Characterization of SARS-CoV-2 and host entry factors distribution in a COVID-19 autopsy series.

Background: SARS-CoV-2 is a highly contagious virus that causes the disease COVID-19. We have recently reported that androgens regulate the expression of SARS-CoV-2 host entry factors ACE2 and TMPRSS2, and androgen receptor (AR) in lung epithelial cells. We also demonstrated that the transcriptional repression of the AR enhanceosome inhibited SARS-CoV-2 infection in vitro. Methods: To better understand the various sites of SARS-CoV-2 infection, and presence of host entry factors, we extensively characterized the tissue distribution and localization of SARS-CoV-2 virus, viral replication, and host entry factors in various anatomical sites sampled via autopsy. We applied RNA in-situ-hybridization (RNA-ISH), immunohistochemistry (IHC) and quantitative reverse transcription polymerase chain reaction (qRT-PCR) approaches. We also assessed histopathological changes in SARS-CoV-2 infected tissues. Results: We detect SARS-CoV-2 virus and viral replication in pulmonary tissues by RNA-ISH and IHC and a variety of non-pulmonary tissues including kidney, heart, liver, spleen, thyroid, lymph node, prostate, uterus, and colon by qRT-PCR. We observe heterogeneity in viral load and viral cytopathic effects among various organ systems, between individuals and within the same patient. In a patient with a history of kidney transplant and under immunosuppressant therapy, we observe an unusually high viral load in lung tissue by RNA-ISH, IHC and qRT-PCR. SARS-CoV-2 virus is also detected in this patent's kidney, liver and uterus. We find ACE2, TMPRSS2 and AR expression to overlap with the infection sites. Conclusions: This study portrays the impact of dispersed SARS-CoV-2 infection in diverse organ systems, thereby facilitating avenues for systematic therapeutic approaches.

Diffuse alveolar damage (DAD) resulting from coronavirus disease 2019 Infection is Morphologically Indistinguishable from Other Causes of DAD.

AIMS: Diffuse alveolar damage (DAD) is a ubiquitous finding in inpatient coronavirus disease 2019 (COVID-19)-related deaths, but recent reports have also described additional atypical findings, including vascular changes. An aim of this study was to assess lung autopsy findings in COVID-19 inpatients, and in untreated severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-positive individuals who died in the community, in order to understand the relative impact of medical intervention on lung histology. Additionally, we aimed to investigate whether COVID-19 represents a unique histological variant of DAD by comparing the pathological findings with those of uninfected control patients. METHODS AND RESULTS: Lung sections from autopsy cases were reviewed by three pulmonary pathologists, including two who were blinded to patient cohort. The cohorts included four COVID-19 inpatients, four cases with postmortem SARS-CoV-2 diagnoses who died in the community, and eight SARS-CoV-2-negative control cases. DAD was present in all but one SARS-CoV-2-positive patient, who was asymptomatic and died in the community. Although SARS-CoV-2-positive patients were noted to have more focal perivascular inflammation/endothelialitis than control patients, there were no significant differences in the presence of hyaline membranes, fibrin thrombi, airspace organisation, and 'acute fibrinous and organising pneumonia'-like intra-alveolar fibrin deposition between the cohorts. Fibrinoid vessel wall necrosis, haemorrhage and capillaritis were not features of COVID-19-related DAD. CONCLUSIONS: DAD is the primary histological manifestation of severe lung disease in COVID-19 patients who die both in hospital and in the community, suggesting no contribution of hyperoxaemic mechanical ventilation to the histological changes. There are no distinctive morphological features with which to confidently differentiate COVID-19-related DAD from DAD due to other causes.

Ultrastructural Evidence for Direct Renal Infection with SARS-CoV-2.

BACKGROUND: A significant fraction of patients with coronavirus disease 2019 (COVID-19) display abnormalities in renal function. Retrospective studies of patients hospitalized with COVID-19 in Wuhan, China, report an incidence of 3%-7% progressing to ARF, a marker of poor prognosis. The cause of the renal failure in COVID-19 is unknown, but one hypothesized mechanism is direct renal infection by the causative virus, SARS-CoV-2. METHODS: We performed an autopsy on a single patient who died of COVID-19 after open repair of an aortic dissection, complicated by hypoxic respiratory failure and oliguric renal failure. We used light and electron microscopy to examine renal tissue for evidence of SARS-CoV-2 within renal cells. RESULTS: Light microscopy of proximal tubules showed geographic isometric vacuolization, corresponding to a focus of tubules with abundant intracellular viral arrays. Individual viruses averaged 76 µm in diameter and had an envelope studded with crown-like, electron-dense spikes. Vacuoles contained double-membrane vesicles suggestive of partially assembled virus. CONCLUSIONS: The presence of viral particles in the renal tubular epithelium that were morphologically identical to SARS-CoV-2, and with viral arrays and other features of virus assembly, provide evidence of a productive direct infection of the kidney by SARS-CoV-2. This finding offers confirmatory evidence that direct renal infection occurs in the setting of AKI in COVID-19. However, the frequency and clinical significance of direct infection in COVID-19 is unclear. Tubular isometric vacuolization observed with light microscopy, which correlates with double-membrane vesicles containing vacuoles observed with electronic microscopy, may be a useful histologic marker for active SARS-CoV-2 infection in kidney biopsy or autopsy specimens.

Polypoidal giant cancer cells in metastatic castration-resistant prostate cancer: observations from the Michigan Legacy Tissue Program.

Despite early diagnosis and established protocols, a subset of prostate cancer patients will eventually be categorized as castration-resistant prostate cancer. Recently, it has been reported that these multi-modal therapy cases may harbor a special subset of cancer cells termed as polypoidal giant cancer cells (PGCC). These cells are phenotypically described either as possessing highly irregular polylobated nuclei or multiple pleomorphic nuclei. To identify and characterize the distribution of these cells, we created a cohort of 5 randomly selected cases of multi-modal therapy failure prostate cancer (16 selected non-osseous and osseous tumor sites) enrolled in Michigan Legacy Tissue Program. In all cases, specific "regions of interest" or "hot spots" within tumor areas showing an increased proportion of these multi-nucleated/polylobated cells under light microscopy were labeled as PGCC-rich area. On microscopic evaluation, overall mean count of PGCC was 42.4 ± 3.91 with case 2 in the study cohort with the highest number of average PGCC count of 17 ± 4.04. Site wise analysis showed retroperitoneal lymph node as the tissue with highest number of average PGCC number/site (5.0 ± 0.32). On correlating the average number of PGCC recorded with the time elapsed from last dose of chemotherapy administered to autopsy, the spearman correlation value (R) was 0.67, but the result was not statistically significant (p = 0.22). A systematic assessment of PGCC in a large stratified cohort of prostate cancer patients integrated with various histopathological and clinical parameters along with discovery of specific biomarkers for PGCC are the future studies suggested.

Sudden Death Due to Calcifying Pseudoneoplasm of the Neuraxis: A Case Report and a Review of Sudden Death Due to Undiagnosed Central Nervous System Mass Lesions.

We present a case of a 22-year-old man who died unexpectedly after a seizure due to a previously undiagnosed calcifying pseudoneoplasm of the neuraxis (CAPNON). Calcifying pseudoneoplasm of the neuraxis is a rare entity, and this is, to our knowledge, the first described case of sudden death due to CAPNON. Sudden death due to undiagnosed central nervous system mass lesions is rare, and most cases are attributable to hemorrhage, hydrocephalus, or increased intracranial pressure due to mass effect. Seizure is a rare cause of sudden death due to central nervous system mass lesions. This case highlights that mass lesions may cause sudden death due to seizure, even without other pathologic evidence of a cause of death, such as hemorrhage or edema. Furthermore, benign, reactive, and low-grade mass lesions may cause sudden death due to seizure. Seizure should remain in the autopsy differential as a cause of death, even where there is no pathologically evident mechanism by which a mass lesion caused death.

Plasmacytoid urothelial carcinoma: a rapid autopsy case report with unique clinicopathologic and genomic profile.

BACKGROUND: Rapid ("warm") autopsies of patients with advanced metastatic cancer provide important insight into the natural history, pathobiology and histomorphology of disease in treatment-resistant tumors. Plasmacytoid urothelial carcinoma (PUC) is a rare variant of urothelial carcinoma characterized by neoplastic cells morphologically resembling plasma cells. PUC is typically aggressive, high-stage at presentation, and associated with poor outcomes. Recurrence is common in PUC, with the majority of recurrences occurring in the peritoneum. CASE PRESENTATION: Here, we report rapid autopsy findings from a patient with recurrent PUC. The patient had persistent pain after cystoprostatectomy, although initial post-operative imaging showed no evidence of disease. Imaging obtained shortly before his death showed only subtle growth along vascular tissue planes; however, extensive disease was seen on autopsy. Plasmacytoid tumor cells formed sheets involving many serosal surfaces. Molecular interrogation confirmed a mutation in CDH1 exon 12 leading to early truncation of the CDH1 protein in the tumor cells. CONCLUSIONS: The sheet-like growth pattern of PUC makes early phases of disease spread much more difficult to capture on cross-sectional imaging. Alternative forms of surveillance may be required for detection of recurrent PUC, and providers may need to treat based on symptoms and clinical suspicion.

A Retrospective Study of the Investigation of Homicidal Childhood Asphyxial Deaths.

As one of the leading causes of traumatic deaths in newborns, infants, and young children, there is no anatomic or microscopic feature that is pathognomonic for asphyxial deaths. Instead, pathologists rely on investigation information, including confessions and/or witness statements, and potential evidence at the scene. Twenty cases of homicidal newborn, infant, and young children asphyxial deaths were reviewed, which included death and police investigation reports and autopsy reports, as well as histology slides of lung sections. This series of homicidal asphyxial deaths highlight that, in a vast majority of such cases, the final cause and manner of death rulings are dependent on confession by the perpetrator. Furthermore, this series highlights the possible role of histology to help forensic pathologists better certify asphyxial deaths. Finally, this series emphasizes important investigation points and considerations at autopsy during the investigation of asphyxial deaths in newborns, infants, and young children.

Micro Disasters: The Case of Serial Killer Jeffrey Dahmer.

Disasters are commonly experienced as major devastating events that exceed the resources of an agency to respond, with effects emanating throughout a community or region. There are, however, those events that are more measured, more subtle, and with few actual deaths, which still distract investigators from their daily duties and routines and project long lasting and crippling effects to a community or nation. Disasters can occur from natural forces or be the result of human activity. Most forensic pathologists who practice over a significant time will encounter one or the other types of disaster, sometimes more than a few. In my own career, I have witnessed large-scale disasters, such as hundreds of deaths occurring as the result of a major heat wave, to small-scale disasters such as factory explosions or small airplane crashes at sea-each with their own challenges. In addition to the extent of the initial disaster, many require the detailed, exhaustive evidentiary recovery and examination of a crime scene. The Jeffrey Dahmer case, although only involving 11 actual victims, required a major disaster response, and continues to influence and affect a community over 25 years later.

Prostatic Adenocarcinoma With Hormone Exposure Related Changes in a Patient With Hepatic Cirrhosis - Value of Autopsy in a Case Report.

Hepatic cirrhosis is commonly associated with hyperestrogenism. Previous studies have reported morphologic changes in benign and malignant prostate tissue exposed to estrogen or anti-androgens. To our knowledge, histopathologic features of prostatic adenocarcinoma in patients with cirrhosis have not been well-reported. We present a case of incidental, but pathologically significant, prostatic adenocarcinoma detected on autopsy in a 67-year-old male patient with cirrhosis and spider angiomata. The morphologic and immunohistochemical features (including variable ERG expression) of the prostatic adenocarcinoma were consistent with hormone exposure related changes, suggesting that cirrhosis-induced elevated estrogen-to-testosterone ratio and exogenous hormone therapy might induce similar phenotypes.

Comprehensive Immunophenotypic Characterization of Adult and Fetal Testes, the Excretory Duct System, and Testicular and Epididymal Appendages.

The immunophenotype of a normal testis and the excretory duct system has not been studied comprehensively in fetal and adult patients without testicular disease or hormonal manipulation so far. In addition, testicular (TA) and epididymal (EA) appendages are frequent paratesticular structures without previously reported comprehensive immunophenotypic studies. Immunohistochemistry for multiple markers, including the androgen receptor (AR), the estrogen receptor (ER), the progesterone receptor (PR), the prostate-specific antigen, the prostate-specific membrane antigen, PAX8, WT1, calretinin, CK7, CK20, OCT4, SALL4, and CD117, was performed on full sections of testicular/paratesticular tissue from a large cohort of adult and fetal autopsy patients. In contrast to adult germ cells (GC), fetal GC strongly express OCT4 and CD117, although the expression of these proteins is lost in the early postnatal period; SALL4, in contrast, is expressed in both fetal and adult GC, with only weak and focal expression in adult patients. Fetal Sertoli cells (SC) express WT1 and calretinin strongly and diffusely, in contrast to adult SC. Both fetal and adult excretory duct systems express CK7 and PAX8 with frequent AR coexpression, and all 3 main segments of the excretory duct system (ductuli efferentes, epididymis, and vas deferens) have unique immunophenotypes. The rete testis also has a unique immunohistochemical expression pattern, which includes strong expression of CK7, PAX8, WT1, calretinin, and AR. Finally, of the adult autopsy patients examined, 80% had a TA, and 60% had an EA; these paratesticular structures occurred at stereotypical locations, demonstrated reproducible morphologic features, and had a unique immunophenotype relative to other studied structures, with strong CK7, PAX8, WT1, AR, ER, and PR coexpression. The testis and the paratestis may be involved by diverse neoplastic and non-neoplastic processes, and knowledge of the immunophenotypic expression spectrum of these tissues may aid in clinical diagnosis and advance our understanding of the pathogenesis of both oncologic and nononcologic disease processes.

Whole-Exome Sequencing Reveals Mutations in Genes Linked to Hemophagocytic Lymphohistiocytosis and Macrophage Activation Syndrome in Fatal Cases of H1N1 Influenza.

BACKGROUND: Severe H1N1 influenza can be lethal in otherwise healthy individuals and can have features of reactive hemophagocytic lymphohistiocytosis (HLH). HLH is associated with mutations in lymphocyte cytolytic pathway genes, which have not been previously explored in H1N1 influenza. METHODS: Sixteen cases of fatal influenza A(H1N1) infection, 81% with histopathologic hemophagocytosis, were identified and analyzed for clinical and laboratory features of HLH, using modified HLH-2004 and macrophage activation syndrome (MAS) criteria. Fourteen specimens were subject to whole-exome sequencing. Sequence alignment and variant filtering detected HLH gene mutations and potential disease-causing variants. Cytolytic function of the PRF1 p.A91V mutation was tested in lentiviral-transduced NK-92 natural killer (NK) cells. RESULTS: Despite several lacking variables, cases of influenza A(H1N1) infection met 44% and 81% of modified HLH-2004 and MAS criteria, respectively. Five subjects (36%) carried one of 3 heterozygous LYST mutations, 2 of whom also possessed the p.A91V PRF1 mutation, which was shown to decrease NK cell cytolytic function. Several patients also carried rare variants in other genes previously observed in MAS. CONCLUSIONS: This cohort of fatal influenza A(H1N1) infections confirms the presence of hemophagocytosis and HLH pathology. Moreover, the high percentage of HLH gene mutations suggests they are risk factors for mortality among individuals with influenza A(H1N1) infection.

Anaplastic lymphoma kinase-positive anaplastic large cell lymphoma presenting as retroperitoneal fibrosis.

Retroperitoneal fibrosis is a rare fibroinflammatory condition involving the abdominal aorta, iliac vessels, and ureters that carries an association with several other autoimmune conditions. Most cases of retroperitoneal fibrosis are thought to be idiopathic. The disorder can affect all age groups but is most common in persons between the ages of 50 and 70 years. A subset of cases is associated with an underlying immunohematologic abnormality including lymphoma. We describe in this case report a highly unusual presentation of a young woman who died with a diagnosis of "idiopathic retroperitoneal fibrosis" based on multiple biopsy procedures. Postmortem examination, however, revealed disseminated anaplastic lymphoma kinase-positive anaplastic large cell lymphoma. The clinical and histopathologic importance of this very unusual presentation of anaplastic lymphoma kinase-positive anaplastic large cell lymphoma with retroperitoneal fibrosis is discussed.

Neck and scleral hemorrhage in drowning.

The determination of the cause and manner of death for a body recovered from the water can be difficult because of a lack of autopsy findings specific for drowning. This case report describes a 30-year-old man found submerged at the bottom of a hotel pool. An autopsy revealed scleral hemorrhages and fascial hemorrhages of multiple muscles of the anterior and posterior neck bilaterally. No evidence of traumatic injury was on the surface of the body. An investigation by law enforcement found no evidence of foul play. The occurrence of petechial and neck hemorrhage in a body recovered from the water is controversial, and a review of this literature will be given. We suggest that fascial hemorrhages of the muscles of the neck, as well as cephalic hemorrhages, can be explained by drowning-related elevated central venous pressure that is communicated to the head through the valveless veins of the neck.

Autopsy findings in eight patients with fatal H1N1 influenza.

A novel H1N1 influenza A virus emerged in April 2009, and rapidly reached pandemic proportions. We report a retrospective observational case study of pathologic findings in 8 patients with fatal novel H1N1 infection at the University of Michigan Health Systems (Ann Arbor) compared with 8 age-, sex-, body mass index-, and treatment-matched control subjects. Diffuse alveolar damage (DAD) in acute and organizing phases affected all patients with influenza and was accompanied by acute bronchopneumonia in 6 patients. Organizing DAD with established fibrosis was present in 1 patient with preexisting granulomatous lung disease. Only 50% of control subjects had DAD. Peripheral pulmonary vascular thrombosis occurred in 5 of 8 patients with influenza and 3 of 8 control subjects. Cytophagocytosis was seen in all influenza-related cases. The autopsy findings in our patients with novel H1N1 influenza resemble other influenza virus infections with the exception of prominent thrombosis and hemophagocytosis. The possibility of hemophagocytic syndrome should be investigated in severely ill patients with H1N1 infection.

Infant deaths associated with human parechovirus infection in Wisconsin.

BACKGROUND: From December 1987 through August 2004, lung tissue, nasopharyngeal swabs, and colon swab specimens obtained during 1263 autopsies of infants and young children were examined to assess the role of viruses in deaths of children aged <2 years. METHODS: Multiple cell cultures were used to isolate viruses. With 4 exceptions, virus isolates were identified by neutralization, immunofluorescence assay, or enzyme immunoassay. RNA extracted from these 4 isolates and associated autopsy specimens was tested using parechovirus-specific real-time polymerase chain reaction (RT-PCR) and sequencing assays. RESULTS: Specimens from 426 (34%) autopsies were positive for at least 1 virus; enteroviruses and adenoviruses were the most commonly identified. Human parechoviruses (HPeVs) were identified antigenically in isolates from 18 decedents (all HPeV type 1) and by RT-PCR in isolates and multiple autopsy specimens from 4 decedents with untypeable virus isolates. Sequencing of the VP1 region identified these 4 HPeVs as HPeV type 3 (n = 3) and HPeV type 6 (n = 1). Despite the detection of HPeV, the deaths of decedents 3 and 4 were determined to have been from noninfectious causes. CONCLUSIONS: These are the first confirmed HPeV type 3 and HPeV type 6 detections in the United States. This is also the initial report of fatal cases with associated HPeV type 3 infection. These results support prior findings associating HPeVs with serious disease in young children. Clinical testing for HPeVs and routine HPeV surveillance by public health laboratories will help determine the burden of disease caused by HPeVs.

Retinal hemorrhages in children following fatal motor vehicle crashes: a case series.

OBJECTIVE: To demonstrate the severity of ocular findings in young children who died of injuries due to motor vehicle crashes. METHODS: Case series of 10 children younger than 3 years who were fatally injured in motor vehicle crashes between January 1, 1994, and December 31, 2002. All children underwent autopsy that included eye examination. All available medical and autopsy records, pathology slides and photographs, and police and traffic department reports were reviewed for each case. RESULTS: Eight patients had retinal hemorrhages, which extended into the periphery in 13 eyes and were bilateral in 7 patients. Three patients had elevated circular retinal folds. Six patients had hemorrhages below the internal limiting membrane, but no patients had deeper splitting of the retina. Nine patients had optic nerve sheath hemorrhages. CONCLUSION: The association of extensive, sometimes severe, ocular hemorrhages with fatal accidental trauma, compared with previous reports of accidental trauma with no or few hemorrhages, indicates the severity of injury required to cause hemorrhages of this magnitude.

Selecting forensic pathology as a career: a survey of the past with an eye on the future.

BACKGROUND: Each year there are about 30 to 40 physicians who train and become board-certified in the specialty area of forensic pathology, compared with hundreds or thousands in other disciplines. There are not enough board-certified forensic pathologists to cover national need. The National Association of Medical Examiners' (NAME) Forensic Pathology Training Committee conducted a survey of its members to determine which factors influenced them to select forensic pathology as a career, and to offer suggestions about possible recruitment methods in the future. METHODS: Two of the authors developed a 13-question survey form that included questions designed to determine the demographics of the responders, education level at which interest emerged, influential factors in the selection of forensic pathology, exposure to the subject matter of forensic pathology in medical school and residency, opinions about the best educational level for recruitment targeting, and faculty reactions to selection of forensic pathology as a career choice. Comments and suggestions were also solicited. The survey was sent by email to the 552 physician NAME members who have email addresses on file at the NAME Home Office. RESULTS: One hundred sixty-one surveys were returned for a response rate of 29%. Most responders were full-time, board-certified forensic pathologists who had been practicing for an average of 18 years. The most influential factors in developing interest were exposure to forensic pathology in residency training and the influence of a professor or mentor. Medical school was the favored education level to target recruitment. Less than half had a forensic pathologist as an autopsy instructor in anatomic pathology residency. The number of responders who were encouraged by faculty to pursue forensic pathology was about the same as the number who were either discouraged or who perceived no particular positive or negative reinforcement. The typical scenario for forensic pathology exposure during anatomic pathology residency was a 4-week rotation at an off-site location from the medical school or hospital, with a mentor that had an adjunct, assistant, associate, or clinical faculty appointment. CONCLUSIONS: If the past predicts the future, it will be important to ensure that pathology residents have a planned and positive exposure to forensic pathology and that forensic pathologist mentors are available to training programs. There are a variety of other methods that might be used for recruitment which include more emphasis on medical students, a more academic approach, and affiliation, emphasizing the scientific nature of the work, integrating forensic pathology more into the ongoing medical school curriculum, improving the anatomic pathology residency autopsy experience, and avoiding possible turnoffs that can be caused by presentation of sensational or unpleasant cases that are not representative of routine daily work. Improved remuneration and building esteem by peers were also cited as critical factors, as was recruitment of more physicians into pathology in general. The Committee intends to develop a plan for recruitment and retention in the field of forensic pathology. Based on the survey data, this will require a conjoined effort with the American Association of Medical Colleges, the Accreditation Council on Graduate Medical Education, the Association of Pathology Chairman, and other entities to enable a planned and multifaceted approach to recruitment and retention in the field.