Changes in SARS-CoV-2 viral load and titers over time in SARS-CoV-2-infected human corpses.

High viral titers of infectious severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have been detected in human corpses long after death. However, little is known about the kinetics of infectious SARS-CoV-2 in corpses. In this case series study, we investigated the postmortem kinetics of infectious SARS-CoV-2 in human corpses by collecting nasopharyngeal swab samples at multiple time points from six SARS-CoV-2-infected patients after their death. SARS-CoV-2 RNA was detected by quantitative reverse transcription-polymerase chain reaction from nasopharyngeal swab samples collected from all six deceased patients. A viral culture showed the presence of infectious virus in one deceased patient up to 12 days after death. Notably, this patient had a shorter time from symptom onset to death than the other patients, and autopsy samples showed pathological findings consistent with viral replication in the upper respiratory tract. Therefore, this patient died during the viral shedding phase, and the amount of infectious virus in the corpse did not decrease over time up to the date of autopsy (12 days after death). The findings of this study indicate that the persistence of SARS-CoV-2 in corpses can vary among individuals and may be associated with the stage of the disease at the time of death. These important results complement many previously reported findings on the infectivity of SARS-CoV-2 at postmortem.

Class III Alcohol Dehydrogenase Plays a Key Role in the Onset of Alcohol-Related/-Associated Liver Disease as an S-Nitrosoglutathione Reductase in Mice.

Lipid accumulation in the liver due to chronic alcohol consumption (CAC) is crucial in the development of alcohol liver disease (ALD). It is promoted by the NADH/NAD ratio increase via alcohol dehydrogenase (ADH)-dependent alcohol metabolism and lipogenesis increase via peroxisome proliferator-activated receptor γ (PPARγ) in the liver. The transcriptional activity of PPARγ on lipogenic genes is inhibited by S-nitrosylation but activated by denitrosylation via S-nitrosoglutathione reductase (GSNOR), an enzyme identical to ADH3. Besides ADH1, ADH3 also participates in alcohol metabolism. Therefore, we investigated the specific contribution of ADH3 to ALD onset. ADH3-knockout (Adh3-/-) and wild-type (WT) mice were administered a 10% ethanol solution for 12 months. Adh3-/- exhibited no significant pathological changes in the liver, whereas WT exhibited marked hepatic lipid accumulation (p < 0.005) with increased serum transaminase levels. Adh3-/- exhibited no death during CAC, whereas WT exhibited a 40% death. Liver ADH3 mRNA levels were elevated by CAC in WT (p < 0.01). The alcohol elimination rate measured after injecting 4 g/kg ethanol was not significantly different between two strains, although the rate was increased in both strains by CAC. Thus, ADH3 plays a key role in the ALD onset, likely by acting as GSNOR.

COVID-19 Analysis in Tissue Samples Acquired by Minimally Invasive Autopsy in Out-of-Hospital Deaths with Postmortem Degeneration.

Minimally invasive autopsy (MIA) is an alternative to a full autopsy for the collection of tissue samples from patients' bodies using instruments such as a biopsy needle. MIA has been conducted in many cases of coronavirus disease 2019 (COVID-19) and has contributed to the elucidation of the disease pathogenesis. However, most cases analyzed are hospital deaths, and there are few reports on the application of MIA in out-of-hospital deaths with varying extents of post-mortem changes. In this study, MIA and autopsies were performed in 15 patients with COVID-19 2-30 days after death, including 11 out-of-hospital deaths. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genome detection by reverse transcriptase quantitative polymerase chain reaction using MIA samples was mostly consistent with autopsy samples, particularly lung tissue, even in out-of-hospital cases. MIA had high sensitivity and specificity (> 0.80). Histological examination of lung tissue obtained by MIA showed characteristics of COVID-19 pneumonia, with 91% agreement with autopsy samples, whereas localization of SARS-CoV-2 protein in lung tissue was indicated by immunohistochemistry, with 75% agreement. In conclusion, these results suggest that MIA is applicable to out-of-hospital deaths due to COVID-19 with various postmortem changes, especially when autopsies are not available.

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.

Improved accuracy in high-frequency AC transport measurements in pulsed high magnetic fields.

We show theoretically and experimentally that accurate transport measurements are possible even within the short time provided by pulsed magnetic fields. For this purpose, a new method has been devised, which removes the noise component of a specific frequency from the signal by taking a linear combination of the results of numerical phase detection using multiple integer periods. We also established a method to unambiguously determine the phase rotation angle in AC transport measurements using a frequency range of tens of kilohertz. We revealed that the dominant noise in low-frequency transport measurements in pulsed magnetic fields is the electromagnetic induction caused by mechanical vibrations of wire loops in inhomogeneous magnetic fields. These results strongly suggest that accurate transport measurements in short-pulsed magnets are possible when mechanical vibrations are well suppressed.

Effects of long- and short-term experiences on stress during identification works of dead bodies: Rapid stress level measurement using voice.

OBJECTIVE: The mental health issues of personnel dealing with the deceased at times of disasters is a problem and techniques are needed that allow for real-time, easy-to-use stress checks. We have studied techniques for measuring mental state using voice analysis which has the benefit of being non-invasive, easy-to-use, and can be performed in real-time. For this study, we used voice measurement to determine the stress experienced during body identification training workshops for dentists. We studied whether or not stress levels were affected by having previous experience with body identification either in actual disaster settings or during training. DESIGN: Since participants training using actual dead bodies in particular are expected to suffer higher stress exposure, we also assessed their mental state pre- and post-training using actual dead bodies. RESULTS: The results confirmed marked differences in the mental state between before and after training in participants without any actual experience, between participants who engaged in training using manikins before actual dead bodies and participants who did not. CONCLUSIONS: These results suggest that, in body identification training, the level of stress when coming into contact with dead bodies varies depending on participants' experience and the training sequence. Moreover, it is believed that voice-based stress assessment can be conducted in the limited time during training sessions and that it can be usefully implemented in actual disaster response settings.

Suitability Evaluation of LaNi5 as Hydrogen-Storage-Alloy Actuator by In-Situ Displacement Measurement during Hydrogen Pressure Change.

The swelling ability of LaNi5 for application to hydrogen-storage-alloy (HSA) actuator is discussed through the measurement of the swelling ratio in hydrogen. The HSA actuator is driven by hydrogen pressure change causing the swelling of HSA. LaNi5 is one of the candidate materials for HSA actuators as well as palladium. Some prototypes of HSA actuators using LaNi5 have been fabricated; however, the kinetic swelling ability of LaNi5 itself has been not investigated. In this paper, the authors investigated the static and kinetic swelling ability of LaNi5 powder under hydrogen atmosphere. The results showed that the swelling ratio increased by 0.12 at the phase transition pressure. Response time decreased with an increase in the charged pressure during absorption, while it remained constant during discharge. Reaction kinetics revealed that these swelling behaviors were explained by hydrogen absorption and lattice expansion. The swelling ability of LaNi5 was also compared with that of palladium. The results show that LaNi5 swells 1.8 times more than palladium under 0.5 MPa. LaNi5 is suitable for an actuator driven repeatedly under more than the phase transition pressure. Palladium can be used for one-way-operation actuator even under 0.1 MPa since its response time during the evacuation was much longer than during the pressurization.

DNA analysis of hairs in a suspected case of child abuse.

Hairs are often used for DNA analysis in criminal investigations. DNA analysis of hairs with root sheaths is easy in many cases, but analyses using only the shaft or tip of the hair are often difficult. Here we describe a suspected case of child abuse in which we were commissioned to perform DNA analysis. Among 100 hairs, PCR amplification was succeeded in 99 samples, and as a result of direct sequencing, mitochondrial DNA (mtDNA) of the 99 hairs were classified into 6 types. The most common type was the 8-base substitution type of 16,168T-16,172C-16,183C-16,189C-16,217C-16,249C-16,325C-16,390A, which was observed in 86 hairs. This corresponded to the type of the victim. Total 736 STRs (75.5%) in 975 loci of 65 hairs could be typed, and only an amelogenin locus was typed in another hair. All 15 loci were typed in 10 hairs. STR types of 65 (98.5%) in 66 hairs were consistent with that of the victim. From 10 naturally-shed hair of a person, only 37 STRs (30.8%) in 120 loci of 8 hairs were typed, and all 15 loci could not be typed in these hairs. This difference in success rates of STR analysis suggested strongly that the hairs in this case were not shed naturally but forcibly, and the relevance to child abuse was suspected.

Moist-condition Training for Cerebrovascular Anastomosis: A Practical Step after Mastering Basic Manipulations.

As cerebrovascular anastomosis is performed in moist conditions that may impede precise manipulations, surgeons must undergo extensive preoperative training. We developed a simple moist-condition training method. It involves placing a free-floating inner platform hosting an artery from a chicken wing in an outer container filled with tap water to just below the specimen. Trainees performed anastomosis under magnification. Training sessions mimicked difficulties encountered during operations such as poor visibility of the lumen and problems handling the sutures. A retrospective comparison of 100 wet- and 100 dry-condition training sessions for end-to-side anastomoses with 8 stitches showed that under moist condition the time required for the entire procedure was significantly longer (17.8 ± 2.1 vs. 15.3 ± 2.1 min, p < 0.01) and the incidence of wrong stitching was greater (0.38 vs. 0%, p = 0.04). In 8 cases after introducing moist-condition training, the time required in superficial temporal artery to middle cerebral artery bypass surgery was significantly shorter than 8 cases before introducing the training (32.3 ± 5.6 min vs. 48.3 ± 15.9 min, p = 0.01). Incidence of wrong stitches was less in cases after introducing moist-condition training (2.7 vs. 7.4%, p = 0.10). Those indicate that moist-condition training is a useful and practical step and a bridge between training for basic manipulations under dry conditions and actual surgery.

Hanging foot switch for bipolar forceps: a device for surgeons operating in the standing position: technical note.

For surgeons operating in the standing position, the manipulation of foot switches involves shifting of the weight to the pivoting leg and the possible loss of contact between the switch and the foot. We solved this problem by changing the position of the switch that operates bipolar forceps. Our novel device is made of aluminum plates. The base plate features a foot strap and a height-adjustable overhang over the switch-operating foot. A commercially-available disc type foot switch is attached to the underside of the overhang in upside-down position, so the switch is operable with the toe. To turn on the switch, the toe is flexed dorsally to push the switch pedal, so the action is limited to the part distal to the metatarsophalangeal joints. Our switch was used in more than 100 consecutive microsurgeries performed by surgeons operating in the standing position. The switch manipulation required no shifting of the weight and was easier and quicker than manipulation of conventionally-placed switches. The surgeons were able to change the foot position freely with the modified switch, thereby avoiding loss of contact with the switch. The modified switch placement reduced physical fatigue in the lower extremities, annoyance related to the manipulation of conventionally-placed switches, and increased the comfort of surgeons operating in the standing position.

Modified wound dissection preserving the greater occipital nerve in foramen magnum decompression: a technique to reduce postoperative pain.

Patients undergoing foramen magnum decompression for Chiari malformation may experience severe postoperative pain in the area innervated by the greater occipital nerve (GON). We developed a modified dissection to lessen this pain. A midline skin incision was extended 2 cm in a cephalad direction to the inion and the skin was minimally retracted. After exposing the occipital bone, the semispinalis capitis and the trapezius muscles were detached subperiosteally in a caudal-to-cephalad direction. Consequently, the muscles and skin containing the GON were retracted in a single layer. We retrospectively compared the intensity of postoperative pain recorded on the visual analogue scale (VAS) by patients who underwent decompression using our (group A, n = 5) and the conventional layer-by-layer dissection technique (group B, n = 5). The VAS scores were not different on the day of surgery, but subsequently they fell faster in group A. Group A patients received a mild analgesic for a short period. Group B patients required a stronger analgesic for prolonged periods. Postoperative GON numbness/tenderness was observed only in group B. With respect to most evaluation criteria, the difference between the two groups was significant. Our anatomically rational dissection that protects the GON results in less postoperative pain.

[Dural scoring: an assistance technique for primary dural closure at craniotomy].

OBJECTIVE: At craniotomy the dura shrinks due to the drying effect of illumination and air exposure, rendering primary closure of this tissue difficult. We have developed a new technique, "dural scoring", that facilitates primary dural closure. PATIENTS AND METHODS: We used this technique in adults who underwent craniotomy when we encountered difficulties with primary dural closure of openings less than 5 mm in width. We placed scores along the edge of the opening on the surface of the dura (periosteal dura), taking care not to perforate the deep layer (meningeal dura). The dura relieved of tension expanded enough to be closed with sutures. RESULTS: This technique was successful in achieving primary dural closure in 53 patients who primarily underwent small supratentorial craniotomies. There were no procedure-related complications, e. g. cerebrospinal fluid leakages. CONCLUSIONS: Dural scoring is simple, requires no special instrumentation, and facilitates primary dural closure.

Anchoring of methylmethacrylate filler to the calvarium--technical note.

Implanted methylmethacrylate may be unexpectedly displaced due to poor adherence to the bone. We developed a simple technique to fix the material plugging the burr holes for use primarily in cosmetically important areas. At the closure of craniotomy, 2-3 small drill holes are made at the rim of the craniotomy burr hole. To address cranial defects in the pterional region, small holes are placed on the bone surface around the key burr hole. The holes extend into the diploic layer and have no parallel relationship. After fixation of the bone flap, a methylmethacrylate filler mixture is manually plugged into the burr hole and pushed into the small holes, thereby forming horns for secure fixation. None among over 100 patients developed an objectionable bulge attributable to displacement of the filler. Our technique requires no special instruments or materials and decreases the risk of cosmetic problems.

Inclined foot switches for surgical microscopes: a comfortable design for seated surgeons: technical note.

During microneurosurgical procedures, surgeons sitting on a stool may experience muscular fatigue in the lower half of the body due to repeat manipulation of foot switches controlling the surgical microscope. The mechanisms contributing to this fatigue were examined and a method developed for fatigue reduction. The switch panel harboring the pedals was originally designed to be placed horizontally, but was inclined toward the surgeon with a sandbag placed under the panel. The modified panel was used in consecutive surgeries by 3 surgeons. Surface electromyographs of the bilateral gastrocnemius and gluteus maximus muscles were obtained during simulated activation of the switches on horizontal and inclined panels using the left foot. Inclined panels facilitated stepping on the pedals, required less muscle power, and lessened fatigue and hip pain. Electromyographs obtained during stepping on the switches on the horizontal panel showed contraction of the left gluteus maximus, and the right gastrocnemius and gluteus maximus, muscles that do not directly contribute to stepping, in addition to contraction of the left gastrocnemius, a main muscle in the stepping motion, i.e. flexion of the ankle. Electromyographs obtained during stepping on the switches on the inclined panel showed muscle contraction was limited to the left gastrocnemius with lower frequency and amplitude motor unit potentials compared to those elicited during use of the horizontal panel. Our simple modification facilitates operating microscope manipulation and reduces surgeon fatigue.

Repositioning of Cranial Bone Flaps Cut with a Diamond-Coated Threadwire Saw: 5-Year Experience with Cosmetic Cranioplasty without Fixation Devices.

Artificial fixation systems for cranial bone flaps have problems related to their materials and designs. We developed an alternative technique for supratentorial craniotomy that employs a diamond-coated threadwire saw (diamond T-saw), originally developed for spinal surgery, and reduces the bone gap for fitted bone flap fixation. The study subjects were 77 adults undergoing elective supratentorial craniotomy. After placing a burr hole at each corner of the craniotomy, we performed osteotomy between adjacent burr holes to approximately one-third of the length of the osteotomy with a craniotome; this leaves a bony bridge at each corner. The diamond T-saw was introduced between adjacent burr holes through the epidural space and a bridge was cut with reciprocating strokes. On closure, the bridge firmly supports the flap and only sutures are needed for fixation. Successful bone flap fixation was obtained in all followed-up cases. There were no technique-related complications such as dural laceration, flap displacement, or resorption. Our method is ideal for bone cuts in cosmetic cranioplasty; it is easy, safe, and inexpensive and avoids the need for flap fixation with artificial devices.

Depth-adjustable fixation of external ventricular drains to counteract obstruction in tight ventricles.

Tightness of the lateral ventricle may result in mechanical obstruction of an external ventricular drain (EVD). We propose a modified EVD fixation method that allows retraction of the EVD to reopen the drainage. We used this technique in patients requiring long-term EVD placement in the frontal horn who were expected to develop tightening of the ventricle. We placed a catheter fixation device consisting of a dialysis catheter with a catheter-holding wing and a fixture spring. The wing is placed on the EVD just distal to its exit and tied down, and the fixture spring is attached to the wing to secure the EVD. If EVD obstruction associated with tightening of the ventricle was suspected, we removed the spring and retracted the EVD to the depth required for cerebrospinal fluid drainage, then replaced the spring. Retraction by 5-12 mm (mean 8.7 mm) resulted in opening of 8 of the 10 obstructed EVD devices. We encountered no major procedure-related complications. This modified EVD fixation method facilitates depth adjustments for reopening the obstructed drain.

Development of newly designed VHF interferometer system for observing earthquake-related atmospheric anomalies.

Temporal correlation between atmospheric anomalies and earthquakes has recently been verified statistically through measuring VHF FM radio waves transmitted beyond the line-of-sight. In order to locate the sources of such atmospheric anomalies, we developed a VHF interferometer system (bistatic-radar type) capable of finding the arrival direction of FM radio waves scattered possibly by earthquake-related atmospheric anomalies. In general, frequency modulation of FM radio waves produces ambiguity of arrival direction. However, our system, employing high-sampling rates of the order of kHz, can precisely measure the arrival direction of FM radio waves by stacking received signals.

Improvement of determination technique for extracting centerline in coronary artery with calcification on CTA.

The present study proposes a method for improving an existing technique used for extracting the centerlines of coronary arteries with calcifications on CTA. Through the use of the improved centerline extraction method, CPR images could be depicted accurately. In the proposed method, two dynamic range compression algorithms using two ITC for blood vessel regions and calcification regions were employed. Two dynamic range compressed CTA images obtained from the two ITC were then subtracted. After that, automatic tracing to determine the centerline for creating CPR images was performed using 3D image processing equipment. The usefulness of our proposed method was confirmed by using simulated CTA of the coronary artery with calcification. We also applied the proposed method to CTA images of 39 branches in 18 cases, and to 48 segments. Our experimental results showed that automatic tracing software pre-installed in the workstation could accurately determine the centerline of the coronary artery without being affected by the existence of calcification. Furthermore, an experienced radiological technologist evaluated the CPR images obtained by the proposed method. The results showed that most created CPR images could be used as diagnostic images after a minor revision performed manually.

Efficient generation of dopamine neuron-like cells from skin-derived precursors with a synthetic peptide derived from von Hippel-Lindau protein.

Skin-derived precursors (SKPs) from mammalian dermis represent neural crest-related stem cells capable of differentiating into both neural and mesodermal progency. SKPs are of clinical interest because they serve as accessible autologous donor cells for neuronal repair for neuronal intractable diseases. However, little is known about the efficient generation of neurons from SKPs, and phenotypes of neurons generated from SKPs have been restricted. In addition, the neuronal repair using their generated neurons as donor cells has not been achieved. The von Hippel-Lindau protein (pVHL) is one of the proteins that play an important role during neuronal differentiation, and recently neuronal differentiation of neural progenitor cells by intracellular delivery of a synthetic VHL peptide derived from elongin BC-binding site has been demonstrated. In the present study, a synthetic VHL peptide derived from elongin BC-binding site was conjugated to the protein transduction domain (PTD) of HIV-TAT protein (TATVHL peptide) to facilitate entry into cells, and we demonstrate the efficient generation of cells with dopaminergic phenotype from SKPs with the intracellular delivery of TATVHL peptide, and characterized the generated cells. The TATVHL peptide-treated SKPs expressed neuronal marker proteins, particularly dopamine neuron markers, and also up-regulated mRNA levels of proneural basic helix-loop-helix factors. After the TATVHL peptide treatment, transplanted SKPs into Parkinson's disease (PD) model rats sufficiently differentiated into dopamine neuron-like cells in PD model rats, and partially but significantly corrected behavior of PD model rats. The generated dopamine neuron-like cells are expected to serve as donor cells for neuronal repair for PD.