Influenza Virus Aerosols in Human Exhaled Breath: Particle Size, Culturability, and Effect of Surgical Masks

The CDC recommends that healthcare settings provide influenza patients with facemasks as a means of reducing transmission to staff and other patients, and a recent report suggested that surgical masks can capture influenza virus in large droplet spray. However, there is minimal data on influenza virus aerosol shedding, the infectiousness of exhaled aerosols, and none on the impact of facemasks on viral aerosol shedding from patients with seasonal influenza. We collected samples of exhaled particles (one with and one without a facemask) in two size fractions (“coarse”>5 µm, “fine”≤5 µm) from 37 volunteers within 5 days of seasonal influenza onset, measured viral copy number using quantitative RT-PCR, and tested the fine-particle fraction for culturable virus. Fine particles contained 8.8 (95% CI 4.1 to 19) fold more viral copies than did coarse particles. Surgical masks reduced viral copy numbers in the fine fraction by 2.8 fold (95% CI 1.5 to 5.2) and in the coarse fraction by 25 fold (95% CI 3.5 to 180). Overall, masks produced a 3.4 fold (95% CI 1.8 to 6.3) reduction in viral aerosol shedding. Correlations between nasopharyngeal swab and the aerosol fraction copy numbers were weak (r = 0.17, coarse; r = 0.29, fine fraction). Copy numbers in exhaled breath declined rapidly with day after onset of illness. Two subjects with the highest copy numbers gave culture positive fine particle samples. Surgical masks worn by patients reduce aerosols shedding of virus. The abundance of viral copies in fine particle aerosols and evidence for their infectiousness suggests an important role in seasonal influenza transmission. Monitoring exhaled virus aerosols will be important for validation of experimental transmission studies in humans

Influenza Virus in Human Exhaled Breath: An Observational Study

Background: Recent studies suggest that humans exhale fine particles during tidal breathing but little is known of their composition, particularly during infection. Methodology/Principal Findings: We conducted a study of influenza infected patients to characterize influenza virus and particle concentrations in their exhaled breath. Patients presenting with influenza-like-illness, confirmed influenza A or B virus by rapid test, and onset within 3 days were recruited at three clinics in Hong Kong, China. We collected exhaled breath from each subject onto Teflon filters and measured exhaled particle concentrations using an optical particle counter. Filters were analyzed for influenza A and B viruses by quantitative polymerase chain reaction (qPCR). Twelve out of thirteen rapid test positive patients provided exhaled breath filter samples (7 subjects infected with influenza B virus and 5 subjects infected with influenza A virus). We detected influenza virus RNA in the exhaled breath of 4 (33%) subjects–three (60%) of the five patients infected with influenza A virus and one (14%) of the seven infected with influenza B virus. Exhaled influenza virus RNA generation rates ranged from <3.2 to 20 influenza virus RNA particles per minute. Over 87% of particles exhaled were under 1 µm in diameter. Conclusions: These findings regarding influenza virus RNA suggest that influenza virus may be contained in fine particles generated during tidal breathing, and add to the body of literature suggesting that fine particle aerosols may play a role in influenza transmission

Inactivation of Poxviruses by Upper-Room UVC Light in a Simulated Hospital Room Environment

In the event of a smallpox outbreak due to bioterrorism, delays in vaccination programs may lead to significant secondary transmission. In the early phases of such an outbreak, transmission of smallpox will take place especially in locations where infected persons may congregate, such as hospital emergency rooms. Air disinfection using upper-room 254 nm (UVC) light can lower the airborne concentrations of infective viruses in the lower part of the room, and thereby control the spread of airborne infections among room occupants without exposing occupants to a significant amount of UVC. Using vaccinia virus aerosols as a surrogate for smallpox we report on the effectiveness of air disinfection, via upper-room UVC light, under simulated real world conditions including the effects of convection, mechanical mixing, temperature and relative humidity. In decay experiments, upper-room UVC fixtures used with mixing by a conventional ceiling fan produced decreases in airborne virus concentrations that would require additional ventilation of more than 87 air changes per hour. Under steady state conditions the effective air changes per hour associated with upper-room UVC ranged from 18 to 1000. The surprisingly high end of the observed range resulted from the extreme susceptibility of vaccinia virus to UVC at low relative humidity and use of 4 UVC fixtures in a small room with efficient air mixing. Increasing the number of UVC fixtures or mechanical ventilation rates resulted in greater fractional reduction in virus aerosol and UVC effectiveness was higher in winter compared to summer for each scenario tested. These data demonstrate that upper-room UVC has the potential to greatly reduce exposure to susceptible viral aerosols. The greater survival at baseline and greater UVC susceptibility of vaccinia under winter conditions suggest that while risk from an aerosol attack with smallpox would be greatest in winter, protective measures using UVC may also be most efficient at this time. These data may also be relevant to influenza, which also has improved aerosol survival at low RH and somewhat similar sensitivity to UVC

Interobserver agreement in dysplasia grading: toward an enhanced gold standard for clinical pathology trials

Objective: Interobserver agreement in the context of oral epithelial dysplasia (OED) grading has been notoriously unreliable and can impose barriers for developing new molecular markers and diagnostic technologies. This paper aimed to report the details of a 3-stage histopathology review and adjudication process with the goal of achieving a consensus histopathologic diagnosis of each biopsy. Study Design: Two adjacent serial histologic sections of oral lesions from 846 patients were independently scored by 2 different pathologists from a pool of 4. In instances where the original 2 pathologists disagreed, a third, independent adjudicating pathologist conducted a review of both sections. If a majority agreement was not achieved, the third stage involved a face-to-face consensus review. Results: Individual pathologist pair κ values ranged from 0.251 to 0.706 (fair-good) before the 3-stage review process. During the initial review phase, the 2 pathologists agreed on a diagnosis for 69.9% of the cases. After the adjudication review by a third pathologist, an additional 22.8% of cases were given a consensus diagnosis (agreement of 2 out of 3 pathologists). After the face-to-face review, the remaining 7.3% of cases had a consensus diagnosis. Conclusions: The use of the defined protocol resulted in a substantial increase (30%) in diagnostic agreement and has the potential to improve the level of agreement for establishing gold standards for studies based on histopathologic diagnosis

Severe 2010 Cold-Water Event Caused Unprecedented Mortality to Corals of the Florida Reef Tract and Reversed Previous Survivorship Patterns

Background Coral reefs are facing increasing pressure from natural and anthropogenic stressors that have already caused significant worldwide declines. In January 2010, coral reefs of Florida, United States, were impacted by an extreme cold-water anomaly that exposed corals to temperatures well below their reported thresholds (16°C), causing rapid coral mortality unprecedented in spatial extent and severity. Methodology/Principal Findings Reef surveys were conducted from Martin County to the Lower Florida Keys within weeks of the anomaly. The impacts recorded were catastrophic and exceeded those of any previous disturbances in the region. Coral mortality patterns were directly correlated to in-situ and satellite-derived cold-temperature metrics. These impacts rival, in spatial extent and intensity, the impacts of the well-publicized warm-water bleaching events around the globe. The mean percent coral mortality recorded for all species and subregions was 11.5% in the 2010 winter, compared to 0.5% recorded in the previous five summers, including years like 2005 where warm-water bleaching was prevalent. Highest mean mortality (15%–39%) was documented for inshore habitats where temperatures were \u3c11°C for prolonged periods. Increases in mortality from previous years were significant for 21 of 25 coral species, and were 1–2 orders of magnitude higher for most species. Conclusions/Significance The cold-water anomaly of January 2010 caused the worst coral mortality on record for the Florida Reef Tract, highlighting the potential catastrophic impacts that unusual but extreme climatic events can have on the persistence of coral reefs. Moreover, habitats and species most severely affected were those found in high-coral cover, inshore, shallow reef habitats previously considered the “oases” of the region, having escaped declining patterns observed for more offshore habitats. Thus, the 2010 cold-water anomaly not only caused widespread coral mortality but also reversed prior resistance and resilience patterns that will take decades to recover

EIN ALTKROATISCHES GRABERFELD AUF DEM BERGE SPAS BEI KNIN

Autor objavljuje rezultate istraživanja starohrvatske nekropole na brdu Spasu kod Knina (otkriveno ukupno 228 grobova). Na temelju prikupljenih podataka tijekom višegodišnjih arheoloških istraživanja i sprovedene analize grobne arhitekture, pogrebnih običaja i grobnih nalaza utvrđeno je vrijeme ukopavanja na toj nekropoli i uputilo na mogući prostorni smještaj naselja kojemu je to groblje pripadalo. Završna razmatranja objelodanjenih grobnih nalaza otkrivaju niz novih spoznaja o gospoda.stvenim mogućnostima stanovništva, njihov kulturni i duhovni život u 9, 10. i 11. stoljeću. Osim starohrvatskih grobova na Spasu su otkopani i temeljni ostaci arhitekture, za koje se utvrdilo da pripadaju rimskom naselju, vjerojatno ostacima stare Ninie, odnosno Curcuma. Sve to popraćeno je obilnom dokumentacijskom građom i katalogom grobova.Westlich der Stadt Knin erstreckt sich der Berg Spas. Auf seinem siidlichen Teil erhebt sich die mittelalterliche Festung Knin, und an den nord lichen Teil schlieBt sich das Hochplateau des Berges Spas an, das im horizontalen Querschnitt cinE\u27 birnentormige Form hat. Spas ist anfangs des 18. lahrhunderts durch einen kiinstlichen Durchstich von der Festung getrennt worden. Im nordlichen, breitesten Teil des Plateaus wurde ein altkroatisches Graberfeld entdeckt, mit insgesamt 228 Grabern. Das erforsci1te Gebiet umfaBt etwa 1200 m2 Oberflache. Es wurden neben Grabern auch Fundamente von Bauwerken aus vorktoatischer Zeit gefunden, was bezeugt, daB es sich hier um eine komplexe archaologische Fundstatte handelt. Im ersten Teil seiner Abhandlung beschreibt. der Verfasser die Lage des Fundortes, seine strategische Bedeutung, und den geschichtlichen Ablauf der Forschungen an diesem Ort. AuBerdem erwahnt er unter anderen bedeutenden Funden auch die Entdeckung von Fundamenten von Bauwerken , fUr die er aufgrund von Fragmenten steinernen Skulpturen, Ziegeln und Fragmenten von KeramikgefaBen festelIen konnte, daB sie einer romisehen Ansiedlung angehorten. Durch Suchgrabung wurde festgestelIt, daB die Reste dieser Bauwerke sich auch auf den iibrigen Teil des Plateaus erstrecken, der bis jetzt nicht erforscht worden ist. Diese Entdeckungen, zusammen mit einem groBen, schon friiher untersuchten Graberfeld am ostlichen FuBe dieses Berges, genannt »Greblje«, sind ein starkes Argumentfi.ir die Annahme jener Forscher, die der Meinung sind, daB die alte, antike Stadtsiedlung Ninia, die von der Schriftstellern der Antike erwahnt wird , sich an der SteIle der heutigen Stadt Knin befunden hatte, beziehungsweise an der Stelle ihrer Festung und der Wallburg Spas. Im siidostlichen Teil des ausgegrabenen Graberfeldes hat man ziemlich groBe Schuttablagerungen gefunden, unter denen sich auch Stiicke von Mortel mit Farbresten befanden. An dieser Stelle und in ihrer unmittelbaren Nahe wurden einige steinerne Fragmente mit vorromanischer F1echtwerkplastik gefunden , die zu Kircheneinrichtungsgegenstanden aus dem 9. oder 10. lh. gehorten. Die erwahnten Funde weisen mjt Sicherheit darauf hin, daB sich an eben dieser Stelle, wo man den Schutt gefunden hat, eine altkroatische Kirche befunden hatte, der die gefundenen Fragmente angehorten. Da sich von diesem Kirchenbau auch nicht der allergeringste Rest einer Mauer erhalten hat, kann man nur schlieBen, daB die Kirche bis zu den Grundmauern zerstort worden war, und daB ihre Steine als Baurnaterial fUr die Mauern der nahegelegenen Festung verwendet wurden. Der zwei te Teil dieser Abhandlung enthalt den Katalog der Grabarchitektur und ihres lnventars. Hier findet man kurz gefaBt die wichtigsten Angaben i.iber jedes cinzeIne Grab und die osteologischen Funde, sowie eine genaue Beschreibung der Grabbeigaben. Der dritte Teil behandelt die allgemeinen Merkmale des Graberfeldes. Wegen der ungi.instigen Bodenstruktur, sowie der Fundamente der Bauwerke, besitzt das Graberfeld nicht die Merkmale von Reihengrabern, Die Graber sind sehr verschiedenartig orientiert, und liegen in allen Richtungen; der groBte Teil der Graber war trotzdem richtjg orientiert, in der Richtung Osten-Westen (103 Graber) . In den meisten Grabern befand sich nur ein Verstorbener, manchmal jedoch auch mehrere. Die gefundenen osteologischen Reste bestatigen, daB das Graberfeld einer Siedlung zugehorig; mit einer gleich groBen Anzahl von bestatteten Mannern , Frauern und Kindern. Die Graber waren meistens aus behauener Rauhwacke , aus unregelmaBigen Steinplatten oder aus Bruchsteinen gebaut. Die behauene Rauhwacke ftir die Graber stammte wahrscheinlich von den eingesttirzten Bauwerken in der nahen Umgebung. Weiterhin analysiert der Verfasser die Grabfunde, meistens Schmuck, und zwar Ohrringe und Ringe. Indem er die typologischen Besonderheiten der einzelnen Gegenstande genau beurteilt, und sie mit analogem Material aus den tibrigen altkroatischen und anderen Graberfeldern vergleicht, kommt er zu Schltissen ber die Herkunft und\u27 Datierung der Funde. Mehr Aufmerksamkeit schenkt der Verfasser den Ohrringen vom Typ mit drei Beeren und zwar deshalb, weil in letzter Zeit versucht wurde, fast alle entwickelteren Varianten dieses Typs, die in Grabern auf dem Gebiet des altkroatisches Staates gefunden worden waren, von allen tibrigen Ohrringen die ebenfalls auf diesem Gebiet gefunden wurden, zu trennen. [ndem er auf das Problem der Datierung und Herkunft der Ohrringe mit drei Beeren eingeht, versuchte der Verfasser aufgrund einer Reihe sicherer Tatsachen und Hinweise darauf hinzuweisen, daB dieser Typ von Ohrringen, beziehungsweise die starker entwickelten Varianten , nicht von den tibrigen Typenvarianten altkroatischer Ohrringe isoliert werden dtirfen, die auf dem Gebiet des dalmatinischen Kroatiens gefunden worden waren , und die meistens in die Zeit vom Beginn des 9. bis zum Ende des 11. lahrhunderts datiert werden. Er machte ebenfalls darauf aufmerksam, daB samtliche Typen von altkroatischen Ohrringen, und so auch die entwickelteren Varianten von Ohrringen mit drei Beeren, die aus Grabern des Mutterlandes stammen, aus einheimischen Goldschmiedewerkstatten stammen , und daB sie als Ganzes eine altkroatische Kulturgruppe bilden. AuBer seinen Vermutungen tiber die Lage und GroBe der Ansiedlung, zu der das Graberfeld auf dem Berge Spas gehorte, ist der Verfasser im Ganzen der Meinun~, daB das Graberfeld auf Spas dem Zeitraum des frtihfeudalen kroatischen Staates angehorte, d. h., daB das Graberfeld sei t dem Anfang des 9. Jahrhunderts bestanden hatte und kontinuierlich bis zum Ende des ll. Jahrhunderts fortdauerte

‘Cytology-on-a-chip’ based sensors for monitoring of potentially malignant oral lesions

Despite significant advances in surgical procedures and treatment, long-term prognosis for patients with oral cancer remains poor, with survival rates among the lowest of major cancers. Better methods are desperately needed to identify potential malignancies early when treatments are more effective. Objective To develop robust classification models from cytology-on-a-chip measurements that mirror diagnostic performance of gold standard approach involving tissue biopsy. Materials and methods Measurements were recorded from 714 prospectively recruited patients with suspicious lesions across 6 diagnostic categories (each confirmed by tissue biopsy -histopathology) using a powerful new ‘cytology-on-a-chip’ approach capable of executing high content analysis at a single cell level. Over 200 cellular features related to biomarker expression, nuclear parameters and cellular morphology were recorded per cell. By cataloging an average of 2000 cells per patient, these efforts resulted in nearly 13 million indexed objects. Results Binary “low-risk”/“high-risk” models yielded AUC values of 0.88 and 0.84 for training and validation models, respectively, with an accompanying difference in sensitivity + specificity of 6.2%. In terms of accuracy, this model accurately predicted the correct diagnosis approximately 70% of the time, compared to the 69% initial agreement rate of the pool of expert pathologists. Key parameters identified in these models included cell circularity, Ki67 and EGFR expression, nuclear-cytoplasmic ratio, nuclear area, and cell area. Conclusions This chip-based approach yields objective data that can be leveraged for diagnosis and management of patients with PMOL as well as uncovering new molecular-level insights behind cytological differences across the OED spectrum

Plasma membrane dynamics and tetrameric organisation of ABCG2 transporters in mammalian cells revealed by single particle imaging techniques

ABCG2 is one of three human ATP binding cassette (ABC) transporters involved in the export from cells of a chemically and structurally diverse range of compounds. This multidrug efflux capability, together with a broad tissue distribution in the body, means that ABCG2 exerts a range of effects on normal physiology such as kidney urate transport, as well as contributing towards the pharmacokinetic profiles of many exogenous drugs. The primary sequence of ABCG2 contains only half the number of domains required for a functioning ABC transporter and so it must oligomerise in order to function, yet its oligomeric state in intact cell membranes remains uncharacterized. We have analysed ABCG2 in living cell membranes using a combination of fluorescence correlation spectroscopy, photon counting histogram analysis, and stepwise photobleaching to demonstrate a predominantly tetrameric structure for ABCG2 in the presence or absence of transport substrates. These results provide the essential basis for exploring pharmacological manipulation of oligomeric state as a strategy to modulate ABCG2 activity in future selective therapeutics