Temporal changes in pathology and viral RNA distribution in guinea pigs following separate infection with two New World Arenaviruses.

Numerous arenaviruses have been identified throughout the Americas and a subset of these viruses cause viral hemorrhagic fever in humans. This study compared the pathology and viral RNA distribution in Hartley guinea pigs challenged with two human-disease causing New World arenaviruses, Junin virus (JUNV) or Guanarito virus (GTOV). Histopathologic analysis and RNA in situ hybridization revealed similar pathology and viral RNA distribution for both groups of animals challenged with either JUNV or GTOV on days 3, 7, 10 and 12 post exposure (PE). Gross lesions were first observed on day 7 and primarily involved the lungs and liver. The most severe histologic lesions occurred in the lymph nodes, spleen, and thymus and included lymphoid depletion and necrosis which increased in severity over time. Extensive necrosis was also observed in the bone marrow on day 12. Minimal to mild inflammation with and without necrosis was observed in the choroid plexus of the brain, choroid of the eye, intestinal tract, lung and adrenal gland. Significant liver lesions were rare, consisting predominantly of hepatocyte vacuolation. Viral RNA labeling was identified in nearly all organs examined, was often extensive in certain organs and generally increased over time starting on day 7. Our data demonstrate the guinea pig may serve as a useful model to study New World arenavirus infection in humans and for the evaluation and development of medical countermeasures.

Eastern equine encephalitis virus rapidly infects and disseminates in the brain and spinal cord of cynomolgus macaques following aerosol challenge.

Eastern equine encephalitis virus (EEEV) is mosquito-borne virus that produces fatal encephalitis in humans. We recently conducted a first of its kind study to investigate EEEV clinical disease course following aerosol challenge in a cynomolgus macaque model utilizing the state-of-the-art telemetry to measure critical physiological parameters. Here, we report the results of a comprehensive pathology study of NHP tissues collected at euthanasia to gain insights into EEEV pathogenesis. Viral RNA and proteins as well as microscopic lesions were absent in the visceral organs. In contrast, viral RNA and proteins were readily detected throughout the brain including autonomic nervous system (ANS) control centers and spinal cord. However, despite presence of viral RNA and proteins, majority of the brain and spinal cord tissues exhibited minimal or no microscopic lesions. The virus tropism was restricted primarily to neurons, and virus particles (~61-68 nm) were present within axons of neurons and throughout the extracellular spaces. However, active virus replication was absent or minimal in majority of the brain and was limited to regions proximal to the olfactory tract. These data suggest that EEEV initially replicates in/near the olfactory bulb following aerosol challenge and is rapidly transported to distal regions of the brain by exploiting the neuronal axonal transport system to facilitate neuron-to-neuron spread. Once within the brain, the virus gains access to the ANS control centers likely leading to disruption and/or dysregulation of critical physiological parameters to produce severe disease. Moreover, the absence of microscopic lesions strongly suggests that the underlying mechanism of EEEV pathogenesis is due to neuronal dysfunction rather than neuronal death. This study is the first comprehensive investigation into EEEV pathology in a NHP model and will provide significant insights into the evaluation of countermeasure.

Ebola virus persistence and disease recrudescence in the brains of antibody-treated nonhuman primate survivors.

Effective therapeutics have been developed against acute Ebola virus disease (EVD) in both humans and experimentally infected nonhuman primates. However, the risk of viral persistence and associated disease recrudescence in survivors receiving these therapeutics remains unclear. In contrast to rhesus macaques that survived Ebola virus (EBOV) exposure in the absence of treatment, we discovered that EBOV, despite being cleared from all other organs, persisted in the brain ventricular system of rhesus macaque survivors that had received monoclonal antibody (mAb) treatment. In mAb-treated macaque survivors, EBOV persisted in macrophages infiltrating the brain ventricular system, including the choroid plexuses. This macrophage infiltration was accompanied by severe tissue damage, including ventriculitis, choroid plexitis, and meningoencephalitis. Specifically, choroid plexus endothelium-derived EBOV infection led to viral persistence in the macaque brain ventricular system. This resulted in apoptosis of ependymal cells, which constitute the blood-cerebrospinal fluid barrier of the choroid plexuses. Fatal brain-confined recrudescence of EBOV infection manifested as severe inflammation, local pathology, and widespread infection of the ventricular system and adjacent neuropil in some of the mAb-treated macaque survivors. This study highlights organ-specific EBOV persistence and fatal recrudescent disease in rhesus macaque survivors after therapeutic treatment and has implications for the long-term follow-up of human survivors of EVD.

Hamsters Expressing Human Angiotensin-Converting Enzyme 2 Develop Severe Disease following Exposure to SARS-CoV-2.

The rapid emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has created a global health emergency. While most human disease is mild to moderate, some infections lead to a severe disease characterized by acute respiratory distress, hypoxia, anosmia, ageusia, and, in some instances, neurological involvement. Small-animal models reproducing severe disease, including neurological sequela, are needed to characterize the pathophysiological mechanism(s) of disease and to identify medical countermeasures. Transgenic mice expressing the human angiotensin-converting enzyme 2 (hACE2) viral receptor under the control of the K18 promoter develop severe and lethal respiratory disease subsequent to SARS-CoV-2 intranasal challenge when high viral doses are used. Here, we report on SARS-CoV-2 infection of hamsters engineered to express the hACE2 receptor under the control of the K18 promoter. K18-hACE2 hamsters infected with a relatively low dose of 100 or 1,000 PFU of SARS-CoV-2 developed a severe and lethal disease, with most animals succumbing by day 5 postinfection. Hamsters developed severe lesions and inflammation within the upper and lower respiratory system, including infection of the nasal cavities causing marked destruction of the olfactory epithelium as well as severe bronchopneumonia that extended deep into the alveoli. Additionally, SARS-CoV-2 infection spread to the central nervous system (CNS), including the brain stem and spinal cord. Wild-type (WT) hamsters naturally support SARS-CoV-2 infection, with the primary lesions present in the respiratory tract and nasal cavity. Overall, infection in the K18-hACE2 hamsters is more extensive than that in WT hamsters, with more CNS involvement and a lethal outcome. These findings demonstrate the K18-hACE2 hamster model will be valuable for studying SARS-CoV-2. IMPORTANCE The rapid emergence of SARS-CoV-2 has created a global health emergency. While most human SARS-CoV-2 disease is mild, some people develop severe, life-threatening disease. Small-animal models mimicking the severe aspects of human disease are needed to more clearly understand the pathophysiological processes driving this progression. Here, we studied SARS-CoV-2 infection in hamsters engineered to express the human angiotensin-converting enzyme 2 viral receptor under the control of the K18 promoter. SARS-CoV-2 produces a severe and lethal infection in transgenic hamsters that mirrors the most severe aspects of COVID-19 in humans, including respiratory and neurological injury. In contrast to other animal systems, hamsters manifest disease with levels of input virus more consistent with natural human infection. This system will be useful for the study of SARS-CoV-2 disease and the development of drugs targeting this virus.

Creating locally-resolved mobile-source emissions inputs for air quality modeling in support of an exposure study in Detroit, Michigan, USA.

This work describes a methodology for modeling the impact of traffic-generated air pollutants in an urban area. This methodology presented here utilizes road network geometry, traffic volume, temporal allocation factors, fleet mixes, and emission factors to provide critical modeling inputs. These inputs, assembled from a variety of sources, are combined with meteorological inputs to generate link-based emissions for use in dispersion modeling to estimate pollutant concentration levels due to traffic. A case study implementing this methodology for a large health study is presented, including a sensitivity analysis of the modeling results reinforcing the importance of model inputs and identify those having greater relative impact, such as fleet mix. In addition, an example use of local measurements of fleet activity to supplement model inputs is described, and its impacts to the model outputs are discussed. We conclude that with detailed model inputs supported by local traffic measurements and meteorology, it is possible to capture the spatial and temporal patterns needed to accurately estimate exposure from traffic-related pollutants.

Associations between ozone and morbidity using the Spatial Synoptic Classification system.

BACKGROUND: Synoptic circulation patterns (large-scale tropospheric motion systems) affect air pollution and, potentially, air-pollution-morbidity associations. We evaluated the effect of synoptic circulation patterns (air masses) on the association between ozone and hospital admissions for asthma and myocardial infarction (MI) among adults in North Carolina. METHODS: Daily surface meteorology data (including precipitation, wind speed, and dew point) for five selected cities in North Carolina were obtained from the U.S. EPA Air Quality System (AQS), which were in turn based on data from the National Climatic Data Center of the National Oceanic and Atmospheric Administration. We used the Spatial Synoptic Classification system to classify each day of the 9-year period from 1996 through 2004 into one of seven different air mass types: dry polar, dry moderate, dry tropical, moist polar, moist moderate, moist tropical, or transitional. Daily 24-hour maximum 1-hour ambient concentrations of ozone were obtained from the AQS. Asthma and MI hospital admissions data for the 9-year period were obtained from the North Carolina Department of Health and Human Services. Generalized linear models were used to assess the association of the hospitalizations with ozone concentrations and specific air mass types, using pollutant lags of 0 to 5 days. We examined the effect across cities on days with the same air mass type. In all models we adjusted for dew point and day-of-the-week effects related to hospital admissions. RESULTS: Ozone was associated with asthma under dry tropical (1- to 5-day lags), transitional (3- and 4-day lags), and extreme moist tropical (0-day lag) air masses. Ozone was associated with MI only under the extreme moist tropical (5-day lag) air masses. CONCLUSIONS: Elevated ozone levels are associated with dry tropical, dry moderate, and moist tropical air masses, with the highest ozone levels being associated with the dry tropical air mass. Certain synoptic circulation patterns/air masses in conjunction with ambient ozone levels were associated with increased asthma and MI hospitalizations.

Giant berry aneurysms in a forensic setting: a series of 14 cases.

This is a series reviewing 14 cases of giant saccular aneurysms diagnosed at the Office of the Chief Medical Examiner of New York City collected over an 11-year period. Data collected on all 14 cases included neuropathological findings, comorbidities, and toxicological findings. Of these 14 cases, 8 were in women, and the ages ranged from 3 to 79 years, with a mean and a median of 50 years. Women were overrepresented in the sixth through eighth decades. Of the 14 cases described, 11 presented with a subarachnoid hemorrhage; 3, no hemorrhage; 2, subdural hemorrhage; 8, intraventricular hemorrhage; 2, intracerebral hemorrhage; and 8, more than 1 hemorrhage type. Location of the aneurysms varied with 6 in the left side of the brain, 6 present in the right side of the brain, and 2 at the midline. We described the clinical, pathological, and toxicological findings associated with these giant aneurysms.

Cost-analysis comparison of outpatient see-and-treat hysteroscopy service with other hysteroscopy service models.

STUDY OBJECTIVE: To conduct a cost analysis of 3 different hysteroscopy service models. DESIGN: Decision-analytic model constructed from the UK National Health Service perspective (Canadian Task Force classification III). SETTING: Tertiary-care hospital. PATIENTS: Women undergoing hysteroscopy (N=1109). INTERVENTIONS: Three hysteroscopy service models: outpatient see-and-treat service; outpatient diagnostic hysteroscopy followed by referral for operative hysteroscopy under general anesthesia (outpatient and referral service); and general anesthesia see-and-treat service. MEASUREMENTS AND MAIN RESULTS: Costs were measured in 2008 UK pounds sterling. Of the 3 treatment arms, total costs were lowest with outpatient see-and-treat service. The lower cost of the outpatient see-and-treat service was observed across a number of patient subgroups (age, menopause status, and indication) and when subjected to sensitivity analyses. CONCLUSIONS: Outpatient see-and-treat hysteroscopy was associated with the lowest treatment costs. This service model may reduce the total cost of care in women referred for hysteroscopy.

Age stereotypes in middle-aged through old-old adults.

The primary goal of the study was to compare adult age groups on aging bias, with measures of knowledge of aging in the physical, psychological, and social domains and life satisfaction. The study sample, consisting of 752 men and women, 40 to 95 years of age, was tested using Neugarten, Havighurst, and Tobin's (1961) Life Satisfaction Index (LSI) and Palmore's Facts on Aging Quiz (1998) modified to extract bias toward older adults and categorized into 3 domains: physical, psychological, and social. Independent variables were 3 levels of LSI from the Neugarten et al. quiz. Aging bias was measured among the age groups, gender, life satisfaction, and knowledge of aging domains. Significant bias differences were found in age group, life satisfaction, and knowledge of aging domain variables. Financial, health, and volunteer status interacted with these effects. Among other significant findings, the data indicate that middle-aged adults 40-59 have the most negative bias in the psychological and social domains and the least negative bias in the physical domain compared to the older participants. Both middle aged and old-old adults have the most negative aging biases. These differential aging stereotypes (positive and negative) among the physical, psychological, and social perceptions of aging over adult age groups are interpreted within aging stereotyping and aging self-stereotyping.

The CLEF corpus: semantic annotation of clinical text.

The Clinical E-Science Framework (CLEF) project is building a framework for the capture, integration and presentation of clinical information: for clinical research, evidence-based health care and genotype-meets-phenotype informatics. A significant portion of the information required by such a framework originates as text, even in EHR-savvy organizations. CLEF uses Information Extraction (IE) to make this unstructured information available. An important part of IE is the identification of semantic entities and relationships. Typical approaches require human annotated documents to provide both evaluation standards and material for system development. CLEF has a corpus of clinical narratives, histopathology reports and imaging reports from 20 thousand patients. We describe the selection of a subset of this corpus for manual annotation of clinical entities and relationships. We describe an annotation methodology and report encouraging initial results of inter-annotator agreement. Comparisons are made between different text sub-genres, and between annotators with different skills.

The retina in forensic medicine: applications of ophthalmic endoscopy: the first 100 cases.

The retina reflects a variety of diseases in the living patient. However, the retina is not routinely examined in deceased persons, and therefore it is unknown if routine retinal examination would be a useful adjunct to the forensic autopsy. To examine this issue, the retinae of routine medical examiner cases were examined utilizing an ophthalmic endoscope. The results of the first 100 examinations are reported. Specific attention was given to changes that reflected the postmortem interval, the development of petechiae as related to cardiopulmonary resuscitation, and the association of retinal hemorrhages to subconjunctival hemorrhages. The procedure was helpful in cases of suspected shaken baby syndrome, exsanguination, and carbon monoxide poisoning and in cases with sudden increased intracranial pressure (Terson syndrome). It appears that lividity patterns exist in the retina, and this may be potentially useful in determining body position after death. Some natural disease processes, such as hypertension, were also identified. Finally, the utility of the ophthalmic endoscope as a means of circumventing the problem of corneal clouding is discussed, and ideas for further research using this technology are presented.

Knowledge of aging and life satisfaction among older adults.

Four hundred young-, middle-, and old-old adults responded to a battery of quizzes dealing with life satisfaction and objective aging knowledge in the physical, psychological, and social domains. Analyses incorporated domains of aging knowledge, life satisfaction, age, gender, and demographic variables. Both means difference and regression analyses were computed. Significant age group, gender, and life satisfaction differences were found for the three aging knowledge domains. For successive age groups, knowledge of aging decreased, with females knowing less than males. The greater knowledge of aging, the higher the life satisfaction. The demographic variables education, financial status, health, living arrangement, and volunteerism were significant covariates for knowledge of aging. Results from this study indicate that knowledge of aging in specific domains varies among older adult age groups and is associated with life satisfaction.

Souvenir knife: a retained transcranial knife blade.

Upon necroscopic examination of a homeless male found comatose in the street and pronounced dead at a medical center 12 hours later, a sharp tip of a knife lodged in the right parietal region of his skull was incidentally discovered. The blade transected the diploe and penetrated the cerebral cortex. Subsequent police investigation revealed that this was the remnant of a stabbing attempt on his life several months prior to his death. The cause of death was determined to be unrelated to the metallic blade fragment, thus making it a truly incidental and rare finding of a "souvenir knife." Nevertheless, since the injury sustained in the stabbing was potentially life threatening, the investigation into that assault was reopened.A case report is presented, along with a brief review of the literature on "souvenir objects."

Unlocking the brain: attempts to improve mental function of microcephalic retarded children by "craniotomy".

"Is it sufficient glory to don a white apron and swing a carbolized knife and is therein a sufficient indication to let daylight into a deformed cranium and on top of a hopelessly defective brain, and to proclaim success because the victim consented not to die of the assault? Such rash feats of indiscriminate surgery, if continued, moreover in the presence of fourteen deaths in thirty-three cases, are stains on your hands and sins on your souls. No ocean of soap and water will cleanse those hands, no power of corrosive sublimate will disinfect the souls." These passionate words, delivered by Abraham Jacobi, the father of American pediatrics, at the International Congress in Rome in 1893, and later in the article "Non nocere" (42), epitomize the growing antagonism to the attempts by many prominent surgeons to improve the gloomy fate of severely retarded, microcephalic children by "liberating" their brains from their presumed bony chains by "linear craniotomy." This article portrays how the fallacious 19th-century concepts of the relationship between the capacity of the cranial cavity and intelligence, backed by "scientific racism" together with the changing attitude toward retarded and malformed children, generated a surgical solution for microcephalic idiocy. It describes how hopeful surgeons, neurologists, and pediatricians, encouraged by the advances in anesthesia and asepsis, lost their judgment and disregarded logic and evidence. They generated a wave of enthusiasm and hope that soared from the United States and France through the British Isles, Europe, and as far as Ceylon and Australia to end in a ripple of bitter disgrace under caustic criticism, leaving a scar on the recently budding field of neurological surgery.

Mutations of the BRAF gene in human cancer.

Cancers arise owing to the accumulation of mutations in critical genes that alter normal programmes of cell proliferation, differentiation and death. As the first stage of a systematic genome-wide screen for these genes, we have prioritized for analysis signalling pathways in which at least one gene is mutated in human cancer. The RAS RAF MEK ERK MAP kinase pathway mediates cellular responses to growth signals. RAS is mutated to an oncogenic form in about 15% of human cancer. The three RAF genes code for cytoplasmic serine/threonine kinases that are regulated by binding RAS. Here we report BRAF somatic missense mutations in 66% of malignant melanomas and at lower frequency in a wide range of human cancers. All mutations are within the kinase domain, with a single substitution (V599E) accounting for 80%. Mutated BRAF proteins have elevated kinase activity and are transforming in NIH3T3 cells. Furthermore, RAS function is not required for the growth of cancer cell lines with the V599E mutation. As BRAF is a serine/threonine kinase that is commonly activated by somatic point mutation in human cancer, it may provide new therapeutic opportunities in malignant melanoma.