically associated with internal examination. However, there are issues such as rejection of autopsy by the family and a lack of forensic doctor in internal examination. Therefore, it is necessary to develop new methods, such as autopsy imaging, that can substitute for internal examination. Here, we first evaluated whether gas volume in the body increased with postmortem interval. Time-dependent X-ray CT imaging of euthanized Crl:CD (SD) rats (n = 3) was performed immediately after euthanasia and at seven subsequent time points up to 168 h (7 days) at 24-hour intervals. The data revealed that gas volume in the body increased in a time-dependent manner. Next, we reconstructed 3D images of isolated gas and calculated the gas volume using Amira software. In all cases, the volume of both portal venous gas and intestinal gas increased in a time-dependent manner. The volume of portal venous gas increased exponentially, while the volume of intestinal gas increased in a linearly with time. These data might be suggested that the postmortem gas volume change is one of indicators for estimating the postmortem interval. In addition, it would be possible to estimate more accurate postmortem interval by combining not only gas volume changes at the above two sites but also gas volume changes of the other sites such as heart cavities, kidney parenchyma, or abdominal aorta.

Bone comparison identification method based on chest computed tomography imaging.

The aim of this study is to examine the usefulness of bone structure extracted data from chest computed tomography (CT) images for personal identification. Eighteen autopsied cases (12 male and 6 female) that had ante- and post-mortem (AM and PM) CT images were used in this study. The two-dimensional (2D) and three-dimensional (3D) bone images were extracted from the chest CT images via thresholding technique. The similarity between two thoracic bone images (consisting of vertebrae, ribs, and sternum) acquired from AMCT and PMCT images was calculated in terms of the normalized cross-correlation value (NCCV) in both 2D and 3D matchings. An AM case with the highest NCCV corresponding to a given PM case among all of the AM cases studied was regarded as same person. The accuracy of identification of the same person using our method was 100% (18/18) in both 2D and 3D matchings. The NCCVs for the same person tended to be significantly higher than the average of NCCVs for different people in both 2D and 3D matchings. The computation times of image similarity between the two images were less than one second and approximately 10min in 2D and 3D matching, respectively. Therefore, 2D matching especially for thoracic bones seems more advantageous than 3D matching with regard to computation time. We conclude that our proposed personal identification method using bone structure would be useful in forensic cases.

Sex determination based on a thoracic vertebra and ribs evaluation using clinical chest radiography.

Our aim was to investigate whether sex can be determined from a combination of geometric features obtained from the 10th thoracic vertebra, 6th rib, and 7th rib. Six hundred chest radiographs (300 males and 300 females) were randomly selected to include patients of six age groups (20s, 30s, 40s, 50s, 60s, and 70s). Each group included 100 images (50 males and 50 females). A total of 14 features, including 7 lengths, 5 indices for the vertebra, and 2 types of widths for ribs, were utilized and analyzed for sex determination. Dominant features contributing to sex determination were selected by stepwise discriminant analysis after checking the variance inflation factors for multicollinearity. The accuracy of sex determination using a combination of the vertebra and ribs was evaluated from the selected features by the stepwise discriminant analysis. The accuracies in each age group were also evaluated in this study. The accuracy of sex determination based on a combination of features of the vertebra and ribs was 88.8% (533/600). This performance was superior to that of the vertebra or ribs only. Moreover, sex determination of subjects in their 20s demonstrated the highest accuracy (96.0%, 96/100). The features selected in the stepwise discriminant analysis included some features in both the vertebra and ribs. These results indicate the usefulness of combined information obtained from the vertebra and ribs for sex determination. We conclude that a combination of geometric characteristics obtained from the vertebra and ribs could be useful for determining sex.

Analysis of postmortem changes in internal organs and gases using computed tomography data.

Postmortem computed tomography (PMCT) is a useful method to identify various causes of death and measure the volume of internal organs and gases. The purpose of this study was to investigate postmortem changes as measured by PMCT, and the relationship between the volume of organs and gases and postmortem interval (PMI). Forty-six cadavers (22 men, 24 women) were examined by CT before autopsy. The volumes of the lungs, intrahepatic gas, and intrarectal gas were measured by CT using a workstation. A stepwise regression analysis was used to establish a predictive equation to ascertain the measured volume using factors including sex, age, height, body mass index (BMI), body surface area (BSA), and PMI. For estimation of PMI, stepwise regression analysis was used. In the equations for each measured volume, height, diaphragmatic height, and BSA were adopted for the left lung; height and diaphragmatic height were adopted for the right lung; PMI was adopted for intrahepatic gas; and sex and PMI were adopted for intrarectal gas. In the PMI equations, left lung volume, intrahepatic gas, and intrarectal gas were adopted together with sex, weight, and BMI. Values of intrahepatic gas decreased with postmortem changes, while intrarectal gas increased. This may be useful in investigation of postmortem changes. It will be necessary to include other parts of the intestine and to analyze volume changes in gases from these parts after death.

The assessment of status of tobacco smoking among urban primary schoolchildren in Madagascar.

The prevalence of tobacco smoking among adolescents aged 13-15 years old in Madagascar was previously reported to be higher than the average in other African regions. A preventive approach is urgently needed to avoid the initiation of early tobacco smoking. Therefore, the aims of this study were to evaluate the status of tobacco smoking among primary schoolchildren in Madagascar and explore the factors associated with initiation of tobacco smoking in the young. This study was conducted in the Mahajanga region of Madagascar. Three primary schools in this region and children of both genders between the ages of nine to 12 years old were randomly selected and approached to participate in this study. A self-administered questionnaire modified from the Global Youth Tobacco Survey Core Questionnaire 2007 was used to assess the status of tobacco smoking among primary schoolchildren. A total of 150 schoolchildren (14.0%) declared that they had smoked tobacco, with 30% starting to smoke tobacco at the age of seven or younger. The prevalence of tobacco smoking was lower among schoolchildren with non-smoking parents (p < 0.001) or non-smoking close friends (p < 0.001). Furthermore, schoolchildren's antismoking intention (p < 0.001) and their knowledge about the harmfulness of tobacco (p = 0.009) had significant effects on the prevalence of tobacco smoking. The results of this study indicate that tobacco smoking among schoolchildren in Madagascar may be influenced by peers, or parents, as well as smoking intention and knowledge about the harmfulness of tobacco use.

Expression of mRNA in the frontal cortex and hypothalamus in a rat model of acute carbon dioxide poisoning.

Acute carbon dioxide (CO2) poisoning causes no specific features that are revealed upon autopsy, and the pathophysiological mechanism of this syndrome is unclear. To address this issue, in the present study, we exposed rats to CO2 concentrations ranging from 10% to 60% and determined the effects on mRNA expression. According to the results of Gene Ontology (GO) and cluster analyses of microarrays data, we selected the following genes for further analysis: alkylglycerone phosphate synthase (Agps), hypocretin (Hcrt), tyrosine hydroxylase (Th), heat shock protein beta 2 (Hspb2), and opioid receptor delta 1 (Oprd1) expressed in the frontal cortex and renin (Ren), pancreatic polypeptide (Ppy), corticotropin releasing hormone receptor 2 (Crhr2), carbonic anhydrase 1 (Car1), and hypocretin receptor 1 (Hcrtr1) expressed in the hypothalamus. We found significant differences between the expression levels of Agps and Hspb2 mRNAs in the frontal cortex and that of Ppy, Crhr2 mRNAs in the hypothalamus in the presence of high concentrations of CO2. Further investigation of these genes may clarify the pathophysiology of acute CO2 poisoning and facilitate the development of novel forensic tests that can diagnose the cause of death.

[An Autopsy Case of Abnormal Behaviour Induced by Zolpidem].

Zolpidem is a widely used ultrashort-acting non-benzodiazepine in clinical practice; compared with benzodiazepines, it does not have side effects such as daytime hangover, rebound insomnia, and development of tolerance. We report an autopsy case of abnormal behaviour induced by zolpidem. A man in his 60's had suffered from postherpetic neuralgia about 2 months ago and had been prescribed zolpidem for insomnia. According to his family, he had no memory of his actions such as striking a wall, taking his futon outside, and eating 5 times a day after he took zolpidem. Because his postherpetic neuralgia did not improve, he was hospitalized and treated with an epidural block. During hospitalization, he took off his clothes, removed the epidural block catheter by himself, and slept on others' beds. He disappeared from the hospital one day; the next day, he was found dead in a narrow water storage tank 10 km away from the hospital. He was thought to have driven a car by himself to reach the place. Forensic autopsy revealed that the cause of death was drowning. Zolpidem and several other drugs were detected by toxicological analysis of his blood; the concentrations of these drugs were within therapeutic range. There are several reports about somnambulism induced by zolpidem such as sleepwalking, sleep driving, and eating. Considering the strange episodes following zolpidem administration, his behaviour on the day of his death was considered abnormal behaviour induced by zolpidem.

A fatal case of poisoning related to new cathinone designer drugs, 4-methoxy PV8, PV9, and 4-methoxy PV9, and a dissociative agent, diphenidine.

A woman in her thirties was found dead on a bed. Considerable amounts of "aroma liquid" and "bath salt" products and hypnotic drug tablets were scattered beside the bed. Autopsy showed pulmonary congestion and edema. Gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) analyses of "aroma liquid" and "bath salt" products showed the presence of new cathinone designer drugs, 4-methoxy PV8 (4-methoxy PHPP), PV9 (α-POP), and 4-methoxy PV9 (4-methoxy α-POP), and a dissociative agent, diphenidine. Drug screening in stomach contents, blood and hydrolyzed urine of the woman by GC-MS and liquid chromatography-tandem mass spectrometry (LC-MS/MS) revealed the presence of the above 4 types of drugs and 3 types of benzodiazepines, triazolam, flunitrazepam, and nitrazepam, and their metabolites. The above 7 drugs and 3 benzodiazepine metabolites were simultaneously determined by LC-MS/MS after modified QuEChERS (Quick, Easy, Cheap, Effective, Rugged, Safe) extraction using diazepam-d5 as the internal standard. The concentrations of 4-methoxy PV8, PV9, 4-methoxy PV9, and diphenidine in the femoral blood were 2.69, 0.743, 0.261, and 1.38µg/ml, respectively, which were significantly higher than concentrations reported in previous cases. Alcohol concentration in the femoral blood was 1.52mg/ml. Based on the pathological and toxicological findings, the cause of death was determined to be 3 types of cathinone drugs, 4-methoxy PV8, PV9, and 4-methoxy PV9, and diphenidine poisoning under the influence of 3 benzodiazepines and alcohol.

Anomalous origin of the right coronary artery from the left coronary sinus with an intramural course: comparison between sudden-death and non-sudden-death cases.

BACKGROUND: The prognosis of patients in whom the right coronary artery (RCA) arises from the left coronary sinus (LCS) is unequal. An initial intramural course of the coronary artery within the aortic media is considered to cause myocardial ischemia in cases of coronary anomalies. METHODS: Clinicopathological findings in five autopsy cases where the RCA arose from the LCS with an intramural course were examined. Comparison between sudden cardiac death and noncardiac death was also performed. RESULTS: Two of five cases were sudden cardiac death, and the other three cases were noncardiac death. In one case of sudden death, the person collapsed during light exercise, and in the other case, the person was under the effect of methamphetamine. Both of the cases of sudden death showed an RCA-dominant pattern in distribution of the coronary artery, atherosclerotic narrowing of the RCA, and acute ischemic necrosis in the posterior basilar ventricular septum around the atrioventricular conduction system, which is considered to be the territory of the RCA. CONCLUSIONS: An intramural course within the aortic media may be an accelerating factor of decreased blood flow in cases with an origin of the RCA arising from the LCS because of compression from the aortic lumen. However, this finding may not be an independent predictor of pathological ischemia. Additional factors that diminish blood flow in the intramural segment may be required to cause significant myocardial ischemia. Additionally, inciting factors, which can increase blood pressure, may also play a role in causing symptomatic myocardial ischemia by initiating mechanical compression from the aorta to the intramural segment of the RCA.